Pharmaceutically active normonyl compounds.

This invention relates to a novel class of compounds having antibacterial and antimycoplasmal activity, to processes for their preparation and to their use in human and veterinary medicine, and also to intermediates for use in the preparation of such compounds.

Mupirocin, the compound of formula (A) :

exhibits good activity against Gram positive bacteria, H.influenzae, Leαionella and mycoplasma. It is marketed as a topical formulation by Beecham Group p. I.e. under the Trade Mark 'Bactroban' . Mupirocin is however rapidly hydrolysed _in vivo to monic acid, the compound of formula (B) :

(B)

which is inactive.

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Various proposals have been made to improve the metabolic stability of mupirocin with respect to enzymatic hydrolysis. Thus, in the past, the impact of various modifications to the ester functional group have been investigated, for instance, replacing this functional group by a heterocyclic derivative (EP-A-0 087 953 and EP-A-0 123 578, Beecham Group p.I.e.), an amide (US 4 312 764, Beecham Group Ltd, EP-O-029 665) or a ketone, in particular an alkyl ketone (Coulton e_t al in J. Chem. Soc Perkin I, 1982, 729) . More recently, the phenyl ketone has been reported by Klein e_t al. (in a poster presented at the Third Annual Chemical Congress of North America, Toronto, June 1988 and followed up in J. Med. Chem. 1989, 32., 151) . The activity of this derivative (as judged by MIC values against a range of S. aureus bacterial strains) was however reduced in comparison to mupirocin. Furthermore, when the series was extended to include substituted phenyl ketones, it was found that although a 4-fluorine substituent had little e ^' ffect on activity, further fluorine substitution reduced the activity, whilst 2,4, 6-trimethyl substitution drastically reduced activity. No information was however provided concerning in vivo stability.

Surprisingly, it has now been found that with other patterns of phenyl substitution, an enhanced overall biological profile may be observed.

Accordingly, the present invention provides a compound of formula (I) :

B,C

in which R° denotes a substituted phenyl group; and excluding the compounds 4-fluorophenyl-l-normon-2-yl ketone, 2, 4 , 6-trifluorophenyl-l-normon-2-yl ketone, 2, 3, ,5, 6-pentafluorophenyl-l-normon-2-yl ketone and 2, 4, 6-trimethylphenyl-l-normon-2-yl ketone.

Suitable substituents for the phenyl group of R° include, for example, hydroxy; ercapto; ureido; amino; halogen; cyano; nitro; formyl; a group of the formulae -X^-R ,-Y^-R ⁶ , -X ¹ -Y ¹ -R ^β , -Y ¹ -X ¹ -R ^β , -X ¹ -Y ¹ -X ² -R ⁶ or R ^β , in which X ¹ and X , which may be the same or different, is each selected from 0, S, NH, or NR ⁷ , Y ¹ is CO or S0 , and R ^β and R ⁷ , which may be the same or different, is each selected from (C-^- c ₎ ) a.lkγl , (C3-7) cycloalkyl, (C2-9) alkenyl, (C3~g) cycloalkenyl, and (C2- ) alkynyl, each of which may be optionally substituted, aryl, or heterocyclyl; a group

-(SO)R , in which R° is as hereinbefore defined; a group _ _y 2 _R 9 _R 10 _{in wnich y} 2 _is ^ _{NC0/ or NSQ2 and R} 9 _{and R} 10 _form an optionally substituted (C3-7) alkylene chain optionally interrupted by 0,S or NR ⁶ , in which R° is as hereinbefore defined; a group -C(X ³ )H or -C(X ³ )R ⁶ in which X ³ is =N0H, =NOR ⁷ or =NNR ^β R ⁷ and R ^β and R ⁷ are as hereinbefore defined; or a divalent group Y ^J which, together with the two carbon atoms of the phenyl group to which it is bonded, forms a heterocyclic or carbocylic ring.

The phenyl group of R° may have up to five substituents, suitably up to three substituents, preferably up to two substituents, and advantageously one substituent.

Preferably in the group R°, no more than one of the substituents of the phenyl group will be in an ortho disposition relative to the carbonyl group of the ketone. Suitably a substituent is located para to the carbonyl group of the ketone.

Preferably, if one of the substituents is fluoro, then the phenyl group of R° has at least one other substituent which is not fluoro.

Suitably a substituent for the phenyl group of R° is selected from: halogen; hydroxy; amino; ureido; cyano; formyl; optionally substituted (C- _j __g) alkyl; ( 2_g) alkenyl; (C ₂ _g) alkynyl; heteroaryl; a group -OR ⁶ , -SR ⁶ , -NHR ⁶ or -NR ⁶ R ⁷ , -NHCOR ⁶ , -COR ⁶ , -(SO)R ⁶ , -S0 ₂ R ^β , -C(=NOH)H,

-(C=NOR ⁶ )H, -C(=NOH)R ⁶ , -C(=NOR ^β )R ⁷ , -C(=NNR ⁶ R ⁷ ) in which R and R are as hereinbefore defined and are each preferably optionall in which

Preferably, the divalent group Y ³ is bonded to two carbon atoms which are in a 1,2-relationship.

Typically the divalent group Y ³ may be formed by the remova of a hydrogen atom from a monovalent substituent for the phenyl group of R°. Alternatively, Y ³ may be formed from two monovalent substituents for the phenyl group of R°, which may be the same or different, linked by the removal o a hydrogen atom from each.

Examples of a divalent group Y ³ include (^-3)alkylenedioxy, for instance methylenedioxy, and a -(CH=CH)NH- group, such that R° is indolyl.

When used herein, the term 'aryl' includes, unless otherwise defined, phenyl or naphthyl optionally substituted with up to five, preferably up to three substituents.

When used herein, the term 'heterocylyl' or 'heterocyclic' includes single or fused aromatic or non-aromatic rings comprising up to four hetero-atoms in the ring selected from oxygen, nitrogen and sulphur and optionally substituted with up to three substituents. Suitably the heterocyclic ring comprises from 4 to 7, preferably 5 to 6 , ring atoms.

When used herein, the term 'heteroaryl' includes an aromatic heterocyclic ring or ring system, preferably with 5 or 6 ring atoms on each ring.

When used herein, the term 'halogen' refers to fluorine, chlorine, bromine or iodine.

Substituents for groups hereinbefore defined as being optionally substituted, for instance alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl or heterocyclyl, include:

(a) halogen, cyano, azido, nitro, phthalimido, formyl, carboxy, carboxylate salts, sulpho, sulphonate salts, or oxo;

(b) amino, imino, hydrazino, hydrazono, ureido, guanidino, carbamoyl, or sulphonamido, in each of which groups a nitrogen may be further optionally substituted by one or two groups (which may be the same or different) selected from the groups listed in subparagraphs (d) , (e) and (f) ;

(c) hydroxy, oxyimino, hydroximoyl, benzohydroximoyl, or mercapto, in each of which groups hydrogen may be replaced by one of the groups listed in subparagraphs (d), (e) and (f) ;

(d) a group RP wherein RP denotes aryl or heterocyclyl;

(e) a group R^ wherein R ^q denotes (C-^-g) alkyl,

(C3--7) cycloalkyl, (C2-g) lkenyl, (C3-3) cycloalkenyl, or ( 2-g) alkynyl, each of which may be optionally substituted by up to three groups (which may be the same or different) chosen from the groups listed in subparagraphs (a) , (b) , (c) , (d) and (f) ; and

(f) a group RPCO-, RPOCO-, R ^q C0-, R^OCO-, RPSO-, RPS0 ₂ -, R ^q SO-, and R ^q S0 ₂ - wherein R ^p and R ^q are as defined in subparagraphs (d) and (e) respectively.

Suitable substituents for an alkyl, cycloalkyl, alkenyl, cycloalkenyl or alkynyl group include for example, halogen, cyano, azido, nitro, carboxy, (C-^- ) alkoxycarbonyl, carbamoyl, mono- or di- (C-^-g) alkylcarbamoyl, sulpho, sulphamoyl, mono- or di- (C _] _-g) alkylsulphamoyl, amino, mono- or di- (C- _j _-g) alkylamino, acylamino, ureido, (C-^-g) alkoxycarbonylamino, 2, 2, 2-trichloroethoxycarbonyl- amino, aryl, heterocyclyl, hydroxy, (C-^-g) alkoxy, acyloxy, oxo, acyl, 2-thenoyl, (C-^-g) alkylthio, (C-^-g) alkylsulphinyl, (C- _j _-g) alkylsulphonyl, hydroxyimino, (C- _| _-g) alkoxyimino, hydrazino, hydrazono, benzohydroximoyl, guanidino, amidino, or iminoalkylamino.

Suitable substituents for an aryl group include, for example, halogen, cyano, (C- _| _-g) alkyl, phenyl, (C-^-g)alkoxy, halo (C- _| _-g) alkyl, hydroxy, amino, mono- or di- (C-^-g) alkylamino, acylamino, nitro, carboxy,

(C ₁ -g)alkoxycarbonyl, (C ₁ -g)alkoxycarbonyl (C ₁ ~g)alkyl, (C^-g) alkylcarbonyloxy, (C^- ) alkylthio, (Cη _^ - )alkylsulphinyl, (C ₁ ~g) alkylsulphonyl, sulphamoyl, mono- or di- (C^-g) alkylsulpha oyl, carbamoyl, and mono- or di-(C^-g) alkylcarbamoyl.

Suitable substituents for a heterocyclyl group include, for example, halogen, (C-^-g)alkyl, (C- _j^ -g) alkoxy, halo ( ;L_g) alkyl, hydroxy, amino, mono- or di- (C _] _-g) alkylamino, carboxy, (C-^-g) alkoxycarbonyl, (C ₁ -g) alkoxycarbonyl (C^-g) alkyl, aryl or oxo.

Compounds of formula (I) may conveniently be named ' (l-normon-2-yl)ketones' . Normonyl is the trivial name for the 3-[ (2S,3R,4R,5S)-5-[ (2S, 3S, S, 5S) -2, 3- epoxy-5-hydroxy-4-methylhexyl]-3, 4-dihydroxytetrahydro- ρyran-2-yl]-2-methylprop-l (E) -enyl radical, as shown in formula (II) :

It will be appreciated that in compounds of formula (I) the absolute and relative stereochemistry of each of the chiral centres in the normonyl moiety thereof will be the same as that in the corresponding moiety of monic acid A.

Furthermore, it will also appreciated that in compounds of formula (I), the group R° may contain one or more chiral centres. The present invention encompasses all such resultant isomeric possibilities.

Since the compounds of formula (I) of the present invention are intended for use in pharmaceutical compositions, it will be understood that they are each provided in substantially pure form, for example at least 50% pure, more suitably at least 75% pure and preferably at least 95% pure (% are on a wt/wt basis) . Impure preparations of the compounds of formula (I) may be used for preparing the more pure forms used in the pharmaceutical compositions. Although the purity of intermediate compounds of the present invention is less critical, it will be readily understood that the substantially pure form is preferred as for the compounds of formula (I) . Preferably, whenever possible, the compounds of the present invention are obtained in crystalline form.

When some of the compounds of this invention are allowed to crystallise, or are recrystallised, from organic solvents. solvent of crystallisation may be present in the crystalline product. This invention includes within its scope such solvates. Similarly, some of the compounds of this invention may be crystallised or recrystallised from solvents containing water. In such cases water of hydration may be formed. This invention includes within its scope stoichiometric hydrates as well as compounds containing variable amounts of water that may be produced by processes such as lyophilisation.

Examples of compounds within this invention include the following:

p-Methoxyphenyl-l-normon-2-yl ketone;

m-Methoxyphenyl-l-normon-2-yl ketone;

3, 4-Methylenedioxyphenyl-l-normon-2-yl ketone;

l-Normon-2-yl-ρ-methylthioρhenyl ketone;

- 9- l-Normon-2-yl-p-methylsulphinyl ke one;

p-Hydroxyρhenyl-l-normon-2-yl ketone;

ρ-Dimethylaminophenyl-l-normon-2-yl ketone;

ρ-Methylsulρhonyl-l-normon-2-yl ketone;

4-Cyanophenyl-l-normon-2-yl ketone;

3-Fluoro-p-methoxyphenyl-l-normon-2-yl ketone;

2-Methoxyphenyl-l-normon-2-yl ketone;

4-Hydroxymethylphenyl-l-normon-2-yl ketone;

4-Formylphenyl-l-normon-2-yl ketone;

4-Acetylphenyl-l-normon-2-yl ketone;

4- (2-Hydroxyethoxyiminomethyl)phenyl-l-normon-2-yl ketone;

4- (4-Hydroxybutoxyiminomethyl)phenyl-l-normon-2-yl ketone;

4- (l-Hydroxyethyl)phenyl-l-normon-2-yl ketone;

l-Normon-2-yl-4- (ρyrrolidin-1-ylsulphonyl)phenyl ketone;

4- (2-Hydroxyethoxy)ρhenyl-l-normon-2-yl ketone;

4-Methylthiomethoxyphenyl-l-normon-2-yl ketone;

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4- (1-Hydroxyimino)ethylρhenyl-l-normon-2-yl ketone;

5-Indolyl-l-normon-2-yl ketone;

52-Hydroxyρhenyl-l-normon-2-yl ketone;

4-Hydroxyiminomethylphenyl-l-normon-2-yl ketone;

4-Methoxyiminomethylphenyl-l-normon-2-yl ketone; 10 l-Normon-2-yl 4-propylthioρhenyl ketone;

4-Propylsulphinylphenyl-l-normon-2-yl ketone;

154-Propylsulphonyϊphenyl-l-normon-2-yl ketone;

4-Methylsulphinylmethoxyρhenyl-l-normon-2-yl ketone;

4-Methylsulphonylmethoxyphenyl-l-normon-2-yl ketone; 20 p-Aminophenyl-l-normon-2-yl ketone;

l-Normon-2-yl-p-ureidophenyl ketone;

25 p-Ethylaminophenyl-l-normon-2-yl ketone;

l-Norm-2-yl-p-proρanamidophenyl ketone;

p-Imidazol-4 (5)-yl phenyl-l-normon-2-yl ketone; 30 l-Normon-2-yl-ρ-pyrimidin-4-yl phenyl ketone;

p-Ethynylphenyl l-normon-2-yl ketone;

l-Normon-2-yl p-trifluoromethylphenyl ketone;

p-Ethenylphenyl l-normon-2-yl ketone;

p-l-Methylimidazol-4-yl phenyl-l-normon-2-yl ketone;

p-1-Methyl imidazol-5-yl phenyl-l-normon-2-yl ketone; and

p-Ethylphenyl l-normon-2-yl ketone.

Compounds of the present invention may be prepared by methods known for the preparation of α,β-saturated ketones Some of these processes will be more appropriate than others.

Suitably, compounds of formula (I) may be prepared by a process which comprises treating a compound of formula (III) :

(III)

in which Z 1 , Z? and Z ^~ "" are the same or different and each is hydrogen or a hydroxyl-protecting group.

or a derivative thereof in which the carboxyl group is activated.

with an organometallic reagent;

and thereafter, and if necessary, removing any hydroxyl-protecting groups.

Suitable organometallic reagents include:

(i) a Grignard reagent of the formula R°MgX ⁴ in which R° is as defined with respect to formula (I) and X ⁴ represents chlorine, bromine or iodine, which reaction may optionally be carried out in the presence of copper(I) iodide as catalyst;

(ii) an organolithium reagent of formula R°Li in which R° is as defined with respect to formula (I) ;

(iii) an organomanganous reagent of the formula R°MnCl in which R° is as defined with respect to formula (I) ; and

(iv) an organocerium reagent R°Li-Ce(X ⁴ ) 3, in which R° is as defined with respect to formula (I) and X ⁴ represents chlorine, bromine or iodine.

The reaction with the organometallic reagent may be conveniently carried out in an ethereal or hydrocarbon solvent, the choice of which is dependent upon the specific requirements of the organometallic reagent. Preferably, the Grignard reagent is generated and used in diethyl ether or tetrahydrofuran.

The reaction is generally carried out in an inert atmosphere such as argon or nitrogen and at ambient temperature or below. The period for which the reaction is allowed to

proceed depends upon the particular starting materials employed. The course of the reaction may be followed by conventional methods such as thin layer chfomatography and the reaction may be terminated when an optimum quantity of product is present in the reaction mixture.

Suitable derivatives of the compound of formula (III) in which the carboxyl group is activated include for instance thio-esters of formula (IV) :

1 -3 in which Z , Z^ and Z ^J are as hereinbefore defined and the moiety: „ - --»

represents a 5- or 6-membered heterocyclic ring which may contain in addition to the nitrogen atom, one or two further heteroatoms selected from oxygen, nitrogen and sulphur and which may be substituted or fused to a benzene ring which may itself be substituted.

Preferred thio-esters are of formula (IVa)

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in which Z , Z ² and Z ³ are as hereinbefore defined,

A compound of formula (IVa) may be prepared by the treatment of a compound of formula (III) with 2,2'-dipyridyl disulphide in the presence of triphenylphosphine, by analogy with the method described by E.J. Corey and D.A Clark in Tetrahedron Lett, 1979, 31, 2875.

Other suitable derivatives of the compound of formula (III) include mixed anhydrides of the formula (V) :

in which Z , Z , and Z ^J are as hereinbefore defined. and

R ¹¹ is (C-L-g) alkyl;

and of the formula (VI)

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in which Z , Z , and Z ³ are as hereinbefore defined, and R ¹² and R ^J are the same or different and each denotes an aryl group, for instance phenyl, or a (C-^-gJalkoxy group, for instance ethoxy.

Further suitable derivatives of the compound of formula (III) include amides of the formula (VII) :

in which Z , Z ² and Z ³ are as hereinbefore defined, R ¹⁴ and R .1 ¹ 5 are the same or different, and each is ( _] _~ ) alkyl, or the substituents R ,14 and R ,15 ^o form a (C ₂ - ₇ ) alkylene chain;

and of the formula (VIII)

-16-

in which Z , Z ² , and Z ³ are as hereinbefore defined and R ¹⁶ 10 and R , together with the nitrogen atom to which they are bonded, form an imidazolyl or triazolyl ring.

Preferred amides of formula (VII) and (VIII) include the N-methoxy-N-methyl and the imidazol-1-yl derivatives 15 respectively.

The reaction of an N-methoxy-N-methylamide with an organolithium or a Grignard reagent to form a ketone is described by Nahm and Weinreb in Tetrahedron Lett, 1981, 203815. The reaction of an α,β-unsaturated acid or its imidazolyl derivative with a Grignard reagent is described in Chem. Ber., 1965, .95 1284.

Amides of formula (VII) and (VIII) may be obtained from a 25 compound of formula (III) by conventional methodology.

Suitably a compound of formula (III) is converted to a mixe anhydride of formula (V) in which R is iso-butyl, by treating the compound of formula (III) with iso- butylchloroformate in the presence of triethylamine, in 30 tetrahydrofuran at from -5 to +20°C. The mixed anhydride thus obtained may then be treated with either an amine HN(OR ¹⁴ )R ¹⁵ or an amine hydrochloride HNR ¹⁶ R ¹⁷ .HC1, in whic R ¹⁴ to ^x7 are as hereinbefore defined, to give an amide of formula (VII) or (VIII), respectively. 35

Suitably a thio-ester of formula (IV) is treated with an organomanganous reagent of formula R°MnCl, as hereinbefore defined.

Suitably an amide of formula (VII) or (VIII) is treated with an organolithium reagent of formula R°Li or an organocerium reagent of formula R°Li-Ce (X ) 3 as hereinbefore defined.

Suitable organometallic reagents may be prepared according to conventional procedures.

Suitable organomanganous reagents of the formula R°MnCl may be conveniently prepared by addition of an organolithium reagent R°Li to a solution of manganous chloride and lithium chloride in dry THF, or a suspension of anhydrous manganous chloride in dry THF. An excess of R°MnCl is preferably employed. Alternatively, a Grignard reagent may be used in place of the organolithium reagent, to generate the organomanganous reagent R°MnCl.

Other organomanganous reagents which may be used instead of R°MnCl include:

(i) (R°) ₃ MnLi or (R°) ₃ MnMgX ⁴ in which X ⁴ is as hereinbefore defined, as described in Synthetic Communications, 1979, 9_, 639;

(ii) R°MnI in ether; as described in Synthetic Communications, 1979, I_, 639; and

(iii) R°MnBr in ether, as described in Tetrahedron Lett, 1976, 3155.

As in the case of R°MnCl, the above organomanganous reagents may be prepared .in. situ when required.

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Organoceriu reagents may be generated in situ by treatment of an organolithium compound of the formula R°Li, in which R° is as hereinbefore defined, with cerium (III) halide, by analogy with the procedure described by Imamoto et al; J.Chem. Soc. , Chem. Commun, 1982, 1042.

Other processes for the preparation of compounds of formula (I) are described in EP-A-0 029 665 (Beecham Group Ltd) and include treating an allylic alcohol of formula (IX) :

in which R°, Z , Z and Z ³ are as hereinbefore defined, with an oxidising agent which converts allylic alcohols into α,β-unsaturated ketones, and thereafter, and if necessary, removing any hydroxyl-protecting groups.

Suitable such oxidising agents include activated manganese dioxide, pyridinium dichromate and pyridinium chlorochromate.

Conveniently, the oxidation reaction is carried out in a polar organic solvent.

An allylic alcohol of formula (IX) may be prepared by treating the corresponding aldehyde of formula (X) :

(X)

in which Z ¹ , Z ² and Z ³ are as hereinbefore defined,

with an organometallic reagent as hereinbefore defined, and thereafter, and if necessary, removing any hydroxyl-protecting groups.

Suitably, an aldehyde of formula (X) is treated with a

Grignard reagent of formula R°MgX , as hereinbefore defined.

An aldehyde of formula (X) may be prepared by analogy with the methodology described in EP 0 029 665 (Beecham Group Ltd.) .

A compound of formula (I) may also be prepared by treating a ketone of formula (XI) :

1 ? "^ n which Z , Z^ and Z ^J are as hereinbefore defined, with a terminal alkyne of the formula (XII) :

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(XII) in which R° is as hereinbefore defined.

to form an intermediate which is treated with tris (triphenylsilyloxy)vanadate and triphenylsilanol, as described by H. Pauling in Helvetica, 1976, 5_9, 1233 and G.L. Olson, Helvetica, 1976, 5_9 567;

10 and thereafter, and if necessary, removing any hydroxyl-protecting groups.

The present invention further provides a process for 15 preparing a compound of formula (I) which process comprises subjecting a compound of formula (XIII) :

25 in which R°, Z ¹ , Z ² and Z ³ are as hereinbefore defined, to isomerising conditions which will convert the Z-double bond of the compound of formula (XIII) to an E-double bond.

Suitable isomerising conditions are described by Sonnet in 30 Tetrahedron, 1980, 3_6, 557 and include photo-chemical and addition-elimination methods.

A compound of formula (XIII) may be prepared by treating a compound of formula (XI) with a compound of formula (XII) , as hereinbefore described. This reaction is lacking stereoselectively and may lead to the formation of compounds of formulae (I) and (XIII),which may then be separated by conventional procedures such as chromatography.

Compounds of formula (I) may be obtained from other" compounds of formula (I) by suitable manipulation of the substituents present in the group R° according to conventional methodology. For instance, an alkylthio substituent may be converted to an alkyl sulphinyl or an alkylsulphonyl substituent by treatment thereof with a conventional oxidising agent such as m-chloroperbenzoic acid. Also included within this is salification and esterification of a carboxy substituent, trans- and de-esterification of an ester containing substituent, reduction of an alkoxycarbonyl substituent, formation of the free carboxy group from a carboxylate salt, formation of an oxime derivative from the corresponding carbonyl compound, acylation of an amine substituent and oxidation of a hydroxyl group.

When used herein, the term 'hydroxyl-protecting group' refers to any such group known in the art which may be removed without disruption of the remainder of the molecule.

Suitable hydroxyl-protecting groups are described in

'Protective Groups in Organic Synthesis', T.W. Greene,

Wiley-Interscience, New York 1981.

The hydroxyl groups of the compounds of formulae (III) to

(XI) and (XIII) may be protected at any stage of the above processes, using conventional methods. The hydroxyl-protecting group may be removed by methods known in the art, including enzymatic methods.

-22-

Particularly suitable hydroxyl-protecting groups are silyl groups since these are readily removed under mild conditions. Such groups are introduced using conventional silylating agents, including halosilanes and silazanes, of the formulae below:

L ₃ SiX° L SiO-C=NSiL- L SiX L L ₃ SiNL ₂ L ₃ SiNHSiL ₃ Me ₃ Si-N N L ₃ SiNHCOL L ₃ SiNHCONHSiL ₃ -BuMe ₂ Si- -N N LNHCONHSiL-3

wherein Me denotes methyl and ^t Bu denotes t-butyl, X ⁵ is halogen and each group L is independently selected from hydrogen, (C^g)alkyl, (C^-g)alkoxy, aryl or aryl (C- _j^ -^) alkyl. A preferred silyating agent is trimethylsilyl chloride. Particularly suitable protecting groups are trimethylsilyl, t-butyldimethylsilyl and t-butyldiphenylsilyl groups. Preferred protecting groups are trimethylsilyl groups because of their ease of removal

The glycol function of the compounds of formulae (III) to (XI) and (XIII) may be protected by forming a cyclic derivative using a compound of formula (XIV) :

R ¹⁸ - C - OR ²⁰ (XIV)

OR 21

wherein R 1 ^x 8 ^o is hhyrdlrmongaen a al "1k lryl 1 a ann Hd oeaachh rosf Rl °, 2 0 aanndd RR ²²¹¹ iiss ((CC- ₁ ^-ggJ)aallkyyl. In the cyclic derivative Z ² and Z ³ together are a moiety

/

wherein R ²² is (C-^-g) alkyl.

Suitably R ¹⁸ is hydrogen, methyl, ethyl, n- or iso-propyl; most suitably it is hydrogen. The groups R , R ²⁰ and R ²¹ are suitably methyl, ethyl, n- or iso-propyl, or n-, iso-, sec- or t-butyl; most suitably methyl.

Similarly the hydroxyl groups of a compound of formula (I) may be protected prior to conversion to a further compound of formula (I) as described above.

In each case the hydroxyl-protecting groups described above may be removed by mild acid hydrolysis followed by alkaline hydrolysis, for instance, as described by J.P. Clayton, K. Luk and N.H. Rogers, in 'Chemistry of Pseudomonic Acid, Part II', J.C.S. Perkin Trans. 1 , 1979, 308.

The compounds of this invention are useful for the treatment of bacterial and mycoplasma-induced infections in non-human and human animals, such as the treatment of respiratory tract infections, otitis, meningitis, skin and soft tissue infections in human animals, mastitis in cattle, and respiratory infections in non-human animals such as pigs and cattle.

The compounds of this invention are active against both Gram negative and Gram positive organisms, including Haemophilus, for instance H.influenzae Ql; Branha ella,for instance B.Catarrhalis 1502; Streptococci, for instance S.pyoqenes CN10 and S.pneumonia PU7; and Staphylococci, for instance S.aureus Oxford, and against mycoplasma. In addition, compounds of the present invention are active against Staphylococci organisms such as S.aureus and S.epidermis which are resistant, including multiply resistant, to other anti-bacterial agents, for instance, macrolides, aminoglycosides, lincosamides and β-lactams, such as for example, methicillin.

This invention also provides a pharmaceutical or veterinary composition which comprises a compound of formula (I) (hereinafter referred to as the 'drug') together with a pharmaceutically or veterinarily acceptable carrier or excipient.

The compositions may be formulated for administration by any route, and would depend on the disease being treated. The compositions may be in the form of tablets, capsules, powders, granules, lozenges, liquid or gel preparations, such as oral, topical or sterile parenteral suspensions.

Tablets and capsules for oral administration may be in unit dose presentation form, and may contain conventional excipients such as binding agents, for example syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinylpyrollidone; fillers, for example lactose, sugar, maize-starch, calcium phosphate, sorbitol or glycine; tabletting lubricants, for example magnesium stearate, talc, polyethylene glycol or silica; disintegrants, for example potato starch, or acceptable wetting agents such as sodium lauryl sulphate. The tablets may be coated according to methods well known in normal pharmaceutical practice

Oral liquid preparations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be presented as a dry product for reconstitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents, for example sorbitol, syrup, methyl cellulose, glucose syrup, gelatin, hydrogenated edible fats; emulsifying agents, for example lecithin, sorbitan monooleate, or acacia; non-aqueous vehicles (which may include edible oils) , for example almond oil, fractionated coconut oil, oily esters, glycerine, propylene glycol, or ethyl alcohol; preservatives, for example methyl or propyl p_-hydroxybenzoate or sorbic acid, and if desired conventional flavouring or colouring agents.

For topical application to the skin the drug may be made up into a cream, lotion or ointment. Cream or ointment formulations that may be used for the drug are conventional formulations well known in the art, for example, as described in standard text books of pharmaceutics and cosmetics, such as 'Harry's Cosmeticology' published by Longman, and the British Pharmacopoeia.

Suppositories will contain conventional suppository bases, e.g. cocoa-butters or other glyceride.

For parenteral administration, fluid unit dosage forms are prepared utilizing the drug and a sterile vehicle. The drug, depending on the vehicle and concentration used, can be suspended in the vehicle. Advantageously, adjuvants such as a local anaesthetic, preservative and buffering agents can be dissolved in the vehicle. To enhance the stability the composition can be frozen after filling into the vial and water removed under vacuum. The dry lypophilized powder is then sealed in the vial. The drug can be sterilised by

exposure to ethylene oxide before suspending in the sterile vehicle. Advantageously, a surfactant or wetting agent is included in the composition to facilitate uniform distribution of the drug.

For topical application to the ear, the drug may be made up into a suspension in a suitable liquid carrier, such as water, glycerol, diluted ethanol, propylene glycol, polyethylene glycol or fixed oils.

For topical application to the eye, the drug is formulated as a suspension in a suitable, sterile aqueous or non-aqueous vehicle. Additives, for instance buffers such as sodium metabisulphite or disodium edetate; preservatives including bactericidal and fungicidal agents, such as phenylmercurie acetate or nitrate, benzalkonium chloride or chlorhexidine, and thickening agents such as hypromellose may also be included.

The dosage employed for compositions administered topically will, of course, depend on the size of the area being treated. For the ears and eyes each dose will typically be in the range from 10 to 100 mg of the drug.

Veterinary compositions for intramammary treatment of mammary disorders in animals, especially bovine mastitis, will generally contain a suspension of the drug in an oily vehicle.

The compositions may contain from 0.1% to 99% by weight, preferably from 10-60% by weight, of the drug,depending on the method of administration. Where the compositions are in unit dose form, each dosage unit will preferably contain from 50-500 mg, of the drug. The dosage as employed for

adult human treatment (average weight about 70 kg) will preferably range from 100 mg to 3 g, per day, for instance 250 mg to 2 g of the drug per day, depending on the route and frequency of administration.

Alternatively, the drug may be administered to non-human animals as part of the total dietary intake. In this case the amount of drug employed may be less than 1% by weight of the diet and in preferably no more than 0.5% by weight. The diet for animals may consist of normal foodstuffs to which the drug may be added or the drug may be included in a premix for admixture with the foodstuff. A suitable method of administration of the drug to a non-human animal is to add it to the non-human animal's drinking water. In this case a concentration of the drug in the drinking water of about 5-500 μg/ml, for example 5-200 μg/ml, is suitable.

The present invention further provides a method for treating the human or non-human animal which method comprises administering a therapeutically effective amount of a compound of formula (I) as hereinbefore defined, to a human or non-human animal in need of such therapy.

The present invention also provides a compound of formula (I) as hereinbefore defined for use in therapy.

Alternatively, a pharmaceutical composition as hereinbefore described may be employed in the treatment.

In particular aspects of the treatment there are provided methods for treating bacterial infections and/or mycoplasma induced infection of human or non-human animals, especially respiratory infections in human or non-human animals.

The present invention further provides for the use of a compound of formula (I) as hereinbefore defined in the manufacture of a medicament for use in anti-bacterial or anti-mycoplasmal therapy.

The following Examples illustrate the invention, but are not intended to limit the scope in any way.

Example 1

p-Methoxyρhenyl-l-normon-2-yl ketone

5 a) 6,7, 13-0-Tris-trimethylsilylmonic acid

Monic acid A (1.72g, 5mmol) was dissolved in dry tetrahydrofuran (50ml) and triethylamine (4.7ml, 30mmol) . Chlorotrimethylsilane (3.7, 30mmol) and 10 4-dimethylaminopyridine (20mg) were added, and the mixture stirred at 20°C for 3h.

The reaction mixture was filtered, the solid washed with dry tetrahydrofuran, and the combined filtrates concentrated

15 under reduced pressure. The residue was taken up in diethyl ether and washed with water and saturated brine. After drying (MgS0 ₄ ) the solvent was removed by evaporation under reduced pressure to yield 6 , 1 , 13-O-tris-trimethvlsilylmonic acid (2.24g, 83%); V _maχ (CH ₂ C1 ₂ ) 3300-2400, 2960, 2900,

20 1685, 1640cm- ¹ .

b) S-2-Pyridyl 6, 7, 13-O-Tris-trimethylsilylmonthioate

To triphenylphosphine (1.05g, 4mmol) in dichloromethane 25 (1ml) was added di-2-pyridyldisulphide (0.88g, 4mmol) . The mixture was cooled to 0°C and a solution of the above tris-protected monic acid (1.08g, 2mmol) in dichloromethane (4ml) added. The reaction was stirred at 20°C for 6h, then triethylamine (0.15ml) added and the crude mixture purified 0 by chromatography on silica gel eluting with a gradient of ethyl acetate in hexaneto give S-2-pyridyl 6, 7, 13-O-tris-trimethylsilylmonthioate as a yellow gum (0.804g, 61%); v _maχ (film) 3040, 2960, 2900, 1690, 1620, 1570, 1560, 1250cm- ¹ ; δ _R (CDCI3) (inter alia) 0.1-0.2 (27H, 5 m, SiMe) , 0.9 (3H, d, J 7.0Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.3Hz,

-30-

14-H ₃ ), 2.05 (1H, dd, J 10.5, 14.6Hz, 4-H) , 2.2 (3H, s, I5-H3), 2.55 (1H, d, J 14.6Hz, 4-H), 2.7 (2H, m, 10 and 11-H), 6.15 (1H, s, 2-H) , 7.3 (1H, m, pyridine 5-H) , 7.65 (1H, d, J 7.9Hz, pyridine 3-H) , 7.75 (1H, m, pyridine 4-H), 8.65 (1H, m, pyridine 6-H) .

c) ρ-Methoxyphenyl-l-nσrmon-2-yl ketone

To p_-methoxyphenyl magnesium bromide [prepared from p_-bromoanisole (0.5ml, 4mmol) and magnesium turnings

(0.097g, 4mmol) in tetrahydrofuran (5ml)] cooled to -15°C was added manganese (II) chloride (0.504g, 4mmol) . The mixture was allowed to warm to 20°C and stirred at this temperature for lh. After cooling to -15°C a solution of the thiol ester from part (a) (1.31g, 2mmol) in tetrahydrofuran (5ml) was added dropwise. The reaction mixture was allowed to warm to 20°C and stirred at this temperature for 90min. Saturated aqueous ammonium chloride (30ml) was added, and the mixture extracted with ethyl acetate. The organic layer was dried (MgS0 ₄ ) , concentrated by evaporation under reduced pressure, and partially purified by chromatography on silica gel eluting with diethyl ether in hexane (0-20%) , to give impure tris-protected product (1.53g) .

This material was dissolved in tetrahydrofuran (35ml) and 0.4M hydrochloric acid (9ml) added. After 3min. at 20°C the reaction was quenched with saturated aqueous sodium hydrogen carbonate, and extracted with ethyl acetate. After drying (MgS0 ₄ ) and concentration under reduced pressure, the residue was purified by chromatography on silica gel eluring with methanol in dichloromethane (0-6%) to give the title product as a colourless foam, (0.247g, 28%); v _maχ (KBr) 3427, 2963, 1649, 1601, 1570, 1509, 1249 and 1170cm- ¹ ; λ _mav (EtOH) 294 (ε _m 17,300) and 232nm (7,400); δ _H (CD3OD) 0.95

(3H, d, J 7.1Hz, 17-H ₃ ), 1.2 (3H, d, J 6.4Hz, I4-H3) , 1.4 (IH, m, 12-H) , 1.7 (IH, bt, J 6.8Hz, 9-H ₂ ) , 2.0 (IH, m, 8-H) , 2.2 (3H, s, I5-H3), 2.4 (IH, dd, J 9.6, 14.4Hz, 4-H), 2.75 (2H, m, 4-H and 11-H) , 2.85 (IH, dt, J 2.2, 5.8Hz, 510-H) , 3.4-4.0 (6H, m, 5,6,7,13-H and 16-H ) , 3.7 (3H, s, OMe) , 6.85 (IH, s, 2-H) and 7.0, 7.95 (4H, A ₂ B ₂ q, J 8.8Hz, Ar-H) ; m/z _. 434 (M ⁺ , 25%), 416 (20), 398 (25) and 135 (100) . (Found: M ⁺ , 434.2302. C ₂₄ H ₃₄ 0 ₇ requires 434.2305) .

0 Example 2

m-Methoxyphenyl-l-normon-2-yl ketone

a) N-Methoxy-N-methylmonamide

To monic acid A (6.88g, 20mmol) in dry tetrahydrofuran (50ml) at 0°C was added triethylamine (3.5ml, 25mmol) and isobutyl chloroformate (2.6ml, 20mmol) . The mixture was stirred at 0°C for 30min then a further portion of triethylamine (2.8ml, 20mmol) and also N,0- dimethylhydroxylamine hydrochloride (1.95g, 20mmol) were added. The mixture was allowed to warm to 20°C and stirred at this temperature for 2 h.

The solid was removed by filtration, and washed with tetrahydrofuran. The combined filtrates were evaporated under reduced pressure and the residue purified by chromatography on silica gel, eluting with methanol in dichloromethane (0-10%) , to give pure N-methoxy-Nmethylmonamide as a gum, (3.6g, 46%); δ _H (CD3OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.5Hz, I4-H3) , 1.4 (IH, m, 12-H), 1.7 (2H, m, 9-H ₂ ), 1.95 (IH, m, 8-H), 2.1 (3H, s, I5-H3), 2.2 (IH, dd, J 9.6, 14.5Hz, 4-H), 2.65 (IH, bd, J 14.5Hz, 4-H), 2.7 (IH, dd, J 2.2, 7.7Hz, 11-H), 2.8 (IH, dt, J 2.2, 5.7Hz, 10-H), 3.2 (3H, s, NMe), 3.3-3.9 (6H,

m, 5 , 6, 7, 13-H and 16-H ₂ ) , 3 . 9 (3H, s , OMe) and 6.2 ( IH, bs , 2-H) .

b) N-Methoxy-N-methyl 6,7,13-Tris-trimethylsilylmonamide

The above monamide (l.Q5g, 2.7mmol) was dissolved in tetrahydrofuran (20ml) and triethylamine (2.8ml, 20mmol) , chlorotrimethyl silane (1.9ml, 15mmol) and 4dimethylaminopyridine (20mg) added. The mixture was stirred at 20°C for Ih, then diluted with diethyl ether and filtered. The solid was washed with diethyl ether and evaporated under reduced pressure. The resultant gum was taken up in diethyl ether, refiltered and evaporated to give N-methoxy-N-methyl 6,7, 13-0-tristrimethylsilylmonamide as a colourless gum (1.55g, 95%).

c) m-Methoxyρhenyl-l-normon-2-yl ketone

To a solution of m-bromoanisole (0.187g, Immol) in tetrahydrofuran (4ml) at -78°C under nitrogen was added a solution of n-butyllithium (0.66ml, 1.5M in hexane) . The mixture was stirred at -78°C for 30min, when to the resulting white slurry was added dropwise a solution of the protected monamide from part b) (0.6g, lmmol) in tetrahydrofuran (3ml) . After stirring at -78°C for 15min saturated aqueous ammonium chloride (2ml) was added, and the mixture warmed to 20°C. Diethyl ether was added and the organic layer was washed with saturated aqueous sodium chloride, dried (MgSO^) and evaporated under reduced pressure to a pale yellow gum (0.73g) .

This material was taken up in tetrahydrofuran (20ml) and 0.4M hydrochloric acid (4ml) was added. After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was added, and the mixture extracted with ethyl acetate. The

organic layer was dried (MgS0 ₄ ), evaporated and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title compound as a colourless gum, (0.168g, 39%); ^v _maχ (KBr) 5 3434, 3075, 2969, 2927, 1656, 1595 and 1265cm- ¹ ; λ _maχ (EtOH) 312 (^ 2,900), 265 (13,000) and 218nm (16,500); δ _H (CD3OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.5Hz, I4-H3) , 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.3Hz, 9-H ₂ ) , 2.0 (IH, , 8-H), 2.2 (3H, s, 15-H ₃ ) , 2.35 (lH,dd, J 9.6, 14.4Hz, 4-H),

102.75 (3H, m, 10, 11, and 4-H), 3.4-4.0 (6H, m, 5, 6, 7, 13-H and 16-H ₂ ), 3.8 (3H, s, OMe), 6.85 (IH, s, 2-H) , 7.15 (IH, , Ar-4-H) and 7.4-7.6 (3H, m, Ar-2, 5 and 6-H) ; m/z. 434 (M ⁺ , 2%), 416 (5) and 135 (100) . (Found: 434.2302. ₂ H ₃₄ 0 ₇ requires 434.2305) .

15

Example 3

3, -Methylenedioxyρhenyl-l-normon-2-yl ketone

0 a) 6,7,13-0-Tris (tertbutyldimethylsilyl)monic acid

Monic acid A (11.41g, 33.2mmol), t.-butyldimethylsilylchloride (25.Og, 166mmol) and imidazole (22.55g, 332mmol) in N,N-dimethylformamide (50ml) were 5 heated to 70°C for 40h. Water (200ml) was added and the solution extracted with ethyl acetate (3 x 200ml) and the extracts dried (MgS0 ₄ ) . After evaporation to dryness under reduced pressure the crude products were dissolved in methanol (360ml) , THF (125ml) and water (125ml) . Potassium 0 carbonate (14.68g, 106mmol) was added and the solution stirred at room temperature for lh. The products were evaporated to low volume under reduced pressure, water

(200ml) added and 5N hydrochloric acid added until pH 3 was reached. Extraction with dichloromethane (3 x 750ml) , 5 drying (MgS0 ₄ ) and purification by flash chromatography

using 0-15% ethyl acetate in hexane gave 6-,7-,13-O-tris (tert butyldimethylsilyl)monic acid (14.16g, 62%) as a white solid; V _maχ (KBr) 3420, 3200-2200 (br) , 2929, 2857, 1693,

1638, 1254, 1122, 1085, 837cm- ¹ ; 5 _R (CDCI3) 0.04-0.10 (18H, 5 m, 6 x SiCH ₃ ), 0.89-0.93 (30H, m, 17-H ₃ and 3 x ^t Bu) , 1.19 (3H, d, J 6.3Hz, I4-H3), 1.29-1.43 (IH, m, 12-H), 1.73-1.97 (3H, m, 8-H and 9-H ₂ ) , 1.98-2.10 (IH, m, 4-H), 2.53-2.64 (IH, m, 4-H), 2.64-2.77 (2H, m, 10 and 11-H), 3.38 (IH, dd,

J 9.1 and 1.8Hz, 6-H) , 3.55 (IH, d, J 11.3Hz, 16-H) , 10 3.82-3.95 (4H, m, 5,7,13 and 16-H), 5.76 (IH, s, 2-H) ; δ _c (CDCI3) , -4.9 (SiCH ₃ ), -4.6 (SiCH ₃ ) , -4.5 (SiCH ₃ ) , -4.3 (SiCH ₃ ), -4.2 (SiCH ₃ ) , -3.3 (SiCH ₃ ) , 12.7 (C-17) , 18.0 [C(CH ₃ ) ₃ ], 18.1 [C(CH ₃ ) ₃ ], 18.1 [C(CH ₃ ) ₃ ], 19.1 (C-15), 20.9 (C-14), 25.7 ([C(CH ₃ ) ₃ ], 25.9 [C(CH ₃ ) ₃ ], 26.1 [C(CH ₃ ) ₃ ], 1531.9 (C-9), 42.1 (C-8) , 43.0 (C-12) , 43.5 (C-4) , 55.5

(C-10), 59.1 (C-ll) , 65.4 (C-16) , 70.3 (C-6) , 70.8 (C-7) ,

73.4 (C-13), 74.1 (C-5) , 116.6 (C-2) , 160.5 (C-3) , 171.7 (C-l); m/z. (FAB 3-NOBA/Na) , 731 [M-H + 2Na) ⁺ , 100%], 709 t(MNa) ⁺ , 27%]. 20 b) N-Methoxy-N-methyl

6,7, 13-O-tris-t-butyldimethylsilylmonamide

A solution of tris-t-butyldimethylsiyl protected monic acid 25 (1.37g, 2mmol) in dry dichloromethane (10ml) at 5°C was sequentially treated with triethylamine (0.3ml, 2.2mmol) and iso-butylchloroformate (0.27ml, 2.1mmol). After 30 min triethylamine (0.3ml, 2.2mmol) and then

N,p_-dimethylhydroxylamine hydrochloride (200mg, 2.1mmol) was 30 added. After 2 h, the mixture was diluted with dichloromethane, washed with brine, then dried and evaporated. Chromatography on silica eluting with ethyl acetate/hexane mixtures gave the title compound as a crystalline solid (l.lδg, 79%); δ (CDCI3) (inter alia) 1.20 35 (3H, d, J 6.4Hz, I4-H3), 2.14 (3H, s, I5-H3) , 3.20 (3H, S, NMe), 3.67 (3H, s, NOMe) , 6.18 (IH, s, 2-H) .

-35- c) 6,7, 13-0-Tris-t-butyldimethylsilyl- (3,4- methylenedioxyl)phenyl-l-normon-2-yl ketone

To 4-bromo-l,2-methylenedioxybenzene (0.201g, lmmol) in tetrahydrofuran (4ml) at -78°C under nitrogen was added dropwise n-butyllithium (0.66ml, 1.5M in hexane) . The mixture was stirred at -78°C for 15min, after which time th tris-protected monamide above (0.73g, lmmol) in tetrahydrofuran (3ml) was added dropwise. After stirring for 15min. at -78°C saturated aqueous ammonium chloride (5ml) was added and the mixture warmed to 20°C. Diethyl ether was added and the organic layer was washed with saturated brine, dried (MgSO^) and concentrated by evaporation under reduced pressure. The residue was purified by chromatography on silica gel eluting with ethyl acetate in hexane (5-20%) to give

6, 7, 13-O-tris (tertbutyldimethylsilyl) - (3, 4-methyl- enedioxy)phenyl-l-normon-2-yl ketone as a colourless gum (0.232g, 29%); δ _R (CD3OD) (inter alia) 0.05-0.15 (18H, m, SiMe), 0.9 (30H, m, S^Bu and 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 2.1 (3H, s, I5-H3) , 2.2 (IH, dd, J 10.8, 13.7Hz, 4-H), 2.7 (IH, d, J 13.7Hz, 4-H), 2.8 (2H, m, 10- and 11-H), 6.05 (2H, s, OCH ₂ 0) , 6.75 (IH, s, 2-H), 6.7 (IH, d, J 8.2Hz, Ar-5-H) , 7.4 (IH, d, J 1.5Hz, Ar-2-H) and 7.6 (IH, dd, J 1.5, 8.2Hz, Ar-6-H) .

d) 3, -Methylenedioxyphenyl-l-normon-2-yl ketone

This protected ketone was dissolved in tetrahydrofuran (5ml) and treated at 20°C for 24h with tetrabutylammonium fluoride trihydrate (0.8g, 9eq) . The reaction mixture was diluted with diethyl ether and washed with water and brine. The combined aqueous washings were re-extracted with ethyl acetate, and the combined organic layers were dried (MgS0 ₄ ) evaporated and purified by chromatography on silica gel eluting with methanol in dichloromethane (0-5%) to give the title compound as a gum (60mg, 53%); ^v _maχ (KBr) 3430, 2923,

1645, 1603, 1445 and 1254cm- ¹ ; λ _maχ (EtOH) 319 (ε _m 9,900), 264 (9,300) and 238nm (14,000); δ _R 0.95 (3H, d, J 7.1Hz, 17-H ₃ ), 1.2 (3H, d, J 6.5Hz, I4-H3), 1.4 (IH, m, 12-H), 1.7 (2H, t, J 7.7Hz, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.15 (3H, s, 5 I5-H3), 2.35 (IH, dd, J 9.5, 14.4Hz, 4-H), 2.7 (3H, m, 4 , 10 and 11-H), 3.4-4.0 (6H, m, 5,6,7,13-H and 16-H ₂ ) , 6.05 (2H, S, OCH ₂ 0) , 6.8 (IH, s, 2-H), 6.9 (IH, d, J 8.1Hz, Ar-5-H) , 7.4 (IH, d, J 1.7Hz, Ar-2-H) and 7.6 (IH, dd, J 1.7, 8.1Hz, Ar-6-H); m/z.448 (M ⁺ , 2%), 149 (100). (Found: 448.2106. 0 C24H ₃₂ 0 ₈ requires 448.2097) .

Example 4 p-Methylthiophenyl-l-normon-2-yl-ketone

15 To a solution of p_-bromothioanisole (0.305g, 1.5mmol) in tetrahydrofuran (5ml) at -78°C under nitrogen was added a solution of n-butyllithium (1.0ml, 1.5M in hexane) . The mixture was stirred at -78°C for 15min, when to the resulting white slurry was added dropwise a solution of the 0 protected monamide prepared as in Example 2b (0.6g, lmmol) in tetrahydrofuran (4ml) . After stirring at -78°C for 15min acetic acid (0.12g) was added, and the mixture warmed to 20°C. Diethyl ether was added and the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ )

25 and evaporated under reduced pressure to a pale yellow gum (0.80g) .

This material was taken up in tetrahydrofuran (20ml) and 0.4M hydrochloric acid (4ml) was added. After 2.5min at 3020°C saturated aqueous sodium hydrogen carbonate (5ml) was added, and the mixture extracted with ethyl acetate. The organic layer was dried (MgSO^) , evaporated and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title

compound as a colourless gum, (0.26g, 57%); v _maχ (KBr) 3427, 2969, 2922, 1649, 1602, 1588, 1551, 1251 and 1095cm- ¹ ; λ _maχ (EtOH) 332 (ε _m 18,500) and 247nm (8,800); δ _H (CD ₃ OD) 0.95 (3H, d, J 7.0Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.5Hz, 14-H ₃ ) , 1.4 5 (IH, m, 12-H), 1.7 (2H, t, J 6.5Hz, 8-H ₂ ) ; 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ), 2.35 (IH, dd, J 9.5, 14.3Hz, 4-H), 2.5 (3H, s, SMe), 2.75 (3H, m, 10, 11, and 4-H), 3.4-4.0 (6H, , 5, 6, 7, 13-H and 16-H ₂ ) , 6.85 (IH, s, 2-H), 7.3 and 7.9 (4H, A ₂ B ₂ q, J 8.6Hz, Ar-H) ; m/z.450 (M ⁺ , 3%) and 151 (100) . 10 (Found: 450.2085. C ₂₄ H ₃₄ 0gS requires 450.2076) .

Example 5

p-Methylsulphinyl-l-normon-2-yl ketone

15

To the p_-methylthiophenyl ketone from Example 4, (0.164g, 0.36mmol) in dichloromethane (10ml) was added saturated aqueous sodium hydrogen carbonate (5ml) . The mixture was cooled to 0°C and m-chloroperbenzoic acid (0.071g) added.

20 After stirring at 0°C for 15min the mixture was diluted with dichloromethane (50ml) and the organic layer dried (MgS0 ₄ ) and evaporated. The residue was purified by chromatography on silica eluting with methanol in dichloromethane (0-8%) . The title product was obtained as a white foam, (O.llβg,

25 68%); V _maχ (KBr) 3421, 2969, 2920, 1657, 1608, 1241 and

1036cm- ¹ ; λ _maχ (EtOH) 278 (ε _m 17,730) and 204nm (19,250); δ _R (CD3OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.5Hz, I4-H3), 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.6Hz, 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ) , 2.4 (IH, dd, J 7.6,

30 14.3Hz, 4-H), 2.75 (3H, m, 4, 10 and 11-H), 2.8 (3H, s,

SMe), 3.4-4.0 (6H, m, 5, 6, 7, 13-H and 16-H ₂ ) , 6.9 (IH, s, 2-H), 7.8 and 8.1 (4H, A ₂ B ₂ q, J 8.4Hz, Ar-H); m/z. (FAB, 3-NOBA/Na) 489 (MNa ⁺ ) .

Example 6

p-Hydroxyphenyl-l-normon-2-yl ketone

5 To a solution of p-bromophenoxytrimethylsilane (0.22g, 0.9mmol) in tetrahydrofuran (3ml) at -78°C under nitrogen was added a solution of n-butyllithium (0.6ml, 1.5M in hexane) . The mixture was stirred at -78°C for 15min, when to the resulting solution was added dropwise a solution of 0 the protected monamide prepared as in Example 2b (0.4g,

0.66mmol) in tetrahydrofuran (3ml). After stirring at -78°C for 75min acetic acid (0.09g) was added, and the mixture warmed to 20°C. Diethyl ether was added and the organic layer was washed with saturated aqueous sodium chloride, 5 dried (MgS0 ) and evaporated under reduced pressure to a pale yellow gum (0.50g). Chromatography on silica eluting with diethyl ether in dichloromethane (0-12%) gave partially purified tris-protected p-hydroxyphenyl-lnormon-2-yl ketone (0.08g) . This material was taken up in tetrahydrofuran 0 (20ml) and 0.4M hydrochloric acid (4ml) was added. After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was added, and the mixture extracted with ethyl acetate. The organic layer was dried (MgS0 ₄ ) , evaporated and the residue purified by chromatography on silica gel

25 eluting with methanol in dichloromethane (0-10%) to give the title compound as a colourless gum, (0.025g, 9%); v _maχ (KBr) 3421, 2969, 2920, 1657, 1608, 1403, 1241 and 1036cm- ¹ ; λ _m ie _a lA _v

(EtOH) 274 (ε _m 17,730) and 205nm (19,250); δ _R (CD ₃ OD) 0.9 (3H, d, J 7.0Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.5Hz, 14-H ₃ ) , 1.4

30 (IH, m, 12-H), 1.7 (2H, t, J 6.6Hz, 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.15 (3H, s, 15-H ₃ ) , 2.35 (IH, dd, J 9.5, 14.3Hz, 4-H), 2.7 (3H, m, 4, 10 and 11-H), 3.4-4.0 (6H, m, 5, 6, 7, 13-H and 16-H ₂ ), 6.8 (IH, s, 2-H), 6.8 and 7.85 (4H, A ₂ B ₂ q, J 8.8Hz, Ar-H); m/z.420 _(M ⁺ , 5%), 402 (20) and 121 (100) . (Found:

35420.2143. C ₂₃ H ₃₂ 0 ₇ requires 420.2148).

Example 7

4-Dimethylaminophenyl-l-normon-2-yl ketone

5 To a solution of 4.-bromo-N,N-dimethyl-aniline (0.22g, 1.lmmol) in tetrahydrofuran (4ml) at -78°C under nitrogen was added a solution of n-butyllithium (0.66ml, 1.5M in hexane) . The mixture was stirred at -78°C for 15min, when to the resulting white slurry was added dropwise a solution

10 of the protected monamide prepared as in Example 2b (0.4g, 0.66mmol) in tetrahydrofuran (3ml) . After stirring at -78°C for 15min acetic acid (0.07g) was added and the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure to a

15 pale yellow/green oil (0.46g) .

This material was taken up in tetrahydrofuran (20ml) and 0.4M hydrochloric acid (4ml) was added. After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was

20 added, and the mixture extracted with ethyl acetate. The organic layer was dried (MgS0 ₄ ), evaporated and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title compound as a yellow/green foam (0.135g, 45%); ^v _maχ (KBr)

253422, 2968, 2910, 1645, 1602, 1595, 1370 and 1266cm- ¹ ; λ _maχ (EtOH) 259 (ε _m 23,150), 250 (13,000) and 205nm (17,870); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.0Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 14-H ₃ ), 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.4Hz, 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.1 (3H, s, 15-H ₃ ) , 2.35 (IH, dd, J 9.4,

30 14.4Hz, 4-H), 2.75 (3H, m, 10, 11, and 4-H), 3.05 (6H, s, NMe ₂ ), 3.4-4.0 (6H, , 5, 6, 7, 13-H and 16-H ₂ ) , 6.8 (IH, s, 2-H), 6.7 and 7.9 (4H, A ₂ B ₂ q, J 9.1Hz, Ar-H); m/z. 447 (M ⁺ , 5%), 163 (30) and 148 (100) . (Found: 447.2626. ₂₅ H ₃₇ NOg requires 447.2621).

35

Example 8

4-Methylsulphonylphenyl-l-normon-2-yl ketone

To the £-methylthiophenyl ketone from Example 4 (0.037g, 0.082mmol) in dichloromethane (2ml) was added saturated aqueous sodium hydrogen carbonate (2ml) . The mixture was cooled to 0°C and m-chloroperbenzoic acid (0.035g) added. After stirring at 0°C for 90min the mixture was diluted with dichloromethane (20ml) and the organic layer dried and evaporated. The residue was purified by chromatography on silica eluting with methanol in dichloromethane (0-6%) . The title product was obtained as a white foam, (0.022g, 55%); V _maχ (KBr) 3453, 2970, 2924, 1661, 1609, 1314, 1297, 1153 and 776cm- ¹ ; ^ _ax (EtOH) 269 (ε _m 14,070) and 251nm (15,420); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.5Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.6Hz, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.25 (3H, s, 15-H ₃ ) , 2.4 (IH, dd, J 9.6, 14.4Hz, 4-H), 2.7 (3H, , 4, 10 and 11-H), 3.15 (3H, s, S-Me) , 3.4-4.0 (6H, m, 5, 6, 7, 13-H and 16-H ₂ ) , 6.9 (IH, s, 2-H), 8.05 and 8.15 (4H, A ₂ B ₂ q, J 8.4Hz, Ar-H); m/z.482 (M ⁺ , 0.2%) , 464 (2) , 183 (100) .

Example 9

4-Cyanophenyl-l-normon-2-yl ketone

To a solution of p_-bromo-benzonitrile (0.245g, 1.5mmol) in tetrahydrofuran (4ml) at -78°C under nitrogen was added a solution of n-butyllithium (1.0ml, 1.5M in hexane). The mixture was stirred at -78°C for 2min, when to the resulting red solution was added dropwise a solution of the protected monamide prepared as in Example 2b (0.6g, l.Ommol) in tetrahydrofuran (5ml) . After stirring at -78°C for 15min

acetic acid (0.07g) ,diethyl ether and water were added, the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure to a yellow oil. 5

This material was taken up in tetrahydrofuran (20ml) and 0.4M hydrochloric acid (4ml) was added. After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was added, and the mixture extracted with diethyl ether. The

10 organic layer was dried (MgS0 ₄ ) , evaporated and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title compound as a colourless foam (0.260, 60%); v _maχ (KBr) 3436, 2969, 2924, 2230, 1660, 1611, and 1243cm- ¹ ; λ _maχ (EtOH)

15259nm (ε _m 19,300); δ _H (CD ₃ 0D) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.5Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.5Hz, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.25 (3H, s, 15-H ₃ ) , 2.4 (IH, dd, J 9.5, 14.3Hz, 4-H), 2.7-2.9 (3H, m, 4-, 10- and 11-H), 3.4-3.9 (6H, m, 5, 6,7, 13-H and 16-H ₂ ) , 6.9 (IH, s, 02-H), 7.85 and 8.1 (4H, A ₂ B ₂ q, J 8.5Hz, Ar-H ₄ ); m/z _. 429 (M ⁺ , 1%), 411 (4) and 130 (100). (Found: 429.2152. C ₂₄ H ₂₁ 0g requires 429.2151) .

Example 10 5

3-Fluoro-4-methoxyρhenyl-l-normon-2-yl ketone To a solution of 4-bromo-2-fluoro anisole (0.41g, 2.0mmol) in tetrahydrofuran (8ml) at -78°C under nitrogen was added a solution of n-butyllithium (1.15ml, 1.7M in hexane) . The 0 mixture was stirred at -78°C for 15min, when to the resulting white slurry was added dropwise a solution of the protected monamide prepared as in Example 2b (0.6g, l.Ommol) in tetrahydrofuran (5ml) . After stirring at -78°C for 15min acetic acid (0.l2g) diethyl ether and water were added, the 5 organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure.

-42-

This material was taken up in tetrahydrofuran (20ml) and 0.4M hydrochloric acid (4ml) was added- After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (10ml) was added, and the mixture extracted with diethyl ether. The 5 organic layer was dried (MgS0 ₄ ) , evaporated and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title compound as a colourless foam (0.3g, 66%); v _maχ (KBr) 3359, 2970, 2913, 1658, 1612, 1515, 1438 and 1266cm- ¹ ; λ _maχ (EtOH)

10295nm (ε _m 16,000); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.0Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.5Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.7Hz, 9-H ₂ ), 1.95 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ) , 2.35 (IH, dd, J 14.4, 9.6Hz, 4-H), 2.7-2.9 (3H, m, 10, 11 and 4-H), 3.4-3.9 (6H, m, 5, 6,7,13-H and 16-H ₂ ) , 3.95, (3H, s,

15 OMe) , 6.85 (IH, s, 2-H), 7.15 (IH, t, J 8.4Hz, Ar-5-H) , 7.7 (IH, dd J 8.4, 2.1Hz, Ar-6-H) and 7.8 (IH, d, J 8.7Hz, Ar-2-H) ; m/z.452 (M ⁺ , 2%), 153 (100). (Found: 452.2213. ^C 24 ^H 33°7 ^F requires 452.2210) .

20 Example 11

2-Methoxyphenyl-l-normon-2-yl ketone

To a solution of 2-bromo anisole (0.247ml, 2.0mmol) in 25 tetrahydrofuran (5ml) at -78°C under nitrogen was added a solution of n-butyllithium (1.0ml, 1.5M in hexane). The mixture was stirred at -78°C for 15min, when to the resulting solution was added dropwise a solution of the protected monamide prepared as in Example 2b (0.6g, l.Ommol) 30 in tetrahydrofuran (5ml) . After stirring at -78°C for 15min acetic acid (0.07g) diethyl ether and water were added, the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure. 35

This material was taken up in tetrahydrofuran (20ml) and 0.4M hydrochloric acid (4ml) was added.. After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was added, and the mixture extracted with diethyl ether. The organic layer was dried (MgS0 ), evaporated and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title compound as a colourless foam (0.293g, 67%); v _maχ (KBr) 3429, 2968, 2930, 1655, 1604, 1458, 1287 and 1246cm- ¹ ; λ _maχ (EtOH) 310 (ε _m 4,400), 264 (14,800) and 214nm (13,200); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 14-H ₃ ), 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.4Hz, 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ) , 2.3 (IH, dd, J 9.6, 14.5Hz, 4-H), 2.75 (3H, m, 10, 11 and 4-H), 3.4-4.0 (6H, m, 5, 6,7, 13-H and 16-H ₂ ) , 3.9 (3H, s, OMe), 6.7 (IH, s, 2-H), 7.0-7.1 (2H, m, Ar-5-and-3-H) and 7.5 (2H, m, Ar-4-and-6-H) ; m/z.434 (M ⁺ , 2%) and 135 (100). (Found: 434.2302. C ₂₄ H ₃₄ 0 ₇ requires 434.2305).

Example 12

4-Hvdroxymethylphenyl-l-normon-2-yl ketone

To 4-bromobenzyl alcohol (1.87g, lOmmol) in tetrahydrofuran (20ml) was added triethylamine (2.2ml, 15mmol) and chlorotrimethyl silane (1.9ml, 15mmol) . After lOmin the mixture was diluted with diethyl ether, filtered, and evaporated. The residue was taken up in hexane, refiltered and evaporated to a yellow solid, 2.57g.

To a solution of this silyl ether in tetrahydrofuran (40ml) at -78°C under nitrogen was added a solution of n-butyllithium (6.67ml, 1.5M in hexane) . The mixture was stirred at -78°C for 15min, when to the resulting white slurry was added dropwise a solution of the protected

monamide prepared as in Example 2b (4.0g, 6.67mmol) in tetrahydrofuran (30ml) . After stirring at -78°C for 15min acetic acid (0.7g) diethyl ether and water were added, the organic layer was washed with saturated aqueous sodium 5 chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure.

This material was taken up in tetrahydrofuran (50ml) and 0.4M hydrochloric acid (10ml) was added. After 2.5min at

1020°C saturated aqueous sodium hydrogen carbonate (10ml) was added, and the mixture extracted with diethyl ether. The organic layer was dried (MgS0 ₄ ) , evaporated and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title

15 compound as a white foam (0.61g, 21%); v _maχ (KBr) 3412, 2968, 1668, 1609, 1249, and 1051cm- ¹ ; λ _maχ (EtOH) 269.5nm (ε.^ 20,200); δ _H (CD3OD) 0.95 (3H, d, J 7Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.5Hz, I4-H3), 1.4 (IH, m, 12-H), 1.7 (2H, m, 9-H ₂ ) , 2.2 (3H, s, I5-H3), 2.4 (IH, dd, J 9.6, 14.4Hz, 4-H),

20 2.7-2.8 (3H, m, 4, 10 and 11-H), 3.4-3.9 (6H, m, 5, 6,7, 13-H and 16-H ₂ ) , 4.65 (2H, s, hydroxymethyl) , 6.9 (IH, s, 2-H), 7.45 and 7.9 (4H, ₂ B ₂ q, J 8.1Hz, Ar-H ₄ ) ; δ _c (CD3OD) 12.3 (C-17), 20.4 (C-14 and 15), 33.0 (C-9) , 41.7 (C-8) , 43.7 (C-12), 44.3 (C-4) , 56.9 (C-10) , 61.3 (C-ll) , 64.6 (CH ₂ OH) ,

25 66.4 (C-16) , 70.0, 70.7 and 71.6 (C-6, 7 and 13), 76.4

(C-5), 123.7 (C-2), 127.7 (Ar-C-3 and 5), 129.5 (Ar-C-2 and 6), 139.3 (Ar-C-1), 148.0 (Ar-C-4) , 158.8 (C-3) and 193.4 (C-l) ; m/z.434 (M ⁺ , 3%) and 135 (100) . (Found: M ⁺ , 434.2302. ^C 24 ^H 34°7 req ires 434.2305) .

30

-45- Example 13

4-Formylphenyl-l-normon-2-yl ketone

To the hydroxymethylphenyl ketone prepared as in Example 12 (0.276g, 0.64mmol) in tetrahydrofuran (30ml) at 5°C was added manganese dioxide (0.9g) and the mixture stirred at 5°C for 65h. The oxidant was removed by filtration and washed with tetrahydrofuran and methanol. The combined solutions were evaporated and the residue purified by chromatography on silica gel eluting with 0-5% methanol in dichloromethane to give the title aldehyde (0.15g, 54%); V _maχ (KBr) 3435, 2969, 2922, 1703, 1656, 1600, 1241, 1207 and 818cm- ¹ ; λ _maχ (EtOH) 269.5nm (ε _m 23,800); δ _R (CD3OD) 0.95 (3H, d, J 7.0Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, m, 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.2 > x 3H, s,hemiacetal-15-H ₃ ) , 2.25 (*έ x 3H, s, aldehyde-15-H ₃ ) , 2.4 (IH, m, 4-H), 2.7-2.8 (3H, m, 4, 10 and 11-H), 3.4-3.9 (6H, m, 5, 6,7, 13-H and 16-H ₂ ), 5.6 Vz x IH, s, hemiacetal-H) , 6.9 (*έ x IH, s, hemiacetal-2-H) , 6.95 (^ x IH, s, aldehyde-2-H) , 7.6, 7.8 (^ X 4H, A ₂ B ₂ q, J 8.2Hz, hemiacetal-Ar-H ₄ ) , 0.8 and 8.1 (^ x 4H, A ₂ B ₂ q, J 8.3Hz, aldehyde-Ar-H ₄ ) , 10.1 x IH, s, aldehyde-H) ; m/z. 432 (M ⁺ , 1%), 414 (2), 200 (60) and 133 (100) . (Found: M ⁺ , 432.2158. C ₂₄ H ₃₂ 0 ₇ requires 432.2148).

Example 14

4-Acetylphenyl-l-normon-2-yl ketone

To 4-bromoacetophenone (1.2g, 6mmol) in diethyl ether (15ml) at 0°C was added triethylamine (1.26ml, 9mmol) and trimethylsilyl triflate (1.35ml, 7mmol) . The mixture was allowed to warm to 20°C over 30min. The mixture was diluted

with hexane and the solution decanted from an oil which was triturated with further hexane. The combined solutions were washed with ice cold water and brine. After drying and evaporation the required enol ether was obtained as an oil 5 (1.35g); δ _R (CC1 ₄ ) , 0.4 (9H, s, SiMe ₃ ) , 4.55 and 5.0 (2H, ABq, J 2Hz, =CH ₂ ), 7.6 (4H, S, ArH) .

To a solution of this enol ether (0.81g, 3.0mmol) in tetrahydrofuran (12ml) at -78°C under nitrogen was added a

10 solution of n.-butyllithium (2.0ml, 1.5M in hexane). The mixture was stirred at -78°C for 15min, when to the resulting solution was added dropwise a solution of the protected monamide prepared as in Example 2b (1.2, 2.0mmol) in tetrahydrofuran (10ml) . After stirring at -78°C for

15 15min acetic acid (0.20g) diethyl ether and water were added, the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure.

20 This material was taken up in tetrahydrofuran (40ml) and 0.4M hydrochloric acid (8ml) was added. After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (8ml) was added, and the mixture extracted with ethyl acetate. The organic layer was dried (MgS0 ₄ ) , evaporated and the residue

25 purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title compound as a white foam (0.325g, 36%); v _maχ (KBr) 3437, 2965, 2923, 1685, 1658, 1600, 1265 and 1241cm- ¹ ; λ _m=v (EtOH) 269nm (ε _m 24,800); δ _R (CDCI3) 0.95 (3H, d, J 7.0Hz, 17-H ₃ ) ,

30 1.2 (3H, d, J 6.3Hz, 14-H ₃ ) , 1.75 (2H, m, 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.25 (3H, s, 15-H ₃ ) , 2.4 (IH, dd, J 9.4, 14.3Hz, 4-H), 2.65 (3H, s, acetyl-2-H ₃ ) , 2.7-2.8 (3H, m, 4, 10 and 11-H), 3.5-4.0 (6H, m, 5, 6, 7, 13-H and 16-H ₂ ) , 6.8 (IH, s, 2-H), 8.0 (4H, s, Ar-H ₄ ); m/z.446 (M ⁺ , 1%), 428 (1) and (147 (100) .

35 (Found: M ⁺ , 446.2296. C ₂₅ H ₃₄ 0 ₇ requires 446.2305) .

Example 15

4- (2-Hvdroxyethoxyiminomethyl)ρhenyl-l-normon-2-yl ketone;

To the 2-trimethylsilyloxyethoxime of 4-bromo benzaldehyde (0.474g, 1.5mmol) (prepared by alkylating the tetrabutylammonium salt of the oxime of p-bromobenzaldehyde with iodoethanol in tetrahydrofuran followed by treatment with chlorotrimethyl silane and triethylamine) in tetrahydrofuran (5ml) at -90°C under nitrogen was added a solution of n-butyllithium (1.0ml, 1.5M in hexane) . The mixture was stirred at -90°C for 5min, when to the resulting orange-brown solution was added dropwise a solution of the protected monamide prepared as in Example 2b (0.6g, l.Ommol) in tetrahydrofuran (5ml) . After stirring at -90 to -78°C for 15min acetic acid (0.10g) was added, the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure.

This material was taken up in tetrahydrofuran (20ml) and 0.4M hydrochloric acid (4ml) was added. After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was added, and the mixture extracted with diethyl ether. The organic layer was dried (MgS0 ₄ ), evaporated and the residue purified by chromatography on silica gel eluting with ethyl acetate to give the title compound as colourless crystals from ethyl acetate/hexane (0.112g, 23%); mp 101-102°C; v _maχ (KBr) 3528, 3435, 2967, 2913, 2888, 1647, 1607, 1591, 1251, 1114, 1051 and 1040cm- ¹ ! λ _maχ (EtOH) 298nm (ε _m 28,300); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 14-H ₃ ), 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.5Hz, 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ) , 2.4 (IH, dd, J 9.5 and 14.3Hz, 4-H), 2.7-2.8 (3H, m, 4-, 10- and 11-H), 3.4-3.9 (8H, m, 5-, 6-, 7- and 13-H, 16-H ₂ and ethoxy-2-H ₂ ) , 4.25 (2H, t, J 4.9Hz, ethoxy-l-H ₂ ) , 6.9 (IH, s, 2-H), 7.7 and

7.95 (4H, A ₂ B ₂ q, J 8.4Hz, Ar-H ₄ ) and 8.2 (IH, s, CH=N) ; m/z. (FAB, thioglycerol) 514 (MNa ⁺ ) and 492 (MH ⁺ ) .

Example 16 5

4- (4-Hvdroxybutoxyiminomethyl)phenyl-l-normon-2-yl ketone

To the 4-trimethylsilyloxybutoxime of 4-bromo benzaldehyde (0.474g, 1.5mmol) (prepared by alkylating the

10 tetrabutylammonium salt of the oxime of p_-bromobenzaldehyde with 4-iodo-l-trimethylsilyloxy butane in tetrahydrofuran and chromatography on silica gel, followed by treatment with chlorotrimethyl silane and triethylamine) in tetrahydrofuran (5ml) at -90°C under nitrogen was added a solution of

15 n-butyllithium (1.0ml, 1.5M in hexane) . The mixture was stirred at -90 to -78°C for 5min, when to the resulting orange solution was added dropwise a solution of the protected monamide prepared as in Example 2b (0.6g, l.Ommol) in tetrahydrofuran (5ml) . After stirring at -90°C for 15min

20 acetic acid (O.lOg) was added, the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure.

This material was taken up in tetrahydrofuran (20ml) and 250.4M hydrochloric acid (4ml) was added. After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was added, and the mixture extracted with diethyl ether. The organic layer was dried (MgS0 ₄ ) , evaporated and the residue purified by chromatography on silica gel eluting with ethyl 30 acetate to give the title compound as colourless foam, (0.175g, 34%); V _maχ (KBr) 3463, 2969, 2923, 2876, 1653, 1605, 1591, 1248, 1057, 1032 and 943cm_l. ^^ (EtOH) 299nm (ε _m 28,000); δ _H (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 14-H ₃ ), 1.4 (IH, m, 12-H), 1.6-1.8 (6H, m, 35 9-H ₂ and butoxy-2-and-3-H ₂ ) , 2.2 (3H, s, 15-H ₃ ) , 2.35 (IH,

dd, J 9.5 and 14.4Hz, 4-H), 2.7-2.8 (3H, m, 4-, 10- and 11-H), 3.4-3.9 (8H, m, 5-, 6-, 7- and 13-H, 16-H ₂ and butoxy-4-H ₂ ) , 4.2 (2H, t, J 6.3Hz, butoxy-l-H ₂ ) , 6.9 (IH, s, 2-H), 7.7 and 7.95 (4H, ₂ B ₂ q, J 8.4Hz, Ar-H ₄ ) and 8.15 (IH, s, CH=N) ; m/z _. (FAB, thioglycerol) 542 ⁽ MNa ⁺ ) and 520 (MH ⁺ ) .

Example 17

4- (1-Hydroxyethyl)phenyl-l-normon-2-yl ketone

To 4- (l-hydroxyethyl)bromobenzene (0.6g, 3mmol) in tetrahydrofuran (10ml) was added triethylamine (1ml, 7mmol) and chlorotrimethyl silane (0.76ml, 6mmol) . After 4h at 20°C the mixture ^' was diluted with diethyl ether and filtered. The solvent was removed and the residue taken up in hexane, refiltered and concentrated to an oil (0.8g) . To a solution of the above aryl bromide (0.63g, 2.3mmol) in tetrahydrofuran (10ml) at -78°C under nitrogen was added a solution of n-butyllithium (1.5ml, 1.5M in hexane) . The mixture was stirred at -78°C for 15min, when to the resulting solution was added anhydrous cerium (III) chloride (0.566g, 2.3mmol). After a further hour at -78°C a solution of the protected monamide prepared as in Example 2b (0.9g, 1.5mmol) in tetrahydrofuran (6ml)was added dropwise. After stirring at -78°C for 15min acetic acid (0.20g), diethyl ether and water were added, the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure.

This material was taken up in tetrahydrofuran (20ml) and 0. M hydrochloric acid (4ml) was added. After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was added, and the mixture extracted with diethyl ether. The organic layer was dried (MgS0 ₄ ), evaporated and the residue

-50- purified by chromatography on silica gel eluting with ethyl acetate to give the title compound as a white foam (0.143g, 21%); V_ _av (KBr) 3425, 2970, 1655, 1610, 1249 and 1091cm- ¹ ; λ _maχ (EtOH) 299nm (ε _m 21,200); δ _R (CD ₃ OD) 0.95 (3H, d, J 57.0Hz, 17-H ₃ ), 1.2 (3H, d, J 6.5Hz, 14-H ₃ ) , 1.45 (4H, m, 12-H and ethyl-2-H ₃ ) , 1.7 (2H, m, 9-H ₂ ) , 2.0 (IH, m, H-8) , 2.2 (3H, s, 15-H ₃ ), 2.35 (IH, dd, J 9.6, 14.4Hz, 4-H), 2.7-2.8 (3H, m, 4-, 10- and 11-H), 3.4-3.9 (6H, m, 5-, 6-, 7-, 13-H and 16-H ₂ )4.85 (IH, , ethyl-1-H) , 6.9 (IH, s, 102-H), 7.5 and 7.9 (4H, A ₂ B ₂ q, J 8.3Hz, Ar-H ₄ ) ; m/z. (FAB, 3-NOBA/Na) 471 (MNa ⁺ ) .

Example 18

15 l-Normon-2-yl-4- (ρyrrolidin-1-ylsulρhonyl)phenyl ketone

To 1- (4-bromoρhenyl) sulphonyl pyrrolidine (0.414g, 1.5mmol) (prepared from 4-bromophenylsulphonyl chloride and pyrrolidine in the presence of triethylamine followed by

20 crystallisation from dichloromethane/ hexane, mp 97-98°C) in tetrahydrofuran (8ml) at -78°C under nitrogen was added a solution of n-butyllithium (0.66ml, 1.5M in hexane) . The mixture was stirred at -78°C for 15min, when to the resulting orange solution was added

25 dropwise a solution of the protected monamide prepared as in Example 2b (0.6g, l.Ommol) in tetrahydrofuran (5ml)was added dropwise. After stirring at -78°C for 15min acetic acid (0.20g), diethyl ether and water were added, the organic layer was washed with saturated aqueous sodium chloride,

30 dried (MgS0 ₄ ) and evaporated under reduced pressure.

This material was taken up in tetrahydrofuran (20ml) and 0.4M hydrochloric acid (4ml) was added. After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was 35 added, and the mixture extracted with ethyl acetate. The

-51- organic layer was dried (MgS0 ₄ ) , evaporated and the residue purified by chromatography on silica ge ^' l eluting with methanol in dichloromethane (0-5%) to give the title compound as a colourless foam (0.255g, 49%); v _maχ (KBr) 53411, 2971, 2929, 2877, 1660, 1609, 1346, 1240 and 1161cm- ¹ ; λ _maχ (EtOH) 273nm (ε _m 20,100); δ _R (CD ₃ 0D) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ), 1.2 (3H, d, J 6.4Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (6H, m, pyrrolidine 2- and 3-H ₂ and 9-H ₂ ), 2.0 (IH, m, 8-H), 2.25 (3H, s, 15-H ₃ ) , 2.4 (IH, dd, J 9.5 and 10 14.3Hz, 4-H), 2.7-2.8 (3H, m, 4-, 10- and 11-H), 3.25 (4H, m, pyrrolidine 1- and 4-H ₂ ) , 3.6-3.9 (6H, m, 5, 6, 7, 13-H and 16-H ₂ ), 6.95 (IH, s, 2-H), 7.9 and 8.1 (4H, A ₂ B ₂ q, J 8.4Hz, Ar-H ₄ ); m/z. (FAB, 3-NOBA/Na) 560 (MNa ⁺ ) .

5 Example 19

4-(2-Hydroxyethoxy)phenyl-l-normon-2-yl ketone

4-Bromophenol (3.4g, 20mmol) was dissolved in a methanol 0 solution of tetrabutyl ammonium hydroxide (1M, 20ml) . The solvent was removed by evaporation under reduced pressure and the residue taken up in tetrahydrofuran. 2-Iodoethanol (3.4g, 20mmol) was added dropwise. The mixture was refluxed for 24 hours then cooled, diluted with diethyl ether and 5 washed with water. After drying (MgS0 ₄ ) and evaporation, the residue was purified by chromatography on silica gel eluting with dichloromethane in hexane (1:1 to 1:0) to give p_- (2-hydroxyethoxy)bromobenzene (0.890g, 21%) .

0 This material (0.586g, 2.7mmol) in tetrahydrofuran (10ml) was treated with triethylamine (0.7ml, 5mmol) and chlorotrimethyl silane (0.63ml, 5mmol) . After 15min the mixture was diluted with ether and filtered. The filtrate

was concentrated under reduced pressure, taken up in ether, ^• refiltered, and evaporated to give the silyl ether, (0.74g) . To a solution of this aryl bromide (0.74g, 2.6mmol) in tetrahydrofuran (10ml) at -78°C under nitrogen was added a 5 solution of n-butyllithium (1.7ml, 1.5M in hexane). The mixture was stirred at -78°C for 15min, when to the resulting solution was added anhydrous cerium (III) chloride (0.74g, 2.6mmol). After a further hour at -78°C a solution of the protected monamide prepared as in Example 2b (0.84g, 0 1.4mmol) in tetrahydrofuran (8ml) was added dropwise. After stirring at -78°C for 15min acetic acid (0.15g), diethyl ether and water were added and the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure. 5

This material was taken up in tetrahydrofuran (30ml) and 0.4M hydrochloric acid (6ml) was added. After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (6ml) was added, and the mixture extracted with ethyl acetate. The 0 organic layer was dried (MgS0 ) , evaporated and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-10%) to give the title compound as a white foam (0.28g, 43%); v _maχ (KBr) 3423, 2924, 2855,and 1619cm- ¹ ; λ _maχ (EtOH) 273nm (ε _m 19,200); δ _R 5 (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 14-H ₃ ), 1.4 (IH, m, 12-H), 2.0 (IH, m, 8-H), 2.15 (3H, d, J 0.6Hz, 15-H ₃ ) , 2.3 (IH, dd, J 9.5 and 14.4Hz, 4-H), 2.7-2.85 (3H, m, 4-, 10- and 11-H), 3.4-3.9 (8H, m, 5, 6, 7, 13-H, 16-H ₂ and ethyl-2-H ₂ ) , 4.1 (2H, t, 4.9Hz, ethyl-l-H ₂ ) , 6.85

30 (IH, bs, 2-H), 7.0 and 7.95 (4H, ₂ B ₂ q, J 8.8Hz, Ar-H ₄ ); δ _c (CD3OD) 12.3 (C-17), 20.3 (C-15), 20.4 (C-14) , 33.0 (C-9) ,

41.7 (C-8), 43.7 (C-12) , 44.2 (C-4 ) , 56.9 (C-10) , 61.3 (C-ll), 61.5 (CH ₂ OH) , 66.4 (C-16) , 80.0 (C-6) , 70.7 (C-13) ,

70.8 (Ar OCH ₂ ), 71.7 (C-7) , 76.4 (C-5) , 115.4 (Ar C-3 and 355), 123.8 (C-2), 131.7 (C-2 and 6), 133.2 (ArC-1) , 157.5

(C-3), 164.2 (Ar C-4), 192.6 (C-l) ; m/z (FAB, Thioglycerol

matrix) 465 (MH ⁺ ) , 482 (MNH ₄ ⁺ ) .

Example 20

4-Methylthiomethoxyphenyl-l-normon-2-yl ketone

To 4-bromophenol (1.73g, lOmmol) in tetrahydrofuran (20ml) was added a methanol solution of tetrabutyl ammonium hydroxide (1M, 10ml) . The solvent was removed by evaporation under reduced pressure, and the solid residue taken up in THF (20ml) . Chloromethyl methyl sulphide (0.8ml, lOmmol) was added dropwise and the mixture stirred at 20°C for lh. The solvent was then removed and the residue triturated with hexane. The hexane soluble material was purified by chromatography on silica eluting with 20-40% dichloromethane in hexane to give 4- (methylthiomethoxy)bromobenzene (1.6g, 68%) .

To a solution of the above aryl bromide (0.70g, 3mmol) in tetrahydrofuran (12ml) at -78°C under nitrogen was added a solution of n-butyllithium (6.66ml, 1.5M in hexane) . The mixture was stirred at -78°C for 15min, when to the resulting solution was added anhydrous cerium (III) chloride (0.738g, 3mmol) . After a further 1 hour at -78°C a solution of the protected monamide prepared as in Example 2b (1.20, 2.0mmol) in tetrahydrofuran (10ml) was added dropwise. After stirring at -78°C for 15min acetic acid (0.20g) was added, the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure.

This material was taken up in tetrahydrofuran (40ml) and 0.4M hydrochloric acid (8ml) was added. After 2.5min at 20°C saturated aqueous sodium hydrogen carbonate (8ml) was

added, and the mixture extracted with diethyl ether. The organic layer was dried (MgS0 ₄ ) , evapora _. ted and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title 5 compound as a white foam (0.48g, 50%); v _maχ (KBr) 3429, 2922, 1655, 1599 and 1167cm- ¹ ; λ _maχ (EtOH) 293nm (E _JJJ 19,800); δ _R (CD3OD) 0.95 (3H, d, J 7.2Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, I4-H3) , 1.4 (IH, m, 12-H), 1.7 (2H, m, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.2 (3H, d, J 0.6Hz, I5-H3) , 2.25

10 (3H, s, SMe), 2.35 (IH, dd, J 9.4, 14.4Hz, 4-H), 2.7-2.9 (3H, m, 4 , 10 and 11-H), 3.4-3.9 (6H, m, 5, 6,7, 13-H and 16-H ₂ ), 5.7 (2H, s, S-CH ₂ ) , 6.85 (IH, bs, 2-H), 7.05 and 7.9 (4H, A ₂ B ₂ q, J 8.9Hz, Ar-H ₄ ) ; δ _c (CD3OD) 12.3 (C-17) , 14.5 (S-Me) , 20.3, 20.4 (C14 and 15), 33.0 (C-9) , 41.8 (C-8) ,

1543.7 (C-12), 44.2 (C-4), 56.9 (C-10) , 61.3 (C-ll) , 66.4 (C-16), 70.1, 70.7 and 71.7 (C-6, 7 and 13), 76.4 (C-5) , 116.6 (Ar C-3 and C-5), 123.7 (C-2), 131.5 (Ar-C-2 and C-6), 133.8 (Ar C-l) , 157.8 (C-3), 162.4 (Ar C-4) and 192.6 (C-l); m/z 480 (M ⁺ , 2%), 422 (2) and 181 (100). (Found: M ⁺ ,

20 480.2177. C ₂₅ H ₃₄ 0 ₇ S requires M, 480.2182) .

Example 21

4-(1-Hydroxyimino)ethylphenyl-l-normon-2-yl ketone

25

To 4-bromoacetophenone oxime (0. 3g, 2mmol) in tetrahydrofuran (5ml) was added triethylamine (0.7ml, 2mmol) and chlorotrimethyl silane (0.5ml, 4mmol) . After 45 minutes hexane was added and the mixture filtered. The volatiles

30 were removed under reduced pressure, and the residue re-dissolved in hexane, filtered and concentrated to a colourless liquid, 0.58g.

To the above material (0.45g) in tetrahydrofuran (5ml) at 35 -78°C under nitrogen was added butyllithium (1ml, 1.5M ) dropwise over Imin. After a further minute a solution of

the protected monamide prepared as in Example 2b (0.6g, lmmol) in THF (4ml) was added dropwise. -After a further 15min at -78°C acetic acid (0.09g), diethyl ether and water were added. The organic phase was washed with brine, dried and evaporated. After partial purification by chromatography on silica gel eluting with 0-15% ethyl acetate in hexane, the product in tetrahydrofuran (50ml) was treated with 0.4M HC1 (10ml) at 20°C for 2min. After quenching with saturated aqueous sodium hydrogen carbonate (10ml), the mixture was extracted with diethyl ether. The ether soluble material was purified by chromatography on silica gel to give the title compound as a white solid after crystallisation from diethyl ether/hexane (0.245g, 35%); mp 73-75°C; V _maχ (KBr) 3365, 2967, 2920, 1653, 1609 1249 . and 1004cm- ¹ ; λ _maχ (EtOH) 292nm (ε _m 21,200) and 225nm

(9,300); δ _H (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 7.4Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6. 1Hz , 9-H ₂ ), 1.95 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ) , 2.25 (3H, s, oxime Me), 2.4 (IH, dd, J 9.5, 14.4Hz, 4-H), 2.7-2.9 (3H, m, 4, 10 and 11-H), 3.4-3.9 (6H, m, 5, 6,7, 13-H and 16-H ₂ ) , 6.9 (IH, s, 2-H), 7.75 and 7.95 (4H, A ₂ B ₂ q, J 8.5Hz, Ar-H ₄ ); m/z. 461 (M ⁺ , 1%), 444 (1), 408 (1) and 162 (100) . (Found: 461.2424. C ₂₅ H ₃₅ N0 ₇ requires 461.2414) .

Example 22

5-Indolyl-l-normon-2-yl ketone

To a solution of 5-bromoindole (0.3g, 1.5mmol) in tetrahydrofuran (8ml) at 0°C under nitrogen was added sodium hydride (0.072g, 50% in oil) . After lh. the resulting suspension was cooled to -78°C and a solution of n-butyllithium (1.0ml, 1.5M in hexane) added dropwise. The mixture was stirred at -78°C for 30min, when to the resulting mixture was added dropwise a solution of the protected monamide prepared as in Example 2b (0.4g,

0.66mmol) in tetrahydrofuran (2ml). After stirring at -78°C for 15min and lOmin at 0°C, the reaction was quenched with acetic acid (0.06g), and diluted with diethyl ether and washed with water and brine. After drying (MgS0 ₄ ) and 5 evaporation under reduced pressure the tris-protected product was partially purified by chromatography on silica gel eluting with 0-6% diethyl ether in dichloromethane.

This material was dissolved in tetrahydrofuran (10ml) and

10 treated with 0.4M HC1 (2ml) at 20°C for 2min. Saturated aqueous sodium hydrogen carbonate (2ml) and diethyl ether were added, and the organic layer was washed with brine, dried (MgS0 ₄ ) and concentrated by evaporation under reduced pressure. Chromatography on silica gel eluting with 2-12%

15 methanol in dichloromethane gave the title product as a colourless foam (O.lOg, 34%); v _maχ (KBr) 3401, 2969, 2911, 1645, 1611, 1349, 1250, 1104 and 1053cm- ¹ ; λ _maχ (EtOH) 307 (ε _m 13,030) and 263nm (29,020); δ _H (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ), 1.2 (3H, d, J 6.4Hz, 14-H ₃ ) , 1.4 (IH, m,

20 12-H), 1.7 (2H, t, J 6.8Hz, 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.15 (3H, s, 15-H ₃ ), 2.4 (IH, dd, J 14.3, 9.5Hz, 4-H), 2.7 (IH, dd, J 7.7, 2.2Hz, 11-H), 2.75 (IH, d, J 14.3Hz, 4-H), 2.85 (IH, dt, J 2.2, 6.1Hz, 10H ₂ ), 3.4-4.0 (6H, m, 5, 6, 7, 13-H, 16-H ₂ ), 6.4 (IH, d, 3.3Hz, indole 3-H) , 6.95 (IH, s, 2-H),

257.3 (IH, d, J 3.3Hz, indole 2-H), 7.4 (IH, d, J 8.7Hz, indole 7-H) , 7.8 (IH, dd, J 1.7, 7.8Hz, indole 6-H) and 8.3 (IH, d, J 1.7Hz, indole 4-H); m/z.443 (M ⁺ , 5%) and 144 (100). (Found: 443.2303. C ₂₅ H ₃₃ NOg requires 443.2308) .

30 Example 23

2-Hydroxyphenyl-l-normon-2-yl ketone

To a solution of 2-bromophenol (0.233g, 2.0mmol) in 35 tetrahydrofuran (5ml) under nitrogen was added sodium hydride (0.06g, 80% in oil, 2mmol) . The resulting yellow

solution was cooled to -78°C and butyllithium(1.33ml, 1.5M in hexane) added dropwise. After a further five minutes a solution of the protected monamide prepared as in Example 2b (0.6g, lmmol) in tetrahydrofuran (5ml) was added dropwise. 5 After 40min at -78°C acetic acid (0.25g) was added, the mixture diluted with diethyl ether, and washed with water and brine. After drying (MgS0 ₄ ) and evaporation the crude tris-protected product was partially purified by chromatography on silica gel eluting with 0-20% ethyl 10 acetate in hexane.

This material was taken up in tetrahydrofuran (20ml) and 0.4M hydrochloric acid (4ml) was added. After 2min at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was added,

15 and the mixture extracted with diethyl ether. The organic layer was dried (MgS0 ₄ ) , evaporated and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title compound as a yellow foam (O.llOg, 26%); V _maχ (KBr) 3423, 2969, 2925,

20 1636, 1586, 1487, 1443, 1294, 1240 and 757cm- ¹ ; λ _maχ (EtOH) 338nm (ε _m 35,800), 276 (17,900) and 214nm (14,600); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.5Hz, 14-H ₃ ), 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.5Hz, 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ) , 2.4 (IH, dd, J 9.5,

25 14.3Hz, 4-H), 2.7-2.8 (3H, m, 4-, 10 and 11-H), 3.4-3.9 (6H, m, 5, 6, 7, 13-H, 16-H ₂ ) , 6.9 (3H, m, 2-, Ar 3- and Ar 5-H) , 7.5 (IH, dt, J 1.5, 7.1Hz, Ar 4-H) and 7.9 (IH, dd, J 1.5 and 8.2Hz, Ar 6-H) ; m/z _. 420 (M ⁺ , 1%) and 161 (100) . (Found: 420.2156. C ₂₃ H ₃₂ 0 ₇ requires 420.2148) .

30

Example 24

4-Hydroxyiminomethylphenyl-l-normon-2-yl ketone

To the 4-formyl ketone prepared as in Example 13 (0.09g, 0.20mmol) in methanol (2ml) was added sodium acetate trihydrate (0.04g) and hydroxylamine hydrochloride (0.02g). After 30min. the solvent was removed by evaporation under reduced pressure, and the residue loaded onto a silica gel column. Eluting with 4-10% methanol in dichloromethane gave the title compound as a white amorphous solid (0.071g, 79%); V _maχ (KBr) 3371, 2969, 2913, 1655, 1609 and 1247cm- ¹ ; λ _maχ (EtOH) 294nm (ε _m 24,400); δ _R (CD ₃ 0D) 0.95 (3H, d, J 7.0Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.7Hz, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.2 (3H, s,

15-H ₃ ) , 2.4 (IH, dd, J 9.5, 14.3Hz, 4-H), 2.7-2.8 (3H, m, 4-, 10 and 11-H), 3.4-3.9 (6H, m, 5, 6, 7, 13-H, 16-H ₂ ) , 6.9 (IH, s, H-2), 7.7 and 7.9 (A ₂ B ₂ q, J 8.4Hz, Ar-H ₄ ) and 8.1 (IH, s, oxime-H) ; m/z.447 (M ⁺ , 1%), 430 (1), 412 (1) and 148 (100). (Found: 447.2254. C ₂₄ H ₃₃ N0 ₇ requires 447.2257) .

Example 25

4-MethoxyiminomethylphenyI-l-normon-2-yl ketone

To the 4-formyl ketone prepared as in Example 13 (0.06g, 0.14mmol) in methanol (1ml) was added sodium acetate trihydrate (0.03g) and p_-methylhydroxylamine hydrochloride (0.015g) . After 30min. the solvent was removed by evaporation under reduced pressure, and the residue loaded onto a silica gel column. Eluting with 4-10% methanol in dichloromethane gave the title compound as a white amorphous solid (0.018g, 30%); V _maχ (KBr) 3439, 2969, 1656, 1610, 1246 and 1052cm- ¹ ; ^ _ax (EtOH) 296nm ^ 27,700); δ _R (CD3OD)

-59-

0.95 (3H, d, J 7.0Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.7Hz, .9-H ₂ ) , 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ), 2.4 (IH, dd, J 9.5, 14.3Hz, 4-H), 2.7-2.8 (3H, m, 4-, 10 and 11-H), 3.4-3.9 (6H, m, 5, 6, 7, 5 13-H, 16-H ₂ ), 3.95 (3H, s, methoxy-H ₃ ) , 6.9 (IH, s, H-2) , ^' 7.7 and 7.9 (A ₂ B ₂ q, J 8.4Hz, Ar-H ₄ ) and 8.15 (IH, s, oxime-H) ; m/z. 461 (M ⁺ , 1%) and 148 (100) . (Found: 461.2424. ^C 25 ^H 35 ^N0 7 ^rec 2 ^uires 461.2414 ⁾ .

10 Example 26

l-Normon-2-yl 4-propylthioρhenyl ketone

To 4-bromophenylpropyl sulphide (1.07g, 4.6mmol) in 15 tetrahydrofuran (20ml) at -78°C under nitrogen was added dropwise a solution of n-butyllithium in hexane (2.6ml, 1.5M). After 15min at -78°C anhydrous cerium (III) chloride (0.984g, 4mmol) was added, and the mixture stirred for a further 45min at -78°C before dropwise addition of a 20 solution of the protected monamide (1.2g, 2mmol) in tetrahydrofuran (10ml) , prepared as in Example 2b.

After 30min at -78°C, acetic acid (0.25g), diethyl ether and water were added, and the organic phase washed with brine.

25 After drying (MgS0 ₄ ) and evaporation, the residue was taken up in tetrahydrofuran (50ml) and treated with 0.4M hydrochloric acid (10ml) for 2min. The reaction was quenched with saturated sodium hydrogen carbonate (10ml) and extracted with diethyl ether. The organic extracts were

30 washed with brine, dried (MgS0 ₄ ) and evaporated. The residue was purified by chromatography on silica gel eluting with methanol in dichloromethane (0-4%) to give the title product (0.399g, 41%) as a white foam; v _maχ (KBr) 3423, 2964, 2926, 1652, 1615, 1588, 1251 and 1092cm- ¹ ; λ _maχ (EtOH)

35 321 (ε _m 19,300), 263 (9,400) and 246nm (10,000); δ _R (CD3OD)

0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.05 (2H, t, J 7.4Hz, propyl 3-H ₃ ), 1.2 (3H, d, J 6.4Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (4H, m, 9- and propyl 2-H ₂ ) , 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ) , 2.35 (IH, dd, J 9.4, 14.4Hz, 4-H), 2.7-2.8 (3H, m, 54-, 10 and 11-H), 3.0 (2H, t, J 7.2Hz, propyl 1-H ₂ ) , 3.4-3.9 (6H, m, 5, 6, 7, 13-H, 16-H ₂ ) , 6.85 (IH, s, 2-H), 7.35 and 7.9 (4H, A ₂ B ₂ q, J 8.4Hz, Ar-H ₄ ) ; m/z.478 (M ⁺ , 6%) and 179 (100). (Found: 478.2400. C ₂ gH ₃ gSOg requires 478.2389) .

10 Example 27

4-Propylsulphinylρhenyl-l-normon-2-yl ketone

To the p_-propylthiophenyl ketone from Example 26, (0.044g,

150.09mmol) in dichloromethane (2ml) was added saturated aqueous sodium hydrogen carbonate (1ml) . The mixture was cooled to 0°C and m-chloroperbenzoic acid (0.018g) added. After stirring at 0°C for 15min the mixture was diluted with dichloromethane (50ml) , the organic layer dried (MgS0 ₄ ) and

20 evaporated. The residue was purified by chromatography on silica eluting with methanol in dichloromethane (0-8%) . The title product was obtained as a white foam, (0.033g, 72%); V _maχ (KBr) 3430, 2968, 2929, 1657, 1608, 1241 and 1006cm- ¹ ; λ _maχ (EtOH) 277nm (ε _m 19,200) and 203nm (22,700); δ _R (CD ₃ OD)

25 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.05 (3H, t, J 7.4Hz,

CH ₂ CH ₂ CH ₃ ), 1.2 (3H, d, J 6.5Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.6-1.9 (4H, m, CH ₂ CH ₂ CH ₃ and 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ), 2.4 (IH, dd, J 9.6, 14.3Hz, 4-H), 2.6-3.0 (5H, m, CH ₂ CH ₂ CH ₃ , 4, 10 and 11-H), 3.4-4.0 (6H, m, 5, 6, 7,

30 13-H, 16-H ₂ ), 6.9 (IH, s, 2-H), 7.8 and 8.1 (4H, A ₂ B ₂ q, J 8.3Hz, Ar-H ₄ ); m/z _. (FAB, 3-NOBA/Na) , 517 (MNa ⁺ ) .

Example 28

4-Propylsulphonylphenyl-l-normon-2-yl ketone

To the p_-propylthiophenyl ketone from Example 26, (0.044g, 0.09mmol) in dichloromethane (2ml) was added saturated aqueous sodium hydrogen carbonate (1ml) . The mixture was cooled to 0°C and m-chloroperbenzoic acid (0.036g) added. After stirring at 0°C for 90min the mixture was diluted with dichloromethane (50ml) and the organic layer dried (MgS0 ₄ ) and evaporated. The residue was purified by chromatography on silica eluting with methanol in dichloromethane (0-8%) . The title product was obtained as a white foam, (0.027g, 57%); v _maχ (KBr) 3446, 2969, 2925, 1661, 1609 and 1147cm- ¹ ; λ _maχ (EtOH) 270 (ε _m 14,800) and 252nm (15,900); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.0 (3H, t, J 7.4Hz, CH ₂ CH ₂ CH ₃ ) , 1.2 (3H, d, J 6.5Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (4H, m, CH ₂ CH ₂ CH ₃ and 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.25 (3H, s, 15-H ₃ ), 2.4 (IH, dd, J 9.6, 14.3Hz, 4-H), 2.75 (3H, m, 4, 10 and 11-H), 3.2 (2H, m, CH ₂ CH ₂ CH ₃ ) , 3.4-4.0 (6H, , 5, 6, 7, 13-H, 16-H ₂ ), 6.9 (IH, s, 2-H), 8.0 and 8.15 (4H, A ₂ B ₂ q, J 8.4Hz, Ar-H ₄ ); m/z.510 (M ⁺ , 1%) and 211 (100) . (Found: 510.2285. C ₂ gH ₃ gOgS requires 510.2287) .

Example 29

4-Methylsulphinylmethoxyphenyl-l-normon-2-yl ketone

To the 4-methylthiomethoxyphenyl ketone, prepared as in Example 20, (0.065g, 0.135mmol) in dichloromethane/ saturated aqueous sodium hydrogen carbonate (2ml:1ml) at 0°C was added m-chloroperbenzoic acid (0.025g) . The mixture was stirred at 0°C for 15min before the mixture was diluted with dichloromethane, the organic layer dried and purified by

chromatography on silica eluting with 0-8% methanol in dichloromethane to give the title compound as a white foam (0.025g, 37%); v _maχ (KBr) 3423, 2968, 2921, 1655, 1600, 1244, 1171 and 1030cm- ¹ ; λ _maχ (EtOH) 284nm (ε _m 19,100); δ _R 5 (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.5Hz, 14-H ₃ ), 1.4 (IH, m, 12-H), 1.7 (2H, m, 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.2 (3H, d, J 0.6Hz, 15-H ₃ ) , 2.35 (IH, dd, J 9.6, 14.4Hz, 4-H), 2.7 (3H, s, SMe), 2.7-2.8 (3H, m, 4, 10 and 11-H), 3.4-3.9 (6H, m, 5, 6, 7, 13-H, 16-H ₂ ) , 5.15 and 5.13 0 (2H, ABq, J 10.7Hz, SCH ₂ ) , 6.85 (IH, bs, 2-H), 7.2 and 8.0 (4H, A ₂ B ₂ q, J 8.8Hz, Ar-H ₄ ) ; m/z (FAB, NOBA/Na) 519 (MNa ⁺ ) .

Example 30

154-Methylsulphonylmethoxyphenyl-l-normon-2-yl ketone

To the 4-methylthiomethoxyphenyl ketone, prepared as in Example 20, (0.075g, 0.156mmol) in dichloromethane/ saturated aqueous sodium hydrogen carbonate (2ml:1ml) at 0°C

20 was added m-chloroperbenzoic acid (0.055g) . After stirring at 0°C for 2h the layers were separated, the aqueous extracted with dichloromethane and the combined organic layers dried, concentrated and purified by chromatography on silica eluting with 0-8% methanol in dichloromethane to give

25 the title compound as a white foam (0.034g, 42%); ^v _maχ

(KBr) 3437, 2970, 2926, 1655, 1601, 1319, 1244 and 1137cm- ¹ ; λ _maχ (EtOH) 276nm (ε _m 18,500); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ), 1.2 (3H, d, J 6.5Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, m, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.2 (3H, s,

30 15-H ₃ ), 2.35 (IH, dd, J 9.6, 14.4Hz, 4-H), 2.7 (IH, dd, J 2.1, 7.5Hz, 11-H), 2.8 (2H, m, 4 and 10-H), 3.0 (3H, s, S-Me) , 3.4-3.9 (6H, m, 5, 6, 7, 13-H, 16-H ₂ ) , 5.3 (2H, s, S-CH ₂ ), 6.85 (IH, s, 2-H), 7.2 and 8.0 (4H, ₂ B ₂ q, J 8.9Hz, Ar-H ₄ ); m/z.512 (M ⁺ , 0.02%) and 213 (100). (Found: M,

35512.2064. C ₂₅ H ₃₆ 0 ₉ S requires M, 512.2080).

Example 31 p-Aminophenyl-l-normon-2-yl ketone

p-Bromoaniline (5g, 29mmol) was treated with N,N- 5 diethyltrimethylsilylamine (24ml) and a catalytic amount of ammonium sulphate at reflux for 12h. Distillation gave N,N-bistrimethylsilyl-p-bromo aniline (3.8g, 41%) bp 130-137°C (7mmHg) .

10 To the above material (0.35g, 1.lmmol) in tetrahydrofuran (4ml) at -78°C under nitrogen was added dropwise butyllithium (0.7ml, 1.5M) . After 15min. a solution of the protected monamide prepared as in Example 2b (0.4g, 0.67mmol) in THF (4ml) was added dropwise. After a further

15 15min. at -78°C acetic acid (0.06g), diethyl ether and wate were added. The organic phase was washed with brine, dried and evaporated. Partial purification by chromatography on silica gel eluting with 0-15% ethyl acetate in hexane gave the N,N, 6, 7, 13 o-pentatrimethylsilvl derivative of the

20 title product, (0.088g, 17%) . A portion of this material (0.060g) in tetrahydrofuran (10ml) was treated with 0.4M HC (2ml) at 20°C for 2min. After quenching with saturated aqueous sodium hydrogen carbonate (2ml) , and extraction wit diethyl ether, the ether soluble material was purified by

25 chromatography on silica gel to give the title compound as a colourless foam, (0.027g, 10%); V _maχ (KBr) 3356, 3236, 2970, 2903, 1634, 1592, 1259 and 1173cm- ¹ ; λ _maχ (EtOH) 337 (ε _m 18,100) and 243.5nm (11,400); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 14-H ₃ ) , 1.4 (IH, m,

30 12-H), 1.7 (2H, m, 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.1 (3H, S, 15-H ₃ ) , 2.3 (IH, dd, J 9.4 and 14.4Hz, 4-H), 2.7 (2H, m, 4 and 11-H), 2.8 (IH, dt, J 2.4 and 6.0Hz, 10-H), 3.4 (IH, dd, J 3.1 and 8.8Hz, 6-H) , 3.6 (IH, dd, J 16 and 11.5Hz, 16-H), 3.7-3.9 (4H, m, 5, 7, 13 and 16-H), 6.6 and 7.7 (4H, A ₂ B ₂ q J

35 8.7Hz, Ar-H), 6.7 (IH, s, 2-H); m/z.419 (M ⁺ , 2%), 120 (100) . (Found: M ⁺ , 419.2319. C ₂₃ H ₃₃ NOg requires 419.2303).

Example 32

l-Normon-2-yl-p-ureidophenyl ketone

The reaction of Example 31 was carried out on 1.5 times the scale to give the crude fully silylated ketone (0.91g) . A portion (0.77g) was taken in THF/water (5ml + 1ml) and acetic acid (O.lg) added. After 5min. at 20°C saturated aqueous sodium hydrogen carbonate (2ml) was added, and the mixture extracted with diethyl ether. The organic layer was washed with brine, dried (MgS0 ₄ ) and evaporated. The residue was taken up in tetrahydrofuran (2ml) and treated with excess trichloroacetyl isocyanate. After lOmin. at 20°C water and diethyl ether were added, the organic layer dried and evaporated, and the residue purified by chromatography on silica gel eluting with 10-30% ether in hexane to give 6,7,13-o-tristrimethylsilyl-l-normon-2- yl-p-N'-trichloroacetylureidophenyl ketone (0.095g, 14% overall) . This was taken up in THF (5ml) and treated with 0.4M HC1 (1ml) at 20°C for 2min. The reaction was quenched with saturated sodium hydrogen carbonate and extracted with ethyl acetate. The solvent was removed and the residue treated with methanol at 20°C for 3h. After evaporation the crude product was purified by chromatography eluting with 5-15% methanol in dichloromethane to give the title ketone (35mg, 65%); V _maχ (KBr) 3358, 2970, 2923, 1677, 1585, 1528 and ¹² 51cm_l. ^^ (EtOH) 308 (ε _m 19,800) and 232nm (9,300); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.5Hz, 14-H ₃ ), 1.4 (IH, m, 12-H), 1.7 (2H, m, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ) , 2.35 (IH, dd, J 9.5 and 14.4Hz, 4-H), 2.7-2.8 (3H, m, 4, 10 and 11-H), 3.4 (IH, dd, J 3.0 and 8.9Hz, 6-H) , 3.6 (IH, d, J 11.4Hz, 16-H), 3.8-3.9 (4H, m, 5, 7, 13 and 16-H) 6. (IH, s, 2-H), 7.5 and 7.9 (4H, A ₂ B ₂ q, J 8.8Hz, ArH) ; m/z (FAB, 3-Noba/Na matrix) 485 (MNa ⁺ ) .

Example 33

p-Ethylaminophenyl-l-normon-2-yl ketone

To a solution of p_-bromo N-ethyl-N-trimethylsilyl aniline (0.544g, 2mmol) (prepared from p_-bromo N-ethyl aniline, N,N-diethyl trimethylsilylamine and a catalytic amount of ammonium sulphate) in tetrahydrofuran (8ml) at -95°C under nitrogen was added a solution of ri-butyllithium (1.0ml, 1.5M in hexane) . The mixture was stirred at -95°C for lOmin, when to the resulting solution was added dropwise a solution of the protected monamide prepared as in Example 2b (0.6g, l.Ommol) in tetrahydrofuran (5ml) . After stirring at -90 to -78°C for 15min. acetic acid (0.12) diethyl ether and water were added and the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure to a pale yellow/green oil. Preliminary purification by chromatography on silica gel eluting with 10-15% ethyl acetate in hexane gave the crude tris-protected product (O.llg) which was taken up in tetrahydrofuran (20ml) and 0.4M hydrochloric acid (4ml) was added. After 2.5min. at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was added, and the mixture extracted with diethyl ether. The organic layer was dried (MgS0 ₄ ), evaporated, and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title compound as a yellow/green foam (0.07g, 16%); ^v _maχ (KBr) 3365, 3055, 2970, 2874, 1646, 1609, 1260 and 1174cm- ¹ ; λ _mav (EtOH) 350 (ε _m 22,100) and 246nm (11,800); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.0Hz, 17-H ₃ ), 1.2 (3H, d, J 6.5Hz, 14-H ₃ ) , 1.25 (3H, t, J 7.1Hz, Et-2-H ₃ ), 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.5Hz, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.1 (3H, s, 15-H ₃ ) , 2.3 (IH, dd, J 9.4 and 14.4Hz, 4-H), 2.7 (2H, m, 4- and 11-H), 2.8 (IH, dt, J 1.3 and 5.8Hz, 10-H), 3.2 (2H, q, J 7.1Hz, Et-l-H ₂ ), 3.4-3.9 (6H, m, 5, 6, 7, 13-H and 16-H ₂ ), 6.7 (IH,

s, 2-H), 6.6 and 7.8 (4H, A ₂ B ₂ q, J 8.9Hz, Ar-H ₄ ) ; m/z.447 (M+, 5%), 148 (100). (Found: 447.2640. C ₂₅ H _3? NOg requires 447.2621) .

Example 34

l-Norm-2-yl-p-propanamidophenyl ketone

The reaction of Example 31 was carried out on 0.72 x the scale and the crude product treated as in Example 32 to give the 6, 7, 13-o-tris-protected ketone. This material was reacted with propionic anhydride (0.2ml) in the presence of 2,6-lutidine (0.3ml) and a catalytic amount of 4-DMAP using tetrahydrofuran (5ml) as solvent. After 6h at 20°C diethyl ether and water were added, and the organic phase washed with 5% citric acid, water, saturated aqueous sodium hydrogen carbonate and brine. Partial purification by chromatography on silica gel eluting with 0-30% ethyl acetate in hexane gave the tris protected title ketone (lOOmg) . This material was taken up in tetrahydrofuran (10ml) and 0.4M hydrochloric acid (2ml) was added. After 2.5min. at 20°C saturated aqueous sodium hydrogen carbonate (2ml) was added, and the mixture extracted with diethyl ether. The organic layer was dried (MgS0 ₄ ), evaporated, an the residue purified by chromatography on silica gel elutin with methanol in dichloromethane (2-8%) to give the title compound as a colourless foam (0.048g, 14% overall); v.„_ _v (KBr) 3425 and 1247cm- ¹ ; λ _maχ (EtOH) 303 (ε _m 23,600) and 228nm (10,600); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (6H, m, 14-H ₃ ) and propionate 3-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, m, 9-H) , 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ) , 2.4 (3H, m, 4-H and propionate 2-H ₂ ) , 2.7 (IH, dd, J 2.0 and 7.6Hz, 11-H), 2.8 (2H, m, 4- and 10-H), 3.4 to 3.9 (6H, m, 5-, 6-, 7- and 13-H and 16-H ₂ ) , 6.9 (IH, s, 2-H), 7.7 and 7.9 (4H,

A ₂ B ₂ q, J 8.7Hz, Ar-H ₄ ) ; m/z. 475 (M ⁺ , 1%), 457 (1) and 176 (100). (Found: 475.2575. C ₂ gH ₃₇ N0 ₇ requires 475.2570) .

Example 35

p-Imidazol-4 (5) -yl phenyl-l-normon-2-yl ketone

4 (5) -p_-bromophenyl imidazole was prepared using the method of Chem. Ber. 1960, 2083. To a solution of this material (0.335g, 1.5mmol) in tetrahydrofuran (5ml) at ^0o C under nitrogen was added sodium hydride (0.043g, 80% in oil) . After lh. the resulting suspension was cooled to -78°C and a solution of n-butyllithium (1.0ml, 1.5M in hexane) added dropwise. The mixture was stirred at -78°C for 30min, when to the resulting mixture was added dropwise a solution of the protected monamide prepared as in Example 2b (0.9g, 1.5mmol) in tetrahydrofuran (5ml) . After stirring at -78°C for 30min. the reaction was quenched with acetic acid (0.18g), diluted with diethyl ether and washed with water and brine. After drying (MgS0 ₄ ) and evaporation under reduced pressure the tris-protected product waspartially purified by chromatography on silica gel eluting with 0-6% diethyl ether in dichloromethane.

This material was dissolved in tetrahydrofuran (10ml) and treated with 0.4M HC1 (3ml) at 20°C for 2min. Saturated aqueous sodium hydrogen carbonate (2ml) and diethyl ether were added, and the organic layer was washed with brine, dried (MgS0 ₄ ) and concentrated by evaporation under reduced pressure. Chromatography on silica gel eluting with 2-12% methanol in dichloromethane gave the title product as a colourless foam, (0.046g, 7%); v _maχ (KBr) 3404, 2969, 2925, 1649, 1609, 1250, 1108, and 1054cm- ¹ ; λ _maχ (EtOH) 319nm (ε _m 21,700); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2

(3H, d, J 6.2Hz, 14-H ₃ ), 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.5Hz, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ) , 2.4 (IH, dd, J 9.5 and 14.4Hz, 4-H), 2.7-2.8 (3H, , 4-, 10-, and 11-H), 3.4-3.9 (6H, m, 5-, 6-, 7-, and 13-H and 16-H ₂ ) , 6.9 (IH, s, 2-H), 7.6 (IH, bs, imidazole-5 (4)-H) , 7.8 (IH, d, J 0.7Hz, imidazole-2-H) , 7.85 and 8.0 (4H, A ₂ B ₂ q J 8.5Hz, Ar-H ₄ ) ; m/z. (FAB, 3 NOBA/Na) 493 (MNa ⁺ ) .

Example 36

l-Normon-2-yl-p-pyrimidin-4-yl phenyl ketone

4-p_-Bromophenyl pyrimidine was prepared by treating p_- bromoacetophenone with dimethyl sulphate and formamide followed by p_-toluene sulphonic acid (Chem Ber 1960, 1405) . To this material (0.47g, 2.0mmol) in tetrahydrofuran (5ml) at 95°C under nitrogen was added dropwise a solution of n-butyllithium (1.0ml, 1.5M in hexane) . After lmin. to the resulting brown solution was added dropwise a solution of the protected monamide prepared as in Example 2b (0.6g, l.Ommol) in tetrahydrofuran (2ml) . After stirring at -95 to -50°C for 20min. the reaction was quenched with acetic acid (0.13g), diluted with diethyl ether and washed with water and brine. After drying (MgS0 ₄ ) and evaporation under reduced pressure the tris-protected product was partially purified by chromatography on silica gel eluting with 10-25% ethyl acetate in hexane.

This material was dissolved in tetrahydrofuran (10ml) and treated with 0.4M HC1 (2ml) at 20°C for 2min. Saturated aqueous sodium hydrogen carbonate (2ml) and diethyl ether were added, and the organic layer was washed with brine, dried (MgS0 ₄ ) and concentrated by evaporation under reduced pressure. Chromatography on silica gel eluting with 0-12% methanol in dichloromethane gave the title product as a colourless foam, (0.063g, 13%); v _maχ (CH ₂ C1 ₂ ) 3600, 2935,

-69-

1660, 1605, 1580 and 1110cm- ¹ ; λ _maχ (EtOH) 289.5nm (ε _m 16,500); δ _R (CD3OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, I4-H3), 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.4Hz, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.25 (3H, s, 15-H ₃ ) , 2.4 (IH, dd, J 9.5 and 14.3Hz, 4-H), 2.7 (IH, dd, J 2.1 and 7.3Hz, 11-H), 2.8 (2H, m, 4- and 10-H), 3.4-3.9 (6H, m, 5-, 6-, 7-, and 13-H and 16-H ₂ ) , 6.95 (IH, s, 2-H), 8.0 (IH, dd, J 1 and 5.4Hz, pyrimidine5-H) , 8.1 and 8.3 (4H, A ₂ B ₂ q, J 8.4Hz, Ar-H ₄ ), 8.85 (IH, d, J 5.4Hz, pyrimidine-6-H) and 9. (IH, d, J 1Hz, pyrimidine-2-H) ; m/z. 482 (M ⁺ , 6%), 438 (5) and 183 (100) . (Found: 482.2433. C ₂₇ H ₃ N ₂ 0g requires 482.2417) .

Example 37

p-Ethynylphenyl l-normon-2-yl ketone

2' ,2' , 4-Tribromostyrene was prepared by the method of Corey (Tetrahedron Let 1972, 3769) and a solution of this compoun (0.34g, l.Ommol) in THF (5ml) cooled to -78°C under nitrogen. Butyllithiu (2.0ml, 1.5M in hexane) was added dropwise. After 30min. at -78°C the yellow solution was warmed to 0°C for lOmin., when a white precipitate formed. The mixture was recooled to -78°C and a solution of the protected monamide prepared as in Example 2b (0.6g, l.Ommol) in tetrahydrofuran (5ml) . After stirring at -78°C for 15min. the reaction was quenched with acetic acid (0.06g), diluted with diethyl ether and washed with water and brine. After drying (MgS0 ) and evaporation under reduced pressure the tris-protected product was partially purified by chromatography on silica gel eluting with 0-7% ethyl acetate in hexane.

This material was dissolved in tetrahydrofuran (10ml) and treated with 0. M HC1 (2ml) at 20°C for 2min. Saturated aqueous sodium hydrogen carbonate (2ml) and diethyl ether

were added, and the organic layer was washed with brine, dried (MgS0 ₄ ) and concentrated by evaporation under reduced pressure. Chromatography on silica gel eluting with 0-6% methanol in dichloromethane gave the title product as a colourless foam, (0.057g, 13%); v _maχ (KBr) 3426, 2967, 2924, 2327, 1654, 1607 and 1247cm- ¹ ; λ _maχ (EtOH) 282nm (^ 25,300); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.3Hz, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ) , 2.4 (IH, dd, J 9.4 and 14.3Hz, 4-H), 2.7-2.8 (3H, m, 4-, 10- and 11-H), 3.3 (IH, s, ethynyl-H) , 3.4-3.9 (6H, m, 5-, 6-, 7-, and 13-H and 16-H ₂ ) , 6.9 (IH, s, 2-H), 7.6 and 7.9 (4H, A ₂ B ₂ q, J 8.4Hz, Ar-H ₄ ) ; m/z (FAB, 3 NOBA/Na) 451 (MNa ⁺ ) .

Example 38

l-Normon-2-yl p-trifluoromethylphenyl ketone

To ρ_-Bromobenzotrifluoride (0.337g, l.Smmol) in tetrahydrofuran (5ml) at -78°C under nitrogen was added a solution of n_-butyllithium (1.0ml, 1.5M in hexane) . The mixture was stirred at -78°C for 5min. when to the resulting solution was added dropwise a solution of the protected monamide prepared as in Example 2b (0.6g, l.Ommol in tetrahydrofuran (5ml) . After stirring at -78°C for 15min., acetic acid (O.lOg) diethyl ether and water were added, the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure to a yellow oil.

This material was taken up in tetrahydrofuran (20ml) and 0.4M Hydrochloric acid (4ml) was added. After 2.5min. at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was added, and the mixture extracted with diethyl ether. The

organic layer was dried (MgS0 ₄ ) , evaporated and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title compound as a colourless foam (0.27g, 57%); v _maχ (KBr) 3438, 52972, 2924, 1661, 1607, 1326, 1129 and 1067cm- ¹ ; λ _maχ (EtOH) 266 (E _JJJ 14,100); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.5Hz, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.25 (3H, s, 15-H ₃ ) , 2.4 (IH, dd, J 9.3 and 14.3Hz, 4-H), 2.7-2.85 (3H, m, 4-, 10- 0 and 11-H), 3.4-3.9 (6H, m, 5-, 6-, 7- and 13-H and 16-H ₂ ) , 3.95 (3H, s, methoxy-H ₃ ) , 6.9 (IH, s, 2-H), 7.7 and 8.1 (4H, A ₂ B ₂ q, J 8.3Hz, Ar-H ₄ ); m/z. 472 (M ⁺ , 0.1%), 454 (1) and 173 (100). (Found: 472.2077. ₂₄ H ₃₁ 0gF requires 472.2073) .

Example 39

p-Ethenylphenyl l-normon-2-yl ketone

To p.-bromostyrene (0.366g, 2mmol) in tetrahydrofuran (5ml) at -78°C under nitrogen was added a solution of n.-butylllithium (1.0ml, 1.5M in hexane). The mixture was stirred at -78°C for lOmin., when to the resulting pale yellow suspension was added dropwise a solution of the protected monamide prepared as in Example 2b (0.6g, l.Ommol) in tetrahydrofuran (5ml) . After stirring at -78°C for 15min. acetic acid (O.lOg), diethyl ether and water were added, the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced pressure to a yellow oil.

This material was taken up in tetrahydrofuran (20ml) and 0.4M Hydrochloric acid (4ml) was added. After 2.5min. at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was

-72- added, and the mixture extracted with diethyl ether. The organic layer was dried (MgS0 ₄ ) , evaporated and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-4%) to give the title 5 compound as a colourless foam (0.218g, 50%); v _maχ (KBr) 3429, 3085, 2969, 2921, 1653, 1608, 1556 and 1251cm- ¹ ; λ _maχ (EtOH) 29lnm ^ 23,900); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ), 1.2 (3H, d, J 6.4Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.5Hz, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.2 (3H, s,

10 15-H ₃ ), 2.4 (IH, dd, J 9.7 and 14.4Hz, 4-H), 2.7-2.85 (3H, m, 4-, 10- and 11-H), 3.4-3.9 (6H, m, 5-, 6-, 7-, and 13-H and 16-H ₂ ), 5.4 (IH, d, J 11.2Hz, ethenyl-2-H) , 5.9 (IH, d, J 17.6Hz, ethenyl-2-H) , 6.8 (IH, dd, J 11.2 and 17.6Hz, ethenyl-1-H) , 6.9 (IH, s, 2-H), 7.75 and 7.9 (4H, ₂ B ₂ q, J

158.2Hz, Ar-H ₄ ); m/z.430 (M ⁺ , 100) . m/z.431 (MH ⁺ , 70%), 430 (M ⁺ , 100). (Found: 430.2360. ₂₅ H ₃₄ 0g requires 430.2355).

Example 40

20 p-l-Methylimidazol-4-yl phenyl-l-normon-2-yl ketone

4 (5)-p_-Bromophenyl imidazole (0.6g, 2.7mmol) was treated in tetrahydrofuran (10ml) with sodium hydride (0.081g, 80% dispersion in oil) . The mixture was cooled to 0°C and

25 methyl iodide (0.2ml) added. After 1.5h at 0°C the solvent was removed by evaporation under reduced pressure and the crude product purified by chromatography on silica gel eluting with ethyl acetate to give 1-methyl-4-p-bromophenyl imidazole (0.42g, 66%); δ _R (CDC1 ₃ ) 3.7 (3H, s, methyl-H ₃ ) ,

307.2 (IH, d, J 1Hz, imidazole-5-H) , 7.5, 7.65 (4H, A ₂ B ₂ q, J 8.6Hz, phenyl-H ₄ ) and 7.55 (IH, d, J 1Hz, imidazole- 2-H) . Later fractions contained l-methyl-5-ρ_-bromophenyl imidazole (0.14g, 22%); δ _R (CDCI3) 3.7 (3H, s, methyl-H ₃ ) ,

7.1 (IH, s, imidazole-5-H) , 7.3 and 7.6 (4H, A ₂ B ₂ q, J 8.4Hz, phenyl-H ₄ ) and 7.65 (IH, s, imidazole-2-H) . To the major product (0.412g, 1.74mmol) in tetrahydrofuran (6ml) at -100°C under nitrogen was added dropwise a solution of n-butyllithium (1.0ml, 1.5M in hexane) . The mixture was stirred at -100°C for 2min. when to the resulting mixture was added dropwise a solution of the protected monamide prepared as in Example 2b (0.9g, 1.5mmol) in tetrahydrofuran (5ml) . The reaction mixture was warmed to -78°C and stirred at this temperature for 30min. then quenched with acetic acid (0.12g), diluted with diethyl ether and washed with water and brine. After drying (MgS0 ₄ ) and evaporation under reduced pressure the tris-protected product was partially purified by chromatography on silica gel eluting with 0-6% diethyl ether in dichloromethane.

This material was dissolved in tetrahydrofuran (10ml) and treated with 0.4M HC1 (3ml) at 20°C for 2min. Saturated aqueous sodium hydrogen carbonate (2ml) and diethyl ether were added, and the organic layer was washed with brine, dried (MgS0 ₄ ) and concentrated by evaporation under reduced pressure. Chromatography on silica gel eluting with 2-12% methanol in dichloromethane gave the title product as a colourless foam, (0.128g); V _maχ (KBr) 3418, 2968, 2923, 1653, 1608, 1249, 1111 and 1050cm- ¹ ; λ _maχ (EtOH) 319nm (ε _m 22,000); δ _R (CD3OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.2Hz, I4-H3), 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.5Hz, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ) , 2.4 (IH, dd, J 9.5 and 14.5Hz, 4-H), 2.7-2.8 (3H, m, 4-, 10-, and 11-H), 3.4-3.9 (6H, m, 5-, 6-, 7-, and 13-H and 16-H ₂ ) , 3.8 (3H, s, methyl-H ₃ ) , 6.9 (IH, s, 2-H), 7.6 (IH, bs, imidazole-5-H) , 7.7 (IH, s, imidazole-

2-H) , 7.8 and 7.95 (4H, A ₂ B ₂ q J 8.5Hz, Ar-H ₄ ); m/z. 484 (M ⁺ , 3%), 185 (100) . (Found: 484.2576. C ₂₇ H ₃ g ₂ 0g requires 484.2573) .

Example 41

p-1-Methyl imidazol-5-yl phenyl-l-normon-2-yl ketone

To l-methyl-5-p.-bromophenyl imidazole (0.119g, 0.5mmol)

(prepared in Example 40) in tetrahydrofuran (2ml) at -100°C under nitrogen was added dropwise a solution of n-butyllithium (1.0ml, 1.5M in hexane) . The red solution was stirred at -100°C for 2min, when a solution of the protected monamide prepared as in Example 2b (0.3g, 0.5mmol) in tetrahydrofuran (5ml) wasadded dropwise. After warming to -78°C and stirring at -78°C for 30min. the reaction was quenched with acetic acid (0.04g), diluted with diethyl ether and washed with water and brine. After drying (MgS0 ₄ ) and evaporation under reduced pressure this material was dissolved in tetrahydrofuran (10ml) and treated with 0.4M HC1 (3ml) at 20°C for 2min. Saturated aqueous sodium hydrogen carbonate (2ml) and diethyl ether were added, and the organic layer was washed with brine, dried (MgS0 ₄ ) and concentrated by evaporation under reduced pressure.

Chromatography on silica gel eluting with 2-12% methanol in dichloromethane gave the title product as a colourless foam, (O.Olg, 3%); V' _maχ (CH ₂ C1 ₂ ) 3600, 3550, 2910, 1655, 1610 and 1110cm- ¹ ; λ _maχ (EtOH) 307nm (ε _m 15,700); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ), 1.2 (3H, d, J 6.2Hz, 14-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.5Hz, 9-H ₂ ) , 2.0 (IH, m, 8-H), 2.2 (3H, s, 15-H ₃ ), 2.4 (IH, dd, J 9.5 and 14.4Hz, 4-H), 2.7-2.8 (3H, m, 4-, 10-, and 11-H), 3.4-3.9 (6H, m, 5-, 6-, 7-, and 13-H and 16-H ₂ ) , 3.8 (3H, s, methyl-H ₃ ) , 6.9 (2H, s, 2-H) , 7.15 (IH, bs, imidazole-4-H) , 7.75 (IH, bs, imidazole-2-H) , 7.6 and 8.05 (4H, A ₂ B ₂ q, J 8.3Hz, Ar-H ₄ ) ; m/z.484 (M ⁺ , 10%), 185 (100) . (Found: 484.2576. C ₂₇ H ₃ g ₂ 0g requires 484.2573).

Example 42

p-Ethylphenyl l-normon-2-yl ketone

5 To p-bromoethylbenzene (0.278g, 1.5mmol) in tetrahydrofuran (5ml) at -78°C under nitrogen was added a solution of n-butyllithium (1.0ml, 1.5M in hexane) . The mixture was stirred at -78°C for lOmin. when to the resulting pale yellow suspension was added dropwise a solution of the

10 protected monamide prepared as in Example 2b (0.6g, l.Ommol) in tetrahydrofuran (5ml) . After stirring at -78°C for 25min. acetic acid (O.lOg), diethyl ether and water were added, the organic layer was washed with saturated aqueous sodium chloride, dried (MgS0 ₄ ) and evaporated under reduced

15 pressure to a yellow oil.

This material was taken up in tetrahydrofuran (20ml) and 0.4M Hydrochloric acid (4ml) was added. After 2min. at 20°C saturated aqueous sodium hydrogen carbonate (5ml) was added,

20 and the mixture extracted with diethyl ether. The organic layer was dried (MgS0 ₄ ) , evaporated and the residue purified by chromatography on silica gel eluting with methanol in dichloromethane (0-6%) to give the title compound as a colourless foam (0.12g, 28%); v _maχ (KBr) 3422, 2968, 1655,

25 1609, 1253 and 1052cm- ¹ ; λ _maχ (EtOH) 270nm (ε _m 20,300); δ _R (CD ₃ OD) 0.95 (3H, d, J 7.1Hz, 17-H ₃ ) , 1.2 (3H, d, J 6.4Hz, 14-H ₃ ), 1.25 (3H, t, J 7.7Hz, ethyl-2-H ₃ ) , 1.4 (IH, m, 12-H), 1.7 (2H, t, J 6.4Hz, 9-H ₂ ), 2.0 (IH, m, 8-H), 2.2 (3H, d, J 0.6Hz, 15-H ₃ ), 2.35 (IH, dd, J 9.4 and 14.4Hz,

30 4-H), 2.7-2.8 (5H, m, 4-, 10- and 11-H and ethyl-l-H ₂ ) , 3.4 (IH, dd, J 3.0 and 9.0Hz, 6-H) , 3.6 (IH, bd, J 11.6Hz, 16-H), 3.75-3.9 (4H, m, 5-, 7-, 13- and 16-H), 6.85 (IH, bs, 2-H), 7.3 and 7.85 (4H, A ₂ B ₂ q, J 8.2Hz, Ar-H ₄ ); m/z. 432 (M ⁺ , 1%), 200 (30) and 133 (100) . (Found: M ⁺ 432.2507. C ₂₅ H ₃ gOg 5 requires 432.2512) .

Bioloqical Data

The activity of the normonyl derivatives of Examples 1-42 against various bacteria* which are important in the diseases of humans was assayed in vitro using serial dilutions in nutrient agar with 5% chocolated horse blood.

The MIC values were determined after incubation for 18h at 37° and were found to be in the range 0.03 to 32μg/ml.

* H. influenzae Ql, B. catarrhalis 1502, Strep, pyogenes CN10, Strep, pneumoniae PU7 and Staph. aureus Oxford.