GB2415373A - Porphyrins for sonodynamic therapy - Google Patents

Abstract

Use of porphyrin compounds of formula I, or a metallated derivative thereof, in the preparation of a medicament for sonodynamic therapy (SDT): <EMI ID=1.1 HE=38 WI=49 LX=440 LY=698 TI=CF> <PC>wherein X1-X4 are hydrogen, a lipophilic moiety, a phenyl group, a lower alkyl, alkaryl or aralkyl group, or a cationic group of formula -L-R1-N<+>(R2)(R3)R4; L is a linker or is absent; R1 is lower alkylene, lower alkenylene, lower alkynylene, which is optionally substituted by alkyl, alkylene (optionally interrupted with oxygen), fluoro, OR5, C(O)R6, C(O)OR7, C(O)NR8R9, NR10R11 and N<+>R12R13R14; R2-R4 are hydrogen, aryl, optionally substituted lower alkyl, alkenyl or alkynyl; Z is CH or N; Y1-Y4 are absent or are aryl,lower alkyl, lower alkenyl, lower alkynyl, the latter three of which are optionally substituted by alkyl, alkylene (optionally interrupted with oxygen), aryl, OR5, C(O)R6, C(O)OR7, C(O)NR8R9, NR10R11 and N+R12R13R14; R5-R14 are hydrogen or lower alkyl; provided that at least one of X1-X4 is a cationic group as defined above and one of X1-X4 is hydrogen. Preferred metal ions for use with these porphyrins are Zn(II), Cu(II), La(III), Lu(III), Y(III), In(III), Cd(II), Mg(II), Al(III), Ru, Ni(II), Mn(III), Fe(III) and Pd(II). Metalloids including silicon and germanium may also be utilised. This therapy may be useful for killing or attenuating the growth of microorganisms, particularly bacteria, mycoplasmas, yeast, fungi and viruses, or treating dermatological infections, ulcers, infections of the lungs and cancer. An in vitro method of killing microorganisms utilising these compounds and ultrasound is also outlined.

Description

24 1 5373

NOVEL USES

Field

The present invention relates to new uses of porphyrin compounds and, in o particular, the use of such compounds as agents for sonodynamic therapy.

Background

The resistance to antibiotics developed by an increasing number of Is microorganisms is recognised to be a worldwide health problem (Tunger et al., 2000, Int. J. Microb. Agents 15:131-135; Jorgensen et al., 2000, Clin. Infect. Dis. 30:799-808). Thus, the development of non-antibiotic approaches for killing microorganisms is urgently required for controlling antibiotic-untreatable infections and limiting the development of additional antibiotic-resistant strains.

The treatment of microbial infections by photodynamic therapy (PDT) represents a valuable alternative method for eradicating bacteria since it involves a mechanism which is markedly different from that typical of >6 Mast Tibia Thus, PDT is based on the use of a photosensitizing molecule that, once activated by light, generates oxygen reactive species I'd?! it; #'{ :,,, j that are toxic for a large variety of prokaryotic and eukaryotic cells - 4ctild:: bacteria, mycoplasmas and yeasts (Malik et al., 1990, J. Photochem. Photobiol. B Biol. 5:281-293; Bertoloni et al., 1992, so Microbios 71:33-46). Importantly, the photosensitizing activity of many photodynamic agents against bacteria is not impaired by the resistance to antibiotics but, instead, depends mainly on their chemical structure (Maliketal.,1992,J. Photochem. Photobiol. BBiol. 14:262-266).

Various types of neutral and anionic photosensitising agents exhibit a pronounced phototoxic activity against Gram positive bacteria. However, such photosensitising agents exert no appreciable cytotoxic activity against Gram negative bacteria unless the permeability of the outer membrane is altered by treatment with ethylene diamine tetra-acetic acid lo (EDTA) or polycations (Bertoloni et al., 1990, FEMS Microbiol. Lett. 71: 149-156; Nitzan et al., 1992, Photochem. Photobiol. 55:89-97). It is believed that the cellular envelope of Gram negative bacteria, which is more complex and thicker than that of Gram positive bacteria, prevents an efficient binding of the photosensitising agent or intercepts and deactivates the cytotoxic reactive species photogenerated by the photosensitising agent (Ehrenberg et al., 1985, Photochem. Photobiol.

41:429-435; Valduga et al., 1993, J Photochem. Photobiol. B. Biol.

21:81-86).

so In contrast, positively charged (cationic) photosensitising agents, including porphyrins and phthalocyanines, promote efficient inactivation of Gram negative bacteria without the need for modifying the natural structure of the cellular envelope (Merchat et al., 1996, J. Photochem.

Photobiol. B. Biol. 32:153-157; Minnock et al., 1996, J. Photochem.

Photobiol. B. Biol. 32:159-164). It appears that the positive charge favours the binding of the photosensitising agent at critical cellular sites that, once damaged by exposure to light, cause the loss of cell viability (Merchat et al., 1996, J Photochem. Photobiol. B. Biol. 35:149-157).

Thus, it has been reported that Escherichia cold is efficiently inactivated so by visible light after incubation with the cationic 5,10, 15,20-tetrakis-(4 N-methylpyridyl)-porphine (T4MlFyP) (Valduga et al., 1999, Biochem. Biophys. Res. Commun. 256:84-88). The phototoxic activity of this porphyrin is mainly mediated by the impairment of the enzymic and transport functions of both the outer and cytoplasmic membranes, rather than by binding to DNA.

However, the utility of known porphyrin-based photodynamic therapy agents is limited due to their toxicity against mammalian host tissue cells, i.e. the compounds are unable to differentiate between target microbial lo cells and host cells. In addition, the utility of known porphyrin-based photodynamic therapy agents is further limited by their relatively low potency for target microbial cells. Furthermore, not all microbial infections are suitable for treatment using photodynamic therapy, for example the site of infection may not be accessible to light.

Hence, there is a need for new methods for killing microbial agents.

Summary

According to a first aspect of the invention, there is provided use of a compound of formula I in the preparation of a medicament for sonodynamic therapy X1 Y44415 z 1

X H AX

Y3S'Y2

I

wherein: X,, X2, X3 and X4 independently represent (i.e. are the same or different) a hydrogen atom, a lipophilic moiety, a phenyl group, a lower alkyl, alkaryl or aralkyl group, or a cationic group of the following formula; - L - R. - N (R2)(R3)R4 wherein: L is a linking moiety or is absent; Rat represents lower alkylene, lower alkenylene or lower alkynylene, which is optionally substituted by one or more substituents selected from lower alkyl, lower alkylene (optionally interrupted with oxygen), fluoro, ORs, C(0)R6, C(0)0R7, C(0) NR8 R9, NRIoRl and N+R2R3R4; and R2, R3 and R4 independently represent (i. e. are the same or different) H. aryl, lower alkyl, dower alkenyl or lower alkynyl, the latter three of which are optionally substituted by one or more substituents selected from lower alkyl, lower alkylene (optionally interrupted with oxygen), aryl, OR5, C(0)R6, C(0)0R7, C(0)NRg R9, NR'oR1 1 and o N-R'2Rl3Rl4 Z is -CH or N; Ye, Y2, Y3 and Y4 are absent or independently represent aryl, lower alkyl, lower alkenyl or lower alkynyl, the latter three of which are optionally substituted by one or more substituents selected from lower alkyl, lower alkylene (optionally interrupted with oxygen), aryl, OR5, C(0)R6, C(0)0R7, C(0) NR8R9, NRloRll and N+R,2R3R4; and R5, R6, R7, R8, R9, Rlo' R1', R12, R13 and R14 independently represent H or lower alkyl provided that at least one of X1, X2, X3 and X4 is a cationic group as defined above and at least one of X1, X2, X3 and X4 is a hydrogen atom, a phenyl group, a lipophilic moiety, or a lower alkyl, alkaryl or aralkyl group.

The term "lower alkyl" is intended to include linear or branched, cyclic or acyclic, C-C20 alkyl which may be interrupted by oxygen (preferably no more than five oxygen atoms are present in each alkyl chain). Lower alkyl groups which Rig, R2, R3, R4, R5, R6, R7, Rx, R9, Riot Rig, R'2, Rig and Rid may represent include C-Cg alkyl, C-C, alkyl, C-C4 alkyl, C-C2 alkyl, C-CO alkyl, C-Cg alkyl, C-C alkyl, C-C7 alkyl, C-C6 alkyl, C-C5 alkyl, CC4 alkyl, C-C3 alkyl and C-C2 alkyl. Preferred lower alkyl groups which Rat, R2, R3, R,+, R5, R6, R7, Rig, R9, R'o, Rat, Rig, Rig and Rid may represent include Car, C2, C3, C4, C5, C6, C7, Cx, C9, Call, lo Cal, Cal, Cal, Cot, C's and Cal alkyl.

Thus, any one or more of N+R2R3R4 and/or N+R'2R3R4 may represent cyclic amine/ammonium groups, for example: H R R R' R o C: :1:: :: O(j:1 I -I I -N+ -N+ -N+ + -I It will be appreciated that the cyclic amine/ammonium groups may also comprise fewer or greater than six members, for example such groups may comprise 4-, 5-, 7-, 8-, 9- or 1 O- membered rings.

The term "lower alkylene" is to be construed accordingly.

The terms "lower alkenyl" and "lower alkynyl" are intended to include linear or branched, cyclic or acyclic, C2-C20 alkenyl and alkynyl, respectively, each of which may be interrupted by oxygen (preferably no more than five oxygen atoms are present in each alkenyl or alkynyl chain).

The term "lower alkenyl" also includes both the cis and trars geometric isomers. Lower alkenyl groups which Ri, R2, R3, R4, R5,Ri, R7, Rs, R9, Ro, R, Rj2, R3 and Ri4 may represent include C2-Cx alkenyl, C2-C7 alkenyl, C2-C'6 alkenyl, C2-C4 alkenyl, C2-C2 alkenyl, C2-C,O alkenyl, C2-Cx alkenyl, C2-C7 alkenyl, C2-C6 alkenyl, C2-C5 alkenyl, C2-C4 alkenyl, C2C3 alkenyl and C3-C4 alkenyl. Preferred lower alkenyl groups which R, R2, R3, R4, R5, R6, R7, R8, R9, Rio, Ri, R'2, R3 and R4 may represent include C2, C3,C4,C5,C6, C7, C8, C9, C,, C, C,2, C3 and C4 alkenyl.

The term "lower alkenylene" is to be construed accordingly.

"Lower alkynyl" groups which R,R2, R3, R4,R5, R6, R7, Rs, Rg' Ro,R, R2, R3 and R,4 may represent include C2-C alkynyl, C2-C6 alkynyl, C2-C4 alkynyl, C2-C2 alkynyl, C2-C, alkynyl, C2-Cg alkynyl, C2-C alkynyl, C2-C7 alkynyl, C2-C6 alkynyl, C2-C5 alkynyl, C2-C4 alkynyl, C2 C3 alkynyl and C3-C4 alkynyl. Preferred lower alkynyl groups which R', R2, R3, R4, R5, R6, R7, R8, R9, Ro, R, R2, Ri3 and R4 may represent include C2, C3, C4'C5'C6'C7'C8'Cg,Cio'C,Cj2,Ci3 and C4 alkynyl.

The term "lower alkynylene" is to be construed accordingly.

The term "aryl" includes six to ten-membered carbocyclic aromatic groups, such as phenyl and naphthyl, which groups are optionally :5 substituted by one or more substituents selected from fluoro, cyano, nitro, lower alkyl (i.e. alkaryl), OR5, C(0)R6, C(0)0R7, C(0)NR8Rg and NRIoR 1 The term "aralkyl" includes aryl groups joined to the porphyrin ring via a o lower alkyl group.

A second aspect of the invention provides use of a compound of formula 1I in the preparation of a medicament for sonodynamic therapy Y4Z<1 X4 an/ / M X2 )=N N: Y3.JY2

II

wherein M is a metallic element or a metalloid element and A, X2, X3, X4, Y1, Y2, Y3, Y4 and Z are as defined above.

By "sonodynamic therapy", or SDT, we mean the medicament is capable lo of being activated upon exposure to ultrasound radiation. Thus, the active compound of the medicament is sonosensitive, such that it exhibits a cytotoxic effect upon ultrasound activation.

SDT is a comparatively new therapy, similar in concept to PDT, except that ultrasound radiation is used to activate a compound instead of light (see, for example, Misik et al., 1996, Free Rad. Res. 25:13-22 and Jeffers et al., 1995, J: Acoust. Soc. Am. 97:669-676). Although the mechanism underlying the cytotoxic action of sonosensitive compounds is not fully understood, it is believed to involve the formation of reactive radical so species by ultrasound, either as a result of direct pyrolysis in hot cavitation bubbles or after reaction with the OH radicals and H atoms which are generated by sonolysis of water.

In a preferred embodiment of the first and second aspects of the invention, the medicament is for killing or attenuating the growth of microorganisms. Preferably, the microorganisms are on a lightinaccessible surface or in a light-inaccessible area.

The term "metallic element" is intended to include a divalent or trivalent metallic element. Preferably, the metallic element is diamagnetic. More preferably, the metallic element is selected from Zn (II), Cu (II), La (III), Lu (III), Y (III), In (III) Cd (II), Mg (II), A1(III), Ru, Ni(II), Mn(III), lo Fe(III) and Pd(II). Most preferably, the metallic element is Ni(II), Mn(III), Fe(III) or Pd(II).

The term "metalloid" is intended to include an element having physical and chemical properties, such as the ability to conduct electricity, that are intermediate to those of both metals and non-metals. The term metalloid element includes silicon (Si) and germanium (Ge) atoms which are optionally substituted with one or more ligands.

It will be appreciated that the terms metallic element and metalloid element include a metal element or a metalloid element having a positive oxidation state, all of which may be substituted by one or more ligands selected from fluoro, OH, ORE wherein Rib is lower alkyl, lower alkenyl, lower alkynyl, aralkyl, aryl or alkaryl as defined above (wherein aryl and The compounds of formulae I and 11 comprise at least one cationic group.

Thus, the compounds may carry a net positive charge, for example a charge of +1, +2, +3, +4, +5, +6 or more. In a preferred embodiment, the compounds carry a net charge of less than +4, for example +1, +2 or +3.

In a particularly preferred embodiment, the compounds carry a net charge of+2.

It will be appreciated by persons skilled in the art that compounds of formulae I and II may be counterbalanced by counter-anions. Exemplary counter-anions include, but are not limited to, halides (e.g fluoride, chloride and bromide), sulfates (e.g. decylsulfate), nitrates, perchlorates, sulfonates (e.g. methane sulfonate) and trifluoroacetate. Other suitable counter-anions will be well known to persons skilled in the art. Thus, o pharmaceutically, and/or veterinarily, acceptable derivatives of the compounds of formulae I and It, such as salts and solvates, are also included within the scope of the invention. Salts which may be mentioned include: acid addition salts, for example, salts formed with inorganic acids such as hydrochloric, hydrobromic, sulfuric and phosphoric acid, with carboxylic acids or with organo-sulfonic acids; base addition salts; metal salts formed with bases, for example, the sodium and potassium salts.

It will be further appreciated by skilled persons that the compounds of so formula I may exhibit tautomerism. All tautomeric forms and mixtures thereof are included within the scope of the invention.

Compounds of formulae l and II may also contain one or more asymmetric carbon atoms and may therefore exhibit optical and/or diastereoisomerism. Diastereoisomers may be separated using conventional techniques, e.g. chromatography or fractional crystallization. The various stereoisomers may be isolated by separation of a racemic or other mixture of the compounds using conventional, e.g fractional crystallization or HPLC, techniques. Alternatively, the desired optical isomers may be made by reaction of the appropriate optically active starting materials under conditions which will not cause racemisation or epimerisation, or by derivatisation, for example with a homochiral acid followed by separation of the diastereomeric esters by conventional means (e.g. I-IPI,C, chromatography over silica). All stereoisomers are included within the scope of the invention.

In a preferred embodiment of the first and second aspects of the invention, Z is -CH.

lo A characterising feature of the compounds used in the first and second aspects of the invention is that at least one of substituent groups A, X2, X3 and X4 iS a quaternary ammonium cationic group of the formula -L RN(R2)(R3)R4, as defined above. Preferably, none of A, X2, X3 and X4 IS an anhnum or a pyrdnum caloric group.

In a preferred embodiment, Rut is an unsubstituted lower alkylene, lower alkenylene or lower alkynylene group.

Advantageously, Rut is a straight-chain lower alkylene group of formula: -(C112)m- Preferably, 'm' is an integer between 1 and 20. More preferably, 'm' is an integer between 1 and 10, for example between I and 6, between] and 5, between 1 and 4 or between 1 and 3. Preferred straight-chain lower alkylene groups which Rat may represent include groups of the above formula wherein m is 2, 3, 4, 5, 6, 7, 8, 9 or 10. Most preferably, 'm' is 2 or3.

The remaining three substituent groups of the quaternary ammonium moiety, i.e. R2, R3 and R4, may be the same or different and are selected from H. lower alkyl, lower alkenyl or lower alkynyl, the latter three of which are optionally substituted by one or more substituents selected from lower alkyl, OR5, C(0)R6, C(0)0R7, C(0)NRR9, NRoR and N FR2R3R4.

In a preferred embodiment, R2, R3 and/or R4 are lower alkyl, lower alkenyl or lower alkynyl group.

Preferably, R2, R3 and/or R4 are unsubstituted lower alkyl groups.

Optionally, at least one of R2, R3 and R4 is an alkyl group which is substituted with a primary, secondary or tertiary amine group or a quaternary ammonium group.

In a preferred embodiment of the first and second aspects of the invention, R is -(CH2)3-, R2 and R3 are CH3 and R4 is - (CH2)3 N(CH3)2 In an alternative preferred embodiment of the first and second aspects of the invention, R is -(CH2)3-, and R2, R3 and R4 are each CII3.

In a further alternative preferred embodiment of the first and second aspects of the invention, R. is -(CH2)3-, and R2, R3 and R4 are each C2H5 Advantageously, at least one of X, X2, X3 and X4 is a cationic group as defned above and at least one of X, X2, X3 and X4 is a hydrogen atom.

Preferably, each of X, X2, X3 and X4 iS a hydrogen atom or a cationic group as defined above.

Conveniently, the pK values of any primary, secondary or tertiary amine groups, if present in the compounds of the invention, is greater than 8 to ensure that the group is protonated when in a physiological environment.

The quaternary ammonium cationic group is optionally joined to the porphyrin ring via a linking moiety, L. lo Preferred linking moieties, L, include phenoxy, phenylene, sulfonyl amide, aminosulfonyl, sulfonylimino, phenylsulfonylamido, phenyl- aminosulfonyl, urea, urethane and carbamate linking moieties.

In a preferred embodiment, the quaternary ammonium cationic group is joined to the porphyrin ring via a phenoxy linker.

Thus, X, X2, X3 and/or X4 may have the following formula: '(OR)n wherein R is Rut - N+(R2)(R3)R4, as defined above, and 'n' is an integer between l and 3.

In an alternative preferred embodiment, the quaternary ammonium cationic group is joined to the porphyrin ring via a phenylene linker.

Thus, A, X2, X3 and/or X4 may have the following formula: Rm wherein R is Rat - N+(R2)(R3)R4, as defined above, and 'm' is an integer between 1 and 3.

Preferably, 'm' is 2, and most preferably 1.

In an alternative preferred embodiment, A, X2, X3 and/or X4 may have lo the following formula: (OR)n \/ \\ R m wherein R is Rat - N-(R2)(R3)R4, 'n' and 'm' are as defined above, and n + m' is between 1 and 3.

Advantageously, L comprises a benzene ring (e.g phenoxy, phenylene, phenylsulfonylamido or phenylamino-sulfonyl) mono-substituted at the para-position. Alternatively, L may be mono- or all-substituted at metaor or/ho-positions. L may also be bothpara- and or/ho-substituted.

so In an alternative preferred embodiment, the quaternary ammonium cationic group is joined directly to the porphyrin ring, i.e. L is absent.

In a preferred embodiment of the first and second aspects of the invention, the compound comprises two cationic groups, as defined :5 above, on opposite sides of the porphyrin ring, i.e. at ring positions 5 and or ring positions lO and 20. For example, Xl and X3 may be a hydrogen atom, a lipophilic moiety, a phenyl group, a lower alkyl, alkaryl or aralkyl group, and X2 and X4 may be cationic groups, or vice versa.

Preferably, X and X3 are both a hydrogen atom and X2 and X4 are both a cationic group, or vice versa.

Alternatively, the compound may comprise two cationic groups, as defined above, on neighbouring positions of the porphyrin ring, i.e. at ring positions 5 and IO, or ring positions 10 and 15, or ring positions 15 lo and 20 or ring positions 20 and 5. For example, X and X2 may be hydrogen and X3 and X4 may be cationic groups, or X2 and X3 may be hydrogen and X4 and X may be cationic groups, etc. It will be appreciated by persons skilled in the art that additional isomeric structural possibilities arise when Z represents nitrogen. Such possibilities are included within the scope of the present invention.

In a further preferred embodiment of the first and second aspects of the invention, the compound is substituted on one or more of its constituent so pyrrole rings. Thus, Ye, Y2, Y3 and Y4 may be absent or independently represent aryl, lower alkyl, lower alkenyl or lower alkynyl, the latter three of which are optionally substituted by one or more substituents selected from lower alkyl, lower alkylene (optionally interrupted with oxygen), aryl, OR5, C(0)R6, C(0)0R7, C(0)NRR', NRoR and N+R2R3R4. It will be appreciated by skilled persons that Ye, Y2, Y3 and/or Y4 may comprise cyclic groups, which may be saturated or aromatic. For example, one or more of the pyrrole rings may be substituted to form an iso-indole group, i.e. Ye, Y2, Y3 and/or Y4 together with the pyrrole ring to which they are attached may be cyclic.

In an alternative preferred embodiment of the first and second aspects of the invention, Ye, Y2, Y3 and Y4 are absent. Thus, the porphyrin ring is preferably substituted only at one or more of positions 5, 10, 15 or 20.

In a further preferred embodiment of the first and second aspects of the invention, at least one of X, X2, X3 and X4 iS or comprises a lipophilic moiety.

By 'lipophilic moiety' we include moieties having a partition coefficient lo between l-n-octanol and water expressed as log P of greater than 1.0 at physiological pH and 25 C.

Conveniently, the lipophilic moiety is a saturated, straight-chain alkyl group of formula - (CH2)pCH3, or an equivalent alkylene group of formula (CH2)p-, wherein 'p' is an integer between 1 and 22, for example between 1 and 18. Preferably, 'p' is between 1 and 18, more preferably between 2 and 16, between 4 and 16, between 6 and 18, between 8 and 16 or between 4 and 12. Most preferably, 'p' is between I O and 12.

It will be appreciated that X,, X2, X3 and/or X4 may be a cationic group, as defined above, which also comprises a lipophilic moiety.

In an alternative preferred embodiment of the first and second aspects of :5 the invention, none of X', X2, X3 and X4 iS a lipophilic moiety.

Advantageously, the compounds used in the first and second aspects of the invention are soluble in water. Preferably, the compounds may be dissolved in water to a concentration of at least 5 1lg/l, for example at least 10 g/1, 15,ug/1 or 20 g/1. More preferably, the compounds may be dissolved in water to a concentration of at least lOO 1lg/l, for example Ago, 300 g/1, 400 Owl, 500 g/1, 1 mg/ml, 5 mg/ml, lO mg/ml, mg/ml, SO ng/ml or lOO mg/ml.

Conveniently, the compounds used in the first and second aspects of the invention exhibit selective toxicity to microbial agents. By 'selective' we mean the compound is preferentially toxic to one or more microorganisms (such as bacteria, mycoplasmas, yeasts, fungi and/or viruses) compared to mammalian, e.g. human, host cells. Preferably, the lo toxicity of the compound to a target microorganism is at least two- fold greater than the toxicity of that compound to mammalian (e.g. human) cells, more preferably at least three-fold, at least four-fold, at least five- fold, at least six-fold, at least eight-fold, at least ten-fold, at least fifteen- fold or at least twenty fold. Most preferably, the compound of the invention is substantially non-toxic to mammalian cells.

In this way, when the compounds are used to treat bacterial infections, for example, dosing regimes can be selected such that bacterial cells are destroyed with minimal damage to healthy host tissue. Thus, the to compounds for use in the first and second aspects of the invention preferably exhibit a 'therapeutic window'.

Conveniently, the compounds of the invention exhibit greater toxicity to a target microorganism (e.g. a bacterium) upon ultrasound irradiation than as in the absence of activating irradiation. It will be appreciated that such toxicity may be determined using cultures of microbial cells. Preferably, the sonodynamic activity of a compound is at least two-fold greater than the innate toxicity of that compound, more preferably at least three-fold, at least four-fold, at least five-fold, at least sixfold, at least eight-fold, at so least ten-fold, at least i'ifteen-fold or at least twenty fold. Most preferably, the compound of the invention is substantially non-toxic in the absence of ultrasound irradiation.

In a preferred embodiment, the compound of the invention is toxic to the target microorganism (e.g. bacterial cells) at low doses. Preferably, the compound is toxic to the target microorganism at a concentration of less than 10 M, for example less than 1 M, less than 0.1 M, less than 0.01 M, less than 0.005 I1M or less than 0.001 M. lo Preferred compounds for use in the first and second aspects of the invention include the following: (a) 5,1 5-bis-(4- { 3-1 (3-Dimethylamino-propyl)-dimethyl-ammonio] propyloxy}-phenyl)-porphyrin dichloride ("Compound 8")

N_

1+ A/ \ 1+

-N- N N

Preferably, this compound is provided as a dichloride or tetrachloride salt.

(b) 5,1 5-bis-[4-(3-Triethylammonio-propyloxy)-phenyl]-porphyrin dichloride ("Compound 9") N>-O 0 Preferably, this compound is provided as a dichloride salt.

(c) 5,1 5-bis-[3-(3-Trimethylammonio-propyloxy)-phenyl]-porphyrin lo dichloride ("Compound 12") <NH Nit + )=/ IN HI \ + N./ - O it O N\Preferably, this compound is provided as a dichloride salt.

(d) 5,1 5-bis-[4-(3-Trimethylammonio-propyloxy)-phenyl]-porphyrin dichloride ("Compound 10")

O O

so Preferably, this compound is provided as a dichloride salt.

(e) 5-[3,5-bis-(3-Trimethylammonio-propyloxy)-phenyl]-1 5-undecyl porphyrin dichloride ("Compound 6") + N o 9 IN HN: + /_, 0 Preferably, this compound is provided as a dichloride salt.

(I) 5-{4-[3-Dimethyl-(3-dimethylaminopropyl)-ammonio 0 propyloxy]phenyl}1 5-(4-dodecyloxy-phenyl)-porphyrin chloride ("Compound 23")

-N

C12H2500 N._ Preferably, this compound is provided as a chloride or dichloride salt.

(g) 3-1 ({ 3-[(3- {4-[1 5-(4-Dodecyloxy-phenyl)-porphyrin-S-yl] phenoxy} propyl)-dimethyl-ammonio] -propyl} -dimethyl ammonio)-propyl]-trimethylammonium bichloride ("Compound 25") N -N.; C12H2500,Nx Preferably, this compound is provided as a bichloride salt. lo

(h) S. 1 5-bis-[3-(3-Trimethylammmonio-propyloxy)-phenyl]- l O undecylporphyrin dichloride ("Compound 28") 0> AN,N l5 Preferably, this compound is provided as a dichloride salt.

(i) 5- { 4- [3-Dimethyl-(3 -trimethylammonio-propyl)-ammonio- propyl oxy] -phenyl} -15 -(4-dodecyloxy-phenyl)-porphyrin dichloride ("Compound 31") IN 'art C12H25 RN '\\- O IN -N+ Preferably, this compound is provided as a dichloride salt.

(I) 5-[4-(3-Dimethyldecyl-ammoniopropyloxy)-phenyl]-15-{4-[3 0 dimethyl-(3-dimethylaminopropyl)-ammoniopropyloxy]-phenyl} porphyrin dichloride ("Compound 32") / -NH N O.--N. C H.

-N / Me2N

Preferably, this compound is provided as a dichloride salt.

It will be appreciated that the above compounds may alternatively be in a metallated form, i.e. they may comprise a chelated metallic element or metalloid element within the porphyrin ring.

The medicament as prepared according to the first or second aspects of the invention may be formulated at various concentrations, depending on the efficacy/toxicity of the compound being used and the indication for which it is being used. Preferably, the medicament comprises the compound at a concentration of between 0.1 EM and 1 rnM, more preferably between I EM and 100 M, between 5 EM and 50 I1M, between 10!1M and 50 juM, between 20,uM and 40 I1M and most preferably about 30 M. For in vitro applications, formulations may comprise a lower concentration of a compound, for example between lo 0.0025 EM and 1 M. It will be appreciated by persons skilled in the art that the compound used in the first or second aspects of the invention will generally be administered in admixture with a suitable pharmaceutical excipient diluent or carrier selected with regard to the intended route of administration and standard pharmaceutical practice (for example, see Remington: The Science and Practice of Pharmacy, 19 edition, 1995, Ed. Alfonso Gennaro, Mack Publishing Company, Pennsylvania, USA).

Suitable routes of administration are discussed below, and include topical, to pulmonary, nasal, aural, ocular, bladder and CNS delivery.

For example, for application topically, e.g to the skin or a wound site, the compounds can be administered in the form of a lotion, solution, cream, gel, ointment or dusting powder (for example, see Remington, supra, :5 pages 1586 to 1597). Thus, the compounds can be formulated as a suitable ointment containing the active compound suspended or dissolved in, for example, a mixture with one or more of the following: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax and water. Alternatively, so they can be formulated as a suitable lotion or cream, suspended or dissolved in, for example, a mixture of one or more of the following: mineral oil, sorbitan monostearate, a polyethylene glycol, liquidparaffin, polysorbate 60, cetyl esters wax, e-lauryl sulphate, an alcohol (e.g ethanol, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol) and water. s

In a preferred embodiment, the medicament (e.g lotion, solution, cream, gel or ointment) is water-based.

Formulations suitable tor topical administration in the mouth further include lo lozenges comprising the active ingredient in a flavoured basis, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert basis such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

Is The medicament as prepared according to the first or second aspects of the invention may also be administered intranasally or by inhalation and are conveniently delivered in the form of a dry powder inhaler or an aerosol spray presentation from a pressurised container, pump, spray or nebuliser with the use of a suitable propellant, JO e.g dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, a hydrofluoroalkane such as I, 1,1,2-tetrafluoroethane (HFA 134A3 or 1,1,1,2,3,3,3-heptafluoropropane (HFA 227EA3), carbon dioxide or other suitable gas. In the case of a pressurised aerosol, the dosage unit may be determined by providing a valve to deliver a metered :5 amount. The pressurised container, pump, spray or nebuliser may contain a solution or suspension of the active compound, e.g. using a mixture of ethanol and the propellant as the solvent, which may additionally contain a lubricant, e.g. sorbitan trioleate. Capsules and cartridges (made, for example, from gelatin) for use in an inhaler or insufflator may be formulated to contain a powder mix of a compound of the invention and a suitable powder base such as lactose or starch.

Aerosol or dry powder formulations are preferably arranged so that each metered dose or "puff'' contains at least 1 mg of a compound of the invention for delivery to the patient. It will be appreciated that the overall dose with an aerosol will vary from patient to patient and from indication to indication, and may be administered in a single dose or, more usually, in divided doses throughout the day. lo

Alternatively, other conventional administration routes known in the art may also be employed; for example the medicament as prepared according to the first or second aspects of the invention may be delivered orally, buccal]y or sublingually in the form of tablets, capsules, ovules, elixirs, solutions or suspensions, which may contain flavouring or colouring agents, for immediate-, delayed- or controlled-release applications. The medicament may also be administered intra-ocularly (see below), intra-aurally or via intracavernosal injection.

so The medicament may also be administered parenterally, for example, intravenously, intra-arterially, intraperitoneally, intrathecally, intraventricularly, intrasternally, intracranially, intra-muscularly or subcutaneously (including via an array of fine needles or using needlefree Powderject technology), or they may be administered by infusion :5 techniques. They are best used in the form of a sterile aqueous solution which may contain other substances, for example, enough salts or glucose to make the solution isotonic with blood. The aqueous solutions should be suitably buffered (preferably to a pH of from 3 to 9), if necessary. The preparation of suitable parenteral formulations under sterile conditions is readily accomplished by standard pharmaceutical techniques well known to those skilled in the art.

Formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents.

The formulations may be presented in unit-dose or multi-dose containers, lo for example sealed ampoules and vials, and may be stored in a freezedried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use.

Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.

The medicament may also be administered by the ocular route, particularly for treating diseases of the eye. For ophthalmic use, the compounds can be formulated as micronised suspensions in isotonic, pH adjusted, sterile saline, or, preferably, as solutions in isotonic, pH so adjusted, sterile saline, optionally in combination with a preservative such as a benzylalkonium chloride. Alternatively, they may be formulated in an ointment such as petrolatum.

For veterinary use, a compound is administered as a suitably acceptable :5 formulation in accordance with normal veterinary practice and the veterinary surgeon will determine the dosing regimen and route of administration which will be most appropriate for a particular animal.

The medicaments may be stored in any suitable container or vessel so known in the art. It will be appreciated by persons skilled in the art that the container or vessel should preferably be airtight and/or sterilised.

Advantageously, the container or vessel is made of a plastics material, such as polyethylene.

It will be appreciated by persons skilled in the art that the medicaments as prepared according to the first or second aspects of the invention may be used for killing and/or attenuating the growth of a number of types of microorganism, including bacteria, mycoplasmas, yeasts, fungi and/or viruses. It will be further appreciated that the medicaments may be used lo to prevent and/or treat infection with such microorganisms, i.e. the medicaments are suitable for prophylactic and/or therapeutic treatment.

For example, the medicament may be used to prevent or reduce the spread or transfer of a pathogen to other subjects, e.g. patients, healthcare Preferably, the medicaments as prepared according to the first or second aspects of the invention are for use in the curative and/or prophylactic treatment of bacterial infections such as Gram positive cocci (e.g. Streptococcus), Gram negative cocci (e.g. Neisseria), Gram positive so bacilli (e.g. Corynebacterium species), Gram negative bacilli (e.g. Escherichia colt), acid-fast bacilli (e.g. a typical Mycobacterium) and including infections causing abscesses, cysts, dermatological infections, wound infections, arthritis, urinary tract infections, pancreatitis, pelvic inflammatory disease, peritonitis, prostatitis, infections of the vagina, oral :5 cavity (including dental infections), eye and/or ear, ulcers and other localiscd infections; actinomyces infections; fungal infections such as Candida albicans, Aspergillus and Blastomyces; viral infections such as IIIV, encephalitis, gastro-enteritis, haemorrhagic fever, hantavirus, viral hepatitis, herpesvirus (e.g. cytomegalovirus, Epstein-Barr, herpesvirus so simiae, herpes simplex and varicella-zoster); protozoa! infections such as amoebiasis, babesiosis, coccidiosis, cryptosporidiosis, giardiasis, Leishmaniasis, Trichomoniasis, toxoplasmosis and malaria, helminthic infections such as caused by nematodes, cestodes and trematodes, e.g. ascariasis, hookworm, lymphatic filariasis, onchocerciasis, schistosomiasis and toxocariasis, and inflammatory diseases such as soft- tissue rheumatism, osteoarthritis, rheumatoid arthritis and spondyloarthropathies.

More preferably, the medicaments are for use in the curative and/or lo prophylactic treatment of infections by Gram positive bacteria and/or Gram negative bacteria. Most preferably, the compounds of the invention are for use in the curative and/or prophylactic treatment of infections by Gram positive bacteria.

The medicaments are preferably used to kill microorganisms, e.g. bacteria, mycoplasmas, yeasts, fungi and viruses. The medicaments are particularly suitable for killing bacteria which have developed resistance to conventional antibiotic treatments, such as methicillin-resistant Staphylococcus aureus (MRSA).

It will be appreciated by persons skilled in the art that the medicaments are suitable to treat microbial infections at any site capable of being irradiated with ultrasound, regardless of whether the site of infection is light accessible or not. Hence, such medicaments may have utility to treat infections which can not be treated by conventional photodynamic therapy agents. Preferably, the microbial infection is on a light- inacccssible surface or in a light-inaccessible area.

Dosages of the compound in the medicaments as prepared according to so the first or second aspects of the invention will depend on several factors, including the particular compound used, the formulation, route of administration and the indication for which the compound is used.

Typically, however, dosages will range from 0.01 to 20 mg of compound per kilogram of body weight, preferably from 0.1 to 15 mg/kg, for s example from 1 to 10 mg/kg of body weight.

In a preferred embodiment, the mcdicaments as prepared according to the first or second aspects of the invention are used in combination with conventional antimicrobial agents. For example, the compounds may be lo used in combination with one or more of the following conventional antibiotics: anti-bacterial agents, for example natural and synthetic penicillins and cephalosporins, sulphonamides, erythromycin, kanomycin, tetracycline, chloramphenicol, rifampicin and including gentamicin, ampicillin, benzypenicillin, benethamine penicillin, benzathine penicillin, phenethicillin, phenoxy-methyl penicillin, procaine penicillin, cloxacillin, flucloxacillin, methicillin sodium, amoxicillin, bacampicillin hydrochloride, ciclacillin, mezlocillin, pivampicillin, talampicillin hydrochloride, carfecillin sodium, piperacillin, ticarcillin, mecillinam, pirmecillinan, cefaclor, cefadroxil, cefotaxime, cefoxitin, cefsulodin o sodium, ceftazidime, ceftizoxime, cefuroxime, cephalexin, cephalothin, cephamandole, cephazolin, cephradine, latamoxef disodium, aztreonam, chlortetracycline hydrochloride, clomocycline sodium, demeclocydine hydrochloride, doxycycline, lymecycline, minocycline, oxytetracycline, amikacin, framycetin sulphate, neomycin sulphate, netilmicin, tobramycin, colistin, sodium fusidate, polymyxin B sulphate, spectinomycin, vancomycin, calcium sulphaloxate, sulfametopyrazine, sulphadiazine, sulphadimidine, sulphaguanidine, sulphaurea, capreomycin, metronidazole, tinidazole, cinoxacin, ciprofloxacin, nitrofurantoin, hexamine, streptomycin, carbenicillin, colistimethate, o polymyxin B. firazolidone, nalidixic acid, trimethoprim-sulfamethox mole, clindamycin, lincomycin, cycloserine, isoniazid, ethambutol, ethionamide, pyrazinamide and the like; anti-fungal agents, for example miconazole, ketoconazole, itraconazole, fluconazole, amphotericin, flucytosine, griseofulvin, natamycin, nystatin, and the like; and anti-vira] agents such as acyclovir, AZT, ddI, amantadine hydrochloride, inosine pranobex, vidarabine, and the like.

In a further preferred embodiment, the medicaments comprise and/or are coadministered with penetration enhancing agents, such as poly o (ethyleneimine), or antibiotic agents which exhibit such penetrationenhancing capability (e.g. polymyxin or colistin).

The medicaments as prepared according to the first or second aspects of the invention are particularly suited for use in the curative or prophylactic treatment of one or more of the following indications: Impetigo Impetigo is a highly communicable infection. It is the most common infection in children.

Impetigo have two classic forms nonbullous and bullous. The nonbullous impetigo, also named impetigo contagiosa accounts for approximately 70% of cases. Lesions normally resolve in 2 to 3 weeks without treatment. Impetigo also may complicate other skin diseases such as scabies, varicella, atopic dermatitis, and Darier's disease.

(a) Nonbullous Impetigo Type of bacteria s Nonbullous is an infection caused principally by Group A beta- haemolytic streptococci (Streptococcus pyogenes), Staphylococcus aureus, or a combination of these two organisms (see Andrews' diseases of the skin: clinical dennatology 9th ed. (2000) edited by Odom RB editor Saunders p.3 12-4). Non-Group A (Group B. C, and G) lo streptococci may be responsible for rare cases of impetigo, and Group B streptococci are associated with impetigo in the newborn.

Type of wounds Nonbullous impetigo is a superficial, intraepidermal, unilocular vesiculopustular infection.

Lesions of nonbullous impetigo commonly begin on the skin of the face or extremities following trauma. As a rule, intact skin is resistant to so impetiginazation.

The clinical presentation of impetigo evolves in an orderly fashion from a small vesicle or pustule, which progresses into honey-coloured crusted plaque. Lesions usually are less than 2 cm in diameter. I,esions tend to dry, leaving fine crusts without cicatrisation. Lesions are usually minimally symptomatic. Rarely, erythema associated with mild pain or slight pruritus may be present. The infection spreads to contiguous and distal areas through the inoculation of other wound from scratching.

Site of bacteria Nonbullous impetigo is a superficial streptococcal or staphylococcal infection which is localised to the subcorneal (just beneath the stratum corneum) layer of the skin (see Figure 1). More particularly, infection in impetigo is confined histopathogically to highly differentiated, upper epidermal keratinocytes. Once the bacteria invade a break in the skin, they begin to multiply.

lo The histopathology is that of an extremely superficial inflammation about the funnel-shaped upper portion of the pilosebaceous follicles. A subcorneal vesicopustule is formed, containing a few scattered cocci, together with debris of polymorphonuclear leukocytes and epidermal cells. In the dermis, there is a mild inflammatory reaction - vascular dilatation, oedema, and infiltration of polymorphonuclear leukocytes (Andrews' diseases of the skin, supra., p.3 12-4).

(b) Bullous impetigo Type of bacteria Bullous impetigo is caused primarily by strains of Staphylococcus aureus which produce exfoliative toxins (Sadick et al., 1997, Dermatologic Clinics 15(2): 341-9).

Type of wounds Bullous impetigo is histologically characterised by subcorneal cleavage and infiltrate with polymorphonuclear leucocytes migrating through the so epidermis and accumulating between granular and stratum corneum skin layers. Small or large superficial fragile bullae are present on the trunk and extremities.

Flaccid bullae and moist erosions with surrounding erythema are s characteristic of this subcorneal infections. Often, only the remnants of ruptured bullae are seen at the time of presentation. The separation of the epidermis is due to an exotoxin produced by Staphylococcus aureus.

Sites of bacteria Bullous impetigo is a superficial staphylococcal infection that occurs in and just beneath the stratum corneum (see figure 1). Bullous impetigo is considered due to exfoliative toxin produced by some Staphylococcus aureus attached to stratum corneum cells.

Atopic dermatitis (AD! Atopic dermatitis, also named atopic eczema, is a chronic inflammation of the skin resulting in an itchy rash, especially in the flexures i.e. behind to the knees, in front of the elbows, wrists, neck, and eyelids. Infection of the rash is common, and causes further inflammation and itch.

Eczema typically manifests in those aged 1-6 months. Approximately 60% of patients have their first outbreak by 1 year and 90% by 5 years.

:5 Onset of atopic dermatitis in adolescence or later is uncommon and should prompt consideration of another diagnosis. Disease manifestations vary with age.

Type of bacteria Bacteria and their superantigens contribute to the pathogenesis of AD.

s Staphylococcus aureus colonises the skin of 90% of AD patients (chronic eczematous lesions) and only 5% of non-atopic patients. The colonisation density of Staphylococcus aureus can reach up to 107 colony forming units cm2 without clinical signs of infection in patients with AD.

In addition, the apparently normal non-lesional skin of atopic patients lo contains increased numbers of Staphylococcus aureus.

The reason for the overgrowth of Staphylococcus aureus in atopic dermatitis, though much less severely or not at all in diseases such as psoriasis, is not known. Protein A elicits a much less vigorous response in atopics than in normals or psoriatics, but this may be the result rather than a cause of colonization. Attention has recently turned to the skin lipids and there is some evidence that fatty acids which may control staphylococcal colonization are deficient in atopics.

so Superantigens are a unique group of proteins produced by bacteria and viruses that bypass certain elements of the conventional, antigenmediated immune sequence. Whereas conventional antigens activate approximately 0.01% to 0. l % of the body's T cells, a superantigen has the ability to stimulate 5% to 30% of the T-cell population. S. aureus may as exacerbate or maintain skin inflammation in AI) by secreting a group of exotoxins that act as superantigens. AD patients possess an altered skin barrier secondary to an insufficiency of ceramides within the stratum corneum. It has been proposed that penetration of the skin by these exotoxins may cause activation of T cells, macrophages, LCs, and mast so cells, thereby leading to the release of cytokines and mast cell mediators.

It is conceivable that these events may provide the basis for inflammation in chronic AD. Speculation remains whether S. aureus colonisation and local superantigen secretion is a primary or secondary phenomenon in AD (Andrews' diseases of skin, Chap. 5, Atopic Dermatitis, Eczema, and non-infectious immunodeficiency disorders, p.69-76).

Cutaneous viral, fungal, and bacterial infections occur more commonly in AD patients. Viral infections are consistent with a T cell defect and include herpes simplex (local or generalised, i.e. eczema herpeticum), lo molluscum contagiosum, and human papilloma virus. Superficial fungal infections with Trichophyton rubrum and Pityrosporon ovate also occur frequently. Bacterial infections, specifically those with S. aureus, are extremely common. Superinfection results in honey-coloured crusting, extensive serous weeping or folliculitis.

Type of wounds Acute lesions appear as erythematous papules, vesicles, and erosions; chronic disease consists of fibrotic papules and thickened, lichenified skin.

A finding of increasing numbers of pathogenic staphylococci is frequently associated with weeping, crusting, folliculitis and adenopathy.

Secondary staphylococcal infection is frequent and local oedema and :5 regional adenopathy commonly occur during atopic dermatitis. Impetigo can be a sort of secondary infection of atopic dermatitis.

The histology of atopic dermatitis ranges from acute spongiotic dermatitis to lichen simplex chronicus, depending on the morphology of the skin so lesion biopsied.

Sites of bacteria Staphylococcus aureus cell walls exhibit receptors, the so-called s adhesins, for epidermal and dermal fibronectin and fibrinogen. It has been demonstrated that the binding of Staphylococcus aureus was mediated by fibrinogen and fibronectin in AD patients. As the skin of AD patients lacks an intact stratum corneum, dermal fibronectin might be uncovered and increase the adherence of Staphylococcus aureus. Fibrillar lo and amorphous structures have been traced between Staphylococcus aureus cells and corneocytes and may results in a bacterial biofilm. It has been observed that Staphylococcus aureus penetrates into intracellular spaces suggesting that the skin surface lipids are deteriorated in AD patients (see Breuer K e' al., 2002, British Journal of Dermatology 147: 55-61).

Ulcers Skin ulcers, such as diabetic foot ulcers, pressure ulcers, and chronic venous ulcers, are open sores or lesions of the skin characterized by the wasting away of tissue and sometimes accompanied by formation of pus.

Skin ulcers may have different causes, and affect different populations, but they all tend to heal very slowly, if at all, and can be quite difficult and expensive to treat.

Type of bacteria Superficial pressure ulcers are not associated with major infection problems. Aerobic microorganisms at low levels will contaminate so pressure ulcers, but will not impede timely healing. However, deep full thickness pressure ulcers can become secondarily infected, and osteomyelitis can occur. Those pressure ulcers with necrotic tissue contain high levels of aerobic and anaerobic microorganisms as compared to non-necrotic ulcers, foul smell is usually present when anaerobes invade the tissues. Thus, a treatment strategy is to clear necrotic tissue from the wound, producing a decrease in anaerobe presence.

The infections of pressure ulcers are typically polymicrobial and can contain Streptococcus pyogenes, enterococci, anaerobic streptococci, lo Enterobacteriaece, Pseudomonas aeruginosa, Bacteroides fragilis and Staphylococcus aureus.

Type of wounds Stage I pressure ulcer: Nonblanchable erythema of intact skin, considered to be heralding lesion of skin ulceration.

Stage II pressure ulcer: Partial thickness skin loss involving the epidermis and/or dermis. The ulcer is superficial and presents clinically as an JO abrasion, blister, or shallow crater. Because the epidermis may be interrupted by an abrasion, blister, or shallow crater, the ulcer should be evaluated for signs of secondary infections.

Stage ITI: Full thickness skin loss involving damage or necrosis of :5 subcutaneous tissue which may extend down to, but not through, underlying fascia. The ulcer presents clinically as a deep crater with or without undermining of adjacent tissue.

Stage IV: Full thickness skin loss with extensive destruction, tissue necrosis, or damage to muscle, bone, or supporting structures, such as tendons or joint capsules.

Sites of bacteria There are three microbiological states that are possible in a wound: contamination, colonization and infection. Contamination is characterized as the simple presence of microorganisms in the wound but without lo proliferation. It is generally accepted that all wounds, regardless of aetiology, are contaminated. Colonisation is characterized as the presence and proliferation of microorganisms in the wound but without host reaction. Colonisation is a common condition in chronic wounds such as venous ulcers and pressure ulcers and does not necessarily delay the healing process. When bacteria invade healthy tissues and continue to proliferate to the extent that their presence and by-products elicit or overwhelm the host immune response, this microbial state is known as infection. The classic signs and symptoms of infection include local redness, pain and swelling, fever and changes in the amount and character of wound exudates.

Lung infections The compounds of the invention are also suitable for treating a patient :5 having an infectious disease of the lung, by administering to the subject a compound of the invention and irradiating the lung with ultrasound having a intensity that causes the compound to produce an anti- microbial effect. Lung infection can occur with a variety of bacterial genera and species, which include Mycobacterium tuberculosis (tuberculosis), so Pseudomonas (primary cause of death of cystic fibrosis patients), Streptococcus, Staphylococcus pneumonias, Klebsiella, Toxoplasma, etc. Lung infection can also occur with a variety of virus strains and opportunistic pathogens (fungi, parasites). As pathogens of the lung are increasingly resistant to classical antibiotic therapies, sonodynamic therapy offers an alternative method for eliminating these harmful organisms.

The compound of the invention can be administered to the lung in a variety of ways. For example the compound can be administered by the lo respiratory tract (i.e. intra-tracheally, intra-bronchially, or intraalveolarly) or through the body wall of the chest. The ultrasound source can be applied externally or through these routes as well with the help of flexible probes for example. The irradiation can be directed to the base of the lung, to the apex of the lung, or both.

Further indications The compounds of the invention are also suitable for the curative and/or prophylactic treatment of the following: Infections of burn sites and skin grafts; otitis (ear infection), bacterial conjunctivitis and other eye infections, periodontitis and other dental infections, and infected bones exposed during surgical procedures.

Cancerous cells and tissue (including turnouts of the head, neck, gastrointestinal tract, liver, breast, prostate, lung and malignancies of haenatopoietic and lymphoid cells), other neoplastic disorders (such as atherosclerosis and restenosis), acne, psoriasis and actinic keratosis.

Thus, further aspects of the invention provide the following: (i) Use of a compound as described above in the preparation of a medicament for the curative and/or prophylactic treatment of a dermatological infection; (ii) Use of a compound as described above in the preparation of a medicament for the curative and/or prophylactic treatment of an infection of the lungs; (iii) Use of a compound as described above in the preparation of a medicament for the curative and/or prophylactic treatment of a (iv) Use of a compound as described above in the preparation of a medicament for the curative and/or prophylactic treatment of cancerous cells and/or tissues; and (v) A method for treating a patient in need of treatment with a no sonodynamic therapy agent comprising administering to the patient a compound as defined above in relation to the first and/or second aspects of the invention and irradiating the compound with ultrasound; and :5 (vi) A method for preventing wound infection comprising contacting the wound with a compound of the invention and irradiating the compound with ultrasound.

To treat patients in accordance with the methods of the invention, the so medicament is exposed to an amount of ultrasound acoustic energy sufficient to generate a cytotoxic effect. Preferably, the frequency and power levels are sufficient to produce ultrasonic cavitation or mechanical shearing in the body.

Generally, ultrasound will be delivered using one or more of the following parameters: (a) a power level (intensity) of about 0.1 W/cm2 to about 10000 W/cm2, preferably about 0.1 to 500 W/cm2, 0.5 to lOO W/cm2, 0.1 to lo 20 W/cm2 or about 0.25 to 40 W/cm2 (more preferably, an intensity of about 4 to 12 W/cm2 or about 2 to 8 W/cm2 is used); (b) a frequency of about 1 kHz to about 500 MHz, for example about 0.01 to about 100 MHz, preferably about 0.1 to about 40 MHz or about 0.05 to 12 MHz (more preferably, a frequency of about 0.01 to MHz or 0.1 to 10 MHz is used, for example about 0.5 to 5 MHz or about l to 3 MHz); and (c) a duration of lOmsec to 5 hours, preferably about l second to JO 60 minutes, for example about l minute to 40 minuses or about l minute to 5 minutes.

Persons skilled in the art will readily appreciate that these values depend on the transducer frequency, type of tissue being irradiated and sonodynamic compound use. The ultrasound can be pulsed, second harmonic or continuous wave, and can be delivered using commercially available diagnostic or therapeutic devices.

Preferably, the dose of ultrasound radiation is sufficient to increase or so enhance the antimicrobial activity of the active compound in the medicament compared to its activity in the absence of exposure to ultrasound radiation.

It will be appreciated by persons skilled in the art that ultrasound irradiation may take place at various time points after application of the compound of the invention. Typically, the compound is irradiated between 5 minutes and 24 hours after application, for example, between 5 minutes and2 hours or between 10 minutes and 1 hour. Optimal irradiation times may be determined by experimentation.

The medicaments prepared according to the first and second aspects of the invention may also be used to kill microorganisms in vitro. Thus, the medicaments may also be used in the form of a sterilising solution or wash to prevent the growth of microorganisms on a surface, for example in a clinical environment (e.g. surgical theatre) or a domestic environment (e.g. a kitchen work surface).

Preferably, such a medicament comprises the antimicrobial compound in solution at a concentration of 1 to 100 g/ml.

Preferably, the solution further comprises a surface-active agent or surfactant. Suitable surfactants include anionic surfactants (e.g. an aliphatic sulphonate), amphoteric and/or zwitterionic surfactants (e.g. derivatives of aliphatic quaternary ammonium, phosphonium and sulfonium compounds) and nonionic surfactants (e.g. aliphatic alcohols, acids, amides or alkyl phenols with alkylene oxides) Conveniently, the surface-active agent is present at a concentration of 0.5 to 5 weight percent.

The sterilising solutions are particularly suited for use in hospital environments. For example, the sterilising solutions may be used to sterilise surgical instruments and surgical theatre surfaces, as well as the hands and gloves of theatre personnel. In addition, the sterilising solutions may be used during surgery, for example to sterilise exposed bones. In all cases, the solution is applied to the surface to be sterilised and then /irradiated so as to produce a reactive oxygen species (see above) lo In both in vitro and in viva uses, the medicament prepared according to the first and second aspects of the invention is preferably exposed to the target microorganisms (or surface/area to be treated) for at least five minutes. For example, the exposure time may be at least 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, l hour, 2 hours, 3, hours, 5 is hours, 12 hours and 24 hours.

Preferred, non-limiting embodiments of the invention will now be described by way of example, with reference to the accompanying JO drawings in which: Figure 1 shows a schematic diagram of the structure of skin.

Figure 2 shows the chemical stability of Compound 10 formulated (A) as :5 a solid, (B) in water and (C) in PBS.

Figure 3 shows a 3D plot of the stability (measured by HPLC) of Compound 10 after 21 days in PBS buffer.

Figure 4 shows the stability over 8 weeks of various formulations of (A) Compound 1, (B) Compound 8, (C) Compound 12 and (D) Compound 10.

Figure 5 shows the extended stability over 17 weeks of various formulations of (A) Compound 10 and (B) Compound 8.

EXAMPLES

SYNTHESIS OF EXEMPLARY COMPOUNDS

Materials and Methods NMR-measurements Proton NMR spectra were recorded on a Bruker B-ACS60 (300 MHz) o instrument using TMS as internal standard. The chemical shifts are given in ppm and coupling constants in Hz in the indicated solvent. Some abbreviation for NMR: singlet (s), broad singlet (bs), doublet (d), triplet (t), quartet (q), quintet (quint), multiplet (m).

Chemicals All solvents and reagents were purchased from Aldrich, Fluka, Merck and Lancaster and used without further purification.

so Dipyrrolmethane was prepared as described by C. Brucker et al., J; Porphyrins Phthalocyanines, 2 455 (1998).

Chromatography :5 Column chromatography was carried out using silica gel (Merck Silicagel 60, Fluka 60, 0.040-0.063 mm) and Sephadex LH-20 (Pharmacia). All solvents (Synopharm) for chromatography were technical pure grade.

Abreviations DDQ: 2,3-dichloro-5,6-dicyano-p-benzoquinone DMF: N,Ndimethylformamide TEA: trifluoroacetic acid Synthesis routes for test compounds The following test compounds were synthesised: Exemplary compounds for use in the invention Compounds 6,8 to 1O, 12, 23, 25, 28, 31 and 32.

Reference compounds (for use as comparative controls) Compounds 1,3, 16, 19, 26, 29, 33, 36, 37, 39, 41 and 46 to 51.

Chemical intermediates Compounds 2, 4, 5, 7, 11, 13 to 15, 17, 18, 20 to 22, 24, 27, 30, 34, 35, 38, 40 and 42 to 45.

COMPOUND 1 5,1 On 1 5,20-tetrakis-[4-(3-Trimethylammonio-propyloxy)-phenyl]- porphyrin tetrachloride O--,N

_ N >\ 0!

- N O. N

To a vigorously-stirred suspension of 5,1 0,1 5,20-tetrakis-(4-hydroxy phenyl)-porphyrin (50 ma, 0.07 mmol) and K2CO3 (230 ma, 1.7 mmol) in DMF (20 mL), a solution of ( 1 -bromopropyl)-trimethylammonium lo bromide (0. 27 g, 1.05 mmol) in DMF (5 mL) is added dropwise at 50 C during 30 mins. The mixture is stirred at 50 C for 15 h. After removal of DMF under reduced pressure, the residue obtained is dissolved in methanol (5 mL) and filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After washing with methanol (1 L), the pad is eluted with acetic acid. After evaporation of solvent from the eluate, the residue obtained is purified by chromatography on a column (2.5 x 40 cm) of Sephadex LH20 elating with n butanol:water:acetic acid (4:5:1, by vol., upper phase). The recovered material is dissolved in the minimum volume of methanol and the so solution is passed through a short column (3.5 x 20 cm) of anion exchange resin (Amberlite IRA 400, chloride form). The recovered tetrachloride salt is dried under high vacuum and obtained as a violet solid.

H-NMR: OH (300z, CD3OD): 2.35-2.50 (bs, 8 H), 3.25-3.35 (bs, 36 H), 3.65 3.75 (bs, 8 H), 4.35 (m, 8 H), 7.30, 8.10 (2 x d, 318.5 Hz, 16 H), 8.80 9.00 (bs, 8 H).

COMPOUND 2 5,10, ] 5-tris-(4-Hydroxy-phenyl)-20-(4-undecyloxy-phenyl)-porphyrin

OH HO,

OH

To a vigorously-stirred suspension of 5,10,15,20-tetrakis-(4-hydroxyphenyl)-porphyrin (400 ma, 0.59 mmol) and K2CO3 (1.0 g, 7.1 mmol) in DMF (75 mL), a solution of l-bromoundecane (0.1 mL, 0.45 mmol) in DMF (10 mL) is added dropwise at 50 C during 30 mins and the mixture is stirred at the same temperature for 1.5 h. After removal by filtration of K2CO3 and removal under reduced pressure of DMF, the residue obtained is dissolved in dichloromethane (200 mL), washed with water (3x150 mL) and the solution dried (Na2SO4). The solvent is evaporated under reduced pressure and the residue obtained is dissolved in toluene:ethanol (5:1 by vol., cat 10 mL) and purified by chromatography using a column (5 X 50 cm) of silica gel (Merck 60). The column is eluted with toluene followed by toluene:ethyl acetate (2:1 by vol.) and the desired material recovered by evaporation of solvent from the appropriate fractions is dried under high vacuum. The product is obtained as a violet solid.

IH-NMR: OH (300Mz, d6-acetone): 0.95 (t, 3J 7.5 Hz, 3 H), 1.25-1.55 (m, 14 H), 1.58 (quint, 3J7.5 Hz, 2 H), 1.85 (quint, 3J 7.5 Hz, 2 H), 4.16 (t, 3J 7. 5 Hz, 2 H), 7.20 (d,3J8.1 Hz, 2 11), 7.25 (d,3J8.2 Hz, 6 H), 8.00-8.15 (m, 8H), 8.80-9.10 (m, 8H).

COMPOUND 3 5,10,15-tris-[4-(3-Trimethylammonio-propyloxy)-phenyl]-20-(4undecyloxy-phenyl)-porphyrin bichloride t

N >\

0}}0'' +

O N

To a vigorously-stirred suspension of Compound 2 (100 ma, 0.12 mmol) and K2CO3 (230 ma, 1.7 mmol) in DMF (30 mL), a solution of (1bromopropyl)-trimethylammonium bromide (0.3 g, 16.6 mmol) in DMF (10 mL) is added at 50 C and the mixture is stirred at this temperature for 12 h. After removal of the DMF under reduced pressure, the residue obtained is dissolved in methanol (5 mL) and filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After o washing with methanol (ca. 1L), the pad is eluted with acetic acid:methanol:water (3:2:1, by vol.). After evaporation of the solvent from the eluate under reduced pressure, the residue obtained is purified by chromatography on a column (2.5 x 40 cm) of Sephadex LH-20 eluting with n-butanol:water:acetic acid (5:4:1, by vol., upper phase). After removal of the solvent from appropriate fractions of the eluate under reduced pressure, the residue obtained is dissolved in methanol (5 mL) and the solution is passed through a short column (3.5 x 20 cm) of anion exchange resin (Amberlite IRA 400, chloride form). The final product is obtained as the bichloride salt, after removal of solvent and drying under high vacuum, as a violet solid.

IH-NMR: OH (300MHz, CD3OD): 0.80 (I, 3J 7.5 Hz, 3 H), 1.15-1.45 (m, 16 H), 1.50-1.60 (bs, 2 H), 2.25-2.45 (bs, 6 H), 3.25-3.35 (bs, 27 H), 3.75-3.85 (bs,, 6 H), 4.18 (t, 3J 7.5 Hz, 2 H), 4.40-4.45 (bs, 6 H), 7.20-7.40, 7. 95 8.15 (2 x m, 16 H), 8.60-9.00 (bs, 8 H).

COMPOUND 4 5-(3,5 -Dimethoxy-phenyl)- 15-undecyl-porphyrin MeO To a stirred solution of dipyrrolemethane (0.62 g, 4.2 mmol) in dichloromethane (5 mL) is added 3,5-dimethoxybenzaldehyde (0.35 g, 2.1 mmol) and dodecanal (0.464 g, 2.52 mmol) in degassed dichloromethane (IL). TEA (0.07 mL, 3.0 mmol) is added dropwise.

The solution is stirred at room temperature in the dark for 17 h under argon. After addition of DDQ (2.7 g, 12 mmol), the mixture is stirred at room temperature for a further hour. Purification of material recovered after removal of solvent under reduced pressure by chromatography on a column (400 g) of silica gel (Merck 60) with toluene for elusion yields the product as a violet solid.

]H-NMR: 611 (300Mz, CDCI3): 0.80 (t, 3J7.5 Hz, 3 H), 1.]0-1.25 (m, 12 H), 1.40 (m, 2H), 1.75 (quint,, 3J7.5 Hz, 2 II), 2.45 (quint,3J7.5 Liz, 2 H), 3.90 (s, 6H), 4.90 (t,3J 7.5 Hz, 2 H), 6.80 (m, 1 H), 7.35 (m, 2 H), 9.00, 9. 25, 9.30,9.50(4xd,,3J4.7 Hz, 4x2H), ]0.15(s,2H).

COMPOUND 5 5-(15-Undecyl-porphyrin-5-yl)-benzene- 1,3-diol HO 9 NH Nor A

HO

To a solution of Compound 4 (80 ma, 0.133 mmol) in anhydrous dichloromethane (80 mL) under an argon atmosphere, BBr3 (5 mL, 1M in dichloromethane) is added dropwise at-70 C and the mixture is stirred for 1 h at this temperature and then warmed to room temperature and stirred overnight. The mixture is cooled to -10 C and hydrolysed by the addition of water (2 mL) and stirring for 1 h. NaHCO3 (3 g) is added directly for neutralization. The mixture is stirred for a further 12 h and after filtration of NaHCO3 and removal of dichoromethane under vacuum the residue obtained is purified by column chromatography using silica gel eluting with dichloromethane. After evaporation of solvent from appropriate combined fractions and drying of the residue obtained under high vacuum the product is obtained as a violet solid 2s 'H-NMR: (300Mz, d6-acetone): 0.75 (t, 3J 7.5 Hz, 3 H), 1.05-1.25 (m, 12 H), 1.30-1.40 (m, 2H), 1.45-1.50 (m, 2 I-l), 2.40 (quint, 3J 7.5 Hz, 2 H), 4. 90 (t, 3J 7.5 Hz, 2 H), 6.65 (m, 1 H), 7.18 (m, 2 H), 8.60-8.65, 9.00-9.05, 9.35-9.40, 9.55-9.60 (4 x m, 8 H), 10.25 (s, 2H).

COMPOUND 6 5-[3,5-bis-(3-Trimethylammonio-propyloxy)-phenyl]-15-undecyl porphyrin dichloride + IN-o W9 C 1 1 H23 + 0 lo To a vigorously-stirred suspension of Compound 5 (80 ma, 0.14 mmol) and K2CO3 (230 ma, 1.7 mmol) in DMF (30 mL) is added (1 bromopropyl)-trimethylammonium bromide (0.3 g, 16.6 mmol) at 50 C.

The mixture is stirred at this temperature for 18 h. After removal of the DMF under reduced pressure, the residue obtained is dissolved in methanol (5 mL) and filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After washing the pad with methanol (ca. I L) the crude product is eluted with acetic acid:methanol:water (3:2:1, by vol.). Appropriate fractions are collected and, after evaporation of the solvent under reduced pressure, the residue go obtained is purified by chromatography on a column (2.5 x 40 cm) of Sephadex LH-20 eluting with n-butanol:water:acetic acid (5:4:1, by vol., upper phase). After removal of the solvent from appropriate fractions under reduced pressure, the residue obtained is dissolved in methanol (5 mL) and the solution is passed through a short column (3.5 x 20 cm) of :5 anion exchange resin (Amberlite IRA 400, chloride form). After collection of the eluate, solvent is removed under reduced pressure and the residue obtained is dried under high vacuum to yield the dichloride salt as a violet solid.

IH-NMR: 3,, (300Mz, CD3OD): 0.75 (t,317.5 Hz, 3 H), 1.05-1.20 (m, 14 H), 1.45 1.50 (m, 2 H), 2.05-2.15 (m, 4 H), 2.15-2.20 (m, 2 H), 2.95 (s, 18 H), 3.35-3.45 (m, 4 H), 3.95 (t, 317.5 Hz, 4 H), 4.55 (t,3J 7.5 Hz, 2 H), 6. 85 (m, 1 T]), 7.35 (m, 2 H), 8.85-8.90, 9.15-9.20, (3 x m, 8 H), 10.10 (s, 2 H).

lo COMPOUND 7 5,15-bis-[4-(3-Bromo-propyloxy)-phenyl]-porphyrin Br B: To a stirred solution of dipyrrolemethane (0.61 g, 4.1 mrnol) and 4-(3 bromopropyloxy)- benzaldehyde (1.03 g, 4.2 mmol) in degassed dichloromethane (I L), TFA (0. 07 mL, 1.5 mmol) is added dropwise.

The solution is stirred at room temperature in the dark under argon for 17 h. After addition of DDQ (2.76 g, 0.012 mol), the mixture is stirred at o room temperature for a further hour. Filtration through silica gel (Fluke 60, 100 g) using dichloromethane for elusion gives raw product which, after treatment with dichloromethane:n-hexane, yields pure product as a violet solid.

1H-NMR: 811 (300Mz, C6D6): -3.] 5 (2 H. s), 2.00 (quiet, 3J 7.5 Hz, 4 H), 3.30 (t, 3J 7.5 Hz, 4 H), 3.90 (t, 3.17.5 Hz, 4 H), 7.15-7.18, 7.95-8.15 (2 x m, 2 x 4 H), 9.15-9.20,(m, 8 H), 10.05 (s, 2H).

COMPOUNI) 8 5,15 -bis-(4- { 3 -[(3 -Dimethylamino-propyl)-dimethyl-ammonio] propyloxy} -phenyl)-porphyrin dichloride

N_

1+ - \ 1+

-N- N N

(0{}0) lo Compound 7 (200 ma, 0.27 mmol) is dissolved in absolute DMF (40 mL) with N,N,N',N'-tetramethyl-1,3-propanediamine (5 mL, 13,9 mmol) and the solution is stirred at 50 C under argon overnight. After evaporation Is of the solvent under reduced pressure, the residue obtained is dissolved in methanol (5 mL) and the solution is filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). The pad is eluted with methanol (ca. 1L) followed by acetic acid:methanol:water (3:2:1, by vol.). After evaporation of the solvent from appropriate o fractions, the raw product obtained is dissolved in methanol (5 mL) and further purified by chromatography on a column (2.5 x 40 cm) of Sephadex LH-20 using n-butanol:water:acetic acid (4:5:1, by vol., upper phase) as the developing phase. The first fraction eluted is the desired product. After removal of solvent under reduced pressure the residue :5 obtained is dissolved in methanol (5 mL) and passed through a short column (3.5 x 20 cm) of anion exchange resin (Amberlite IRA 400, chloride form). After removal of solvent under reduced pressure from the eluate, the residue is treated with diethylether and dried under high vacuum to give the product as a violet solid.

s H-NMR: OH (300-Iz, CD3OD): 2.20-2.35 (m, 4 H), 2.40-2.50 (m, 4 H), 2.80 (s, 12 H), 3.05 (4 H. t,3J 7.8, 2 H), 3.25 (s, 12 H), 3.45-3.55 (bs, 4 H), 3. 65 3.75(m,4H),4.30(t,3J4.2Hz,4H),7.40,8,10(2xd,3J 7.5Hz,2x4 H), 8.95,9.45 (2x d,3J 4.2 Hz, 8H), 10.40(s,2H). lo

COMPOUND 9 5,15-bis-[4-(3-Triethylammonio-propyloxy)-phenyl]-porphyrin dichloride N±ON Cl To a solution of Compound 7 (50 ma, 0.068 mmol) in absolute DMF (20 mL) is added triethylamine (4,7 mL, 0.034 mol. 500 eq.). The mixture is stirred at 60 C for 24 h. The solvent is removed under reduced pressure and the residue obtained is dissolved in methanol (5 mL) and filtered o through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After washing with methanol (ca. 1L) the pad is eluted with acetic acid:methanol:water (3:2:1, by vol.). After evaporation of the solvent from the eluted fraction, the raw product obtained is dissolved in methanol (5 mL) and purified by chromatography on a column (2.5 x 40 cm) of Sephadex LI-1-20 eluting with n-butanol:water:acetic acid (4:5:1, by vol., upper phase). The solvents are removed under reduced pressure from appropriate fractions, the residue obtained is dissolved in methanol (5 mL) and the solution is passed through a short column (3.5 x 20 cm) of anion exchange resin (Amberlite IRA 400, chloride form) to yield the product as a violet solid after evaporation of solvent.

IH-NMR: OH (300 - , CD3OD): 1.25 (m, 18H), 2.13 (m, 4H), the signals for CH2NCH2 (16I-I) are in the area 3.00-3.40 as a part of the multiplet covered by the solvent signals, 4.15 (t, 4H,3J = 7.5 Hz), 7.36 (d, 4H,3J = 7.5Hz),8.15(d,4H,3J=7.5Hz),9.05(d,4H,3J=7.5Hz),9.54(d,4H, 3J= 7.5 Hz), 10.45 (s, 2H)

*COMPOUND I O

5,15-bis-[4-(3-Trimethylammonio-propyloxy)-phenyl]-porphyrin dichloride 10\ N, N A solution of Compound 7 (300 ma, 0.41 mmol) in absolute DMF (50 mL) is transferred into a 100 mL autoclave. After addition of trimethylamine (4.5 g), the mixture is stirred at 50 C for 16 h. After evaporation of the solvent, the residue obtained is dissolved in methanol so (5 mL) and the solution is filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After washing with methanol (ca. 1L) the pad is elated with acetic acid:methanol:water (3:2:1, by vol. ).

After evaporation of the solvent from appropriate fractions, the residue obtained is dissolved in methanol (5 mL) and purified by chromatography as on a column (2.5 x 40 cm) of Scphadex LH-20, eluting with n butanol:water:acetic acid (4:5:1, by vol., upper phase). Two fractions are obtained, the frst-eluting of which is the desired product. The solvent is removed under reduced pressure and the residue obtained is redissolved in methanol (5 mL) and the solution is passed through a short column (3.5 x 20 cm) of anion exchange resin (Amberlite IRA 40O, chloride form).

After evaporation of the solvent under reduced pressure, the residue is treated with methanol:diethylether and dried under high vacuum to give the product as a violet solid.

1H-NMR: OH (300Mz, CD3OD): 2.40-2.60 (m, 4 H), 3.30-3.25 (bs, 18 H), 3.753.80 (m, 4 H), 4.40(t,3J 7.5 Hz, 4 PI), 7.40, 8.20 (2 x d,3J 8.5 Hz, 8 H), 9. 05, 9.50(2xd,3J4.5 Hz, 8H), 10.45(s,2H).

Alternative synthesis route for Compound 10 Compound 42 (lOOmg, 0.2mMol, see below) is dissolved and potassium carbonate (230mg 1.7mMol) is suspended in DMF (30mL) and to the vigorously-stirred mixture is added a solution of (1-bromopropyl)- trimethylammonium bromide (350mg, 1.3mMol) in DMF (5mL) so dropwise at 50 C during 30 mins. The mixture is heated for 15h. DMF is removed by rotary evaporation and the residue obtained is dissolved in methanol and the solution is filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After washing with methanol (ca. 1L) the pad is eluted with acetic acid:methanol:water :5 (3:2:1, by vol.). After evaporation of the solvent from appropriate fractions, the residue obtained is dissolved in methanol (5 mL) and purified by chromatography on a column (2.5 x 40 cm) of Sephadex LI-I- 20, elating with n-butanol:water:acetic acid (4:5:1, by vol., upper phase) .

Two fractions are obtained, the first-eluting of which is the desired so product. The solvent is removed under reduced pressure and the residue obtained is redissolved in methanol (5 mL) and the solution is passed through a short column (3.5 x 20 cm) of anion exchange resin (Amberlite IRA 400, chloride form). After evaporation of the solvent under reduced pressure, the residue is treated with methanol:diethylether and dried under high vacuum to give the product as a violet solid.

COMPOUND 1 1 5,15 -his- [3 -(3 -Bromo-propyloxy)-phenyl] -porphyrin To a stirred solution of dipyrrolemethane (1.22 g, 8.2 mmol) and 3-(3bromo-propyloxy)- benzaldehyde (2.06 g, 8.2 mmol) in degassed dichloromethane (2 L), TEA (0.14 mL, 3 mmol) is added dropwise. The solution is stirred at room temperature in the dark for 17 h under argon.

After addition of DDQ (5.4 g, 0.024 mol), the mixture is stirred at room temperature for a further lh. After removal of solvents under reduced pressure, the residue obtained is dissolved in dichloromethane (5 mL) and passed through a column (300 g) of silica (Fluke 60) using so dichloromethane as eluent to give raw product which is treated with dichloromethane:methanol to yield pure material as a violet solid.

1H-NMR: 6} (300Mz, CDC13): -3.20 (2 H. s), 2.40 (quint,3J7.5 Hz, 4 H), 3.65 (t,3J 7.5 Hz, 4 H), 4.25 (t, 33 7.5 Hz, 4 H), 7.20-7.25, 7.60-7.65, 7.75-7.80 (3 x m,8H), 9.05,9.25,(2 xd,3J4.2 Hz, 8H), 10.25(s,2H).

COMPOUND 12 5,15-bis-[3-(3-Trimethylammonio-propyloxy)-phenyl]-porphyrin dichloride IN HN \ + N,O O. N- lo A solution of Compound 11 (400 ma, 0.543 mmol) in DMF (50 mL) is transferred into a 100 mL autoclave. After addition of trimethylamine (6.3g), the mixture is stirred at 50 C for 8 h. After evaporation of the solvent under reduced pressure, the residue obtained is dissolved in methanol (5 mL) and the solution is filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After washing the pad with methanol (ca.lL), elusion with acetic acid:methanol:water (3:2:1, by vol.) affords fractions which, after evaporation of the solvent under reduced pressure, gives a solid residue. This is dissolved in to methanol (5 mL) and purified by chromatography on a column (2.5 x 40 cm) of Sephadex LH-20 eluting with n-butanol:water:acetic acid (4:5:], by vol., upper phase). Two fractions are eluted from the column, the first of which is the desired product. After removal of the solvent under reduced pressure, the residue obtained is dissolved in methanol (5 mL).

s The solution is passed through a short column (3.5 x 20 cm) of anion exchange resin (Amberlite IRA 400, chloride form), the solvent is removed under reduced pressure and the raw product is treated with methanol:diethylether to give a violet solid which is dried under high vacuum.

IH-NMR: OH (300Mz, CD3OD): 2.30-2.35 (m, 4 H), 3.15 (s, 18 H), 3.95-4.05 (m, 4 H), 4.20-4.25 (m, 4 I-I), 7.40-7.45, 7.65-7.70, 7.80-7.85 (3 x m, 8 H), 9.00-9.05, 9.40-9.45,(2 x m, 8 H), 10.40 (m, 2 H).

COMPOUND 13 5,15-bis-(4-lydroxy-phenyl)10,20-bis-(4-undecyloxy-phenyl)-porphyrin

OH

C11H23 C11H23

OH

The third fraction eluted from the column during the chromatographic separation described for the synthesis of Compound 2 is characterized as 5,15-bis-(4-hydroxy-phenyl)- 10,20-bis-(4-undecyloxy-phenyl)-porphyrin HNMR: 6 (300MHz, CDCI3): -2.88 (2 H. s), 0.85 (t, 3J7.5 Hz, 6 H), 1.20-].40 (m, 28 H), 1.55 (br m, 4 H), 1.80 (quint, 317.5 Hz, 4 H), 4.15 (I, 3J 7.5 Hz, 4 H), 6.65, 7.15 (d, 31 8.1 Hz, 8 H), 7.80, 8.00 (d, 3J 8.1 Hz, 8 H), 8.75-8.80 (m, 8 H).

trans-Regioisomer geometry is assigned by 'H-3C-2D-NMR in d-acetic acid.

COMPOUND 14 5,1 0-bis-(4-Hydroxy-phenyl)- 1 5,20-bis-(4-undecyloxy-phenyl)-porphyrin

OH

HOiN HN: - The fourth fraction eluted from the column during the chromatographic separation described for the synthesis of Compound 2 is characterised as 5,1 0-bis-(4-hydroxyphenyl)- 1 5,20-bis-(4-undecyloxy-phenyl)-porphyrin lo 'H-NMR: SH (300MHz, CDC13): -2.80 (2 H. s), 0.90 (t, 3J 7.5 Hz, 6 H), 1.20-1.60 (m, ASH), 1.65(quint,3J7.5Hz,4H),2.00(quint,3J7.5 Hz, 4H), 4.22 (t, 3J 7.5 Hz, 4 H), 7.15 (d, 3J 8.1 Hz, 4 H), 7.25 (d, 3J 8.2 Hz, 4 H), 8.10 (d,3J8.2Hz,4H),8.15(d,3J8.2Hz,4H),8.80-8.9O(m,8H).

cis-Regioisomer geometry is assigned by 'H-3C-2D-NMR in d-acetic acid.

COMPOUND 15 5,10,15-tris-[4-(3-Bromo-propyloxy)-phenyl]-20-(4-undecyloXy-phenyl)porphyrin OBr Br Br 0, Under an argon atmosphere, Compound 2 (200 ma, 0. 24 mmol) is dissolved in absolute DMF (40 mL) in the presence of K2CO3 ( 500 ma) and 1,3-dibromopropane (1.02 mL, 10 mmol). The mixture is heated overnight at 80 C. Work-up is as the procedure given for Compound 2 lo described above. The product is purified by column chromatography on silica gel (Merck 60) elating with hexane:ethyl acetate (5: 1, by vol.).

IH-NMR: SH (300MHz, CDC13): -2.75 (2 H. s), 0.85 (t, 3J7.5 Hz, 3 H), 1.201.45 (m, 14 H), 1.50 (quiet, 3J 7.5 Hz, 2 H), 1.90 (quiet, 3J 7.5 Hz, 2 H), 2. 40 (quint, 3J 7.4 Hz, 6 H), 3.65 (I, 3J 7.4 Hz, 6 H), 4.16 (t, 3J 7.5 Hz, 2 H), 4.25 (t,3J7.5 Hz, 6 II), 7.18-7.20 (m, 8 H), 8.00-8.05 (m, 8 H), 8.75-8. 85 (m, 8 H).

COMPOUNI) 16 5,10,1 5-tris-f 4-(3-Triethylammonio-propyloxy)-phcnyl]-20-(4undecyloxy-phenyl)-porphyrin bichloride < o Compound IS (200 ma, 0.17 mmol) is dissolved in absolute DMF (40 mL) with triethylamine (S mL, 34.5 mmol, 208 eq.). The mixture is heated to 50 C for 48 h. After removal of DMF under vacuum, the lo residue obtained is dissolved in methanol and purified by column chromatography using silica gel (Merck, 60) eluting with methanol:water:acetic acid (2:1:3, by vol.) and then acetic acid:pyridine (1:1, by vol.). Removal of solvent from appropriate fractions under vacuum affords raw product which is dissolved in methanol:aqueous NaCl (IM) (S rnL. l:l, by vol.). The mixture is stirred for 30 mins and filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After washing the pad with methanol (200 mL) it is eluted with methanol:water:acetic acid (2:1:3, by vol.). After evaporation of solvent from appropriate combined fractions, the residue obtained is So dissolved in methanol (2mL) and dichloromethane (S mL) is added dropwise. The precipitated white gel is collected by filtration and the solvent is removed under high vacuum.

IH-NMR: OH (300MHz, CD3OD): 0.90 (t,3J7.5 Hz, 3 H), 1.20-1.45 (m, 43H), 1. 45 1.65 (bs, 2 H), 2.25-2.40 (bs, 6 H), 3.35-3.45 (bs, 24 H), 3.50-3.60 (bs,, 6 H), 4.25 (t,3J 7.5 Hz, 2 14), 4.40-4.45 (bs, 6 H), 7.25-7.40, 8.10-8. 20 (m, 16 H), 8.80-9.10 (bs, 8 H).

COMPOUND 17 5-[4-(3 -Hydroxy-phenyl)]- 15-(3 -undecyloxy-phenyl)-porphyrin --O)OH 5-15-bis-(3-Hydroxy-phenyl)-porphyrin (Wiehe, A., Simonenko, E. J., Senge, M. O. and Roeder, B. Journal of Porphyrins and Phthalocyanines 5, 758-761 (2001)) (86 ma, 0.17 mmol) is dissolved and K2CO3 (250 ma, 7.1 mmol) is suspended in DMF (40 mL). To the vigorously-stirred mixture a solution of 1-bromoundecane (0.04 mL, 0.17 mmol) in DMF (5 mL) is added dropwise at 50 C during 30 mins and the mixture is heated at that temperature for 1 h. After removal by filtration of K2CO3, DMF is removed under high vacuum. The residue obtained is purified by column o chromatography using silica gel (Merck 60) eluting with n-hexane:ethyl acetate (10:1, by vol.). The 2nd fraction is collected and dried under high vacuum to give the product.

1H-NMR: 6 (300Mz, CDCI3): -3.15 (2 H. s), 0.75 (t,3J7.5 Hz, 3 H), 1.10-1.30 (m, 14 H), 1.35 (m, 2 H), 1.80 (quiet, 3J 7.5 Hz, 2 H), 4.05 (t,3.J 7.5 Liz, 2 H), 6.85-6.90, 7.20-7.25, 7.35-7.45, 7.50-7.65, 7.75-7.80 (5 x m, 8 H), 8.85, 8.95,9.10,9.20(4xd,3J4.9Hz,4x2H), 10.15(s,2H).

COMPOUND 18 5, ] 0,15-tris-(3-Hydroxy-phenyl)-20-(3-dodecyloxy-phenyl)-porphyrin l0 3Hydroxybenzaldetyde (1.8 g, 14.8 mmol, 3 eqv.) and 3 dodecyloxybenzaldehyde (1.35 g, 4.9 mmol, 1 eqv.) are dissolved in a mixture of acetic acid (145 mL) and nitrobenzene (98 mL, 960 mmol) and heated to 120 C. Pyrrole (1.35 mL, 19.6 mmol, 4 eqv.) is added in one portion and the mixture is stirred at 120 C for lh. After cooling to room temperature, solvents are removed in vacuo at 50 C. The product is isolated by chromatography on a column (500 g) of silica using toluene as eluent. The desired product is obtained as the fifth fraction from the column and is re-chromatographed using a smaller (200 g) silica coulmn eluted with toluene. The product is obtained as a violet solid after evaporation ofthe solvent.

H-NMR: & (300 MHz, CDCI3): 0.64 (t, 3 H,3J6.8 Hz), 0.94-1.15 (m, 16 H), 1.25 (bs, 2 H), 1.62 (bs, 2 H), 3.90 (bs, 2 H), 6.33-6.95 (m, 8 H), 7.08-7.60 (m, 8 H), 8.20-8.47 (m, 4 H), 8.51-8.70 (m, 4 H) COMPOUND 19 5{3-[bis-(2-Diethylamino-ethyl)-aminopropyloxy]-phenyl} - 15 -(3 undecyloxy-phenyl)-porphyrin 03{?O--No Compound 17 (50 ma, 0.065 mmol) is dissolved with N,N,N,N tetraethyldiethylenetriamine (lmL, 39 mmol) in THF(10 mL) and the mixture is stirred at room temperature for 4 days. After evaporation of lo the solvent, the residue is dissolved in diethyl ether (20mL) and the solution is washed with water (5 x 30 mL). The organic phase is dried (Na2SO4) and concentrated under high vacuum. The mixture is purified by column chromatography (silica gel, Merck 60) eluting with n hexane:ethyl acetate (5:1, by vol.) followed byn-hexane:ethyl acetate:triethyl amine (10:10:1, by vol.). After collection of appropriate fractions and removal of solvent under reduced pressure, pure product is obtained by treatment of the residue with diethyl ether:methanol.

H-NMR: b (300Mz, CDC13): 0.80 (t, 3J 7.5 Hz, 3 H), 0.9 (t, 3J 7.5 Hz, 12 TI), 1.20-1.40 (m, 14 H), 1.45 (quint,3J7.5 Hz, 2 H),1.80 (quint,3J7.5 Hz, 2 H) , 1.95 (quint, 3J 7.5 Hz, 2 H),2.40-2.60 (m, 16 H), 2.65 (t,3J 7.5 Hz, 2 H), 4.10 (t,3J7.5 Hz, 2 H), 4.20 (t,3J7.5 Hz, 2 H), 7.30-7.40, 7.55-7.65, 7.75-7.80 (3 x m, 8 H), 9.10-9.15, 9.20-9.25 (2 x m, 2 x 4 H), 10.15 (s, 2 H).

COMPO1; AND 20 5-[4-(3-Bromo-propyloxy)-phenyl]-15-(4-dodecyloxy-phenyl)-porphyrin 00 To a stirred solution of dipyrrolemethane (0.31 g, 2.1 mmol), 4-(3-bromo proyloxy)-benzaldehyde (0.27 g, 1.1 mmol) and 4-dodecyloxy benzaldehyde (0.32 g, 1.1 mmol) in degassed dichloromethane (500 mL).

TEA (0.035 mL, 1.5 mmol) is added dropwise. The solution is stirred at room temperature in the dark for 17 h under argon. After addition of DDQ (1.38 g, 6 mmol), the mixture is stirred at room temperature for a further hour. Purification by column chromatography using silica gel (Merck 60, 400 g) with toluene as eluent affords the product (2n fraction) together with Compound 7 (art fraction).

1H-NMR: SH (300Mz, CDC13): -3.15 (2 H. s), 0.90 (t,3J7.5 Hz, 3 H), 1.20-1. 40 (m, 16 H), 1.55 (quint, 3J 7.5 Hz, 2 H), 1.90 (quint, 3J 7.5 Hz, 2 H), 2.40 (quint, 3J 7.5Hz, 2H), 3.75 (t, 3J 7.5 Hz, 2 H), 4.20 (t, 3J 7.5 Hz, 2 H), 4.35 (t, 3J7.5 Hz, 2 H), 7.20-7.30, 8.10-8.15 (2 x m, 8 H), 9.10-9.15, 9.25-9.30(2 xm,2 x4H), 10.20(s,2H).

COMPOUND 21 5,10,15,20-tetrakis-(3-Hydroxy-phenyl)-porphyrin 3-Hydroxybenzaldehyde (0.910 g, 7.45 mmol) is dissolved in propionic acid (50 mL) and heated to 140 C. Pyrrole (0.52 mL, 7.45 mmol) is added in one portion and the mixture heated at reflux for 2h. Stirring is continued for an additional 12 h at room temperature. Propionic acid is removed in vacuo and the residue dissolved in acetone and purified by chromatography on a column (250 g) of silica which is eluted with toluene containing a continuously increasing proportion of ethyl acetate.

The product is eluted with toluene:ethyl acetate (6:1 by vol.). Solvent is removed in vacuo to afford the product as a violet solid.

IH-NMR: OH (300 Adz, d6-acetone): 7.18 (d, 4H, 3J= 8.25 Hz), 7.49 (t, 4H, 3J= 8.25 Hz), 7.56-7.62 (m, 8H), 8.81 (m, 8 H) COMPOUND 22 5,10,15-tris-[4-(3-Bromo-propyloxy)-phenyl]-20-(4-dodecyloxy-phenyl) porphyrin OBr >0< Or Or C12H25 To a stirred solution of pyrrole (0.7 ml, ] 0 mmol), 4-(3-bromoproyloxy) benzaldehyde (1.8 g, 7.5 mmol) and 4-(ndodecyloxy)-benzaldehyde so (0.725 g, 2.5 mmol) in degassed dichloromethane (1 L) is added TEA (0.085 ml, 10 mmol) dropwise. The reaction solution is stirred under argon at room temperature in the dark for 17 h. After addition of DDQ (6.9 g, 30 mmol), the reaction mixture is stirred at room temperature for a further lh. The solvents are removed under reduced pressure and the :5 residue re-dissolved in toluene. Chromatographic purification on a column (3.5 x 30 cm) of silica gel (Merck 60) using toluene:n-hexane (1:4 by vol.) as eluent gives crude product which is purified by treatment with methanol:dichloromethane, giving a violet solid.

IH-NMR: OH (300MHz, CDC13): 0.90 (t,3J7.5 Hz, 3 H), 1.20-1.45 (m, 16 H), 1.60 (quiet, 3J 7.5 Hz, 2 H), 1.90 (quiet, 3J 7.5 Hz, 2 H), 2.50 (quiet, 3J 7. 4 Hz, 6 H), 3.75 (t,317.4 Hz, 6 H), 4.20 (t,3J7.5 Hz, 2 H), 4.35 (t,317.5 Hz, 6 H), 7.25-7.30 (m, 8 H), 8.15-8.30 (m, 8 H), 8.80-8.85 (m, 8 H).

COMPOUND 23 5- {4-3-Dimethyl-(3 -dimethylaminopropyl)-ammoniopropyloxy]phenyl}- 15-(4-dodecyloxy-phenyl)-porphyrin chloride

-N

C12H25 O Compound 20 (30 ma, 0.038 mmol) is dissolved with N,N,N',N'tetramethyl-1,3-propanediamine (156 ma, 1.2 mmol) in THF:DMF(1:1 by vol., 20 mL) and stirred at 50 C for 18 h. After evaporation of the o solvent under reduced pressure, the residue is dissolved in dichloromethane and purified by column chromatography (silica gel Merck 60) eluting with acetic acid:methanol:water (3:2:1, by vol.). After combining appropriate fractions and removal of solvent under reduced pressure, the residue is treatment with dichloromethane:hexane to afford the product as a violet solid.

1H-NMR: SH (300W, CDC13+1 % acetic acid): 0.85 (m, 3 H), 1.20-1.40 (m, 18 H), 1.55-1.60 (m, 2 H), 1.60-1.65 (m, 4H), 2.10-2.20 (bs, 8 H), 3.15-3.25 (m, 8 H), 3.75 (bs, 2 H), 4.20 (bs, 2 H), 4.35 (bs, 2 H), 7.15-7.20, 8.10-8. 15 (2 x m, 8 H), 8.95-9.00, 9.10-9.15, 9.25-9.30 (3 x bs, 8 H), 10.20 (s, 2H).

COMPOUND 24 5,15-bis-(3 -Methoxy-phenyl)- 10-undecyl-porphyrin

Q

MeO OMe ]O Into a 50 mL flask containing lithium (500 ma, 71 mmol) is added freshly distilled diethyl ether (15 mL) under an argon atmosphere. The suspension is refluxed for l hour, cooled to 15 C and treated with a solution of n-undecylbromide (6.58 g, 71 mmol) in ether (6 mL) added dropwise via syringe. The mixture is cooled to 7-10 C and, after 5 min. when the suspension becomes slightly cloudy and bright spots appear on the lithium metal, the remainder of the n-undecylbromide solution is added at an even rate over a period of 30 min while the internal o temperature is maintained at below 10 C. Upon completion of addition, the mixture is stirred further for 1 h at 10 C. The suspension is filtered under argon to remove excess lithium and lithium bromide.

5, l S-bis-(3 -Methoxy-phenyl)-porphyrin (100 ma, 0.19 mmol) is s dissolved in anhydrous THF (30 mL) at -50 C under an argon atmosphere. The organolithium reagent described above (5 mL) is added dropwise to the mixture. After 5 min the cooling bath is removed and the mixture is warmed to room temperature. After stirring at room temperature for 15 min the reaction is quenched by slow addition of water (2 mL). After IS min the mixture is oxidized by the addition of DDQ (4 mL, 0.4 mmol, 0.1 M in THE) and stirred for a further IS min. The mixture is filtered through alumina (neutral, Brockman grade +) and purified by column chromatography on silica gel eluting with hexane:dichloromethane (4:1 by vol.). The first fraction is collected and treated with methanol:dichloromethane to give a solid product.

SHE R: OH (300Mz, CDC13): -3.05 (bs, 2 H. s), 0.80 (t,3J7.5 Hz, 3 H), 1. 10-1.20 (m, 12 H), 1.25 (m, 2 H), 1.70 (quiet, 3J 7.5 Hz, 2 H), 2.40 (quiet, 3J 7. 5 Hz, 2 H), 3.85 (s, 6H), 4.95 (t,3J7.5 Hz, 2 H), 7.20-7.23, 7.50-7.60, 7. 65 7.75 (3x m, 8 H), 8.85-8.90, 9.10-9.15, 9.35-9.40 (3 x m, 8 H), 9.95 (s, 1H).

COMPOUND 25 3-[({3-[(3-{4-[15-(4-Dodecyloxy-phenyl)-porphyrin-5-yl]-phenoxy}propyl)-dimethyl-ammonio] -propyl} -dimethyl-ammoni o)-propyl] trimethyl-amnonium bichloride 2,N-NO C12H25 O,N Compound 23 (20 ma, 0.022 mmol) and (1-bromopropyl)-trimethyl- ammonium bromide (26 ma, 0.1 mmol) are dissolved in DMF(15 ml) and :5 stirred overnight at 50 C. After evaporation of the solvent under reduced pressure, the residue is dissolved in methanol (5 ml) and applied to a pad (3 cm deep) of silica gel which is washed with methanol (500 ml) followed by acetic acid:methanol:water (3:2:1 by vol.). After evaporation of the solvent the residue is purified by column chromatography (silica gel Merck 60) using at first acetic acid:methanol:water (3:2:1 by vol.) and then pyridine:acetic acid (1:1 by vol.). The second fraction eluted is collected and dried under vacuum. The residue is dissolved in methanol (2 ml) and purified by chromatography on a column (2.5 x 40 cm) of Sephadex LH-20 which is eluted with n-butanol:acetic acid:water (5:1:4 by vol., upper phase). After removal of solvent under reduced pressure, the residue is dried under vacuum at 80 C. NMR spectroscopy indicates lo the product is contaminated with a small proportion of elimination products.

COMPOUND 26 5,10, 1 5-tris-[4-(3-Diethylamino-propyloxy)-phenyl]-20-(4-dodecyloxy phenyl)-porphyrin O--NEt2 NEt2 NEt2 OC12H25 Compound 22 (50 ma, 0.06 mmol) and freshly distilled diethylamine (5 so ml) are dissolved in absolute DMF (30 ml) under argon. The reaction mixture is stirred at room temperature for 20 h and poured into ethyl acetate (50 ml). The mixture is washed with water (4 x 50 ml) and, after drying the combined organic phases (Na2SO4), evaporation of solvent affords a residue which is purified by chromatography on a column (2.5 x :5 30 cm) of silica (Merck 60) which is eluted with ethyl acetate:n- hexane:triethyl amine (10: 10: 1, by vol.). Fractions are combined as appropriate, the solvent evaporated under reduced pressure and the residue dried under high vacuum. Treatment with dichloromethane:n- hexane affords pure product.

H-NMR: 6 (300MlIz, CDCI3): 0.85 (t,3J7.5 I-Iz, 3 H), 1.05 (m, 18 H), 1.20-1.45 (m, 18 H), 1.55 (quiet, 3J 7.5 Hz, 2 H), 2.15 (quiet, 3J 7.5 Hz, 6 H), 2. 75 (quint,, 3J 7.4 Hz, 6 H), 3.15-3.25 (m, 12 H), 4.15 (I, 3J 7.5 Hz, 2 H), 4.25 (t,3J7.5 Hz, 6 H), 7.15-7.20 (m, 8 H), 8.00-8.05 (m, 8 H), 7.95-8.05 lo (m, 8 H) COMPOUND 27 5,15-bis-(3-Hydroxy-phenyl)10-undecyl-porphyrin A, IN Nit

HO JI OH

To a solution of Compound 24 (95 ma, 0.14 mmol) in anhydrous dichloromethane (80 mL) under an argon atmosphere BBr3, (6 mL, 1M in dichloromethane) is added dropwise at -70 C and the mixture is stirred for 1 h. The mixture is warmed to room temperature and stirred overnight then cooled to -10 C and hydrolysed by addition of 2 mL water during 1 h. NaHCO3 (3 g) is added directly to neutralization. The mixture is stirred for a further 12 h. After removal of NaHCO3 by filtration and of dichoromethane under vacuum, the residue obtained is purified by column chromatography using silica gel eluting with dichloromethane. After removal of solvent from appropriate combined fractions and drying under high vacuum the product is obtained as a violet solid.

1H-NMR: OH (300, CDCl3): -3.05 (bs, 2 H. s), 0.85 (t,3J7.5 Hz, 3 H), 1.201.40 (m, 12 Id), 1.50 (m, 2 H), 1.80 (quint,3J7.5 Hz, 2 IT), 2.55 (quint, 3J7. 5 Hz, 2 H), 5.00 (t,3J7.5 Hz, 2 H), 7.15-7.25, 7.50-7.60, 7.80-7.90 (3x m, 8 H), 8.95-9.00, 9.20-9.25, 9.50-9.60 (3 x m, 8 H), 10.15 (s, 1H).

COMPOUND 28 5,15-bis-[3-(3-Trimethylammmonio-propyloxy)-phenyl]10-undecyl o porphyrin dichloride C11H23 _N,N To a solution of Compound 27 (50 ma, 0. 08 mmol) in DMF (20 mL) under an argon atmosphere K2CO3 (100 ma, 0.72 mmol) and (3 bromopropyl)-trimethylammonium bromide (300 ma, 1.2 mmol) are added and the mixture is stirred at 50 C for 18 h. After removal of solvent under high vacuum the residue obtained is dissolved in methanol (5 mL) and filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After washing the pad with methanol (500 mL) it is eluted with acetic acid:methanol:water (3:2:1, v:v). After drying of appropriate combined fractions under high vacuum the residue is dissolved in methanol and purified by column chromatography on Sephadex LH-20 eluting with n-butanol:acetic acid:water (5:1:4, by vol., :5 upper phase). After evaporation of solvent the residue obtained from the first fraction eluted is dissolved in methanol and passed through a short column of anion exchange resin (Amberlite IRA 400, chloride form) to give, after evaporation of solvent, the pure product.

IH-NMR: OH (300Mz, CD3OD): 0.85 (I, 3J 7.5 Hz, 3 H), 1.20-1.40 (m, 12 H), 1.50 (m, 2 I-I), 1.80 (m, 2 H), 2.40 (bs, 4 H), 2.55 (m, 2 H), 3.20 (bs, 18 H), 3.65 (bs, 4 H), 4.35 (bs, 4 H), 5.10 (m, 2 H), 7.50-7.55, 7.70-7.85 (2 x m, 8 H), 8.95-9.00, 9.25-9.24, 9.50-9.70 (3 x bs, 8 H), 10.15 (bs, 1H).

lo COMPOUND 29 5,10-bis-[4-(3-Trimethylammonio-propyloxy)-phenyl]15,20-bis-(4 undecyloxy-phenyl)-porphyrin dichloride

O-N_

_N_ 0}0'' 0.

Compound 14 (50 ma, 0.05 mmol) is dissolved and K2CO3 (150 ma, 1.1 mmol) is suspended in DMF (30 mL). To the vigorously-stirred mixture a solution of (1-bromopropyl)-trimethy]ammonium bromide (0.3 g, 16.6 mmol) in DMF (10 mL) is added dropwise at 50 C and the mixture is o heated for 18 h. After removal of DMF under high vacuum, the residue obtained is dissolved in methanol (5 mL) and filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After washing the pad with methanol (ca. 500 mL) it is eluted with acetic acid:methanol:water (3:2:1, by vol.). After evaporation of solvent from appropriate combined fractions the residue obtained is purified by chromatography on a column (2.5 x 40 cm) of Sephadex LH-20 eluting with n-butanol:water:acetic acid (5:4:], by vol., upper phase) for further separation from the excess ammonium salt and other by-products. After removal of solvent under reduced pressure the residue obtained is dissolved in methanol and passed through a short column (3.5 x 20 cm) of anion exchange resin (Amberlite IRA 400, chloride form). After evaporation of solvent under reduced pressure, the product is dried under high vacuum.

H-NMR: SH (300MHz, CD3OD): 0.80 (t, 3J 7.5 Hz, 6 H), 1.15-1.35 (m, 28 H), 1.35-1.45 (bs, 4 H), 1.70-1.80 (bs, 4 H), 2.30-2.40 (bs, 4 H), 3.15-3.30 (bs, 18 H), 3.65-3.75 (bs, 4 H), 4.00-4.05 (m, 4 H), 4.30-4.40 (bs, 4 H), 7.00-7.15, 7.20-7.30, 7.80-95, 7.95-8.15 (4 x m, 4 x 4 H), 8.60-9.00 (bs, 8 H).

COMPOUND 30 5,10,15-tris-(3-Hydroxy-phenyl)-20-(3-undecyloxy-phenyl)-porphyrin

POOH

HO>3OH 0, _ Pyrrole (1.31 g, 19.6 mmol) is added in one portion to a mixture of 3- hydroxybenzaldehyde (1.8 g, 14.8 mmol) and 3-undecyloxybenzaldehyde (1.36 g, 4.9 mmol) in acetic acid (145 mL) and nitrobcnzene (118 g, 960 mmol) preheated to 130 C and the mixture is stirred for 1 hour at 120 C.

:5 The mixture is cooled and solvent removed under high vacuum. The residue is dissolved in dichloromethane (5 mL) and purified by column chromatography using silica gel (Merck 60) eluting with hexane:toluene (4:1, by vol.). The product is obtained after removal of solvent from the eluate under reduced pressure and drying the obtained residue under vacuum.

IH-NMR: OH (300Mz, CDC13): 0.75-0.80 (m, 3 H), 1.05-1.35 (m, 14 H), 1.401.50 (m, 2 H), 1.75-1.85 (m, 2 H), 3.90-4.10 (m,2 H), 6.90- 7.70 (m, 16 H), 8.45-8.80 (m, 8 H).

COMPOUND 31 5- t4-[3 -Dimethyl-(3 -trimethylammonio-propyl)-ammonio-propyloxy] phenyl} 15-(4-dodecyloxy-phenyl)-porphyrin dichloride

NH

C'2H2s,i,wON IN Compound 23 (50 ma, 0.055 mmol) is dissolved with methyl iodide (5 mL, 80 mmol) in absolute DMF(30 mL) and the mixture is stirred at 40 C for 3h. After evaporation of solvent the residue obtained is dissolved in no methanol (5 mL) and filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After washing the pad with methanol (ca. 1 L) it is eluted with dichloromethane:methanol (2:3 by vol., 500 mL) and then acetic acid:water:methanol (3:1:2, by vol.). After removal of solvent from appropriate pooled fractions the residue obtained iS dissolved in acetic acid and purified by column chromatography on Sephadex LH-20 eluting with acetic acid. After evaporation of solvent from appropriate pooled fractions and drying the residue obtained under high vacuum, the residue is dissolved in methanol and passed through a small column (3.5 x 20 cm) of anion exchange resin (Amberlite IRA 400, chloride form). After evaporation of solvent from the eluate, the product is dried under high vacuum.

COMPOUND 32 5-[4-(3-Dimethyldecyl-ammoniopropyloxy)-phenyl]- l 5{4-[3-dimethyl- (3-dimethylaminopropyl)-ammoniopropyloxy]-phenyl} -porphyrin dichloride

O - RIO--N-C H

--N /

I OMe2N Compound 23 (50 ma, 0.068 mmol) is dissolved with N,N,N',N' tetramethyl-1,3-propanediamine (354 ma, 1.36 mmol) and N,N dimethyldecylamine (1 g, 2.72 mmol) in DMF:THF(30 mL, 1:1, by vol.) Is and the mixture is stirred at 50 C overnight. After evaporation of the solvent under reduced pressure the residue obtained is dissolved in methanol (10 mL) and filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After washing the pad with methanol (ca. 500 mL) it is eluted with acetic acid:methanol:water (3:2:1, so by vol.). The first two fractions eluted are combined and after evaporation of the solvent under reduced pressure the residue obtained is dissolved in methanol and purified by chromatography on a column (2.5 x 40 cm) of Sephadex LH-20 eluting with n-butanol:water:acetic acid (4:5:1, by vol.). After removal of solvent under reduced pressure from as the second fraction eluted, the residue is dissolved in methanol (5 ILL) and passed through a short column (3.5 x 20 cm) of anion exchange resin (Amberlite IRA 400, chloride form). The eluate is evaporated to dryness and the residue obtained is dried under high vacuum to afford the product.

H-NMR: OH (300z, CD3OD): 0.80 (m, 3 H), 1.05-1.25 (m, 10 H), 1.25-1.40 (bs, 2 H), 1.80-1.90 (bs, 4 H), 2.15-2.30 (bs, 2 H), 2.80-3.60 (m, 20 H), 3.80 3.95 (bs, 4 H), 7.05-7.15, 7.85-8.00 (2 x m, 2 x 4 H), 8.75-8.90, 9.20-9. 35 (2xbs,2x4H), 10.15 (bs,2H). lo

COMPOUND 33 5,10,15-tris[3-(3-Trimethyl-ammoniopropyloxy)-phenyl]-20-(3undecyloxy-phenyl)-porphyrin bichloride 4.

_N 0 No

O O

O. _,N Compound 30 (100 ma, 0.12 mmol) is dissolved and K2CO3 (230 ma, 1. 7 mmol) is suspended in DMF (30 mL). To the vigorously-stirred mixture a solution of (1-bromopropyl)-trimethylammonium bromide (0.3 g, 16.6 so mmol) in DMF (10 mL) is added dropwise at 50 C during 30 mins and the mixture is heated for 18 h. After removal of DMF under reduced pressure, the residue obtained is dissolved in methanol (5 mL) and filtered through a pad of silica gel (depth 2 cm) supported on a steel frit (diameter 3.5 cm). After washing the pad with methanol (ca. 500 mL) it :5 is eluted with acetic acid:methanol:water (3:2:1, by vol.). After evaporation of solvent from appropriate combined fractions under reduced pressure, the residue is purified by chromatography on a column (2.5 x 40 cm) of Sephadex LH-20 elating with n-butanol:water:acetic acid (5:4:1, by vol., upper phase). Aider removal of solvent under reduced pressure from the eluate, the residue obtained is dissolved in methanol and the solution is passed through a short column (3.5 x 20 cm) of anion exchange resin (Amberlite IRA 400, chloride form). Evaporation of solvent from the eluate gives the product which is dried under high vacuum.

1H-NMR: SH (300WIz, CD3OD): 0.75-0.80 (m, 3 H), 1.00-1.40 (m, 18 H), 1.601.80 (bs, 2 H), 2.25-2.40 (bs, 6 H), 3.29 (bs, 27 H), 3.40-3.60 (m, 6 H), 3.90-4.00 (m, 2 H), 4.05-4.25 (m, 6 H), 7.10-7.20, 7.25-7.40, 7.60-7.80, 7.80-7.90 (4 x m, 16H), 8.70-9.00 (bs, 8 H).

COMPOUND 34 5,15-bis-(3-Hydroxy-phenyl)-porphyrin HO0OH This is prepared as described by Wiehe, A., Simonenko, E. J., Senge, M. O. and Roeder, B. Journal of Porphyrins and Phthalocyanines 5, 758-761 (2001).

COMPOUND 35 5,10,15-tris-(4-Hydroxy-phenyl)-20-(4-tetradecyloxy-phenyl)-porphyrin

OH

HO{:oCi4H29

OH

5,10,15,20-tetrakis-(4-Hydroxy-phenyl)-porphyrin (170 ma, 0.25 mmol) is dissolved and K2CO3 (0.65 g, mmol) is suspended in DMF (30 mL).

To the vigorously stirred reaction mixture a solution of 1 bromotetradecane (0.1 mL, 0.45 mmol) in DMF (10 mL) is added lo dropwise at 50 C during 30 mins and the mixture is heated for 1.5h.

After evaporation of solvent, the residue is dissolved in toluene:ethanol (1:1 by vol., cat 5 mL) and purified by chromatography using a column (5 x 25 cm) of silica gel (Merck 60) which is washed with toluene. After the elusion of the first 3 fractions, elusion is continued using toluene:ethyl is acetate (2:1 by vol.). The fifth compound eluted is collected, the solvent evaporated and the residue dried under high vacuum to afford product as a violet solid.

H-NMR: & (300MHz, d6-acetone): 0.85 (t,3J7.5 Hz, 3 H), 1.15-1.55 (m, 20 H), 1.45 (quint, 3J 7.5 Hz, 2 H), 1.75 (quint, 3J 7.5 Hz, 2 H), 4.10 (t, 3J 7. 5 Hz, 2 H), 7.20 (d, 3J 8.5 Hz, 2 H), 7.25 (d, 3J 8.5 Hz, 6 H), 8.00-8.15 (m, 8 H), 8.80-9.10 (m, 8 H).

COMPOUND 36 5,10,15-tris-[4-(3-Trimethyl-ammoniopropyloxy)-phenyl]-20-(4tetradecyloxy-phenyl)-porphyrin bichloride of\ 1+ t00 / b I., - The n-tetradecyloxy-analogue of Compound 2, prepared similarly as described above for Compound 2 but using 1-bromotetradecane in place of 1bromoundecane, (50 ma, 0.057 mmol) and (1-bromopropyl) o trimethylammonium bromide (2] 0 ma, 0.8 mmol) are dissolved and K2CO3 (230 ma, 1.7 mmol) is suspended in DMF (20 mL). The vigorously stirred mixture is stirred at this temperature for 18 h. After removal of DMF under reduced pressure the residue obtained is dissolved in methanol (5 mL) and filtered through a pad of silica gel (depth 2 cm) Is supported on a steel frit (diameter 3.5 cm). After washing the pad with methanol (ca. 500 mL) it is eluted with acetic acid:methanol:water (3:2:1, by vol.). After evaporation of the solvent from appropriately combined fractions, the residue obtained is purified by chromatography on a column (2.5 x 40 cm) of Sephadex LH-20 eluting with n-butanol:water:acetic acid so (4:5:1, by vol., upper phase) for separation from the excess of ammonium salt and other contaminating materials. After elusion and removal of the solvent from appropriate fractions, the residue obtained is dissolved in methanol (5 mL) and passed through a short column (3.5 x 20 cm) of anion exchange resin (Amberlite IRA 400, chloride form). Solvent is removed under reduced pressure and the residue obtained is dried under high vacuum to afford the product as a violet solid.

IH-NMR: OH (300MHz, CD3OD): 0.75 (t, 3J 7.5 Hz, 3 H), 0.95-1.25 (m, 22 lI) , ].50-1.65 (bs, 2 H), 2.20-2.40 (bs, 6 H), 3.05-3.15 (bs, 27 H), 3.45-3.60 (bs, 6 H), 3.60-3.80 (bs, 2 H), 4.05-4.25 (bs, 6 H), 6.80-7.25, 7.65-8.05, (2 x m, 16 H), 8.45-8.95 (bs, 8 H).

lo COMPOUND 37 5-(4-{3-[2,4,6-tris-(Dimethylaminomethyl)-phenyloxy]-propyloxy} phenyl)15-(4-dodecyloxy-phenyl)-porphyrin H2sC,20 it- i- O iMe20 _NMe2 Me2N Compound 20 (50 ma, 0.063 mmol) is dissolved in DMF (20 mL) in the presence of 2,4,6-tris-(dimethylaminomethyl)-phenol (1 mL, 3.7 mmol) and stirred at 50 C overnight. After evaporation of the solvent, the residue is solidified by treatment of the residue with dichloromethane:methanol to remove the excess of amine. After filtration, the porphyrins are redissolved in dichloromethane and purified by chromatography on a column of silica gel (Merck 60) which is washed with dichloromethane. Evaporation of solvent under reduced pressure and treatment of the residue with dichloromethane:methanol gives the product as a violet solid.

IH-NMR: OH (300Mz, CDC13): -3.15 (2 H. s), 0.85 (t,3J4.5 Hz, 3 H), 1.20-1. 40 (m, 18 H), 1.55 (quint, 3J 4.5 Hz, 2 H), 1.90 (quiet, 3J 4.5 Hz, 2 14), 2.20 (s, 18H),2.55(t,3J5.2Hz,2H), 3.45 (s,6H),4.15 (t,3J5.5Hz,2H), 4.20 s (t,3J 5. 5 Hz, 2 H), 4.35 (t, 3J 7.5 Hz, 2 H), 6.85 (2 x s, 2 H), 7.20-7.30, 8.10-8.15 (2 x m, 8 H), 9.00-9.05, 9.25-9.30 (2 x m, 2 x 4 H), 10.20 (s, 2 I-I).

COMPOUND 38 l o 5,10,15 -tris-(4-Hydroxy-phenyl)-20-(4-decyloxy-phenyl)porphyrin

OH

H0 C10H21

OH

5,10,15,20-tetrakis-(4-Hydroxy-phenyl)-porphyrin (100 ma, 0.15 mmol) is dissolved and K2CO3 (230 ma) is suspended in DMF (30 mL). To the vigorously stirred reaction mixture a solution of 1-bromodecane (0.016 mL, 0.11 mmol) in DMF (10 mL) is added dropwise at 70 C during 30 mins and the mixture is stirred for 1.5h. Afler evaporation of solvent, the residue is dissolved in toluene:ethanol (1:1 by vol., ca.3 mL) and purified by chromatography on a column (150 g) of silica gel (Merck 60) using toluene as eluent. Afler elusion of the first 3 fractions, the column is eluted with toluene:ethyl acetate (2: 1 by vol.) and the 50' fraction eluted is collected7 the solvent removed and the residue dried under high vacuum to give the product as a violet solid.

1H-NMR: 6 (300Mz, d6-acetone): 0.95 (t, 3J 7.5 Hz, 3 H), 1.25-1.55 (m, 12 H), 1.55 (quint, 3J7.5 Hz, 2 H), 1.85 (quint, 3J7.5 Hz, 2 H), 4.15 (t, 3J7.5 Hz, 2 H), 7.20 (d, 3J 8.5 Hz, 2 H), 7.25 (d, 3J 8.5 Hz, 6 H), 8.00-8.15 (m, 8 H), 8.80-9.10 (m, 8 H).

COMPOUND 39 5,10,15-tris-[4-(3-l rimethylammonio-propyloxy)-phenyl]-20-(4decyloxy-phenyl)-porphyrin bichloride lo owing CI lcl- b t0:0 / Cl Compound 38 (50 ma, 0.061 mmol) and (1-bromopropyl) trimethylammonium bromide (210 ma, 0.8 mmol) are dissolved and K2CO3 (230 ma, 1.7 mmol) is suspended in DMF (20 mL). The vigorously stirred reaction mixture is heated at 50 C for 18 h. After evaporation of solvent, the raw product is dissolved in methanol and purified by chromatography on a column (2.5 x 40 cm) of Sephadex, eluting with n-butanol:water:acetic acid (4:5:1, by vol., upper phase).

o After removal of the solvent, the residue is dissolved in methanol and passed through a column (3.5 x 20 cm) of Amberlite IRA-400 (chloride form) . After evaporation of solvent, the product is dried under high vacuum and yields a violet solid.

H-NMR: 6-1 (300MHz, CD3OD): 0.90 (t, 3J 7.5 Hz, 3 H), 1.20-1.40 (m, 12 H), 1.45-1.60 (bs, 2 H), 1.80-1.90 (bs, 2 H), 2.45-2.55 (bs, 6 H), 3.25-3.35 (bs, 27 H), 3.75-3.85 (bs,, 6 H), 4.05-4.25 (m, 2 H), 4.35-4.40 (bs, 6 H), 7. ] 0-7.40, 7.95-8.15 (2 x m, 16 H), 8.60-9.00 (bs, 8 H).

COMPOIJNO 40 5,10,15-tris-(4-Hydroxy-phenyl)-20-(4-tridecyloxy-phenyl)-porphyrin

OH

HOOC13H27

OH

5,10,15,20-tetrakis-(4-Hydroxy-phenyl)-porphyrin (400 ma, 0.59 mmol) is dissolved and K2CO3 (1.0 g, 7.1 mmol) is suspended in DMF (75 mL).

To the vigorously stirred reaction mixture a solution of 1-bromotridecane (0.1 mL, 0.45 mmol) in DMF (10 mL) is added dropwise at 50 C during mins and the mixture is then heated for 1.5h. The reaction mixture is cooled to room temperature and poured into water (150 mL). The porphyrins are extracted with ethyl acetate (100 mL) and the extract washed with brine (3 x 50 mL) and dried (Na2SO4). After evaporation of o solvent, the residue is dissolved in toluene:ethanol (1:1, by vol., cat 10 mlL) and purified by chromatography using a column (200g) of silica gel (Merck 60) with toluene as the eluent. After the elusion of the first three compounds, the eluent is changed to toluene:ethyl acetate (2:1, by vol.).

The fifth compound eluted is collected and dried under high vacuum to :5 yield product as a violet solid.

]H-NMR: OH (300Mz, d6-acetone): 0.85 (I, 3J 7.5 Hz, 3 H), 1.20-1.60 (m, 18 H), 1.50 (quint, 3J 7.5 Hz, 2 H), 1.80 (quint, 3J 7.5 Hz, 2 I-I), 4.14 (I, 3J 7.5 Hz, 2 H), 7.20 (d, 3J 8.5 IIz, 2 11), 7.25 (d, 3J 8.5 Hz, 6 H), 8.00- 8.15 (m, 8H), 8.80-9.10(m, 8H).

COMPOUND 41 5-(4-Tridecyloxy-phenyl)10,15,20-tris-[4-(3-trimethylammonio- propyloxy)-phenyl]-porphyrin bichloride own\ ' N_} an/-Nix 0,x Compound 40 (50 ma, 0.057 mmol) and (l-bromopropyl) trimethylammonium bromide (210 ma, 0.8 mmol) are dissolved and K2CO3 (230 sing, 1.7 mmol) is suspended in DM1F (20 mL). The vigorously stirred reaction mixture is heated at 50 C for 18 h. After removal of DMF, the residue is dissolved in methanol (5mL) and applied to a pad (2 cm thick) of silica gel which is washed with methanol (ca. 1000 mL) end then eluted with acetic acid:methanol:water (3:2:1 by vol.).

so After evaporation of the solvent the residue is dissolved in methanol and further purified by chromatography on a column (2.5 x 40 cm) of Sephadex LH-20 which is eluted with n-butanol:water:acetic acid (4:5:1 by vol., upper phase). After removal of solvent, the residue is dissolved in methanol and passed through a short column (3.5 x 20 cm) of anion exchange resin (Amberlite IRC 400, chloride form). After evaporation of solvent, the product is dried under high vacuum to afford a violet solid.

1H-NMR: s &n (300MHz, CD3OD): 0.90 (t, 3J 7.5 Hz, 3 H), 1.20-1.40 (m, 18 H), 1.45-1.60 (m, 2 H), 1.80-1.90 (bs, 2 H), 2.40-2.55 (bs, 6 H), 3.25-3.35 (bs, 27 H), 3.75-3.85 (bs, 6 H), 4.05-4.25 (m, 2 H), 4.35-4.40 (bs, 6 H), 7.10-7.40, 7.90-8.15 (2 x m, 16 H), 8.60-9.00 (bs, 8 H).

lo COMPOUND 42 5,15-bis-(4-Hydroxy-phenyl)-porphyrin

HORN HNWOH

is This is prepared as described by Mehta, Gover&an; Muthusamy, Sengodagounder; Maiya, Bhaskar G.; Arounaguiri, S., J.Chem.Soc.Perkin Trans.!; 2177-2182 (1999).

COMPOUND 43 5,10,15-tris-(4-Hydroxy-phenyl)-20-(4-octyloxy-phenyl)-porphyrin o

HO OH

OH

5,10,15,20-tetrakis-(4-Hydroxy-phenyl)-porphyrin (200 ma, 0.294 mmol) isdissolved and potassium carbonate (487 ma, 3.53 mmol, 12 eqv.) is suspended under argon in absolute DMF (50 mL) and the mixture is heated to 55 C. A solution of octyl bromide (35.81, 0.206 mmol, 0.7 eqv.) in absolute DMF (10 mL) is added dropwise during 30 min. and the mixture is stirred at 55 C for 2 h. The solvent is removed in vacuo at 50 C, water (80 mL) is added and the mixture is extracted with ethyl acetate (3 x 40 mL). The combined organic fraction is dried (Na2SO4) lo and the solvent evaporated. The residue is purified by chromatography on a column (300g) of silica gel. Tetra-alkylated and tri-alkylated compounds are eluted with toluene:ethyl acetate (30:1 by vol.). The third fraction (all-substituted compound, trans-isomer) is eluted with toluene:ethylacetate (15:1 by vol.). The fourth fraction (all-substituted Is compound, cis-isomer) is eluted with toluene:ethyl acetate (10:1 by vol.) and the desired product (mono-alkylated compound) is eluted with toluene:ethylacetate (5: 1 by vol.). The solvent is removed under reduced pressure and the residue dried under high vacuum to give the product as a violet solid.

H-NMR: H (300 z, d6-acetone): 0.75 (t, 3H, 3J= 6.8 Hz), 1.13-1.25 (m, 8H), 1.43 (quint, 2H, 3J= 7.5 Hz), 1.73 (quint, 2 H. 3J= 7.5 Hz), 3.50 (t, 2H, 3J = 8 Hz), 7.11 (d, 2H, 3J = 7.5 Hz), 7.16 (d, 6 H. 3J= 7.5 Hz), 7.90-7.94 (m, 8H), 8.80-8.90 (m, 8 H) COMPOUND 44 5-(4-Dodecyloxy-phenyl)10,15,20-tris-(4-hydroxy-phenyl)-porphyrin p HO in{ - OH

OH

5,10,15,20-tetrakis-(4-Hydroxy-phenyl)-porphyrin (200 ma, 0.294 mmol) is dissolved and potassium carbonate (487 ma, 3.53 mmol, 12 eqv.) in suspended under argon in absolute DMF (50 mL) and the mixture is heated to 55 C. A solution of dodecyl bromide (49.41, 0.206 mmol, 0.7 lo eqv.) in absolute DMF (10 mL) is added dropwise during 30 min. The mixture is stirred at 55 C for 2 h. The solvent is removed in vacuo at 50 C, water (80 mL) is added and the mixture extracted with ethyl acetate (3 x 40 mL). The combined organic fractions are dried (Na2SO4) and the solvent evaporated. The product is isolated by chromatography on a column (300g) of silica. Tetra-alkylated and tri-alkylated compounds are eluted with toluene:ethyl acetate (30:1 by vol.), all-substituted compound (trans-isomer) with toluene:ethyl acetate (15:1 by vol.), all-substituted compound (cis-isomer) with toluene:ethyl acetate (10:1 by vol.) and the desired product (mono-alkylated compound) with toluene:ethyl acetate o (5:1 by vol). Solvent is removed in vacuo and the residue dried at high vacuum to give product as a violet solid.

IH-NMR: 6-1 (300 MHz, d6-acetone): 0.75 (t, 3H, 3J= 6.8 Hz), 1.13-1.25 (m, 16H), :5 1.41 (quint, 2H, 3J= 7.5 Hz), 1.63 (quint, 2 H. 3J= 7.5 Hz), 3.89 (t, 2H, 3J = 6 Hz), 7.11 (d, 2H, 3J = 7.5 Hz), 7.16 (d, 6H, 3J= 7.5 Hz), 7.9-7.94 (m, 8H), 8.78-8,83 (m, 8 H) COMPOUND 45 s 5,10,15-tris-(4-Hydroxy-phenyl)-20- (4-nonyloxy-phenyl)-porphyrin o Ho30H

OH

5,10,15,20-tetrakis-(4-Hydroxy-phenyl)-porphyrin (200 ma, 0.294 mmol) lo is dissolved and potassium carbonate (487 ma, 3.53 mmol, 12 eqv.) is suspended under argon in absolute DMF (50 mL) and the mixture heated to 55 C. A solution of nonyl bromide (49.4111, 0.206 mmol, 0.7 eqv.) in absolute DMF (10 mL) is added dropwise during 30 min. The mixture is stirred at 55 C for 2 h. The solvent is removed in vacua at 50 C, water (80 mL) is added and the mixture extracted with ethyl acetate (3 x 40 mL). The combined organic extracts are dried (Na2SO4) and solvent removed under reduced pressure. The product is isolated by chromatography on a column (300g) of silica. Tetra-alkylated and tri alkylated compounds are eluted with toluene:ethyl acetate (30:1 by vol.), So all-substituted compound (trans-isomer) with toluene:ethyl acetate (15:1 by vol.). all- substituted compound (cis-isomer) with toluene:ethyl acetate (10:1 by vol.) and the desired product (mono-alkylated compound) is eluted with toluene:ethyl acetate (5:1 by vol.). The solvent is removed under reduced pressure and the residue dried at high vacuum to afford the :5 product as a violet solid.

]H-NMR: OH (300 Adz, d6-acetone): 0.87 (t, 3H, 3J= 7.5 Hz), 1.]4-1.26 (m, lOH), 1.41 (quint, 2H), 1.70 (quint, 2H, 3J= 7.5 Hz), 3.92 (t, 2H, 3J= 7.5 Hz), 7.02 (d, 2H,3J = 8.25 Hz,), 7.15 (d, 6H, 3J = 7.5 Hz,), 7.85 (d, 2H, 3J = 8.25 Hz), 7.91 (d, 33= 7.5Hz), 8.76-8,84 (m, 8 H) COMPOUND 46 5-(4-Octyloxy-phenyl)- 10,15,20-tris-[4-(3-trimethylammonio o propyloxy)phenyl]-porphyrin bichloride p

FOCI- ;

I Cl Cl Compound 43 (50 ma, 0.063 mmol) and (3-bromopropyl) trimethylammonium bromide (164mg, 0.63 mmol, lOeqv.) are dissolved and potassium carbonate (130 ma, 0.95 mmol, 15 eqv.) is suspended under argon in absolute DM1F (30 mL) and the mixture is stirred at 55 C for 12 h. The solvent is removed in vacuo at 50 C and the residue applied to a pad (2 cm deep) of silica. The unreacted ammonium salts are washed offwith methanol (lOOOmL) and the product is elated with acetic so acid:methanol:water (3:2:1 by vol.). The solvent is removed under reduced pressure and the residue further purified by chromatography on a column (lOOg) of Sephadex LH-20 using n-butanol:water:acetic acid (4:5:1 by vol., upper phase) as the eluent. The solvents are removed under reduced pressure and the residue dissolved in methanol and passed through a small column of anion exchange resin (Amberlite IRA 400, chloride form) using methanol as eluent. After evaporation of solvent, the crude product is dissolved in the minimum amount of methanol and diethylether (50 mL) added. The solution is centrifuged for 15 min. The supernatant liquid is evaporated to dryness and the residue dried at high vacuum to give the product as a violet solid.

IH-NMR: OH (300MHz, CD3OD): 0.90 (t, 3H, 3J= 7.5 Hz), 1.25-1.41 (m, 8H), 1.45 (bs, 2H), 1.87 (bs, 2H), 2.38 (bs, 6H), 3,29 (bs, 27H), 3.67 (t, 6H,3J= 7. 5 Hz), 4.01 (I, 2H,3J= 7.5 Hz), 4.30 (t, 6H, 3J= 7.5 Hz), 7.11 (d, 2H, 3J= 7.5 Hz), 7.38 (d, 6H, 31= 7.5 Hz), 7.95 (d, 2H, 37= 7.5 Hz), 8.11 (d, 6H, 3J= 7.5 Hz), 8.93 (bs, 8H) COMPOUND 47 5-(4-Dodecyloxy-phenyl)- 10,15,20-tris-[4-(3-trimethylammoniopropyloxy)-phenyl]-porphyrin bichloride _

-N C' 0}< Cl Cl

O

Compound 44 (50 ma, 0.059 mmol) and (3-bromopropyl) trimethylammonium bromide (154mg, 0.59 mmol, 10eqv.) are dissolved and potassium carbonate (122 ma, 0.885 mmol, 15 eqv.) is suspended under argon in absolute DMF (30 mI,) and the mixture is stirred at 55 C for 12 h. The solvent is removed in vacuo at 50 C and the residue re dissolved in a little methanol and applied to a pad of silica (2 cm deep).

The unreacted ammonium salts are washed off with methanol ( l OOOmL).

The product is eluted with acetic acid:methanol:water (3:2:1 by vol.).

The solvents are removed under reduced pressure and the crude product further purified by chromatography on a column (l Dog) of Sephadex LHusing n-butanol:water:acetic acid (4:5:1 by vol., upper phase) as eluent. The solvents are removed under reduced pressure, the residue re- dissolved in a little methanol and the solution passed through a short column of anion exchange resin (Amberlite IRC 400, chloride form) lo using methanol as eluent. After removal of solvent the crude product is re-dissolved in the minimum amount of methanol and diethyl ether (50 mL) added. The solution is centrifuged for 15 min. The supernatant liquid is evaporated to dryness and the product dried at high vacuum to give a violet solid.

IH-NMR: 61 (300MHz, C D3O D): 0.88 (t, 3H, 3J = 7.5 Hz), 1.25-1.37 (m, 16H), 1.48 (bs, 2H), 1.93 (bs, 2H), 2.42 (bs, 6H), 3,28 (bs, 27H), 3.68-3.75 (m, 6H), 4.05 (t, 2H), 4.33 (t, 6H), 7.17 (d, 2H, 3J = 7.5 Hz), 7.33 (d, 6H, 3J = o 7.5 Hz), 7.99 (d, 2H, 3J = 7.5 Hz), 8.08 (d, 6H, 3J = 7.5 Hz), 8. 85 (bs, 8H) COMPOUND 48 5-(4-Nonyloxy-phenyl)10,15,20-tris-[4-(3-trimethylammonio- propyloxy)-phenyl]-porphyrin bichloride

O

C

O NO

Compound 45 (50 ma, 0.062 mmol) and (3-bromopropyl) trimethylammonium bromide (162mg, 0.62 mmol, 10eqv.) are dissolved and potassium carbonate (128 ma, 0.93 mmol, 15 eqv.) is suspended lo under argon in absolute DMF (30 mL) and the mixture is stirred at 55 C for 12 h. The solvent is removed in vacuo at 50 C and the residue re dissolved in a little methanol and applied to a pad of silica (2 cm deep).

The unreacted ammonium salts are washed off with methanol (lOOOmL).

The product is eluted with acetic acid:methanol:water (3:2:1 by vol.).

The solvents are removed under reduced pressure and the product further purified by chromatography on a column (lOOg) of Sephadex LH-20 eluting with n-butanol:water:acetic acid (4:5:1 by vol., upper phase). The solvents are removed under reduced pressure, the residue re-dissolved in a little methanol and the solution is passed through a short column of no anion exchange resin (Amberlite IRC 400, chloride form) using methanol as eluent. After removal of solvent, the product is dried at high vacuum to give a violet solid.

1H-NMR: b (300MHz, CD3OD): 0.89 (t, 3H, 3J= 7.5 Hz), 1.18-1.34 (m, lOH), 1.41 (bs, 2H), 1.73 (quint, 2H, 3.J= 7.5 Hz), 2.30-2.44 (m, 6H), 3,31 (bs, 27H), 3.65-3.73 (m, 6H), 3.93 (t, 2H, 3J= 7.5 Hz), 4.25-4.42 (m, 6H), 7.08 (d, 2H,3J= 7.5 Hz), 7.30 (d, 6H,3J= 7.5 Hz), 7.93 (d, 2H, 3J= 7.5 Hz) , 8.05 (d, 6H,3.J= 7.5 Hz), 8.94 (bs, 8H) COMPOUND 49 5-(4-Octyloxy-phenyl)10,15,20-tris-[4-(5-trimethylammonio-pentyloxy) o phenyl]-porphyrin bichloride \ , . C' owl

Cl- Cl 0 Compound 43 (23 ma, 0.03 mmol) and (5-bromopentyl) s trimethylammonium bromide (84 ma, 0.3 mmol, lOeqv.) are dissolved and potassium carbonate (62 ma, 0.45 mmol, 15 eqv.) is suspended under argon in absolute DMF (15 mL) and the mixture is stirred at 55 C for 12 h. The solvent is removed in vacuo at 50 C and the residue re-dissolved in a little methanol and applied to a pad (2 cm deep) of silica. The unreacted ammonium salts are washed offwith methanol (lOOOmL). The product is eluted with acetic acid:methanol:water (3:2:1 by vol.). The solvents are removed under reduced pressure and the product further purified by chromatography on a column (lOOg) of Sephadex LH-20 using n- butanol:water:acetic acid (4:5:1 by vol., upper phase) as eluent.

:5 The solvents are removed under reduced pressure, the residue re dissolved in a little methanol and the solution passed though a short column of anion exchange resin (Amberlite IRC 400, chloride form) with methanol as eluent. The complete purification process is repeated if impurities remain in the product. After removal of solvent, the residue is dried at high vacuum to give the product as a violet solid.

IH-NMlR: OH (300z, CD3OD): 0.78 (bs, 3H), 1.08-1.35 (m, lOH), 1.45-1.59 (m, 6H), 1.63-1.93 (m, 14H), 3.17-3.32 (m, 6H), 3,31 (bs, 33H), 3.84 (bs, 2H), 4.07 (bs, 6H), 6.93 (bs, 2H), 7.09 (d, 2H,3J= 7.5 Hz), 7.74 (bs, 2H), 7.88 (d, 2H,3J= 7.5 Hz), 8.71 (bs, 8H) COMPOUND 50 5,10,15-tris-[4-(5-Trimethylammonio-pentyloxy)-phenyl]-20-(4undecyloxy-phenyl)-porphyrin bichloride \ ,o _N- 9 (00: C- Cl_N\_ ON\ O Compound 2 (50 ma, 0.06 mmol) and (5-bromopentyl) trimethylammonium bromide (174 ma, 0.6 mmol, 10eqv.) are dissolved and potassium carbonate (124 ma, 0.9 mmol, 15 eqv.) is suspended under argon in absolute DMF (30 mL) and the mixture is stirred at 55 C for 12 h. The solvent is removed in vacua at 50 C and the residue re-dissolved in a little methanol and applied to a pad (2 cm deep) of silica. The unreacted ammonium salts are washed off with methanol (lOOOmL). The product is eluted with acetic acid:methanol:water (3:2:1 by vol.).

Solvents are removed under reduced pressure and the product further purified by chromatography on a column (long) of Sephadex LH-20 eluting with n-butanol:water:acetic acid (4:5:1 by vol., upper phase).

Solvents are removed under reduced pressure, the residue re-dissolved in the minimum of methanol and the solution passed through a short column of anion exchange resin (Arnberlite IRC 400) with methanol as eluent.

The complete purification process is repeated if impurities remain in the product. After removal of solvent, the residue is dried at high vacuum to lo give the product as a violet solid.

H-NMR: OH (300MHz, MeOD): 0.71-0,88 (m, 13H), 0.91-1.38 (m, 14H), 1.48 1.81 (m, 12H), signals for -CH2NCH2 and OCH2-long alkyl chain are part of the multiplet together with the signals for solvent in the area 2.8 - 3.3, 3.91 (bs, 6H), 6.33 (bs, 2H), 6.86 (bs, 6H), 7.35 (bs, 2H), 7.70 (bs, 6H), 8.65 (bs, 8H) COMPOUND 51 no 5,10,15,20-tetrakis-(3-Dodecyloxy-phenyl)porphyrin :'_ W] on <: Pyrrole (0.7 mL, 10 mmol) and 3dodecyloxybenzaldehyde (2.9] g, 10 mmol) are dissolved in degassed dichloromethane (1000 mL) and TEA (0.77 mL, 10 mmol) is added dropwise. The mixture is stirred for 17h at room temperature in the dark. DDQ (6.81 g, 30 mmol) is added in one portion and the mixture is stirred for a further lh at room temperature.

The mixture is filtered through a column (400g) of silica using dichloromethane as eluent followed by dichloromethane to which triethylamine is added to adjust the pH value to 8. This purification process is repeated if impurities remain in the product until the pure lo product is obtained.

H-NMR: 6 (300 MHz, d6-acetone): 0.80 (bs, 12H), 1.03-1.45 (m, 80H), 1.78 (quint., 8H, 3J= 7.5 Hz), 4.05 (t, 8H, 3J= 7.5 Hz), 7.24 (d, 4H, 3J= 7.5 Hz), 7.49-7.55 (m, 4H), 7.68-7.71 (m, 8H), 8.80 (m, 8 H) EXAMPLE B: BINDING OF EXEMPLARY COMPOUNDS WITH BACTERIAL

CELLS

Binding of Compounds 8, 10 and 12 wih E. cold E. cold cells were incubated for 5 min with Compound 8, 10 or 12 at various concentrations (1-7.5 M). At the end of the incubation period, :5 the cells were sedimented by centrifugation to remove the fraction of unbound test compound and the cell pellet was resuspended in 2 ml of 2% SDS to obtain cell Iysates. After overnight incubation with SDS, the amount of cell-bound test compound was estimated by spectrofluorimetric analysis of the cell Iysates. The concentration of the so compounds in the cell lysates was calculated by measuring the intensities at the maximum of the emission fluorescence spectrum and interpolating the data on a calibration plot. The amount of cell-bound test compound was expressed as nmoles of compound per mg of cell protein. The protein concentration was determined by the method of Lowry (Lowry et al., 1951, J. Biol. Chem. 193:265-275).

All experiments were run in triplicate and the results represent the average of 3 determinations with standard deviations.

lo The amount of porphyrin recovered from the cells is shown in Table 1.

Table 1

Concentration Bound compound (nmoles/mg cellproteins) of compound (pM) (a) O washings Compound 8 Compound 12 Compound 10 0.01 0.024 + 0.01 0.041 + 0.02 0.026 + 0.005 0.1 0.056 + 0.02 0.151 + 0.02 0.274 + 0.05 0.5 0.522 + 0.2 0.806 + 0.14 1.542 + 0.350 1 3.670 + 0.7 2.70 + 0.30 2.70 + 0.354 (b) 3 washings Compound 8 Compound 12 Compound 10 0.01 0.009 + 0.001 0.021 + 0.005 0.015 + 0.0004 0.1 0.030 + 0.02 0.089 + 0.02 0.078 + 0.02 0.5 0.274 + 0.15 0.622 + 0.10 0.334 + 0.092 1 2.230 + 0.8 1.930 + 0.20 1.278 + 0.102 The results shown in Table I show that the three test compounds bind to E. cold with similar efficiency and that about 50% of the compound that is associated to the cells at the end of the incubation period (5 min) is removed by 3 washings with PBS.

EXAMPLE C: STABILITY STUDIES Chemical stability The following HPLC methodology was established for the analysis of the exemplary compounds of the invention.

The method involves detection by UV at a wavelength of 420 nm, which is very specific for these compounds. In order to monitor lo impurities not related to the porphyrin structure (and therefore not absorbing at 420 nm) UV spectra of the whole chromatograms were also recorded between 200 nm and 700 rim by DAD (diode array detector) in certain experiments.

Column: Zorbax Phenyl, 250 x 4.6 mm, 5 Em Eluent A: 1.5 g sodium dodecylsulfate + 1 mL formic acid in 1000 mL water Eluent B: 1.5 g sodium dodecylsulfate + 1 mL formic acid in 200 mL water + 800 mL :0 tetrahydrofurane Gradient: Time Eluen! B [mind lo/l 0 50 31 65 32 90 33 50 43 50 Flow rate: 0.4 mL/min Detection: 420 nm Column temperature: 25 C Injection volume: 10 pi Solutions: Porphyrin derivatives were dissolved in eluent A to give a final concentration of approximately 0.3 mg/ml.

Typical retention time of the exemplary compounds was approximately 8 minutes (18 minute runtime).

Qualitative stress tests were undertaken on the exemplary compounds of the invention. Analysis was undertaken by HPLC & LC-MS. The compounds were stress tested in solid form, in an aqueous solution and a solution made up in phosphate-buffered saline buffer. The samples were initially incubated for 7 days at 50 C and a sample removed for testing.

The samples were then incubated for a further 7 days at 70 C, samples so removed as before and the samples incubated further for 7 days at 90 C.

HPLC analysis of freshly prepared solutions was undertaken and compared to the samples after 7, 14 and 21 days incubation. A visual comparison of the chromatograms was then undertaken and the content of the main products and by-products as area percentage values determined Is (see Figure 2).

The 3D plots of the chromatograms show no indications for additional formation of fragments (no signals at lower wavelengths) The plot in Figure 3 shows the sample after 21 days in PBS buffer, which showed the largest degradation effect. The results demonstrated minimal degradation on analysis of solid drug and drug in solution heated to 80 C for a number of weeks.

Conclusions

Compounds]0 and 12 were both found to exhibit good stability and were very stable even under the stressed conditions of the test protocol.

lo Although Compound 8 was less stable than Compounds 10 and 12, the stability demonstrated was found to be sufficient for practical use.

Stability of exemplary compounds in formulations The stability of three exemplary compounds (Compounds 8, lO and 12) and one reference compound (Compound 1), stored at 40 C in the dark over 8 weeks in polyethylene vials in various aqueous-based formulations, was evaluated as follows: So - Sodium laureth sulphate (SLES) + water - 9:1 water:ethanol - SLES + 9:1 water:ethanol UV spectra were recorded over the range 350- 700 rim over a period of 7 weeks and a visual evaluation of the samples made at 8 weeks.

The results indicate that all compounds tested exhibited good stability over an eight-week period (see Figure 4).

For Compounds 8 and 10, the stability study was extended to 17 weeks (see Figure 5).

Claims (70)

1. Claims 1. Use of a compound of formula I below in the preparation of a

   medicament for sonodynamic therapy X1 Y41.Z:1 Y3 Y2 X3 wherein: X1, X2, X3 and X4 independently represent a hydrogen atom, a lipophilic moiety, a phenyl group, a lower alkyl, alkaryl or aralkyl group, or a cationic group of the following formula; - L - Rat - Nt(R2)(R3)R4 wherein: L is a linking moiety or is absent; so Rat represents lower alkylene, lower alkenylene or lower alkynylene, which is optionally substituted by one or more substituents selected from lower alkyl, lower alkylene (optionally interrupted with oxygen), fluoro, ORE C(0)R6, C(0)0R7,C(0) HR8R9, NR10R,, and N+R2RI3R14; and R2, R3 and R4 independently represent H. aryl, lower alkyl, lower alkenyl or lower alkynyl, the latter three of which are optionally substituted by one or more substituents selected from lower alkyl, lower alkylene (optionally interrupted with oxygen), aryl, ORs, C(0)R6, C(0)0R7, C(o)NRxR9' NRoR, and N-R2R3R4 Z is -Cll or N; and Y,Y2,Y3 and Y4 are absent or independently represent aryl, lower alkyl, lower alkenyl or lower alkynyl, the latter three of which are optionally substituted by one or more substituents selected from lower alkyl, lower alkylene (optionally interrupted with oxygen), aryl, OR5, C(0)R6, C(0)0R7, C(0)NR8R9, NRioR, and N+R'2R3R4; and R5, R6, R7, Rx' R9, Rlo, Ri R'2, Ri3 and Rj4 independently represent H or lower alkyl provided that at least one of X, X2, X3 and X4 is a cationic group as defined above and at least one of X, X2, X3 and X4 iS a hydrogen atom.
2. 2. Use of a compound of formula II below in the preparation of a medicament for sonodynamic therapy 4Z<1 X4 / M x2 Y3/WY2

   II

   wherein M is a metallic element or a metalloid element and A, X2, X3, X4, Ye, Y2, Y3, Y4 and Z are as defined in Claim 1.

   lo
3. 3. A use according to Claim 2 wherein M is a divalent or trivalent metallic element.
4. 4. A use according to any one of Claim 2 or 3 wherein M is selected from Zn (II), Cu (II), La (III), Lu (III), Y (III), In (III) Cd (II), Mg Is (II), Al(III), Ru, Ni(II), Mn(III), Fe(III) and Pd(lI).
5. 5. A use according to any one of Claims 2 to 4 wherein M is a metalloid element, for example silicon (Si) or germanium (Ge).

   to
6. 6. A use according to any one of the preceding claims wherein Ye, Y2, Y3 and Y4 are absent.

   -
7. 7. A use according to any one of the preceding claims wherein Z is CH.
8. 8. A use according to any one of the preceding claims wherein Rut is an unsubstituted lower alkylene, lower alkenylene or lower alkynylene group.
9. 9. A use according to any one of the preceding claims wherein Ret is (CH2)m- and 'm' is an integer between l and 20.
10. 10. A use according to Claim 9 wherein 'm' is an integer between 1 and lO, for example between 1 and 6, between l and 5, between 1 and 4 or between 1 and 3.

    Is
11. 11. A use according to Claim 10 wherein 'm' is 3.
12. 12. A use according to any one of the preceding claims wherein R2, R3 and/or 1 are lower alkyl, lower alkenyl or lower alkynyl groups.
13. 13. A use according to Claim 12 wherein R2, R3 and/or R4 are
14. 14. A use according to Claim 12 or 13 wherein at least one of R2, R3 and R4 is an alkyl group which is substituted with a primary, secondary or tertiary amine group or a quaternary ammonium group.
15. 15. A use according to any one of the preceding claims wherein Rat is -(CH2)3-, R2 and R3 are CH3 and R4 is - (CH2)3-N(CI-13)2.
16. 16. A use according to any one of the preceding claims wherein Rat is -(CH2)3-, and R2, R3 and R4 are each CH3.
17. 17. A use according to any one of the preceding claims wherein Rat is -(CH2)3-, and R2, R3 and R4 are each C2H5.
18. 18. A use according to any one of the preceding claims wherein L is selected from the group consisting of phenoxy, phenylene, sulfonyl arnido, aminosulfonyl, sulfonylimino, phenylsulfonyl-amido, lo phenylaminosulfonyl, urea, urethane and carbamate linking moieties.
19. 19. A use according to Claim] 8 wherein A, X2, X3 and/or X4 are (OR)n wherein R is -R-N+(R2)(R3)R4, as defined in Claim 1 and 'n' is an integer between 1 and 3.
20. 20. A use according to Claim 18 wherein A, X2, X3 and/or X4 are Rm no wherein R is -R-N+(R2)(R3)R4, as defined in Claim 1 and 'm' is an integer between 1 and 3.
21. 21. A use according to Claim 18 wherein A, X2, X3 and/or X4 are (OR)n \ \/ Rm wherein each R independently is -Row (R2)(R3)R4, as defined in Claim 1 and 'n' and 'm' are integers between l and 3 and wherein the sum of'n' and 'm' is an integer between 1 and 3.
22. 22. A use according to any one of Claims 19 to 21 wherein 'n' or 'm' is3.
23. 23. A use according to any one of Claims 19 to 21 wherein 'n' or 'm' lo is2.
24. 24. A use according to any one of Claims 19 to 21 or 23 wherein 'n' and/or 'm' is 1.
25. 25. A use according to any one of Claims 19 to 21 wherein L is mono
26. 26. A use according to any one of Claims 19 to 21 wherein L is mono or all-substituted at a meta-position(s).
27. 27. A use according to any one of Claims 19 to 21 wherein L is mono or all-substituted at an or/ho-position(s).
28. 28. A use according to any one of the preceding claims wherein the 2s compound comprises two cationic groups, as defined in Claim 1, on opposite sides of the porphyrin ring, i.e. at ring positions 5 and or ring positions l O and 20.
29. 29. A use according to Claim 28 wherein X and X3 are a hydrogen atom, a lipophilic moiety, a phenyl group, a lower alkyl, alkaryl or aralkyl group and X2 and X4 are cationic groups, or vice versa.
30. 30. A use according to any one of Claims 1 to 29 wherein the compound comprises two cationic groups, as defined in Claim 1, on neighbouring positions of the porphyrin ring, i.e. at ring positions 5 and 10, or ring positions 10 and 15, or ring positions 15 lo and 20 or ring positions 20 and 5
31. 31. A use according to Claim 30 wherein X and X2 are hydrogen and X3 and X4 are cationic groups, or X2 and X3 are hydrogen and X4 and X are cationic groups.
32. 32. A use according to any one of the preceding claims wherein at least one of A, X2, X3 and X4 iS a lipophilic moiety.
33. 33. A use according to Claim 32 wherein the lipophilic moiety is a saturated, straight-chain alkyl group of formula - (CH2)pCH3 wherein 'p' is an integer between 1 and 22.
34. 34. A use according to Claim 33 wherein 'p' is between 1 and 18, for example between 2 and 16 or between 4 and 12.
35. 35. A use according to any one of Claims 1 to 31 wherein none of A, X2, X3 and X4 iS a lipophilic moiety.
36. 36. A use according to any one of the preceding claims wherein none of X, X2, X3 and X4 iS a phenyl group.
37. 37. A use according to any one of the preceding claims wherein the compound is water-soluble.
38. 38. A use according to Claim 1 wherein the compound is 5,15-bis-(4 {3-[(3-Dimethylamino-propyl)-dimethyl-ammonio]-propyl-oxy} phenyl)porphyrin dichloride.
39. 39. A use according to Claim 1 wherein the compound is 5,15-bis-[4 o (3Triethylammonio-propyloxy)-phenyl]-porphyrin all-chloride.
40. 40. A use according to Claim 1 wherein the compound is 5,15-bis-[3 (3-Trimethylammonio-propyloxy)-phenyl]-porphyrin dichloride.
41. 41. A use according to Claim 1 wherein the compound is 5,15-bis-[4 (3-Trimethylammonio-propyloxy)-phenyl]-porphyrin dichloride.
42. 42. A use according to Claim 1 wherein the compound is 5-[3,5-bis (3-Trimethylammonio-propyloxy)-phenyl]- 1 5-undecyl-porphyrin dichloride.
43. 43. A use according to Claim I wherein the compound is 5-{4-[3Dimethyl-(3 -dimethylaminopropyl)-ammonio-propyl-oxy] phenyl}-15-(4-dodecyloxy-phenyl)-porphyrin chloride.
44. 44. A use according to Claim 1 wherein the compound is 3-[({3-[(3- {4-[1 5-(4-Dodecyloxy-phenyl)-porphyrin-5-yl]-phenoxy} propyl)-dimethyl-ammonio]-propyl} -dimethyl-ammonio)-propyl] trimethyl-ammonium bichloride.
45. 45. A use according to Claim 1 wherein the compound is S,15-bis-[3(3-Trimethylammmonio-propyloxy)-phenyl]- 1 O-undecyl porphyrin dichloride.
46. 46. A use according to Claim 1 wherein the compound is 5-{4-[3 Dimethyl(3-trimethylammonio-propyl)-ammonio-propyloxy] phenyl}-1 5-(4-dodecyloxyphenyl)-porphyrin dichloride.
47. 47. A use according to Claim 1 wherein the compound is 5-[4-(3 o Dimethyldecyl-ammoniopropyloxy)-phenyl]- 15- {4-[3 -di-methyl (3-dimethylaminopropyl)-ammoniopropyloxy]-phenyl} -porphyrin dichloride.
48. 48. A use as deemed in any one of Claims 38 to 47 wherein the compound is in a metallated form.
49. 49. A use according to any one of the preceding claims wherein the compound is substantially non-toxic to mammalian cells.

    so
50. 50. A use according to any one of the preceding claims wherein the medicament is for killing or attenuating the growth of microorganisms.
51. 51. A use according to Claim 50 wherein the microorganisms are as selected from the group consisting of bacteria, mycoplasmas, yeasts, fungi and viruses.
52. 52. A use according to Claim 50 or 51 wherein the microorganisms are bacteria which are resistant to one or more conventional antibiotic agents.
53. 53. A use according to any one of Claims 50 to 52 wherein the microorganisms are on a light-inaccessible surface or in a light inaccessible area. s
54. 54. A use according to any one of the preceding claims wherein the medicament is for use in the curative and/or prophylactic treatment of microbial infections.

    lo
55. 55. A use according to any one of the preceding claims wherein the medicament is for preventing and/or treating dermatological infection.
56. 56. A use according to any one of the preceding claims wherein the is medicament is for preventing and/or treating an infection of the lungs.
57. 57. A use according to any one of the preceding claims wherein the medicament is for preventing and/or treating wound infection and/or ulcers.
58. 58. A use according to any one of the preceding claims wherein the medicament is for preventing and/or treating cancer.

    :5
59. 59. A method for treating a patient in need of treatment with sonodynamic therapy comprising administering to the patient a compound as described in any one of Claims 1 to 58 and exposing the compound to ultrasound radiation.
60. 60. A method according to Claim 59 wherein the ultrasound radiation has a frequency of about 0.1 to 10 MHz, for example about 0.5 to MHz or about 1 to 3 MHz.
61. 61. A method according to Claim 59 or 60 wherein the ultrasound radiation has a power level (intensity) of about 4 to 12 W/cm2, for example about 2 to 8 W/cm2.
62. 62. A method according to any one of Claims 59 to 61 wherein the lo ultrasound radiation has a duration of about 1 second to 60 minutes, for example about 1 minute to 40 minutes.
63. 63. A method according to any one of Claims 59 to 62 wherein the patient has a microbial infection. ]5
64. 64. A method according to any one of Claims 59 to 62 wherein the patient has a dermatological infection or lung infection.
65. 65. A method according to any one of Claims 59 to 62 wherein the To patient has a wound infection.
66. 66. A method according to any one of Claims 59 to 62 wherein the patient has a cancer.

    :5
67. 67. A method for preventing wound infection comprising contacting the wound with a compound as described in any one of Claims I to 48 and exposing the compound to ultrasound radiation.
68. 68. A method for killing microorganisms in vitro comprising so contacting the microorganisms with a compound as described in any one of Claims 1 to 48 and exposing the compound to ultrasound radiation.
69. 69. Use of a compound in the preparation of a medicament substantially as hereinbefore described with reference to the

    description.
70. 70. A method for killing microorganisms substantially as hereinbefore

    described with reference to the description.