AU2002227786B2 - Synthesis for the preparation of compounds for screening as potential tubulin binding agents - Google Patents

Description

P c0P0c6412,6: 122fA 160 201 I/ 124X) O-1o Synthesis for the Preparation of Compounds for Screening as Potential Tubulin Binding Agents.

FIELD OF THE INVENTION oO The present invention relates generally to chemical compounds and methods for 00 their preparation. In particular, the invention relates to chemical compounds, and libraries thereof, which may possess useful therapeutic activity, and their use in methods of therapy 8,as well as compositions containing said compounds.

BACKGROUND OF THE INVENTION The search for effective chemotherapeutic drugs relies on the discovery and development of chemical compounds which possess biologically potent anti-tumour activities. While many naturally occurring compounds have been recognised as possessing this desirable activity, eg taxol, the inherent difficulties which may be associated with the isolation and purification of naturally occurring small quantities have prompted extensive efforts directed towards the chemical synthesis of analogues of these bioactive compounds and other potentially bioactive molecules.

Compound libraries prepared by combinatorial methods now play an increasing role in the discovery of bioactive molecules. These libraries can be subjected to high throughput screening methods which allow for the rapid identification of potential new drug candidates. More recently, such libraries have also found utility in molecular biology as an aid to understanding various biological pathways. To reduce the cost of drug discovery using these methods, efficient means of producing molecular diversity need to be identified. Traditionally, efficiency in synthesis has been defined as providing the maximum yield of a targeted product. In diversity-orientated synthesis, efficiency is redefined as providing the maximum range of different structural entities from the minimum number of starting components.

One class of compounds which have attracted attention are those which inhibit tubulin assembly and prevent its polymerization into microtubules. Compounds with P opcrlUtrnm IX6I 226416)2 I doc-2i/1 (1/2V1 -2- U tubulin binding activity are thus effective anti-mitotic agents and include colchicine, Svinblastine, vincristine and taxol. Another tubulin polymerization inhibitor which C has attracted recent interest is Combretastatin A4 which is a powerful inhibitor of tubulin polymerization with an ICso value of -2-3 M and which has been shown to I display potent and selective toxicity toward tumour vasculature. Its 3'-hydroxy disodium phosphate ester is currently the subject of clinical trial.

(N

0However, despite its promising activity, the compound readily isomerises to its 0C inactive (E)-isomer and efforts have been directed towards the identification of configurationally stable analogues. Recently, independent researchers have identified compounds (US Patent No. 5,886,025) and (Madarde, M. el al Bioorg, Med. Chem Lett., 1999, 2303) as showing moderate tubulin binding and anti-mitotic activity.

However, in contrast, the benzofuran did not exhibit tubulin binding activity (Banwell, et al Aust J. Chem., 1999, 52, 767-774).

OMe MeO OMe OMe OMe o m e MeO

(B)

OMe OMe MeO OMe MeO OMe O e O H MeO 0 e /IT V-OMe P op a IU(C 12 264 1w 20)t I do-2 UN1 N I 0 -3- U Accordingly, there exists a need for new synthetic methods which can allow for the Sgeneration of libraries of compounds which can be screened for desired bioactivities, such C as tubulin binding and anti-mitotic activity, and for the discovery of new compounds which possess this activity.

00 It has now been found that a number of compounds prepared by new synthetic methods have been found to possess useful bioactivity. These compounds can be variously Sprepared from a small group of starting substrates to provide a combinatorial library of C compounds which can be screened for bioactivity.

SUMMARY OF THE INVENTION Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this specification relates.

The present invention provides new methods for generating libraries of compounds including benzothiophenes, benzofurans, indoles, indanones and indenones, as well as nonbenzo-fused analogues, for biological screening. The invention further provides new compounds which exhibit tubulin binding anti-mitotic activity, and processes for their preparation.

In a first aspect, there is provided a combinatorial library of 2 or more chemical compounds each compound comprising the reaction product derived from at least two substrates selected from and P 64p.~n64,I\26'I 264 1 o) dMc2 I/I 121 -4- 0

ON

o (a) RlA RIA 0 R o Hal (ii) R B o Hal o or SR1D

RID

O

O

(b) R2-- or a metallated form thereof; or (ii) R2 Hal; or (iii) R1A PsS R1 R 2 Ric (c) 3 L or a metallated form thereof wherein L is replaced by a metal; or (ii) Hal; wherein RIA-RID are independently selected from hydrogen, hydroxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted acyloxy, amino, optionally substituted alkylamino, optionally substituted dialkylamino or any 2 adjacent RIA-RID together form

-O-CH

2 P "crpi\Ial2QX)61\ 22641( 20 11 doc.2 lr 112t(lM 0 0 SHal is I, Br or Cl; X' is OH, SPs (wherein Ps is a sulfur-protecting group capable of stabilising a positive 0 N charge), NPN (wherein PN is a nitrogen-protecting group), or NHR (wherein R is sulfonyl, trifluoroacyl, Ci.7acyl, Ci.

6 alkyl, C2- 6 alkenyl, C2- 6 alkynyl, or an aryl or O heteroaryl group);

R

2 and R 3 are independently optionally substituted aryl or optionally substituted heteroaryl Ni groups; 0L is a leaving group.

In another aspect, the invention provides for combinational library of compounds for screening, as potential tubulin polymerisation inhibitors, said library comprising two or more compounds of formulae to said compounds being the reaction products of the following substrates as previously defined: and to produce compounds of formulae and (F) R3 R1A R3 R1A 0 X

X

R

R2 R2

R

1

R

RD R1D (F) and to produce compounds of formula and to produce compounds of formulae or (K) P %4puV a I.1IA)2264164) 211 dM-21 I If216 I-6- RlB RlAB 00 RiR RID(G) RD(H) l1A l RIB

R

3

RBR

RiD RD(K

RIB

RIC xR Rl(IM and to produce compounds of formulae and (K) and to produce compounds of formula (L) P Wpu~n a WNMMlI2264164) 20)11 doc.21/1I If21l1 -7and to produce compounds of formula (M) I R 2

(M

RID

(b)(iii) and to produce compounds of formula (P) RI

R

2 (p) (b)(iii) and to produce compounds of formula (Q) 0 R1B' R2 (Q) wherein R 2 R3, RIA RID, and are as defined above, and X=O, S, or NR (wherein R is H, sulfonyl, CI.

6 alkyl, C 2 6 alkenyl, C 2 6 alkynyl, C 1 7 acyl, or an aryl or heteroaryl group).

In a further aspect the present invention provides a combinational library of intermediates useful for the preparation of compounds of formulae said intermediates being the reaction products of the following substrates; P %oer W 2Z6MI2641 2NI dmc. Ill112(X)6 -8and to produce intermediates of formula for use in preparing N- compounds of formulae and (F)

RIBA

CIR,

0

XP

P %opcrbnIgIKW12264 i) 20) .1 dO.2 1,11 ro(r, -9and to produce intermediates of formulae and for use in preparing compounds of formulae or 00 RlD (F) R1A 0 RjB -Hal

R

1 D P \Opnlmll(lHI 221,41 2011 dc2ll l 121l6 N 0 o and to produce intermediates of formulae and for use IND in preparing compounds of formula or (M)

C

R 1 A O ROA o RIBRIB

MY

SR

2

R

1 c H RC H -R2 RiD Rio (b)(iii) with itself to produce intermediates of formulae S '-R2 for use in preparing compounds of formulae and wherein RIA-RID, R 2

R

3 X, Hal, and are as defined above, P is a protection group and MY is Sn(alkyl) 3 or B(OR) 2 wherein R is H, alkyl, alkenyl or alkynyl.

In another aspect, the invention provides a combinatorial library of at least two compounds of formula R1A

R

3 RiB

A'

P %Opcr.InaU)6\I 204 IN 1 2011 co(2 1Il If2tx

O

wherein N Xis O; A and A' are independently selected from C=O,or a single bond; 00 is a single or double bond.

RIA- RID are independently selected from hydrogen, hydroxy, optionally substituted C alkoxy, optionally substituted alkyl, optionally substituted alkenyl, optionally 0 substituted alkynyl, optionally substituted acyloxy, amino, optionally substituted C alkylarnino, optionally substituted dialkylamino, optionally substituted acylamino or any 2 adjacent RIA-RID together form -O-CH 2

R

2 and R 3 are independently optionally substituted aryl or optionally substituted heteroaryl groups.

In a further aspect the invention provides a compound of formula

R

3

C

R3B R3E R3A

A'

S

A

-R2 Ric); X R1D wherein X is O and RIB-RID and R3A-R3E are independently selected from hydrogen, hydroxy, methoxy, and amino or any 2 adjacent RI and/or R 3 groups from RIB-RID and R3A-R3E form a dioxolanyl group;

R

2 is an optionally substituted aryl or optionally substituted heteroaryl group; P %Opcrmjl2lMl :26416 2 Illooc-2 1/11 /21)6 S-12- 0 CD A' is C=O and A is a single bond.

In a further aspect, the invention provides a method of preparing a compound of formula 00 R1A R3 RIC X R 2

R

1D RRo wherein X is O, NH or NR, (wherein R is H, sulfonyl, Ci.6alkyl, C2-6alkenyl, C 2 .6alkynyl, or Ci- 7acyl, or an aryl or heteroaryl group); A' is independently selected from a single bond or C=O; RIA-RID are independently selected from hydrogen, hydroxy, optionally substituted alkoxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted acyloxy, amino, optionally substituted alkylamino, optionally substituted dialkylamino, optionally substituted acylamino, or any 2 adjacent RIA-RID together form -O-CH 2

R

2 and R 3 are independently optionally substituted aryl or optionally substituted heteroaryl groups; said method comprising the steps of: a) coupling a compound of formula with an alkyne of formula in the presence of a nickel or palladium coupling agent P 1Op rV"a12fl(N l 22h 160 .111 I dOC.2 I/I 1112(96 -13- 00 R B Hal k Rlc XM1 RlD (2) Hal is I ,Br or CI; M, is a metal or a metal species thereof, said metal selected from the group consisting of Li, Na, K, Mg, Cs and Ba;

M

2 is a metal, or a metal species thereof, said metal selected from the group consisting of Mg, Zn, Cu, B, Si, Mn, Sn, Ge and Al; X is O or NR (wherein R is sulfonyl, C1.6alkyl, C2.

6 alkenyl, C2.6alkynyl, or C 1 7 acyl); b) reacting in situ the resulting coupled product with R 3 wherein R 3 is an optionally substituted aryl group or optionally substituted heteroaryl group and wherein L is a leaving group, optionally in the presence of carbon monoxide.

In yet another aspect, the invention provides a method for preparing a compound of formula R1A R3 R1B

A'

Rlc X R 2 R1D wherein X is S; P opeVtalX2tMl22f4161) 2)II doc 21/11/20li6 S-14- U A' is selected from a single bond or C=O; RIA-RID are independently selected from hydrogen, hydroxy, optionally substituted alkoxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted acyloxy, amino, optionally substituted alkylamino, N optionally substituted dialkylamino, optionally substituted acylamino, or any two adjacent RIA-RID together form -O-CH 2

R

2 and R 3 are independently optionally substituted aryl or optionally substituted heteroaryl groups.; said method comprising the steps of: a) coupling a compound of formula with a compound of formula in the presence of a nickel or palladium coupling agent R1A

R

1 B Hal

R

RiC SPs R1D (4) wherein RIA-RID, Hal., M 2 and R 2 are as above, and Ps is a sulfur protecting group capable of stabilizing a positive charge; b) cyclising the resulting coupled product in the presence of a Hal' producing reagent to give Pop'rm IalUt 122MI 6 2U II doc-2 I llI21)6 R1A S/Hal R1B 2 IN Ric S 00 R1

D

C wherein 0Hal is Cl, Br or I; c) coupling with either the moiety R 3 or R 3 wherein R 3 is an optionally substituted aryl or optionally substituted heteroaryl group.

In another aspect, the invention provides a method for preparing a compound of formula R1A R3 R1B A-R2

()I

R1D wherein A is selected from a single bond or C=0; RIA-RID are independently selected from hydrogen, hydroxy, optionally substituted alkoxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted acyloxy, amino, optionally substituted alkylamino, optionally substituted dialkylamino or any 2 adjacent RIA-RID together form

-O-CH

2 is an optional double bond;

R

2 and R 3 are independently optionally substituted aryl or optionally substituted heteroaryl groups; PWcrIml2(X I M22h160l( 201 doc2 I/11/2Ml -16said method comprising the steps of: reacting compound with compound or reacting compound 6(a) with compound 7(a).

RID 0 M R 2 (7) R1A (6a) (7a) to form a compound (9) P op r Jll2(XMQ612M Ih1 2()1 I Joc-2 1/21 I/I -17-

O

o R1A 00 RIAR SR2 Ro O Cl RID 0 (9) wherein M is Li, Na, K or MgHal (Hal is Br, Cl or I); b) treating compound with a metal hydride in the presence of a palladium coupling agent; c) coupling the resulting product with R 3 -Hal or R 3 -C(0)-Hal (wherein Hal is Cl, Br or I) to provide either compound (10) or and R1A 1A R 1B e R

R

3 R3 Ric

R

2 RiC RID 0 RID 0 (11) cyclising (10) or (11) under acidic conditions to form an indanone and optionally treating the cyclised product with an oxidising agent to form an indenenone.

P Opcrntal\2(XOi 22641h I2 21 dc-2 1/21 i/2(6M 18- In yet a further aspect, the invention provides a method for preparing a compound of Formula R1A R3 A--R2 x R1C R1D wherein; X is O, S or NR (wherein R=H, C-.6alkyl, C2-6alkenyl, C2-6alkynyl, or C(O)C.-6alkyl); RIA-RID are as previously defined; A is C=O;

R

2 and R 3 are independently optionally substituted aryl or optionally substituted heteroaryl groups; comprising the steps of a) coupling a compound (12) with compound (13) Hal (12) wherein Hal is Cl, Br, or I; to form a compound of formula (14); (13) P lopeMVnlal))6I\22M 160 2011 doc-21/lI /2X06 -19- (14) b) when X is S, protecting the thiol with a sulfur-protecting group c) reacting (14) with a compound M1---R3 wherein MI is Li, Na, K, Mg, Cs or Ba, and R 3 is an optionally substituted aryl or optionally substituted heteroaryl group; to form R1A HO R3

RIB

I R 2 Ric T

XP

Rio wherein when X is O or NH, then P is H and when X is S, P is a sulfur protecting group and when XP is NR, R is a hydrogen, sulfonyl, Ci-6alkyl, C 2 6 alkenyl, C2- 6 alkynyl, C 1 -7acyl or an aryl or heteroaryl group; d) treating (15) with a Hal producing reagent, to afford cyclisation.

Still another aspect of the invention provides a compound of Formula II P %oPcfmMJA 2:k).26n IN) NI I dm-2111 1i12M o

IN

0

(II)

R1A substituted dialkylamino or 2 adjacent RAR2RD are R1D wherein RIA-RID are independently hydrogen, hydroxy, optionally substituted alkoxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted acyloxy, amino, optionally substituted alkylamino, optionally substituted dialkylamino or 2 adjacent RIA-RID are

O-CH

2

R

2 is an optionally substituted aryl or optionally substituted heteroaryl group; A is C=O.

Still yet a further aspect of the invention relates to compounds of Formula (III) R3

R

5

A

(III)

R6 S

RZ

wherein

R

2 and R 3 are independently optionally substituted aryl or optionally substituted heteroaryl groups; A' is CO;

R

5 and R 6 can independently be hydrogen, optionally substituted alkyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl or P loM~AIUM)l 22641 W21111 dx.21/11/2l6 0 -21-

O

optionally substituted alkenyl; is an optional double bond.

Other aspects of the invention relate to combinatorial libraries of compounds comprising oO at least two compounds of Formula (II) or (III).

Ci Still other aspects of the invention relate to the use of the compounds of the present Sinvention in the manufacture of medicaments, and methods thereof, in the treatment of Ci conditions requiring tublin polymerization inhibitors.

BRIEF DESCRIPTION OF THE DRAWINGS The drawings which follow form part of the present specification and are included to further demonstrate certain aspects of the present invention. The invention may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

Figure 1 schematically depicts a generalised "one pot" synthesis of benzofused furans and indoles.

Figure 2 schematically depicts a generalised "one pot" synthesis of benzofused furans and indoles.

Figure 3 schematically depicts a generalised synthesis of benzofused thiophenes.

Figure 4 schematically depicts a generalised synthesis of benzofused thiophenes, including compound from US Patent No. 5,886,025.

Figure 5 schematically depicts a generalised synthesis of benzofused thiophenes.

Figure 6 schematically depicts a generalised synthesis of benzofused thiophenes.

Figure 7 schematically depicts a generalised "one pot" synthesis of aryl substituted P ~p ~I2E~ IN) 6 20:1 eCx 21111/(X.6 -22cc, P-alkenyl carbonyl compounds, for the preparation of indanones and indenones.

Figure 8 schematically depicts a generalised synthesis of benzofused furans, indoles 00 and thiophenes.

Fiture 9 schematically depicts a generalised synthesis of benzofused thiophenes.

Figure 10 schematically depicts a generalised synthesis of thiophenes.

Figure 11 depicts the structures of some of the preferred compounds of the present invention which posses TPI activity.

DETAILED DESCRIPTION OF THE INVENTION As used herein, the term "alkyl", used either alone or in compound words such as "alkylamino' and "dialkylamino" etc, denotes straight chain, branched or cyclic alkyl, preferably C 1 20 alkyl, eg C 1 0 or CI- 6 alkyl. Examples of straight chain and branched alkyl include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, amyl, isoamyl, sec -amyl, 1 ,2 -dim ethyl propyl, 1,1 -dimethyl-propyl, hexyl, 4-methylpentyl, 1Imethylpenty,, 2-methylpentyl, 3-methylpentyl, 1,1-dimethylbutyl, 2 ,2-dimethylbutyl, 3,3 dimethylbutyl, 1 ,2-dimethylbutyl, 1 ,3-dimethylbutyl, 1,2,2 ,-trimethylpropyl, 1,1 ,2trimethyiprop yl, heptyl, 5-methylhexyl, I -methylhexyl, 2,2-dimethylpentyl, 3,3dimethylpen-tyl, 4,4-dimethylpentyl, 1 ,2-di methyl pentyl, 1 ,3-dimethylpentyl, 1,4dimethyl-pentyl, 1, 2,3 -trim ethylbutyl, 1,1 ,2-trimethylbutyl, 1,1 ,3-trimethylbutyl, octyl, 6methylheptyl, 1-methyiheptyl, 1,1,3,3-tetramethylbutyl, nonyl, 6- or 7methyl-octyl, 4- or 5-ethylheptyl, 2- or 3-propylhexyl, decyl, 7- and 8-methylnonyl, 5- or 6-ethyloctyl, 3- or 4-propylheptyl, undecyl, 8- or 9-methyldecyl, 6- or 7-ethylnonyl, 4- or 5-propylocytl, 2- or 3-butylheptyl, I -pentylhexyl, dodecyl, 4-, 9- or 10-methylundecyl, 7- or 8-ethyldecyl, 4-, or 6-propylnonyl, 3- or 4-butyloctyl, 1-2-pentyiheptyl and the like. Examples of cyclic alkyl include mono- or polycyclic alkyl groups such as cyclopropyl, cyclobutyl, P koper Vnll20tX)6\ 2264 IM)201 oc.-21/112 )6 0 -23- U cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl and the like.

Where an alkyl group is referred to generally as "propyl", butyl" etc, it will be understood

O

NC that this can refer to any of straight, branched and cyclic isomers where appropriate.

"Alkoxy" refers to an alkyl when conveniently bonded to an oxygen atom. An alkyl oO group may be optionally substituted by one or more optional substituents as herein defined.

Accordingly, "alkyl" as used herein is taken to refer to optionally substituted alkyl.

(N

C Optional substituents for an alkyl group include hydroxy, halo, alkoxy, phenyl, benzyl, Sphenoxy, benzyloxy, carbonyl, amino, acyl, acyloxy, alkylamino, dialkylamino, C1 acylamino. Particularly preferred optional substituents include those wherein the alkyl moiety of the substituent is C 1 6 alkyl.

Optionally substituted alkoxy, alkylamino, dialkylamino, refers to the optional substitution of the "alkyl" moiety. Similarly, optionally substituted, acyl, acylamino and acyloxy refer to the optional substitution of the alkyl or aryl moieties of the acyl group.

The term "alkenyl" as used herein denotes groups formed from straight chain, branched or cyclic hydrocarbon residues containing at least one carbon to carbon double bond including ethylenically mono-, di- or poly-unsaturated alkyl or cycloalkyl groups as previously defined, preferably Cl- 20 alkenyl (eg Cl-.o or Ci-6). Examples of alkenyl include vinyl, allyl, 1-methylvinyl, butenyl, iso-butenyl, 3-methyl-2-butenyl, 1-pentenyl, cyclopentenyl, 1-methyl-cyclopentenyl, 1-hexenyl, 3-hexenyl, cyclohexenyl, 1-heptenyl, 3-heptenyl, 1-octenyl, cyclooctenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 1-decenyl, 3decenyl, 1,3-butadienyl, 1,4-pentadienyl, 1,3-cyclopentadienyl, 1,3-hexadienyl, 1,4hexadienyl, 1,3-cyclohexadienyl, 1,4-cyclohexadienyl, 1,3-cycloheptadienyl, 1,3,5cycloheptatrienyl and 1,3,5,7-cyclooctatetraenyl. An alkenyl group may be optionally substituted by one or more optional substituents as herein defined.

The term "alkynyl" as used herein denotes groups formed from straight chain or branched hydrocarbon residues containing at least one carbon to carbon triple bond.

Particularly preferred alkynyl groups are C 2

-C

6 alkynyl. An alkynyl group may be optionally substituted by one or more optional substituents as herein defined.

P p)pcnAI\2(Ll\122MIW 62)11 doC-2 1/2/I)6 0 -24-

O

U Unless indicated otherwise, the term "halogen", "halo" "halide" etc. denotes fluorine, chlorine, bromine or iodine (fluoro, chloro, bromo or iodo) (fluoride, chloride, 0 bromide iodide).

00 The term "aryl" includes single, polynuclear, conjugated and fused residues of aromatic hydrocarbon ring systems. Examples of hydrocarbon based "aryl" include C phenyl, biphenyl, terphenyl, quaterphenyl, naphthyl, tetrahydronaphthyl, anthracenyl, O dihydroanthracenyl, benzanthracenyl, dibenzanthracenyl, phenanthrenyl, fluorenyl, C pyrenyl, idenyl, azulenyl, chrysenyl.

The term "heteroaryl" includes cyclic (single, polynuclear, fused or conjugated) hydrocarbon residues where one or more carbon atoms are replaced by a heteroatom and form an aromatic residue where two or more carbon atoms are replaced, this may be by the same heteroatom or different heteroatoms. Suitable heteroatoms include O, N, S and Se.

Suitable heteroaryl include furanyl, thieuyl, pyrrolyl, indolyl, pyridyl, pyridazinyl, pyrazoyl, pyrazinyl, thiazolyl, pyrimidinyl, quinolinyl, isoquinolinyl, benzofuranyl and benzothienyl.

An aryl or heteroaryl group may be optionally substituted by one or more substituents. Preferred optional substituents include hydrogen, hydroxy, alkoxy, alkyl, alkenyl, alkynyl, acyloxy, amino, alkylamino, dialkylamino or any 2 adjacent positions are substituted to together form -O-CH 2 An aryl or heteroaryl group may also be optionally fused to a cyclic or polycyclic group (saturated or unsaturated), which itself may be further optionally substituted as described for "alkyl" above.

The term "acyl" either alone or in compound words such as "acyloxy", or "acylamino" etc, denotes a group containing the moiety C=0 (and not being a carboxylic acid, ester or amide or thioester) Preferred acyl includes wherein R is hydrogen or an alkyl, or aryl preferably a C 1 20 residue. Ci.

7 acyl refers to an acyl group that counts the carbonyl group as one carbon atom. Examples of acyl include formyl; straight chain or branched alkanoyl such as, acetyl, propanoyl, butanoyl, 2-methylpropanoyl, pentanoyl, 2,2-dimethylpropanoyl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl, undecanoyl, P \opcIr\nll 0 M1264 160 2ZOi l doc21 I/l 1/21)6 )ddodecanoyl, tridecanoyl, tetradecanoyl, pentadecanoyl, hexadecanoyl, heptadecanoyl, Soctadecanoyl, nonadecanoyl and icosanoyl; cycloalkylcarbonyl such as

O

C cyclopropylcarbonyl cyclobutylcarbonyl, cyclopentylcarbonyl and cyclohexylcarbonyl; aroyl such as benzoyl, toluoyl and naphthoyl; aralkanoyl such as phenylalkanoyl (e.g.

00 phenylacetyl, phenylpropanoyl, phenylbutanoyl, phenylisobutylyl, phenylpentanoyl and phenylhexanoyl) and naphthylalkanoyl naphthylacetyl, naphthylpropanoyl and C, naphthylbutanoyl]. The term "acyloxy" refers to an acyl group covalently bonded to an Soxygen atom.

As used herein, the term "protecting group", refers to an introduced functionality which temporarily renders a particular functional group inactive under certain desired conditions. Suitable protecting groups are known to those skilled in the art, for example as described in Protective Groups in Organic Synthesis Greene and P.G.M. Wutz, Wiley Interscience, New York, 3 r d edition).

As used herein, the term "leaving group" refers to a chemical group which is displaced by a nucleophile. Suitable leaving groups include those with the ability to stabilise the negative charge which it carries such as the halogens I, Br, Cl), triflate trifluoromethane sulfonyl), acetate and sulfonates (eg. tosylate, mesylate, nosylate etc). Some preferred leaving groups are I, Br, Cl and trifluoromethane sulfonyl.

In preferred embodiments of the invention, RIA-RID include hydrogen, hydroxy, optionally substituted C 1 .6alkoxy, optionally substituted Ci.

6 alkyl, optionally substituted

C

2 6 alkenyl, optionally substituted C 2 6 alkynyl, optionally substituted C(O)-Ci.

6 alkyl, amino, optionally substituted Ci.

6 alkylamino, optionally substituted diCi.

6 alkylamino or 2 adjacent RIA-RID form a dioxolanyl group (-O-CH 2 Particularly preferred RIA-RID include: hydrogen, hydroxy, methoxy, ethoxy, propoxy, butoxy, methyl, ethyl, propyl, butyl, acetyl, acetyloxy, amino, methylamino, ethylamino, propylamino butylamino, dimethylamino, diethylamino, dipropylamino, dibutylamino or 2 adjacent RIA-RID form a dioxolanyl group.

In yet other preferred embodiments of the invention, R 2 and R 3 can be independently an optionally substituted phenyl group of formula P cperimI\2IX)6 122616M 0 2n1I I dc- l/11 I/2i6

NO

-26- R2C c-R2 D R 28 S(i) 00oo R2E R2A

(-I

wherein R2A-R2E are independently selected from hydrogen, hydroxy, optionally substituted alkoxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted acyloxy, amino, optionally substituted alkylamino, optionally substituted dialkylamino, optionally substituted acylamino or where any two adjacent R 2 A-R2D together form -O-CH 2 Preferred R2A-R2E are as for RIA- RID described above. Preferred forms of formula are where R2A-R2E are independently hydrogen, hydroxy, methoxy, amino or any two adjacent R2A-R2D together form -O-CH 2 In the process for the preparation of compounds of Formula compounds and are derived from their respective phenol or protected amine and terminal alkyne respectively. The starting phenol or aniline and terminal alkyne can be coupled together under conditions which allow for the heteroannulation to spontaneously occur so as to form the target benzo[b]furan or indole in a "one-pot" synthetic strategy. Thus, the metal based compound required to form must be such that the phenol or protected amine is deprotonated to form the group -OM 1 or NHMI.

Suitable MI are based on Li, Na, K, Mg, Cs and Ba as well as species formed therefrom, for example from Grignard reagents Ci.

4 alkyl MgHal (Hal I, Cl or Br) Suitable metal species include MgCl, MgBr or MgI. Formation of can be effected by treating the corresponding phenol or protected amine with, for example, Li 2

CO

3 Na 2

CO

3

K

2

CO

3 MgCC 3 Cs 2

CO

3 BaCO 3 MeMgCI, EtMgCI, MeMgBr, EtMgBr, MeMgI and EtMgI.

P opcnmnlUIIll'\ 22 1 12011 i doc 2l)/ /2I I/ 0 -27-

O

M

2 can be a hydrogen atom or metal species used in any palladium or nickel cross-

O

CN coupling protocols known in the art, (for example, Stille, Suzuki or Negishi cross-coupling reactions using stannanes (eg aryl or alkylstannanes, boronic acids/esters or zinc based OO compounds eg. ZnCI) for example based on Mg, Zn, Cu, B, Si, Mn, Sn, Ge or Al.

E Particularly suitable M 2 include ZnC1, (alkyl) 3 Sn, (aryl)3Sn, B(OR) 2 (R is, eg, H alkyl, CN alkenyl or alkynyl), MgBr, MgCl and MgI.

C In a particularly preferred form of this aspect of the invention both MI and M 2 are derived from a Grignard reagent such as an alkyl magnesium halide eg. Ci-4alkylMgBr, (Cl) or Suitable M, and M 2 thus include MgCI, MgBr and MgI.

Where X is NR in formula the nitrogen atom of the starting aniline is suitably protected by a nitrogen protecting group. Suitable nitrogen protecting groups are known to those skilled in the art of organic synthesis and include acyl groups (eg acetyl, trifluoroacetyl), phenyl, benzyl and benzoyl. Other suitable nitrogen protecting groups may be found in Protective Groups in Organic Synthesis, T. W. Greene and P. Wutz, John Wiley Son, ,3 rd Edition.

The coupling agent used in step of the process for the preparation is preferably a nickel or palladium based coupling agent. Suitable coupling agents are known in the art and include Pd(PPh 3 2 C1 2 Pd(PPh 3 4 Pd(dibenzylideneacetone) 3 and PdCI 2

(CH

3

CN)

2 The leaving group of R 3 -L can be any suitable leaving group known to the skilled person. In a preferred embodiment, where R 3 is an optionally substituted aryl or optionally substituted heteroaryl, L can be a halogen such as iodine, chlorine or bromine, a triflate, or a sulfonate (eg tosylate, mesylate, brosylate nosylate etc).

Two preferred embodiments of the process for preparing compounds of formula are schematically depicted in Figures 1 and 2.

The preparation of benzo[b]thiophenes of formula is effected using a variation P WopoemalUfIM\&I 224 111211 doe2111 lrIm6

IO

-28- 0 U of the methods described for the benzo[b]furans and indoles of formula above. In particular, the sulfur atom, X, must be protected by a suitable protecting group to C circumvent competitive coupling of a thiolate to the aryl halide to afford xanthones.

Suitable sulfur protecting groups are those which are capable of stabilizing a positive 00 charge. Examples include benzyl, allyl, acetyls and thioacetals.

SAs used herein a Half producing agent is an agent which can effectively act as a O Hal" source. Examples of Hal+ producing agents include 12, Br 2 Cl 2 IBr, IC1, CI chloroacetamide, iodoacetamide, N-chlorosuccinamide, N-bromosuccinamide and Niodosuccinamide.

Suitable M 2 for compound and coupling agents for the preparation of compounds of Formula may include those described above.

The coupling of with the moiety R 3 or R 3 to produce can be carried out via palladium-mediated coupling and/or metallation techniques as known in the art.

For example, lithiation of (eg using nBuLi) allows for coupling with R 3 -C(O)-Hal (Hal is I, Br or Cl, preferably Cl). In another embodiment, Negishi coupling of with R 3 ZnCI (derived from reaction of R 3 -Li with ZnC1) gives access to compounds of formula where A is a single bond. In another embodiment, palladium-mediated couplings such as Suzuki or Stille couplings can be used to access compounds of formula Some preferred embodiments for the preparation of Compounds of Formula are shown in Figures 3, 4, 5 and 6.

In the preparation of compounds of Formula step involves the use of a metal hydride. Suitable metal hydrides are those which react with the triple bond to form an intermediate metallated vinyl group without reducing the adjacent carbonyl group and may include trialkylstannanes (eg trimethyl- or tributyl tin hydride), aryl stannanes (eg triphenyl tin hydride), copper hydride, diisobutylaluminiumhydride, or borohydrides (eg catechol borane).

P oper, &11 W161 2264160 2011 da ,.211 11214 -29-

O

U The cyclization step employs acidic conditions, which can be either a H source Sor a Lewis acid. Suitable acids include HCI, H 2

SO

4

BF

3

AICI

3 methanesulphonic acid Cetc. A preferred oxidizing agent is DDQ.

O A schematic representation of one method for the preparation of compounds of 0.

Formula is depicted in Figure 7.

0 In the preparation of compounds of Formula the coupling of (12) and (13) can C1 be carried out using suitable metallation techniques known in the art. Thus the coupling can be carried out in the presence of n-BuLi sec-BuLi, t-BuLi or alkylMghalides such as iPrMgHalide A schematic representation for a method of preparing compounds of Formula is shown in Figure 8.

Compounds of Formula (II) can be prepared by coupling of a suitably protected thiol aldehyde under Negishi conditions, lithiation and coupling with the appropriate ZnCI acetylide. Cyclisation using a Hal producing agent, eg iodocyclization using 12, affords access to Formula (II).

A schematic representation for a method of preparing compounds of Formula (II) is shown in Figure 9.

Compounds of Formula (III) can be prepared by palladium-mediated coupling of an appropriately protected butynyl sulfide with R 2 for example under Sonagashira conditions using CuI. Other palladium-mediated coupling procedures such as Stille, Suzuki, and Negishi conditions can also be used. Cyclization can be effected using a Hal+ producing agent as described herein. Coupling of the resulting Hal-dihydrothiophene with an R 3 moiety, for example, using ZnCl-R3. provides products where A' is a single bond.

Alternatively, coupling with R 3 -C(O)-Hal in the presence of nBuLi as described herein provides access to compounds of Formula (III) wherein A' is C=0.

01-04-'08 16:21 FROM- T-512 P009/014 F-140 00 0 0 ci 0 koftftNBALatIMlG 264I5E V d53spPo fin) A schematic representation for a method of preparing compounds of Formula IllI is depicted in Figure An important aspect of the present invention relates to compounds which may possess tubulin binding activity. Compounds which possess tubulin binding activity may act as anti-mitotic agents and may be effective in targeting tumour vasculature.

Thus, the invention also provides compounds of Formula (1,11) wherein X is 0; and is an optional double bond; RIB-RID and R3A-R3E arc independently selected from hydrogen, hydroxy, methoxy, and amino or any 2 adjacent RIB-RID and R3A-R3E form a dioxolanyl group;

R

2 is an optionally substituted aryl or optionally substituted heteroaryl group, preferably an optionally substituted phenyl group of formula as herein described.

A' is C=0; and A is a single bond.

COMS ID No: ARCS-184998 Received by IP Australia: Time 16:22 Date 2008-04-01 01-04-'08 16:21 FROM- T-512 P010/014 F-140 00 0 0 0 \OpPTHTMAlU2ll\Z2I MW2 (16 2d-21A6rl'A -31- In a preferred form is a double bond.

In another preferred embodiment of when R 2 is a phenyl group of formula at least one of RIB-RID and R 3 A-R3Fis a hydroxy or amino group which can be derivatized to form a salt or prodrug, such as an ester or amide, preferably a disodium phosphate ester.

In another preferred embodiment of when Rz is a phenyl group of formula the phenyl group bears vicinal hydroxy and methoxy groups.

Yet other preferred compounds which may possess tubulin binding activity have the formula (IV): R3c Ra R (IV R4

A

R2 R20 (IV) X is O, S or NR" (wherein R" is aryl, heteroaryl, aroyl, heteroaroyl, acyl, benzyl, alkyl, alkenyl, alkynyl or sulfonyl) A is a single bond, C=0 or O, S or NR (wherein R is hydrogen, C 1 -6 hydrogen, Cl.

6 alkyl,

C

26 alkenyl, C2_alkynyl or C Lacyl).

R2A-R2E and R3A-R3E are as hereinbefore described.

COMS ID No: ARCS-184998 Received by IP Australia: Time 16:22 Date 2008-04-01 P op~nmlUI.ll012264160 2111 ldo2 IlI ;I21LU 0 -32- <U R2A-R2E and R3A-R3E are as hereinbefore described.

CI R 4

-R

7 are independently selected from hydrogen, hydroxy, alkoxy, alkyl, alkenyl, alkynyl, amino, alkylamnino, dialkylamino, acyl, acylamino, heteroaryl and aryl.

OO

00 The compounds of Formula (IV) can be prepared using iodocyclisation procedures known N1 for forming compounds where X 0 or N (Bev, S.P. et al. Chem. Commun. 1996, 1007 O and Knight, D.W. et al. Chem. Commun. 1998, 2207) or for where X S, analogous (N methodology as described herein.

Couplings of the aryl groups R 2 and R 3 can be performed as described herein and for where A is O, S or NH, see Bavanno, et al, Curr. Org. Chem. 1997, 3, 287 and references cited therein.

Some examples of compounds of the invention which possess tubulin-binding activity are depicted in Figure 11.

Certain compounds of the invention having tubulin binding activity or which have anti-tumour vasculature activity, may be useful in methods of therapy. In particular these compounds may be used for treating tumours. As used herein the term "tumour" is used to define any malignant cancerous growth, and may include leukemias, melanomas, colon, lung, ovarian, skin, breast, prostate, CNS, and renal cancers, as well as other cancers.

The compound of the invention having tublin binding activity may also be used in the treatment of solid tumours, eg. breast cancer.

The invention also provides for the use of a compound of formula or (II)-(IV) in the manufacture of a medicament for treating tumours.

There is also provided a method of treatment of solid tumours comprising the administration of an effective amount of a compound of formula or to a subject in need thereof.

P opcrVnaIl X)61122641IN) I d l, 21/I 1/2~n6 -33- The compounds of the invention may be particularly useful in combination therapy, eg. combining the treatment with other chemotherapeutic or radiation treatments.

INO

00 However, it will be understood that the compounds of the invention can be used in r the treatment of any disease for which tublin polymerization plays a crucial role.

I

I

I

SCompounds of the invention which possess bioactivity, such as tubulin binding activity, can be formulated as a composition, particularly a pharmaceutical composition, together with a pharmaceutically acceptable additive.

The compounds of the invention are administered to the subject in a treatment effective amount. As used herein, a treatment effective amount is intended to include at least partially attaining the desired effect, or delaying the onset of, or inhibiting the progression of, or halting or reversing altogether the onset or progression of the particular disease of condition being treated.

As used herein, the term "effective amount" relates to an amount of compound which, when administered according to a desired dosing regimen, provides the desired therapeutic activity. Dosing may occur at intervals of minutes, hours, days, weeks, months or years or continuously over any one of these periods. Suitable dosages lie within the range of about 0.1 ng per kg of body weight to 1 g per kg of body weight per dosage. The dosage is preferably in the range of 1 pig to 1 g per kg of body weight per dosage, such as is in the range of 1 mg to 1 g per kg of body weight per dosage. In one embodiment, the dosage is in the range of 1 mg to 500 mg per kg of body weight per dosage. In another embodiment, the dosage is in the range of 1 mg to 250 mg per kg of body weight per dosage. In yet another preferred embodiment, the dosage is in the range of 1 mg to 100 mg per kg of body weight per dosage, such as up to 50 mg per body weight per dosage.

Suitable dosage amounts and dosing regimens can be determined by the attending physician and may depend on the particular condition being treated, the severity of the condition as well as the general age, health and weight of the subject.

P %OMM J%21122 16A )111 0oc2 ]1 I/2(XM -34- The active ingredient may be administered in a single dose or a series of doses.

N, While it is po:ssible for the active ingredient to be administered alone, it is preferable to present it as a composition, preferably as a pharmaceutical composition. The formulation 00 of such compositions is well known to those skilled in the art. The composition may rcontain any suitable carriers, diluents or excipients. These include all conventional c, solvents, dispersion media, fillers, solid carriers, coatings, antifungal and antibacterial Sagents, dermal penetration agents, surfactants, isotonic and absorption agents and the like.

C, It will be understood that the compositions of the invention may also include other supplementary physiologically active agents.

The carrier must be pharmaceutically "acceptable" in the sense of being compatible with the other ingredients of the composition and not injurious to the subject.

Compositions include those suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parental (including subcutaneous, intramuscular, intravenous and intradermal) administration. The compositions may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy.

Such methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more accessory ingredients. In general, the compositions are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then if necessary shaping the product.

Compositions of the present invention suitable for oral administration may be presented as discrete units such as capsules, sachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be presented as a bolus, electuary or paste.

A tablet may be made by compression or moulding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable P opcr\Val\2i*\I 2h4 164fi 2Il1 doc.2 li 11/21 machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder (e.g inert diluent, preservative disintegrant sodium cN starch glycolate, cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose) surface-active or dispersing agent. Moulded tablets may be made by moulding 00 in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to C-1 provide slow or controlled release of the active ingredient therein using, for example, O hydroxypropylmethyl cellulose in varying proportions to provide the desired release rC profile. Tablets may optionally be provided with an enteric coating, to provide release in parts of the gut other than the stomach.

Compositions suitable for topical administration in the mouth include lozenges comprising the active ingredient in a flavoured base, usually sucrose and acacia or tragacanth gum; pastilles comprising the active ingredient in an inert basis such as gelatin and glycerin, o: sucrose and acacia gum; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

Compositions suitable for topical administration to the skin may comprise the compounds dissolved or suspended in any suitable carrier or base and may be in the form of lotions, gel, creams, pastes, ointments and the like. Suitable carriers include mineral oil, propylene glycol, polyoxyethylene, polyoxypropylene, emulsifying wax, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. Transdermal patches may also be used to administer the compounds of the invention.

Compositions for rectal administration may be presented as a suppository with a suitable base comprising, for example, cocoa butter, glycerin, gelatin or polyethylene glycol.

Compositions suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active ingredient such carriers as are known in the art to be appropriate.

P \opeuV\mnl2()6 122646 I12011) d -2 1/1 1/216 -36-

O

SCompositions suitable for parenteral administration include aqueous and non- CK aqueous isotonic sterile injection solutions which may contain anti-oxidants, buffers, bactericides and solutes which render the composition isotonic with the blood of the 00 intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The compositions may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a Sfreeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.

Preferred unit dosage compositions are those containing a daily dose or unit, daily sub-dose, as herein above described, or an appropriate fraction thereof, of the active ingredient.

It should be understood that in addition to the active ingredients particularly mentioned above, the compositions of this invention may include other agents conventional in the art having regard to the type of composition in question, for example, those suitable for oral administration may include such further agents as binders, sweeteners, thickeners, flavouring agents disintegrating agents, coating agents, preservatives, lubricants and/or time delay agents. Suitable sweeteners include sucrose, lactose, glucose, aspartame or saccharine. Suitable disintegrating agents include corn starch, methylcellulose, polyvinylpyrrolidone, xanthan gum, bentonite, alginic acid or agar.

Suitable flavouring agents include peppermint oil, oil of wintergreen, cherry, orange or raspberry flavouring. Suitable coating agents include polymers or copolymers of acrylic acid and/or methacrylic acid and/or their esters, waxes, fatty alcohols, zein, shellac or gluten. Suitable preservatives include sodium benzoate, vitamin E, alpha-tocopherol, ascorbic acid, methyl paraben, propyl paraben or sodium bisulphite. Suitable lubricants include magnesium stearate, stearic acid, sodium oleate, sodium chloride or talc. Suitable time delay agents include glyceryl monostearate or glyceryl distearate.

P %OprWIQM6 12264160 2011 dO2 11 !21XI

\D

0 -37-

O

SThe novel bioactive compounds of the invention can be administered to a subject as a salt or prodrug thereof. The term "salt, or prodrug" includes any pharmaceutically 0 CI acceptable salt, ester, solvate, hydrate or any other compound which, upon administration to the recipient is capable of providing (directly or indirectly) a compound as described Io 00 herein. However, it will be appreciated that non-pharmaceutically acceptable salts also fall r.within the scope of the invention since these may be useful in the preparation of C1 pharmaceutically acceptable salts. Any compound that is a prodrug of a compound of 0 formula is within the scope and spirit of the invention. The term "pro-drug" is used in i its broadest sense and encompasses those derivatives that are converted in vivo to the compounds of the invention. Such derivatives would readily occur to those skilled in the art, and include, for example, compounds where a free hydroxy group is converted into an ester, such as an acetate or phosphate ester, or where a free amino group is converted into an amide. Procedures for esterifying, eg. acylating, the compounds of the invention are well known in :he art and may include treatment of the compound with an appropriate carboxylic acid, anhydride or chloride in the presence of a suitable catalyst or base. A particularly preferred prodrug is a disodium phosphate ester. The disodium phosphate ester of novel compounds of the invention may be useful in targeting tumour vasculature and thus may provide a means of selective delivery of the compounds to the body. The disodium phosphate ester may be prepared in accordance with the methodology described in Pettit, G. et al, Anticancer Drug Des., 1995, 10, 299.

Suitable pharmaceutically acceptable salts include, but are not limited to salts of pharmaceutically acceptable inorganic acids such as hydrochloric, sulphuric, phosphoric nitric, carbonic, boric, sulfamic, and hydrobromic acids, or salts of pharmaceutically acceptable organic acids such as acetic, propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric, maleic, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulphonic, toluenesulphonic, benezenesulphonic, salicyclic sulphanilic, aspartic, glutamic, edetic, stearic, palmitic, oleic, lauric, pantothenic, tannic, ascorbic and valeric acids.

Base salts include, but are not limited to, those formed with pharmaceutically acceptable cations, such as sodium, potassium, lithium, calcium, magnesium, ammonium P NW CTI a2PWA% 12264 lW 211 do:- 2 1 1,20, o -38- 0 Dand alkylammonium. In particular, the present invention includes within its scope cationic salts eg sodium or potassium salts, or alkyl esters (eg methyl, ethyl) of the phosphate N, group.

ID

00 Basic nitrogen-containing groups may be quartemised with such agents as lower alkyl halide, such as methyl, ethyl, propyl, and butyl chlorides, bromides and iodides; C dialkyl sulfates like dimethyl and diethyl sulfate; and others.

NI The compounds of the invention may be in crystalline form either as the free compounds or as solvates hydrates) and it is intended that both forms are within the scope of the present invention. Methods of solvation are generally known within the art.

It will also be recognised that compounds of the invention may possess asymmetric centres and are therefore capable of existing in more than one stereoisomeric form. The invention thus also relates to compounds in substantially pure isomeric form at one or more asymmetric centres eg., greater than about 90% ee, such as about 95% or 97% ee or greater than 99% ee, as well as mixtures, including racemic mixtures, thereof. Such isomers may be prepared by asymmetric synthesis, for example using chiral intermediates, or mixtures may be resolved by conventional methods, eg., chromatography, or use of a resolving agent.

The synthetic methods and processes described herein are amenable to combinatorial chemistry to produce libraries of compounds for biological screening.

Traditionally, drug candidates have been synthesized individually, this being a time consuming and laborious process if the synthetic sequence contains even just a few steps and large numbers of compounds are to be evaluated for their biological activity.

Combinatorial synthesis is an emerging technique for effecting the generation of large libraries of molecules and has been successfully exploited in the synthesis and evaluation of small organic libraries. These libraries and their starting substrates may exist as molecules in free solution or preferably, linked to a solid phase, for example, beads, pins, microtitre plates (wells) or microchips which can be polymeric, glass, silica or other P \pfr .l2(Xl 61264 I 1 I0 1 d -2 /I I 2xt6 -39-

U

a suitable substrate. Chemical diversity can be achieved by either parallel or split (split and mix) syntheses wherein each step has the potential to afford a multitude of compounds.

CK, Solution phase libraries may be prepared via parallel syntheses wherein different compounds are synthesised in separate reaction vessels in parallel, often in an automated

INO

00 fashion. Alternatively, attachment of the individual components employed in a synthetic sequence to an appropriate solid phase support allows for the further creation of chemical diversity by utilizing not only parallel synthesis but also split synthesis wherein the solid 0 support containing the compounds prepared in the prior step can be split into a number of CK, batches, treated with the appropriate reagent and recombined.

The substrates can be attached to a solid support surface by any linkers known in the art. The linkers may be any component capable of being cleaved to release the substrate or final compound from the support.

Thus, libraries of compounds can be synthesized by initially attaching the first compound subst:rate to a solid support surface which can be performed by providing a plurality of solid support surfaces, suitably derivatizing each of the surfaces with groups capable of reacting with either the compound substrate or a linker moiety attached thereto.

The various support surfaces with the attached first compound substrate can then be subjected to various reaction conditions and second compound substrates to provide a library of attached compounds, which may, if necessary, be reacted further with third and subsequent compound substrates or varying reactions conditions. Attachment and detachment of substrates and products can be performed under conditions similar to those as described in Johnson, et al., Tetrahedron, 1999, 55, 11641; Han et al.

Tetrahedron 1999, 55, 11669; and Collini, et al., Tetrahedron Lett., 1997, 58, 7963.

Thus, the invention provides libraries of compounds of at least two compounds of Formula attached to a solid support surface or pluralities of surfaces.

Those skilled in the art will appreciate that the invention described herein in susceptible to variations and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications which fall P \operVnjl \I06 122M6 IN h )1 I Jo-2 I/I /1'2I o <U within the spirit and scope. The invention also includes all of the steps, features, compositions and compounds referred to or indicated in this specification, individually or c collectively, and any and all combinations of any two or more of said steps or features.

ID

00 Certain embodiments of the invention will now be described with reference to the 1^ 1 following examples which are intended for the purpose of illustration only and are not C1 intended to limit the scope of the generality hereinbefore described.

EXAMPLES

General methods: Melting points were recorded with a Kofler hot-stage apparatus and are uncorrected.

Proton and (1C) NMR spectra were recorded with a Varian Gemini 300 spectrometer operating at 300 MHz for proton and 75.5 MHz for carbon. All NMR spectra were recorded in (D)chloroform (CDCI 3 at 20 The protonicities of the carbon atoms observed in the carbon NMR were determined using attached proton test (APT) experiments. Infrared spectra (IR) we obtained as KBr discs or as films on NaCl plates and were recorded on a Perkin-Elmer Spectrum One Fourier-transform infrared spectrophotometer. Low-resolution electron impact mass spectra (MS) were recorded at eV on either a VG micromass 7070F instrument or a JEOL AX-505H mass spectrometer.

High-resolution mass spectra (HRMS) were recorded on a VG micromass 7070F instrument. Elemental analyses were performed on a Carlo Erba 1106. Tetrahydrofuran (THF) was distilled under nitrogen from sodium benzophenone ketyl. Dichloromethane was distilled from calcium hydride. Flash chromatography was performed on Merk Kieselgel P Oprnmal\20W6X 1 2264160 21 11 doc-2 /11/2I 6 0 -41- U INDOLES C OMe MeO OMe MeO N P" H C 6-Methoxy-2-(4-methoxyphenyl)-3-(3,4,5-trimethoxyphenyl)indole (BLF-36-1): 0 Methylmagnesium chloride (1.37 mL, 3.0 M in THF, 4.11 mmol) was added dropwise to a C solution of 2-iodo-5-methoxyacetanilide (550 mg, 2.0 mmol) and 4-methoxyphenyl acetylene (276 mg, 2.1 mmol) in dried THF (5.0 mL) at -5 The reaction mixture was then warmed to 18 Pd(PPh 3 2 C1 2 (42 mg, 0.06 mmol) added and the reaction mixture heated to 65 OC for 0.5 h, after which time the reaction was shown to be complete by The solution was cooled to 18 OC, diluted with DMSO (8.0 mL) and 3,4,5trimethoxyiodobenzene (617 mg, 2.1 mmol) added. The solution heated to 80 °C (external temperature) and a slight flow of N 2 for 1 h (to remove THF) and heating continued under a stationary atmosphere of N 2 for a further 10 h. The reaction mixture was cooled to 18 °C diluted with ethyl acetate (150 mL) and washed with water (2x 100 mL) and brine (3x 100 mL), dried over MgSO 4 and concentrated onto silica gel (5 g) under reduced pressure. The solid residue was subjected to flash chromatography (silica gel, eluted sequentially with hexane CH 2 C2 diethyl ether 2:1:1 and The relevant fractions 0.328, CH;:Cl 2 were concentrated giving the product as a white solid (782 mg, 82%).

'H NMR (300 MHz, CDCl 3 6 8.19 (br s, 1H), 7.60 J 8.7 Hz, 1H), 7.36 J 8.4 Hz, 2H), 6.91-6.82 4H), 6.65 2H), 3.92 3H), 3.87 3H), 3.82 3H), 3.74 (s, 6H). "C NMR APT (75 MHz, CDCI 3 6 158.9 156.6 153.1 136.4 136.2 132.8 130.9 129.2 125.2 123.0 120.0 113.9 (CH), 113.8 110.1 106.8 94.4 60.9 (CH 3 55.9 (2x CH 3 55.7 (CH 3 -1) 55.3 (CH 3 IR (KBr disc, cm 3368, 2999, 2933, 2831,1574, 1514, 1405, 1332, 1260, 1180, 1128. MS (70 eV) m/z 419 (M 100), 404 CH 3 P %apcrrn&)I12(i6%)I 2264 160) 2011 dc.; II I i2)66, -42- MeO

W

XOMe OiPr N- OMe MeO N

H

00 6-Methoxy-2-(3-isopropoxy-4-metoxypeny)-3-(3,4,5-trimthoxypbeyl)indole: Pd(PPh 3 2 C1 2 (11 mg, 0.0 15 mmol) and GuI (6.0 mg, 0.03 mmol) were added to a solution of triethylamine (140 LL, 1.0 mmol), 2-iodo-5-methoxytrifluoroacetanilide (173 mg, 0.50 mmol) and 3-.isopropoxy-4-methoxyethynylbeflzene (105 mg, 0.55 mmol) in dry c-I acetonitrile (4.0 mL). The reaction mixture was stirred at 18 'C for I h, after which time the reaction was shown to be complete by TLC. K 2 C0 3 (207 mg, 1.50 mmol) and 3,4,5trimethoxyiodobenzene 7 (162 mg, 0.55 mmol) were added, and the reaction mixture was stirred at 1 8 'C for 18 h. After this time it was diluted with diethyl ether (50 mL), washed with H 2 0 (2 x S0 mL), dried over MgSO 4 and concentrated onto silica gel (1 The solid residue was subjected to flash chromatography (silica gel, hexane CH 2

CI

2 diethyl ether 3:3: The relevant fractions (Rf =0.20, in eluant) were concentrated, giving the product as a white solid (183.2 mg, 77%) mp 184-5 0 C. 'H NMR (300 MHz, CDCI 3 6 8.54 (hr s, IlH), 7.54 J =8.7 Hz, I1H), 7.04 J =8.7 Hz, I 6.94 J 2.4 Hz, I 6.89 6.81 (mn, 3H), 6.68 2H), 4.24 (septet, J 6.0 Hz, I 3.90 3H), 3.84 6H), 3.75 (s, 6H), 1.20 J =6.0 Hz, 6H). 1 3 C NMR APT (75 MHz, CDCl 3 5 156.5 153.1 149.5 146.8 136.3 136.1 132.8 131.1 125.3 123.1 119.8 115.3 113.7 111.6 109.9 106.9 94.5 7 1.1 60.8 (CHA) 55.9 (CHA) 55.7 (CHA) 55.6 (CH 3 21.7 (CH 3 (2 x Ar CH superimposed). JR (KBr disc, cm. 3404, 3000, 2930, 2834, 1573, 1496, 1462, 1317, 1249, 1212, 1124. MS (70 eV) 477 100), 463 CH 3 13), 420 351 HRMS calcd for C 28

H

3 IN0 6 477.215 1. Found: 477.215 8.

P opcrVTall2Ul)6122( i) 2011 I doc 2111 1/21XI6 0 -43- U 6-Methoxy-2-(3-hydroxy-4-methoxyphenyl)-3-(3,4,5-trimethoxyphenyl)indole (BLF- 61-3): N Aluminium trichloride (80.0 mg, 0.60 mmol) was added to a solution of isopropylether of BLF-61-3 (above) (96.0 mg, 0.20 mmol) in dry dichloromethane (3 mL) and the reaction 00 mixture stirred at 18 OC for 2 h. The solution was then diluted with NH 4 Cl(aq) (sat., mL) and extracted with ethyl acetate (2 x 15 mL). The combined ethyl acetate extracts C1 were dried over MgSO 4 and concentrated onto silica gel (1 The solid residue was 0 subjected to flash chromatography (silica gel, hexane dichloromethane diethyl ether giving the product, BLF-61-3, as a white solid (81.0 mg, 93%) mp 98-9 OC. 'H NMR (300 MHz, D 6 DMSO) 6 11.15 1H), 9.03 1H), 7.43 J= 9.0 Hz, 1H), 6.89 (me, 4H), 6.68 (dd, J 2.1, 9.0 Hz, 1H), 6.57 2H), 3.77 3H), 3.76 3H), 3.69 (s, 3H), 3.64 6I). 3 C NMR APT (75 MHz, D 6 DMSO) 6 155.9 152.9 147.3 146.3 136.6 135.9 133.2 131.2 125.6 122.4 119.4 (CH), 115.6 11:2.3 112.2 109.7 107.0 94.5 60.3 (CH 3 55.8

(CH

3 55.7 (CH3), 55.4 (CH 3 IR (KBr disc, 3385, 2934, 2833, 1626, 1582, 1513, 1461, 1407, 1338, 1249, 1203, 1161, 1124. MS (70 eV) m/z 435 100), 420

CH

3 16). HRMS calcd for C 25

H

25 N0 6 435.1682. Found: 435.1681.

MeO OMe MeO 0 OiPr MN OMe MeO rN

H

6-Methoxy-2-(3-isopropoxy-4-methoxyphenyl)-3-(3,4,5-trimethoxybenzoyl)indole: Pd(PPh 3 2 C1 2 (1I mg, 0.015 mmol) and Cul (6.0 mg, 0.03 mmol) were added to a solution of triethylamine (140 iL, 1.0 mmol), 2-iodo-5-methoxytrifluoroacetanilide (173 mg, 0.50 mmol) and 3-isopropoxy-4-methoxyethynylbenzene (105 mg, 0.55 mmol) in dry acetonitrile (4.0 mL). The reaction mixture was stirred at 18 OC for 1 h, after which time the reaction was shown to be complete by TLC. K 2 C0 3 (207 mg, 1.50 mmol) and 3,4,5trimethoxyiodobenzene 7 (162 mg, 0.55 mmol) were added and the N 2 atmosphere exchanged for carbon monoxide (1 atm, balloon). This reaction mixture was stirred at 18 OC for 18 h. After this time it was diluted with diethyl ether (50 mL), washed with HzO (2 P %P&McJII2IX6U 2264 1 td) 21111 dcc.::1/1 112W6 -44x 50 mL), dried over MgSO 4 and concentrated onto silica gel (1 The solid residue was subjected to flash chromatography (silica gel, hexane CH 2

CI

2 diethyl ether 2:41 and The relevant fractions (Rf 0.50, hexane CH 2 Cl 2 diethyl ether 2:1:1) were concentrated, giving the product as a yellow solid (185.0 mg, 73%) mp 196-7 0 C. 'H 00 NMR (300 MH-Z, CDCI 3 5 9.74 (br s, IlH), 7.93 J =9.0 Hz, I 6.96 2H), 6.93 (me, 6.86 J 2.1 Hz, I 6.73 J1 1.8 Hz, I 6.62 J =8.4 Hz, I 4. (septet, J1= 6.0 H-z, IlH), 3.78 3H), 3.75 3H), 3.71 3H), 3.60 1. 13 J Hz, 6H). C NMR APT (75 MHz, CDCl 3 6 192.1 157.0 152.3 150.5 146.8 142.9) 140.9 136.5 134.5 124.6 123.0 122.1 (CH), 121.1 1 17.0 112.1 111.5 111.2 107.1 94.6 71.5 60.7 (CHA) 55.7 (CHA) 55.6 (CHA) 55.4 (CH 3 21.8 (CH 3 IR (KBr disc, cm 3344, 2939, 2835, 1614, 1575, 1541, 1492, 1459, 1420, 1335, 1256, 1203, 1125. MS eV) m/z 505 100), 463 CH 3

CH=CH

2 26), 308 218 HRMS calcd for C 29

H

3 IN0 7 505.210 1. Found: 505.2106.

Meo OMe MeO 0 OH -OMe MeO N

H

6-Methoxy-2-(3.*-hydroxy-4-methoxypbenyl)-3-(3,4,5-trimethoxybenzoyl)indole (BLF- 67-3): The isopropyl ether of BLF-67-3 (above) was cleaved as described for BLF-61-3 (above) (76.0 mg, 91%) mp =189-90 0 C. 'HNMR (300 MHz, CDCI 3 859.04 (br s, IH), 7.85 J 9.3 Hz, 1IH), 6.94 2H), 6.88 (in, 3H), 6.70 (dd, J =8.4 Hz, 1.8 Hz, I1H), 6.50 J= 8.4Hz), IH), (C,J=8.7 Hz, 1H), 6.95 1.5 Hz, IH), 6.88-6.82 3H), 5.69 lH), 3.82 3H), 3.79 3H), 3.75 3H), 3.67 6H). 1 3 C NMR APT (75 MHz, CDCI 3 8 192.5 156-9 152.3 146.9 145.2 143.1 140.8 136.4 134.9 124.9 122.8 122.0 121.4 114.9 112.4 107.1 94.5 60.7 (CHA) 55.9 (2x CH 3 55.7 (CM 3 55.5 (CH 3 IR (KBr disc, cm-1 3420, 3322, 2937, 2836, 1626, 1579, 1496, 1455, 1419, 1345, 1321, 1261, 1228, 1199, P Xap1rW A IU(11 6 122" 160l 20 11 dwol.2 11(X1A 1127. MS (70 eV) m/'z 463 100), 448 CH 3 10). HRMS calcd for

C

25

H

25 N0 7 463.163 1. Found 463.1648.

IND

00 P lOpor\,-JUQ$6ll 2264164) 2011 do.:.2111l1/2(XJ6 -46- BENZOFURANS AND BENZOPYRANS c-IMeO OMe MeO 08On 00 0 c-I 0 c-I2,3-1[2'-(3",4"-.methylened ioxypbenyl)-3'-(3" ',51"-trimcth oxyphenyl)fu ranoj1- 17-0benzylestradiol: This product was prepared as described for BLF-36-1 using 2-iodo-17-O-benzylestradiol (270 mg, 0.55 mmol), 3,4-(methylenedioxy)phenylacetylefle (104 mg, 0.71 mmol) and 3,4,5 -trim etho xyiodobenzene (211 mg, 0.72 mmol) giving white solid product (70% yield) 'H-NMR (CDCl 3 8 7.39 IH, Ar-H), 7.30 (in, 5H, Ar-H), 7.25 IH, Ar-H), 7.21 (dd, J 8.1 Hz and 1.8 Hz, I H, Ar-H), 7.16 J 1.8 Hz, I H, Ar-H), 6.78 J 8.1 Hz, I1H, Ar-H), 6.70 2H, Ar-H), 5.98 2H, O-CH- 2 4.59 2H, Ph-CH 2 3.97 3H, OCHA) 3.83 6H, 2x OCH 3 3.53 J 7.8 Hz, 1H, CH), 3.02 (in, 1H), 2.35 0.89 (I17H). 1 3 C-NMAR (CDCI 3 8 153.8, 152.3, 149.7, 147.7, 147.6 139.4, 137.4, 135.9, 134.4, 130.7, 128.7 128.3 (2 x Ar-CH), 127.4 (3 x ArCH), 124.9 121.2 (Ar-CH), 116.4 115.9, 110.5 108.5, 107.3 106.7 (2 x Ar-C), 101.3 (0-

CH

2 88.3 (CH-0-Bn), 71.7 (Ph-CH 2 61.1 (OCHA) 56.3 (2 x OCH 3 50.5, 44.5, (CH) 43.4 (CHA) 38.7 38.0, 30.2, 28.1, 27.4, 26.7, 23.3 (CHA) 11.9 (CH 3 LRMS m/z= 672 (100) 91 (26) (C 7 H7+).HRMS Calculated 672.308704. Found 672.309253.

IR (neat) v max =2932, 1580, 1505, 1489, 1465, 1453, 1411, 1379, 1299, 1236, 1128, 1105, 1038, 736.

Meo

OME!

MeO

X

MeO91 -a0 6- toy: t x hey)3(3 i t x e o n lblIfu ran (BLF-28-1): P %pDper% Ia ll 0 MI 22 I I I I I1 dm 2 1/ I1/2X

NO

0 -47- 0 q1 Methylmagnesium chloride (1.40 mL, 3.0 M in.THF, 4.2 mmol) was added dropwise to a solution of 2-iodo-5-methoxyphenol (500 mg, 2.0 mmol) and 4-methoxyphenyl acetylene

O

NC (290 mg, 2.2 rmol) in dried THF (5.0 mL) at -5 OC. The reaction mixture was then warmed to 18 Pd(PPh 3 2 Cl 2 (42 mg, 0.06 mmol) added and the reaction mixture heated 00 to 65 OC for 1 h, after which time the reaction was shown to be complete by The STHF was removed by passing a steady flow of N 2 over the heated solution. This was C cooled to 18 oC and DMSO (8.0 mL) added and the N 2 atmosphere exchanged for Scarbon monoxide (1 atm.) and stirred for 0.3 h. After this time 3,4,5- N trimethoxyiodobenzene (624 mg, 2.12 mmol) was added and the solution heated to 80 °C (external temperature) for 16 h. The reaction mixture was cooled to 18 OC diluted with ethyl acetate (100 mL) and washed with water (2x 80 mL) and brine (3x 80 mL), dried over MgSO 4 and concentrated onto silica gel (3 g) under reduced pressure. The solid residue was subjected to flash chromatography (silica gel, eluted sequentially with hexane

CH

2 C1 2 2:1, 1:1, The relevant fraction 0.23, hexane diethyl ether 2:1) were concentrated giving the product as a yellow solid (510 mg, mp 108-9 OC. 'H NMR (300 MHz, CDC1 3 8 7.56 J= 9.0 Hz, 2H), 7.53 J= 8.7 Hz, 1H), 7.13 2H), 7.09 J 2.4 Hz, 1H), 6.92 (dd, J 2.4 Hz, 8.7 Hz, 1H), 6.82 J 9.0 Hz, 2H), 3.89 (s, 3H), 3.87 3H), 3.79 3H), 3.70 6H). "C NMR APT (75 MHz, CDCI3) 6 190.4 160.2 158.2 156.9 154.4 152.5 142.0 132.3 129.5 122.0 121.5 121.4 114.2 113.5 112.3 107.0 (CH),

-I

95.2 60.6 (CH 3 55.7 (2x CH 3 55.3 (CH 3 54.9 (CH 3 IR (KBr disc, cm 2935, 2833, 1647, 1509, 1584, 1494, 1455, 1438, 1409, 1368, 1304, 1254. MS (70 eV) m/z 448 (M 100), 433 (M CH 3 15). HRMS calcd for C26H 24 07 448.1522. Found 448.1520.

MeO OMe MeO 0 OH MeO 0-6 OMe MeO 0 P op4V ,,.IA2I4)%I 22646IN 1201 doc.: ;1 12M -48- 6-Methoxy-2-(3-hydroxy-4-metboxyphenyl)-3(3,4,5trimnethoxyben2.oyl)benzo fu ran (BLF-62-3) The isopropyl ether of BLF-62-3 was prepared using an identical procedure as that described above for BLF-28-1 and the isopropyl ether group cleaved as described for IND BLF-61-3 above (60 over both steps). BLF-62-3: mp 68-9 0 C. 'H NMR (300 MHz, 00

CDCI

3 8 7.53 J =8.7 Hz, IRH), 7.24 J =2.1 Hz, I1H), 7.13 2H), 7.10-7.05 (in, Ni 2H), 6.91 (dd, J= 8.4 Hz, 2.1 Hz, I 6.72 J= 8.4 Hz, I 5.64 I1H), 3.89 3H), 3.87 3H), 3.86 3H), 3.71 6H). 1 3 C NMR APT (75 MHz, CDCI 3 8 190.9 158.4 156.9 154.6 152.7 147.6 145.4(C), 142.2 132.6 123.0 121.8 121.6 121.1 114.8 114.1 112.6 110.2 107.2 95.5 60.8 (CHA) 56.0 (2x CHO), 55.8 (CHO), 55.7 (CH 3 IR (KBr disc, cm l) 3400, 2940, 2837, 1622, 1581, 1494, 1462, 1414, 1262, 1232, 1126. MS eV) m/z 464 100), 449 CH 3 10), 408.2 (15) HRMS calcd for C 2 6

H

24 0 8 464.1471. Found 464.1459.

0 2-Pbenylbenzolblpyran-4-one.

n-Butyllithium (0.56 mL, 1.0 mmol) was added to a solution of 2-iodophenol (109.9 mg, 0.500 mmol) in THF (2 mL) at -78 0 C (dry-ice acetone bath). After 0.1 h, 3phenylpropynoyl chloride (82.2 mg, 0.500 mmol) was added, and the reaction warmed to room temperature. After I h, DMSO (4 mL) and water (0.5 mL) were added. After further stirring for I h, the solution was diluted with diethyl ether (50 mL), washed with NH4C(aq) 40 mL), wlater (40 mL), dried over MgSO 4 and concentrated under reduced pressure onto silica gel (2 The residue was subjected to flash chromatography (silica gel, hexane diethyl ether 9-1:5, 4: 1, then 1: 1) to give the product as a white solid (86 mg, 78%).

P ope'nlVl 21)6\122I,4~1N) 201 I doc-:; I/ Il/(I)t -49- 0 2-benzylidene-6-methoxybenzo[b]furan-3-one NO n-Butyllithium (0.86 mL, 1.72 mmol) was added to a solution of (200 mg, 0.858 mmol) in THF (4 mL) at -78 0 C (dry-ice acetone bath). After 0.15 h, 3-

(N

CN phenylpropynoyl chloride (142 mg, 0.858 mmol) was added, and the reaction warmed to O room temperature and quenched with NH 4 CI(aq) 30 mL). The solution was extracted C1 with diethyl ether (3 x 50 mL), and concentrated under reduced pressure to give a brown residue. The residue was diluted with methanol (6 mL), and AgNO 3 (90 mg, 0.515 mmol) added, and the solution stirred for 2 h. After this time, the solution was diluted with diethyl ether (50 mL), washed with water (40 mL), dried over MgSO 4 and concentrated under reduced pressure onto silica gel (2 The residue was subjected to flash chromatography (silica gel, hexane diethyl ether 9:1, 5:1, then 1:1) to give the product as a white solid (159, 74%).

INDANONES AND INDENONES MeOD -o -OMe MeO OMe 3-(3'-Isopropoxy-4'-methoxyphenyl)-l-(3",4",5"'-trimethoxyphenyl)propyn-l-one.

n-Butyllithium (9.4 mL, 2.00 M in hexanes, 19 mmol) was added dropwise to a solution /,/f-dibromo-3-isopropoxy-4-methoxystyrene (3.29 g, 9.39 mmol) in dry THF (50 mL) at 78 OC (dry ice/acetone) (produces lithium 3-isopropoxy-4-methoxyphenylacetylide). The solution was then allowed to return to room temperature before being again cooled to -78 OC. 3,4,5-Trimelhoxybenzoyl chloride [2.27 g, 9.86 mmol, dissolved in THF (20 mL)] was added and the solution once again allowed to warm to room temperature. The reaction mixture was diluted with diethyl ether (100 mL), washed with distilled water (2 x 50 mL), P NopcmalUM\l 2264 1 1011 dl2 .11I 1/2(16 0

O

0( dried over MgSO 4 and concentrated onto silica gel (1.5 g) under reduced pressure. The solid residue was subject to flash chromatography (silica gel, 2% ether in dichloromethane)

O

N giving a white solid (2.27 g, mp =130-31°C. 'H NMR (300 MHz, CDC1 3 6 7.50 2H, Ar-H), 7.28 (dd, J= 8.3Hz, 1.8Hz, 1H, Ar-H), 7.16 J= 1.8Hz, 1H, Ar-H), 6.89 oO J= 8.3Hz, 1H, Ar-H), 4.54 J= 6.1Hz, 1H, i-Pr), 3.96 6H, OMe), 3.94 3H, r"r OMe), 3.90 3H, OMe), 1.38 6H, J 6.1Hz, i-Pr).' 3 C NMR APT (75 MHz,

,I

SCDCl 3 6 176.8 153.0 152.9 147.1 143.3 132.3 127.3 (CH), S119.3 1:11.7 111.6 106.7 94.4 86.3.4 71.6 61.0

(CH

3 56.2 (CH 3 56.0 (CH 3 21.9 (CH 3 IR (KBr disc, cm 3011, 2976, 2938, 2838, 2187, 1637, 1586, 1510, 1460, 1414. MS (70 eV) m/z 384.1(M+), 342.0, 299.0, 175.0 OMe MeO OMe 0 MeOo MeO MeO OMe 2-(3'-Isopropoxy-4'-methoxybenzylidine)-1,3-bis-(3",4' trimethoxyphenyl)propan-l,3-dione.

Bis(dibenzylidineacetone)palladium (16 mg, 0.03 mmol) and triphenylphosphine (30 mg, 0.12 mmol) were dissolved in dry tetrahydrofuran (8 mL) under N 2 and stirred until the solution changed from red to yellow/orange (approximately 1 To this solution was added 1,3-diarylpropynone above (0.384g, 1.00 mmol), followed by dropwise addition of tributyltin hydride (0.28 mL, 1.0 mmol). The solution was then allowed to stir until TLC indicated complete reaction of the starting alkyne (approximately 0.5 after this time 3,4,5-trimethoxybenzoyl chloride (0.242 g, 1.05 mmol) and cuprous chloride (0.08 g, 0.80 mmol) were added. The solution was then stirred until TLC revealed complete consumption of the 3,4,5-trimethoxybenzoyl chloride (approximately 3 After this time the THF solution was taken up in diethyl ether (100 mL) and washed with aqueous KF solution 3 x 50 mL). The organic phase was then dried over MgSO4 and concentrated onto silica gel (1 g) under reduced pressure. The solid residue was subject to flash chromalography (silica gel, 4% diethyl ether in dichloromethane) giving a light P 64pm .1 2 4 )6)I2264 160) 2011 dm.211 11 421X)6 -51yellow solid (0.475 g, mp 105-6 'H NMR (300 MHz, CDCI 3 867.56 lH, 7.28 2H, Ar-H), 7. 10 2H, Ar-H), 6.97 (dd, J1= 8.5, 2.0 Hz, I1H, Ar-H), 6.86 J 2.0 Hz, I1H, Ar-H), 6.78 J] 8.5 Hz, I1H, Ar-H), 4.19 (in, J 6.1 Hz, I H, i-Pr), 3.91 3H, OMe), 3.88 3H, OMe), 3.83 9H, OMe), 3.80 6H, OMe), 1. 18 J= 00 6.1Hz, 6H, i-Pr). NMR APT (75 MHz, CDCI 3 8 196.0 193.4 153.2 c-i 152.9 152.4 147.1 143.3 143.2 141.8 136.7 132.6 c~K1 131.3 125.7 125.2 116.0 111.3 106.8 106.7(CH), 71.3 60.9 (Cl- 3 56.1 (CHA) 55.8 (CHA) 21.7 (CH 3 IR (KBr disc, cm )2971, 2940, 2836, 1646, 1578, 1504, 1122, 998. MIS (70 eV) m/z 580. 385.1, 327.0, 195.0, 152.0, 77.0.

MeO W~e MeO 0

W

MeO0 OMeg/ X We~ 3-(3'-isopropoxy-4'-methoxyphelyl)-4,5,6-trimethoxy- 2 3 trimethoxybeizoyl)-1-indalofe.

I ,3-Propadiont: above (0.581 g, 1.00 mmol) was dissolved in dry dichloromethane mL), to this solution was added methanesulfonic acid (68 j[tL, 1 .0 rmcl). After I h stirring at room temperature the solution was taken up in diethyl ether (50 mL) and washed with distilled water (2 x 20 mL), the organic phase was then dried over MgSO 4 and the solvent removed under reduced pressure, a light yellow solid was returned (0.576g, mp 65-6 0 C. The 'H NMR spectrum of this compound indicates the presence of an equilibrium mixture of the trans-isomer and an enol tautomer. 'H NMR (300 MHz, CDCl 3 5 7.26 7.19 7.05 6.83 6.79 6.68 6.67 6.64 6.61 6.56 5.14 4.99 J 2.5 Hz), 4.57 J =2.5 Hz), 4.42 4.29 3.71-3.95 (10 singlets), 3.45 3.32 1.31 1.26 1.20 1. 18 (d).MS (70 eV) m/z 580. 385. 1, P oM ,V&I 2(16%1122641602111 dcc.2111l112(X16 -52- 343.0, 195.0. IR (KBr disc, cm )2972 mn, 2937 mn, 2835 w, 1714 im, 1667 mn, 1583 s, 1505 s, 1336 s, 1125 vs MeO OMe I'D0 Me 00MeO

C

O~e 0 O~e 3-(3'-isop ropoxy-4'-metboxyphenyl)-4,5,6-trimethoxy-2-(3"'',4' trimethoxybenozoyl)indenone.

Indanone above (0.232 g, 0.40 mmol) and 2,3 -dichloro-5,6 -d icyanoqu inone (0.136 g, 0.60 mnmol) were dissolved in dry I ,2-dichloroethane (5 mL) and stirred at 60 'C for 3 days.

After this time the solution was decanted from the precipitate (dihydro-DDQ), concentrated onto silica gel (0.5 g) and chromnatographed (2:2:1 hexanes dichloromethane diethyl ether) to return a bright orange solid (185 mg, mnp 170-171 0 C. 'I- NMR (300 MF-z, CDC1 3 )8 7.16 (dd, J 2.1 Hz, I H, Ar-H), 7.11 I H, Ar-H), 7.03 2H, Ar-H), 7.02 J =2.1 Hz, I1H, Ar-H), 6.76 J 8.4 Hz, I H, Ar-H), 4.3 5 (in, J= 6.1 Hz, I H, i-Pr), 3.94 314, OMe), 3.92 3H, OMe), 3.84 3H, OMe), 3.82 3H, OMe), 3.77 6H, OMe), 3.39 3H, OMe), 1.25 J= 6.1 Hz, 614, i-Pr). 3 C NMR APT (75 MHz, CDCl 3 5 192.1 191.2 161.8 155.5 152.7 152.1 150.6 147.4 146.3 142.7 131.8 131.4 127.4 126.2, 125.0 122.2 116.0 110.5 106.9 105.0 71.5 61.6 (CH 3 61.2 (CH 3 60.8 (CHA) 56.5 (CHA) 56.1 (CH 3 55.8 (CH 3 21.9 (CH 3 IR (KBr disc, cm 3001, 2974, 2937, 2839, 1709, 1641, 1600, 1583,1507, 1460, 1416. MIS (70 eV) m/z 578.0(M+), 534.9, 368.9, 195.0 MeO OMe 0 OMe Meo- MeC Me/ HO e P \optrllmI26\)226aG 1)1 I doc-2 I 1/2(4)6 0 -53- U 3-(3'-Hydroxy-4'-methoxyphenyl)-4,5,6-trimethoxy-2-(3",4",5"- Strimethoxybenzoyl)indenone (DK-12a-1).

Indenone above (101 mg, 0.175 mmol) was dissolved in 5mL of dry dichloromethane, to this solution was added aluminium trichloride (94 mg, 0.700 mmol). After stirring for 0.15 oO h the mixture was taken up in diethyl ether (20 mL) and washed with aqueous ammonium chloride solution 2 x 30 mL), and water (20 mL). The organic phase was dried over CI MgSO 4 and evaporated onto silica gel (0.5g) under reduced pressure. The solid residue was O subject to flash chromatography (silca gel, 7% diethyl ether in dichloromethane) giving an N orange solid (71.4 mg, 'H NMR (300 MHz, CDC1 3 8 7.11 J= 2.1 Hz, 1H, Ar- 7.10 1H, Ar-H), 7.01 2H, Ar-H), 6.96 (dd, J= 7.8, 2.1 Hz, 1H, Ar-H), 6.67 J 7.8 Hz, 1H, Ar-H), 5.57 1H, OH), 3.94 3H, Ome), 3.91 3H, Ome), 3.85 3H, Ome), 3.84 Ome), 3.79 6H, Ome), 3.48 3H, Ome). 3 C NMR APT MHz, CDCl 3 6 192.1 191.0 162.1 155.7 152.7 150.7 148.1 147.4 144.8 142.5 132.0 131.5 127.3 126.2, 126.0 121.1 114.7 109.5 106.8 104.9 61.4 (CH 3 61.2 (CH 3

-I

60.8 (CH 3 56.6 (CH 3 56.1 (CH 3 55.8 (CH 3 IR (KBr disc, cm 3418, 2934, 2840, 1707, 1641, 1606, 1581, 1504, 1465, 1411, 1362, 1339, 1124. MS (70 eV) m/z 536.0(M+),493.0, 369.0, 343.0, 277.0, 219.0, 195.0 OMe MeO O Me MeOb MeO 0 OMe 3-(3'-Isopropoxy-4'methoxyphenyl)-l,2-bis(3",4",5 -trimethoxyphenyl)propen-1one: Bis(dibenzylidineacetone)palladium (13 mg, 0.024 mmol) and triphenylphosphine (24 mg, 0.10 mmol) were dissolved in dry tetrahydrofuran (8 mL)under N 2 and stirred until the solution changed from red to yellow/orange (approximately 1 To this solution was added propynone above (0.307 g, 0.800 mmol), followed by dropwise addition of P lopcPMIl'Xl\ 1 2,4IN)l 211 I/i 112K6 l0 -54tributyltin hydride (97% solution, 0.22 mL, 0.800 mmol). The solution was then allowed to stir for 5 hours, after this time 3,4,5-trimethoxyiodobenene (0.194 g, 0.84 mmol) and

O

CN cuprous chloride (0.16 g, 1.6 mmol) were added. The solution was then stirred until TLC revealed complete consumption of the 3,4,5-trimethoxyiodobenzene (3 days). After this 00 time the THF solution was taken up in diethyl ether (100 mL) and washed with aqueous KF solution 3 x 50 mL). The organic phase was then dried over MgSO 4 and CN evaporated under reduced pressure onto silica gel (1 Flash chromatography (sequential 0 elution 2% diethyl ether in dichloromethane) returned two isomers (higher Rf, 0.024 g, 5.4% lower Rf, 0.105 g, 23.7%).

Higher Rf isomer H NMR; 8 7.31 2H, Ar), 7.04 1H, 6.87 (dd, J= 8.2, 2.1Hz, 1H, Ar), 6.79 J= 2.1Hz, 1H, Ar), 6.73 J= 8.2, Hz, 1H, Ar), 6.64 2H, Ar), 4.22 J= 6.1 Hz, 1H, i-Pr), 3.88 3H, OMe), 3.85 3H, OMe), 3.84 6H, OMe), 3.80 9H, 2 x OMe), 1.20 6.1 Hz, 6H, i-Pr).

Lower Rf isomer 'H NMR; 8 7.26 1H, 7.09 2H, Ar), 6.83 (dd, J= 8.4, 2.1 Hz, 1H, Ar), 6.76 J= 8.4 Hz, IH, Ar), 6.63 J= 2.1, Hz, 1H, Ar), 6.51 2H, Ar), 4.04 J= 6.1 Hz, 1H, i-Pr), 3.92 3H, OMe), 3.86 9H, 2 x OMe), 3.83 3H, OMe), 3.77 6H, OMe), 1.18 J= 6.1 Hz, 6H, i-Pr).

APT 1 3 C NMR; 8 196.0 153.7 152.7 151.3 146.6 141.4 141.1 137.8 137.4 133.3 132.6 127.0 125.4 115.6 110.9 107.1 106.4 70.8 60.8 (CH 3 60.7 (CH 3 56.2 (CH 3 56.0 (CH 3 55.7 (CH 3 21.7 (CH 3 LRMS, m/z 552.2 509.2, 479.2, 345.1, 303.1, 269.0, 195.0 0 OMe MeO OMe MeO:

O

OMe O M e OiPr OMe P %OMpn I\ZI*)WI 22W 60 2011 doc2 III I/2(XI6 (±)trans-3-(3'-I sop ropoxy-4'-m ethoxyphenyl)-4,5,6-trimethoxy-2-(3' trimethoxypheiilyl)-1 -in da none: IN Prepared from 3-(3'-isopropoxy-4'methoxyphenyl)- I ,2-bis(3 trimethoxyphenyl)propen-1 -one (above) in a similar manner as described for 00 isopropoxy-4' -rnethoxyphenyl)-4,5 ,6-trimethoxy-2-(3" ,5 -trimethoxybenzoyl)- 1r- indanone (above~), giving the product as tan solid (80.8 mg, 'H NMR (300 MHz,

IN

IN CDCI 3 867.15 I H, Ar), 6.79 J =8.0 Hz, I1H, Ar), 6.63 (dd, J 8.0, 2.1 Hz, I H, Ar),

IN

6.61 J =2.1 Hz, I1H, Ar), 6.24 2H, Ar), 4.41 (in, J 6.1 Hz, I H, i-Pr), 4.41 J= (Ni 3.3 Hz, 1H, me-thine) 3.93 3H, OMe), 3.92 3H, OMe), 3.81 3H, OMe), 3.80 (s, 3H, OMe), 3.7.5 6H, OMe), 3.61 J= 3.3 Hz, I1H, methine) 3.3 8 3H, OMe), 1.30 J= 6.1 Hz, :3H, i-Pr), 1.26 J= 6.1 Hz, 3H, i-Pr). 13 C NMR APT (75 MHz, CDCI 3 6 204.5 155.1 153.4 150.2 149.3 149.1 147.1 143.0 137.0 135.9 134.7 131.5 119.8 115.4 111.9 104.9 100.8 71.3 65.0 60.8 (CHA) 60.7 (CHA) 60.1 (CHA) 56.2 (CH 3 56.0 (CHA) 55.9 (CHO), 22.0 (CHA) 21.8 (CH 3 0 OMe M e O MeOI'I e OWe WOe COMe (±)trans-3-(3'-1[sopropoxy-4'-metboxyphenyl)-4,5,6-trimethoxy-2-(3' ,4 trimethoxyphcnyl)-I -indenone (±)trans-3 sopropoxy-4 -methoxyphenyl)-4,5 ,6-trimethoxy-2-(3" trim ethoxyphernyl) -I-indanone (0.0801 g, 0.145 mmol) and DDQ (50 mg, 0.22 mmol) were dissolved in dry 1,2-dichloroethane (5 mL) and stirred at 80 'C for 12h. After this time the reaction mixture was evaporated onto silica gel (0.5 g) and flash chromatographed (7:7:1 hexanes dichloromethane diethyl ether) to return a red solid (42.6 mg, 'H NMR (300)MHz-, CDCl 3 8 7.06 1H, Ar), 7.02 (dd,J= 8.4, 1.9 Hz, I H, Ar), 6.91 (d, J= 1.9 Hz, 1H, Ar), 6.87 J= 8.4 Hz, IH, Ar), 6.44 2H, Ar), 4.37 (in, J= 6.1 Hz, IH, P ~cp~raIU(X)6\12 2.4 IN)2011 Ioc.21/11/l2(XI6 -56i-Pr), 3.91 3H, OMe), 3.88 3H, OMe), 3.87 3H, OMe), 3.78 3H, OMe), 3.62 6H, OMe), 3.33 3H, OMe), 1.24 J =6.1 Hz, 3M, i-Pr). 1 3 C NMR APT CK1MHz, CDC1 3 5 195.6 152.7 154.3 152.7 150.8 147.7 146.6 137.3 131.4 130.2 128.6 126.9 126.7 126.6 122.0 00 116.5 111.2 107.2 104.8 71.4 61.5 (CHO), 61.2 r-(CH 3 60.8 (CM 3 56.6 (CHA) 56.0 (CH 3 55.8 (CM 3 21.8 (Cl-I 3 THIOPHENES AND BENZOTHIOPHENES

PAC

4-[3'9-acetoxy -4'-methoxyphenyll-3-butylyl benzyl sulfide Cul (22 mg, 0.12 mmol) was added to a solution of benzyl 3-butynyl sulfide (502 mg, 2.85 mmol), 3 -ace toxy-4-methoxy-iodobelzene (643 mg, 2.20 mmol) and Pd(PPh 3 2 C1 2 (40 mg, 0.06 mmol) in a solution of DMF (3 mL) and Et 3 N (I mL). The resultant solution was stirred for 16 h at 18 The resultant solution was diluted diethyl ether (30 mL) and washed with HClaq in H 2 0, 30 mL) and water (3 x 30 mL), dried over MgSO4 and concentrated on to silica gel (2 g) under reduced pressure. The solid residue was subject to flash chromatography (silica gel, hexane diethyl ether 9: 1) giving the product as a viscous oil (734 mg, 'H NMR (300 MHz, CDCI 3 8 7.41 7.39 (mn, 5M), 7.28 (dd, J 2. 1, 8.4 Hz, IMH), 6.27 J =2.1 Hz, IMH), 6.87 J =8.4 Hz, IMH), 3.83 211), 3.82 3M), 2.68 2.32 3M). 1 3 C NMR APT (75 MHz, CDCl 3 8 168.4 150.9 139.2 138.0 130.2 128.7 128.3 126.8 125.8 115.8 112.0 87.2 80.4 55.7 (CM 3 36.1 (CM 2 30.1 (CM 2 20.5 (CM 2 20.3

(CM

3 IR (NaCl film, cm 3026, 2922, 2839, 1766, 1509, 1368, 1296, 1268, 1202, 1125.

MS (70 eV) m/z 340 22), 298 CM 2 CO, 65), 161 91 (100). HRMS calcd for C 2

(,H

2 0O 3 S 340.1133. Found 340.1129.

DAc 2-(39-aceto:y-49-methoxypheny)-4,5-dihydro-3-iodothiophene: P %qlrr"&1\2W6%122641N) 2011 dx.21/I 112M6 O -57- 0 Iodine (224 mg, 0.88 mmol) was added to a solution of benzyl 4-(3'-acetoxy-4'methoxyphenyl)-3-butynyl sulfide (300 mg, 0.88 mmol) in CH 2 C1 2 (3 mL) and the reaction C1 mixture stirred at 18 °C for 0.15 h. After this time the solution was diluted with diethyl ether (15 mL) and was washed with Na 2

S

2 0s w/v, 15 mL) and water (15 mL), dried

ID

oO over MgSO 4 and concentrated onto silica gel (1 The solid residue loaded onto a short column of silica gel (5 cm x 1.5 cm) and eluted sequently with hexane diethyl ether 9:1 CN and 8:2 giving the product as a white solid (328 mg, 99%) mp 81-3 'H NMR (300 0 MHz, CDC1 3 8 7.45 (dd, J= 2.1, 8.4 Hz, 1H), 7.29 J= 2.1 Hz, 1H), 6.95 J= 8.4 "1 Hz, 1H), 3.84 3H), 3.40 3.20 (me, 4H), 2.32 3H). "C NMR APT (75.5 MHz,

CDCI

3 6 168.6 151.1 138.9 138.9 127.4 127.1 123.3 (CH),

-I

111.6 73.6 55.8 (CH 3 49.4 (CH 2 32.0 (CH 2 20.5 (CH 3 IR (KBr disc, cm 2924, 2832, 1760, 1603, 1507, 1366, 1270, 1208. MS (70 eV) m/z 376 72), 334

CH

2 CO, 100), 161 HRMS calcd for C1 3

HI

3 0 3 SI 375.9630. Found 375.9633.

MeO OMe OMe OMe 2-(3'-acetoxy-4'-methoxyphenyl)-4,5-dihydro-3-(3",4",5",trimethoxyphenyl)thiophene (GPF-60-1): I-Butyllithium (0.36 mL, 1.7 M in hexanes, 0.61 mmol) was added to a solution 3,4,5trimethoxyiodobenzene (88 mg, 0.30 mmol) in THF (3 mL) at -78 oC (dry-ice acetone).

Zinc chloride (42 mg, 0.31 mmol) was added and the reaction mixture warmed to room temperature. Pd(PPh3) 2

CI

2 (7.0 mg, 0.01 mmol) and 2-(3'-acetoxy-4'-methoxyphenyl)- 4,5-dihydro-3-iodothiophene (77 mg, 0.20 mmol) were added and the resultant solution stirred at room temperature for 6 h. Methanol (2 mL) and K 2 CO3 (140 mg, 1.01 mmol) were added to the reaction mixture and stirring continued for a further 1 h. The reaction mixture was diluted with NH 4 CI(aq) (sat., 40 mL) and extracted with diethyl ether mL), dried over MgSO 4 and concentrated onto silica gel (1 The solid residue was subjected to flash chromatography (silica gel, hexane diethyl ether 2:1, 1:1) giving the product as white solid (63 mg, 'H NMR (300 MHz, CDC1 3 8 6.94 J 2.1 Hz, 1H), 6.80 (dd, J= 2.1, 8.4 Hz, 1H), 6.73 J= 8.4 Hz, 1H), 6.34 2H), 3.87 3H), P \oprVT n,l I\264(16)l 2(l11 dC-2 /1I 1/2(mI 0 -58- 3.83 3H), 3.64 6H), 3.34 (me, 4H), 2.32 3H). 1C NMR APT (75.5 MHz, CDC1 3 6 168.6 151.1 138.9 138.9 127.4 127.1 123.3 (CH), C 111.6 73.6 55.8 (CH 3 49.4 (CH 2 32.0 (CH 2 20.5 (CH 3 IR (KBr disc, cm 2924, 2832, 1760, 1603, 1507, 1366, 1270, 1208. MS (70 eV) m/z 376 72), 334 00 (M CH 2 CO, 100), 161 HRMS calcd for C1 3

HI

4 0 3 SI 375.9630. Found 375.9633.

OH

SS /OMe 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (272 mg, 1.2 mmol) was added to a solution of 2-(3'-acetoxy-4'-methoxyphenyl)-4,5-dihydro-3-iodothiophene (376 mg, 1.0 mmol) in dichloromethane (4 mL). After stirring at 18 °C for 1 h the reaction mixture was concentrated onto silica gel (2 g) and the solid residue subjected to flash chromatograph (silica gel, hexane diethyl ether 1:1) the product was obtained as a viscous resin (363 mg, This material was not fully characterized: 'H NMR (300 MHz, CDCI 3 7.48 (dd, J 2.1, 8.4 Hz, 1H), 7.35 J= 2.1 Hz, 1H), 7.26 J= 5.1 Hz, 1H), 7.12 J= 5.1 Hz, 1H), 7.03 8.4 Hz, 1H), 3.89 3H), 2.34 3H).

The resin was dissolved in methanol (6 mL) and K 2

CO

3 (690 mg, 5.0 mmol) added and the resultant slurry stirred at 18 OC for 1 h. The reaction mixture was then diluted with

NH

4 Cl(aq) (sat. 50 mL) and extracted with diethyl ether (2 x 50 mL). The combined organic fractions were dried over MgSO 4 and concentrated onto silica gel (2 The solid residue was subject to flash chromatography (silica gel, hexane diethyl ether 2:1, 1:1) and the product obtained as a viscous oil (302 mg, 91% from 2-(3'-acetoxy-4'methoxyphenyl)-4,5-dihydro-3-iodothiophene). 'H NMR (300 MHz, CDC1 3 8 7.25 J= 5.4 Hz, 1H), 7.22 J= 2.1 Hz, 1H), 7.15-7.10 2H), 6.92 J= 8.4 Hz, 1H), 5.77 (br s, 1H), 3.94 3H). "C NMR APT (75.5 MHz, CDC1 3 6 146.7 145.3 142.1 136.3 127.4 126.2 121.4 115.6 110.3 77.7

-I

55.9 (CH 3 IR (KBr disc, cm 3512, 2962, 2936, 2838, 1582, 1527, 1491, 1439, 1270, 1243, 1212, 1172, 1135, 1123. MS (70 eV) m/z 332 (M 100), 317 CH 3 58), 289 HRMS calcd for C IH 9 0 2 SI 331.9368. Found 331.9369.

P kopaw a 11X)(, 1226I616 20 1 dOm.2 1/ I1IXA

IND

\O

U MeO OMe MOMe D0 2-(3'-hydroxy-4'-methoxyphenyl)-3-(3",4",5",-trimethoxybenzoyl)thiophene

(BLF-

00 89-3): N t-Butyllithium (0.738 mL, 1.7 M in hexanes, 1.25 mmol) was added to a solution 3-iodo-2rC (3'-acetoxy-4'-methoxyphenyl)thiophene (139 mg, 0.418 mmol) in dry THF (4 mL) at -78 O °C (dry-ice acetone bath). To this was added a solution of 3,4,5-trimethoxybenzoyl chloride (108 mg, 0.47 mmol) in dry THF (1.5 mL) and the reaction mixture warmed to room temperature. The mixture was diluted with diethyl ether (50 mL) and washed with NH4Cl(aq)(sat., 50 mL), NaHCO3(aq) 60 mL) dried over MgSO 4 and concentrated onto silica gel (2 The residue was subject to flash chromatography (silca gel, hexane diethyl ether 4:1, 2:1, 1:1) and the product obtained as a white solid (109 mg, 65%) mp 151-2 OC. 'H NMR (300 MHz, CDCI 3 8 7.64 J 5.0 Hz, 1H), 7.29 7.24 2H), 7.22 (dd, J= 2.1, 8.4 Hz, 1H), 7.15 2H), 6.89 J= 8.4 Hz, 1H), 5.79 (br s, 1H), 3.94 6H), 3.92 6H). 13C NMR APT (75.5 MHz, CDCI 3 8 187.0 153.3 153.0 147.6 146.0 141.6 141.2 135.8 133.4 126.8 123.1 118.6 112.5 111.0 106.7 61.1 (CH 3 (56.4(CH 3 56.1

(CH

3 IR (KBr disc, cm 3212, 2993, 2939, 2839, 1574, 1453, 1428, 1348, 1259, 1240, 1126. MS (70 eV) m/z 400 100), 385 (M CH 3 12). HRMS calcd for

C

21

H

2 0 0 6 S 400.0981. Found 400.0982.

MeO SBn Benzyl 2-iodo-5-methoxyphenyl sulfide:

HBF

4 (50% w/v in H 2 0, 14 mL) was added to a stirred suspension of methoxyaniline 14 (5.00 g, 21.5 mmol) in H 2 0 (30 ml) and the suspension stirred at room temperature for 0.5 h. The resultant clear solution was cooled in an ice bath, giving a white suspension. To this suspension NaNO 2 (1.55 g, 22.5 mmol) in H 2 0 (10 mL) was added P %W I\2n6II 2264160 21111 I IUIX)6

IO

1) dropwise over 0.1 h and the reaction mixture warmed to room temperature. The resulting suspension was filtered, rinsed with water (50 mL) and diethyl ether (25 mL) and dried Cl under vacuum to give the corresponding diazonium tetrafluoroborate as a cream-colored solid 7.00 g (94 OO The diazonium salt (7.00 g, 20.1 mmol) obtained above was added portionwise to a solution of potassium ethyl xanthate (3.42 g, 21.0 mmol) in acetone (50 mL) at 0 OC (ice l bath) over 0.15 h. The reaction mixture was stirred at 0 °C for 0.75 h and at room O temperature for 1.0 h. This mixture was concentrated under reduced pressure diluted with Cl diethyl ether (50 mL) and washed sequentially with H 2 0 (50 mL), KOH (2 w/v in H 2 0, mL), brine (50 mL). The organic layer was died over MgSO 4 and concentrated under reduced pressure. The residue was dissolved in methanol (50 mL) and powdered KOH (3.38 g, 60 mmol) added and the reaction mixture stirred vigorously for 3 h. The methanol was then evaporated under reduced pressure. The residue was suspended in H 2 0 (40 mL) and CH 2

CI

2 (40 mL). Benzyl chloride (2.43 mL, 34.0 mmol) and n-Bu 4

NHSO

4 (100 mg) were added and the biphasic mix stirred vigorously for 1 h. The CH 2 C2 layer separated and the aqueous layer extracted with CH 2 C2 (50 mL). The combined CH 2 C2 fractions dried over MgSO 4 and concentrated on to silica gel (8 The solid residue was subjected to flash chromatography (silica gel, hexane diethyl ether 98:2) and the product was obtained as a colorless oil which crystallized upon standing at 4 °C to afford a cream solid (4.22 g, 59 (55 from 2-iodo-5-methoxyaniline) mp 72-4 'H NMR (300 MHz, CDC1 3 8 7.69 J= 8.7 Hz, 1H), 7.29-7.46 5H), 6.82 J= 3.0 Hz, 1H), 6.48 (dd, J= 3.0, 8.7 Hz, 1H), 4.16 2H), 3.70 3 'C NMR APT (75.5 MHz, CDC1 3 6 159.7 142.1 139.4 135.5 128.7 128.3 127.1 113.7 112.6

-I

87.4 55.0 (CH 3 38.5 (CH 2 IR (KBr disc, cm 2955, 2930, 1558, 1494, 1426, 1283, 1228, 1038. MS (70 eV) m/z 356 (M 45), 229 196 181 138 123 91 (100). HRMS calcd for C 14

HI

3 0SI 355.9732. Found 355.9728 OiPr MeOO OMe SBn Benzyl 2-[2'-(3"-isopropoxy-4"-methoxyphenyl)-ethynyl]-5-methoxyphenyl sulfide: P \o rimlUI21XH\1264 IN l)011 I dfoc-21/I I/2(XMI S-61- (1 n-Butyllithium (2.5 mL, 2.5 M in hexanes, 6.25 mmol) was added dropwise to a solution of ,f-dibromo-3-isopropoxy-4-methoxystyrene (1.09 g, 3.12 mmol) in THF (10 mL) at N78 °C (dry-ice acetone). After the addition was complete the cold bath was removed and the reaction mixture allowed to warm to room temperature over 0.33 h. Dry zinc chloride 00 (426 mg, 3.12 mmol) was then added and after it dissolved (approximately 3 min), Pd(PPh 3 2 C1 2 (35.0 mg, 0.05 mmol) and 2-iodo-5-methoxyphenyl sulfide (890 mg, 2.50 c mmol) were added. The resultant solution was stirred at room temperature for 1 h then Sdiluted with diethyl ether (30 mL) washed with NH4Cl(aq) (saturated solution in H 2 0, mL) and brine (30 mL) dried over MgSO 4 and concentrated onto silica gel The solid residue was subjected to flash chromatography (silica gel, hexane diethyl ether 9:1 then 3:1) to give the product (Rf= 0.25, 3:1) as a white solid (1.00 g, 96 mp 67-8 1

H

NMR (300 MHz, CDC1 3 5 7.43 J 8.4 Hz, 1 7.40-7.24 5H), 7.15 (dd, J= 1.8, 8.4 Hz, 1H), 7.09 J= 1.8 Hz, 1H), 6.84 J= 8.4 Hz, 1H), 6.79 J 2.4 Hz, 1H), 6.68 (dd, J= 2.4, 8.4 Hz, 1H), 4.55 (septet, J= 6.0 Hz, 1H), 4.23 2H), 3.88 3H), 3.75 3H), 1.38 J= 6.0 Hz, 6H). 13C NMR APT (75.5 MHz, CDC13) 5 159.5 150.8 146.9 141.1 136.8 133.6 129.0 128.6 127.3 (CH), 125.1 118.4 115.6 115.4 113.3 111.7 111.2 94.3 85.6 71.5 56.0 (CH 3 55.4 (CH 3 37.5 (CH2), 22.1 (CH 3 IR (KBr disc, -1 cm 2973, 2835, 1594, 1509, 1471, 1410, 1324, 1288, 1263, 1246, 1136, 1116, 1053. MS eV) m/z 418 100), 376 CH2=CHCH 3 34), 341 299 253 (58)91 HRMS calcd for C 26

H

26 0 3 S 418.1603. Found 418.1601.

I CiPr 2-(3'-Isopropoxy-4'-methoxyphenyl)-3-iodo-6-methoxybenzo[blthiophene: Iodine (556 mg, 2.19 mmol) was added to a solution of benzyl 2-[2'-(3"-isopropoxy-4"sulfide (900 mg, 2.15 mmol) in CH 2 Cl 2 mL) and the solution stirred at room temperature for 1 h. After this time the solution was washed with Na 2 S20 5 w/v, 30 mL), dried over MgSO 4 and concentrated onto silica gel (5 The solid residue loaded onto a short column of (5 cm x 2 cm) and eluted with hexane and hexane diethyl ether 3:1 to give the product (Rf= 0.33, as a white solid P lperlUlmaJ( n6\122641 60 2(l11 Jo- I/I 1/20U6 D -62- <d (950 mg, 97 mp 102-3 'H NMR (300 MHz, CDCI 3 5 7.69 J= 8.7 Hz, 1H), 7.30 J= 2.1 Hz, 1H), 7.26 J= 2.4 Hz, 1H), 7.21 (dd, J= 2.1 Hz, 8.4 Hz, 1H), 7.07 (dd, J 2.4, 8.7 Hz, 1H), 6.94 J 8.4 Hz, 1H), 4.63 (septet, J 6.3 Hz, 1H), 3.92 (s, 1 3H), 3.88 3H), 1.45 J= 6.3 Hz, 6H). "C NMR APT (75.5 MHz, CDC1 3 6 158.0 0. 150.5 146.7 139.5 139.3 135.9 126.9 126.5 122.6 116.7 115.1 111.4 104.3 (CH), N 77.7 71.3 55.9 (CH 3 55.6 (CH3), 22.1 (CH 3 IR (KBr disc, cm 2974, 2920, 0 2835, 1600, 1.530, 1493, 1471, 1261, 1224, 1138, 1020. MS (70 eV) m/z 454 66), 412 (M CH2=CHCH 3 58), 397 279 149 (100). Calcd for Cl 9

H

19 0 3 SI C: 50.23; H: 4.22. Found C: 50.27; H: 4.19.

MoO OMo MeO 0 OiPr -OMe 2-(3'-Isoprolpoxy-4'-methoxyphenyl)-6-methoxy-3-( 3 4 5 trimethoxybenzoyl)benzo[b]thiophene: t-Butyllithium (0.52 mL, 1.7 M in hexanes, 0.88 mmol) was added to a solution 3-iodo-2- (3'-isopropoxy- 4 '-methoxyphenyl)-6-methoxybenzo[b]thiophene (200 mg, 0.44 mmol) in dry THF (4 mL) at -78 °C (dry-ice acetone bath). To this was added a solution of 3,4,5trimethoxybenzoyl chloride (108 mg, 0.47 mmol) in dry THF (1.5 mL) and the reaction mixture wanned to room temperature. The mixture was diluted with diethyl ether (50 mL) and washed with NH 4 CI(aq) (sat., 50 mL), NaHCO3(aq) 60 mL) dried over MgSO 4 and concentrated onto silica gel (2 The residue was subject to flash chromatography (silca gel, hexane diethyl ether 4:1, 2:1, 1:1) and the product obtained as a colorless resin (200 mg, 'H NMR (300 MHz, CDC1 3 6 7.65 J 9.0 Hz, 1H), 7.32 J 2.1 Hz, 1H), 7.10 2H), 7.00 2H), 6.85 J= 2.1 Hz, 1H), 6.75 J= 8.4 Hz, 1H), 4.30 (septet, J= 6.0 Hz, 1H), 3.88 3H), 3.83 3H), 3.79 3H), 3.72 6H), 1.23 J Hz, 6H). "C NMR APT (75.5 MHz, CDC1 3 6 192.9 157.7 152.7 150.8 147.0 143.4 142.6 140.0 133.8 132.1 129.7 126.1 124.0 121.8 116.6 114.9 111.6 107.3 104.3 (CH), P lOPV" .I2[XW1l22b4160 211 IX I I 1/2(X)6 -63 71.5 60.8 (CHA) 56.0 (CHA) 55.8 (CHA) 55.5 (CHA) 21.8 (CH 3 IR (NaCi film, cm" '2936, 1644, 1581, 1531, 1501, 1473, 1413, 1228, 1126. MS (70 eV) m/z 522 ri 100), 480 CH2=CHCH 3 58), 301 195 HRMS calcd for C 2 9

H

3 0 0 7

S

522.1712. Found 522.1716 00 MeO OMe (N2~ MOO 0 "'\/OM0 MeOO S 2-(3'-Hyd roxy-4'-methoxyphenyl)-6-methoxy-3-(3",4",5"trimetboxybeu:toyl)benzo [bJ tb ophene (BLF-86- 1): Aluminium trichioride (86 mg, 0.64 mmol) was added to a solution of 2-(3'-Isopropoxy- 4' -methoxypheniyl)-6-methoxy-3-(3",4",5"-trimethoxybenzoyl)benzojb]thiophene (140 mg, 0.27 mmol) in dry dichloromethane (4 mL) and the solution stirred at room temperature for 1.5 h. After this time NH4C(aq) (sat., 20 mL) was added and the mixture extracted with dliethyl ether (20 mL) dried over MgSO 4 and concentrated onto silica gel (1 The residue was subject to flash chromatography (silica gel, hexane dichloromethane diethyl ether 3:3: 1) giving the product as a white solid (112 mg, mp 123-5 'H NMR (300 MHz, CDCl 3 867.67 J1=9.0Hz, I1H), 7.31 J1=2.4 Hz, I 7.06 2H), 7.00 (dd, J 9.0 Hz, I 6.98 J1=2.1 Hz, 1H), 6.83 (dd, J1=2.1, 9.0 Hz, I1H), 6.64 J1=9.0 Hz, 1 5.68 I 3.88 3H), 3.83 3H), 3.79 3H), 3.73 6H). 1 3

C

NMR APT ('75.5 MHz, CDC1 3 6 192.9 157.7 152.6 146.9 146.4 143.7 142 3 140.0 133.7 132.4 129.9 126.7 124.2 (CH), 121.3 115.1 114.9 110.4 107.3 60.8 (CHA) 56.0 (CHA) 55.8 (CHA) 55.5 (CH 3 IR (KBr disc, cm- 1 3402, 2934, 1649, 1580, 1499, 1474, 1413, 1324, 1266, 1228, 1158, 1125. MS (70 eV) m/z 480 100), 301 195 HRMS calcd for C 26

H

24 0 7 S 480.1243. Found 480.1242.

P %OPerVn I i i 20 I dOm-2I/1I/2M A

IO

-64- 0 OH

(N

00 0 S 2-(3'-Hydroxy-4'-methoxyphenyl)-5,6-methylendioxy-3-(3",4",5"rC trimethoxybenzoyl)benzo[b]thiophene (BLF-53-3) O Compound BLF-53-3 was prepared using a similar of reaction sequence as described for N BLF-86-1 giving the product as a white solid, mp 156-7 OC: 'H NMR (300 MHz, CDC1 3 8 7.24 1H), 7.21 1H), 7.05 2H), 6.95 J= 2.1 Hz, 1H), 6.80 (dd, J= 2.1, 8.4 Hz, 1H), 6.64 8.4 Hz, 1H), 6.03 2H), 5.56 1H), 3.82 3H), 3.82 (s, 3H), 3.75 6H). 3 C NMR APT (75.5 MHz, CDCl 3 8 192.9 152.7 147.3 147.0 146.9 145.5 144.6 142.4 134.4 132.5 (2x 130.1 126.9 121.3 115.1 110.5 107.4 102.5 101.5 (CH 2 101.1 60.9 (CH 3 56.1 (2x CH 3 55.9 (CH 3 IR (KBr disc, cm- 1 3418, 2939, 1628, 1582, 1501, 1465, 1335, 1279, 1232, 1124. MS (70 eV) m/z 494 (M 100), 480 301 (10)267 HRMS calcd for C 26

H

22 0 8 S 494.1035. Found 494.1038.

0 -Pro MeO&

B

2-Bromo-5-isopropoxy-4-methoxybenzaldehyde NBS (1.19 g, 6.70 mmol) was added to a solution of isopropyl isovanillin ether (1.24 g, 6.39 mmol), in DMF (6 mL) at room temperature, and heated to 80 oC. The reaction was monitored by TLC (eluent, hexane diethylether 1:1, product Rf= 0.63) and after 7 h was cooled to room temperature. The solution was diluted with diethylether (100 mL), washed with Na2S205 w/v, 100 mL), water (2 x 100 mL), dried over MgSO4, and concentrated under reduced pressure onto silica gel (5 The solid residue was subjected to flash chromatography (silica gel, hexane diethylether 95:5, then 9:1) to give the product (Rf 0.09, 95:5) as a white solid (1.65 g, mp 78-79 1

H-NMR

(CDC13) 8 10.16 1H, -CHO), 7.41 1H, Ar-H), 7.04 1H, Ar-H), 4.61 (septet, J= P I,,~AOU2~ I 2011 d-c. I I I112,X)6 1H, -OCH(CH3)2), 3.92 3H, -OCH3), 1.36 J1 6.0 1-Iz, 6H, -OCH(CH3)2).

13 C-NMR (CDCI13) 5 190.8 155.6, 147.1 126.4 (C-CHO), 120.0 (C-Br), 115.8, 113.5 71.5 (-CH(CH3)2), 56.4 (-OCH3), 21.8 (2x -CH(CH3)2). LRMS IND ~(Calculated for Cl I1H1303 8 1 Br): mlz =274 (2 1) 232 (100) CH2=CH-CH3).

00 HRMS Calculated for C II 1H1303 8 1 Br 274.0027. Found 274.0024. IR (KBr disc, cm- )v max =2971, 1681, 1587, 1508, 1432, 1386, 1268, 1217, 1158, 1109.

#PrO H MeO& SnH Benzyl 2-formyl-4-isopropoxy-5-metboxyphenyl sulfide Benzylmercaptan (1.20 mL, 10.2 mmol) was added to a stirring suspension of NaH (490 mg, 10.2 mnmol) in DMF at 0 *C (ice-bath). The reaction was left to stir at 0 'C until hydrogen evolut~ion had ceased. To this was added the bromobenzaldehyde above (2.78 g, 10.2 mmol), and the reaction was left to stir for 0.25 h then warmned to room temperature.

After 24 h, the reaction was diluted with diethylether (100 mnL), washed with HCI (40 mL, 1 NaOCI 40 mL) and water (3 x 100 mL), dried over MgSO4, and concentrated under reduced pressure onto silica gel (8 The solid residue was subjected to flash chromatography (silica gel, hexane diethylether to give the product (Rf 0. 16) as a yellow oil (2.87 g, 89%).

1 H-NMR (CDCl3) 8 10.21 IH, 7.35 1H, Ar-H), 7.22 7.09 (in, 5H, Ar-H), 6.77 I1H, Ar-H), 4.63 (septet, J =5.7 Hz, I1H, -OCH(CH3)2), 3.98 2H, 3.77 3H, -OCH3), 1.36 J 5.7 Hz, -OCH(CH3)2). 13

C-NMR

(CDC13) 8 190.6 154.5, 147.4 137.0, 132.2, 130.4 128.9, 128.4 (2x C- 127.3, 11-7.1, 113.1 71.2 (-CH(CH3)2), 56.1 (-OCH3), 41.7 (CH2), 21.8 (CH(CH3)2). LRMS m/z =316 (26) 274 (11) CH2=CH-CH3), 225 (17) (M+ CH2(C6H5)) 183 (59) CH2=CH-CH3, -CH2(C6H5)) HRMS Calculated for C18H2003S 316.1133. Found 316.1144. IR (NaCi film, cm- 1 v max 2988, 2861, 1666, 1578, 1495, 1439, 1388, 1349, 1334, 1262, 1158.

P op ,,I2,A 2 H I Id.c2 Ill /2( -66-

OH

,prO 0Mg 00 r- 1-(2'-Benzylthio-5'-isopropoxy-4'-methoxyphenyl)-3-(3' -isopropoxy-4"m ethoxypb enyl)p rop-2-yn-1 -oI n-Butyllithium (0.63 mL, 1.27 mmol) was added dropwise to a solution of/3,f-dibromo-3isopropoxy-4-raethoxystyrene 1 3 (221 mg, 0.633 mmol) in THF (1.5 mL) at -78 'C (dry-ice acetone). The solution was warmed to room temperature, then recooled to -78 'C and aldehyde abovie (190 mg, 0.601 mmol), added and left to stir for 0.25 h. The reaction was bought to room temperature again, quenched with NH4CI(aq) 40 mL), taken up into diethylether (40 mL), washed with water (30 mL), dried over MgSO4, and concentrated under reduced pressure onto silica gel (2 The solid residue was subjected to flash chromatography (silica gel, hexane diethylether 3:2) to give the product (Rf =0.16) as a yellow resin ('236.1 mg, IH-NMR (CDCI3) 8 7.30 IH, Ar-H), 7.25 7.09 (in, Ar-H), 7.01 (dd, J= 1.8, 8.3 Hz, IIH, 6.95 1.8 Hz, 1H, 6.77 (d,J= 8.3 Hz, I H, 6.75 I H, Ar-H), 6.07 J] 5.1 Hz, I H, 1 4.62 (septet, J1=6.1 Hz, IlH, -OCIH(CH3)2), 4.45 (septet, J1 6.1 Hz, I H, -OCH(CH3)2), 3.99 2H, -CH2), 3.83 3H, -OCH3), 3.70 3H, -OCH3), 2.40 J1= 5.1 Hz, I1H, I1-H), 1.38 (dd, 1 =3.7, 6.1 Hz, 6H, -C)CH(CH3)2), 1.33 J =6.1 Hz, 6H, -OCH(CH3)2). 13 C-NMR (CDCI3) 6 150.9, 149.7, 148.0, 146.8 138.2, 137.5 129.1, 128.5 (2 x 127.2, 125.2 123.0 119.0, 118.6 114.8 114.2, 111.6 87.9, 86.4 71.4, 71.3 63.0 56.0, 55.9 (-OCH3), 41.8 22.2, 22.0, 21.8 (2 x CH(CH3)2). LRMS m/z 506 (43) 415 (100) CH2C6H5). HRMS Calculated for C30H3405S 506.2126. Found 506.213 1. IR (NaCI film, cm- 1 v max 2978, 1592, 1504, 1386, 1264, 1110, 1044.

P 4WW4 0MMI2264 M11 dmoC2 II U21

IO

Me0 OPr OMe 00 5-Isopropoxy-2..(3'-isopropoxy-4'-methoxybenzoyl)-6-metboxybenzo[b]thiophene: N Iodine (418 mg, 1.65 mmol) was added to a solution of the alcohol above (835 mg, 1.65 r mmol) in dichlcromethane (5 mL) at room temperature. The reaction was monitored by 0 TLC (eluent hexane diethylether dichloromethane 4:3:3, product Rf= 0.53), and after 3 h the dichloromethane was removed under reduced pressure. The resultant red oil was taken up into diethylether (100 mL), and washed with Na2S205 100 mL), water (100 mL), dried over MgSO4, and concentrated under reduced pressure onto silica gel The solid residue was subjected to flash chromatography (silica gel, hexane diethylether 5:3) to give the product (Rf= 0.14) as a yellow oil (591 mg, 86%).

1

H-NMR

(CDCl3) 7.74 1H, Ar-H), 7.58 (dd, J= 2.0, 8.4 Hz, 1H, 7.49 J= 2.0 Hz, 1H, 7.29 1H, Ar-H), 7.28 1H, Ar-H), 6.95 J= 8.4 Hz, 1H, 4.60 (septet, J 6.0 Hz, 2H, 2x -OCH(CH3)2), 3.96 3H, -OCH3), 3.94 3H, -OCH3), 1.41 J= 2.4 Hz, 6H, -OCH(CH3)2), 1.39 J= 2.6 Hz, 6H, -OCH(CH3)2). 13 C-NMR (CDCI3) 8 187.7 153.9, 152.0, 147.0, 146.7, 141.2, 136.9, 132.6 131.3 130.6 123.8, 115.7, 110.5, 110.2, 103.8 71.6, 71.4 (CH(CH3)2), 56.1, 56.0 (-OCH3), 21.9, 21.8 (2 x CH(CH3)2). LRMS m/z 414 (59) 330 (100), 2 x CH2=CH- CH3). HRMS Calculated for C23H2605S 414.1500. Found 414.1493. IR (NaCl film, cm- 1 v max 2975, 1624, 1594, 1500, 1463, 1291, 1268, 1235, 1211, 1136.

HOo

OH

OMe 5-Hydroxy-2-(3'-ydroxy-4'-methoxybenzoyl)-6-methoxybenzobthiophene (KH-2-2) 5-Isopropoxy-2-(3'-isopropoxy-4'-methoxybenzoyl)-6-methoxybenzo[b]thiophene (above) was reacted with 5 equivalents of AIC 3 as described for BLF-86-1, giving the product, KH-2-2, in a 91% yield: P cpcr~nuI12l6%1 26460 21111 Oom.! It I 1121)06 -68- ()'I-I-NMR (CDC] 3 67.73 I1H), 7.50 J=2.1 Hz, I 7.47 (dd, J=2.1, 8.3 Hz), 7.29 I1H), 7.24 I 6.92 J =8.3 Hz), 5.78 I 5.75 111), 3.97 3H), 3.95 (s, N 3H).

1 3 C-NMR (CDC1 3 8 187.9, 150.1, 148.5, 145.3, 144.9, 141.6, 135.2, 133.2 131.5 (Coo 131.3 122.7, 115.5, 109.9, 109.1, 102.9 56.2, 55.1 (CH 3

'N.N

MOM0 1-(2'-Benzylthio-5'-isopropoxy-4'-metboxypbenyl)-3-(3' -isopropoxy-4' methoxyphenyl)prop-2-yn-1-one: Prepared by DI)Q oxidation of 1 -benzylthio-5 '-isopropoxy-4' -methoxyphenyl)-3-(3' isopropoxy-4"..methoxyphenyl)prop- 2 -yn-l-oI (1.2 equivalents of DDQ in CH 2

CI

2

OR

By reaction of benzyl (2-bromo-4-isopropoxy-5-methoxyphenyl) sulfide with one equivalent of nBuLi at -78 'C in THIF followed by addition of 3-(3'-isopropoxy-4'methoxyphenyl)propynoyl chloride (1.1 equivalents) 'H-NMR (CDCl1 3 8 7.91 1H), 7.45 7.21 (in, 6H), 7.13 J 1.6 Hz, IH), 6.86 J 8.3 Hz, I1H), 6.76 I 4.5 5 (septet, J 6.1 Hz, I 4.5 2 (septet, J 6.1 Hz, I1H), 4.17 2H), 3.87 3H), 3.75 3H), 1.40 J =6.1 Hz, 6H), 1.37 J =6.1 Hz, 6H).

3 C-NMR (CDCl 3 6 176:.0, 154.3, 152.6, 146.9, 143.4, 136.9, 136.4 128.7, 128.5, 127.2, 126.9, 126.7 120.9, 119.0 112.0 111.5, 109.2, 93.4, 86.8 71.9, 71.4 55.9, 55.8 (CHA) 37.3 (CH1 2 21.9, 21.8 (CH 3

OH

0M.

1-(2'-benzylttiio-5'-isopropoxy-4'-methoxypelyl)-1-(3' -trimetboxyphenyl)-3- '-isopropcixy-4' ''-methoxyphenyl)prop-2-yn-1-oI: P %upcr~r IUMM164 I~ W 2n I IdOC.21 1/ 2W,6 -69 z-BuLi (0.57 mL, 0.938 mmol) was added dropwise to a solution of iodo-3,4,5trimethoxybenzene (138 mg, 0.469 mmol) in THIF (3 mL) at -78'C (dry-ice acetone bath), and left to s-lir for 0.5 h. After this time, 1-(2'-benzylthio-5'-isopropoxy-4'methoxyphenyl)..3-(3 -isopropoxy-4' -methoxyphenyl)prop-2-yn- 1-one (237 mg, 0.469 00 mmol) was dissolved in THIF (2 mL) and added dropwise to the solution. After further stirring for 0.5 h, the solution was warmed to room temperature and quenched with

NH

4 C(aq) (40 mL, The solution was extracted with diethyl ether (3 x 50 mL), washed with waver (40 mL), dried over MgSO 4 and concentrated onto silica gel (2 The solid residue was subjected to flash chromatography (silica gel, hexanes diethyl ether, 1: 1, 2:3 sequential elution) to give the product (Rf =0.21, 3:2 diethyl ether: hexanes) as a yellow oil (277 mng, 8 'H-NMR (CDC1 3 5 7.23 7.20 (in, 3H), 7.14 I 7.10 7.05 (in, 3H), 6.99 J 2 Hz, 1H), 6.90 2H), 6.78 J= 8.4 Hz), 6.59 1H), 5.38 1H), 4.42 (septet, J =6.1 Hz, 2H), 3.87 2H), 3.86 3H), 3.84 3H), 3.81 6H), 3.59 3H), 1.35 J =6.1 Hz, 6H), 1.33 J= 6.1 Hz, 6H).

1 3 C-NMR (CDCI1 3 152.8, 151.1, 148.8, 146.9, 146.4, 140.6, 139.7, 137.4, 137.3 129.3, 128.4, 127.2, 125.2 122.5 120.7, 118.6, 115.8 114.6 111.6, 104.3 90.1, 88.0, 75.4 71.4, 71.2 60.8, 56.9, 55.9, 55.8 (CHA) 41.2 (CHA) 21.9, 21.8 (CH 3

M.O

.0M _0M M.0X

OM.

5-Isopropoxy-2-(3'-isopropoxy-4'-methoxybenzoyl)-6-mthoxy-3-(3' trimethoxyphenyl)benzolbltbiophene: Iodine (80 mg, 0.3 17 mmol) was added to a solution of 1-(2'-benzylthio-5'-isopropoxy-4'methoxyphenyl)- -trimethoxyphenyl)-3-(3' '-isopropoxy-4' P \oprmall\2(i06 12264 l)2l do21/ 1/2(X)6 0 methoxyphenyl)prop-2-yn-l-ol (208 mg, 0.309 mmol) in CH 2 Cl 2 (3 mL) and left to stir for h. After this time, the solution was quenched with Na 2 S20 5 (aq) (40 mL, and the N1 solution extracted with diethyl ether (3 x 50 mL), washed with water (100 mL), dried over MgSO 4 and concentrated under reduced pressure onto silica gel (2 The solid residue 00 was subjected to flash chromatography (silica gel, hexanes diethyl ether, 2:3 eluent) to r- give the product (Rf= 0.21) as a red solid (164 mg, 91%).

'H-NMR (CDCI 3 8 6.95 (dd, J= 1.9, 8.4 Hz, 1H), 6.91 J 1.9 Hz, 1H), 6.90 2H), S6.87 J 8.4 Hz, 1H), 6.73 1H), 6.56 1H), 4.53 (septet, J 6.0 Hz, 1H), 4.38 (N (septet, J= 6.0 Hz, 1H), 3.89 6H), 3.85 3H), 3.82 3H), 3.78 3H), 1.40 J= Hz, 6H), 1.32 J= 6.0 Hz, 6H) 0e40 M.0 Mooo O4 0

-OH

OMe 5-Hydroxy-2-(3'-hydroxy-4'-methoxybenzoyl)-6-methoxy-3-(3",4",5"trimethoxyphenyl)benzo[b]thiophene: Aluminium trichloride (17 mg, 0.138 mmol) was added to a solution of 5-Isopropoxy-2- (3'-isopropoxy-4'-methoxybenzoyl)-6-methoxy-3-(3",4",5"trimethoxyphenyl)benzo[b]thiophene (27 mg, 0.0466 mmol) in CH 2 C1 2 (3 mL) at room temperature and left to stir. After 1 h, a further amount of aluminium trichloride (17 mg, 0.138 mmol) was added. After a further 2 h, the solution was quenched with NH 4 Cl(aq) mL, extracted with ethyl acetate (3 x 50 mL), dried over MgSO 4 and concentrated under reduced pressure onto silica gel (2 The solid residue was subjected to flash chromatography (silica gel, hexanes diethyl ether, 4:1, 7:3, 3:2, 1:1 sequential elution) to give the product as a yellow solid (21 mg, 92%).

P \opcfV, aI\2(X)6%1 22&4 I6 W 20I11 do.2 1/1 I2X -71- 'H-NMR (CDC1 3 867.33 1H), 7.32 I1H), 7.19 (dd, J1=2.1, 8.3 H z, I1H), 7.1 J 2.1 Hz, I1H), 6.61 J =8.3 Hz, I 6.47 2H), 5.78 I1H), 5.42 I1H), 4.04 3H), NK 3.84 3H), 3.7:5 9H).

00 Omo 2-(ct-iodo-3'-iscopropoxy-4'-methoxybenzylidene)-5-isopropoxy-6methoxybenzolthiophen-3-one: Iodine (370 mg, 1.47 mmol) was added to a solution of 1-(2'-benzylthioxy-5'-isopropoxy- 4' -methoxypheriyl)-3-(3' -isopropoxy-4' -methoxyphenyl)prop-2-yn- 1-one (740 mg, 1.47 mmol) in CH 2 Cl 2 (20 mL) and stirred for 0.5 h. After this time, the solution was quenched with Na2S2O 5 (aq) (200 mL), extracted with diethyl ether (3 x 150 mL), dried over MgSO 4 and concentrated under reduced pressure onto silica gel (7 The solid residue was subjected to flash chromatography (silica gel, hexanes :diethyl ether, 3:2, 2:3 sequential elution), to give the product as a yellow oil (668 mg, 84%).

'H-NMR (CDC1 3 6 7.39 1H), 7.09 (dd, J1 2.1, 8.4 Hz, I 7.08 7.05 J 2. 1, 1H), 7.04 (dd, J 8.4 Hz, I 6.90 J =8.4 Hz), 6.96 J= 2.0 Hz, I1H), 6.83 J 8.4 Hz, I 6.80 1 6.75 I 4.63 4.46 (in, 4H), 3.95 3H), 3.90 (s, 6H), 3.85 3H), 1.42 1.35 (mn, 24H). 1 3 C-NMR (CDCl 3 6 178.9, 165.1, 157.2, 156.9, 151.5, 146.6, 146.3, 145.9, 139.3, 138.0, 137.7, 134.2, 133.6, 126.8, 124.9 122.1, 121.8, 116.2, 115.9 114.0 112.2, 111.4, 110.8, 110.5, 105.3, 103.8 98.7 71.6 56.5, 56.1 (CHA) 21.9, 21.8 (CH 3 P XomwaRI\2611\I2641NI 220 211 doc.2 i2(XI6 -72- UM.0O M M0 000 2- [a-(3"-isoprolpoxy-4"1-methoxypbenyl)-3' isop ropoxy-6-ni ethoxybenzolbl thiopben-3-one.

t-Butyllithium (0.35 mL, 0.55 mmol) was added dropwise to a solution of iodo-3,4,5trimethoxybenzene (80 mg, 0.27 mmol) in THIF (3 mL) at -78 'C (dry-ice acetone bath).

After 0.1 h, dry zinc chloride (36 mg, 0.27 mmol) was added and the reaction warmed to room temperature. After 0.25 h, Pd(PPh 3 2 Cl 2 (4 mg, was added, followed by iodoaurone (68 mg, 0.13 mmol). The reaction was left to stir for 18 h, after which the solution was diluted with diethyl ether (40 mL), washed with water (30 mL), dried over MgSO 4 and concentrated under reduced pressure onto silica gel (2 The residue was subjected to flash chromatography (silica gel, hexanes diethyl ether, 3:2, 2:3, 1:3 sequential elutions) to give the product as a white solid (52 mg, 'H-NMR (CDC1 3 8 7.30 I 7.28 I 6.97 (dd, J 8.4 Hz, IlH), 6.81 J =2.0 Hz, I 6.73 (d, J 8.4 Hz, I 4.5 5 (septet, J 6.1 Hz, I 4.39 (septet, J 6.1 Hz, IlH), 3.95 3H), 3.82 3H), 3.79 3H), 3.1 6H), 1.38 J =6.1 Hz, 6H), 1.24 J =6.1 Hz, 6H).

CM.

0 C.

.0S 0M.

OM.

2-12-Oxo-1 -(3'-isopropoxy-4'-methoxypbeny)-2-(3",4",5"-trimtoxypbenl).

etbylidenel-5.isopropoxy-6-metoxybenlZObI tbiopbeu-3-one.

P opv n&rVI(f 2164 160I 21I11 d-2 1, 1112(X) 0 -73-

O

0( t-Butyllithium (0.37 mL, 0.59 mmol) was added dropwise to a solution of iodo-3,4,5trimethoxybenzene (87 mg, 0.29 mmol) in THF (5 mL) at -78C (dry-ice acetone bath).

O

CN After 0.1 h, dry zinc chloride (40 mg, 0.29 mmol) was added and the reaction warmed to room temperature. After 0.25 h, Pd(PPh 3 2 C1 2 (4 mg, was added, and the nitrogen 00 atmosphere replaced with carbon monoxide. The solution was stirred vigorously, and after r-.

0.2 h iodoaurone (68 mg, 0.13 mmol) was added. The reaction was left to stir for 19 h, CN after which time it was diluted with diethyl ether (50 mL), washed with water (30 mL), Sdried over MgSO 4 and concentrated under reduced pressure onto silica gel (2 The residue was subjected to flash chromatography (silica gel, hexanes diethyl ether, 1:2, 1:3, 1:4 sequential elutions) to give the product as a white solid. 'H-NMR (CDCI 3 5 7.40 (s, 1H), 7.19 (dd, J 2.1, 8.3 Hz, 1H), 7.12 J 2.1 Hz, 1H), 7.03 1H), 6.88 2H), 6.61 J= 8.3 Hz, 1H), 4.62 (septet, J= 6.1 Hz, 1H), 4.58 (septet, J= 6.1 Hz, 1H), 3.91 3H), 3.81 6H), 3.79 6H), 1.38 J= 6.1 Hz, 6H), 1.33 J= 6.1 Hz, 6H).

Biological Activity A number of compounds according to the invention (Figure 11) were examined for their effects on tubulin polymerization, colchicine binding and growth of Burkitt lymphoma CA46 cells according to the procedures described in Verdier-Pinard el al., Mol.

Pharmacol., 1998, 53 62 and effects compared against Combretastatin A4(A) and benzothiophene(B) (see page The results are depicted in Table 1.

P %oa~s )6%12264J60 2011 do.2211 112(X)6

IN

00 74 Tabel 1: Effects of compounds on tubulin polymerisation, coichicine binding and growth of Burkitt lyniphoma CA46 cells (see FIGURE 11) Compound Inhibition of tubulin Inhibition of coichicine binding Inhibition of Polymerization' inhibit ion )b CA46 cell

IC

50 growth 5 jiM inhibitor 50 jAM IC5 0 (nM) inhibitor 2.1 1 91 ND I (I c) (B)c >0'-28 2000 (640c) BLF-86-1 3.5 0.3 6 61 500(52Ve) BLF-53-3 >0'2 31 >1000 GPF-60-1 3.6 1 64 88 390' BLF-89-3 1.0 0.1 67 -300c BLF-28-1 0.75 55 80 40(34') BLF-62-3 1.3± 0.2 80 -42' BLF-36-1 4 0 >1000 BLF-61-3 4.1± 0.6 28 -370e BLF-67-3 1.6 54 KH-2-2 2.4± 0.4 DK-12a-I 10-40 DK-2a-2 4-10 The tubulin concentration was 10 p±M. Inhibition of extent of assembly was the parameter measured.

tbThe tubulin concentration was 1.0 pM and the [3H]coichicine concentration was 5.0 PM.

c Data from Pinney K.G. et al Bioorg. Med. Chem. Let., 1999, 1081 and US Patent 5,886,025.

'The asterisk indicates that the rate but not the extent of assembly was inhibited by compound concentrations as high as ~The MCF-7 human breast carcinoma cell line was used.

Claims (15)

1. A compound of formula wherein X is 0; RIB-RID and R3A-R3E are independently selected from hydrogen, hydroxy, methoxy, and amino or any :2 adjacent Ri and/or R 3 groups from RIB-RID and R3A-R3E form a dioxolanyl group; R 2 is an optionally substituted aryl or optionally substituted heteroaryl group; A is a single bond, A' is C=O; and is an optional double bond.

2. A compound of formula according to claim 1 wherein represents a double bond.

3. A compound of formula (IV) P oper 2al21M61122M 160 2)1 doc-21Iti1I2lft \O -76- U R 3 X R 3 D R 3 8 IDR3E R3A 00 R 2 RS x R 7 R2E R2D wherein X is O, S or NR" is aryl, heteroaryl, aroyl, heteroaroyl, acyl, benzyl, alkyl, alkenyl, alkynyl or sulphonyl); A is a single bond, C=O, O, S or NR (R is hydrogen, C 1 6 alkyl, C 2 6 alkenyl, C2- 6 alkynyl, or C .7acyl); R2A-R2E and R3A-R3E are independently selected from hydrogen, hydroxy, optionally substituted alkoxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted acyloxy, amino, optionally substituted alkylamino, optionally substituted dialkylamino, optionally substituted acylamino or where any two adjacent R 2 and/or R 3 group from R2A-R2D and R3A-R3E together form -O-CH 2 and R 4 -R 7 are independently selected from hydrogen, hydroxy, alkoxy, alkyl, alkenyl, alkynyl, amino, alkylamino, dialkylamino, acyl, acylamino, heteroaryl and aryl.

4. A method of preparing a compound of formula R1A R 3 P \lpr\all2i21I 22M64 201)1 do.2111 I/2WIi \D C0 -77- U wherein X is O, NH or NR, (wherein R is H, sulfonyl, Ci. 6 alkyl, C 2 6 alkenyl, C2-6alkynyl, Ci1 7 acyl or an aryl or heteroaryl group); C A' is independently selected from a single bond or C=0; RIA-RID are independently selected from hydrogen, hydroxy, optionally substituted alkoxy, \0 00 optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted acyloxy, amino, optionally substituted alkylamino, optionally C1 substituted dialkylamino, optionally substituted acylamino, or any 2 adjacent RIA-RID 0 together form -O-CH 2 c R 2 and R 3 are independently optionally substituted aryl or optionally substituted heteroaryl groups; and is an optional double bond; said method comprising the steps of: a) coupling a compound of formula with an alkyne of formula in the presence of a nickel or palladium coupling agent R1A R 1 B H a l M2 R2 Ric T XM 1 RID (2) wherein RIA-RID, R 2 and X are as above; Hal is I ,Br or Cl; MI is a metal or a metal species thereof, said metal selected from the group consisting of Li, Na, K, Mg, Cs and Ba; M 2 is a metal, or a metal species thereof, said metal selected from the group consisting of Mg, Zn, Cu, B, Si, Mn, Sn, Ge and Al; and P %OpvUn,%ld O\l22416112011 do 2111 12(X)6 IO -78- (U X is O or NR (R is sulfonyl, C 16alkyl, C 2 6 alkenyl, C2- 6 alkynyl, or C 7 acyl); Sb) reacting in situ the resulting coupled product with R 3 -L wherein R 3 is an optionally substituted aryl or optionally substituted heteroaryl group, and wherein L is a leaving 00 group, optionally in the presence of carbon monoxide. N'

5. A method for preparing a compound of formula C' R1A R3 R1B A Ric X R2 Rio wherein X is S; A' is selected from a single bond or C=0; RIA-RID are independently selected from hydrogen, hydroxy, optionally substituted alkoxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted acyloxy, amino, optionally substituted alkylamino, optionally substituted dialkylamino, optionally substituted acylamino, or any two adjacent RIA-RID together form -O-CH 2 and R 2 and R 3 are independently optionally substituted aryl or optionally substituted heteroaryl groups; said method comprising the steps of: a) coupling a compound of formula with a compound of formula in the presence of a nickel or palladium coupling agent P \opVnll2(I)M2\26A1N 6I21111 doc-2 I 'l2111)6 -79- wherein RIA-RIC, Hal, M 2 and R 2 are as above, and Ps is a sulfur protecting group capable of stabilizing a positive charge; b) cyclising the resulting coupled product in the presence of a Hal' producing reagent to give R1A Hal SR2 Ric S RlD wherein Hal is Cl, Br or I; c) coupling with either the moiety R 3 or R 3 wherein R3 is an optionally substituted aryl or optionally substituted heteroaryl group.

6. A method for preparing a compound of formula P \openMalUW\(MI:264160 2s111 doc-21,1 I /2t6 ID 0 SR1A (N 01 RID O wherein A is selected from a single bond or C=0; RIA-RID are independently selected from hydrogen, hydroxy, optionally substituted alkoxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted acyloxy, amino, optionally substituted alkylamino, optionally substituted dialkylamino or any 2 adjacent RIA-RID together form -O-CH 2 is an optional double bond; and R 2 and R 3 are independently optionally substituted aryl or optionally substituted heteroaryl groups; said method comprising the steps of: reacting compound with compound or reacting compound 6(a) with compound 7(a); P %op6,6041\21N61226416 2011 doc.2 1/11/26666 IND -81 D 0 M R (7) Rj 10 (6a) (7a) to form a compound (9) P O, r lIaXl2\ 12264 160) 211 6Cdc2 /1 11/2( -82- wherein M is Li, Na, K or MgHal (Hal is Br, Cl or I); b) agent; treating compound with a metal hydride in the presence of a palladium coupling c) coupling the resulting product with R 3 -Hal or R 3 -C(O)-Hal (wherein Hal is Cl, Br or I) to provide either compound (10) or and R3 RRi 0 Ro O (11) d) cyclising (10) or (11) under acidic conditions to form an indanone and optionally treating the cyclised product with an oxidising agent to form an indenone.

7. A method for preparing a compound of Formula RID wherein X is O, S or NR (wherein R=H, Ci. 6 alkyl, C2 6 alkenyl, C 2 6 alkynyl, or C(O)CI. 6 alkyl); P \oprnal l\X)6\12264160 2(11 dOc-:'ll 1/2(116 -83- RIA-RID are independently selected from hydrogen, hydroxy, optionally substituted alkoxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted acyloxy, amino, optionally substituted alkylamino, optionally substituted dialkylamino or any 2 adjacent RIA-RID together form -O-CH 2 A is C=0; R 2 and R 3 independently are optionally substituted aryl or optionally substituted heteroaryl groups; and is an optional double bond; comprising the steps of a) coupling a compound (12) with compound (13) RlA RIB Hal C XP Ric R1D (12) wherein Hal is CI, Br, or I; 0 Hal R2 (13) P \opa f.4 64XX\61 2(IM I I)20 dcc.2I II/2(fi -84- to form a compound of formula (14); R1A 0 RIB I R 2 Ric XP R1D (14) where X is S, protecting the thiol with a sulfur-protecting group b) reacting (14) with a compound M1---R 3 wherein MI is Li, Na, K, Mg, Cs or Ba, and R 3 is an optionally substituted aryl or optionally substituted heteroaryl group; to form R1A HO R3 RlB I R2 RI c XP RID wherein when X is O then P is H and when X is S, P is a sulfur protecting group and when X is NR, R is a hydrogen, sulfonyl, Ci-6alkyl,C 2 6 alkenyl, C2. 6 alkynyl, Ci-7acyl, or an aryl or heteroaryl group; c) treating (15) with a Hal producing reagent, to afford cyclisation. P 2 1 1 M do QW '(6

8. A compound of Formula (III) 00 R 5 A R 6 SR wherein R 2 and R 3 are independently optionally substituted aryl or optionally substituted heteroaryl groups; N'is CO; R 5 and R 6 can independently be hydrogen, optionally substituted alkyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl or optionally subsiituted alkenyl; and is an optional double bond.

9. A compound selected from the group consisting of: 6 -mnethoxy-2 -(4--methoxyphenyl1)- 3-(3 ,4,5 -trim ethoxyphe ny1) indo le; 6-methoxy-2-(3 ;-isopropoxy-4-methoxyphenyl)-3 -(3,4,5-trimethoxyphenyl)indole; 6-m ethoxy-2 isopropoxy-4-methoxyphenyl)-3 -trim ethoxybenzoyl) indol e; 6-methoxy-2-(3 '-hydroxy-4-methoxyphenyl)-3 ",4"-nmethylenedioxyphenyl)-3'-(3"' ,5"'-trimethoxyphenyl)furano]- 17-0- benzylestradiol; 6-methoxy-2-(4I'-methoxyphenyl)-3-(3" "-trimethoxybenzoyl)benzo[b] furan; 6-methoxy-2-(:3-hydroxy-4-methoxyphenyl)-3 ,4,5-trimethoxybenzoyl)benzo furan; 3 isopropox:y-4'-methoxyphenyl)-4,5 ,6-trim ethoxy-2 ,5"-trim etho xybenzoylI)- I indanone; P opcrV,,.Ik2(X)6X22(A 1691 2011 dC. III I 1(I6 -86- (1)3 -(3'-isopropoxy-4'-methoxyphenyl)-4,5 ,6-trimethoxy-2-(3 ,5 trimethoxybenzoyl)indenone; N3-(3'-hydroxy-4.'-methoxyphenyl)-4,5 ,6-trimethoxy-2-(3 ,5 IND trimethoxyben2zoyl)indenone; 00 (±)trans-3 -(3'-is;opropoxy-4'-methoxyphenyl)-4,5,6-trimethoxy-2-(3" t,4" ,5 trimethoxypheriyl)- 1 -indanone; (±)trans-3 -(3'-i!sopropoxy-4'-methoxyphenyl)-4,5,6-trimethoxy-2-(3" trimethoxyphenyl)- 1 indenone; 2-(3'-acetoxy-4 '-methoxyphenyl)-4,5-dihydro-3-(3" ,14",5 ",-trimethoxyphenyl)thiophene; 2-(3'-hydroxy-:3'-methoxyphenyl)-3 ",-trimethoxybenzoyl)thiophene; 2-(3'-isopropox.y-4'-methoxyphenyl)-6-methoxy-3 trimethoxyben:zoyl)benzo [bljthiophene; 2-(3 '-hydroxy-,4'-methoxyphenyl)-5 ,6-methylendioxy-3 trim ethoxybenzoyl)benzo thiophene; 5-isopropoxy-2 -(3'-isopropoxy-4'-methoxybenzoy)-6-methoxybenlzo[b]thiophene; -hydroxy-2-(''hydroxy-4'-methoxybelzoyl)-6-methoxybenzo[blthiophele; 5-isopropoxy-2Z-(3'-isopropoxy-4'-methoxybenzoy1)-6-methoxy- 3 3 '1,411,5'- trimethoxyphenyl)benzolthiophene; 5-hydroxy-2-(*3 '-hydroxy-4'-methoxybenzoyl)-6-methoxy-3 ,5 trimethoxyphenyl)benzo[blthiophene; "-isopropoxy-4"-methoxyphenyl)-3 '-trimethoxybenzylidene]-5-isopropoxy- 6 methoxybenzo [bllthiophen-3 -one; 2-[2-oxo- 1 isopropoxy-4'-methoxypheny l)-2 3 5 %trim ethoxyphenylI)-ethyl i dene] -i sopropoxy-6-methoxybenzo[b]thiophel-3 -one.

Use of a compound according to claim 9 in the manufacture of a medicament to treat conditions which require an anti-mitotic agent.

11. Use according to claim 10 wherein the condition to treat is a tumour.

12. A method of treating a condition requiring an anti-mitotic agent including administering to a subject in need thereof a compound according to claim 9. 01-04-'08 16:21 FROM- T-512 P011/014 F-140 I lWlJA,.;llif 64INJ2 ]pWd.flc.;1ItlnRI 00 -87- 0 0

13, A method according to claim 12 wherein the condition to be treated is a tumour.

14. A composition comprising a compound according to claim 9 together with a O pharmaceutically acceptable carrier. 00

15, A kit when used in screening compounds for TPI activity said kit comprising at least one substrate from each of group group and group S(a) R1A A RiA (ai) R a H l G R"l O I Hal (i) (ii) (iii) R2 or a metallated form thereof; or R2-C(O) -Hal; or PsS' 3 or a metaUated form thereof wherein L is replaced by a metal; or (ii) Rs-C(O)- Hal; wherein COMS ID No: ARCS-184998 Received by IP Australia: Time 16:22 Date 2008-04-01 P optaI\U20061I22 6 416I 2011 doc-19112f006 S-88- 0 RIA-RID are independently selected from hydrogen, hydroxy, optionally substituted alkyl, Soptionally substituted alkenyl, optionally substituted alkynyl, optionally substituted O C alkoxy, optionally substituted acyloxy, amino, optionally substituted alkylamino, optionally substituted dialkylamino or any 2 adjacent RIA-RID together form oO -O-CH2-O-; Hal is I, Br or Cl; Ci X' is OH, SlPs (wherein Ps is a sulfur-protecting group capable of stabilising a positive 0 charge), NPN (wherein PN is a nitrogen-protecting group); or NHR (wherein R is CI sulphonyl, triflouroacyl, CI.7acyl or an aryl group); L is a leaving group; and R 2 and R 3 are optionally substituted aryl or optionally substituted heteroaryl groups.