NZ527097A - Anti-cancer 2,3-dihydro-1H-pyrrolo[3,2-f]quinoline complexes of cobalt and chromium - Google Patents

Abstract

Disclosed herein are a class of heterocycles and their metal complexes, and the use of these compounds in the preparation of prodrugs or as prodrugs that may be activated under hypoxic conditions by enzymes or by therapeutic ionising radiation, in the treatment of cancer. The use of these heterocycles and the corresponding metal complexes in the preparation of medicaments and to compositions including the heterocycles or their metal complexes and to methods for preparing these compounds are also disclosed.

Description

527097 Anti-cancer 2,3-dihydro-lH- pyrrolo[3,2-f]quinoline complexes of Cobalt and Chromium.

The present invention relates to novel heterocycles and their metal complexes, and is particularly concerned with the use of these compounds in the preparation of prodrugs or as prodrugs that may be activated under hypoxic conditions by enzymes or by therapeutic ionising radiation, in the treatment of cancer. The • present invention also relates to the use of these novel heterocycles and their metal complexes in the preparation of a medicament and to methods for preparing these compounds.

BACKGROUND TO THE INVENTION Hypoxic regions occur widely in human tumours, and the cells in these regions are relatively resistant to ionising radiation. This leads to frequent recurrence of tumours after radiotherapy, due to the survival of these radioresistant cells. The use of oxygen-mimetic radiosensitizers has also been widely explored, but with mixed success. The existence of such hypoxic regions, restricted essentially to tumour tissue, has resulted in the development of bioreductive prodrugs (hypoxia-activated prodrugs; HAP) capable of being activated by enzymatic reduction only in these hypoxic regions. The majority of these prodrugs are activated to a transient one-electron intermediate in all cells, but this intermediate is re-oxidised by molecular oxygen in normal tissue, allowing activation to a toxic species to occur only in fully hypoxic cells.

The improved targeting ability of modern radiotherapy to deliver ionizing radiation only to the tumour field has suggested the possibility of using the reducing equivalents from this radiation, rather than cellular enzymes, to activate prodrugs (radiation-activated prodrugs; RAP). The activation of these prodrugs would thus be confined to hypoxic regions within the radiation field, providing a 2 double level of selectivity. Such, a mechanism of activation has other theoretical advantages over HAP [Wilson et al., Anticancer Drug Design, 13:663-685,1998]. These include: • Lack of collateral activation in partially hypoxic normal tissues (outside the radiation field).

• Use of the whole of the hypoxic tumour volume (including necrotic regions with no active reductases or reducing cofactors) to activate the prodrug.

• Avoidance of dependence on possibly varying enzyme levels, and degree of. 10 effectiveness as enzyme substrates.

While there have been many reports on HAP [for example reviews by Denny, Lancet Oncol. 2000,1,25-29; Stratford and Workman, Anti-Cancer Drug Design 1998,13, 519-528; Denny et al., Brit. J. Cancer, 1996, Suppl. 27,32-38], there has 15 been relatively few reports on RAP. An approach to using therapeutic ionizing radiation to activate a prodrug was reported [Nishimoto et al., J. Med. Chem. 1992, 35,2711; Mori et al; J. Org. Chem., 2000,65,4641-4647; Shibamoto et al., Jpn. J. Cancer Res., 2000,91,433-438; Shibamoto et al., Int. J. Rad. Oncol. Biol. Phys., 2001,49,407-413], employing radiolytic activation of a 5-fluorouracil (5-FU)-based 20 compounds, such as compound A. |l Radiolytic <y~u c acKvatif HN YV0H ^ HNL .NH H Y o B However, doses of radiation used during radiotherapy (typically 2 Gy/day) provide a total primary radical yield of only approximately 1.2 fimol/kg. Only about half of 25 this radical yield comprises reducing species capable of activating prodrugs by reduction. Thus the released effector 5-FU; illustrated as compound B above, is not 3 sufficiently potent to ensure clinically effective concentrations following therapeutic levels of irradiation.

The use of metal complexes of bidentate mustards, such as compound C 5 illustrated below, as RAP has also been reported [Denny et al., PCT NZ96/00085, 19 Aug 1996]. However, the released mustards, such as compound D illustrated below, are also unlikely to be sufficiently potent (IC50S around 1 |jM) to ensure clinically effective concentrations following therapeutic levels of irradiation.

It is therefore an object of the invention to provide heterocycles and their metal complexes either as prodrugs that are activated under hypoxic conditions by enzymes or other endogenous reducing agents or by therapeutic radiation, or at 15 least to provide the public with a useful choice.

+ D c SUMMARY OF THE INVENTION In a first aspect, the present invention provides a class of metal complexes 20 represented by Formula I I wherein: A is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci^alkyl, Ci^alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; B is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci^alkyl, Ci-6alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; D is selected from C or N; E is selected from a direct bond, OH or NR^, where each R1 independently represents H or a Ci.6alkyl optionally substituted with one or more hydroxy or amino groups, when D represents C; M is selected from Co111, Co11, Cr111 or Cr11; Z is selected from O, NR2, where R2 represents H or a Ci^alkyl optionally substituted with one or more hydroxy or amino groups, I ty S and S together represent formula V X o V wherein X is selected from a group including CEfe-halogen, CH20C0-(Ci-C6alkyl optionally substituted with one or more amino or hydroxy groups), diphosphate group or CH2OSO2R3, where R3 represents H or a Ci„6alkyl optionally substituted with one or more hydroxy or amino groups, or CH2OSO2NHR4 where R4 represents H or a C^alkyl optionally substituted with one or more hydroxy or amino groups; and R is selected from one of formulae VI or VII wherein each Ti, T2 and T3 is independently selected from H, OPO(OH)2, OR5, NR52 or NHCOR5, where each R5 independently represents H, a C^alkyl optionally substituted with one or more hydroxy or amino groups; or 0(CH2)nNR62, where each n is independently 1,2,3 or 4 and each R6 is 15 independently selected from H or a Ci-galkyl optionally substituted with one or more hydroxy or amino groups and • represents the point of attachment of R to Formula V defined above, and S3 is selected from H, cyano, phosphate, amino, C^allcy!, Ci-galkoxy, halogen, 20 C02[(Ci.6alkyl) wherein said alkyl is optionally substituted with amino, or hydroxy groups]; OR7, NR72, or CONHR7, where each R7 independently represents H, a Ci.6aUsyl optionally substituted with one or more hydroxy or amino groups; or S3 represents an optionally substituted 5 or 6 membered cyclic system optionally containing one or more hetroeroatoms fused to ring system A 25 defined above, wherein said substituents are selected from OH, cyano, phosphate, amino, Ci^alkyl, Ci-6alkoxy, and halogen groups, and 6 wherein ligands L!-L4 are each independently selected in combinations from anionic monodentate ligands, including CNF", SCN", halide, NO3"; bidentate ligands including MeCOCHJCOMe (Jacac; deprotonated in the complex), where J = H, Me, CI, SMe, S02Me, Me2NCS2"3 S(CH2)nS03H, S(CH2)„C02H, 5 S(CH2)„0P(0)(0H)2, CH2(CH2)nS03H, CH2(CH2)nC02H, CH2(CH2)n0P(0)(0H)2, S(CH2)nP(0)(0H)2 or CH2(CH2)nP(0)(0H)2, where n is from l-4;or tridentate ligands Vnia-VTIIc (= respectively TACH, TAME and TACNwhenRi-R3=H), Villa Vlllb Vlllc wherein each R1-R4 are independently selected from Hs Me, CH2(CH2)nSC>3H, CH2(CH2)„C02H, CH2(CH2)„P(0)(0H)2, CH2(CH2)n0P(0)(0H)2 or CH2(CH2)nNR82> where each n is independently 1,2,3 or 4 and each R8 15 independently represents H, or a Ci^alkyl optionally substituted with one or more hydroxy or amino groups or LJ-L4 can also be selected from any one of the tetradentate ligands IX-XVII, or any two of the bidentate ligands XVIII, or any combination of the bidentate 20. ligands XVIII together with any of the monodentate ligands L'-L4 defined above; wherein in formulae IX-XVEH, R1 to R8 each independently represent H, Me, 5 CH2(CH2)nS03H, CH2(CH2)„C02H, CH2(CH2)nP(0)(OH)or CH2(CH2)n0P(0)(0H)2 or CH2(CH2)nNMe25 where each n is independently 1,2, 3 or 4; each Z^Z4 is independently selected from -(CH2)2-, -(CH2)3-, -CH2OCH2- or -CH2N(R9)CH2-; where R9 represents H, a Ci.6alkyl optionally substituted with one 10 or more hydroxy or amino groups and each Y' is independently selected from H, halogen, SC^Me, 0(Cj-C6alkyl), NR102j where each R10 is independently selected from H or a Ci-ealkyl optionally substituted with one or more hydroxy or amino groups, or Q1(GH2)nQ2, wherein Q1 is selected from -0-, -CH2-, -NH-, -CONH-, -C02- or -S02-, and Q2 is 15 selected from -C02H, -S03H, -OP(0)(OH)2 or -NR1 \ where each R11 is independently selected from H or a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups; and wherein the overall charge on the complex is neutral, positive or negative and wherein in the case of a non-neutral complex Foounteroharge is selected from a range 20 of physiologically acceptable-counterions, including halide", N03", NH/ or Na+; and 8 wherein q is the required number to neutralise the overall charge on the complex; and including any enantiomeric or diastereomeric form, and any physiologically salt derivative thereof.

Preferably, the rings A and B of a compound of Formula I as defined above together represent an 8-substituted quinoline system.

In a further aspect the present invention provides a method of providing cancer treatment, which includes the steps of 10 (a) administering to a patient in need of such therapy an effective amount of a compound of Formula I as defined above, and (b) activating the compound of Formula I under hypoxic conditions via reduction, either enzymatically or by non-enzymatic endogenous reducing agents, or by ionizing radiation, wherein said activation releases a sufficient amount of an effector firom said effective amount of the compound of Formula I.

In a further aspect the present invention further provides a composition comprising as an active agent a compound of Formula I as defined above and a 20 pharmaceutical^ acceptable excipient, adjuvant or carrier.

In a further aspect the present invention provides the vise, in the manufacture of a medicament, of an effective amount of a compound of Formula I for use in treating a subject in need of cancer treatment.

In another aspect, the present invention provides a class of metal complexes represented by Formula la 9 charge qpcountercharge (la) wherein: A is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Cj^alkyl, Ci-galkoxy, halogen, hydroxy, phosphate, cyano or amino groups; B is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci^alkyl, 10 Ci-6alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; D is selected firom C or N; E is selected firom a 5 or 6 membered ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci-6alkyl, Ci-galkoxy, halogen, hydroxy, phosphate, cyano or amino groups; 15 M is selected from Com, Co11, Cr111 or Cr11; Z represents NH2 or NHMe; Q represents H, C^alkyl or when Q represents Q will become a ligand for M and replace one of ligands L!-L4 defined below, S1 and S2 together represent formula V X V wherein X is selected firom a group including CHh-halogen, CEbOCO-CCi-Cealkyl optionally substituted with one or more amino or hydroxy groups), CEfe-phosphate group or CH2OSO2R3 where R3 represents H or a Ci^alkyl optionally 5 substituted with one or more hydroxy or amino groups, or CH2OSO2NHR4 where R4 represents H or a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups; and R is selected firom one of formulae VI or VII wherein each Ti, T2 and T3 is independently selected firom H, OPO(OH)2, OR2, m22 where each R2 independently represents H, a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups or 0(CH2)nNR32, where each n is 15 independently 1,2, 3 or 4, and each R3 is independently selected from H or a Ci-gallcyl optionally substituted with one or more hydroxy or amino groups and • represents the point of attachment of R to Formula V defined above, and S3 is selected from H, cyano, phosphate, amino, Ci^alkyl, Cj^alkoxy, halogen, C02(Ci-6alkyl) wherein said alkyl is optionally substituted with amino, or halogen 20 groups, OR4, NR42, CONHR4, where each R4 independently represents H, a Ci-galkyl optionally substituted with one or more hydroxy or amino groups; or S3 represents an optionally substituted 4-8 membered cyclic system optionally containing one or more heteroatoms fused to ring system A defined above, wherein said substituents are selected firom OH, cyano, phosphate, amino, 25 Ci.6alkyl, Ci^allcoxy, halogen groups, and VII '3 VI 11 wherein ligands Ll-L4 are each independently selected in combinations firom anionic monodentate ligands, including CN", SCN", halide, NO3"; bidentate ligands including MeCOCHJCOMe (Jacac), where J = H, Me, CI, SMe, S02Me, S(CH2)„S03H, S(CH2)nC02H, S(CH2)n0P(0)(0H)2, CH2(CH2)nS03H, CH2(CH2)nC02H or CH2(CH2)n0P(0)(0H)2s where each n is independently 1, 2, 3 or 4; or tridentate ligands Vnia-YIIIc (= respectively TACH, TAME and TACN when R1-R3HS), wherein Ri-R4 are each independently selected from H, Me, CH2(CH2)nS03H, CH2(CH2)aC02H or CH2(CH2)n0P(0)(0H)2 or CH2(CH2)nNR52, where each n is independently 1,2,3 or 4 and each R5 independently represents H, or a Ci.6alkyl optionally substituted with one or more hydroxy or amino groups or L'-L4 can also be selected firom any one of the tetradentate ligands IX-XVII, or any two of the bidentate ligands XVIH, or any combination of the bidentate ligands XVm together with any of the monodentate ligands L^L4 defined above; r2hn Vlllb Villa Vlllc 12 wherein in formulae IX-XVUI, R1 to R8 each independently represent H, Me, 5 CH2(CH2)„S03H, CH2(CH2)„C02H or CH2(CH2)n0P(0)(0H)2 or CH2(CH2)nNMe2, where each n is independently 1,2,3 or 4; each Zr-Z4' is independently selected firom -(CH2)2-, -CH2OCH2- or - CH2N(R6)CH2-; where R6 represents H, a Ci^alkyl optionally substituted with one or more hydroxy or amino groups and 10 each Y' is independently selected from H, halogen, SC^Me, (XCi-Cgallcyl), NR72, where each R7 is independently selected firom H or a Ci^alkyl optionally substituted with one or more hydroxy or amino groups, or Q1(CH2)nQ2, wherein Q1 is selected firom -0-, -CH2-, -NH-, -CONH-, -C02- or -S02:, and Q2 is selected from -C02H, -SO3H, -0P(0)(0H)2 or ~NR82 where each R8 is 15 independently selected firom H or a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups; and wherein the overall charge on the complex is neutral, positive or negative and wherein in the case of a non-neutral complex Fcounterchar8e is selected firom a range of physiologically aceeptable-counterions, including halide", NO3", NH41" or Na+; 20 and WO 02/059122 PCT/NZ02/00005 13 wherein q is the required number to neutralise the overall charge on the complex, and including any enantiomeric or diastereomeric form, and any physiologically salt derivative thereof.

Preferably, the rings A and B of a compound of Formula la as defined above together represent an 8-substituted quinoline system.

In a further aspect the present invention provides a method of providing cancer treatment, which includes the steps of 10 (c) administering to a patient in need of such therapy an effective amount of a compound of Formula la as defined above, and (d) activating the compound of Formula la under hypoxic conditions via reduction, either emzymatically or by non-enzymatic endogenous reducing agents or ionizing radiation, wherein said activation releases a sufficient amount of an effector, from said effective amount of the compound of Formula la, which is of sufficient potency to kill cancer cells.

In a further aspect the present invention further provides a composition 20 comprising as an active agent a compound of Formula la as defined above and a pharmaceutically acceptable excipient, adjuvant or carrier.

In a further aspect the present invention provides the use, in the manufacture of a medicament, of an effective amount of a compound of Formula la for use in 25 treating a subject in need of cancer treatment.

In another aspect, the present invention provides a class of heterocycles of Formula XIX. 14 S Z XIX wherein A is selected from a 5 or 6 membered aromatic ring system optionally containing 5 one or more additional heteroatoms and optionally substituted with one or more" Ci.6alkyl, Ci.6allcoxy, halogen, hydroxy, phosphate, cyano or amino groups; B is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci^alkyl, Ci^alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; 10 Z is selected from OH or NR.12, where R1 separately represent H or Ci-Qalkyl optionally substituted with one or more amino, hydroxy, a halogen or cyano groups; S1 and S2 together represent formula V wherein X is selected from a group including CH2-halogen, CH2OCO'(CrCsalkyl optionally substituted with one or more amino or hydroxy groups), CH2- phosphate group or CH2OSO2R3 where R3 represents H or a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups, or CH2OSO2NHR4 where R4 represents H or a Cj^alkyl optionally substituted with one or more hydroxy or amino groups; and R is selected from one of formulae VI or VII X O V VII T3 VI wherein each Ti, T2 and T3 is independently selected from H, OPO(OH)2, OR5, NR52 where each R5 independently represents H, a Cj-6alkyl optionally substituted 5 with one or more hydroxy or amino groups or 0(CH2)nNR62, where each n is independently 1,2,3 or 4 and each R6 is independently selected from H or a Ci-gallcyl optionally substituted with one or more hydroxy or amino groups; • represents the point of attachment to Formula V defined above; S3 is selected from H, cyano, phosphate, amino, Ci.6alkyl, Cj^allcoxy, halogen, 10 C02[(Ci-saIkyl) wherein said alkyl is optionally substituted with amino, or hydroxy groups], OR7, NR72, CONHR7 where each R7 independently represents H, a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups; or S3 represents an optionally substituted 4-8 membered cyclic system optionally containing one or more hetoeroatoms fused to ring system A defined above, 15 wherein said substituents are selected from OH, cyano, phosphate, amino, Ci.6alkyl, Ci^alkoxy, and halogen groups, and including any enantiomeric or diastereomeric form, and any physiologically salt derivative thereof. with the proviso that when Z, A, B, X, S1, S2 and S3 together represent OH R does not represent one of the following 16 Preferably, the rings A and B of a compound of Formula XEX as defined above together represent an 8-substituted quinoline system.

In a further aspect the present invention provides a method of providing cancer treatment, which includes the step of administering to a patient in need of such therapy an effective amount of a compound of Formula XIX wherein A is selected from a 5 or 6 membered aromatic ring system optionally containing one or more additional heteroatoms and optionally substituted with one or more 15 Ci-galkyl, Ci.6alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; B is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci^alkyl, Ci-6alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; Z is selected from OH or NR1^ where R1 separately represent H or Ci-Cgalkyl 20 optionally substituted with one or more amino, hydroxy, a halogen or cyano groups; S z XIX 17 S1 and S2 together represent formula V X )"Y-R O V wherein X is selected from a leaving group including CH2-halogen, CH2-5 phosphate group, CH2OCO R2, where R2 represents Ci-C6alkyl optionally substituted with one or more amino or hydroxy groups; CH2OSO2R3 where R3 represents H or a Chalky! optionally substituted with one or more hydroxy or -amino groups, or CH2OSO2NHR4 where R4 represents H or a Ci-galkyl optionally substituted with one or more hydrogen or amino groups; and 10 R is selected from one of formulae VI or VII wherein each Ti, T2 and T3 is independently selected from H, OPO(OH)2, ORs, 15 NR52 where each Rs independently represents H, a Ci.6alkyl optionally substituted with one or more hydroxy or amino groups or 0(CH2)nNR62, where each n is independently 1,2,3 or 4 and each R6 is independently selected from H or a C;_6 alkyl optionally substituted with one or more hydroxy or amino groups; * represents the point of attachment to Formula V defined above; S3 is selected from H, cyano, phosphate, amino, Ci.6alkyl, Ci.6alkoxy, halogen, C02[(Ci-6allcyl) wherein said alkyl is optionally substituted with amino or hydroxy groups], OR7, NR72, CONHR7 where each R7 independently represents H, a Ci-galkyl optionally substituted with one or more hydroxy or amino groups; or S3 represents an optionally substituted 4-8 membered cyclic system optionally 25 containing one or more hetoeroatoms fused to ring system A defined above, wherein said substituents are selected from OH, cyano, phosphate, amino, 18 Ci-6alkyl, Ci-galkoxy, and halogen group, and including any enantiomeric or diastereomeric form, and any physiologically salt derivative thereof.

In a further aspect the present invention provides a composition comprising as an active agent a compound of Formula XIX S3 Z XEX wherein A is selected firom a 5 or 6 membered aromatic ring system optionally containing 10 one or more additional heteroatoms and optionally substituted with one or more Ci-6alkyl, Ci-galkoxy, halogen, hydroxy, phosphate, cyano or amino groups; B is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci-galkyl, Ci-galkoxy, halogen, hydroxy, phosphate, cyano or amino groups; 15 Z is selected from O or NR1, where R1 represents H or Ci-Cgalkyl optionally substituted with one or more amino, hydroxy, a halogen or cyano groups; S1 and S2 together represent formula V \ O V wherein X is selected from a leaving group including CBb-halogen, CH2-20 phosphate group, CH2OCOR2, where R2 represents Ci-Cealkyl optionally substituted with one or more amino or hydroxy groups; CH2OSO2R3 where R3 represents H or a Ci-6alkyl optionally substituted with, one or more hydroxy or amino groups, or CH2OSO2NHR4 where R4 represents H or a Ci^alkyl optionally substituted with one or more hydroxy or amino groups; and 19 R is selected firom one of formulae VI or VII wherein each Ti, T2 and T3 is independently selected from H, OPO(OH)2, OR5, NR52 where each R5 independently represents H, a Chalky! optionally substituted with one or more hydroxy or amino groups or 0(CH2)nNR62, where each n is independently 1,2,3 or 4 and each R6 is independently selected firom H or a Ci-galkyi optionally substituted with one or more hydroxy or amino groups; 10 • represents the point of attachment to Formula V defined above; S3 is selected firom H, OH, cyano, phosphate, amino, Chalky!, Ci^alkoxy, halogen, C02[(Ci-galkyl) wherein said alkyl is optionally substituted with amino, or hydroxy groups], OR7, NR7, CONHR7 where each R7 independently represents H, a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups; 15 or S3 represents an optionally substituted 4-8 membered cyclic system optionally containing one or more hetoeroatoms fused to ring system A defined above, wherein said substituents are selected firom OH, cyano, phosphate, amino, Ci^alkyl, Ci^alkoxy, and halogen groups; and including any enantiomeric or diastereomeric form, and any physiologically salt derivative thereof, and 20 a pharmaceutically acceptable excipient, adjuvant or carrier.

In a further aspect the present invention provides the use, in the manufacture of a medicament, of an effective amount of formula XIX S Z xrx wherein A is selected from a 5 or 6 membered ring system optionally containing one or 5 more additional heteroatoms and optionally substituted with one or more Ci-ealkyl, Ci^alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; B is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci-gallcyl, Ci-galkoxy, halogen, hydroxy, phosphate, cyano or amino groups; 10 Z is selected from OH or NR^, where each R1 independently represents H or Ci-Cgalkyl optionally substituted with one or more amino, hydroxy, a halogen or cyano groups; S1 and S2 together represent formula V. wherein X is selected from a leaving group including CH2-halogen, CH2- phosphate group, CH2OCOR2, where each R2 independently represents Ci-Cgalkyl optionally substituted with one or more amino or hydroxy groups; CH2OSO2R3 where R3 represents H or a Chalky! optionally substituted with one or more hydroxy or amino groups, or CH2OSO2NHR5 where R5 represents H or a Cnjalkyl 20 optionally substituted with one or more hydroxy or amino groups; and X o V R is selected from one of formulae VI or VII 21 wherein each Ti, T2 and T3 is independently selected from H, OPO(OH)2, OR5, NR52 where each R5 independently presents H, a Ci-galkyl optionally substituted 5 with one or more hydroxy or amino groups or 0(CH2)nNR62, where each n is independently 1,2,3 or 4, and each R6 is independently selected from H or a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups; • represents the point of attachment to Formula XIX defined above; S3 is selected from H, OH, cyano, phosphate, amino, Chalky!, Ci-6alkoxy, 10 halogen, C02[(Ci.6allcyl) wherein said alkyl is optionally substituted with amino, or hydroxy groups], OR7, NR7, CONHR7 where each R7 independently represents H, a Ci^alkyl optionally substituted with one or more hydroxy or amino groups; or S3 represents an optionally substituted 4-8 membered cyclic system optionally containing one or more hetoeroatoms fused to ring system A defined above, 15 wherein said substituents are selected from OH, cyano, phosphate, amino, Ci-galkyl, Ci-galkoxy, and halogen groups, for use in treating a subject in need of cancer treatment, and including any enantiomeric or diastereomeric form, and any physiologically salt derivative thereof.

It is to be recognised that the compounds of the invention defined above can exist in different enantiomeric and/or diastereomeric forms. In such cases it is to be understood that formulae I, la and XIX can represent any possible enantiomeric or diastereomeric form, or any mixtures of such forms, and also any physiologically functional salt derivatives thereof.

In a final aspect, the present invention provides methods of preparing compounds of the general formulae I, la and XIX defined above. Such methods are described below. 22 It is to be understood that the terms Cms alkyl and Ci-6 alkoxy as used throughout the specification are to be taken as including both the straight and branched forms of such groups.

DESCRIPTION OF THE DRAWINGS While the invention is broadly defined above, it will be appreciated by those skilled in the art that further aspects of the invention will become apparent with _ 10 reference to the following Figure and Examples, given by way of example only, wherein Figure 1 shows graphically the release of a cytotoxic effector 18a (SN 26800) from a compound of Formula I Ml (SN 27892) when irradiated in formate buffer, 15 pH 7.0 under hypoxic conditions.

Figure 2 shows graphically the hypoxic selectivity of metal complex Ml in HT29 cultures.

DETAILED DESCRIPTION OF THE INVENTION As defined above, this invention provides novel heterocycles and their metal complexes, and is particularly concerned with the use of these compounds, as pro drugs activated under hypoxic conditions by enzymes or by therapeutic ionising 25 radiation, in the treatment of cancer.

In order to ensure that the complexes (pro drugs) of Formula I and la and the heterocycles (cytotoxins or effectors) of Formula XIX of the present invention are clinically effective, the complexes and heterocycles would preferably have the 30 following properties - high chemical stability of the +M metal oxidation states 23 - minimal toxicity as a prodrug prior to reductive activation by enzymes or radiation - upon activation the prodrug releases a potent cytotoxic or effector unit Examples of pro drug complexes or heterocyclic compounds that fulfil these general criteria include the compounds of Formula I, la or XIX as defined above. These compounds can be prepared by the following schemes and processes as described below by way of example only.

Preparation Example 1 A: Synthesis of 5-hvdroxv-2.3-dihvdropvrrolor3.2-flauinolines In general, 5-hydroxy-2,3-dihydropyrrolo[3,2-fjquinolines of formula (XIX; Z=0) can be made from the precursor 14, that in turn can be prepared by the 15 method outlined in Scheme 1. Conversion of the known [Curd et al., J. Chem. Soc., 1947, 69,1613] 1 by the Slcraup reaction gives 2 in 80% yield, using an improved procedure [Battersby et al., J. Chem. Soc. Perkin Trans. I, 1979,2250]. Conversion of methyl to benzyl (2-»3-*4) (to allow more ready removal at the end of the synthesis), followed by reduction of 4 with Fe/AcOH gives 5. This can 20 beBOC-protected to give 6, which can be iodinated with NIS/MeCN to give 7a or brominated (NBS/MeCN) to give 7b. Alkylation of 7a/7b with 3-bromo-l,l-dimethoxypropane gives 8a/8b, which can be deprotected (TsOH) to 9a/9b, then converted to the vinyl acetates. lOa/lOb (AC2O, DMAP, THF, reflux). These undergo radical cyclization (Bu3SnH/AIBN) to give 11, which can be deprotected 25 (CS2CO3) to give 12. This can be converted either directly (PI13P, CCI4) or via mesylate 13 (MsCl, Et3N; then LiCl, DMF) to the desired racemic pyrroloquinoline 14. 24 Scheme 1 H2N no2 f 1 OMe 11: R = OAc ^l&RsOH —I =*=- 13: R « OMs <-»-14: R = CI -*-!• &2:R = (Jle 3: R = H 4: R = Bn OBn r— 5: R = NH, U^6: R » NHBOC NHBOC OBn 7a: R = I 7b: R = Br ■OAc NBOC OBn 10a: R = I 10b: R = Br .OMe NBOC 8a: R = I 8b: R = Br An alternative and shorter route from 7a/7b to 14 is shown in Scheme 2. N-Alkylation of 7a/7b with 1,3-dichloropropene, and radical cyclization of the resulting vinyl chlorides 15a/15b gives 14 in high yield.

Scheme 2 7a or 7b 14 OBn 15a: R = I 15b: R = Br The benzyl group of 14 can be removed by hydrogenolysis (Scheme 3), and the resulting phenol 16 can be N-deprotected and coupled with appropriate side chains R (formula XIX). An alternative route is by N-deprotection/coupling, followed by removal of the benzyl group (14-»17-»18), either by hydrogenolysis or by acid treatment.

Scheme 3 e: R = .OMe f:R = OMe Preparation Example 2 Synthesis of 5-amino-7-3-dibvdropvm)lor3.2-flauinolines 5 In general, 5-amino-2,3-dihydropyrrolo[3,2-f]quinolines (XIX, Z represents NH2) can be prepared firom the precursor 25, which can be synthesized by the method outlined in Scheme 4. Hie quinoline acid 19 [Jung et al., Eur. Pat. Appln. EP 581500 (1994); Chem Abstr, 1994,122,205125], prepared by a Skraup reaction on ethyl 4-amino-3-mtrobenzoate, is converted with DPPA/t-BuOH/Et3N to the 10 quinoline 20. Nitro group reduction gives amine 21, which is converted to the phthaloyl derivative 22, and then brominated (NBS/MeCN) to give 23. N-Allcylation of this with 1,3-dichloropropene, followed by radical cyclisation of the resulting chloro intermediate 24 with BusSnH/AIBN, gives the tricyclic pyrroloquinolinone 25. As shown in Scheme 4, NBOC deprotection of 25 15 followed by EDCI coupling with acids gives the compounds of formula 26 (illustrated for the example where R=5,6,7-trimethoxyindol-2-yl). Finally, deblocking of compound 26 by hydrazinolysis gives compounds of foraiula XIX, WO 02/059122 PCT/NZ02/00005 26 where Z represents NH2 (illustrated for the example where R=5,6,7- trimethoxyindol-2-yl; 27).

Scheme 4 NHBOC N02 R C 20: R=NH!30C ^ 22 R=t&hai NPhthal 23: R=NHBOC 24: R=N(BOC)CH=CHCI NBOC r- 26: X=NPhthal I— 27: X=NH2 OMe R- jfO~0Me /^N OMe Preparation Example 3 Synthesis of 5-(2-aminoethvlamino')benzrel indoles These can be prepared from the appropriate 5-amino compounds by condensation with the BOC-protected aminoacetaldehye, followed by reduction with sodium cyanoborohydride or other suitable reductants, and deprotection of the BOC group. Scheme 5 shows the synthesis of the representative compound 29 from the known [Atwell et al., J. Org. Chem. 1998,63, 9414-9420] 5-amino compound 28. It will be appreciated that this synthesis can also be applied to the preparation of the analogous derivative from the 5-aminoaza compound 27.

Scheme 5 BOCNI-r OMe RHN" r~ R = BO J U-R = H: Preparation Example 4 27 Synthesis of ancillary ligands As an example of the synthesis of new ancillary cyclen-type tetradentate ligands, reaction of perhydro-3,6,9,12-tetraazacyclopenteno[l,3-^g]acenaphthylene (30) [Weisman et al., Tetrahedron Lett., 21,1980, 335] with 1,3-propanesultone gives the bis-quaternary salt (31), which is treated with hydrazine monohydrate to give the bis(propanesulfonic acid) (32) (Scheme 6). It will be appreciated that similar reaction of 30 with other alkylating reagents will give other analogues, such as those represented as compounds 33 to 36 in Scheme 6.

Scheme 6 OO u >i Preparation Example 5 Synthesis of metal complexes As an example of the synthesis of metal complexes of Formula I defined above, using a tetradentate ancillary ligand, reaction of complex 39 bearing labile triflate ligands with 18a gives the Co111 complex Ml, as illustrated in Scheme 7. 28 As an example of the synthesis of metal complexes of Formula la defined above, using bidentate ancillary ligands, reaction of 18a with [Cr(acac)2(H20)2]CIO4] -2H20 in dry CH3CN gives the desired Cr(acac)2-18a complex M4. This reaction pathway is represented in Scheme 8. Similar reaction of 29 gives the corresponding Cr(acac)2-29 complex M6.

CrIU complexes with other tetradentate macrocycles may be prepared by a similar synthetic route to that employed in the example above using Co111, in that the key 29 intermediate for both is a reactive bis[triflato] complex (or a solvent species in solution). The use of nitro complexes as precursors to triflato complexes is unlikely for Cr111, as nitro is a poor ligand on Cr111. Instead, chloro complexes serve the purpose well. In the strongly acidic triflic acid, protonation of coordinated CI" 5 is significant and leads to labilization and ligand loss, made irreversible by removal of the gaseous HC1 co-product.

As an example of metal complexes with tridentate ligands, reaction of the triamine TACN with Na3[Co(N02)fi] gives the complex 10 [Co(TACN)(H20)3].(OTf)3 (Scheme 9). Reaction of this with the model quinoline 8-hydroxyquinline (8-HQ) gives complex M7.

EXAMPLES OF THE INVENTION The following examples of metal complexes M1-M9 in Table 1 are representative of the complexes of the invention and can be prepared by the detailed processes of the invention described after the table. 8-HQ OH + Table 1. Structures and physical properties of metal complexes No Metal toxic ligand Ancillary ligands Analyses Ml Com 18a Cyclen (IX; Z*-Z4 = -(CH2)2-,R1'-R4' = H) C32H4iN7d:,ClCo05 [M-2C104-H]+ Calc: 697.21897 Fd: 697.21327 M2 ■ Com 18c Cyclen (IX; Zx-Z4 = -(CH2)2-,R1'-R4,= H) C33H45J5ClCoN703 [M-20Tf]+ Calc: 681.26044. Fd: 681.26064 M3 Com 18b Cyclen (IX; Z!-Z4 = -(CH2)2-, R1-R4'=H) C33H443SClCoN803 [M-20Tf]+ . Calc: 694.25569. Fd: 694.25305 M4 Crlu 18a (Acac)2 (MeCOCH2COMe)2 C34H36N3jSClCr09 [M+H]+ Calc: 717.15452 Fd: 717.15198 M5 CoJ11 18a (Me2dithiocarbamato)2 (Me2NSC2)2 M6 Crin 29 (Acac)2 (MeCOCH2COMe)2 C37H43N4J5ClCr08 [M-C104]+ Calc: 758.21745 Fd: 758.21834 M7 Co111 8-HQ TACN (VIHc: RJ-RJ = H)) 1 Example A. Preparation of l-fchloromethvlV5-hvdroxv-3-r(5.6.7-trimethoxvindol-2-vl)carbonvl]-2.3-dihvdro-li?-pvrrolor3.2-flquinoline (18a^ and analogues 18b-18f by the methods of Schemes 1-3. 8-Hydroxy-6-nitroquinoline hydrobromide (3). A solution of 8-methoxy-6-nitroquinoline (2) [prepared from 2-methoxy-4-nitroaniline 1 by the method of Battersby et al., J. Chem., Soc. Perlcin Trans. 1,1979,2550] (50.0 g, 0.245 mol) in 48% aqueous HBr (0.205 L, 1.22 mol) was stirred at reflux for 65 h. The mixture was cooled in ice and the precipitate was removed by filtration and dried in a 10 desiccator to give 3 as the hydrobromide salt (58.0 g, 87%): subl. 140 °C, mp >230 °C; NMR (DMSO) 5 10.69 (br s, 2 H), 9.20 (dd, J= 4.9,1.5 Hz, 1 H), 31 9.11 (dd, 8.5,1.5 Hz, 1 H), 8.64 (d, /= 2.4 Hz, 1 H), 8.05 (dd, J= 8.5,4.9 Hz, 1 H), 7.90 (d, J= 2.4 Hz, 1 H); 13C NMR (DMSO) 5 152.0,149.4,146.4,144.3, 135.4, 128.3,124.1,114.5,106.5. Anal. Calcd for C9H6N2O3.HB1:: C, 40.01; H, 2.61; N, 10.37. Found: C, 40.44; H, 2.17; N, 10.83. 8-Benzyloxy-6-mtroquinoIine (4). A mixture of 3 (58.0 g, 0.214 mol), DMF (400 mL), K2CO3 (103.5 g, 0.75 mmol), andNal (1.60 g,"10.7 mmol) was stirred at room temperature, while benzyl bromide (25.4 mL, 0.214 mmol) was added in four portions at half hourly intervals. A total of 9 h after the first addition, the 10 mixture was poured onto ice (1.5 kg) and the precipitate was removed by filtration, washed with water, and dried. The crude material was dissolved in CH2CI2 and the solution was filtered through alumina to give 4 (59.55 g, 99%): mp (EtOH) 152-153 °C; ^NMR^DCls) 5 9.13 (dd,J= 4.2,1.8 Hz, 1 H), 8.35 (d, J= 2.3 Hz, 1 H), 8.29 (dd, /= 8.4,1.8 Hz, 1 H), 7.83 (d, J- 2.3 Hz, 1 H), 7.59 15 (dd, J= 8.4,4.2 Hz, 1 H), 7.56 (d, 7.6 Hz, 2 H), 7.40 (dd, J= 7.6, 7.2 Hz, 2 H), 7.33 (t, J= 7.2 Hz, 1 H), 5.50 (s, 2 H); 13C NMR. (CDCI3) S 155.4, 152.5, 145.6, 142.6,137.9,135.4,128.8, 128.4, 127.8,127.5,123.3, 116.3,103.1, 71.4. Anal. Calcd. for Ci6Hi2N203: C, 68.57; H, 4.32; N, 9.99. Found: C, 68.51; H, 4.29; N, 10.04. 6-Amino-8-benzyloxyquinoIine (5). Iron dust (16.0 g, 0.285 mol) was added to a solution of 4 (8.00 g, 28.5 mmol) and AcOH (16 mL, 0.285 mol) in EtOH-water (5:1,240 mL) at reflux. After 10 min, the mixture was carefully poured into saturated aqueous NaHCCb (300 mL). The mixture was filtered through Celite 25 and the filter cake was washed with water (100 mL), EtOH (3 x 50 mL), and CH2C12 (3 x 100 mL). The combined filtrates were diluted with water (300 mL) and the aqueous layer was separated and extracted with CH2CI2 (2 x 50 mL). The combined extracts were washed with water, dried (NazSO^i), and evaporated to give 5 (7.13 g, 100%) as a tan solid: mp 183-185 °C; *H NMR (CDCI3) 8 8.66 30 (dd, /= 4.2,1.6,Hz, 1 H), 7.84 (dd, J= 8.3, 1.6 Hz, 1 H), 7.48 (dd, J~ 8.1,1.7 Hz, 2 H), 7.23-7.39 (m, 3 H), 7.28 (dd, /= 8.3,4.2 Hz, 1 H), 6.51,6.48 (2 x d, J WO 02/059122 PCT/NZ02/00005 32 = 2.3 Hz, 1 H each), 5.36 (s, 2 H), 3.85 (br s, 2 H); 13C NMR (CDCI3) 8 155.2, 155.7,144.8,136.8,135.9,133.5,130.8,128.6,127.8,127.0,122.0,102.6,100.0, 70.6. Anal. Calcd. for Ci6H14N20: C, 76.78; H, 5.64; N, 11.19. Found C, 76.54; H, 5.61; N, 11.15. 8-Benzyloxy-6-(fe/t-butyloxycarbonylamino)quinoline (6). A mixture of 5 (7.63 g, 30.5 mmol), BOC2O (8.65 g, 39.6 mmol) and dioxane (70 mL) was stirred at reflux for 2 h. Further BOC2O (0.86 g, 4.0 mmol) was added and the mixture was heated at reflux for another 1 h. The dioxane was evaporated, the 10 remaining oil was triturated with pentane, and the resulting solid was removed by filtration, dissolved in CH2CI2 and filtered through alumina to give 6 (10.42 g, 98%) as a cream solid: mp 180-181 °C; NMR (CDC13) 8 8.77 (dd, J= 4.2,1.6 Hz, 1 H), 7.98 (dd, J= 8.3,1.6 Hz, 1 H), 7.55 (d, J= 2.1 Hz, 1 H), 7.41 (dd, J= 7.4,2.2 Hz, 2 H), 7.34 (dd, J= 8.3,4.2 Hz, 1 H), 7.20-7.29 (m, 3 H), 7.02 (d, J= 15 2.1 Hz, 1 H), 5.28 (s, 2 H), 1.49 (s, 9 H); 13C NMR (CDCl3) S 154.6, 152.7,147.4, 137.2,136.8, 136.3, 135.2,129.9,128.4, 127.7,127.2, 122.0,105.8,103.5, 80.6, 70.6,28.2. Anal. Calcd for C21H22N2O3: C, 71.98; H, 6.33; N, 7.99. Found C, 71.80; H, 6.31; N, 7.98. 8-Benzyloxy-6-(ter^-butyloxycarbonylamino)-5-iodoquinoline (7a). A mixture of 6 (1.04 g, 3.0 mmol), NTS (0.70 g, 3.1 mmol) and CH3CN (10 mL) was stirred at reflux for 30 min. Further NIS (40 mg, 0.18 mmol) was added and the mixture stirred at reflux for a further 30 min. The CH3CN was evaporated and the residue was taken up in EtOAc (30 mL) and washed with a solution ofNa2S2C>5 and 25 Na2CC>3 in water (x 3). The aqueous washes were back extracted with EtOAc (x 2). The combined organic extracts were washed with water, dried (brine, MgS04), filtered through silica gel, and evaporated to give 7a (1.33 g, 93%), which crystallized from hexane as tan needles: mp 118-119 °C; *H NMR (CDCI3) 8 8.79 (dd, J= 4.2,1.4 Hz, 1 H), 8.32 (dd, J= 8.6,1.4 Hz, 1 H), 8.29 (s, 1 H), 7.59 (dd, / 30 = 8.0,1.7 Hz, 2 H), 7.43 (dd, /= 8.6,4.2 Hz, 1 H), 7.25-7.39 (m, 3 H), 7.24 (br s, 1 H), 5.43 (s, 2 H), 1.57 (s, 9 H); 13C NMR (CDC13) 8 155.2,152.4,148.1,139.5, 33 138.9,138.3, 136.2,130:7,128.5,128.0,123.4,103.9,81.5,78.1,71.0,28.3.

Anal. Calcd for C21H21IN2O3: C, 52.96; H, 4.44; N, 5.88. Found C, 53.18; H, 4.39; N, 5.95. 8-Benzyloxy-6-[jV-(fert-butyIoxycarbonyl)-iV-(3,3-dunethoxypropyl)ammo]-5-iodoquinoline (8a). NaH (60% in oil, 92 mg, 2.3 mmol) under nitrogen was washed with pentane (2x2 mL), cooled (ice-water) and treated with a solution of 7a (1.00 g, 2.10 mmol) in DMF (10 mL) over 5 min. The mixture was allowed to warm to room temperature and stir for 30 min, over which time it became bright 10 yellow and effervescence ceased. A solution of 3 -bromo-1,1 -dimethoxypropane (0.69 g, 3.77 mmol) in DMF (0.5 mL) was added and the mixture was stirred at room temperature for 22 h. The mixture was poured into pH 7.4 phosphate buffer (50 mL) and extracted with EtOAc (3 x 20 mL). The combined extracts were washed with water (2 x 50 mL), dried (brine, Na2S04), evaporated, and purified 15 by dry-flash column chromatography (silica gel, 10-90% EtOAc/hexane), to give 8a (1.00 g, 83%) as a cream powder: mp 120-121 °C; JH NMR (CDCI3) major rotamer 8 8.94 (br d, 2.9 Hz, 1 H), 8.52 (dd, J= 8.6,1.5 Hz, 1 H), 7.45-7.58 (m, 3 H), 7.25-7.40 (m, 3 H), 6.96 (br s, 1 H), 5.46 (s, 2 H), 4.40 (t, J= 4.7 Hz, 1 H), 3.84 (br ddd, J= 14.6,7.3,7.3 Hz, 1 H), 3.33 (ddd, J= 14.6, 8.2, 5.8 Hz, 1 H), 20 3.28,3.25 (2 x s, 3 H each), 1.65-1.95 (m, 2 H), 1.23 (br s, 9 H); 13CNMR (CDCI3) major rotamer 8 154.6, 153.6, 149.9,143.8,141.3,139.8,136.0,131.2, 128.7,128.0,127.0,123.4, 112.3,102.9, 93.3, 80.3, 70.9, 53.1, 52.7, 45.4, 31.2, 28.1; C26H31IN2O5 requires M^ 578.1278. Found 578.1257. 8-BenzyIoxy-6-j7V-(fert-butyIoxycarbonyl)-jV-(3-oxopropyI)amino]-5- iodoquinoline (9a). A solution of 8a (0.75 g, 1.30 mmol), Ts0H.H20 (0.12 g, 0.65 mmol) and water (3.75 mL) in acetone (38 mL) was stiixed at reflux for 2.25 h. Most of the acetone was evaporated and the residue was diluted with water (50 mL) and saturated, aqueous NaHCOs (5 mL) and extracted with EtOAc (3 x 20 30 mL). The combined extracts were washed with water (2 x 50 mL), dried (Na2S04), and evaporated to give 9a (0.68 g, 99%) as a pale yellow foam; !H 34 NMR (CDCI3) major rotamer 8 9.68 (s, 1 H), 8.97 (dd, J** 4.2,1.5 Hz, 1 H), 8.51 (dd, J= 8.6,1.5 Hz, 1 H), 7.53 (dd, J= 8.6,4.2 Hz, 1 H), 7.47-7.55 (m, 2 H), 7.25-7.40 (m, 3 H), 6.87 (br s, 1 H), 5.49 (s, 2 H), 4.17 (br dt, /= 14.5,7.1 Hz, 1 H), 3.59 (dt, J= 14.5,6.5 Hz, 1 H), 2.57 (br dd, J= 7.1, 6.5 Hz, 2 H), 1.23 (s, 9 5 H); 13C NMR (CDC13) major rotamer 8 200.3,154.8, 153.4,150.0, 143.0, 141.0, 139.7,135.9,131.0,128.6,127.9,127.0,123.4,112.1, 93.1, 80.7,70.7,42.9,42.5, 27.9; C24H25IN2O4 requires M1" 532.0859. Found 512.0862. 6-[iV-(3-Acetoxy-2-propenyI)-iV-(fer^butyloxycarbonyl)amino]-8-benzyloxyr5-10 iodoqninoline (10a). A mixture of ?a (0.62 g, 1.16 mmol), Et3N (0.40 mL, 2.87 mmol), AC2O (0.25 mL, 2.65 mmol), DMAP (14 mg, 0.11 mmol), andTHF (12 mL) was stirred at reflux for 2 h. Further EtaN (0.80 mL, 5.74 mmol), AC2O (0.50 mL, 5.3 mmol), and DMAP (10 mg, 0.08 mmol) were added and heating was continued for a further 2 h. The solvent was evaporated, and the residue was 15 diluted with pH 7.4 phosphate buffer (50 mL) and extracted with EtOAc (3 x 20 mL). The combined extracts were washed with water (50 mL), dilute aqueous NaHC03 (50 mL), and water (50 mL) before being dried (brine, NaiSO^, and evaporated. The residue was purified by dry-flash column chromatography (silica gel, 10-80% EtOAc-hexane) to give 10a (0.54 g, 81%) as a white foam, which 20 contained a 1:4 mixture of Z and E isomers: *H NMR (CDCI3) major rotamer 8 8.94 (br s, 1 H), 7.45-7.55 (m, 3 H), 7.27-7.40 (m, 3 H), 6.84-7.12 (m, 2 H), 5.36-5.58 (m, 2.8 H), 4.91 (ddd, /= 7.6, 6.5, 5.9 Hz, 0.2 H), 4.57 (dd, J= 15.0, 5.9 Hz, 0.2 H), 4.39 (dd, J= 14.7,6.8 Hz, 0.8 H), 4.06 (dd, J= 15.0, 7.6 Hz, 0.2 H), 3.86 (dd, J= 14.7,7.9 Hz, 0.8 H), 2.08 (s, 2.4 H), 1.88 (s, 0.6 H), 1.57 (br s, 25 1.8 H), 1.26 (br s, 7.2 H); 13C NMR (CDCI3) major rotamer 8 167.4,167.0,154.5, 149.8,154.3,149.8,153.3,153.1,142.8,140.9,139.7,138.8, 139.7,143.1,135.8, 130.9,136.0,127.8,126.8,126.7, 128.4,123.2,112.1,112.0,109.1,108.2, 93.5, 93.1, 80.9, 80.4, 70.8, 70.7,46.4,42.7,27.9, 28.1,20.3, 20.1; C26H27IN20s requires M1" 574.0965. Found 574.0962. ' WO 02/059122 PCT/NZ02/00005 l-(Acetoxymethyl)-5-benzyloxy-3~(tert-butyioxycarbonyl)-2,3-dihydro-lI?-pyrrolo [3,2-/] quinoline (11). A solution of 10a (0.54 g, 0.94 mmol), AIBN (15 mg, 0.09 mmol), and BU3S11H (0.32 g, 1.13 mmol) in benzene (45 mL) was stirred at reflux under nitrogen for 5.5 h. The solvent was evaporated, the residue was 5 triturated witib. pentane, and the precipitate was collected by filtration to give 11 (0.32 g, 77%), which crystallized from MeOH as fluorescent pale yellow rectangular plates: mp 172-173 °C; ^NMRCCDCfc) 8 8.82 (dd,J=4.1,1.4 Hz, 1 H), 8.14 (dd, J= 8.4,1.4 Hz, 1 H), 8.07 (br s, 1 H), 7.55 (br s, 2 H), 7.41 (dd, J = 8.4,4.1 Hz, 1 H), 7.36 (dd, J= 7.3,7.3 Hz, 2 H), 7.30 (tt, J= 7.3,2.4 Hz, 1 H), 10 5.44, 5.39 (2 x d, J= 12.5 Hz, 1 H each), 4.42-4.52 (m, 1 H), 4.05-4.14 (m, 2 H), 3.82-3.93 (m, 2 H), 2.08 (s, 3 H), 1.57 (s, 9 H); 13CNMR (CDC13) 8 171.0,155.2, 152.3, 146.9,142.0 (br), 137.0,136.3,131.1,128.5,127.9, 127.7,126.0,122.1, 113.3 (v. br), 100.4 (br), 81.4 (br), 70.7,65.8,52.6,37.7,28.4,20.9. Anal. Calcd for C26H28N205: C, 69.63; H, 6.29; N, 6.25. Found: C, 69.46; H, 6.27; N, 6.30.

-Benzyloxy-3-(ter/-butyloxycarbonyl)-l-(hydroxymethyl)-2,3-dihydro-lff-pyrrolo[3,2-f\quinoline (12). A mixture of 11 (0.22 g, 0.50 mmol), CS2CO3 (0.42 g, 1.29 mmol), and EtOH-water (2:1, 6 mL) was stirred at reflux for 30 min. The mixture was diluted with EtOAc (30 mL) and dilute aqueous NaHCOs (50 mL). 20 The separated aqueous phase was extracted with EtOAc (30 mL). The combined extracts were washed with water (3 x 50 mL), dried (brine, Na2S04), and evaporated to give 12 (0.19 g, 95%), which crystallized from MeOH as tiny white needles: mp 170-171 °C; *H NMR (CDC13) 8 8.54 (br s, 1 H), 7.99 (br d, /= 8.0 Hz, 1 H), 7.91 (br s, 1 H), 7.55 (d, J= 6.6 Hz, 2 H), 7.20-7.40 (m, 4 H), 5.29 (s, 2 25 H), 4.00-4.22 (m, 2 H), 3.65-3.78 (m, 3 H, H-l), 3.23 (br s, 1 H), 1.56 (s, 9 H); 13C NMR (CDCI3) 8 154.4,152.5 (br), 146.2 (br), 142.2 (v. br), 136.3,136.2, 131.3, 128.5,128.0 (v. br), 127.9,125.9,121.6,114.7 (v. br), 100.4 (br), 81.0 (br), 70.7, 64.6, 52.3,40.9 (br), 28.4. Anal. Calcd. for C24H26N2O4.H2O: C, 67.91; H, 6.65; N, 6.60. Found: C, 68..16; H, 6.47; N, 6.71. 36 S-Benzyloxy-l-tmethylsulfonyloxymethyty-S-^ert-butyloxycarbonyl^S-dihydro-LET-pyrroIo [3,2^ quinoline (13). MsCl (0.06 mL, 0.7 mmol) was added to a cooled (ice-water) solution of 12 (0.17 g, 0.41 mmol) and Et3N (0.2 mL, 1.4 mmol) in CH2CI2 (3 mL) and the mixture was stirred for 30 min. The CH2CI2 was 5 evaporated and the residue was stirred with water (25 mL) for 10 min. Hie mixture was extracted with EtOAc (2 x 25 mL). The combined extracts were washed with water (2 x 50 mL), dried (NaaSO^), and evaporated to give 13 (0.17 g, 86%), which crystallized from MeOH as tiny cream needles: mp 156-157 °C; *HNMR(CDCI3) 8 8.80 (dd, 4.2,1.4 Hz, 1 H), 8.02 (dd, J= 8.7,1.4Hz, 1 H), 10 7.97 (br s, 1 H), 7.55 (br d, J= 6.9 Hz, 2 H), 7.41 (dd, J= 8.7,4.2 Hz), 7.25-7.38 (m, 3 H), 5.40 (s, 2 H), 4.46 (dd, J= 9.8,3.7 Hz, 1H), 3.93-4.24 (m, 4 H), 2.90 (s, 3 H), 1.57 (s, 9 H); 13C NMR (CDCI3) 8 155.6, 152.1, 147.0,141.0 (v. br), . 137.1,136.1, 130.5,128.4,127.9, 127.6 (br), 125.7,122.3,112.7 (v. br), 100.3, 81.6 (br), 70.7,69.9,52.0,38.2 (br), 37.4,28.3. Anal. Calcd for C2sH28N206S: C, 15 61.97; H, 5.82; N, 5.78; S, 6.62. Found: C, 62.15; H, 5.96; N, 5.88; S, 6.54.

-Benzyloxy-3-(toT-butyloxycarbonyI)-l-(chloromethyl)-2,3-dihydro-li/-pvrrolo[3.2-/]quinoline (14). Method 1. A mixture of 13 (50 mg, 0.10 mmol), LiCl (25 mg, 0.59 mmol), and DMF (0.25 mL) was stirred at 80 °C for 1 h, before 20 ice (3 g) was added. The precipitate was removed by filtration, washed with water, and taken up in EtOAc (20 mL). This solution was washed with water (20 mL), dried (Na2S04), and evaporated to give 14 (39 mg, 89%), which crystallized from MeOH as fluorescent cream needles: mp 178-179 °C; LHNMR (CDCI3) 8 8.82 (dd, 4.2,1.5 Hz, 1 H), 8.05 (br s, 1 H), 7.99 (br d, J= 8.4 Hz, 1 H), 7.55 25 (br s, 2 H), 7.41 (dd, 8.4,4.2 Hz, 1 H), 7.35 (dd, J= 7.3,7.3 Hz, 2 H), 7.30 (tt, 7.3,2.4 Hz, 1 H), 5.42, 5.38 (2 x d, J= 12.4 Hz, 1 H each), 4.23 (br d, J= 11.7 Hz, 1 H), 4.12 (dd, J= 11.7, 8.9 Hz, 1 H), 3.92 (dddd, J= 10.1, 8.9,3.2,2.6 Hz, 1 H), 3.81 (dd, 11.1,3.2 Hz, 1 H),3.45 (dd, 11.1,10.1 Hz, 1 H), 1.56 (s, 9 H); 13CNMR(CDC13)8 155.5,152.3, 146.9, 141.9 (br), 137.1,136.3, 130.3, 128.5,127.9, 127.7 (br), 125.6,122.2,113.4 (v. br), 100.4 (br), 81.6 (br), 70.8, 37 53.0,46.3,41.1,28.4. Anal. Calcd. for C24H25CIN2O3: C, 67.84; H, 5.93; CI, 8.34; N, 6.59. Found: C, 67.85; H, 5.94; N, 6.68; CI, 8.26. 8-BenzyIoxy-6-[iV-(to'/'-butyIoxycarbonyl)-Ar-(3-chIoro-2-propenyl)aniino]-5-5 iodoquinoline (15a). NaH (60% dispersion in oil, 0.26 g, 6.5 mmol) under nitrogen was washed with pentane (3x2 mL), cooled (ice-water), and treated with a solution of 7a (2.80 g, 5.88 mmol) in DMF (2-8 mL) over 5 min. The cooling bath was removed and the mixture was allowed to stir for 30 min, by which time the solution was deep yellow and effervescence had ceased. 1,3-10 Dichloropropene (0.98 g, 8.82 mmol) was added and the mixture was stirred for 86 h. The mixture was diluted with water (150 mL) and extracted with EtOAc (4 x 25 mL). The combined extracts were washed with water (3 x 100 mL), dried (brine, NaaSO^, and evaporated. The residue was triturated with pentane and the precipitate was collected by filtration to give 15a (3.02 g, 93%) as a tan powder: 15 mp 115-135 °C containing a 1:1 mixture of Z and E isomers; *H NMR (CDCI3) major rotamer 8 8.95 (br s, 1 H), 8.50 (dd, J— 8.4,2.5 Hz, 1 H), 7.46-7.55 (m, 3 H), 7.27-7.41 (m, 3 H), 6.79-6.96 (m, 1 H), 5.30-6.03 (m, 4 H), 4.54 (dd, J= 15.5, 5.6 Hz, 0.5 H), 4.38 (dd, J= 14.8, 6.8 Hz, 0.5 H), 4.18 (dd, J= 15.5, 6.9 Hz, 0.5 H), 3.79 (dd, J= 14.8, 7.8 Hz, 0.5 H), 1.23-1.82 (m, 9 H); 13C NMR (CDCI3) 20 major rotamer 8 154.7, 155.2,153.6,153.3, 150.2, 150.1, 143.2,142.8,141.2, 140.2, 136.2,136.0, 131.13,131.08,128.79,128.73,128.12,127.99, 127.2,126.6, 126.98,126.90,123.5,123.4, 122.0,121.1, 112.2,111.9, 93.65, 93.58, 80.90, 80.85,71.0,70.9,48.8,45.4,28.4,28.1. C24H24CIIN2O3 requires M+" 550.0520, 552.0491. Found 550.0536,552.0503. Purification of the mother liquors by dry-25 flash column chromatography (silica gel, 10-60% EtOAo-hexane) gave further 15a (0.14 g, 4%).

Compound 14 by Method 2. A solution of 15a (3.00 g, 5.45 mmol), AIBN (89 mg, 0.54 mmol), and Bu3SnH (1.75 g, 6.0 mmol) in benzene (270 mL) was heated 30 at reflux under nitrogen for 3 h. The benzene was evaporated, the residue was WO 02/059122 PCT/NZ02/00005 38 triturated with pentane, and the precipitate was collected by filtration to give 14 (2.21 g, 95%), identical to the material prepared above.

Compound 14 by Method 3. A mixture of 12 (19 mg, 0.047 mmol), Ph3P (37 5 mg, 0.14 mmol) and CH2CI2 (0.4 mL) was treated with CCI4 (0.05 mL, 0.52 mmol), and the mixture was stirred under nitrogen for 4 h. The mixture was diluted with dilute aqueous NaHC03 (5 mL) and extracted with EtOAc (3x5 mL). The combined extracts were dried (Na2S04), evaporated, and purified by dry-flash column chromatography (silica gel, 10-90% EtOAc/hexane) to give 14 10 (20 mg, 100%) identical with the material prepared above. 3-(terf-Butyloxycarbonyl)-l-(chloromethyI)-5-hydroxy-2,3-dihydro-Lff- pyrrolo[3,2^quinoline (16). A cooled (ice-water) mixture of 14 (0.11 g, 0.27 mmol), 10% Pd/C (55 mg), and THF (5 mL) under nitrogen was treated with 25% 15 aqueous NH4HCO3 (0.67 mL). The mixture was stirred at 0 °C for 6 h, and was then diluted with EtOAc (20 mL), dried (Na2S04), filtered through Celite, evaporated, and purified by dry-flash column chromatography (silica gel, 10-50% EtOAc/hexane) to give 16 (39 mg, 44%) as a white solid: mp 148-149 °C; !H NMR (CDCI3) 5 8.61 (dd, J= 4.2,1.2 Hz, 1 H), 8.01 (dd, J= 8.5,1.2 Hz, 1 H), 20 7.83 (br s, 1 H), 7.41 (dd, 8.5,4.2 Hz, 1 H), 4.26 (dd, J= 11.8,2.2 Hz, 1 H), 4.14 (dd, J= 11.8, 8.5 Hz, 1 H), 3.93 (dddd, 9.8, 8.5, 3.2,2.2 Hz, 1 H), 3.80 (dd, 11.1,3.2 Hz, 1 H), 3.46 (dd, J= 11.1,9.8 Hz, 1 H), 1.61 (s, 9 H); 13C NMR (CDCI3) 5 153.5,152.3,• 145.3,142.4 (br), 135.0,130.6,124.9, 122.6,112.4 (v. br), 100.0, 81.7 (br), 53.0,46.5,40.9,28.4. C17H19CIN2O3 requires M** 25 334.1084, 336.1055. Found 334.1081, 336.1058.

-Benzyloxy-l-(chloromethyl)-3-(5,6,7-trimethoxyindoi-2-ylcarbonyI)-2,3-dihydro-l_fiT-pyrrolo[3,2-/]quinoline (17a). A suspension of 14 (0.65 g, 1.53 mmol) in dioxane (40 mL) was saturated with HC1, allowed to stand for 1 h, and 30 evaporated. 5,6;7-Trimethoxyindole-2-carboxylic acid (0.38 g, 1.53 mmol), EDCI (0.88 g, 4.6 mmol) and DMA (25 mL) were added to the remaining green-yellow 39 solid, and the red mixture was stirred at room temperature for 39 h. The mixture was poured into a mixture of ice (60 g) and pH 7.4 phosphate buffer (60 mL). The precipitate was removed by filtration, washed with water, and taken up in EtOAc (60 mL). This solution was washed with water (3 x 50 mL), dried (brine, 5 Na2S04), and evaporated. The remaining oil was triturated with Et20. The precipitate was collected by filtration, purified by flash column chromatography (silica gel, EtOAc), and triturated with Et20 to give 17a (0.3 8 g, 44%) as a pale yellow solid: mp 182-184 °C; ^NMR (CDC13) 6 9.59 (s, 1 H), 8.84 (dd, J= 4.2, 1.6 Hz, 1 H), 8.37 (s, 1 H), 7.95 (dd, J~ 8.5,1.6 Hz, 1 H), 7.58 (br d, J= 7.2 Hz, 10 2 H), 7.38 (dd, J= 8.5,4.2 Hz, 1 H), 7.36 (dd, J= 7.3,7.2 Hz, 2 H), 7.30 (t, J-13 Hz, 1 H), 6.93 (d, J= 2.2 Hz, 1 H), 6.84 (s, 1 H), 5.48, 5.42 (2 x d, J= 12.5 Hz, 1 H each), 4.69 (dd, J= 10.8,1.9 Hz, 1 H), 4.57 (dd, J= 10.8, 8.5 Hz, 1 H), 4.06, 3.93, 3.90 (3 x s, 3 H each), 4.02 (dddd, J= 10.3, 8.5, 3.2,1.9 Hz, 1 H), 3.83 (dd, J- 11.4,3.2 Hz, 1 H), 3.42 (dd, J= 11.4,10.3 Hz, 1 H); I3C NMR (CDC13) 8 15 160.5,155.3,147.8,150.2,142.3,140.6,138.8,138.2,129.5,125.1,123.5,136.4, 130.4,128.6, 128.0,127.7,125.6,122.3,115.3,1.06.7,102.3,97.6, 70.8, 61.4, 61.1, 56.2, 55.1, 45.9, 42.5. C31H28aN305 requires M+H 558.1796, 560.1766. Found (FAB) 558.1770, 560.1786. Anal. Calcd for C3iH28ClN3Os: C, 66.72; H, 5.06; N, 7.53. Found: C, 66.96; H, 5.36; N, 7.50. l-(Chloromethyl)-5-hydroxy-3-[(5,6,7-trimethoxyindol-2-yl)carbonyl]-2,3-dihydro-Iff-pyrrolo [3,2-/| quinoline (18a). Method 1. THF (10 mL) then 25% aqueous NH4HCO3 (1.1 mL) were added to a cooled (ice-water) mixture of 17a (0.25 g, 0.45 mmol) and 10% Pd/C (0.13 g) under nitrogen. The mixture was 25 stirred at 0 °C for 7.5 h, and was then filtered through Celite. The Celite was washed with a solution of concentrated HC1 (2 mL) and MeOH (40 mL) and then with CH2Cl2-MeOH (3:1,40 mL). The combined filtrates were diluted with water (40 mL) and CH2CI2 (30 mL) and neutralized with pH 7.4 phosphate buffer. The lower layer was separated then diluted with MeOH (20 mL) and warmed to 30 dissolve the suspended solid. The aqueous phase was extracted with CH2CI2 (2 x 20 mL). The extracts were combined, washed with water (100 mL), dried 40 (NaaSOO, and concentrated to a volume of 20 mL. The concentrate was diluted with MeOH (20 mL) and was concentrated to a volume of 10 mL. The precipitate was removed by filtration and washed with MeOH to give 18a (0.14 g, 66%) as a pale yellow microcrystalline solid: mp > 230 °C; ]H NMR [(CD3)2SO] 8 11.50 (d, 5 /= 2.1 Hz, 1H), 10.03 (br s, 1 H), 8.76 (dd, J= 4.1,1.3 Hz, 1 H), 8.40 (dd, J= 8.4,1.3 Hz, 1 H), 7.97 (s, 1 H), 7.56 (dd, J= 8.4,4.1 Hz, 1 H), 7.09 (d, 2.1 Hz, 1 H), 6.97 (s, 1 H), 4.77 (dd, /= 11.0,9.3 Hz, 1 H), "4.48 (dd, J= 11.0,2.0 Hz, 1 H), 4.25 (dddd, /= 9.3,3.9,3.3,2.0 Hz, 1 H), 4.03 (dd, J= 10.6,3.3 Hz, 1 H), 3.93,3.82,3.80 (3 * s, 3 H each), 3.89 (dd, J= 10.6,3.9 Hz, 1 H); 13CNMR .

((CD3)2SO) 8 160.3,153.9,146.3, 149.1,142.7,139.9,139.0,136.0,130.7, 125.4, 124.8,123.1,131.6,122.4, 114.6,106.2, 102.8,98.0,61.0,60.9, 55.9, 55.0,47.6, 40.5. Anal. Calcd for C24H22CIN3O5: C, 61.61; H, 4.74; CI, 7.58; N, 8.98. Found: C, 61.50; H, 4.98; N, 8.84.

Compound 18a by Method 2. A solution of 16 (0.14 g, 0.43 mmol) in dioxane (9 mL) was saturated with HC1, allowed to stand for 1 h, and evaporated. 5,6,7-Trimethoxyindole-2-carboxylic acid (0.11 g, 0.43 mmol), EDCI (0.25 g, 1.28 mmol) and DMA (5 mL) were added to the remaining yellow solid, and the red mixture was stirred at room temperature for 22 h. The mixture was poured into a mixture of ice (20 g) and pH 7.4 phosphate buffer (20 mL). The precipitate was removed by filtration, washed with water, and taken up in CH2Cl2-MeOH (2:1,30 mL). The CH2CI2 was boiled off, the remaining mixture was cooled in ice, and the precipitate was removed by filtration to give 18a (18 mg, 9%) identical to the material prepared above.

Similarly were prepared: l-(Chloromethyl)-3-({5-[2-(dimethyIamino)ethoxy]-5-hydroxyindol-2-yl} carbonyI)-2,3-dihydro-llJ-pyrrolo \3,2-j\ quinoline (18b).

A suspension of 14 (0.20 g, 0.47 mmol) in cooled (0 °C) dioxane (5 mL) was saturated with HC1, allowed to warm to r.t. over 2 h and evaporated. 5-[2- 41 (DiirLethylamino)ethoxy]-l-JHr-indole-2-carboxylic acid hydrochloride (0.13 g, 0.47 mmol) [Milbank et al., J. Med. Chem., 1999,42, 649], EDC1 (0.27 g, 1.42 mmol) and DMA (3 mL) ware added to the remaining yellow solid, and the red mixture was stirred at r.t. for 20 h. The mixture was partitioned between EtOAc 5 and 5% NaHC03 solution. The aqueous layer was extracted with EtOAc (x3). The EtOAc extracts were dried (brine, Na2S04). Flash chromatography (Alumina, EtOAc/MeOH; 49:1, then 9:1) gave 2-[(2-{[5-(benzyIoxy)-l-(chloromethyl)-l,2-dihydro-3//-pyrrolo [3,2-f\quinolin-3 -yl] carbonyl} -1 i7-indol-5-yl)oxy] -Nfl-dimethylethanamine (17b) (0.22 g, 84%) as a yellow solid: mp 176-179 °C; [H _ 10 NMR [(CD3)2SO] 8 11.68 (s, 1 H), 8.79 (dd, 4.1,1.5 Hz, 1 H), 8.41 (dd, J= 8.6,1.5 Hz, 1 H), .8.29 (s, 1 H), 7.56 (m, 3 H), 7.40 (m, 4 H), 7.17 (d, J= 2.3 Hz, ' 1 H), 7.11 (d, /= 1.5 Hz, 1 H), 6.92 (dd, J= 9.0,2.4 Hz, 1 H), 5.32 (s, 2 H), 4.82 (dd, J= 10.7,9.6 Hz, 1 H), 4.58 (dd, 10.9,2.1 Hz, 1 H), 4.32 (m, 1 H), 4.05 (t, J= 5.7 Hz, 2 H), 4.04 (m, 1 H), 3.93 (dd, J= 11.2, 6.9 Hz, 1 H), 2.65 (t, J= 5.8 15 Hz, 2 H), 2.23 (s, 6H); 13C NMR [(CD3)2SO] 8 160.3, 154.5, 153.0, 147.3, 142.3, 137.4,136.7,131.6,131.3,130.6,128.4, 127.9,127.7,127.4, 125.1, 122.4,116.2, 116.0, 113.1,105.5,103.1,102.0, 70.0, 66.9, 66.2, 57.8, 54.9,47.7,45.5,40.7.

THF (8 mL) then HC02NH{ (0.23 g, 3.6 mmol) in H20 (1 mL) were added to 20 cooled (0 °C) mixture of 17b (0.20 g, 0.36 mmol) and 10% Pd/C (0.1 g) under N2. The mixture was stirred at 0 °C for 14 h, and was then filtered through Celite. The Celite was washed with CH2Cl2/H20. The aqueous layer was extracted with CH2C12 (x3). The CH2C12 extracts were dried (brine, Na2S04) and passed through a short plug of silica gel to give 18b (0.16 g, 93%) as a yellow solid: mp 209-215 25 °C; *H NMR [(CD3)2SO] 8 11.66 (s, 1 H), 10.02 (bs, 1 H), 8.76 (dd, .7=4.1,1.4, 1 H), 8.41 (dd, J= 8.5,1.3,1 H), 8.07 (s, 1 H), 7.56 (dd, J= 8.5,4.1,1 H), 7.40 (d, J= 8.9,1 H), 7.17 (d, J=2.2,1 H), 7.11 (d, J= 1.2,1 H), 6.93 (dd, J=8.9, 2.3,1 H), 4.82 (dd, 10.7,9.6,1 H), 4.57 (dd, /= 11.0,2.1,1 H), 4.29 (m, 1 H), 4.06 (t, J= 5.9,2 H), 4.04 (m, 1H), 3.91 (dd, /= 11.1,7.2,1 H), 2.64 (t, J= 5.8, 30 2 H), 2.28 (s, 6 H); 13C NMR [(CDs^SO] 8 160.3, 153.9,153.0,146.4,142.8, 42 136.1,131.6,130.7,127.4, 124.8, 124.7,122.5, 116.0,114.6,113.1, 105.5,103.1, 103.0,66.1, 57.8, 54.9,47.7,45.5,40.7. l-(Chloromethyl)-3-((2£)-3-{4-[2-(dimethylainino)etlioxy]phenyl}-2-5 propenoyl)-5-hydroxy-2,3-dihydro-lJ/-pyrroIo[3,2:/]qumoline (18c).

A suspension of 14 (0.20 g, 0.47 mmol) in cooled (0 °C) dioxane (5 mL) was saturated with HC1, allowed to warm to r.t. over 1 h and evaporated. (E)-4-[2-(Dimethylamino)ethoxy]ciimamic acid hydrochloride (0.13 g, 0.47 mmol) [Atwell et al., J. Med. Chem., 1999,42,3400], EDCI (0.27 g, 1.42 mmol) and DMA (3 . 10 mL) were added to the remaining yellow solid, and the red mixture was stirred at r.t. for 30 h. The mixture was partitioned between CH2CI2 and 5% NaHCC>3 solution. The aqueous layer was extracted with CH2CI2 (x3). The CH2CI2 extracts were dried (brine, Na2SC>4). Flash chromatography (Alumina, EtOAc/MeOH; 49:1, then 24:1) gave 2-(4-{(l£)-3-[5-(benzyloxy)-1 -(chloromethyl)-1,2-dihy dro-15 3 i?-pyrrolo [3,2-/] quinolin-3 -yl]-3 -oxo-1 -propenyl} phenoxy)-JV,./V- dimethylethanamine (17c) (0.18 g, 70%) as a yellow solid: mp 172-175 °C; lH NMR [(CDskSO] 8 8.76 (dd, J= 4.1,1.4,1 H), 8.47 (bs, 1 H), 8.35 (dd, J= 8.5, 1.4,1 H), 7.76 (d,J= 8.7,2 H), 7.67 (d, J= 15.3,1 H), 7.58 (d, J= 7.3, 2 H), 7.54 (dd, J= 8.5,4.1,1 H), 7.44 (t, J= 7.2,2 H), 7.37 (t, J= 7.2,1 H), 7.08 (d, J= 20 15.3,1 H), 7.02 (d, J= 8.7,2 H), 5.31 (s, 2 H), 4.55 (dd, J= 10.7,9.5,1 H), 4.44 (dd, J= 10.9,2.5,1 H), 4.30 (m, 1 H), 4.11 (t, J= 5.8,2 H), 3.99 (dd, 11.0, 3.0,1 H), 3.91 (dd, J= 11.2, 7.2,1 H), 2.64 (t, J= 5.7,2 H), 2.23 (s, 6 H); 13C NMR [(CD3)2SO] 8 164.1,160.1,154.6,147.1,142.6,142.2,137.2,136.7,131.1, 130.1,128.3,127.83, 127 78,127.3, 125.1, 122.3,116.9, 115.7,114.7, 101.6, 25 70.0,65.9,57.5, 52.9,47.8,45.4,40.1.

A solution of 17c (0.56 g, 1.03 mmol) was dissolved in CF3COOH (15 mL) and refluxed for 48 h. CF3COOH was evaporated and the residue was partitioned between CH2CI2 and cold 5% NaHC03 solution. The aqueous layer was extracted 30 with CH2CI2 (x3). The CH2CI2 extracts were dried (brine, Na2SC>4). Flash chromatography (CH2Cl2/MeOH/NH3; 95:5:trace) gave 18c (0.16 g, 34%) as a 43 yellow solid: mp 174-180 °C; lE NMR [(CD3)2S03 5 9.96 (bs, 1 H), 8.73 (dd, J= 4.0,1.3,1 H), 8.36 (dd, /= 8.4,1.3,1 H), 8.18 (bs, 1 H), 7.77 (d, /= 8.7,2 H), 7.66 (d,/= 15.2,1 H), 7.54 (dd, /= 8.5,4.1,1 H), 7.08 (d, J= 15.4,1H), 7.02 (d, J= 8.7, 2 H), 4.54 (dd, J= 10.7,9.5,1 H), 4.44 (dd, J= 11.0,2.5,1 H), 4.28 (m, 1 5 H), 4.11 (t, /= 5.7,2 H), 4.00 (dd,/= 11.1,3.1,1 H), 3.88 (dd,/= 11.0,7.4,1 H), 2.64 (t, 5.8,2 H), 2.22 (s, 6 H). l-(Chloromethyl)-3-[(5-methoxyindol-2-yl)carbonyl]-5-hydro3Ey-2,3-dihydro-lfl-py rrolo [3,2-^] quinoline (18d).

A suspension of 14 (0.10 g, 0.24 mmol) in dioxane (15 mL) was saturated with HC1, stirred at r.t. for 5 h and evaporated. 5-Methoxy-l -if-indole-2-carboxylic acid (0.054 g, 0.28 mmol), EDCI (0.23 g, 1.17 mmol) and DMA (5 mL) were added to the remaining yellow solid, and the red mixture was stirred at r.t. for 52 h. Hie mixture was partitioned between CH2CI2 and cold 5% KHCO3 solution. 15 The aqueous layer was extracted with CH2CI2 (x3). The CH2CI2 extracts were dried (brine, Na2SC>4). Flash chromatography (EtOAc/petroleum ether; 7:3) gave 5 -(benzyloxy)-1 -(chloromethyl)-3-[(5 -methoxy-l/f-indol-2-yl)carbonyl]-2,3 -dihydro-1 f/-pyrrolo[3,2-/Jquinoline (17d) (0.11 g, 98%) as a yellow solid : mp 186-189 °C; ^NMR (CDC13) 8 9.55 (s, 1 H), 8.88 (dd, J= 4.2,1.7,1H), 8.37 (s, 20 1 H), 7.99 (dd, J= 8.3,1.6,1 H), 7.56 (d, 7.3,2 H), 7.42 (dd, /= 8.3,4.1,1 H), 7.33 (m, 4 H), 7.10 (d, 2.3,1 H), 6.99 (m, 2 H), 5.48 (d, J= 12.5,1 H), 5.42 (d, J= 12.6,1 H), 4.74 (dd, /= 10.9,2.0,1 H), 4.61 (dd, J~ 10.6, 8.7,1 H), 4.05 (m, 1 H), 3.85 (s, 3 H), 3:84 (dd, 11.2,4.1,1 H), 3.45 (dd, 11.0,10.5, 1 H); 13CNMR(CDC13) 8 160.7,155.4, 154.7,147.9, 142.4, 138.4, 136.4,131.4, 25 130.5, 130.2,128.6,128.2,128.0, 127.7, 125.2,122.4,117.0,115.4, 112.7,106.2, 102.5, 102.4, 70.9, 55.7, 55.2,45.9,42.6.

THF (6 mL) then HCO2NH4 (0.14 g, 2.21 mmol) in H2O (0.7 mL) were added to cooled (0 °C) mixture of 17d (0.11 g, 0.22 mmol) and 10% Pd/C (0.05 g) under 30 N2. The mixture was stirred at 0 "C for 5 h, and was then filtered through Celite. The Celite was washed with CH2CI2/H2O. The aqueous layer was extracted with 44 CH2CI2 (x3). The CH2C12 extracts were dried (brine, Na2SC>4) and CH2CI2 evaporated. Precipitation firom CHaCfe/MeOH gave 18d (0.077 g, 89%) as a grey solid: mp 224-227 °C; JH NMR [(CD3)2SO] 5 11.66 (s, 1 H),10.02 (bs, 1 H), 8.77 (dd, 4.1,1.3,1 H), 8.41 (dd, J= 8.4,1.4,1 H), 8.07 (s, 1 H), 7.57 (dd, J= 8.4, 5 4.1,1 H), 7.40 (d, J= 9.0,1 H), 7.16 (d, /== 2.4,1 H), 7.12 (d, J= 1.6,1 H), 6.92 (dd, J= 8.9,2.3,1 H), 4.82 (dd, J= 10.8,9.4,1 H), 4.57 (dd, J= 11.0,2.3,1 H), 4.30 (m, 1 H), 4.04 (dd, J= 11.1,3.3,1 H), 3.91 (dd;/= 11.1,7.2, 1 H), 3.78 (s, 3 H). l-(Chioromethyl)-3-[(2£)-3-(4-niethoxyphenyI)-2-propenoyl]-5-hydroxy-2,3-dihydro-lJT-pyrroIo [3,2-J\ quinoline (18e). < A suspension of 16 (0.10 g, 0.30 mmol) in dioxane (5 mL) was saturated with HC1, stirred at r.t over 5 h and evaporated. 4-Methoxycinuamic acid (predominantly trans) (0.064 g, 0.36 mmol), EDCI (0.29 g, 1.50 mmol) and DMA 15 (3 mL) were added to the remaining yellow solid, and the red mixture was stirred at r.t. for 3 h. The mixture was partitioned between CH2CI2 and cold 5% KHCO3 solution. The aqueous layer was extracted with CH2CI2 (x3). The CH2CI2 extracts were dried (brine, Na2S04). Flash chromatography (CEfeGh/MeOH; 93:7) followed by recrystallisation (CH2Cl2/Et20) gave 18e (0.02 g, 17%) as a yellow 20 solid: mp 208-211 °C; >H NMR [(CP&SO] 5 9.96 (bs, 1 H), 8.73 (d, 3.3,1 H), 8.35 (d, 7.7,1 H), 8.18 (bs, 1 H), 7.78 (d, J= 8.7,2 H), 7.67 (d, 15.3,1 H), 7.54 (dd, /= 8.5,4.1,1 H), 7.08 (d, 15.4,1 H), 7.01 (d, /= 8.7,2 H), 4.54 (dd, J= 10.3,9.5,1 H), 4.45 (m, 1 H), 4.27 (m, 1 H), 3.99 (dd, J= 11.1,3.2,1 H), 3.88 (dd, J= 11.1, 7.3,1 H), 3.82 (s, 3 H). C22H20CIN2O3 requires M+H 25 395.1163,397.1133. Found (FAB) 395.1161,397.1169. l-(Chloromethyl)-3-[(2£)-3-(3-hydroxy-4-methoxyphenyl)-2-propenoyi]-5-hydroxy-2,3-dihydro-3JHr-pyrrolo[3^l'^]qiiinoline(18f).

A suspension of 16 (0.10 g, 0.30 mmol) in dioxane (5 mL) was saturated with 30 HC1, stirred at r.t. over 5 h and evaporated. 3-Hydroxy-4-methoxycinnarnic acid (predominantly trans) (0.070 g, 0.36 mmol), EDCI (0.29 g, 1.50 mmol) and DMA WO 02/059122 PCT/NZ02/00005 45 (3 mL) were added to the remaining yellow solid, and the red mixture was stirred at r.t. for 3 h. The mixture was partitioned between CH2CI2 and cold 5% KHCO3 solution. The aqueous layer was extracted with CH2CI2 (x3). The CH2CI2 extracts were dried (brine, Na2S04). Flash chromatography (CH2Cl2/MeOH; 93:7) 5 followed by recrystallisation (CH2Cl2/Et20) gave 18f (0.01 g, 8%) as a yellow solid: mp 215-218 °C; 'HNMR [(CD3)2SO] 5 9.96 (bs, 1 H), 9.13 (s, 1 H), 8.73 (dd, J= 4.1,1.4,1 H), 8.36 (dd, J= 8.5,1.4,1 H), 8.'17 (bs, 1 H), 7.57 (d, J= 15.3,1 H), 7.54 (dd, J= 8.5,4.1,1 H), 7.25 (d, /.= 2.0,1 H), 7.20 (dd, 8.4, 2.0,1 H), 6.99 (d, 8.1,1 H), 6.96 (d, J= 15.0,1 H), 4.54 (dd, /= 10.5, 9.4,1 10 H), 4.44 (dd, J= 11.1,2.6,1 H), 4.00 (dd, J= 11.2,3.3,1 H), 3.88 (dd, /= 11.1, 7.5,1 H), 3.83 (s, 3H). C22H]935C1N204 requires M+H 411.1112. Found (FAB) 411.1127.

Example B. Preparation of 5-amino-1 -fchloromethvlV3-[Y5.6.7-trimethoxvindol-15 2-vDcarbonvl1-2.3-dihvdro-ljfj"-pvrrolor3.2-flauinoline 27 by the method of Scheme 4. 8-NitroquinoIine-6-carboxylic acid (19). This was prepared by the reported method [Jung et al., Eur. Pat. Appln. EP 581500 (1994); Chem Abstr, 1994,122, 205125] in 41% yield: mp (EtOAc) 258-263 °C; NMR [(CD3)2SO] 5 13.80 (v 20 br, 1 H), 9.16 (dd, J= 4.3,1.7 Hz, 1 H), 8.96 (d, /= 1.7 Hz, 1 H), 8.80 (dd, J= 8.4,1.6 Hz, 1 Hi, 8.63 (d, J = 1.7 Hz, 1 H), 7.84 (dd, J= 8.4,4.2 Hz, 1 H). 6-(te/**.-Butyloxycarbonylamino)-8-nitroqumolme (20) A mixture of 19 (4.82 g, 22.1 mmol), DPPA (6.99 g, 25.4 mmol) and Et3N (3.69 mL, 26.5 mmol) in 25 anhydrous t-BuOH (60 mL) was heated at reflux under N2 for 8 h. The mixture was concentrated under reduced pressure, and the residue was partitioned between CH2CI2 and 10% aqueous KHCO3. The organic phase was washed with 10% aqueous KHCO3, dried (Na2S04) and concentrated under reduced pressure, then chromatographed on silica gel. Elution with CH2Cl2/EtOAc (17:3), followed by 30 sequential crystallisation from MeOH/HaO andCHjCVpetroleum ether gave 20 (3.82 g, 60%): mp 134-135 °C; JH NMR [(CD3)2SO] S 10.09 (s, 1 H), 8.87 (dd, J 46 = 4.1,1.5 Hz, 1 H), 8.47 (dd, 8.5,1.6 Hz, 1 H), 8.33 (d, J= 1.9 Hz, 1 H), 8.25 (d, J= 2,2 Hz, 1 H), 7.65 ,m(dd, J = 2.2 Hz, 1 H), 7.65 (dd, 8.5,4.2 Hz, 1 H), 1.53 (s, 9 H). Anal. Calcd. for Ci4Hi5N304: C, 58.12; H, 5.23; N, 14.53. Found: C, 58.39; H, 5.21 ;N, 14.65%. 8-Ammo-6-(fert.-butyIoxycarbonylamino)quinoline (21)A solution of 20 (3.3 0 g, 11.4 mmol) in MeOH (50 mL) was hydrogenated over 10% Pd/C at 50 psi for 3 h. The resulting crude product was filtered through a column of silica gel in EtOPAc to give 21 (2.71 g, 92%): mp (i-Pr20/petroleum ether) 131-132 °C; !H NMR [(CD3)2SO] 8 9.39 (s, 1 H), 8.54 (dd, J~ 4.1,1.6 Hz, 1 H), 8.01 (dd, J= 8.3,1.5 Hz, 1 H), 7.36 (dd, J= 8.3,4.1 Hz, 1 H), 7.24 (d, J= 2.0 Hz, 1 H), 6.97 (d, J= 2.1 Hz, 1 H), 5.90,5.88 (2xs, 2 H, NH2), 1.50 (s, 9 H). Anal. Calcd. for Ci4Hi7N302: C, 64.85; H, 6.61; N, 16.20. Found: C, 64.60; H, 6.77; N, 16.19%. 6-(tert.-Butyloxycarbonyl)-8-(1.3-dioxo-l,3-dihydro-2H-isoindol-2-15 yI)quinoline (22). A mixture of 21 (1.53 g, 5.90 mmol), phthalic anhydride (1.05 g, 7.09 mmol) and DMAP (36 mg, 5 mol%), in anhydrous pyridine (15 mL) was heated with stirring at 80 °C for 1 h. The mixture was concentrated under reduced pressure, then AcOH (10 mL) and Ac20 (5 ml) were added and the mixture was stirred at 80 °C for a further 45 min. Concentration under reduced pressure, 20 followed by addition of aqueous KHC03, gave a solid that was chromatographed on silica gel. Elution with CH2Cl2/EtOAc (4:1) gave a crude product that was crystallized from CH2Cl2/iPr20 to give 22 (2.09 g): mp 217-218 °C (dec.); *H NMR [(CD3)2SO] 8 10.00 (s, 1- H), 8.67 (d, J= 3.2 Hz, 1 H), 8.39 (d, J= 8.1 Hz, 1 H, H-4), 8.24 (s, 1 H), 8.80-7.89 (m, 5 H), 7.53 (dd, J= 8.3,4.1 Hz, 1 H), 1.53 (s, 25 9 H). Anal. Calcd. for CjaHieNsO^ C, 67.85; H, 4.92; N, 10.79. Found: C, 67.87; H, 4.94; N, 10.87%.

-Bromo-6-(fert.-butyIoxycarbonylamino)-8-(l,3-dioxo-l,3~dihydro-lH-isoindol-2-yI)quinoKne (23). A mixture of22 (1.79 g, 4.6 mmol) andNBS (0.98 30 g, 5.5 mmol) in anhydrous CH3CN (50 mL) was stirred at reflux for 45 min., then concentrated under reduced pressure. The residue was dissolved in CH2C12, and 47 the solution was washed with aqueous Na2S205/NaHC03 and water (2x), dried (Na2S04) and concentrated to dryness. The residue was chromatographed on silica gel, and elution with CHaCk/EtO Ac (3:2) gave a crude product that was crystallized from EtOAc/iP^O to 23 (1.91 g, 89%): mp 210-211 °C (dec.); XH 5 NMR [(CD3)2SO] 5 9.19 (s, 1 H), 8.85 (dd, J-4.1,1.4 Hz, 1H), 8.65 (dd, J= 8.6,1.4 Hz, 1 H), 8.21 (s, 1 H), 8.07-7.92 (m, 4 H), 7.75 (dd, J= 9.7,4.2 Hz, 1 H), 1.50 (s, 9 H). Anal. Calcd. for C22Hi8BrN304: C, 56".42; H, 3.87; N, 8.98; Br, 17.06. Found: C, 56.49; H, 4.04; N, 8.86; Br, 16.87%. 5-Bromo-6-[N-(fejtf.-butyIoxycarbonyI)-jV-(3-chIoro-2-propen-l-yl)amino]-8-(l,3-dioxo-l,3-dihydro-lH-isoindol-2-yl)quinoIine (24). A solution of 23 (1.82 g, 3.89 mmol) in anhydrous DMF (20 mL) was treated at 0 °C under N2 with NaH (0.20 g, 5.00 mmol, 60% in oil), and then stirred at 25 °C for 45 min. The mixture was then cooled to 0 °C and 1,3-dichloropropene (1.11 mL, 11.7 mmol) was 15 added. The reaction mixture was' warmed to 25 °C, stirred for 4 h, and then diluted with CH2CI2 (200 mL). The solution was washed with 10% aqueous KHC03 and water (2x), then dried (Na2S04) and concentrated under high vacuum at 25 °C. The residue was chromatographed on silica gel, eluting with CH2CI2 then CTbCyEtQAc (17:3) to give 24 (1.62 g, 77%) as a foam that was used directly; 20 NMR [(CD3)2S03 (mixture of rotamers of E and Z alkenes) 8 8.94 (d, J= 4.0 0 Hz, 1H), 8.73 (d,/= 8.6 Hz, 1H), 8.12-7.93 (m, 5 H), 7.80 (dd,J= 8.6,4.2 Hz, 1 H), 6.50-6.35 (m, 1 H), 6.21-6.02 (m, 1 H), 4.62-4.06 (m, 2 H), 1,51,1.32 (2xs, 9 H). Anal. Calcd. for CzsH^BrClN^^O: C, 51.87; H, 4.35; N, 7.26. Found: C, 51.69; H, 3.87; N, 6.86%. 3-(ifert.-ButyIoxycarbonyl)-l-(cMoromethyl)-5-(l^-dioxo-l,3-dihydro-ijff-isoindol-2-yl)-2,3-dihydro-I/f-pyrrolo[3,2-fiquinoline (25). A mixture of 24 (1.96 g, 3.61 mmol) and catalytic AJBN (60 mg, 10 mol%) in anhydrous benzene (20 mL) was treated with Bu3SnH (1.16 mL, 4.3 3 mmol) and heated at reflux 30 under N2 for 3 h. The reaction mixture was concentrated under reduced pressure and the residue was chromatographed on silica gel. Elution with QHaCyEtOAc 48 (17:3), followed by crystallisation from Cl^Ck/petroleum ether gave 25 (1.28 g, 76%): mp 163-165 °C; ^NMR [(CD3)2SO] 8 8.70 (dd, J= 4.1,1.3 Hz, 1 H), 8.51 (dd, J= 8.6,1.4 Hz, 1 H), 8.45 (v br, 1 H), 8.06-7.90 (m, 4 H), 7.57 (dd, /= 8.5,4.1 Hz, 1 H), 4.44-4.34 (m, 1 H), 4.29 (t, J= 10.5 Hz, 1 H), 4.19-3.99 (m, 3 5 H), 1.54 (s, 9 H). Anal. Calcd. for C25H22CIN3O4: C, 64.72; H, 4.78; N, 9.06. Found: C, 64.76; H, 4.92; N, 9.03%. l-(Chloromethyl)-5-(l,3-dioxo-lj3-dihydro-2Jff-isoindol-2-yI)-3-[(5,6,7-trimethoxymdoI-2-yl)carbonyl]-2,3-dihydro~lH-pyrrolo[3,2-f|quinoline (26).

A solution of 25 (500 mg, 108 mmol) in dioxane at 10 °C was saturated with HC1 gas, allowed to stand at 20 °C for 1 h, then evaporated to dryness under reduced pressure below 30 °C. 5,6,7-Trimethoxymdole-2-carboxylic acid (298 mg, 1.19 mmol), EDCI (518 mg, 2.70 mmol) and anhydrous DMA (10 mL) were then added, and the mixture was stirred at 20 °C for 3 h. Addition of 10% aqueous 15 KHCO3 precipitated a solid that was chromatographed on silica gel. Elution with QfeCyEtOAc (1:1), followed by crystallisation firom Et0Ac/iPr20, gave 1-(chloromethyl)-5-(phthalimido)-3-[(5,6,7-trimethoxyindol-2-yI)carbonyl]-2,3-dihydro-iiJ-pyrrolo[3,2-f]quinoline 26 (392 mg, 61%): mp 189-191 °C; *H NMR [(CD3)2S0J 8 11.54 (s, 1 H), 8.77 (dd,J= 4.1,1.4 Hz, 1 H), 8.75 (s, 1 H), 8.59 20 (dd, J= 8.5,1.4 Hz, 1 H), 8.08-7.92 (m, 4 H), 7.61 (dd, J= 8.5,4.2 Hz, 1 H), 7.14 (d, J= 1.7 Hz, 1 H), 6.98 (s, 1 H), 4.89 (dd, J= 10.8,9.7 Hz, 1 H), 4.61 (dd, J= 11.0,2.3 Hz, 1 H), 4.55-4.44 (m, 1 H), 4.20-4.05 (m, 2 H), 3.94,3.83,3.81 (3xs, 3x3H). Anal. Calcd. for C32H25C1N406: C, 64.37; H, 4.22; N, 9.39. Found: C, 64.04; H, 4.28; N, 9.29%.

-Amino-l-(chlorometliyI)-3-[(5,6,7-trimethoxymdoI-2-yl)carbonyI3-2,3-dihydro-IiJ-pyrroIo[3,2-f|quinoIine (27). A solution of26 (160 mg, 0.27 mmol) in CH2CI2 (6 mL) was was diluted with EtOAc (8 mL) and treated immediately with hydrazine monohydrate (155 pL, 3.19 mmol). The reaction mixture was 30 stirred at 25 °C for 2 h, then diluted with CH2C12 (40 mL), washed with 10% aqueous Na2C03 (2x) and saturated aqueous NaCl (2x), dried (Na2S04) and 49 concentrated under reduced pressure below 30 °C. Chromatography on silica gel, eluting with CEfeCIa/EtOAc (1:1) gave 27 (81 mg, 65%): mp 225-227 °C; !H NMR [(CD3)2S03 S 11.44 (s, 1 H), 8.63 (dd, J= 4.1,1.4 Hz, 1 H), 8.25 (dd, J= 8.5,1.4 Hz, 1 H), 7.82 (s, 1 H), 7.47 (dd, /= 8.5,4.1 Hz, 1 H), 7.05 (d, J= 1.2 Hz, 5 1 H), 6.99 (s, 1 H), 6.18, 6.16 (2xs, 2 H), 4.70 (dd, J= 10.8,9.1 Hz, 1 H), 4.43 (dd, J= 11.0,1.8 Hz, 1 H), 4.16-4.08 (m, 1 H), 3.97 (dd, J= 11.0,3.3 Hz, 1 H), 3.94,3.82, 3.80 (3xs, 3x3 H), 3.76 (dd, J= 10.0,7.8'Hz, 1 H). Anal. Calcd. for C24H23CIN4O4: C, 61.74; H, 4.96; N, 12.00. Found: C, 61.51; H, 5.04; N, 11.69%.

Example C: Preparation of 5-f2-ammoethvlaimnoV3-R"5.6.7-trimethoxvindol-2-vDcarbonvll-1 -("chloromethvD-1.2-dihvdro-3g-benzrelindole dihvdrochloride 29 bv the method of Scheme 5. A mixture of 5-amino-l-(chloromethyl)-3-[5,6,7-trimethoxyindol-2-yl)carbonyl]-1,2-dihydro-3i7-benz[e]indole 28 [Atwell et al., J. Org. Chem. 1998, 63, 9414] (252 mg, 0.54 mmol), N-(tert-15 butyloxycarbonyl)aminoacetaldehyde (430 mg, 2.70 mmol) TsOH (10 mg) and microwave-dried powdered A4 molecular sieves (3 g) in DMA (3 mL) and MeOH (0.5 mL) was stirred at 20 °C under N2 with the exclusion of light for 48 h. NaBHjCN (170 mg, 2.70 mmol) was added and the mixture was stirred for a further 4 h at 20 °C, then poured into water. After prolonged cooling the resulting 20 oily precipitate was collected and extracted with CH2CI2. Following filtration the solution was washed with water, dried (Na2S04) and then concentrated under reduced pressure below 30 °C. The residue was chromatographed on silica gel, eluting with C^CVEtOAc (9:2), to provide material that was precipitated from a CH2CI2 solution with petroleum ether at 20 °C to give 5-[2-(tert-25 butyloxycarbonylamino)ethylamino]-1 -(chloromethyl)-3-[(5,6,7-trimethoxyindol-2-yl)carbonyl]-l,2-dihydro-3J?-benz[e]indole (132 mg, 40%), mp 110-115 °C. lH NMR [(CD3)2SO] 5 11.45 (s, 1 H), 8.09 (d, J= 8.5 Hz, 1 H), 7.79 (d, J= 8.3 Hz, 1 H), 7.53-7.26 (underlying v br s, 1 H), 7.49 (t, /= 7.7 Hz, 1 H), 7.33 (t, J= 7.6 Hz, 1 H), 7.04 (s, 1 H), ca 7.07-7.00 (obscured signal, 1 H), 6.97 (s, 1 H), 6.28 (br 30 s, 1 H), 4.68 (t, J= 9.8 Hz, 1 H), 4.45 (dd, J= 11.0,1.4 Hz, 1 H), 4.17-4.07 (m, 1 WO 02/059122 PCT/NZ02/00005 50 H), 3.98 (dd, J= 11.0,3.0 Hz, 1 H), 3.92 (s, 3 H), 3.82 (s, 3 H), 3.80 (s, 3 H), 3.76 (dd, J- 10.7, 8.0 Hz, 1 H), ca 3.3 (br s, obscured by H2O signal but visible after D2O exchange, 2 H), 3.18 (br s, 2 H), 1.39 (s, 9 H). Anal. Calcd. for C32H37CIN4O6: C, 63.1; H, 6.1; N, 9.2; CI, 5.8. Found: C, 63.0; H, 6.1; N, 9.4; CI, 5 5.7%.

A solution of the above compound (122 mg, 0.20 mmol) in dioxane (3 mL) was treated with HCl-saturated EtOAc (3 mL), and the mixture was stood at 20 °C for 1 h. Excess EtOAc was then added to complete separation of the product, which. 10 was collected and recrystallised from MeOH/EtOAc/petroleum ether/HCl to give 5-(2-aminoethylamino)-3-[(5,6,7-trimethoxyindol-2-yl)carbonyl]-l-(chloromethyl)-l,2-dihydro-3iJ-benz[e]indole dihydrochloride 29 (86 mg, 74%), mp >200 °C. JH NMR [ftee base in (CD3)SO] 8 11.46 (br s, 1 H), 8.17 (d, J= 8.6 Hz, 1 H), 7.78 (d, 8.2 Hz, 1 H), ca. 7.5-7.3 (underlying v br s, 1 H), 7.49 (t, J-15 7.6 Hz, 1 H), 7.32 (t, 7.7 Hz, 1 H), 7.04 (s, 1 H), 6.97 (s, 1 H), 6.28 (t, 5.0 Hz, 1 H), 4.67 (t, J= 9.5 Hz, 1 H), 4.45 (dd, J= 11.0,1.3 Hz, 1 H), 4.19-4.07 (m, 1 H), 3.98 (dd, J= 10.9,3.0 Hz, 1 H), 3.92 (s, 3 H), 3.82 (s, 3 H), 3.80 (s, 3 H), 3.77 (dd, J= 11.0, 8.2 Hz, 1 H), 3.12 (br s, 2 H), 2.84 (br s, 2 H). Anal. Calcd. for C27H29CIN4O4.2HCLO.5H2O: C, 54.9; H, 5.5; N, 9.5. Found: C, 55.1; H„ 5.5; N, 20 9.1%.

Example D: Preparation of ancillary ligands l,4,7,10-Tetraazacyclododecane-l,7-dipropanesulfonic acid tetrahydrochloride (32).

A solution of perhydro-3,6,9,12-tetraazacyclopenteno[l,3-/g]acenaphthylene (30) (0.50 g, 2.58 mmol) [Weisman et al., Tetrahedron Lett., 21,1980,335] and 1,3-propanesultone (1.57 g, 12.9 mmol) in CH3CN (20 mL) was stirred at 80 °C under N2 for 72 h. The suspension was cooled to room temperature and the white precipitate was filtered and washed with excess CH3CN to give l,7-bis(3-30 sulfopropyl)-4,10-diaza-l,7-diazomatetracyclo[5.5.2.0.4'14010'I3]tetradecane (31) (0.98 g, 86%): mp 279-281 °C; *HNMR (D20) 8 4.49 (s, 2 H), 3.95 (m, 8 H), 51 3.82 (bd, J= 13.3,2 H), 3.60 (m, 4 H), 3.38 (bd, /= 14.0,2 H), 3.05 (m, 8 H), 2.39 (m, 2 H), 2.27 (m, 2 H); I3C NMR (D20) 8 81.6, 64.2,58.7, 57.6,49.8,49.0, 45.3,21.4. C!6H3oN406S2 requires M+H 439.1685. Found (FAB) 439.1686.

A mixture of 31 (0.50 g, 1.14 mmol) and hydrazine monohydrate (15 mL, 98%) were heated (100 °C) under N2 for 48 h. Excess hydrazine was removed and the residue was dissolved in H20. Acidification with HC1 gave a yellow solution. Evaporation of H20 gave a brown solid (hygroscopic). Trituration with MeOH (x 10) gave 32 (0.59 g, 91%) as a cream powder: mp 322-325 °C; *H NMR (D20) 8 10 3.24 (m, 8 H), 2.94 (m, 12 H), 2.82 (m, 4 H), 1.95 (quintet, /= 7.4,4 H); 13C NMR (D20) 8 53.9,51.7,49.9,45.1,21.7. 1,4,7,10-Tetraazacyclododecane-l-butanesulfonic acid tetrahydrochloride (34).

A solution of 30 (0.50 g, 2.58 mmol) and 1,4-butanesultone (1.75 g, 12.9 mmol) in CH3CN (15 mL) was stirred at 60 °C under N2 for 48 h. The suspension was cooled to room temperature and the white precipitate was filtered and washed with excess CH3CN to give 4-decahydro-4a,6a,8a-triaza-2a-azoniacyclopenta|Xg]acenaphthylen-2a-yl-l-butanesulfonate (33) (0.82 g, 96%): 20 mp 301-303 °C; *H NMR (D20) 8 3.91 (m, 1 H), 3.91 (d, 2.5, 1 H), 3.80 (m 3 H), 3.67 (m, 1 H), 3.58 (d, J= 2.7,1 H), 3.53 (m, 1 H), 3.43 (m, 1 H), 3.24 (m, 4 H), 3.00 (t, J= 13,2 H), 2.87 (m, 5 H), 2.50 (m, 2 H), 2.09 (m, 1 H), 1.98 (m, 1 H), 1.85 (m, 2 H); 13C NMR (D20) 8 86.5,74.4,64.8,60.4,59.6, 53.9, 52.5, 51.1, 50.9, 50.4, 50.3, 46.3,24.2,24.0. Ci4H26N204S requires M+H 331.1804. Found 25 (FAB) 331.1806.

A mixture of33 (0.30 g, 0.92 mmol) and hydrazine monohydrate (6 mL, 98%) was heated (80 QC) under N2 for 36 h. Excess hydrazine was removed and the residue was dissolved in H20. Acidification with HC1 gave a yellow solution. Evaporation of H20 gave a brown solid (hygroscopic). Trituration with MeOH (x 30 10) gave 34 (0.41 g, 97%) as a cream powder: mp 322-325 °C; 'H NMR (D20) 8 52 3.20 (m, 16 H), 2.95 (m, 4 H), 1.78 (m, 4 H); 13C NMR (D20) 8 56.2, 52.9,52.1, 51.6,46.4, 45.5,44.9,25.1,24.4. l,4,7,10-Tetraazacyclododecane-l,7-dipentanoic acid (36).

A solution of 30 (0.10 g, 0.52 mmol) and ethyl 4-iodobutyrate (0.79 g, 3.09 mmol) [Nudelman et al., Bioorg. Chem., 26, 1998,157] in CH3CN (5 mL) was stirred at 60 °C under N2 for 6 days. A further portion of the iodide (0.26 g, 1.03 mmol) was added and the reaction was stirred at 60 °C under N2 for 3 weeks. CH3CN was removed and the residue was partitioned between CH2CI2 and H2O.-10 The aqueous layer was extracted with CH2CI2 (x 6). H20 was evaporated and the residue was solidified with CH3CN/Et20 followed by trituration with Et20 (x 4) to give 1,7-bis(ethoxycarbonylbutyl)-4,10-diaza-1,7- diazoniatetracyclo[5.5.2.0.4'14010'13]tetradecane diiodide (35) (0.32 g, 87%): *H NMR (D2O) 8 4.46 (s, 2 H), 4.17 (q, J= 7.2, 4 H), 3.92 (m, 6 H), 3.77 (nx, 4 H), 15 3.59 (m, 2 H), 3.45 (td,/= 12.8,4.1, 2H), 3.35 (bd,J= 13.9,2H), 3.06(m,4 H), 2.48 (t, J= 7.3, 4 H), 1.96 (m, 2 H), 1.85 (m, 2 H), 1.69 (quintet, 7.3, 4 H), 1.25 (t, /= 7.2,6 H); 13C NMR (D20) 8 178.6, 81.2,64.5, 64.4, 60.2,57.9,49.0, 45.3, 35.9, 24.8,23.8, 16.1. C24H44I2N4O4 requires M+H-I 579.2407. Found (FAB) 579.2410.

A mixture of 35 (0.05 g, 0.08 mmol) and 15% aqueous KOH (5 mL) was stirred at 70 °C under N2 for 48 h. Water was evaporated and the residue was acidified to pH 2.5 with HC1. The mixture was loaded onto aDOWEX 50W-X8 cation exchange resin (H* form). Elution with H2O followed by 0.5M NH3 gave 36 (0.03 25 g, 100%) as a colourless oil: 'H NMR (D20) 8 2.86 (m, 8 H), 2.68 (m, 8H), 2.53 (t,J= 7.4,4 H), 2.20 (q, /= 7.0,4 H), 1.55 (m, 4 H), 1.47 (m, 4 H); 13C NMR (D20) 8 183.1,53.1,49.1,42.8,37.1,24.1,23.8. C18H36N4O4 requires M+H 373.2815. Found (FAB) 373.2810.

Example E. Preparation of metal complexes.

WO 02/059122 PCT/NZ02/00005 53 Preparation of Complex Ml of Table 1. [[Co(cyclen)18a)J(C104)2].

[Co(cyclen)(N02)2](NC>2) (38) [Collman and Schneider, Inorg. Chem. 1966,5, 1380] (1.03 g, 2.79 mmol) was cautiously added with stirring to neat triflic acid (10 mL) cooled in an ice bath. The solution was bubbled with N2 to remove NGX 5 gas and warmed briefly at 40-50 °C until reaction was complete. Dry Et20 (250 mL was added slowly to the above cold solution (ice-bath) with vigorous stirring, and the resulting precipitate was filtered off, washed"(4 x dry Et20) and dried in a desiccator to give [Co(cyclen)(OTf)2](OTf) (39) (1.95 g, 100%). Anal. Calcd. for C11H24C0F9N4011S3: C, 18.49; H, 3.39; N, 7.85. Found: C, 18.43; H, 3.49; N, 10 7.84. HRMS FAB+ [M-OTff calculated for: CioH2oCoF6N406S2= 529.00605. Found: 529.00406. (39) (90 mg, 0.132 mmol) was dissolved in dry CH3CN (3 mL) and 18a (62 mg, 0.132 mmol) was added. To the stirred solution was added iP^NEt (25 mg, 1.5 equiv). This resulted in rapid darkening of the solution to a brown colour but with significant amounts of suspended yellow solid 15 (unreacted/undissolved) 18a present. The mixture was stirred at room temperature for 11 days, during which time nearly all of the suspended solid disappeared. The small amount remaining was removed by filtration through a 0.45 n membrane filter and the filtrate made slightly acidic with dilute aqueous HCIO4. Excess 1 M NaClC>4 (aq) was added and the solution was extracted 4 x with 5 mL CH3NO2. 20 The combined extracts were evaporated to dryness, the residue resuspended in dry Et20 (15 mL) and again evaporated to dryness (first on a Rotovapor, finally on a vacuum line) below 20 °C, to give crude product as brown flakes of glassy material (103 mg, 86%). HRMS FAB [M-CIO4]"1". This material was further purified on reverse-phase HPLC, and the pooled pure fractions were concentrated 25 under reduced pressure, then combined with excess aqueous 1 M NaCl O4 and extracted 5 x with CH2CI2. The combined organic extracts were treated as above to give complex Ml as brownish flakes (-70 mg). HRMS FAB [M-2C104-H]+ Calcd for C&Hii^CICqNtOs; 697.21897. Found, 697.21327. Calcd for C32H4i37ClCoN705; 699.21602. Found, 699.21601.

Preparation of Complex M2 of Table 1. [[Co(cyclen)(18c)](C104)2] 54 [Co(cyclen)(OTfh](OTf) (39) (0.087 g, 0.128 mmol) was dissolved in dry CH3CN (4 mL) and 18c (0.052 g, 0.115 mmol) was added. The mixture was stirred at room temperature for 8 h. then cooled overnight at 5 °C. A small amount of unreacted 18c was removed by filtration and the bright yellow solid washed 5 with cold CH3CN and the washes added to the filtrate. This dark brown solution was reduced to ca. 2 mL by evaporation of solvent under reduced pressure at room temperature and then chromatographed on a short (3.3 x 40 mm) flash silica gel column (0.32-0.60 pm). Elution started with MeOH/CH3NC>2 (5 %) which was stepwise enriched with MeOH up to 15 %. At this concentration the main band . 10 was eluted first followed closely by a small yellow brown band. A stationary red band remains at the top of the column. Removal of the solvent on a rotary evaporator then on a vacuum line to give M2 as a brown glassy residue (0.089 g, 79 %). HRMS FAB+ [M-20Tff Calcd. for Css^s^ClCoNyOs 681.26044. Found, 681.26064; for 37C1 = 683.25749. Found, 683.26086.

Preparation of Complex M3 of Table 1. [[Co(cycIen)(18b)] (004)2] This was prepared as above from 39 (0.101 g, 0.149 mmol) and 18b (0.055 g, 0.118 mmol) to give, after flash chromatography on silica gel, M3 (0.078 g, 67 %). HRMS FAB+ [M-20Tf]+ calculated for CssR^ClCoNgOs 694.25569. Found 20 = 694.25305; for 37C1 = 696.25274. Found = 696.25401.

Preparation of Complex M4 of Table 1. [Cr(acac)2(18a)].

Solid 18a (20 mg, 0.0427 mmol) was added to a solution of [Cr(acac)2(H20)2]C104-2H20 (mixture of cis and trans isomers; Ogino, et al., 25 Inorg. Chem. 1988,27,986) (0.03 g, 0.071 mmol) in dry CH3CN (3 mL). The mixture was stirred and a solution of iP^NEt (6 mg, 0.0464 mmol) in CH3CN (0.5 mL) was added gradually over 1 h. The solution was warmed in an oil bath at 50 °C for 0.5 h, then stirred at ambient temperature for 2 weeks. During this period undissolved 18a gradually disappeared as the complexation reaction proceeded 30 giving a clear red-brown solution. The solvent was removed under reduced pressure and the residue was dissolved in CHC13 (1 -0 mL) and purified by flash 55 chromatography on silica gel. Elution with a CH3CN/CHCI3 gradient firom 0 to 50% CH3CN eluted a single yellow-brown band that trailed somewhat near the bottom of the column. The trailing material was eluted separately with 100% CH3CN. A small amount of green irreversibly absorbed material was left at the 5 top. The main band and tailing fraction were evaporated to dryness under reduced pressure to give yellow-brown powders of Cr(acac)2(18a) (18 mg, 59%) and (5 mg, 16%), respectively.

These two samples gave identical accurate mass spectral results; approximately equal amounts of both [M]+and [M + H]+ions observed with relative intensities consistent with one 35C1 or 37C1 per molecule. FAB+-MS: [M]+ calc. for C34H3535Cl52CrN309= 716.14669. Found, [M]+= 716.14642. [M + H]+ calc. for C34H3637a52CrN309= 719.15157. Found, [M + H]+ = 719.15122. Fragments corresponding to loss of acac ligand are observed, and the base peak corresponds to Cr(acac)2. Analytical HPLC on an RP C-18 column using gradient elution starting from a 1:1 (v/v) mixture of 80% aqueous CH3CN and phosphate buffer (pH = 7.4, 0.04 M) showed one major peak (96.7%) with a prominent UV absorption band at 339 nm. A small amount (0.45%) of uncomplexed 2 could be detected and its identity was confirmed by spiking. Because of the paramagnetic properties of the Cr(III) present in this complex, XH or 13C resonances were not observed by NMR.

Preparation of Complex M5 of Table 1. [Co(Mejdtc)2(18a)] [Co2(Me2dtc)5]BF4 (105 mg, 0.1303 mmol) [Hendrickson et al., J. Chem. Soc. 25 Dalton Trans. 1975,2182] was added to a suspension of 18a (46 mg, 0.0983 mmol) in 5% MeOH/CHaCk (4 mL). iP^NEt (25 mg, 2 equiv) was added to stirred suspension in two portions with the second added one day after the first Stirring was continued at room temperature for 8 days, by which time very little suspended/unreacted 18a was evident, and the colour of the solution was the deep 30 green of the co-product Co(Me2dtc)3. The solution was filtered and the filtrate evaporated under reduced pressure. The residue was taken up in CH2CI2 (2 mL) 56 and chromatographed on a flash silica gel column. Elution began in CH2CI2, and a large green band of Co(Me2dtc)3 was eluted. Stepwise enrichment with CH3CN in increments of 10% was carried out until the product -[Co(Me2dtc)2(18a)] (M5) was eluted (with ca 50% CH3CN/CH2CI2). The main muddy yellow-green band 5 was collected, and solvent was removed under reduced pressure to give the product as a brownish-green amorphous residue (48 mg, 63%). Analytical reverse-phase HPLC indicated no detectable free cytotoxic ligand 18a present.

Preparation of Complex M6 of Table 1. [[Cr(acac)2(29)]ClC>4)] A suspension of 29 (31 mg, 0.058 mmol) in CH3OH (0.5 mL) was treated with a solution of NaOH (5 mg, 0.119 mmol) dissolved in CH3OH (0.5 mL), and the neutralised solution was immediately added to a another containing a mixture of cis- and frans-[Cr(acac)2(OH2)23C104-2H20 (29 mg, 0.069 mmol) dissolved in CH3CN (1.0 mL). The combined mixture was stirred at 50 °C for 15 ™in3 cooled 15 to room temperature and the solvent removed under reduced pressure.

Chromatography on silica gel gave [Cr(acac)2(29)]C104 (M6) as a purple residue after drying under vacuum over silica gel desiccant. HRMS (FAB+/NBA): Calculated [M+J for C37H43N435ClCr08, 758.21834. Found, 758.21745.

Preparation of Complex M7 of Table 1 [[Co(TACN)(8-HQ)(CN)] C104] Co(TACN)(N02)3 was prepared from Na3[Co(N02)6], using the method of Wieghardt et al., Chem. Ber., 1979,112,2220-2230. This was then used to prepare [Co(TACN)(H20)3](OTf)3 (91% yield), essentially by the method of Galsboel et al., Acta Chem. Scand., 1996,50,567-570. [Co(TACN)(H20)3](QTf)3 25 (360 mg, 0.509 mmol) was dissolved in EtOH (9 mL) and 8-hydroxyquinoline (8-HQ) (73 mg, 0.6 mmol) added as a solid. Immediately a solution of Et3N (62 mg) in EtOH (-4 mL) was added to the stirred solution, which was then warmed briefly to complete the coordination of 8-HQ to the cobalt centre. NaCN (150 mg, 4 equiv) was added portionwise, and the mixture was stirred for 24 hours. During 30 the addition of NaCN and occasionally thereafter, the pH was adjusted to ca. 7 by WO 02/059122 PCT/NZ02/00005 57 addition of 0.1 M HCIO4. The red crystals and orange precipitate that formed were dissolved by dilution of the mixture with H2O and the whole was loaded onto a Sephadex SP C-25 cation exchange column and thoroughly washed with H2O. Elution with 0.05 M then 0.1 M NaC104 eluted the major band, and 5 concentration of the eluate by evaporation under reduced pressure produced red-brown crystals of [Co(TACN)(8-HQ)(CN)]C104 (M7) (117 mg, 51%) which were collected and washed with a little ice cold H2O then 3 x with Et20. Anal. Calcd for C16H21N5CIO5C0: C, 41.98; H, 4.62; N, 15.30; CI, 7.74. Found; C, 41.99; H, 4.44; N, 15.28; CI, 7.93.

Biological activity Selected complexes of Table 1, together with the uncomplexed cytotoxic ligands, 15 were evaluated for cytotoxicity (measured as IC50 values in pM following a 4 h aerobic drug exposure) in a panel of mammalian cell lines, and the results are given in Table 2. AA8 is a Chinese hamster ovary line, and the UV4 cell line is a repair-defective ERCC-1 mutant, sensitive to agents whose cytotoxicity is due to bulky DNA adducts. EMT6 is a murine mammary carcinoma line, and SKOV3 is 20 a human ovarian cancer line.

Table 2. Shows the results of the biological activity for various cytotoxins and their metal complexes. IC50 values are mean ± sem (number of experiments in parentheses) for exposure of the indicated cell lines to compounds for 4 hr under 25 aerobic conditions. 58 Table 2 Compound ICso(nM) AA8 UV4 EMT6 SKOV3 Cytotoxic livands 29 0.0058 ± 0.0007 (2) 0.0041 + 0.0003 (2) 0.0028 0.0004 (2) 0.0062 18a 0.00014 ± 0.000022 (7) 0.00007 ± 0.000015 (6) 0.000051 ± 0.000008 (5) 0.00025 ± 0.000037 (8) 27 0.0079 + 0.002 (4) 0.0029 ± 0.006 (4) 0.0026 ± 0.0005 (4) 0.012 ± 0.0023 (4) 8-HQ 2.07 ± 0.02 (3) 2.16 ± 0.12(3) 3.92 ± 1.04(2) 4.07 ± 0.89(2) Ancillary Iwcmds TACN (VHIc: R'-R3=H) 12700± 5770 (2) 10100± 3930 (2) 7710 ± 1010 (2) 13500 ± 5480 (2) Cyclen (IX; Z1-Z4=(CH2y, R1'-R4'=H) 13300 + 2670 (2) 13800± 2180 (2) 9710 ± 2710 (3) 11500± 4410(2) Metal complexes Ml 0.0152 + 0.0006 (2) . 0.0051 ± 0.0002 (2) 0.0133 ± 0.0008 (2) 0.015 ± 0.005 (3) M4 0.088 ± 0.017(3) ' 0.03 ± 0.0018 (3) 0.039 ± 0.011 (3) 0.11 ± 0.018(3) M5 0.028 ± 0.003 (3) 0.015 + 0.001 (3) 0.0095 ± 0.0012 (2) 0.016 ± 0.003 (3) M7 ,670 ± 45.0 (2) 6,140 + 820 (2) 3,580 ± 95.4 (2) 6,380 ± 1750(3) The results of Table 2 show that the cytotoxic ligands 29,18a and 27 are exceptionally cytotoxic. The results of Table 2 also show that metal cpmplexation 59 results in considerable abrogation of cytotoxicity, indicating the utility of this approach in forming less toxic prodrugs of these compounds.

Complex Ml listed in Table 1 was also evaluated for its ability to release the 5 cytotoxic ligands when exposed to ionising radiation in deoxygenated sodium formate buffer (measured as G values in pM/Gy for radiolytic reduction, where the G value for total reductants is 0.68 |iM/Gy), and the results are given in Table 3.

Table 3. G values (jxM/Gy) for release of cytotoxic ligand on radiolytic reduction in deoxygenated sodium formate buffer using 15 |iM prodrug (complex Ml).

No Metal Cytotoxic Ancillaiy ligands G value ligand (jamol/Gy) Ml Co 18a Cyclen 0.75 The results of Table 3 show that certain of these metal complexes also have the 15 potential to cleanly release their cytotoxic ligand in good yield following exposure to ionising radiation. As a specific example, Figure 1 shows the release of cytotoxin 18a (SN 26800) firom complex Ml (SN 27892) when irradiated in 0.1M sodium formate buffer pH 7.0 under hypoxic conditions.

It is thought that the mechanism of activation of the prodrug is as illustrated in the following mechanistic pathway. 60 OMe -iTW OMe IH1: Co1" species (stable, non-toxic) reducing equivalents from ionising radiation or from endogenous reductants V- Co" species (unstable: non-toxic) OMe .p{~VoMe 18a (toxic, stable) The metal complexes also show an ability to be activated by endogenous enzymes under hypoxia, as shown for metal complex Ml in Table 4 and Figure 2. Table 4 5 and Figure 2 also show that the corresponding cytotoxic ligand 18a is not activated by endogenous enzymes under hypoxic conditions. Thus the metal complexes have utility as hypoxia- as well as radiation-activated cytotoxins.

TABLE 4: Activation of metal complex Ml (but not the cytotoxin 18a) tinder 10 hypoxia (4 h exposure). ic50 (nM) A549wt/s SKOV3 WiDr-2 oxic anoxic 1 H< l< oxic anoxic II J- oxic anoxic IUR 18a 0.050 ± 0.016(3) 0.050 ± I 0-79 ^ ' 1 0.014(2) |,|^ , , 0.25 ± 0.037(8) 0.35 ± 0.093(2) \ it ^ Ml .60 ± 0.00(2) 0.38± list 0.16(2) I ±-7.7,(2) J .0 ± 5.0(3) 1.7 ± 0.97(2) 6.6 ± 0.45(2) 1.7 ± 0.10(2) 3.90 x" 0 ^0I2I 61 In Table 4, A549wt/s is a wild-type human colon carcinoma cell line, SKOV3 is a human ovarian cancer cell line and WiDr-2 is a clonal cell line derived ftom the WiDr human colon carcinoma line. ic50s (in jj,M) are determined under both oxic and hypoxic conditions, and the hypoxic cytotoxicity ratio (HCR) is the average 5 inta-experiment ratio of the ic50s measured under oxic and hypoxic conditions.

Wherein the foregoing description reference has been made to reagents, or integers having known equivalents thereof, then those equivalents are herein incorporated as if individually set forth. 0 While this invention has been described with reference to certain embodiments and examples, it is to be appreciated that further modifications and variations may be made to embodiments and examples without departing ftom the spirit or scope of the invention.

Claims (5)

62 10 15 What we claim is: 1 A metal complex represented by Formula I charge qpcountercharge I wherein: A is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci^alkyl, Ci-6alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; B is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci^alkyl, Ci-6alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; D is selected from C or N; E is selected from a direct bond, OH or NR^, where each R1 independently represents H or a Ci_6alkyl optionally substituted with one or more hydroxy or amino groups, when D represents C; M is selected from Co111, Co11, Cr111 or Cr11; Z is selected from O, NR2, where R2 represents H or a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups, S1 and S2 together represent formula V 20 <WrELLECTUAL^RC?EftfyoFRCE! t o JAM £35 WO 02/059122 63 PCT/NZ02/00005 X V wherein X is selected from a group including halogen, CBfe-halogen, CH2OCO-(Ci-C6alkyl optionally substituted with one or more amino or hydroxy groups), 5 CH2-phosphate group or CH2OSO2R3, where R3 represents H or a Ci^alkyl optionally substituted with one or more hydroxy or amino groups, or CH2OSO2NHR4 where R4 represents H or a Ci^alkyl optionally substituted with one or more hydroxy or amino groups; and 0 R is selected from one of formulae VI or VH wherein each Ti, T2 and T3 is independently selected from H, OPOfOIfh, OR5, NR52 or NHCOR5, where each R5 independently represents H, a C^aUcyl optionally substituted with one or more hydroxy or amino groups; or 0(CH2)nNR62, where each n is independently 1,2,3 or 4 and each R6 is independently selected from H or a Ci-galkyl optionally substituted with one or more hydroxy or amino groups and • represents the point of attachment of R to Formula V defined above, and S3 is selected from H, cyano, phosphate, amino, Ci-galkyl, Cj^alkoxy, halogen, C02[(Ci.6allcyl) wherein, said alkyl is optionally substituted with amino, or hydroxy groups]; OR7, NR72, or CONHR7, where each R7 independently represents H, a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups; or S3 represents, an optionally substituted 5 or 6 membered cyclic system optionally containing one or more heteroatoms fused to ring system A VI VII WO 02/059122 64 PCT/NZ02/00005 defined above, wherein said substituents are selected from OH, cyano, phosphate, amino, Ci^alkyl, Ci-gallcoxy, and halogen groups, and wherein ligands LJ-L4 are each independently selected in combinations from 5 anionic monodentate ligands, including CN", SCN", halide, NO2'; bidentate ligands including MeCOCHJCOMe (Jacac; deprotonated in the complex), where J = H, Me, CI, SMe, S02Me, S(CH2)nS03H, S(CH2)»C02H, S(CH2)„0P(0)(0H)2, CH2(CH2)„S03H, CH2(CH2)nC02H, CH2(CH2)„0P(0)(0H)2, S(CH2)nP(0)(0H)2 or CH2(CH2)nP(0)(0H)2, where n is from l-4;or tridentate ligands Vllla-Vinc _(= 10 respectively TACH, TAME and TACN when Ri-R3=H). Villa Vlllb Vlllc wherein each R1-R4 are independently selected from H, Me, CH2(CH2)nS03H, 15 CH2(CH2)„C02H or CH2(CH2)nP(0)(0H) or CH2(CH2)nNR82 where each n is o independently 1, 2, 3 or 4 and each R independently represents H, or a Ci^alkyl optionally substituted with one or more hydroxy or amino groups or L/-L4 can also be selected from any one of the tetradentate ligands IX-XVII, or 20 any two of the bidentate ligands XVHL, or any combination of the bidentate ligands XVHI together with any of the monodentate ligands L'-L4 defined above; WO 02/059122 65 PCT/NZ02/00005 wherein in formulae IX-XVIH, R1 to R8' each independently represent H, Me, 5 CH2(CH2)nS03H, CH2(CH2)„C02H or CH2(CH2)n0P(0)(0H)2 or CH2(CH2)nNMe2, where each n is independently 1,2,3 or 4; each Zl-Z? is independently selected from -(CH2)2-S -(CH2)3-, -CH2OCH2- or -CH2N(R9)CH2-; where R9 represents H, a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups and 10 each Y' is independently selected from H, halogen, S02Me, O(Ci-Cealkyl), NR102, where each R10 is independently selected from H or a Ci-galkyl optionally substituted with one or more hydroxy or amino groups, or Q^CH^nQ2, wherein Q1 is selected from -0-, -CH2-, -NH-, -CONH-, -C02- or -SO2-, and Q2 is selected from -C02H, -SO3H, -0P(0)(0H)2 or -NRll2 where each R11 is 15 independently selected from H or a Ci^alkyl optionally substituted with one or more hydroxy or amino groups; and wherein the overall charge on the complex is neutral, positive or negative and wherein in the case of a non-neutral complex Fcounteroharge is selected from a range of physiologically acceptable-counterions, including halide", N03", NH4+ or Na+; 20 and WO 02/059122 66 PCT/NZ02/00005 wherein q is the required number to neutralise the overall charge on the complex; and including any enantiomeric or diastereomeric form, and any physiologically salt derivative thereof. 5 2 The metal complex according to claim 1 wherein the rings A and B of Formula I as defined in claim 1 together represent an 8-substituted quinoline system. 3 The metal complex according to any claim 1 or claim 2 wherein Z 10 represents -0-. 4 The metal complex according to any claims 1 or 3 wherein Z represents -NH-. 15 5 The metal complex according to any one of claims 1 to 4 wherein R is selected from one of p(CH2)2NMe2 OMe OMe 0(CH2)2NMe2 OMe OMe or OMe OH 20 6 The metal complex according to any one of claims 1 to 5 wherein one of the ligands L^L4 is selected from the following WO 02/059122 67 PCT/NZ02/00005 /—\ , ^NH KL NH HN 7 The metal complex according to any one of claims 1 to 6 wherein X is CH2C1. 5 8 The metal complex according to any one of claims 1 to 7 selected from one of the following; 10 9 A method of providing cancer treatment, which includes the steps of 68 (a) administering to a non-human patient in need of such therapy an effective amount of a metal complex of Formula I as defined in any one of claims 1 to 8, and (a) activating the metal complex of Formula I under hypoxic conditions via reduction, either enzymatically or by a non-enzymatic endogenous reducing agents, or by ionizing radiation, wherein said activation releases a sufficient amount of an effector from said metal complex of Formula I. 10 The method according to claim 9 including the alternative step of activation of the metal complex of Formula I as defined in any one of claims 1 to 8 by radiotherapy radiation. 11 A composition comprising as an active agent a metal complex of Formula I as defined in any one of claims 1 to 8 and a pharmaceutically acceptable excipient, adjuvant or carrier. 12 The use, in the manufacture of a medicament, of an effective amount of a metal complex of Formula I as defined in any one of claims 1 to 8 for use in treating a subject in need of cancer treatment. 13 A metal complex represented by Formula la WO 02/059122 69 PCT/NZ02/00005 charge la wherein: A is selected from a 5 or 6 membered aromatic ring system optionally containing 5 one or more heteroatoms and optionally substituted with one or more Chalky!, Ci^alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; B is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci-galkyl, Ci.6alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; 10 D is selected from C or N; E is selected from a 5 or 6 membered ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci^alkyl, Ci-galkoxy, halogen, hydroxy, phosphate, cyano or amino groups M is selected from Com, Co11, Cr111 or Cr11; 15 Z represents NH2 or NHMe, Q represents H, Ci-galkyl, or (CH^NEfe, when Q represents (CHa^NHb, Q will become a ligand for M and replace one of ligands Lx-L4 defined below, S1 and S2 together represent formula V X o V WO 02/059122 70 PCT/NZ02/00005 wherein X is selected from a group including halogen, CKk-halogen, CH2CN, CH2CC>2-(CrC6alkyl optionally substituted with one or more amino or hydroxy groups), CHrphosphate group, CH2OSO2R3 or OSO2R3 where R3 represents H or 5 a Ci-galkyl optionally substituted with one or more hydroxy or amino groups, or CH2OSO2NHR4 where R4 represents H or a Ci^aliyl optionally substituted with one or more hydroxy or amino groups; and R is selected from one of formulae VI or VH wherein each Ti, T2 and T3 is independently selected from H, OPO(OH)2, OR2, NR22 where each R2 independently represents H, a Ci^alkyl optionally substituted with one or more hydroxy or amino groups or 0(CH2)nNR32, where each n is 15 independently 1,2,3 or 4, and each R3 is independently selected from H or a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups and • represents the point of attachment of Rto Formula V defined above, and S3 is selected from H, cyano, phosphate, amino, Ci^alkyl, Ci-galkoxy, halogen, C02(Ci-6alkyl) wherein said alkyl is optionally substituted with amino, or halogen 20 groups, OR4, NR42, CONHR4, where each R4 independently represents H, a Ci„6alkyl optionally substituted with one or more hydroxy or amino groups; or S3 represents an optionally substituted 4-8 membered cyclic system optionally containing one or more hetoeroatoms fused to ring system A defined above, wherein said substituents are selected from OH, cyano, phosphate, amino, 25 Ci-galkyl, Ci^alkoxy, halogen groups, and VI VII 10 WO 02/059122 71 PCT/NZ02/00005 wherein ligands Ll-LA are each independently selected in combinations from anionic monodentate ligands, including CN", SCN", halide, NO3'; bidentate ligands including MeCOCHJCOMe (Jacac; deprotonated in the complex), where J = H, Me, CI, SMe, S02Me, S(CH2)nS03H, S(CH2)„C02H, S(CH2)n0P(0)(0H)2, 5 CH2(CH2)nS03H, CH2(CH2)nC02H, CH2(CH2)n0P(0)(0H)2, S(CH2)„P(0)(OH)2 CH2(CH2)nP(0)(0H)2, or where each n is independently 1,2,3 or 4; or tridentate ligands VIIIa-VTlIc (respectively TACH, TAME and TACN when Ri-R3=H)s Villa Vlllb Vlllc 10 wherein Rj-ELjare each independently selected firom H, Me, CH2(CH2)nS03H, CH2(CH2)„C02H or CH2(CH2)n0P(0)(0H)2 CH2(CH2)nP(0)(0H)2 or CH2(CH2)nNR52j where each n is independently 1,2, 3 or 4 and each R5 independently represents H, or a Ci-galkyl optionally substituted with one or more 15 hydroxy or amino groups or 20 L'-L4 can also be selected firom any one of the tetradentate ligands IX-XVII, or any two of the bidentate ligands XVIII, or any combination of the bidentate ligands XVIII together with any of the monodentate ligands Ll-L4 defined above; WO 02/059122 72 PCT/NZ02/00005 Me^® R5' ' MeRs' R5\ R8' RVYR" "V\ f" NT rN NT #lkL IV Xr* ^ ^ . R xza xR3' \( \3' R7,/MeMe Rr/iiMe R — r3' IX x XI XII RV6' *\r/y n/ • rnx II II > N=Y rV^NH N \ R1h^NH N-j CN N] L J XNm-> XnhJ j?UVrJt>JV ^ XIV XV XVI XVII wherein in formulae IX-XVIH, R1 to R8 each independently represent H, Me, 5 CH2(CH2)„S03H, CH2(CH2)„C02H or CH2(CH2)n0P(0)(0H)2 or CH2(CH2)nNMe2, where each n is independently 1,2,3 or 4; each Z*-Z4 is independently selected from -(CH2)2-, -(CH2)3-, -CH2OCH2- or -CH2N(R6)CH2-; where R6 represents H, a Ci^alkyl optionally substituted with one or more hydroxy or amino groups and 10 each Y' is independently selected from H, halogen, SO^e, 0(Ci-C6alkyl), NR72, where each R7 is independently selected from H or a Ci.6alkyl optionally substituted with one or more hydroxy or amino groups, or Q1(CH2)nQ23 wherein Q1 is selected from -0-, -CH2-, -NH-, -CONH-, -C02- or -S02-, and Q2 is selected from -C02H, -SO3H, .-0P(0)(0H)2 or-NR82 where each R8 is 15 independently selected from H or a Chalky! optionally substituted with one or more hydroxy or amino groups; and wherein the overall charge on the complex is neutral, positive or negative and wherein in the case of a non-neutral complex Fcountercharge is selected from a range of physiologically acceptable-counterions, including halide", NO3", NH4+ orNa+; 20 and WO 02/059122 73 PCT/NZ02/00005 wherein q is the required number to neutralise the overall charge on the complex; and including any enantiomeric or diastereomeric form, and any physiologically salt derivative thereof. 5 14 Hie metal complex according to claim 13 wherein rings A and B together represent an 8-substituted quinoline system. 15 The metal complex according to claim 13 or claim 14 wherein Z represents -OH. 16 The metal complex according to claim 13 or claim 14 wherein Z represents -NH. 17 The metal complex according to any one of claims 13 to 16 wherein R is 15 selected from one of 10 OMe H 20 18 The metal complex according to any one of claims 13 to 17 wherein one of the ligands LJ-L4 is selected ftom the following 74 NH N N HN /—\ 0 S-OH II o y or Q OH HO' 0 19 The metal complex according to any one of claims 13 to 18 wherein X is CH2C1. 20 The metal complex according to any one of claims 13 to 17 and 19 which represents 21 A method of providing cancer treatment, which includes the steps of (a) administering to a non-human patient in need of such therapy an effective amount of a metal complex of Formula la as defined in any one of claims 13 to 20, and (b) activating the compound of Formula la under hypoxic conditions via reduction, either enzymatically or by non-enzymatic endogenous reducing agents, or by ionising radiation, + OMe >*K J Me O VN WO 02/059122 75 PCT/NZ02/00005 wherein said activation releases a sufficient amount of an effector, from said effective amount of the compound of Formula la. 22 The method according to claim 21 including the alternative step of 5 activation of the metal complex of Formula la as defined in any one of claims 13 to 20 by radiotherapy radiation. 23 A composition comprising as an active agent a metal complex of Formula la as defined in any one of claims 13 to 20 and a pharmaceutically acceptable 10 excipient, adjuvant or carrier. 24 The use, in the manufacture of a medicament, of an effective amount of a compound of Formula la as defined in any one of claims 13 to 20 for use in treating a subject in need of cancer treatment. 15 25 A heterocyclic compound of Formula XIX. XIX wherein 20 A is selected from a 5 or 6 membered ring system optionally containing one or more additional heteroatoms and optionally substituted with one or more Cj-galkyl, Ci-6alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; B is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more C^aUcyl, 25 Ci-salkoxy, halogen, hydroxy, phosphate, cyano or amino groups; WO 02/059122 76 PCT/NZ02/00005 Z is selected from OH or NR^, where each R1 independently represents H or Cj-Ceallcyl optionally substituted with one or more amino, hydroxy, a halogen or cyano groups; 5 S1 and S2 together represent formula V wherein X is selected from a leaving group including halogen, CHrhalogen, CH2CN, CH2CC>2-(Ci-C6alkyl optionally substituted with one or more amino or 10 hydroxy groups), CH2-phosphate group, CH20S02R3 where R3 represents H or a Ci-galkyl optionally substituted with one or more hydroxy or amino groups, or CH2OSO2NHR4 where R4 represents H or a C^aHcyl optionally substituted with one or more hydroxy or amino groups; and R is selected from one of formulae VI or VII wherein each Ti, T2 and T3 is independently selected from H, OPO(OH)2, OR5, NR52 where each R5 independently represents H, a Ci.galkyl optionally substituted 20 with one or more hydroxy or amino groups or 0(CH2)nNR62, where each n is independently 1,2,3 or 4 and each R6 is independently selected from H or a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups; • represents the point of attachment to Formula XIX defined above; S3 is selected from H, cyano, phosphate, amino, Ci^alkyl, Ci-6alkoxy, halogen, 25 C02[(Ci-6allcyl) wherein said alkyl is optionally substituted with amino, or X o V 15 VI WO 02/059X22 77 PCT/NZ02/00G05 hydros groups], OR7, NR72, CONHR7 where each R7 independently represents H, a C^aUsy! optionally substituted with, one or more hydroxy or amino groups; or S3 represents an optionally substituted 4-8 membered cyclic system optionally containing one or more hetoeroatoms fused to ring system A defined above, wherein said substituents are selected from OH, cyano, phosphate, amino, Ci^alkyl, Ci.6alkoxy, and halogen groups; and including any enantiomeric or diastereomeric form, and any physiologically salt derivative thereof, with the proviso that when Z, A, B, X, S1, S2 and S3 together represent R does not represent one of the following WO 02/059122 78 PCT/NZ02/00005 26 The heterocyclic compound according to claim 25 wherein the rings A and B together represent an 8-substituted quinoline system. 27 The heterocycUc compound according to claim 25 or claim 26 wherein Z 5 represents -OH. 28 The heterocyclic compound according to any one of claims 25 to 27 wherein Z represents -NH2. 10 29 The heterocyclic compound according to any one of claims 25 to 28 wherein R is selected from one of 30 The heterocyclic compound according to any one of claims 25 to 29 15 wherein X is-CH2C1. 31 The heterocyclic compound according to any one of claims 25 to 30 selected ftom one of the following 20 l-(chloromethyl)-5-hydroxy-3-[(5,6,7-trimethoxyindol-2-yl)carbonyI]-2,3-dihydro-liy-pyrrolo[3,2-^quinoline, OMe H l-(chloromethyl)-3-({5-[2-(dimethylamino)ethoxy]-indol-2-yl}carbonyl)-5-hydroxy-253-dihydro-l#-pyrrolo[3,2-/]quinolme, 79 1 -(chloromethyl)-3-((2E)-3 - {4- [2-(dimethylamino)ethoxy]phenyl} -2-propenoyl)-5 -hydroxy-2,3 -dihydro-1 if-pyrrolo [3,2f] quinoline, 1 -(chloromethyl)-5 -hydroxy-3 - [(5-methoxyindol-2-yl)carbonyl] -2,3 -dihydro-1H-pyrrolo [3,2f\ quinoline, 1 -(chloromethyl)-5-hydroxy-3 - [(2£)-3 -(4-methoxyphenyl)-2-propenoyl] -2,3 -dihydro-1 //-pyrrolo [3,2/] quinoline, 1 -(chloromethyl)-5-hydroxy-3 - [(2£)-3 -(3 -hydroxy-4-methoxyphenyl)-2-propenoy 1] -2,3 -dihydro-1 //-pyrrolo [3,2/] quinoline and, 5 -amino-1 -(chloromethyl)-3 - [(5,6,7-trimethoxyindol-2-yl)carbonyl] -2,3 -dihydro-iif-pyrrolo[3,2/]quinoline. 32 A method of providing cancer treatment, which includes the step of administering to a non-human patient in need of such therapy an effective amount of a heterocyclic compound of Formula XIX, as defined in any of claims 25-31, XIX wherein A is selected from a 5 or 6 membered ring system optionally containing one or more additional heteroatoms and optionally substituted with one or more WO 02/059122 80 PCT/NZ02/00005 Ci-6alkyl, C^alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; B is selected ftom a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci^alkyl, Ci-6alkoxy, halogen, hydroxy, phosphate, cyano or ainino groups; 5 Z is selected from OH or NR^, where each R1 independently represents H or Ci-Cealkyl optionally substituted with one or more amino, hydroxy, a halogen or cyano groups; S1 and S2 together represent formula V ^ O V 10 wherein X is selected from a leaving group including halogen, CH2-halogen, CH2CN, GEtrphosphate group, CH2CO2R2, where R2 represents Ci-C6alkyl optionally substituted with one or more amino or hydroxy groups; CH20S02R3 where R3 represents H or a Cj^alkyl optionally substituted with one or more hydroxy or amino groups, or CH20S02NHR4 where R4 represents H or a Ci-6alkyl 15 optionally substituted with one or more hydrogen or amino groups; and R is selected from one of formulae VI or VII 20 wherein each Ti, T2 and T3 is independently selected from H, OPO(OH)2, OR5, NR52 where each R3 independently represents H, a C^alkyl optionally substituted with one or more hydroxy or amino groups or 0(CH2)nNR62, where each n is independently 1,2,3 or 4 and each R6 is independently selected from H or a C1-6 alkyl optionally substituted with one or more hydroxy or amino groups; 25 • represents the point of attachment to Formula XIX defined above; WO 02/059122 81 PCT/NZ02/00005 10 S3 is selected from H, cyano, phosphate, amino, Ci^alkyl, Ci^alkoxy, halogen, C02[(Cj-6aIkyl) wherein said alkyl is optionally substituted with amino or hydroxy groups], OR7, NR72, CONHR7 where each R7 independently represents H, a Ci-galkyl optionally substituted with one or more hydroxy or amino groups; or S3 represents an optionally substituted 4-8 membered cyclic system optionally containing one or more hetoeroatoms fused to ring system A defined above, wherein said substituents are selected from OH, cyano, phosphate, amino, Ci-galkyl, Ci-ealkoxy, and halogen group; and including any enantiomeric or diastereomeric form, and any physiologically salt derivative thereof. 33 The method according to claim 32 wherein the rings A and B of Formula XIX together represent an 8-substituted quinoline system. 34 The method according to claim 32 or claim 33 wherein Z of Formula XIX 15 represents-OH. 35 The method according to any one of claims 32 to 34 wherein Z of Formula XEX represents -NH2. 20 36 The method according to any one of claims 32 to 35 wherein R of Formula XIX is selected from one of OMe p(CH2)2NMe2 / /=/ ^5^0(CH2)2NMe2 XM OMe H OMe OMe /==( ^.OMe .^.OMe OH jrjj t °r WO 02/059122 82 PCT/NZ02/00005 37 The method according to any one of claims 32 to 36 wherein X of Formula XIX is -CEfeCl. 38 The method according to any one of claims 32 to 37 wherein Formula XIX 5 is selected firom one of the following 1 -(chloromethyl)-5 -hydroxy-3 - [(5,6,7 -trimethoxyindol-2-yl)carbonyl] -2,3 -dihydro-1 if-pyrrolo [3,2-/[ quinoline, 10 1 -(chloromethyl)-3 -( { 5-[2-(dimethylamino)ethoxy]-indol-2-yl} carbonyl)-5-hydroxy-2,3-dihydro-lB-pyrrolo[3,2-/jquinolme, l-(chloromethyl)-3-((2£Ti-3-{4-[2-(dimethylamino)ethoxy]phenyl}-2-propenoyl)-5-hydroxy-2,3-dihydro-lif-pyrrolo[3,2^quinoline} 15 . 1 -(chloromethyl)-5 -hydroxy-3 - [(5 -methoxymdol-2-yl)carbonyl] -2,3-dihydro-1H-pyrrolo[352-/]quinoline, 1 -(chloromethyl)-5 -hydroxy-3 - [(2E)-3 -(4-methoxyphenyl)-2-propenoyl] -2,3 -20 dihydro-1 i^-pyrr olo [3,2-f\quinoline, l-(chloromethyl)-5-hydroxy-3-[(2£)-3-(3-hydroxy-4-methoxyphenyl)-2-propenoylj-2,3 -dihydro- liJ-pynrolo[3,2-/}quinoline and, 25 5-amino-l-(chloromethyl)-3-[(5,6,7-trimethoxyindol-2-yl)carbonyl]-2,3-dihydro-iff-pytrolo[3^-/( quinoline. 39 A composition comprising as an active agent a compound of Formula XIX WO 02/059122 83 PCT/NZ02/00005 S z XIX wherein A is selected from a 5 or 6 membered ring system optionally containing one or 5 more additional heteroatoms and optionally substituted with one or more C^aUcyl, Ci-ealkoxy, halogen, hydroxy, phosphate, cyano or amino groups; B is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci^alkyl, C^alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; 10 Z is selected from OH or NR^, where each R1 independently represents H or Cr C6alkyl optionally substituted with one or more amino, hydroxy, a halogen or cyano groups; Z is selected from O or NR1, where R1 represents H or Ci-Cgalkyl optionally 15 substituted with one or more amino, hydroxy, a halogen or cyano groups; S1 and S2 together represent formula V wherein X is selected from a leaving group including halogen, GHh-halogen, 20 CTfeCN, CHrphosphate group, CH2C02R2, where R2 represents Ci-C6allcyl optionally substituted with one or more amino or hydroxy groups; CH2OSO2R3 where R3 represents H or a Ci^alkyl optionally substituted with one or more hydroxy or amino groups, or CH2OSO2NHR4 where R4 represents H or a Ci^alkyl optionally substituted with one or more hydroxy or amino groups; and X o V WO 02/059122 84 PCT/NZ02/00005 R is selected from one of formulae VI or VII 5 wherein each Ti, T2 and T3 is independently selected from H, OPO(OH)2, OR5, NR52 where each R5 independently represents H, a Ci-galkyl optionally substituted with one or more hydroxy or amino groups or 0(CH2)nNR62, where each n is independently 1,2,3 or 4 and each R6 is independently selected from H or a Ci-gallcyl optionally substituted with one or more hydroxy or amino groups; 10 • represents the point of attachment to Formula XIX defined above; S3 is selected from H, OH, cyano, phosphate, amino, Ci-6alkyl, Ci^alkoxy, halogen, C02[(Ci-6allcyl) wherein said alkyl is optionally substituted with amino, or hydroxy groups], OR7, NR7, CONHR7 where each R7 independently represents H, a Ci.6alkyl optionally substituted with one or more hydroxy or amino groups; 15 or S3 represents an optionally substituted 4-8 membered cyclic system optionally containing one or more hetoeroatoms fused to ring system A defined above, wherein said substituents are selected from OH, cyano, phosphate, amino, Ci^alkyl, Ci^allcoxy, and halogen groups; and including any enantiomeric or diastereomeric form, and any physiologically salt derivative thereof and 20 a pharmaceutically acceptable excipient, adjuvant or carrier. 40 The composition according to claim 39 wherein the rings A and B of Formula XIX together represent an 8-substituted quinoline system. 25 41 The composition according to claim 39 or claim 40 wherein Z of Formula XIX represents -OH. WO 02/059122 85 PCT/NZ02/00005 42 The composition according to any one of claims 39 to 41 wherein Z of Formula XIX represents -NH2. 43 The composition according to any one of claims 39 to 42 wherein R of Formula XIX is selected from one of 0(CH2)2NMe2 - 0(CH2)2NMe2 OMe OMe or OMe OH 44 The composition according to any one of claims 39 to 43 wherein X of 10 Formula XIX is -CH2C1. 45 The composition according to any one of claims 39 to 44 wherein Formula XIX represents one of the following 15 l-(chloromethyl)-5-hydroxy-3-(5,6,7-trimethoxyindol-2-ylcarbonyl)-2,3-dihydro-1 i^-pyrrolo [3,2-f[ quinoline, l-(chloromethyl)-3-({5-[2-(dimethylamino)ethoxy]-l#-indol-2-yl}carbonyl)-5-hydroxy-2,3-dihydro4ff-pyrrolo[3,2^quinoline, 20 l-(chloromethyl)-3-((25)-3-{4-[2-(dimethylamino)ethoxy]phenyl}-2-propenoyl)-5-hydroxy-2s3-dihydro-l^T-pyrrolo[3,2-^[ quinoline, WO 02/059122 86 PCT/NZ02/00005 l-(chloromethyl)-5-hydroxy-3-[(5-methoxy4#-indol-2-yl)carbonyl]-2,3-dih.ydro-l#-pyrrolo[3;2-j]quinoline, l-(chloromethyl)-5-hydi*oxy-3-[(2iJ)-3-(4-methoxyplienyl)-2-propenoyl]-2,3- 5 dihydro-1 if-pyrrolo[3,2-J] quinoline,

1 -(chloromethyl)-5-hydroxy-3 - [(2E)-3 -(3 -hydroxy-4-methoxyphenyl)-

2-propenoyl]-2,

3 -dihydro-1 i?-pyrrolo [3,2-/]quinoline, and 10 5-amino-1 -(chloromethyl)-3-[(5,6,7-trimethoxyindol-2-yl)carbonyl]-2,3-dihydro-iH-pyrrolo[3,2-/]quinoline. 46 The use, in the manufacture of a medicament, of an effective amount of a compound of Formula XIX wherein A is selected from a 5 or 6 membered ring system optionally containing one or more additional heteroatoms and optionally substituted with one or more 20 Ci-ealkyl, Ci^alkoxy, halogen, hydroxy, phosphate, cyano or amino groups; B is selected from a 5 or 6 membered aromatic ring system optionally containing one or more heteroatoms and optionally substituted with one or more Ci^alkyl, Ci-ealkoxy, halogen, hydroxy, phosphate, cyano or amino groups; Z is selected from OH or NR^, where each R1 independently represents H or Ci-25 Cealkyl optionally substituted with one or more amino, hydroxy, a halogen or cyano groups; S 15 Z XIX WO 02/059122 87 PCT/NZ02/00005 V wherein X is selected from a leaving group including halogen, CHfe-halogen, CH2CN, CH2-phosphate group, CH2CO2R2, whereR2 represents Ci-C6alkyl 5 optionally substituted with one or more amino or hydroxy groups; CH2OSO2R3 where R3 represents H or a Ci.6alkyl optionally substituted with one or more hydroxy or amino groups, or CH2OSO2NHR5 where Rs represents H or a Ci^alfcyl optionally substituted with one or more hydroxy or amino groups; and R is selected from one of formulae VI or VII wherein each Ti, T2 and T3 is independently selected from H, OPO(OH)2, OR5, NR52 where each R5 independently represents H, a Cj^alkyl optionally substituted 15 with one or more hydroxy or amino groups or 0(CH2)nNR62, where each n is Ci-6allcyl optionally substituted with one or more hydroxy or amino groups; • represents the point of attachment to Formula XIX defined above; S3 is selected from H, OH, cyano, phosphate, amino, Ci^alkyl, Ci-gallcoxy, 20 halogen, C02[(Ci_6alkyl) wherein said alkyl is optionally substituted with amino, or hydroxy groups], OR7, NR7, CONHR7 where each R7 independently represents H, a Ci-galkyl optionally substituted with one or more hydroxy or amino groups; or S3 represents an optionally substituted

4-8 membered cyclic system optionally 10 VI VII independently 1,2,3 or 4, and each R6 is independently selected from H or a WO 02/059122 88 PCT/NZ02/00005 containing one or more hetoeroatoms fused to ring system A defined above, wherein said substituents are selected from OH, cyano, phosphate, amino, Ci.6alkyl, Ci-6alkoxy, and halogen groups, and including any enantiomeric or diastereomeric form, and any physiologically salt derivative thereof, 5 for use in treating a subject in need of cancer treatment 47 The use according to claim 46 wherein the rings A and B of Formula XIX together represent an 8-substituted quinoline system. 10 48 The use according to claim 46 or claim 47 wherein Z of Formula XIX represents -OH. 49 The use according to any one of claims 46 to 48 wherein Z of Formula XIX represents -NH2. 15 50 The composition according to any one of claims 46 to 49 wherein R of Formula XIX is selected from one of OMe p(CH2)2NMe2 V /=V r^.O{CH2)2NMe2 lif* fV /vAJ ^NfbMe ' "HI ' OMe /_/ ^.OMe ^.OMe OH f or 20 51 The composition according to any one of claims 46 to 50 wherein X of Formula XIX is-CH2CI. 52 The composition according to any one of claims 46 to 51 wherein Formula XIX represents one of the following WO 02/059122 89 PCT/NZ02/00005 l-(chloromethyl)-

5-hydroxy-3-(5,6,7-trimethoxyindol-2-ylcai-bonyl)-2,3-dihydro-lff-pyrrolo [3,2-/lquinoliiie, 5 l-(cHoromethyl)-5-hydioxy-({5-[2-(dimethylamino)ethoxy]-l#-indol-2-yl}carbonyl)-2,3-dihydro-liJ-pyrrolo[3,2^quittoline, l-(chloromethyl)-3-((2£)-3-{4-[2-(dimethylamino)ethoxy]phenyl}-5-hydroxy-2-propenoyl)-2,3-dihydro-1 F-pyrrolo [3,2-./]quinoline, 10 l-(chloromethyl)-5~hydroxy-3-[(5-methoxy-li7-indol-2-yl)carbonyl]-2.,3-dihydro-lif-pyrrolo[3,2^]quinoline, l-(chloromethyl)-5-hydroxy-3-[(2£T)-3-(4-methoxyph.enyl)-2-propenoyl]-2,3-15 dihydro-1 if-pyrrolo [3,2-/jquinoline, l-(chloromethyl)-3-[(2£)-3-(3-hydroxy-4-methoxyphenyl)-2-propenoyl]-2,3- dihydro-l#-pyrrolo[3,2-/]quinolin-5-ol, and 20 5-amino-l-(chloromethyl)-3-[(5,6,7-trimethoxyindol-2-yl)carbonyl]-2,3-dihydro-1 ff-pyrrolo [3,2-/jquinoline. enantiomeric or diastereomeric forms, or any mixtures of such forms, and also any physiologically functional salt derivatives thereof. 25 53 A method of preparing a metal complex according to any one of claims 1 to 8 or 13 to 20, including the step of coupling a heterocyclic compound defined in any one of claims 25 to 31 with one or more of ligands L'-L4, wherein ligands Ll-L4 are each independently selected in combinations from anionic 30 monodentate ligands, including CN", SCN", halide, NO3"; bidentate ligands including MeCOCHJCOMe (Jacac; deprotonated in the complex), where J = H, WO 02/059122 90 PCT/NZ02/00005 Me, CI, SMe, S02Me, S(CH2)nS03H, S(CH2)nC02H, S(CH2)n0P(0)(0H)2, CH2(CH2)nS03H, CH2(CH2)nC02H, S(CH2)„P(0)(0H)2 or CH2(CH2)nP(0)(0H)2, where each n is independently 1,2,3 or 4; or tridentate ligands Vllla-VIHc (respectively TACH, TAME and TACN when Ri-R3=H), 5 wherein R1-r4 are each independently selected ftom H, Me, CH2(CH2)nS03H, CH2(CH2)nC02H or CH2(CH2)„0P(0)(0H)2 CH2(CH2)nP(0)(0H)2 or 10 CH2(CH2)ftlSrR52) where each n is independently 1,2,3 or 4 and each R5 independently represents H, or a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups or LJ-L4 can also be selected from any one of the tetradentate ligands IX-XVTI, or 15 any two of the bidentate ligands XVIII, or any combination of the bidentate ligands XVUI together with any of the monodentate ligands L1-!,4 defined above; Villa Vlllb Vlllc IX X XI XII XIII R V XIV XV XVI XVII XVIll WO 02/059122 91 PCT/NZ02/00005 wherein in formulae IX-XVIH, R1 to R8' each independently represent H, Me, CH2(CH2)nS03H, CH2(eH2)nC02H or CH2(CH2)nOP<P)(OH)2 or CH2(CH2)nNMe2, where each n is independently 1,2,3 or 4; each Z'-Z4 is independently selected from -(CH2)2-, -(CHhV, -CH2OCH2- or -5 CH2N(R6)CH2-; where R6 represents H, a Ci^alkyl optionally substituted with one or more hydroxy or amino groups and each Y' is independently selected from H, halogen, S02Me, 0(Ci-C6alkyl), NR72, where each R7 is independently selected from H or a Ci-6alkyl optionally substituted with one or more hydroxy or amino groups, or Q^CH^nQ2, wherein 10 Q1 is selected from -0-, -CH2-, -NH-, -CONH-, -C02- or -S02-, and Q2 is selected from -C02H, -SO3H, -0P(0)(0H>2 or-NR82 where each R8 is independently selected from H or a Ci^alkyl optionally substituted with one or more hydroxy or amino groups, and wherein the ligands are complexed with a metal selected from Com, Co11, Cr111 or Cr11. 54 A method of preparing a hetero'cylic compound as defined in any one of claims 25 to 32, including the following reaction pathway 15 R 20 55 The method according to claim 54 including the further steps represented by the pathway WO 02/059122 92 PCT/NZ02/00005 wherein OH 18a-f o c: R = d: R = 0(CH2)2NMe2 OMe e: R = f. R OMe and 56 A metal complex represented by Formula I substantially as herein described with reference to the Examples thereof. 57 A metal complex of Formula I according to any one of claims 1 to 8 substantially as herein described with reference to the Example thereof. 58 A method according to any one of claims 9 or 10 substantially as herein described with reference to the Examples thereof. 59 A composition according to claim 11 substantially as herein described with reference to the Examples thereof. 60 A metal complex represented by Formula la substantially as herein described with reference to the Examples thereof. INTELLECTUAL PROPERTY OFFICE OF N.Z. 2 t JUL 2003 BPcru/cn 93 61 A metal complex of Formula la according to any one of claims 13 to 20 substantially as herein described with reference to the Examples thereof. 62 A method according to claim 21 or claim 22 substantially as herein described with reference to the Examples thereof. 63 A composition according to claims 23 substantially as herein described with reference to the Examples thereof. 64 A compound of Formula XIX substantially as herein described with reference to the Examples thereof. 65 A compound of Formula XIX as claimed in any one of claims 25 to 31 substantially as herein described with reference to the Examples thereof. 66 A method according to any one of claims 32 to 3 8 substantially as herein described with reference to the Examples thereof. 67 A composition according to any one of claims 39 to 45 substantially as herein described with reference to the Examples thereof. 68 A method according to any one of claims 53 to 55 substantially as herein described with reference to the Preparative Examples and Examples thereof. END OF CLAIMS INTELLECTUAL PROPERTY OFFICE OF N.Z. 2 1 JUL 2003 RECEIVED