AU749165B2 - Preparation of fused polycyclic alkaloids by ring closure of azomethine ylides, novel compounds thereof and their use as chemotherapeutic agents - Google Patents

Description

WO 98/50365 PCT/AU98/00312 -1- PREPARATION OF FUSED POLYCYCLIC ALKALOIDS BY RING CLOSURE OF AZOMETHINE YLIDES. NOVEL COMPOUNDS THEREOF AND THEIR USE AS CHEMOTHERAPEUTIC AGENTS FIELD OF THE INVENTION The present invention is generally directed to a method for preparing compounds useful in therapy. More particularly, the present invention provides a method for preparing a class of fused polycyclic alkaloids as well as novel compounds obtained thereby, pharmaceutical compositions containing them and methods of treatment using them.

BACKGROUND OF THE INVENTION Naturally occurring molecules which exhibit potentially beneficial pharmacological properties are isolable from a range of environments, such as marine, plant and microbial sources. One example of such molecules is the general class of compounds known as the Lamellarins. These polyaromatic alkaloids are isolated from marine sources and comprise a fused polyaromatic framework. Lamellarins C and D have been shown to cause inhibition of cell division in a fertilised sea urchin assay, whereas Lamellarins I, K and L all exhibit comparable and significant cytotoxicity against P388 and A549 cell lines in culture. Recently, Lamellarin N has been shown to exhibit activity in lung cancer cell lines by acting as a Type IV microtubule poison. Furthermore, these compounds have also been shown to possess cytotoxic activity on multidrug resistant cells as well as efficacy as non-toxic modulators of the multidrug resistant phenotype and, therefore, afford an attractive potential source of chemotherapeutic agents.

However, the potential clinical usefulness of the Lamellarins is severely limited by the modest quantities produced naturally as well as the difficulties involved in their isolation.

Steglich coworkers, inAngew. Chem. Int. Ed. Eng. 1997, 36, 155, have described a biomimetic sequence for the synthesis of Lamellarin G trimethyl ether, however, the process is limited in that it lacks regiochemical control and does not readily lend itself to the specific substitution patterns dictated by the natural products. There is a need, WO 98/50365 PCT/AU98/00312 -2therefore, for a synthetic process which enables the production of the Lamellarins and analogues thereof.

SUMMARY OF THE INVENTION Throughout this specification, 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 group of integers but not the exclusion of any other integer or group of integers.

In a first aspect, the present invention contemplates a method for the preparation of a compound of general Formula comprising the step of cyclizing an azomethine ylide of general Formula (II): Zn -0 x ^©0 w wherein, -3- A is a-cyclic group being an optionally substituted aryl group or an aromatic heterocyclic group; or A is a cyclic group RAlRA 2 C-CRARA4 wherein RA2 and RA, together with the carbon atoms to which they are attached form an optionally substituted saturated or unsaturated caibocyclic or heterocyclic group and RA1 and RA 4 are as defined below or together form a bond; or A is a non-cyclic group RA1R4C-CRRA 4 wherein RA 1

RA

4 are as defined below and R A 2 and R may optionally together form a bond; Z is a carbon or a heteroatom; n is selected from 0, 1, 2 or 3; and

R

AIA4 and Y may be the same or different and each are selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally :substituted alkynyl, optionally protected hydroxy, optionally substituted amino, optionally substituted alkoxy, optionally substitutedalkenoxy, optionally substituted alkynoxy, optionally substituted aryl, optionally substituted heterocyclyl, carboxy, carboxy ester, carboxamido, acyl, acyloxy, mercapto, optionally substituted alkylthio, halogen, nitro, sulfate, phosphate and cyano, and W and X, together with the nitrogen and carbon atoms to which they are attached, form an isoquinolinyl or dihydroisoquinolinyl group which may S be optionally substituted; 20 or pharmaceutically acceptable derivatives and salts, racemates, isomers and/or tautomers thereof.

Another aspect of the invention contemplates a compound of Formula prepared by the methods as described herein.

Yet another aspect of the invention relates to novel compounds of general Formula (I) Q:operr\mjc2239249 April.doc-049/4)2 -4wherein A, Z, W, X, Y and n are as defined above, provided the compound is not one selected from Lamellarins A to N and S to X, Lamellarin I-acetate, Lamellarin B, C and Gdiacetates, Lamellarin A, D, E, G, K, L, M and N-triacetates, Lamellarin H-hexaacetate, Lamellarin T, U, V and Y-20-sulphate, Lamellarin G-trimethyl ether and Lamellarin Imethylate.

Still yet another aspect of the present invention relates to a method of treating multidrug resistant tumours comprising the administration of an effective amount of a compound of Formula A further aspect of the invention provides compositions comprising a compound of Formula together with a pharmaceutically acceptable carrier, excipient or diluent.

DESCRIPTION OF THE PREFERRED EMBODIMENTS The azomethine ylides of general Formula (II) are obtainable from corresponding precursors by methods known to those skilled in the art, for example as described by A.

Padwa et al, in Chem Rev., 1996, 96, 241 and V.P. Litvinov, Russian Journal of Organic Chemistry, Vol. 31 (No. 10), 1995, pp. 1301-1340. A particularly suitable means of generating the azomethine ylide of general Formula (II) is effected by the addition of a base to a compound of Formula (III).

WO 98/50365 PCT/AU98/00312 Zn wL

W

(II)

wherein the counter ion LO is a stable, weakly basic anion.

Suitable anions include those derived from the sulfonates, such as, tosylate, mesylate, triflate, bosylate, besylate, tresylate, nonaflate and nosylate and the halogens, especially chlorine and bromine and iodine. Preferably L' is bromide or iodide.

In a preferred embodiment, the present invention provides a method for the preparation of a compound of Formula (Ia): (Ia) comprising the step of cyclizing an azomethine ylide of general Formula (IIa): WO 98/50365 PCT/AU98/00312 -6-

Y

(IIa) wherein R'-R 8 and R 1 4 are as defined for W, X and Y as described above.

Preferably, the azomethine ylide of Formula (IIa) is generated by the addition of a base to a compound of general Formula (IIIa): Y (lla) wherein R 1

R

8

R

14 Y, Z, n and L are as hereinbefore described.

Suitable bases for generating the azomethine ylide of Formula (II) include those derived from alkali metals such as phenyllithium, butyllithium, KNH 2 and NaNH 2 metal carbonates such as potassium carbonate, lithium carbonate, sodium carbonate and cesium carbonate; as well as amines. In preference, the base used is a mono-, di- or tri- .WO 98/50365 PCT/AU98/00312 -7substituted amine, more preferably an alkylamine. Most preferably the base is triethylamine or diisopropylethylamine.

Cyclization of the azomethine ylide may be effected by any suitable means, such as thermal treatment or treatment with metal salts, preferably Cu(I) salts such as Cul.

Preferably cyclization is effected by thermal treatment, such as by heating in optionally boiling solvent. Suitable solvents include tetrahydrofuran, chloroform and 1,2dichloroethane.

In a further preferred aspect, the cyclization of a compound of Formula is followed by oxidative treatment. Oxidative treatment may be performed by means known to and routinely carried out by those skilled in the art. Particularly suitable means include direct oxidation in air, optionally in the presence of silica gel; treatment with Fremy's salt; treatment with quinones such as chloranil or 2,3-dichloro-5,6-dicyano-l,4-benzoquinone (DDQ), and treatment with metal catalysts such as platinum, palladium and nickel.

Preferably the oxidative treatment is effected by DDQ or silica gel in air or Fremy's salt.

When L e of compounds of Formula (III) is iodide, oxidation is promoted.

In a preferred embodiment, a compound of general Formula is prepared by treating a compound of general Formula (III) with triethylamine or diisopropylamine followed by thermally induced cyclization and subsequent oxidative treatment with DDQ or silica gel in air. In a more preferred embodiment a compound of general Formula (la) is prepared by treating a compound of general Formula (liIa) with triethylamine or diisopropylamine followed by thermally induced cyclization and subsequent oxidative treatment with DDQ or silica gel in air or Fremy's salt.

When n is 1, Z is preferably selected from one of carbon, nitrogen, oxygen or sulfur.

More preferably Z is nitrogen or oxygen. Most preferably, Z is oxygen. When n is 2 or 3, preferably one of Z is carbon, preferably the remaining Z are oxygen or nitrogen.

Suitable examples where n is 2 or 3 include A-O-CH 2

A-CH

2

A-O-

CH

2 and A-CH 2

-O-CH

2 -8- Preferably, when W and X, together with the nitrogen and carbon atoms to which they are attached, form an optionally substituted isoquinolinyl or optionally substituted dihydroisoquinolinyl group of general Formula R(i) R r i S wherein R R 4 and R 14 are as defined above.

o Preferably R 1

R

4 are hydrogen, hydroxy, optionally substituted alkyl, optionally substituted alkyloxy, acyloxy, or sulfate. Most preferably they are hydrogen, hydroxy, methoxy, isopropoxy methyl, acetoxy or sulfate. Preferably R' is hydrogen or hydroxy.

When A is an aryl group or an aromatic heterocyclic group, ring A may be an optionally substituted benzene or naphthalene ring or an optionally substituted aromatic heterocyclic group such as pyridine, furan, pyrrole or thiophene and benzene-fused analogues thereof, for example, quinoline, indole, benzofuran and benzothiophene. Attachment of the bicyclic heterocyclic group may be via the benzene or heterocyclic ring. Preferably A is an optionally substituted benzene. Preferably the substituents are hydrogen, hydroxy, optionally substituted alkyl, optionally substituted alkyloxy, acyloxy, or sulfate. Most preferably they are hydrogen, hydroxy, methoxy, iso-propoxy, methyl, acetoxy or sulfate.

WO 98/50365 PCT/AU98/00312 -9- When A is a non-cyclic group RA1RAC-CRA3RA 4

R

A 1

RA

4 are preferably independently selected from hydrogen, optionally substituted alkyl, optionally protected hydroxy, optionally substituted alkoxy or acyloxy. Preferably at least one of RAI A 4 is hydrogen. More preferably at least two are hydrogen. More preferably at least three are hydrogen. Most preferably all R A 1

RA

4 are hydrogen. When R A2 and R A 3 together form a bond so as to form the group RAIC=CRA 4 preferably at least one, or preferably both, of R A 1 and RA 4 are hydrogen.

When A is a cyclic group RARA2C-CRA 3

RA

4 as defined above, preferably R A 2

RA

3 form a 3 to 8-membered cyclic group, preferably 5 to 6-membered. Preferably, R A and

R

3 together with the carbons to which they are attached form a cyclopentane, cyclohexane, cyclopentene, cyclohexene,cyclopentadiene, cyclohexadiene, tetrahydrofuran, dihydrofuran, pyrrolidine, pyrroline, pyran, dihydrophyran, tetrahydropyran or piperidene group. In another preferred form, RA' and RA 4 are hydrogen.

Preferably Y is an optionally substituted phenyl group of Formula (ii):

R"

R

10 R12 (ii) R R 13 Wherein R 9

R

13 are as defined for R 1

R

8 and R 1 4 as described above.

More preferably, R 9

R

13 are hydrogen, hydroxy, optionally substituted alkyl, optionally substituted alkoxy or acyloxy. Most preferably, R 9

R

13 hydrogen, hydroxy, methoxy, iso-propoxy, methyl, acetoxy or sulphate.

The method of the present invention is particularly suitable for the preparation of compounds 1 to 39 as depicted in Tables 1 and 2.

WO 98/50365 WO 9850365PCT/AU98/00312 As used herein the term "alkyl", denotes straight chain, branched or cyclic fully saturated hydrocarbon residues. Unless the number of carbon atoms is specified the term preferably refers to CI- 20 alkyl or cycloalkyl. Examples of straight chain and branched ailkyl include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, amyl, isoamyl, sec-amyl, 1,2-dimethylpropyl, 1, 1-dimethyl-propyl, hexyl, 4-methylpentyl, 1methylpentyl, 2-methylpentyl, 3-methylpentyl, 1,1 -dimethylbutyl, 2 ,2-dimethylbutyl, 3,3dimethylbutyl, 1 ,2-dinethylbutyl, 1 ,3-dimethylbutyl, 1 ,2,2,-trimethylpropyl, 1,1,2trimethyipropyl, heptyl, 5-methoxyhexyl, 1 -methylhexyl, 2 ,2-dimethylpentyl, 3,3dimethylpentyl, 4,4-dimethylpentyl, 1 ,2-dimethylpentyl, 1 ,3-dimethylpentyl, 1,4dimethyl-pentyl, 1,2,3, -trimethylbutyl, 1,1 ,2-trimethylbutyl, 1,1, 3-trimethylbutyl, octyl, 6-methyiheptyl, 1-methylheptyl, 1,1 ,3,3-tetramethylbutyl, nonyl, 6- or 7-methyl-octyl, 4- or 5-ethylheptyl, 2- or 3-propyihexyl, decyl, 3-, 7- and 8-methylnonyl, 5- or 6-ethyloctyl, 3- or 4propylheptyl, undecyl, 8- or 9-methyldecyl, 6or 7-ethylnonyl, 4- or 5-propylocytl, 2- or 3-butyiheptyl, 1-pentylbexyl, dodecyl, 9- or 10-methylundecyl, 7- or 8-ethyldecyl, 5- or 6-propylnonyl, 3- or 4-butyloctyl, 1-2pentylheptyl and the like. Examples of cyclic alkyl include mono- or polycyclic alkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl and the like.

As used herein the term "alkenyl" denotes groups formed from straight chain, branched or cyclic hydrocarbon residues containing at least one carbon-carbon double bond including ethylenically mono-, di- or poly-unsaturated alkyl or cycloalkyl groups as previously defined. Unless the number of carbon atoms is specified the term preferably refers to C 1 2 0 alkenyl. Examples of ailcenyl include vinyl, allyl, I1-methylvinyl, butenyl, isobutenyl, 3-methyl-2-butenyl, 1 -pentenyl, cyclopentenyl, 1 -methyl-cyclopentenyl, Ihexenyl, 3-hexenyl, cyclohexenyl, 1-heptenyl, 3-heptenyl, 1-octenyl, cyclooctenyl, 1nonenyl, 2-nonenyl, 3-nonenyl, 1 -decenyl, 3-decenyl, 1, 3-butadienyl, 1-4 ,pentadienyl, 1 ,3-cyclopentadienyl, 1 ,3-hexadienyl, 1 ,4-hexadienyl, 1 ,3-cyclohexadienyl, 1,4- WO 98/50365 PCT/AU98/00312 11 cyclohexadienyl, 1,3-cycloheptadienyl, 1,3,5-cycloheptatrienyl and 1,3,5,7cyclooctatetraenyl.

As used herein the term "alkynyl" denotes groups formed from straight chain, branched or cyclic hydrocarbon residues containing at least one carbon-carbon triple bond including ethynically mono-, di- or poly- unsaturated alkyl or cycloalkyl groups as previously defined. Unless the number of carbon atoms is specified the term preferably refers to C 1 alkynyl. Examples include ethynyl, 1-propynyl, 2-propynyl, and butynyl isomers, and pentynyl isomers.

The terms "alkoxy, "alkenoxy and "alkynoxy respectively denote alkyl, alkenyl and alkynyl groups as hereinbefore defined when linked by oxygen.

The term "halogen" denotes fluorine, chlorine, bromine or iodine.

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

The term "heterocyclic" denotes mono- or polycarbocyclic groups wherein at least one carbon atom is replaced by a heteroatom, preferably selected from nitrogen, sulphur and oxygen. Suitable heterocyclic groups include N-containing heterocyclic groups, such as, unsaturated 3 to 6 membered heteromonocyclic groups containing 1 to 4 nitrogen atoms, for example, pyrrolyl, pyrrolinyl, imidazolyl, imidazolinyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl or tetrazolyl; saturated 3 to 6-membered heteromonocyclic groups containing 1 to 4 nitrogen atoms, such as, pyrrolidinyl, imidazolidinyl, piperidyl, pyrazolidinyl or piperazinyl; condensed saturated or unsaturated heterocyclic groups containing 1 to 5 nitrogen atoms, WO 98/50365 PCT/AU98/00312 12such as, indolyl, isoindolyl, indolinyl, isoindolinyl, indolizinyl, isoindolizinyl, benzimidazolyl, quinolyl, isoquinolyl, indazolyl, benzotriazolyl, purinyl, quinazolinyl, quinoxalinyl, phenanthradinyl, phenathrolinyl, phthalazinyl, naphthyridinyl, cinnolinyl, pteridinyl, perimidinyl or tetrazolopyridazinyl; saturated 3 to 6-membered heteromonocyclic groups containing 1 to 3 oxygen atoms, such as tetrahydrofuranyl, tetrahydropyranyl, tetrahydrodioxinyl, unsaturated 3 to 6-membered hetermonocyclic group containing an oxygen atom, such as, pyranyl, dioxinyl or furyl; condensed saturated or unsaturated heterocyclic groups containing 1 to 3 oxygen atoms, such as benzofuranyl, chromenyl or xanthenyl; unsaturated 3 to 6-membered heteromonocyclic group containing 1 to 2 sulphur atoms, such as, thienyl or dithiolyl; unsaturated 3 to 6-membered heteromonocyclic group containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, such as, oxazolyl, oxazolinyl, isoxazolyl, furazanyl or oxadiazolyl; saturated 3 to 6-membered heteromonocyclic group containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, such as, morpholinyl; unsaturated condensed heterocyclic group containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, such as, benzoxazolyl or benzoxadiazolyl; unsaturated 3 to 6-membered heteromonocyclic group containing 1 to 2 sulphur atoms and 1 to 3 nitrogen atoms, such as, thiazolyl, thiazolinyl or thiadiazoyl; saturated 3 to 6-membered heteromonocyclic group containing 1 to 2 sulphur atoms and 1 to 3 nitrogen atoms, such as, thiazolidinyl; and unsaturated condensed heterocyclic group containing 1 to 2 sulphur atoms and 1 to 3 nitrogen atoms, such as, benzothiazolyl or benzothiadiazolyl.

The term "acyl" refers to a carboxylic acid residue wherein the OH is replaced with a residue, for example, as defined for W, X, and Y and specifically may denote carbamoyl, aliphatic acyl group or acyl group containing an aromatic ring, which is referred to as aromatic acyl or a heterocyclic ring, which is referred to as heterocyclic acyl, preferably

C

1 2 0 acyl. Examples of suitable acyl include carbamoyl; straight chain or branched WO 98/50365 WO 9850365PCT/AU98/00312 13 alkanoyl such as formyl, acetyl, propanoyl, butanoyl, 2-methyipropanoyl, pentanoyl, 2,2dimethyipropanoyl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl, undecanoyl, dodecanoyl, tridecanoyl, tetradecanoyl, pentadecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl and icosanoyl; alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl, t-pentyloxycarbonyl and heptyloxycarbonyl; cycloalkylcarbonyl such as cyclopropylcarbonyl cyclobutylcarbonyl, cyclopentylcarbonyl and cyclohexylcarbonyl; alkylsulfonyl such as methylsulfonyl and ethylsulfonyl; alkoxysulfonyl such as methoxysulfonyl and ethoxysulfonyl; aroyl such as benzoyl, toluoyl and naphthoyl; aralkanoyl such as phenylalkanoyl phenylacetyl, phenyipropanoyl, phenylbutanoyl, phenylisobutylyl, phenylpentanoyl and phenyihexanoyl) and naphthylalkanoyl naphthylacetyl, naphthylpropanoyl and naphthylbutanoyl]; aralkenoyl such as phenylalkenoyl phenyipropenoyl, phenylbutenoyl, phenylmethacryloyl, phenylpentenoyl and phenyihexenoyl and naphthylalkenoyl naphthylpropenoyl, naphthylbutenoyl and naplithylpentenoyl); aralkoxycarbonyl such as phenylalkoxycarbonyl benzyloxycarbonyl); aryloxycarbonyl such as phenoxycarbonyl and napthyloxycarbonyl; aryloxyalkanoyl such as phenoxyacetyl and phenoxypropionyl; arylcarbamoyl such as phenylcarbamoyl; aryithiocarbamoyl such as phenyithiocarbainoyl; aryiglyoxyloyl such as phenyiglyoxyloyl and naphthylglyoxyloyl; arylsulfonyl such as phenylsulfonyl and napthylsulfonyl; heterocycliccarbonyl; heterocyclicalkanoyl such as thienylacetyl, thienyipropanoyl, thienylbutanoyl, thienylpentanoyl, thienyihexanoyl, thiazolylacetyl, thiadiazolylacetyl andtetrazolylacetyl; heterocyclicalkenoyl such as heterocyclicpropenoyl, heterocyclichutenoyl, heterocyclicpentenoyl and heterocyclichexenoyt; and heterocyclicglyoxyloyl such as thiazolyiglyoxyloyl. and thienyiglyoxyloyl.

The term "acyloxy" refers to acyl, as herein before defined, when linked by oxygen.

In this specification "optionally substituted" is taken to mean that a group may or may not be further substituted or fused (so as to form a condensed polycyclic group) with one or more groups selected from alkyl, alkenyl, alkynyl, aryl, halo, haloalkyl, haloalkenyl, WO 98/50365 PCT/AU98/00312 -14haloalkynyl, haloaryl, hydroxy, alkoxy, alkenyloxy, aryloxy, benzyloxy, haloalkoxy, haloalkenyloxy, haloaryloxy, nitro, nitroalkyl, nitroalkenyl, nitroalkynyl, nitroaryl, nitroheterocyclyl, amino, alkylamino, dialkylamino, alkenylamino, alkynylamino, arylamino, diarylamino, benzylamino, dibenzylamino, acyl, alkenylacyl, alkynylacyl, arylacyl, acylamino, diacylamino, acyloxy, alkylsulphonyloxy, arylsulphenyloxy, heterocyclyl, heterocycloxy, heterocyclamino, haloheterocyclyl, alkylsulphenyl, arylsulphenyl, carboalkoxy, carboaryloxy mercapto, alkylthio, benzylthio, acylthio, cyano, nitro sulfate and phosphate groups.

As used herein, the term "protecting group", refers to an introduced functionality which temporarily renders a particular functional group inactive. The term "protected hydroxy" refers to a hydroxy group which has been temporarily rendered inactive by a protecting group. 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).

As used herein, "heteroatom" refers to any atom other than a carbon atom which may be a member of a cyclic organic compound. Examples of suitable heteroatoms include nitrogen, oxygen, sulfur, phosphorous, boron, silicon, arsenic, sellenium and telluruim.

As used herein, the term base" refers to any proton acceptor/electron pair donator suitable for the generation of an azomethine ylide.

The term "synthon" is taken to refer to a structural or chemical equivalent for a desired functional unit and which can be converted to the desired unit by known or conceivable synthetic operations 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 stabilize the negative charge which it carries such as the halogens, sulfates as hereinbefore defined, 15 protonated alcohols and ethers, pyridinium salts; iminium salts such as derived from ,diclohexylcarbodiimide (DCC) and diazonium ions.

The present invention is hereinafter described using a compound of Formula (II) prepared bythe hereinafter describedi iethods. This is done, however, with the understanding that the present invention extends to compounds of Formula (II) prepared by any other Smeans Accordingly, another aspect of the invention relates to a process for the preparation of a 10 compound of Formula comprising: a) coupling a compound of Formula (IV) with a compound of Formula Hal A -P4 (V

(IV)

to afford the compound of Formula (VI): 20

A

(VI)

PZn wherein PZn is a synthon for Zn; b) unmasking Zn of compound (VI) and coupling with a compound L-CH 2 to provide compound (VII):

A

O (vi) -o0

L-

16wherein L and L' are each a leaving group or a substituent convertible to a leaving group.

c) treatment of compound (VII) with animine of Formula (VIII)

(VI

d) generation of the azomethine ylide of general formula (II) and subsequent cyclization of the ylide: wherein Hal is a halogen and A, W, X, Y, Z and n are as hereinbefore described.

Ina preferred embodiment, the present invention relates to a process for the preparation qfta compound of Formula (la) comprising: coupling a compound of Formula (IV) with a compound of Formula (Va):

R

.y Hal R7 PZn Re (IV) (va) to afford the compound of Formula (VIa):

R

5

R

6 S M(VIa)

PIRS

(Via) unmasking Zn of compound (Via) and coupling with a compound L-CH,-C(O)-L' to provide compound (VIIa): WO 98/50365 PCT/AU98/00312 -17- Rs Re SR(VIIa) Re 0 L-

O

treatment of compound (VIIa) with a compound of Formula (VIIIa): R4 103 s (VIIIa) R2 R14

R

1 generation of the azomethine ylide of general Formula (IIa) and subsequent cyclization of the ylide; wherein Hal, L, PZ Y, Z, R'-R 8

R

14 and n are as hereinbefore defined.

Preferably Y is optionally substituted phenyl of Formula More preferably, Y is phenyl substituted with optionally substituted alkyl, optionally substituted alkoxy or acyloxy. Most preferably, Y is phenyl substituted with hydrogen, hydroxy, methoxy, methyl or acetoxy.

In a preferred aspect, P is a protecting group for Z n and unmasking Z n refers to removal of the protecting group. Removal of the protecting group P may be carried out under routine conditions known to those skilled in the art, for example as described in Protective Groups in Organic Synthesis. Preferably, P is a protecting group which is labile under hydrolysis conditions. Even more preferably, P is acetyl. In a preferred embodiment, n is 1, Z is oxygen and P is acetyl.

.WO 98/50365 PCT/AU98/00312 -18- In another preferred aspect, wherein the terminal Z is oxygen, PZ n is an aldehyde or acyl group. Unmasking of Z n comprises oxidation, such as Baeyer-Villiger oxidation, of the aldehyde or acyl to a corresponding ester followed by hydrolysis.

Other suitable synthons are known to those skilled in the art.

Preferred L' is halogen, most preferably chlorine or bromine, or OH converted to a leaving group preferably by reaction with DCC.

The coupling of compounds of general Formula (IV) with those of Formula may be suitably carried out under conditions known and routinely employed by those skilled in the art, for example in the presence of catalysts such as Pd(PPh 3 4 PdCl2 (PPh 3 2 or Cu(I) mediated conditions, CuI, and such as those described by Sonogashira in Comprehensive Organic Synthesis, (Ed. B.M. Trost and I. Fleming, Peramon Press, New York, 1991, Vol. 3, 521).

Schemes 1 to 4 provide a schematic overview of representative methods of the invention.

Scheme Ia OQAc (bXi) -AcO HO AcO HO "N I o (bXii) Br Br (and bond-shift isomers) a Key: Phenylacetylene 1.1 equiv., Pd(PPh 3 4 0.01%, Cul 0.02%, Et 3 N, 18 0 C, 4h. K 2

CO

3 1.5 equiv., MeOH, 0.25 h; (ii) 2-bromoacetic acid, 1 equiv., DCC 1.05 equiv., DMAP 0.05 equiv., 18 0 C, (i) Isoquinoline 1 equiv., THF, 18°C, 36h then Et 3 N 1 equiv., CHCI 3 reflux, 8h; DDQ 1 equiv., CH 2

C

2 18C, 2h WO 98/50365 WO 9850365PCT/AU98/0031 2 19- Scheme 2 a ~0

K-OH

OMe (a) (d) (a) (c) OMe Pr'O =OPr MeO c (fM OMe Pr O /OPrj MP HO Me jg Pr'O Prio MeO OPr' MeD 1 MeO0 MeO"0 pr' HO MeO OH MeO MeO 0 MeO"0

HO

lamnellarin K a Key: K 2 C0 3 2 equiv., Pr'Br 1.3 equiv., DM F, 80 0 C, 13h 12 1.1 equiv., AgO 2

CCF

3 1.1 equiv., 65 0

C,

7h CBr 4 2 equiv., PPh 3 2 equiv., Zn 2 equiv., nBuLi 2 equiv. added to solution of the gemndibromostyrene in THE at -781C then ZnCI 2 1.1 equiv. -78 1800 then aryliodide 1 equiv., Pd(PPh 3 4 0.02 equiv., 1800 1.5h mnCPBA 1.3 equiv., KHCO 3 3 equiv., CH 2

CI

2 0 18'C 2-lodoacetic acid 1.1 equiv., DCC 1.1 equiv., DMAP 0.05 equiv., CH 2

CI

2 1800, 4h 3,4-Dihydro-6,7-dimethoxy- 1. 1 equiv., 1,2-dichioroethane (solvent), 1800C, 8h then Pe 2 NEt 1lequiv., 83 0 C, 32h (81%) AIC1 3 3.6 equiv., CH 2

CI

2 180C, 4h WO 98/50365 WO 9850365PCT/AU98/00312 20 Scheme 3 a

I

(b) MeO Pr 1 O 0- OMe MeO- /OPr Ps 0 .4 (d) OMe Me0G \5 /Or P5 0 HO MeO N MeO) (e OMe MeO MeO OH Mf HO MeO MeO N MeG lamnellarin T Iamnellarin T diacetate 8 Key: CBr 4 2 equiv., PPh 3 2 equiv., Zn 2 equiv., nBuLi 2 equiv. added to solution of the gemdbromostyrene in THF at -78 0 C then ZnC1 2 1.1 equiv. -78 180C then aryl iodide 1 equiv., PdCI 2 (PPh 3 4 0.005 equiv., 180C 1.5h mnCPBA 1.2 equiv., KHCO 3 3 equiv., CH 2 CI, 0 2-lodoacetic acid 1.05 equiv., DCC 1.05 equiv., DMAP 0.01 equiv., C- 2

CI

2 1800-, 4h 3,4-Dihydro-5,6,7trimelhoxyisoquinoline 1.2 equiv., 1,2-dichioroethane (solvent), 18 0 C, 15h then Pr 1 2 NEt lequiv., 2400, 32h AIC1 3 2.4 eqluiv., CH 2

CI

2 ,1800, 0. h Ac 2 O/pyridine 1, solvent), DMAP cat., 1 81C, 26h WO 98/50365 WO 9850365PCT/AU98/00312 -21- Scheme 4 8 MeO)C (a) MeO Meo 0 Meo 0O Me O H MeO\ MeON 0 I -deoxylamnellarin K MeO (eoc Meo Meo OPr Mao 0 d MaO"0 lamellarin

U

rMeo Me OPr o Me

MOO

Meo Meo OH HO Me 1 0 Mao Mao MeO lamnelianin W a Key: 3,4-Dihyd r-6,7-dimethaxyisoq Lino line 1.2 equiv., 1,2-dichloroethane (solvent), 1 BIC, 15h then Pr' 2 NEt 1.05 equiv., 8300, 28h AIC1 3 3equiv., CH 2

CI

2 18 0 C, 16h (c) 3,4-D ihydro-6,7-d imethoxyisoqtuno line 1.2 equiv., 1,2-dichloroethane (solvent), 18 0 C, 17h then Pr' 2 N Et 1.05 equiv., 8300, 20h Wd)AJC1 3 3.3 equjv., CH 2

CI

2 18-C, 14h DDQ 1.25 equiv., 0HC1 3 650C, 211 (f) AICd 3 3.3 equiv., CH 2

CI

2 18D0, 17h WO 98/50365 PCT/AU98/00312 -22- The method of the present invention encompasses the synthesis of a large group of compounds. Traditionally, drug candidates are 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 effectuating the generation of large libraries of molecules and has been successfully exploited in the synthesis and evaluation of small organic molecule libraries. These libraries may exist as molecules in free solution or linked to a solid phase, for example, polymer beads, pins, microtitre plates or microchips. 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.

Solution phase libraries may be prepared via parallel syntheses wherein different compounds are synthesised in separate reaction vessels in parallel, often in an automated 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 support containing the compounds prepared in the prior step can be split into a number of batches, treated with the appropriate reagent and recombined. By performing one or more of the steps a) as hereinbefore described, in a parallel or split fashion, in solution phase or on solid support, the present invention is amenable to the generation of large numbers of compounds of general Formula Accordingly, another aspect of the present invention provides a means for generating compounds of Formula by performing one or more of the following steps: the coupling of a compound of Formula (IV) with a compound of Formula unmasking Zn of the compound of Formula (VI) and coupling with a compound L-

CH

2 treatment of the compound of Formula (VII) with the imine of Formula (VIII), in a parallel or split fashion, in solution phase or on solid support.

-23 A, further aspectof the 'invention contemplates compounds .of thegeraFoml(I) Y

A

0fi x w Yet another aspect of the invention contemplates",a compound of general Formula (III): Zn(I) .4..01I) -0 x Le w WO 98/50365 PCT/AU98/00312 -24- Another aspect of the invention contemplates the compounds of the general Formula (I) when prepared by the methods herein described.

Yet another aspect of the present invention contemplates a method of treatment comprising the administration of a treatment effective amount of a compound of general Formula as an active ingredient, to an animal, including a human, in need thereof.

Preferably the compound of general Formula is prepared by the methods as hereinbefore described.

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 10 g per kg of body weight per dosage. Preferably, the dosage is in the range of 1 /g to 10 g per kg of body weight per dosage. More preferably, the dosage is in the range of 1 mg to 10 g per kg of body weight per dosage. Even more preferably, the dosage is in the range of 1 mg to 5 g per kg of body weight per dosage. More preferably, the dosage is in the range of 1 mg to 2 g per kg of body weight per dosage. More preferably, the dosage is in the range of 1 mg to 1 g per kg of body weight per dosage.

In a preferred embodiment, the method of treatment relates to treating multidrug resistant tumors.

In another embodiment, the method of treatment contemplates improving the antitumor chemotherapeutic effect of multidrug resistant affected drugs.

In another preferred embodiment, the method of treatment is a method for inducing apoptosis. More preferably, the method of treatment is a method of inducing apoptosis on a multidrug resistant cell WO 98/50365 PCT/AU98/00312 In another embodiment, the method of treatment contemplates modulating immunological functions.

The active ingredient may be administered in a single dose or a series of doses. While it is possible for the active ingredient to be administered alone, it is preferable to present it as a composition, preferably as a pharmaceutical composition.

Yet another aspect of the invention contemplates compositions comprising a compound of general Formula together with a pharmaceutically acceptable carrier, excipient or diluent. Preferably the compound of general Formula is prepared by the methods as hereinbefore described.

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.

WO 98/50365 PCT/AU98/00312 -26- 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 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 starch glycolate, cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose) surface-active or dispersing agent. Moulded tablets may be made by moulding 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 provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release 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, or sucrose and acacia gum; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

Compositions for rectal administration may be presented as a suppository with a suitable base comprising, for example, cocoa butter.

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.

Compositions suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions which may contain anti-oxidants, buffers, bactericides and solutes which render the composition isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending WO 98/50365 PCT/AU98/00312 -27agents and thickening agents. The compositions may be presented in unit-dose or multidose sealed containers, for example, ampoules and vials, and may be stored in a freezedried (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 subdose, 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 methacrylie 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.

The present invention also provides the use of a compound of general Formula for the manufacture of a medicament for treatment of an animal or human in need thereof.

Preferably the compound of general Formula is prepared by the methods as WO 98/50365 PCT/AU98/00312 28 hereinbefore described.

Another aspect of the invention contemplates an agent comprising a compound of general Formula for the treatment of an animal or human in need thereof. Preferably the compound of general Formula is prepared by the methods as hereinbefore described.

In a first embodiment, the agent is for treating multidrug resistant tumors.

In another embodiment the agent is for inducing apoptosis on a multi-drug resistant cell.

In yet another embodiment, the agent is for improving the anti-tumour chemotherapeutic effect of multidrug resistant affected drugs.

A further embodiment is an agent for modulating immunological functions.

Yet another aspect of the invention contemplates the use of a compound of general Formula for the treatment of an animal or human in need thereof. Preferably the compound of general Formula is prepared by the methods as hereinbefore described.

In a preferred embodiment, the use is in the treatment of multidrug resistant tumours.

In a further embodiment, the use is in improving the chemotherapeutic effect of multidrug resistant affected drugs.

Yet another embodiment is the use in modulating immunological functions.

In another embodiment, the invention contemplates the use of a compound of general Formula for inducing apoptosis in an animal or human in need thereof. Preferably apoptosis in a multidrug resistant cell.

WO 98/50365 PCT/AU98/00312 -29- The following abbreviations as used in the hereinafter described embodiments of the present invention.

DCC Dicyclohexylcarbodiimide DDQ 2,3-Dichloro-5,6-dycano- 1,4-benzoquinone DMAP 4-(N,N)-dimethylaminopyridine THF Tetrahydrofuran In one embodiment of the invention, Compound 1 was prepared by Sonogashira crosscoupling of phenylacetylene and o-iodophenylacetate to give, after hydrolysis of the initial coupling product, o-hydroxytolan. This last compound was reacted with bromoacetylbromide under standard conditions to deliver the ester which was then treated with isoquinoline to give the isoquinoline salt which was immediately treated with triethylamine (so as to generate the associated azomethine ylide) in refluxing THF and the mixture of dihydropyrrole-type cycloaddition products thereby obtained were subjected to oxidation with either 2 3 -dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) or silica gel in air. In this manner, the target Compound 1 was obtained and its structure established by single-crystal X-ray analysis.

In another embodiment, subjection of a more highly oxygenated tolan-ester to reaction with 6 7 -dimethoxy-3,4-dihydroisoquinoline under the same conditions as employed in the formation of Compound 1 provided the 8,9-dihydro-congener Compound 34.

Deprotection of Compound 34 with BCl 3 or AIC13 then gave Compound The present invention is now described with reference to the following non-limiting Examples.

WO 98/50365 PCT/AU98/00312 Example 1.

4-Phenyl-6H-[1]Benzopyrano[4',3':4,5]pyrrolo[2,1-a]isoquinolin-6-one (Compound 1) o-Acetoxytolan: Pd(PPh3)4 (132 mg 0.114 mmol), phenylacetylene (1.3 mL, 1.20 g, 12.6 mmol) and Cul (44 mg, 0.23 mmol) were added, sequentially, to a solution of 2acetoxyiodobenzene (3.0 g, 11.45 mmol) in Et3N (20 mL) and the reaction mixture stirred for 4 h at room temperature. The reaction mixture was then concentrated under vacuum at room temperature. The residue was dissolved in CH2C12 (50 mL) and the resulting solution washed with HCI (1 x 50 ml of a 0.5 M aqueous solution) and brine (1 x 50 ml) then dried (MgSO4), filtered and concentrated on to silica gel (8 This solid was subjected to chromatography (silica gel; 3:1, 2:1, 1:1 then 1:2 hexane/CH2Cl2 elution) and concentration of the appropriate fractions then gave pure o-acetoxytolan (2.7 g, 100 Spectra of this compound are identical with those previously reported Arcadi,S. Cacchi, M. D. Rasario, G. Fabrizi and F.

Marinelli, J. Org. Chem., 1996, 61, 9280).

o-(a-Bromoacetoxy)tolan o-Acetoxytolan (2.24 g, 9.50 mmol) was added to a stirring slurry of K2C03 (2.0 g, mmol) in methanol (20 mL). After 15 min. the reaction mixture was diluted with HC1 (50 ml of a 0.5 M aqueous solution) and extracted with CH2C12 (2 x 40 mL). The combined extracts were dried (MgSO4) and then concentrated under reduced pressure. The solid residue [pure o-hydroxytolan by 1 H nmr] obtained in this manner was dissolved in CH2C12 (20 mL) and a-bromoacetic acid (1.32 g, 9.5 mmol) and DMAP (58 mg, 0.48 mmol) added. Dicyclohexyl carbodiimide (DCC) (2.07 g, 10 mmol) was then added portionwise and the reaction mixture stirred at room temperature for 3 h, then filtered through Celitea and the filtrate concentrated on to silica gel (8 This solid was subjected to flash chromatography on silica gel (sequential elution with 2:1, 1:1 then 1:2 hexane/CH2Cl2), concentration of the appropriate fractions then gave the title compound 2.73 g, 91.2%) as an amber coloured oil.

IR(KBr neat, cm-1) 3060, 2221, 1761, 1596, 1571, 1496, 1444, 1258, 1195, 1116, 1099. 1H NMR (300 MHz, CDCl 3 5 7.61 (dd, J 1.5, 7.5 Hz, 1H), 7.54-7.52 2H), 7.41-7.35 (m, 4H), 7.29 (dd, J 1.5, 7.5 Hz, 1H), 7.17 (dd, J 1.4, 8.1Hz, 1H), 4.13 2H). 13 C NMR APT (75.5 MHz, CDCl 3 5 165.0 150.5 133.0 131.4 129.3 (CH), .WO 98/50365 PCT/AU98/00312 -31 128.2 126.3 122.4 121.7 116.9 94.6 83.5 25.1 (CH2). MS eV) m/z 316 (31) 314 (30) 193 (100, M COCH2Br). Anal. Calcd for

C

16

HINO

2 Br: C, 60.98; H, 3.52; Br, 25.35. Found: C, 61.26; H, 3.45; Br, 25.06.

14-Phenyl-6H-[1]benzopyrano[4', 3 ':4,5]pyrrolo[2, 1-a]isoqinolin-6-one: Isoquinoline (98 pl, 0.83 mmol) was added to a solution ofo-(ao-bromoacetoxy)tolan (262 mg, 0.83 mmol) in THF mL) and the resulting mixture allowed to stir at room temperature for 6 h. Addition of Et3N (215 tl, 0.83 mmol) and chloroform (10 mL) then gave a bright orange solution which was heated at reflux for 4 h at which point a light-yellow solution was achieved. The cooled reaction mixture was concentrated under reduced pressure, the residue dissolved in CH2C12 (7 ml) and DDQ (189 mg, 0.83 mmol) added. The reaction mixture was immediately concentrated on to silica gel (2 g) and subjected to flash chromatography on silica gel (sequential elution with 2:1, 1:1 then 1:2 hexane/CH2Cl2). Concentration of the appropriate fractions then gave the title compound (277 mg, 92.1%) as white crystalline masses, mp 313- 5"C.IR(KBrdisc, cm- 1 3047, 1705, 1536, 1470, 1445, 1411, 1369, 1312, 1179, 1110, 1049.

1H NMR (300 MHz, CDCI 3 6 9.29 J 7.2Hz, 1H), 7.67 J 7.2 Hz, 1H), 7.64-7.62 (m, 2H), 7.56-7.39 4H), 7.32 (dt, J= 1.8, 7.5 Hz, 1H), 7.25 (dt, J= 1.5, 7.5 Hz, 1H), 7.12-7.05 2H), 6.99 (dt, J 1.5, 7.5 Hz, 1H). 13 C NMR APT (75.5 MHz, CDCI 3 6 155.2 151.7 135.6 134.1 131.0 129.9 128.7 128.6 128.4 (CH), 128.1 127.5 127.3 125.0 124.4 124.0 123.9 117.9 117.3 114.3 113.4 109.3 MS (70 eV) m/z 361 (100, M); HRMS calcd for C 25

H

15

NO

2 361.1103. Found 361.11031. X-ray obtained.

Example 2.

Lamellarin K triisopropylether (Compound 36) Isovanillin isopropyl ether: Isopropyl bromide (19.0 mL, 200 mmol) was added to a suspension of K2CO3 (42.0 g, 302 mmol) and isovanillin (23 g, 151.3 mmol) in DMF (100 gmL) and the stirring slurry heated to 80"C for 13 h. The reaction mixture was then cooled WO 98/50365 PCT/AU98/00312 -32to room temperature, diluted with ether (200 mL) and washed with H20 (4 x 200 mL), dried (MgSO4) and concentrated under reduced pressure giving the title compound as a slightly tan oil (29.3 g, 100%) which required no further purification. The spectra of this material are identical with those previously reported Ishii, Chen and T. Ishikawa J. Chem. Soc., Perkin Trans. 1, 1987, 671.).

Vanillin isopropyl ether: Isopropyl bromide (40.0 mL, 421 mmol) was added to a suspension of K2C03 (48.0 g, 348 mmol) and vanillin (40 g, 263 mmol) in DMF (150 mL) and the stirring slurry heated to 80"C for 15 h. The reaction mixture was then cooled to room temperature, diluted with diethyl ether (200 mL) and washed with H20 (4 x 200 mL), dried (MgSO4) and concentrated under reduced pressure giving the title compound as a slightly tan oil (51.0 g, 100%) which required no further purification. The spectra of this material are identical with that previously reported F. Comber and M. V. Sargent, J. Chem. Soc., Perkin Trans. 1, 1991, 2783).

2 -Iodo-5-isopropoxy-4-methoxybenzaldehyde: Silver trifluoroacetate (12.3 g, 56.7 mmol) was added to a solution of isovanillin isopropyl ether (10.0 g, 51.5 mmol) in dry chloroform (120 mL) under nitrogen and the resultant slurry stirred and heated to 61"C. Iodine (14.4 g, 56.7 mmol) was then added portionwise (6 portions) over 0.6 h and the reaction allowed to reflux for a futher 6.5 h. The reaction mixture was then cooled and filtered, rinsing with chloroform mL). The filtrate was washed with Na2S205 (10 solution in water, 150 mL), NaHCO3 (sat. in water, 150 mL) and water (150 mL) dried (MgSO4) and concentrated under vacuum.

The solid residue was suspended in hexane (200 mL) and stirred vigoursly for 1 h, cooled in an ice bath for 1 h and filtered rinsing with ice cold hexane (200 mL) to give the title compound as a cream solid (15.3 g, 93%) mp 75-6"C. IR (KBr disc, cm- 1 3072, 2978, 2932, 1673, 1581, 1503, 1438, 1385, 1332, 1260, 1215, 1174, 1157, 1133, 1105, 1024, 939. 1 H NMR (300 MHz, CDC13) 8 9.77 7.34 1H), 7.24 1H), 4.57 (septet, J= 6.0 Hz, 1H), 3.86 3H), 1.31 J= 6.0 Hz, 6H). 13 C NMR APT (75.5 MHz, CDC 3 6 194.7 155.5 147.9 128.2 122.2 114.1 110.8 92.4 71.3 56.4 21.77 MS WO 98/50365 PCT/AU98/00312 -33eV) m/z 320 (M 43), 278 (100), 249 207 150 Anal. Calcd for CliH13031: C, 41.27; H, 4.09; I, 39.64. Found: C, 41.06; H, 3.80; I, 39.59.

P,t-Dibromo-4-isopropoxy-3-methoxystyrene: Carbon tetrabromide (51.3 g, 154.7 mmol) was added portion-wise to a magnetically stirred mixture of zinc dust (10.1 g, 154.5 mmol) and PPh3 (40.5 g, 159.4 mmol) in CH2C12 (350 ml) maintained at 0 oc on an ice-salt bath. This suspension was allowed to warm to room temperature and then stirred at for a further 22 h. After this time the reaction mixture was re-cooled to 0"C and vanillin isopropyl ether (15.0 g, 77.3 mmol) was added dropwise over 2 min. and allowed to stir at room temperature for 1 h. After dilution with hexane (200 ml) the resulting mixture was filtered through a sintered glass funnel and the filtrate concentrated on to silica gel (30 The resulting solid was added to the top of a flash chromatography column (silica gel: 20 cm long x 10 cm wide) which was subjected to gradient elution 1:1 and then 1:2 hexane/CH 2 Cl 2 Concentration of the appropriate fractions (Rf in 1:1 hexane/CH2Cl2) then afforded the title compound (27.2 g, 99.8%) as white crystalline masses, m.p. 36-38"C. IR (KBr disc, cm- 1 2976, 2973, 1599, 1510. 1464, 1418, 1383, 1373, 1267, 1235, 1141, 1110, 1036. 1 H NMR (300 MHz, CDCl 3 7.41 1H), 7.21 J= 2.1 Hz, 1H), 7.07 (dd, J= 2.1, 8.4 Hz, 1H), 6.87 J= 8.4 Hz), 4.58 (septet, J= 6.0 Hz, 1H), 3.87 (s, 3H), 1.39 J= 6.0 Hz, 6H). 13 C NMR+ APT (75.5 MHz, CDC13) 6 149.7 147.7 136.5 127.9 121.9 114.4 111.8 87.1 71.1 56.0 (CH3), 22.1 (CH3). MS (70 eV) m/z 352 350 448 (31) (M 310 308 (100), 306 (54) CH2CHCH 3 295 293 291 (36) (M CH2CHCH3 CH3). Anal. Calcd for C 1 2

HI

4 0 2 Br 2 41.17; H, 4.03; Br, 45.65. Found: C, 41.21; H, 3.92; Br, 45.47.

2-[(4-Isopropoxy-3-methoxy-phenyl) ethynyl]-5-isopropoxy-4-methoxybenzaldehyde: n- Butyllithium (5.25 mL of a 2.5 M solution in hexane, 13.15 mmol) was added dropwise over 3 mins to a magnetically stirred solution of P,P-dibromo-4-isopropoxy-3-methoxystyrene (2.3 g, 6.58 mmol) in THF (30 mL) maintained at -78 OC on a dry-ice acetone bath. The slightly tan coloured solution thereby obtained was allowed to stir for a further 50 min at -78 OC then anhydrous ZnCl2 (dried under high vacuum at 120 OC for 20 h) was added to the reaction mixture which slowly became colourless as it was warmed to room temperature over 1 h.

.WO 98/50365 PCT/AU98/00312 -34- Pd(PPh3)4 (145 mg, 0.125 mmol) and aldehyde (2.0 g, 6.26 mmol) were added and the reaction mixture stirred at 18"C for 4 h. After this time the reaction mix was diluted with ethyl acetate (150 mL) and washed with brine (2 x 100 ml) then dried (MgSO4), filtered and concentrated on to silica (10 The resulting solid was added to the top of a flash chomatography column column (10 cm long x 5 cm wide) which was subject to gradient elution 1:2, hexane/CH2Cl2 CH2C12, then 9:1 CH2Cl2/ethyl acetate). Concentration of the appropriate fractions (Rf 0.4 in CH2Cl2) then afforded the title compound as white crystalline masses, mp 121-2"C. IR (KBr disc, cm- 1 2978, 2933, 2837, 2204, 1687, 1589, 1515, 1509, 1471, 1397, 1357, 1275, 1241, 1217, 1133, 953. 1H NMR (300 MHz, CDCI 3 6 10.48 1H), 7.41 1H), 7.12 J= 8.1 Hz, 1H), 7.04 2 6.87 J= 8.1 Hz, 1H), 4.68 (septet, J= 6.0 Hz, 1H), 4.58 (septet, J= 6.0 Hz, 1H), 3.95 3H), 3.88 3H), 1.40 (2 x d, 2 x J= 6.0 Hz, 2 x 6H). 13

C

NMR APT (75.5 MHz, CDCI 3 6 190.2 154.4 149.7 148.1 147.6 129.6 124.7 121.3 114.6 114.5 114.3 110.6 94.9 83.4 71.0 70.9 56.0 (CH3), 55.7 (CH3), 21.8 (CH3), 21.6 (CH3). MS (70 eV) m/z 382 340 M* CH2CHCH3), 298 (100, M 2 x CH2CHCH3), 283 M 2 x CH2CHCH3 CH3). Anal. Calcd for C23H2605: C, 72.23; H, 6.85. Found: C, 72.08; H, 6.92.

2 4 -Isopropoxy-3-methoxy-phenyl)ethynyl]-5-isopropoxy-4-methoxyphenol: m- Chloroperoxybenzoic acid [1.2 g, ALDRICH, 50% (remainder 3-chlorobenzoic acid and water), ca. 7.0 mmol] was added in portions over 0.25 h to a magnetically stirred mixture of isopropoxy- 3 -methoxy-phenyl)ethynyl]-5-isopropoxy-4-methoxybenzaldehyde (2.20 g, 5.75 mmol) and KHCO 3 (1.73 g, 17.3 mmol) in CH2C12 (50 mL) maintained at 0"C (ice-bath). The resulting slurry was stirred for a further 1 h between 0"C and 18"C then filtered through Celite® and the solids thus retained rinsed with CH2C12 (1 x 50 mL). The combined filtrates were concentrated under reduced pressure and the residue treated with NH 3 (30 ml of a saturated methanolic solution). After 1 h at 18"C the reaction mixture was concentrated under reduced pressure and the residue redissolved in CH2C12 (200 mL) then concentrated on to silica gel (8 g, 400-200 mesh). The resulting powder was loaded on top of a column of silica (40-10 mesh TLC grade, 10 cm wide x 5 cm long) and eluted with 2:1, 1:1 and 1:2 hexane/CH2Cl2 then neat WO 98/50365 WO 9850365PCT/AU98/00312 35 CH2C12 (250 ml of each). The appropriate fractions (Rf 0.3 in 1:2 hexane/CH2C12) were concentrated under reduced pressure to give the title compound (1.97 g, 92 as white crystalline masses, mp 130-1 LR (KBr disc, cm-i1) 3404, 2981, 293 5, 1620, 1573, 1513, 1467, 1450, 1418, 1383, 1373, 1330, 1269, 1240, 1214, 1166, 1135, 1113, 95 1. IH NMvfp (300 M&,I CDCl 3 )5 7.03 (dd,J= 1.8, 8.4 Hz, 1H), 7.00 J= 2.1 Hz, 1H), 6.87 III), 6.81 J= 8.4 Hz, 1 6.53 1ff), 5.81 111), 4.52 (septet J= 6.0 Hz' 2H), 3.82 3H), 3.78 3H), 1.35 J= 6.0 Hz, 12H). 13 C NMiR+ APT (75.5 MIHz, CDCI 3 5 151.8 149.7 149.3 147.8 143.6 124.6 115. 0 114.7 114.6 114.3 102. 0 (CHI), 100. 1 94.7 82.2 7 1. 1 7 1. 0 (CHI), 5 6.4 (CH3), 5 5.7 (CH3) 21.9 (CH3) 21.8 (CH3).MNS (70eV)m/z 3 70 (MI 3 55 M' CH3 328 (18, M CH(CH 3 3 27 M' CH2CHCH 3 2 86 (100, M+ 2 x CH2CHCH3) 271 (42, M -2 x CH2CHCH3 CH3). Anal. Calcd for C 22

H

26 0 5 C, 71.33; H, 7.07. Found: C, 70.91; H, 7.24.

J-(a-Iodoacetoxy)-2-[(4-isopropoxy-3methoxyphenyl)ethnyl]S5isopropoxy.4-methoybenzene: DCC (1.60 g, 7.75 mmol) was added to a solution of 2-iodoacetic acid (1.44 g, 7.74 mmol), 2- 4 -sopropoxy-3-methoxy-phenyl)ethyny1]-5-isopropoxy-4-metioxypheno (2.60 g, 7.03 mmol) and DMAP (43 mg, 0.35 mmol) in CH2C12 (30 mL) and the solution thereby obtained was stirred at 18*C for 3 h. The resulting suspension was filtered (CH2CI2 rinse) and the filtrate concentrated under reduced pressure. The residue was suspended in ether (30 mL) at 0 0 C with rapid stiffing then filtered [rinsing with Et2O (20 mL) pre-cooled to 0 0 C] giving the title compound (3 .67 g, 97%) as a cream solid mp 127-8*C. IR (KBr disc, cm-i) 2969, 2930, 2833, 1755, 1611, 1575, 1516, 1467, 1416, 1402, 1385, 1365, 1320, 1252, 1236, 1212, 1135, 1108. 1

H

NMR (300 MHz, CDCl 3 57.08 (dd, J= 1.8,8.4 Hz, 1H), 7.04 J= 2.1 Hz, 1H), 7.01 IR), 6.82 J= 8.4 Hz, 1H), 6.65 1H), 4.53 (septet, J= 6.0 Hz, 2Hz), 3.95 2H), 3.85 6HI), 1.3 8 J= 6.0 Hz, 6H), 1.37 J= 6.0 Hz, 6H). 13 C NMR APT (75.5 MIHz, CDCI 3 5 167.3 150.0 148.4 148.3 148.2 145.1 125.2 (CII), 115.6 115.3 (CII), 115.2 (CHI), 115.0 (CHI), 108.8 (CHI), 93.6 82.7 72.0 (CII), 71.6 (CHI), 56.5 (CH3) 56.3 (CH3), 22.3 (CH3), 22.0 (CH3), -6.2 (CH2). MIS (70 eV) m/z 538 (80, 496 (10, MI CH2CHCH 3 370 CH2ICO), 328 286 (100); fIRMS calcd for C24H 2 70 6 1 538.0852. Found 538.0854.

.WO 98/50365 PCT/AU98/00312 36 Lamellarin K triisopropyl ether: 3, 4 -Dihydro-6,7-dimethoxy-5-isopropoxyisoquinoline (1.20 g, 4.81 mmol) was added to a solution of 1-(a-iodoacetoxy)-2-[(4-isopropoxy-3-methoxyphenyl)ethynyl]-5-isopropoxy-4-methoxybenzene (2.30 g mg, 4.27 mmol) in dry 1,2dichloroethane (40 mL) and the solution stirred at 18"C for 8 h. After this time diisopropylethylamine (750 pL, 4.30 mmol) was added and the reaction mixture heated at 83 "C for 32 h. The reaction mixture was cooled, evaporated on to silica gel (6 g) and the residue subjected to flash chromatography on silica gel (sequential elution with 2:1:0, 2:3:1 hexane/CH2Cl2/ether) concentration of the appropriate fractions (Rf 0.4 5:5:2 hexane/CH2Cl2/ether) giving the title compound (2.28 g, 81 as a white solid mp 244-5"C.

IR(KBr disc, cm 1 2974, 2935, 2832, 1702, 1620, 1539, 1506, 1476, 1465, 1444, 1420, 1259, 1237, 1203, 1175, 1110, 1040. 1 H NMR (300 MHz, CDCI 3 6 7.10 2H), 7.05 1H), 6.92 1H), 6.64 1H), 6.60 1H), 4.74 (br t, J= 6.6 Hz, 2H), 4.56 3H), 3.83 6H), 3.42 3H), 3.34 3H), 3.15 (br t, J= 6.6 Hz, 2H), 1.41 J= 6.0 Hz, 6H), 1.39 J= 6.0 Hz, 6H), 1.31 J= 6.0 Hz, 6H). 13 C NMR APT (75.5 MHz, CDCl 3 5 155.6 151.7 151.2 148.5 147.0 146.9 146.5 145.9 142.5 135.5 128.5 128.1 123.3 123.0 121.1 116.8 115.5 114.5 113.8 110.3 104.9 104.8 104.7 103.4 75.7 71.7 71.4 (CH), 60.6 (CH3), 56.1 (CH3), 55.4 (CH3), 55.1 (CH3), 42.3 (CH2), 22.7 (CH3), 21.8 (CH3), 21.7 (CH3). MS (70 eV) m/z 657 (100, 615 (44, M* CH2CHCH3), 572 M* 2 x CH2CHCH3); HRMS calcd for C 38

H

43 N0 9 657.2938. Found 657.2938.

Example 3.

Lamellarin K (Compound 21) Lamellarin K: Aluminium chloride (1.33 g, 9.94 mmol) was added to a solution of lamellarin K triisopropylether (1.80 g, 2.76 mmol) in dry CH2C12 (20 mL) and the WO 98/50365 PCT/AU98/00312 -37reaction allowed to stir for 4 h. After this time the reaction mixture was treated with NH4Cl (a saturated solution in H20, 20 mL). The two phases were transfered to a separatory funnel, diluted with ethyl acetate (50 mL) and washed with H20 (40 mL). The aqueous layer was extracted with ethyl acetate (2 x 40 mL). The combined organic phases were dried (MgS04) and concentrated on to silica gel (8 The residue was subjected to flash chromatography on silica gel (sequential elution with 20:1, 10:1, 5:1 CH2Cl2/methanol) the relevant fractions (Rf0.6 10:1 CH2Cl2/methanol) were concentrated giving lamellarin K (1.4 mg, 95 as white solid, mp 230-2*C. IR (KBr disc, cm-1) 3472, 2940, 2839, 1709, 1600, 1549, 1511, 1458, 1428, 1407, 1265, 1207, 1142, 1122, 1031. 1

H

NMR (300MHz, CDCl 3 8 7.13 J= 8.1 Hz, 1H), 7.07 (dd, J= 1.5, 8.1 Hz, 1H), 6.97 1H), 6.96 J= 1.5 Hz, 1H), 6.59 1H), 6.38 1H), 5.95 1H), 5.75 1H), 5.71 1H), 4.90 1H), 4.64 1H), 3.89 3H), 3.87 3H), 3.49 3H), 3.36 3H), 3.12 2H). 1 HNMR (300 MHz, d6DMSO) 6 9.29 (br s, 2H), 7.03 (br s, 1H), 6.99 (dd, J= 1.5, 8.1 Hz, 1H), 6.82 (d,J 8.1 Hz, 1H), 6.74 1H), 6.54 1H), 6.34 1H), 4.54 2H), 3.75 3H), 3.55 3H), 3.35 3H), 3.27 3H), 3.01 2H). 13 C NMR+ APT (75.5 MHz, d6DMSO) 5 154.6 151.0 148.7 147.5 147.0 146.7 145.9 144.6 136.6 135.5 127.8 125.8 123.6 122.7 116.5 115.7 114.9 114.5 112.8 108.9 105.2 103.8 101.1 (CH) 60.5 (CH3), 56.2 (CH3), 55.2 (CH3), 54.8 (CH3), 41.9 (CH2), 21.5 (CH2). MS (70 eV) m/z 531 (100, M 516 (17, M CH3), 265.5 M 2 HRMS calcd for C 29

H

25 N0 9 531.1539. Found 531.1524.

Example 4.

Lamellarin T diisopropyl ether (Compound 37) P, P-Dibromo-3-isopropoxy-4-methoxystyrene: Carbon tetrabromide (51.3 g, 154.7 mmol) was added portion-wise to a magnetically stirred mixture of zinc dust (10.1 g, 154.5 .WO 98/50365 PCT/AU98/00312 -38mmol) and PPh3 (40.5 g, 154.4 mmol) in CH2C12 (350 mL) maintained at 0"C on an icesalt bath. The resulting suspension was allowed to warm to room temperature and then stirred for a further 22 h. After this time the reaction mixture was re-cooled to 0*C and isovanillin isopropyl ether (15.0 g, 77.3 mmol) was added dropwise over 2 min. The resulting mixture was allowed to stir at room temperature for 1 h then diluted with hexane (200 mL) and filtered through a No. 3 porosity sintered-glass funnel. The filtrate was concentrated on to silica gel (400-200 mesh, 30 g) and the resulting solid added to the top of a flash chromatography column (20 cm long x 10 cm wide) which was subjected to gradient elution with 2:1, 1:1 then 1:2 hexane/CH2Cl2 Concentration of the appropriate fractions (Rf 0.5 in 1:1 hexane/CH2Cl2) afforded the title compound (27.0 g, 99%) as white crystalline masses, m.p. 67-69"C. vmax (KBr) 3009, 2975, 2929, 1597, 1572, 1508, 1463, 1441, 1429,1373, 1301, 1276, 1260, 1233, 1144, 1110 and 999 cm-1. H n.m.r. 5 7.38 (s, 1H), 7.24 J 2.1 Hz, 1H), 7.08 (dd, J 8.4 and 2.1 Hz, 1H), 6.84 J 8.4 Hz, 1H), 4.53 (septet, J= 6.0 Hz, 1H), 3.86 3H), 1.38 J 6.0 Hz, 6H). 13 C n.m.r. 8 150.5 146.7 136.4 127.7 122.1 115.3 111.3 87.0 71.4 55.8 (CH3), 22.0 (2 X CH3). Mass spectrum m/z 352 (20) 350 (40) 348 (22) (M 310 (50) 308 (100) 306 (50) C3H6) 295 293 (61) 291 (33) C3H6 CH3 Anal. Calcd for C 12

H

14 Br 2

O

2 C, 41.2; H, 4.0; Br, 45.7. Found: C, 41.1; H, 3.7: Br, 45.3%.

2 3 -Isopropoxy- 4 -methoxy-phenyl)ethynyl]-5-isopropoxy-4-methoxybenzaldehyde: n- Butyllithium (45.8 ml of a 2.5 M solution in hexane, 114.5 mmol) was added dropwise over 0.2 h to a magnetically stirred solution of 3P,-dibromo-3-'isopropoxy-4methoxystyrene (20.0 g, 57.2 mmol) in THF (250 mL) maintained at -78 0 C on a dry-ice acetone bath. The slightly tan coloured solution thereby obtained was stirred at -78"C for h then the reaction vessel was removed from the cooling bath. After 0.1 h anhydrous ZnCl2 (7.79 g, 57.2 mmol dried at 120*C and 0.01 mm Hg for 20 h) was added to the reaction mixture which slowly became colourless as it was warmed to room temperature over 1 h. Aldehyde (17.8 g, 55.6 mmol) then PdCI2(PPh3)2 (195 mg, 0.128 mmol) were added and the reaction mixture stirred at 18 C for 6 h. After this time the reaction mixture WO 98/50365 PCT/AU98/00312 -39was diluted with ethyl acetate (600 mL) and washed with water (2 x 500 mL) then dried (MgSO4), filtered and concentrated under reduced pressure. The solid thereby obtained was suspended in ether/hexane (100 ml of a 4:1 v/v mixture) and the resulting suspension was subjected to vacuum filtration and the solid thus retained was washed with ether/hexane (2 x 50 ml of a 4:1 v/v mixture) to afford the title compound (16.5 g, 78%) as white crystalline masses, mp 110-113 OC. Subjection of the combined filtrates to flash chomatography (silica, 1:1, 1:2 hexane/CH2Cl2, CH2C12 and then CH2C12 gradient elution). Concentration of the appropriate fractions (Rf 0.4 in CH2C12) then afforded additional quantities of product (3.64 g, 14 mp 110-113 OC; vmax (KBr) 2978, 2933, 2837, 2203, 1685, 1589, 1515, 1508, 1448, 1397, 1387, 1376, 1358, 1322, 1276, 1240, 1216, 1156, 1132 and 1091 cm- 1 1 Hn.m.r. 6 10.49 1H), 7.41 1H), 7.21 (dd, J= 8.4 and 2.4 Hz, 1H), 7.06 J= 2.4 Hz, 1H), 7.03 1H), 6.84 J= 8.4 Hz, 1H), 4.70 (septet, J= 6.0 Hz, 1H), 4.57 (septet, J= 6.0 Hz, 1H), 3.97 3H), 3.88 3H), 1.40 (2 x d, J= 6.0 Hz, 2 x 6H); 13 C n.m.r. 6 190.5 154.5 151.2 147.8 146.9 129.7 125.2 121.5 118.2 114.4 111.7 110.7 (CH), 95.0 83.4 71.4 71.1 56.1 (CH3), 55.8 (CH3), 21.9 (CH3), 21.8 (CH3); Mass spectrum m/z 382 (3 298 [100, (M 2 x H2CCHCH3) 283 [46, (M 2 x H2CCHCH3, HRMS, Calcd for C 23

H

26 0 5 382.1780. Found 382.1781.

2 3 -Isopropoxy-4-methoxy-phenyl)ethynyl]5-isopropoxy-4-methoxyphenol: m- Chloroperoxybenzoic acid [19.4 g, ALDRICH, 50% (remainder 3-chlorobenzoic acid and water), ca. 56 mmol] was added in portions over 0.25 h to a magnetically stirred mixture of 2 3 -isopropoxy-4-methoxy-phenyl)ethynyl]-5-isopropoxy-4-methoxybenzaldehyde (18.0 g, 47.1 mmol) and KHCO3 (14.1 g, 141.0 mmol) in CH2C12 (220 ml) maintained at 0"C (ice-bath). The resulting slurry was stirred for a further 1 h between 0"C and 18"C then filtered through Celite® and the solids thus retained rinsed with CH2C12 (1 x 200 mL). The combined filtrates were concentrated under reduced pressure and the residue treated with NH3 (1 x 300 mL of a saturated methanolic solution). After 1 h at 18 C the reaction mixture was concentrated under reduced pressure and the residue redissolved in CH2C12 (200 ml) then concentrated on to silica gel (20 g, 400-200 mesh). The resulting WO 98/50365 PCT/AU98/00312 powder was loaded on top of a column of silica (40-10 mesh TLC grade, 10 cm wide x cm long) and eluted with 2:1, 1:1 and 1:2 hexane/CH2Cl2 then neat CH2C12 (250 mL of each). The appropriate fractions (Rf 0.3 in 1:2 hexane/CH2Cl2) were concentrated under reduced pressure to give the title compound (16.2 g, 93%) as white crystalline masses, m.p. 139-140"C. vmax (KBr) 3431, 2983, 2934, 1513, 1461, 1332, 1267, 1241, 1210, 1136, 1110, 1024 and 991 cm- 1 1 H n.m.r. 6 7.12 (dd, J 8.1 and 2.1 Hz, 1H), 7.10 (d, J 2.1 Hz, 1H), 6.88 1H), 6.84 J 8.1 Hz, 1H), 6.56 1H), 5.61 1H), 4.55 (septet, J 6.0 Hz, 2 x 1H), 3.88 3H), 3.82 3H), 1.38 (2 x d, J 6.0 Hz, 2 x 6H).

13C n.m.r. 5 151.8 151.1 149.6 147.0 143.9 125.1 118.4 114.8 114.2 111.7 101.9 100.0 95.2 81.8 71.6 71.2 56.7 (CH 3 56.0 (CH3), 22.1 (CH3), 21.9 (CH3). Mass spectrum m/z 370 (68, 328 [24, (M H2CCHCH 3 286 [70, (M 2 x H2CCHCH 3 271 [100, 2 xH2CCHCH 3 Anal. Calcd for C 22

H

26 0 5 C, 71.3, H, 7.1. Found: C, 71.5, H, 7.1.

1-(oc-lodoacetoxy)- 2 3 -isopropoxy-4-methoxy-phenyl)ethynyl]-5-isopropoxy-4methoxybenzene: DCC (6.43 g, 31.1 mmol) was added to a solution of 2-iodoacetic acid (5.80 g, 31.2 mmol), 2-[(3-isopropoxy-4-methoxy-phenyl)ethynyl]-5-isopropoxy-4methoxyphenol (11.0 g, 29.7 mmol) and DMAP (36 mg, 0.30 mmol) in CH2C12 (200 mL) and the solution thereby obtained was stirred at 18 C for 3 h. The resulting suspension was filtered (CH2C1 2 100 mL rinse) and the filtrate concentrated under reduced pressure. The residue thus obtained was suspended in ether (150 mL) at 0C with rapid stirring for 2 h then filtered [rinsing with Et20 (70 mL) pre-cooled to 0 C] giving the product (15.0 g, 94 as a cream solid mp 136-7"C. IR (KBr disc, cm-1) 3021, 2978, 2930, 1771, 1607, 1512, 1460, 1444, 1420, 1330, 1241, 1211, 1133, 1110, 1096. 1 H NMR (300 MHz, CDC1 3 7.12 (dd, J= 1.8, 8.4 Hz, 1H), 7.06 J= 2.1 Hz, 1H), 7.02 1H), 6.82 J= 8.4 Hz, 1H), 6.66 1H), 4.55 (septet, J= 6.0 Hz, 2Hz), 3.96 2H), 3.87 6H), 1.40 J= Hz, 6H), 1.38 J= 6.0 Hz, 6H). 13 C NMR APT (75.5 MHz, CDC13) 6 166.9 150.8 147.9 147.8 146.7 144.6 125.1 118.3 115.0 114.7 111.5 108.3 93.2 82.3 71.5 71.3 56.1 WO 98/50365 PCT/AU98/0031 2 -41- (CH3), 55.7 (CH3), 22.0 21.8 (CH3), -6.6 (CH 2 MIS (70 eV) m/z 53 8 (100, 496 (12, M+ CH2CHCH3), 370 (26, M+ CH2ICO), 328 286 (8 HRMS calcd for C 24

H

27 0 6 1 538.0852. Found 538.0843.

Lamellarin Tdiisopropylether: 3 4 -Dihydro-5,6,7-trimethoxyisoquinoline (986 mg, 4.46 mmol) was added to a solution of l-(ct-iodoacetoxy)-2-[(3-isopropoxy-4-methoxyphenyl)ethynyl]-5-isopropoxy-4-methoxybenzene (2.0 g, 3.71 mmol) in dry 1,2dichloroethane (50.0 mL) and the solution stirred at 18*C for 15 h. After this time diisopropylethylamine (677 gL, 3.90 mmol) was added and the reaction mixture heated at 83 *C for 24 h. The reaction mixture was cooled, evaporated on to silica gel (5 g) and the residue subjected to flash chromatography on silica gel (elution with 9:1, 6:1 CH2CI2/ether) concentration of the appropriate fractions (Rf 0.5 7:1 CH2CI2/ether) giving the title compound(l.67 g, 71 as a white solid, xnp 192-5*C. JR (KBr disc, cm-1) 2975, 2935, 2834, 1700, 1620, 1540, 1506, 1476, 1455, 1442, 1416, 1259, 1239, 1208, 1175, 1137, 1116, 1084, 1037, 1013. IH NMvR (300 Mffz, CDC1 3 8 7.07 (br s, 2H), 7.03 III), 6.89 I 6.63 I 6.5 6 I1H), 4.74 (br t, J 6.6 Hz, 2H), 4.5 2 (in, 2H), 3.92 (s, 3H), 3.88 311), 3.85 311), 3.42 3H), 3.3 3 3H), 3.12 (br t, J 6.6 Hz, 2H), 1.3 6 J =6.0 Hz, 6H), 1.32 J 6.0 Hz, 6H4). 1 3 C NMvR APT (75.5 M&z, CDCl 3 8 155.6 151.8 150.6 150.0 148.0 147.0 146.5 145.9 142.1 135.3 128.1 127.8 123.6 123.0 120.0 117.7 (CH), 115.5 113.8 112.6 110.3 105.1 104.8 103.3 (CHI), 71.3- 71.2 (CHI), 61.0 (CH3), 60.5 (CH3), 56.3 (CH3), 55.4 (CH3), 55.1 (CH3), 42.1 (CH2), 22. 0 (CH3), 21.9 (CH2), 21.8 (CH3). MS (70 eV) m/z 629 (100, M, 587 (63, M+ CH2CHCH 3 HRMS calcd for C 36 H4 36 N0 9 629.2625. Found 629.2642.

Example Lamellarin T (Compound 26) Lamellarin T: Aluminium chloride (480 mg, 3.60 nunol) was added to a solution of lamellarin T diisopropylether (945 mng, 1.50 inmol) in dry CH2C1 2 (10.0 mL) and the WO 98/50365 PCT/AU98/00312 -42reaction allowed to stir for 0.5 h. After this time the reaction mixture was treated with NH4CI (a saturated solution in H20, 5 mL). The two phases were transfered to a separatory funnel, diluted with H20 (100 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic phases were dried (MgSO4) and concentrated on to silica gel (4 The residue was subjected to flash chromatography on silica gel (sequential elution with 99:1, 20:1 CH2Cl2/methanol) the relevant fractions (Rf 0.2 20:1 CH2Cl2/methanol) were concentrated giving lamellarin T (736.8 mg, 90 as white solid, mp 283-4"C. IR (KBr disc, cm- 1 3492, 3303, 2998, 2982, 2835, 1677, 1624, 1582, 1551, 1507, 1476, 1462, 1450, 1413, 1273, 1249, 1207, 1160, 1122, 1041, 1023.

1H NMR (300 MHz, d6DMSO) 6 9.65 1H), 9.29 1H), 7.13 J 8.7 Hz, 1H), 6.90 2H), 6.80 1H), 6.65 1H), 6.60 1H), 4.65 1H), 4.56 1H), 3.83 3H), 3.79 3H), 3.75 3H), 3.38 3H), 3.30 3H), 3.06 (br t, J= 7.0 Hz, 2H). 13 C NMR+ APT (75.5 MHz, d6DMSO) 8 154.3 151.3 150.2 147.7 147.5 146.8 145.6 144.4 141.8 134.5 127.3 127.1 122.4 121.5 120.0 117.7 115.2 113.3 112.8 108.5 105.1 104.9 103.6 60.8 (CH3), 60.5 (CH3), 56.0 (CH3), 55.0 (CH3), 54.7 (CH3), 41.6 (CH2), 21.4 (CH2): MS (70 eV) m/z 545 (100, M 530 (31, M't CH3) 272.5 (27, M 2 HRMS calcd for C 30

H

2 1 NO9 545.1686.

Found 545.1690.

Example 6.

Lamellarin T diacetate (Compound 39) Lamellarin Tdiacetate: Lamellarin T (48 mg, 0.088 mmol) was dissolved in a solution of acetic anhydride (1.0 mL) and pyridine (1.0 mL) containing DMAP (several crystals) and the solution stirred at 18"C for 26 h. The solution was then diluted with ethyl acetate (15 mL) and washed with NaHCO3 (a saturated solution in H20, 20 mL) and citric acid (10% in H20, 20 mL) dried over (MgSO4) and concentrated under reduced pressure. The residue was subjected to flash chromatograophy on silica gel (sequential elution with 2:1, 1:1 hexane/ethyl acetate) the relevant fractions (RJ 0.42 1:1 WO 98/50365 PTA9/01 PCT/AU98/00312 -43hexane/ethyl acetate) were concentrated giving the title compound (47 mg, 83 as a white solid mp 251-2*C. 1k (KBr disc, cm-i) 2937, 2840, 1769, 1718, 1603, 1545, 1507, 1475, 1456, 1414, 1295, 1266, 1201, 1141, 1117, 1036. 1 H NMvR (300 Iv&~, CDC13) 6 7.3 1 (dd, J 8.4 Hz, 1H), 7.22 J 1 Hz, I1H), 7.14 J 8.7 Hz, 111), 7.07 111), 4.90 (in, III), 4.60 (in, 1H), 3.90 3H), 3.89 3H), 3.86 3H), 3.45 314), 3.39 311), 3.13 (in, 2H), 2.30 6H4). 13 C NMIR+ APT (75.5 M&, CDCI3) 5 168.7 168.4 155.0 151.9 147.5 144.8 142.2 140.6 13 8.7 13 5.6 129.5 127.6 127.2 125.4 122.6 119.8 116.1 114.8 114.4 112.9 (CHI), 111.7 105.4 (CH), 105.0 (CMI, 60.9 (CH3), 60.8 (CH3), 56.2 (CH3), 55.5 (CH3), 55.2 (CH3), 42.1 (CH2), 21.7 (CH2), 20.4 (CH3). MS (70 eV) m/z 629 (53, MI 587 (100, M CH2CO); HRMS calcd for C 34

H

31 N0 11 629.1897. Found 629.1907.

Example 7.

10-Deoxylamellarin K diisopropylether (Compound 34) JO-Deoxylamellarin K disopropylether: 3 ,4-Dihydro-6,7-dimethoxyisoquinoline (170 mg, 0.82 inmol) was added to a solution of 1-(a-iodoacetoxy)-2-[(4-isopropoxy-3inethoxy-phenyl)ethynyl]-5-isopropoxy-4-inethoxybenzene (400 mg, 0.74 mmol) in dry 1,2-dichioroethane (4.0 mL) and the solution stirred at 18TC for 5 h. After this time diisopropylethylamine (13 6 pLL, 0.78 iniol) was added and the reaction mixture heated at 83 C for 28 h. The reaction mixture was cooled, evaporated on to silica gel (3 g) and the residue subjected to flash chromatography on silica gel (sequential elution with 1:2:0, 3:6:1, 0:5:1 hexane/CH2Cl2/ether) concentration of the appropriate fractions (Rj 0.7 9:1 CH2C12/ether) giving the title compound (352 ing, 79 as a white solid, mp 222-3 IR(KBr disc, cm- 1 2975, 2933, 2831, 1709, 1611, 1578, 1543, 1514, 1485, 1464,1438,1415,1271,1240,1212, 1165, 1042. 1 HNMR (300v11{z, CDC1 3 5 7.11 (in, 2H), 7.05 1H1), 6.92 1H), 6.77 1H), 6.75 114), 6.68 11H), 4.80 (mn, 2H), 4.57 (mn, 214), 3.90 31H), 3.83 311), 3.43 3H), 3.37 314), 3.12 (br t, J 7.0 Hz, 2H), 1.41 J =6.0 Hz, 614), 1.39 J 6.0 Hz, 6H1). 13 C NMR APT WO 98/50365 PCT/AU98/00312 -44- (75.5 MHz, CDC1 3 5 155.5 151.2 148.9 147.4 147.0 146.9 146.5 146.0 135.9 128.5 128.3 126.6 123.4 120.1 116.8 114.9(C), 114.5 113.7(C), 110.9 110.3 108.6 (CH), 104.8 103.4 71.7 71.4 56.2 (CH3), 55.9 (CH3), 55.5 (CH3), 55.1 (CH3), 42.4 (CH2), 28.7 (CH2), 21.9 (CH3), 21.8 (CH3). MS (70 eV) m/z 599 (100, M 557 (61, M CH2CHCH3), 515 (49, h 2 x CH2CHCH3); HRMS calcd for C 35

H

37 NOg 599.2519. Found 599.2519.

Example 8.

10-Deoxylamellarin K (Compound K: Aluminium chloride (80.3 mg, 0.60 mmol) was added to a solution of 10-deoxylamellarin K diisopropylether (120 mg, 0.20 mmol) in dry CH2C12 (10.0 mL) and the reaction allowed to stir for 1 h. After this time the reaction mixture was treated with NH4Cl (a saturated solution in H20, 10 mL). The two phases were transfered to a separatory funnel, diluted with ethyl acetate (40 mL) and washed with (40 mL). The aqueous layer was extracted with ethyl acetate (2 x 20 mL). The combined organic phases were dried (MgSO4) and concentrated on to silica gel (2 g).

The residue was subjected to flash chromatography on silica gel (sequential elution with 20:1, 10:1 CH2Cl2/methanol) the relevant fractions (Rf 0.7 10:1 CH2Cl2/methanol) were concentrated giving 10-deoxylamellarin K (91.7 mg, 89 as white solid mp, 292-4°C. IR (KBr disc, cm- 1 3529, 3105, 3003, 2937, 2833, 1667, 1609, 1546, 1520, 1486, 1464, 1439, 1416,1274,1217, 1163, 1047. 1HNMR (300 MHz, CDC1 3 6 7.13 J= 8.1 Hz, 1H), 7.08 (dd, J= 1.5, 8.1 Hz, 1H), 6.98 J= 1.5 Hz, 1H), 6.96 (s, 1H), 6.76 1H), 6.71 1H), 6.64 1H), 5.76 1H), 5.74 1H), 4.96 1H), 4.64 1H), 3.90 3H), 3.87 3H), 3.51 3H), 3.38 3H), 3.11 2H). 1H NMR (300 MHz, d6DMSO) 5 9.40 1H), 9.09 1H), 6.99 J 8.1 Hz, 1H), 6.96 1.5 Hz, 1H), 6.85 (dd, J= 1.5, 8.1 Hz, 1H), 6.82 1H), 6.74 1H), 6.67 (s, 1H), 6.59 1H), 4.62 2H), 3.78 6H), 3.39 3H), 3.27 3H), 3.05 (br t, J =7.0 Hz, 2H). 13 CNMR+APT (75.5 MHz, d6DMSO) 6 152.8 147.0 146.7 .WO 98/50365 PCT/AU98/00312 45 145.3 145.0 144.9 144.1 142.7 133.8 126.2 124.8 123.9 121.8 117.9 114.5 113. 0 112.8 1 11. 0 109.7 107.3 107.0 103.4 (CHi), 101.9 54.3 (CH3), 53.8 (CH3), 53.4 (CH3), 52.9 (CH3), 40.3 (CH2), 26.3 (CH2). MS (70 eV) m/z 515 (100, M-' 257.5 (19, M 2 HRMS calcd for C 29

H

25 N0 8 515.1580. Found 515.1576.

Example 9.

Lamellarin U diisopropylether (Compound 38) Lamellarin U disopropylether:- 3,4-Dihydro-6,7-dirnethoxyisoquinoline (425 mg, 2.23 mmol) was added to a solution of 1-(a-iodoacetoxy)-2-[(3-isopropoxy-4-methoxyphenyl)ethynyl]-5-isopropoxy-4-methoxybenzene (1.00 g, 1.86 mmol) in dry 1,2dichioroethane (30.0 mL) and the solution stirred at I8*C for 17 h. After this time diisopropylethylamine (340 pL, 1.95 mmol) was added and the reaction mixture heated at 83 *C for 20 h. The reaction mixture was cooled, evaporated on to silica gel (5 g) and the residue subjected to flash chromatography on silica gel (elution with 9:1 CH2Cl2/ether) concentration of the appropriate fractions (Rf 0.5 7:1 CH2Cl2/ether) gave the title compound (780 mg, 70 as a white solid, mp 213 -4 IR (KBr disc, cm-i) 2975, 2936, 2836, 1694, 1620, 1608, 1580, 1542, 1511, 1485, 1463, 1440, 1415, 1271, 1259, 1241, 1213, 1167, 1043. 1 HNMiR(3 00 CDCI 3 8 7. 01 (br s, 2H), 7.03 1H), 6.87 1H1), 6.74 1H1), 6.69 1H), 6.65 1H), 4.75 (in, 2H1), 4.50 (in, 2H), 3.90 3H), 3.86 311), 3.41 3H), 3.3 5 3H), 3. 10 (br t, J =6.9 HIz, 211), 1.34 J= 6.0 Hz, 6H), 1.32 J= 6.0 Hz, 611). 13 C INMR APT (75.5 CDCl 3 5 155.6 149.9 148.8 147.9 147.4 146.9 146.4 145.9 135,8 128.2 127.8 126.5 123.6 120.0 117.7 114.8 113.6 112.5 (CHi), 110.9 110.3 108.5 104.8 103.3 71.3 71.2 56.3 (CH3), 55.9 (CH3), 55.5 (CH3) 55.1 (CH3), 42.3 (CH2), 28.6 (CH2), 21.9 (CH3), 21.8 (CH3). MS (70 eV) m/z 599 (100, M+ 557 (8 1, M+ CH2CHCH3), 515 (41, M* 2 x CH2CHCH3); HRMS calcd for CH 35

H

37 N0 8 599.2519. Found 599.2526.

WO 98/50365 PCT/AU98/00312 -46- Example Lamellarin U (Compound 29) Lamellarin U: Aluminium chloride (315.8 mg, 2.36 mmol) was added to a solution of lamellarin U diisopropylether (430.0 mg, 0.717 mmol) in dry CH2C12 (20.0 mL) and the reaction allowed to stir for 14 h. After this time the reaction mixture was treated with NH4Cl (a saturated solution in H20, 10 mL). The two phases were transfered to a separatory funnel, diluted with ethyl acetate (40 mL) and washed with H20 (20 mL).

The aqueous layer was extracted with ethyl acetate (2 x 20 mL). The combined organic phases were dried (MgSO4) and concentrated on to silica gel (2 The residue was subjected to flash chromatography on silica gel (sequential elution with 20:1, 10:1 CH2Cl2/methanol) the relevant fractions (Rf 0.7 10:1 CH2Cl2/methanol) were concentrated giving lamellarin U (347.1 mg, 94 as white solid, mp 242-3 OC. IR (KBr disc, cm 1 3526, 3449, 3296, 3001, 2954, 2838, 1683, 1674, 1609, 1584, 1485, 1464, 1440, 1413, 1273, 1247, 1215, 1171, 1048. 1 H NMR (300 MHz, d6DMSO) 6 9.67 1H), 9.30 1H), 7.15 J 8.1 Hz, 1H), 6.98 1H), 6.90 2H), 6.80 1H), 6.69 2H), 4.62 2H), 3.84 3H), 3.78 3H), 3.39 3H), 3.26 (s, 3H), 3.11 (br t, J= 7.0 Hz, 2H). 13 C NMR+ APT (75.5 MHz, d6DMSO) 6 154.6 149.2 148.0 147.9 147.3 146.0 144.8 135.7 127.7 127.6 127.3 122.0 119.6 118.2 114.7 113.8 112.9 112.1 ,109.0 108.9 105.4 103.4 56.4 (CH3), 55.9 (CH3), 55.4 (CH3), 54.8 (CH3), 42.9 (CH2), 28.0 (CH2). MS (70 eV) m/z 515 (100, 257.5 (15, M 2 HRMS calcd for C 28

H

25

NO

8 515.1580. Found 515.1586.

Example 11.

Lamellarin W diisopropylether (Compound 11) Lamellarin W diisopropylether: DDQ (219 mg 0.963 mmol) was added to a solution of lamellarin T diisopropyl ether (485 mg, 0.77 mmol) in dry chloroform (10 mL) and the reaction stirred at 61 C for 2 h. The reaction mixture was cooled, evaporated on to WO 98/50365 PCT/AU98/00312 -47silica gel (3 g) and the residue subjected to flash chromatography on silica gel (sequential elution with(9:l, 4:1 CH2Cl2/ether) concentration of the appropriate fractions (Rf0.6 6:1 CH2Cl2/ether) gave the title compound (479 mg, 99 as a white solid, mp 200-1"C. IR (KBr disc, cm-1) 2975, 2935, 2834, 1698, 1621, 1606, 1536, 1498, 1480, 1453, 1429, 1418, 1395, 1260, 1233, 1177, 1137, 1117, 1073, 1045, 975.

1 H NMR (300 MHz, CDCl 3 6 9.21 J= 7.8 Hz, 1H), 7.38 J= 7.8 Hz, 1H), 7.17 (br s, 2H), 7.14 1H), 7.01 1H), 6.97 1H), 6.72 1H), 4.58 2H), 4.03 (s, 3H), 3.97 3H), 3.94 3H), 3.46 6H), 1.41 J= 6.3 Hz, 6H), 1.36 J= Hz, 6H). 1 3 C NMR APT (75.5 MHz, CDCI 3 6 155.5 153.2 150.2 148.4 1487.2 147.9 146.6 146.5 142.6 133.8 129.3 128.1 124.0 122.8 121.3 119.3 114.1 112.7 (CH), 111.9 109.8 108.1 106.8 105.4 103.3 101.5 71.3 71.2 61.6 (CH3), 61.1 (CH3), 56.4 (CH3), 55.4 (CH3), 55.1 (CH3), 21.9 (CH3), 21.8 (CH3). MS (70 eV) m/z 627 (100, M 585 (85, l CH2CHCH3); HRMS calcd for CH 36

H

37 N0 9 627.2468. Found 627.2475.

Example 12.

Lamellarin W (Compound 9) Lamellarin W: Aluminium chloride (112 mg, 0.836 mmol) was added to a solution of lamellarin W diisopropylether (175 mg, 0.279 mmol) in dry CH2C12 (10.0 mL) and the reaction allowed to stir for 14 h. After this time the reaction mixture was treated with NH4CI (a saturated solution in H20, 5 mL). The two phases were transfered to a separatory funnel, diluted with H20 (40 mL) and extracted with ethyl acetate (3 x 40 mL) and washed.

The combined organic phases were dried (MgSO4) and concentrated on to silica gel (2 g).

The residue was subjected to flash chromatography on silica gel (sequential elution with 99:1, 20:1 CH2Cl2/methanol) the relevant fractions (Rf0.2 20:1 CH2Cl2/methanol) were concentrated giving lamellarin W (143 mg, 94 as white solid, mp 284-6*C. IR (KBr disc, cm-1) 3413, 3135, 2937, 2841, 1667, 1606, 1481, 1424, 1275, 1240, 1207, 1156, 1075, 1045. 1 H NMR (300 MHz, d6DMSO) 6 9.50 (br s, 2H), 8.89 J= 7.8 Hz, 1H), NWO 98/50365 PTA9/01 PCT/AU98/00312 -48 7.28 J 7.8 Hz, 1H1), 7.09 J 8.1 Hz, 111), 6.96 2H), 6.87 J 8.1 Hz, 1H'),6.82 1H), 6.67 lH), 3.92 311), 3.84 311), 3.81 3H), 3.37 6H). 1 3

C

NMvIR APT (75.5 MfHz, d6DMSO) 5 154.8 152.4 147.5 147.1 146.9 146.8 146.0 143.9 141.4 133.0 128.8(C), 127.4 121.9 121.8 120.6 118.4 117.7 (CHi), 111. 7 1 11. 3 10 8.3 107.7 106.0 104.8 103.3 (CHI), 101.2 61.0 (CH3), 60.3 (CH3), 55.6 (CH3), 54.7 (CH3), 54.4 (CH3). MS (70 eV) m/z 543 (100, M' 271.5 (11, M 2 HRMS calcd for 30

H

1 9 N0 9 543.1529. Found 543.1533.

WO 98/50365 WO 9850365PCT/AU98/00312 -49 Table 1 Compound R 1

W

2

R

3

R

4

R

5

R

6 1 H H H H H H 2(Lamellarin B) OMe OMe OMe H H OMe.

3(Lamellarin D) H OH OMe H H OMe 4(Lamellarin D- H OAc OMe H H OMe triacetate) M) OHf OMe OMe H H OMe 6(Lamellarin M- OAc OMe OMe H H OMe triacetate)I 7(Lamellarin N) H OH OMe H H OMe 8(Lamellarmn N- H GAc OMe H H OMe triacetate) 9(Lamellarin W) OMe OMe OMe IH H OMe 1O(Lamellarin X) OH OMe IOMe H H OMe I1I OMe IOMe OMe H H OMe' WO 98/50365 WO 9850365PCT/AU98/00312 Table 1. Continued Compound R 7

R

8

R

9 R1 0

R

1 R12 R1 3 1H H H H H H H 2(Lamellarin B) OHf H H OMe OH H H 3(Lamellarin D) OH H H OMe OH H H 4(Lamellarin D- OAc H H OMe OAc H H triacetate)

H

M) OHc H H OMe OHc H H 6(Lamellarin M- OHc H H OHe O~c H H triacetate)

H

7O(Lamellarin N) OH H H OH OMe H H 8(amllrr N-I~ H H O'Pc OMe H H WO 98/50365 WO 9850365PCT/AU98/00312 -51 Table 2.

Compound R1 R 2

R

3 R 4 R' R 6 13 (Lamellarin C) OMe OMe OMe H H OMe 14 (Lamellarin E) OH OMe OMe H H OMe F) OH OMe OMe H H OMe 16 (Lamellarin G) H OH OMe H H OH 17 (Lamellarin H) H OH OH H H OH 18 (Lamellarin 1) OMe OMe OMe H H OMe 19 (Lamellarin I- OMe OMe OMe H H OMe acetate) (Lamellarin J) H OH OMe H H OMe 21 21 (Lamellarm K) OHc OMe OMe H H OMe 22 (Lamellarin K- H~ OHe OMe H H OMe triacetate) 23 (Lamellarin L) H OH OMe H H OHe (Lamellarin S) H~ OHe OMe H H IOH4] 27 (Lamellarin OMe OMe OMe H H OMe

I-

.WO 98/50365 WO 9850365PCT/AU98/00312 -52- Table 2. Continued Compound R 7

R

8

R

9

R

10

R

1 R 12

R

13 R 1 4 12 (Lam 12 (Lamellarin A) OH H H OMe OH H H OH OH H H OMe OHe H H H 16 (Lam 14 (Lamellarin H) OH H H OH OHe H H H 18 (Lam 195(Lamellarin F) OHc H H OMe OMe H H H 16 (Lamellarin J) OHe H H OHe OMe H H H 217(LamellarinHK) OH H H OHe OH H H H 238(Lamellarin L) OH H H 0~ OMe H H H 19 (Lamellarin L- OAc H H O~e OMe H H H tracetate)I (Lamellarin T) OH H H OHe OMe H H H 22 (Lamellarin K2- OS03 H H OMe OHc H H H tsuaceate)

N

WO 98/50365 PCT/AU98/00312 -53- Table 2. Continued Compound R 7

R

8

R

9

R

10

R

11

R

12

R

13

R

14 28 H H H H H H H H 29 (Lamellarin U) OH H H OH OMe H H H (Lamellarin U20- OS03 H H OH OMe H H H sulfate) Na 31 (Lamellarin V) OH H H OH OMe H H OH 32 (Lamellarin V20- OSO3 H H OH OMe H H OH sulfate) Na 33 (Lamellarin Y20- OSO3 H H OH OMe H H H sulfate) Na 34 O'Pr H H OMe OiPr H H H OH H H OMe OH H H H 36 O'Pr H H OMe OiPr H H H 37 O'Pr H H OiPr OMe H H H 38 O'Pr H H O'Pr OMe H H H 39 (Lamellarin T OAc H H OAc OMe H H H diacetate) Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modification other than those specifically described. It is to be understood that the invention includes all such variations and modifications.

Claims (33)

1. A method for the preparation of a compound of general Formula A Zn (I) 0 W S comprising the step of cyclizing an azomethine ylide of general Foimula (II)! A Z. W o 20 wherein, heterocyclic group; or A is a cyclic group RARA 2 C-CRA3RA4 wherein RA2 and RA3, together with the carbon atoms to which they are attached form an optionally substituted saturated or unsaturated carbocyclic or heterocyclic group and RA' and RA 4 are as defined below or together form a bond; or A is a non-cyclic group RARAC-CRA3RA4 wherein RA RA 4 are as defined below and RI and RA 3 may optionally together form a bond; Z is a carbon or a heteroatom; P:\OPER\MJCM70170-98 .spl 299doc-26/I/Ol n is selected from 0, 1, 2 or 3; and RA'-A 4 and Y may be the same or different and each are selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally protected hydroxy, optionally substituted amino, optionally substituted alkoxy, optionally substituted alkenoxy, optionally substituted alkynoxy, optionally substituted aryl, optionally substituted heterocyclyl, carboxy, carboxy ester, carboxamido, acyl, acyloxy, mercapto, optionally substituted alkylthio, halogen, nitro, sulfate, phosphate and cyano, and W and X, together with the nitrogen and carbon atoms to which they are attached, form an isoquinolinyl or dihydroisoquinolinyl group which may be optionally S 10 substituted; or pharmaceutically acceptable derivatives and salts, racemates, isomers and/or tautomers thereof.

2. A method according to claim 1, wherein A is selected from benzene, naphthalene, pyridine, furan, pyrrole, thiophene, quinoline, indole, benzofuran and benzothiophene, each of which may be optionally substituted.

3. A method according to claim 2, wherein A is optionally substituted benzene. 20 4. A method according to claim 1, wherein A is a cyclic group RAlRA2C- SCRA3R A 4 wherein RA2-R A3 together with the carbon atoms to which they are attached form a cyclic group selected from cyclopentane, cyclopentene, cyclohexane, cyclohexene, cyclopentadiene, cyclohexadiene, tetrahydrofuran, dihydrofuran, pyrrolidine, pyrroline, pyran, dihydropyran, tetrahydropyran and piperidene, each of which may be optionally substituted. A method according to claim 1, wherein A is a non-cyclic group RARA2C- CRA3RA 4 wherein RA-R A4 are independently selected from hydrogen, PMOPERWMJI70170-98 rrpi 299.dc29/10/0I -56- optionally protected hydroxy, optionally substituted alkyl and optionally substituted alkoxy.

6. A method according to claim 5, wherein A is -H 2 C-CH 2

7. A method according to claim 1, wherein A is -HC=CH-.

8. A method according to claim 7, wherein W and X, together with the respective nitrogen and carbon atoms to which they are attached, form a group of Formula S a wherein ==is an optional double bond and R'-R 4 and R 14 are as defined for RAlA 4 in claim 1.

9. A method according to claim 8, wherein R'-R 4 and R 14 are selected from 20 hydrogen, hydroxy, optionally substituted alkyl, acyl and sulfate. A method according to claim 1, wherein n is 1.

11. A method according to claim 1, wherein Z is CH 2 nitrogen, sulfur or oxygen. 57

12. A method accordigi to claim wherein- Z is oxygen. 1-3. A method according to claim 1, wherein Y is an optionally substituted phienyl group. of Formula (ii) 00~0S 00 0 S *0 0 @00f 0 Q0~ a a 0< 0 0*5* @9 ~6 0 000050 0*0., S S 0 0**0 0 S OSSO a 000**a a S *000 5 a 00 00 a a S wherein are as defined for R'-Ri and R 14 in claim 8.

14. A method according to claim .13, wherein:,R? RP 3 are selected. from hydrogen, hydroxy, optionally substitutedl alkyl, optionally substituted alkoxy, and acyloxy.;.

15. A method according to claim 14, wh .erein 'the R 9 R 3are selected from hydrogen, hydroxy, methoxy, isopropoxy, methyl and acetoxy. 20 16. A method for the preparationdo acmpudoFrul(I) RS/\ Y R 8 II \(Ia) the step of cyclizing an azomethine ylide of gen .eral Formula (Ila): WO 98/50365 PCT/AU98/00312 -58- Y (Ha) wherein R 1 R 8 and R 14 may be the same or different and each are selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally protected hydroxy, optionally substituted amino, optionally substituted alkoxy, optionally substituted alkenoxy, optionally substituted alkynoxy, optionally substituted aryl, optionally substituted heterocyclyl, carboxy, carboxy ester, carboxamido, acyl, acyloxy, mercapto, optionally substituted alkylthio, halogen, nitro, sulfate, phosphate and cyano; and Y, Z and n are as defined in claim 1.

17. A method for the preparation of a compound of Formula (Ib) (Ib) comprising the step of cyclizing an azomethine ylide of general Formula (lib). -59- (OIb) r r wherein R 1 R 4 R 14 Y, Z and n are as defined in claim 16 and RA RA 4 form a cyclic-or non cyclic group as defined in claim 1.

18. A methodaccording to claim 17wherein RA RA 4 are hydrogen. A method according to claim 16'or claim 17, wherein Y is an optionally substituted phenyl group of Formula (ii): 20 wherein R 9 R' 3 are as defined for R' R 8 and R 14 in claim.16. A method according to claim 19, wherein R 9 -R 1 3 is selected from hydrogen, hydroxy, optionally substituted alkoxy, optionally substituted alkyl, sulfate and acyloxy.

21. A method according to claim 20, wherein R 9 -R 1 3 is selected from hydrogen, hydroxy, isopropoxy, methoxy, methyl, acetoxy and sulfate.

22. A method according to claim 16 or 17, wherein n is 0 or 1. WO 98/50365 PCT/AU98/00312

23. A method according to claim 22, wherein n is 1.

24. A method according to claim 16 or 17, wherein Z is carbon, nitrogen, sulfur or oxygen. A method according to claim 24, wherein Z is oxygen.

26. A method according to any one of claims 1, 16 or 17, wherein the cyclization of the azomethine ylide is achieved by thermal treatment.

27. A method according to claim 26, wherein the thermal treatment comprises heating the azomethine ylide in tetrahydrofuran, chloroform or 1,2- dichloroethane.

28. A method according to claim 27, wherein the thermal treatment further includes treatment with a metal salt.

29. A method according to claim 28, wherein the metal salt is Cul.

30. A method according to any one of claims 1, 16 or 17, wherein the cyclization is followed by oxidative treatment.

31. A method according to claim 30, wherein the oxidative treatment comprises oxidation in air.

32. A method according to claim 31, wherein the oxidative treatment in air is carried out in the presence of silica gel.

33. A method according to claim 30, wherein the oxidative treatment comprises treatment with Fremy's salt. WO 98/50365 PCT/AU98/00312 -61

34. A method according to claim 30, wherein the oxidative treatment comprises treatment with a quinone. A method according to claim 34, wherein the quinone is selected from chloranil or 2,3-dichloro-5,6-dicyano-l,4-benzoquinone.

36. A method according to claim 30, wherein the oxidative treatment comprises treatment with a metal catalyst. 0 37. A method according to claim 1, wherein the compound of Formula (II) is formed by treating a compound of Formula (III): (III) with a base, wherein A, Z, X, W, Y and n are as defined in claim 1 and the counter ion Le is a stable, weakly basic anion.

38. A method according to claim 16, wherein the compound of Formula (IIa) is formed by treating a compound of Formula (liIa):

62- Y (Ia) with a base, wherein R'-R 8 R 14 Y, Z and n are as defined in claim 13, and the counter ion LO is a stable basic anion. 39. A method according to claim 37 or claim 38, wherein L e is a sulphonate compound or a halogen. A method according to claim 39, wherein L e is selected from tosylate mesylate, triflate, bosylate, besylate, tresylate, nonaflate, nosylate, bromide, chloride or iodide. 41. A method according to claim 37 or claim 38, wherein the base is derived from an alkali metal, or the base is a mono-, di-, or tri-substituted amine. 42. A method according to claim 41, wherein the base is an alkyl lithium or an aryl lithium. 43. A method according to claim 41, wherein the base is an alkali metal carbonate. -63- 44. A mnethod according; to. claim 41 whejrei .n the. base is analicyl'.substJItuted amine. A method'according to claim 44, wherein the base is triethylamine or diisopropylethylammie. 46. A method according-to claim wherein the cyclization step is preceded by steps comprising: I O~A) coupling a: compound of Formula with a compound of Formula* Y (V) (Iv) to afford. the compound of Formula (VI): (VI) P4 wherein PZ,, is a synthon for b) unmasking Z. of compound (VI) and coupling with a compound L-CH 2 to provide compound (V11): L j I (VIII) -64- wherein L and L' are each a leaving group or a substituent convertible to a leaving group. c) treatment of compound (VII) with an imine of Formula (VIII) x S (vm W d) generation of the azomethine ylide of general formula (II); wherein Hal is halogen and A, W, Z, X, Y and n are defined in claims 37. 47. A method according to claim 16, wherein the cyclization step is preceded by steps comprising: coupling a compound of Formula (IV) to a compound of Formula (Va): y RS 6 H al R7 PZ, R 8 o o (Va) 20 to afford the compound of Formula (VIa): *R RS R 6 .o Y -R 7 (VIa) PZ R unmasking Z n of compound (Via) and coupling with a compound L-CH 2 L' to provide compound (VIIla): Rs~R6 -(R7 (Vwa) 1 Re L0 L treatment of compound (Vila) with a compound of Formula (Villa): R 4 R3 (VI a) R2 R14 R' generation of the azomethine ylide of general Formula (Ila); wherein Hal L, L', PZn, Y and Z are as defined in claim 46, and R'-R 8 and R 1 4 are as defined in claim 16. 48. A method according to claim 46 or claim 47, wherein PZ is OAc and unmasking refers to hydrolysis of the OAc group to provide OH. 49. A method according to claim 46 or claim 47, wherein PZ, is an aldehyde or acyl group and unmasking refers to oxidation of the aldehyde or acyl group to the corresponding ester, followed by hydrolysis. A method according to claim 46 ofrclaim 47, wherein L' is a halogen or OH converted to a leaving group. 51. A method according to claim 50, wherein L' is bromine or chlorine. Q:opcrtnjc\2239249 9 April.doc-O9It 12 -66- 52. A method according to claim 46 or claim 47, wherein the coupling step a) is carried out in the presence of a palladium catalyst. 53. A method according to claim 52, wherein the palladium catalyst is selected from PdCl 2 (PPh 3 2 or Pd(PPh 3 4 54. A method according to claim 52 or claim 53, wherein the coupling is mediated by a Cu(I) compound. A method according to claim 46 wherein one or more of the following steps: the coupling of a compound of Formula (IV) with a compound of Formula unmasking Zn of the compound of Formula (VI) and coupling with a compound L- CH 2 treatment of the compound of Formula (VII) with the imine of Formula (VIII), is performed by combinatorial synthesis. oooo oooo ooo o o ooo oo ooo oo ooo, oooo oooo o 56. A compound of Formula when prepared by a method according to claim 1, wherein A, Z, W, X, Y and n are as defined in claim 1. 57. A compound of Formula (Ia): WO 98/50365 PCT/AU98/00312 -67- when prepared by a method according to claim 16, wherein R' R 8 R 1 4 Y, Z and n, are as defined in claim 16. 58. A compound of Formula (Ib): when prepared by a method according to claim 17, wherein R 1 R 4 R 14 Y, Z, n and RAI RA 4 are as defined in claim 17. 59. A composition comprising a compound according to any one of claims 56 to 58, together with a pharmaceutically acceptable carrier, excipient or diluent. A compound of general Formula (I) Oe~jC239249 9 Apnil.doc-09AMM92 68 wherein A, Z, W, X, Y and n are as defined in claim 1, provided the compound is not one selected from Lamellarins A to N and S to X, Lamellarin I-acetate, Lamellarin B, C and G- diacetates, Lamellarin A, D, E, G, K, L, M and N-triacetates, Lamellarin H-hexaacetate, Lamellarin T, U, V and Y-20-sulphate, Lamellarin G-trimethyl ether and Lamellarin I- methylate. :0 .0*0 *0 0 0 61. A compound of general Formula (11) wherein A, Z, W, X, Y and n are as defined in claim 1. 62. A compound of general Formula (Ila):

69- Y t7a a a a a a 1' 4 wherein RJ-R 8 R 1 4 and Y, Z and n are defined in claim 16. 63. A compound: of general -Formula (l1b): (U1b) wherein R' R 4 R 14 Y, Z, n and RAI RA 4 are as defined in claim 17. 64. A compound. of general Formula (III). zI 0 X/aLe w (111) wherein A, Z, W, X, Y; n and LO are as defined in claim 37. 6. A compound of general Formula:(ila) (lila) '06* too* 06.90 wherin .R 1 R 8 Z.:ad L9 -ies defined in claim,38. 66. A compound of general: Formlula (flib)- (Illb) L 1 14 -IA40 wherein R -R4, R R Y, Z and n are as defined in claim 63 and L 6 is as defined in claim 64. -67. Use of a compound according to any one of claims 56 to 58, for the manufacture of a medicamnent for the treatment of multidrug resistant tumnours in a human or animal in need thereof. -71- 68. Use of a compound according to any one of claims 56 to 58, for the manufacture of a medicament for inducing apoptosis on a multidrug resistant cell in a human or animal in need thereof. 69. Use of a compound according toany one of claims56 to 58, for the manufacture of a medicament for improving the anti-tumour chemotherapeutic effect on multidrug resistant affected drugs in a human or animal in need thereof. Use of a compound according to any one of claims 56 to 58, for the manufacture of a medicament for modulating immunological functions in a human or animalin need thereof. the administration of an effective amount of a compound according to any one of

72. A method for inducing a tosis of multidrugresistant cells in a teadministration of an ffective amou f compound according t one Shuman or animal, comprising the administration of an effective amount of a compound Saccording to any one of claims 56 to 58, to a human or anmal in need thereof. %woo:: oo•

73. A method for improving the anti-tumour chemotherapeutic effect on multi-drug resistant cells comprising the administration of an effective amount of a compound according to any one of claims 56 to 58, to a human or animal in need thereof.

74. A method for modulating immunological functions comprising the administration of an effective amount of a compound according to any one of claims to 57, to a human or animal in need thereof. PAOPERMCM798 spCAMc22/4/02 -71A- A method according to claim 52 wherein the coupling is mediated by a Cu(I) compound and wherein the palladium catalyst is selected from PdCl 2 (PPh 3 2 or Pd(PPh 3 4 DATED this 22 nd day of April, 2002 The Australian National University 10 By DAVIES COLLISON CAVE Patent Attorneys for the Applicant