PT103828B - NANOMAGNETE-PORFIRIN HYBRID MATERIALS, PROCESS FOR ITS SYNTHESIS AND THEIR APPLICATION IN FORMULATIONS FOR DISINFECTION OF WATERS - Google Patents

Abstract

The present invention relates to the synthesis of novel NANOMAGNETE-PORFIRIN HYBRID MATERIALS OF GENERAL FORMULA (I) AND TO ITS USE FOR PHOTOINACTIVATING MICRORGANISMS PRESENT IN CONTAMINATED WATERS. THE PROCESS USED FOR THE SYNTHESIS OF THESE HYBRID MATERIALS INVOLVES THE IMMOBILIZATION, THROUGH COVALENT CONNECTION, OF MESO-TETRA-ARILPORFIRINES IN NANOPARTICLES COMPOSED BY A SILIC-COATED MAGNETIC NUCLEUS. NEW NANOMAGNETE-PORFIRIN HYBRID MATERIALS PRESENT GENERAL FORMULA I, IN WHICH: AR1 REPRESENTS 4-TETRAFLUOROPHENYLENE; AR2 = AR3 = AR4 is AR1 -X-R-Z, 4-PYRIDYL, 4-N-METHYLPYRIDINYL AND PHENYL; X is AMINO; R IS A PROPYLENE GROUP; Z ARE MAGNETIC NANOPARTICLES COATED WITH SILICA FUNCTIONALIZED WITH TRIMETHYLAMONIOPROPYLENE GROUPS.

Description

DESCRIPTION

NANOMAGNETE-PORPHYRIN HYBRID MATERIALS, PROCESS FOR THE SUMMARY AND APPLICATION IN WATER DISINFECTION FORMULATIONS

Invention Field

The present invention describes novel nanomagnete-porphyrin hybrid materials, their synthesis as well as the effect these new materials have on the inactivation of microorganisms present in microbiologically contaminated waters.

Background of the invention

Chemically, porphyrins have a base macrocycle composed of four pyrrole rings joined together by metal bridges.

Nanomaterials are characterized by being on a nanometer scale (10 ^-9 m) of one, two or three dimensions. At present, there has been a particular interest in magnetic nanoparticles, namely those made of iron oxide (FesOJ).

Nanomagnete-porphyrin hybrids refer to materials

made up of these two ; systems. The possibility in use porphyrins, as photosensitizers, at the treatment of diseases caused by microorganisms (bacteria, fungi virus and protozoa) is quite promising and it can be a good

alternative to existing antimicrobial agents [1].

In addition, the use of antimicrobial photosensitizers can also be extended to disinfect water, surfaces such as hospital surfaces, etc.

In this case the need for recovery and reuse of the photosensitizer makes it imperative to use supports that meet these requirements [2–6].

Several studies have been reported in the literature showing that it is possible to increase the solubility of the photosensitizer in biological fluids and to increase its selectivity for pathogenic microorganisms with respect to healthy cells, using the immobilization of the compounds, particularly in polymers such as polylysine [7,8].

In the area of catalysis, the concept of recovery and reuse is highly valued since the vast majority of catalysts, such as metalloporphyrins, have a high cost and it is necessary for catalyst separation of reaction product reagents to be easy, feasible and feasible. without loss of activity by the catalyst.

One way to solve this problem is to anchor the catalyst on a solid surface, namely by covalent bonding [9].

The possibility of combining the photoinactivating capacity of porphyrins with the advantages of using a support that allows the reuse of the antimicrobial agent opens the prospect of building new compounds for water disinfection.

Therefore, the choice of an effective porphyrin derivative that allows covalent attachment to a water-insoluble surface (and which can easily be removed from the aqueous medium) is imperative.

As regards porphyrin derivatives that have been shown to be effective in photoinactivating microorganisms, notably in photoinactivating bacteria, cationic derivatives [8] such as 5- (4-carboxyphenyl) -10,15 triiodide stand out. .20-tris (N-methylpyridinium-4yl) porphyrin.

With regard to bacterial activity, there are significant differences between Gram-positive and Gram-negative cells, particularly as regards the constitution of the walls of each of these cell types. These differences dictate the different sensitivity of Gram-positive and Gram-negative to the photoinactivation process.

Neutral photosensitizers, including porphyrins, such as 5- (4-carboxyphenyl) -10,15,20-tri (4pyridyl) porphyrin, and phthalocyanines, effectively destroy Gram-positive bacteria, but do not promote appreciable strain inactivation. Gram-negative bacteria unless the outer membrane permeability of these bacteria is altered before irradiation is initiated. This change in outer membrane permeability is achieved by pretreatment with chemical or biological agents such as CaCl ₂ , EDTA (ethylenediaminethoacetic acid) nonapeptide) [10-13].

or

PBNP [polymyxin

The lower sensitivity of gram-negative bacteria to photoinactivation is due to the presence of the outer membrane, which decreases the permeability to large or hydrophobic compounds compared to common biological membranes [14]. The outer membrane has an asymmetric double structure consisting of an inner layer of phospholipids and an outer layer of lipopolysaccharides (LPS). LPS have negatively charged groups, responsible for negatively charging divalent cation binding to membrane A surface such as Mg ²⁺ , decreasing electrostatic repulsion between

Adjacent LPS, thus playing a key role in the structure and integrity of the outer membrane [15]. In this membrane there are transport proteins, porins, which allow low molecular weight compounds to pass (<600-700 daltons to but prevent high molecular weight compounds to pass through).

The low sensitivity of gram-negative bacteria to photoactivation can be overcome by the use of cationic photosensitizers, which can thus effectively photoinactivate both groups of bacteria [16,17].

Regarding the choice of media, there are a number of requirements to consider to ensure the effectiveness of the photoinactivation process. Proper support should allow the photosensitizer to be accessible and available for the photodynamic process, have functional groups that allow covalent attachment to the photosensitizer preventing water loss, be inert or increase the inactivating capacity of the photosensitizer, and have characteristics allow easy, fast and effective recovery of the photosensitizer.

Silica nanoparticles with magnetic properties can be used as a support, since they allow photosensitizer accessibility, due to: (1) they have dimensions that allow them to enter cells (eg bacterial); (2) these materials may be derivatized with suitable functional groups; (3) Given their magnetic characteristics they can easily be recovered by applying an external magnetic field [18–21].

Magnetic silica nanomaterials are usually made up of a magnetic iron oxide (Fe ₃ O ₄ ) core which is subsequently coated with a silica mesh. Iron oxide nanoparticles can be synthesized by co-precipitation of Fe (II) and Fe (III) salts by addition of base in an oxidizing environment according to equation [22,23]:

Fe ²⁺ + 2Fe ³⁺ + 80H ”-> Fe ₃ O ₄ + 4H ₂ O

The nature of the counterions and the pH of the suspension plays a major role in the stabilization of magnetic particles.

Applications of magnetic fluids, which are stabilized solely by electrostatic repulsion, are restricted as the system is very sensitive to pH variations and has low flexibility in adjusting particle surface properties. The stabilization of magnetic nanoparticles then appears as an important study target since the applicability of this type of nanomaterials depends on the stability [24].

Stabilization of magnetic fluid suspensions can be achieved by following several strategies: by coating the surface of the particles with organic surfactants, using polyethylene glycol polymers, etc. [25.26].

Stabilization can also be achieved by using coating processes, for example with silica. This is one of the most commonly used methodologies since the silica mesh acts as a protective agent for the magnetic core, prevents the formation of large aggregates in liquids and enables further functionalization of surface silanol groups with various coupling agents.

Functionalization, in addition to allowing the binding of other stabilizing agents, also allows the coupling of other molecules for therapeutic purposes [18-21].

There are two main methods for preparing magnetic silica nanoparticles: the Stöber method and the silicic acid method.

In Stöber's method, silica is formed in situ by hydrolysis and condensation of a sol-gel precursor such as tetraalkylorthoxylates.

In the silicic acid method, silicic acid is gradually formed from a solution of a silicate during the addition of an acid that promotes a polymerization reaction. From the silanol (Si-OH) groups siloxane (Si-O-Si) bonds are formed via condensation reaction and the silica coating progressively forms around the magnetic core. Silicic acid molecules are assumed to associate with Fe ₃ O ₄ particles that contain surface -OH groups by forming covalent bonds after condensation. There is evidence that the silicic acid method is the most effective as it allows coating larger surface portions of the magnetic core. [19,2730].

After coating with silica it is possible to derivatize its surface with functional groups that make possible covalent bonding to other compounds. Functionalization with alkoxysilanes is widely used, as there are hydroxyl groups on the surface of the coating which can react, for example, with the aminopropyltriethoxysilane alkoxysilane, allowing further reactions with the amino group of this derivatizing agent [30,31].

In accordance with the present invention, it has been observed

amazing results because involve one effective photosensitizer, broad spectrum in action antimicrobial, and that can be removed someway easy of the aqueous medium to be disinfected. General Description of the Invention The present invention relates to the obtaining in new

nanomagnete-porphyrin hybrid materials, the process

X ambience of attentive to s i.new, the expression lo '' materials' · designs: all oomgos Replacements by uaooparti calas eu tycoons st ..:. ica · testo; <.: ar · ® ea <.ws sis; idle fields pranuirsores of cured ma o o r i a s s r i b r 1 a Nitrides

rotoram ss materi.aiS: stitsldos polos uíwws: w omagnete and pors ira na.

riovos hibrioos materials, and magnetic narwpartloulat porphyrinogenic irlnas general formula _f. I, et pat:

Ar ⁴

Ar ^<;

Λι ^Λ represents 4-tetrafluorophenylene;

Ar ^z - .p; t; pp ^; Ar ³ '- X - R-Zf 4-pyridyl, wworylpyridyl and feuyl;

X is ww

R is a propylene group;

These are magnetic nanoparticles coated with furyanalicated silica with trimethylammonopropylene groups.

In accordance with the invention, particular mention is made of materials 3, 3a, 5, 7, 8, 10, 11.

The above materials have the following chemical structures:

• η

X

?

Rb material 8

X ν ν _{F Γ} i Ν '' \ · bd b- <rMNbCHbi

A ~ j j / v

hybrid materials are obtained after immobilization of meso-tetraarylphyrids sm. magnetically-coated nanoparticles of silica derivative which preferably have the following groups:

Ar ⁵ Ari Ar ', At X £ ϊ z ; Z.v F F .................................. F F : ............. ..................... ; 10:,: im <im » : S10 ·· dubSlab: u? M I ..... / d ..... \; ™ -ς F Ϊ s-i —M Ç ,, y w »R ......:.B..... _Ci , g. yv> —V «Sssz ..., / d .... FF FF w d VR ⁵ Nu-CO-Nu ” • -o4chg :: R _? d ' i hjdroxidquiki, .. i hímOXicsómmslqmk ·,: ρ »: ί>;» Οί ^; 0§ :: Κΰί,; feíiopírnsf:; »» qmí - <> 1 o » no XZ : 'o':? j \ í ύ ^: 31ίϊΰ ': <Χνίί? · ίί <íu!? <> i i í í i i

R ⁵ '-'CHy - (CH2ÚCH3, - [. (X: h _? € h ₂ hgh

In the context of. In the present invention, a variety of cationic and cationic laughter hybrids are tested in oracetic-like strains. 001: 101-000001000 a (in terms of the Ibaoteriophobic somatic samplings of Isohsrichia coli).

Some preliminary trials of photuinacti were also made. using decal bacteria present in ischemic wastewater and faecal entoocholes as a model), often used as indicators of focal pollution.

The best results were obtained only with cationic hybrids and inhibitions of about 10%.

baked, gresenue zztsrtao tmnouaura that the nz.Czzuus uauomagn ste-purtzrlne ohm eoozvloaue arrtzmlcro.ozana may be used the ones in the disenchantment of resilient eagles.

s? ‘« teh newcomer ué z uz na muar, hen reasperaso maquét i has

X. Preparation of shapho-nanoparticles:

In the synthesis of the non-hybrid hybrid materials and porphyrous nanoparticles described in the present invention, nanoparticles coated with the deritized silica were used. with aminopropyl groups.

For support purposes, silica nanoparticles with magnetic properties have been chosen since they allow the photosensitizer to be accessible and available. For the photodynamic process, it has functional groups (amluopropyl groups) that allow covalent attachment to the photosensitizer preventing its loss in water, which may increase the Inactivating capacity of the photosensitizer.

thus, this material was prepared in 3 steps x a; (b) silica coating and (c) functionalization using one. alkoxy-silica according to the following chemical transitions:

The preparation of the iron core is carried out using the co-pre-precipitation method of Fe (1) and Fe (111): by the addition of an aqueous ammonia solution which provided for the rapid formation of a / 11 oxide precipitate. and Fe (1H).

Ror fia · iisn-opartíoslas are derlrati sodas: using Ιο .4; '; Op.0000 40tOXr -: 50.4 3.00: .4: 8, 4 4'4 4 Λ:. 0; 'Ά 003 000 3

SUpS.D.I, 404S from 06008 000000 00 eXVStSX; OOOpOO 60000X4 Id, OStCX undergo reactions like the frog, which is ambiguous. 300:.

2.

Repair of purfixins are affected by meso-superstitious start-up symmetric [Ar ^: ··· - r - Ar '- Ara symmetrical dog · Α' ^{, Λ} ? Arç arç, selected by presenting pentafí.torofenile gopes, which allow covalest connection to the magnetic support.

The symmetric starting porphyrins were prepared by pyrrole condensation with the appropriate aldehyde asthma xtissura, reflex acid / lyclobenene acid, according to methods described in the literature. These conditions allow the tetromerlation of pyrrole and aldehyde, followed by oxidation of the macroel.

For non-symmetrical weeks, they were prepared in a similar way, but in aldehyde we used 4-year old sma mmtum, in relative proportions, it was possible to

The maerootolor; They were solved by aprtom-ntatom in me. mars buy pentaf lacvrafso.i joc eme permto..totom a lloefeo camaleais ae> toparia mstotoiem fòntomaslltodo .. tornlm οροοοse- emaa Acti.sse ct ptofinoins mas-scbatit sidas slmtoremas a s ηοΙοόΡ>; females in m.m the premamento of the mother in maas grspas near flaorofatolo went to eat all iodas ,. Etoe Ramp On

am kc a n i s.mc from s ub to i e. u to p

good astermic derivatives, top of the ptotof istotoer group.:. io _; have been topped and recited if 4pitoto.to; The latter may later be removed after the cationotocation of the myocloclc to take over the remaining positions kk of the ptorocycle.

I have the form was possible. obtain the sixteenth porfirisas:

1) 5,1Q, 15,10-tmtrapto, a (pentafinoropathol> porafine) (2)

11) 5-pent.afluoroferd 1-10,15,20-tr1,4-p.rld.l 1) by fyrin (4)

1.11: 5.10, Itotropha atoto-toto ipsntaf Inorophethol} perfyrin. (9} d poto.ir.ina.aato.dto.to 5-iptotaf Itorofanl trl-lodet) 10,10? ais ((ttomtoi.lp.irid lato'4-1)} from the (6) site, taken from 5-pentaflosorophate 10,1a, 2'-tri.r 4. ~ ρ1 r xdll? by irisa 04 i.

\ A <'Â X J ac acer 1 cgia 1 before the

3. Separation of materials No; P. 11 ·

i hibxxdos nano & agnete-porphysine:

st ϊ nas

SC.2ÇÍ2 · i d i chlor obéo z ·: - ?. η.ο was cc J. çn t ç so ´ and action: a o

; ·;:. to t .; :.:.: 2 according to

added ume.

material overuse

At this time the reacoaonal seam was washed crude with olorofermol (solvent where the porphyrin gives to Ida and sollvelò, and measures to remove the broken porphyria _: unreacted. Sata p.roosg.imanto is terminated when Per iv-Via no longer observed Panda Soren, the porphyrin sterilizer, in the washout solvent, so that all unreacted porphyrin was removed.

The resulting material 3, unlike the material 1 which was brown in color, had a reddish color, thus confirming the presence of immobilized porphyrin.

C ^: Materials 5, 7 and 10 (made up of nanopar magnetic facets coated with aminopropyl groups, some of which are covalently bonded to 5- (4-thiofluorophenylene) porphyryl radicals -10.15 21-tris (4-pyridyl) porphyryl;

- (4 -t e t r a f 1 o r o f 'n 11 e η o) ~ 10,15,2 0 - t ri s i 4 -methylpyridinium) porphyrinyl and? ·· (4-tetra.fluorophenylene) ··

11.15, 20-triphenyl-porphyrinyl, respectively) were obtained by an analogous procedure to that described for material 3, with two caveats: in the case of compounds 7 and 10, the reaction occurred in. dimethyl sulfoxide (DMSG), but is most suitable for the dissolution of porphyrins, and the washings of the obtained compounds were reallocated with chloroform / methanol (37: 2; .a in the case of compound 7 to the ether of the methylene chloride (7: 3) in the case of compound 10.

The compounds; are suspended in bare DMF? Reaction reaction. to which methyl lobate is subsequently added. The reaction mixture. and allowed to stir for about 16 h at 4 ^0: Finally, this time is washed with acetone and water to remove excess methyl iodide (as this is an extremely toxic compound which, if not removed, becomes a hazardous contaminant).

The adapted methylation procedure aimed to cationate free primary amino groups that did not react with porphyrin derivatives.

The presence of charge in these groups increases the repulsion between nearby nanoparticles, preventing the formation of aggregates observed in the suspensions of their precursors in water. In this way, materials 3a, S and 11 are obtained.

4. Measurement of the amount of immobilized porphyrin:

The determination, by UV-Vis spectrophotometry, of the amount of porphyrin in the washing solvent, allowed to calculate, by difference, the amount of immobilized porphyrin.

All synthesized immobilized derivatives were characterized by UV-Vis solids. In the solid UVVis spectra, it is possible to observe the appearance of the Soret band characteristic of porphyrin precursors in nanomagnete-porphyrin hybrids, confirming that these new materials contain immobilized porphyrin photosensitizer.

From scanning electron microscopy (SEM) and transmission electron microscopy (TEM) images made for starting material 1 (aminopropyl functionalized magnetic silica nanoparticles) it is possible to observe, inter alia by TEM, that the size of nanoparticles is on average less than 20 nm. In terms of electron microscopy, no significant differences were found between this material and the resulting hybrids obtained, indicating that after the immobilization process the materials retain their shape and size.

The selected area electron diffraction (SAED) image, which relates to material 1, indicates that the magnetic particles are crystalline. This information is important because the magnetic properties of the material depend on the type of magnetic core structure (Fe ₂ O ₃ ).

The existence of crystalline structure is responsible for an improvement in its magnetic characteristics.

5. Biological Assays

In order to determine the efficacy of the novel compounds synthesized according to the present invention in water treatment, biological tests were carried out using two bacterial strains recognized as a major contaminant in such environments, as well as the effectiveness of inactivating the compounds in bacteria in wastewater samples. Virus inactivation efficacy tests were also performed.

The effectiveness of photosensitizers was evaluated in microcosms. A microcosm is an experimental setup that aims to mimic an event that under normal circumstances should occur naturally but is intended to be reproduced in a controlled environment. In the present case the microcosm represents a 600 ml beaker set containing sample of microorganisms (pure cultures of bacteria and viruses or contaminated wastewater) with a 2.7 cm high solution column. To these samples different concentrations of photosensitizers (in free or immobilized form) were added and white light irradiation under stirring at 25 ° C for 270 minutes.

Then, the degree of photoinhibition at different irradiation times produced by the tested compounds and materials was evaluated.

5.1. Photoinactivation of collector bacterial strains

Two collector bacterial strains were used, one Gram-negative and one Gram-positive, respectively Escherichia coli ATCC 13706 (USA) and Enterococcus faecalis ATCC 51299 (USA) to test the efficacy of the new compounds on these two types.

The photoinactivating efficacy of the free porphyrin precursor 6 in said E. coli and E. faecalis strains at different concentrations was first evaluated for comparison with the results of the hybrid compounds synthesized according to the present invention.

An analogous procedure was then adopted for derivatives 7, 8 and 11, but using higher concentrations (20 and 200 μΜ) of photosensitizer since in this case the compounds were not in solution and would be less available for photoinactivation.

First, the time required for total photoinactivation of the strains under study was determined.

Thus, samples of Escherichia coli and Enterococcus faecalis collection cultures, mentioned above, containing photosensitizers (6, 7, 8 and 11) at different concentrations were irradiated with white light for 60, 90, 180 and / or 270 minutes under stirring at 25 ° C.

Three controls were performed: dark control with photosensitizer (control 1), dark control without photosensitizer (control 2) and control without photosensitizer but exposed to the same irradiation procedure (control 3).

The effects of neutral material 1 (aminopropyl-functionalised silica magnetic nanoparticles) and cationic la (trimethylamoniopropyl-functionalised silica magnetic nanoparticles) on bacterial samples subjected to the same irradiation conditions and in the dark were also studied. photosensitizers.

Bacterial content in the different samples was evaluated by counting colonies after sowing by incorporation in TSA medium. By determining the bacterial content, it is possible to know, by comparison with the initial number of bacteria present in each microcosm, the reduction in the bacterial population caused by the addition of photosensitizer.

Thus, the following results were obtained in terms of bacterial photoinactivation time:

Table 1- Irradiation time (min) after which total photoinactivation of Escherichia coli and Enterococcus faecalis strains (10 ⁴ CFU mL ^-1 ) for porphyrin 6 and

for materials 7, 8 and 11. Concentration (μΜ) Photosensitizer 0.5 1 5th 20 200 6th 180 180 90 - - 7th - - - * * 8th - - - 180 90 11 - - - 180 90

* - There was no significant photoinactivation until

270 minutes of irradiation

For E. coli total photoinactivation was observed after 90 or 180 minutes, depending on the concentration and porphyrin used (Table 1). For E. faecalis the photoinactivation was found to be total for the same irradiation times.

For all cases the time required for total photoinactivation of the Escherichia coli strain was first studied, and it was also found that at these irradiation times the photoinactivation of the Enterococcus faecalis strain was total. This procedure was adopted knowing that Gram-negative strains are much less sensitive to photoinactivation than Gram-5.2 strains.

Photoinactivation of bacteria in natural wastewater samples

In the wastewater samples were analyzed two groups of bacteria that are often used as indicators of fecal pollution: a group of Gramnegative bacteria, fecal coliforms (CF), and a group of Gram-positive bacteria, fecal enterococci (EF). .

CF and EF contents were determined by counting colonies on filtration membranes.

The results of bacterial inhibition in natural wastewater samples are shown in Table 2. Assays were repeated in independent experiments at least twice.

Wastewater samples containing photosensitizers 6, 7, 8 and 11 at different concentrations were irradiated with white light for 50,

90, 180 and / or 270 minutes while stirring at 25 ° C.

First, the photoinactivating efficacy of precursor 6 in Gram-negative bacteria (indicated by the presence of CF, 10 ⁴ CFU mL ' ¹ ) was evaluated using the 5 μΜ concentration, as this was the most effective concentration to inactivate the Escherichia coli strain previously studied and results are given in topic 5.1.

The minimum time required for total CF photoinactivation was also 90 minutes as observed for the collection strain.

Total inactivation of PE (10 ⁴ CFU mL ¹ ) was also observed after 90 minutes for derivative 6.

An analogous procedure was adopted for derivatives 7, 8 and 11, but using higher concentrations of material-bound photosensitizer (20 and 200 μΜ). As for free porphyrin 6, the time required for total CF photoinactivation was first evaluated. Subsequently, it was found that at these irradiation times the photoinactivation of PE was also quite high (> 97%).

Table 2 - Irradiation time (min) at the end 0.0 which shows total photoinactivation of CF and EF strains (10 ⁴ CFU / mL) for porphyrin 6 and derivatives 7, 8 and 11.

Concentration (μΜ) Photosensitizer 5th 20 200 6th 90 - - 7th - * 8th - + 180 11 - + 180

* - No significant photoinactivation up to 270 minutes of irradiation.

- Photoinactivation greater than 99.0%

5.3. Virus Photoinactivation

The results of somatic coliphage inhibition produced from Escherichia coli (ATCC 137003), evaluated by the methods described in the experimental part of the biological assays, are presented in Tables 3. The assays were repeated in independent experiments at least twice.

Somatic coliphage samples containing different concentrations of photosensitizers were established, which were subsequently irradiated with white light for 60, 90, 180 and / or 270 minutes under agitation at 25 ° C.

The photoinactivating efficacy of precursor 6 in somatic coliphages (10 ⁷ PFU mL ' ¹ ) produced from Escherichia coli was also performed at 5 μΜ (concentration at which the shortest time needed for total photoinactivation of the bacterial strain Escherichia coli) was observed.

The minimum time required for total photoinactivation of the somatic coliphages was also 90 minutes as observed for Escherichia coli, Enterococcus faecalis, CF and EF (Table 3).

Under the same conditions, but using higher photosensitizer concentrations (20 and 100 μΜ), derivatives 7, 8 and 11 showed total photoinactivation at irradiation times below 90 minutes, however observed for bacteria.

Table 3 - Irradiation time (min) after which total photoinactivation of somatic coliphages of

Escherichia coli (10 ⁷ PFU / mL) following action of derivatives 6,

7, 8 and 11

Concentration (μΜ) Photosensitizer 5th 20 100 6th 90 - - 7th - 60 60 8th - 30 30 11 - 60 30

Brief Description of the Figures

The figures show the scanning electron microscopy (SEM) and transmission electron microscopy (TEM) images of the starting material 1.

Figure 1: SEM images of material 1;

Figures 2 and 3: TEM images of material 1;

Figure 4: SAED images of material 1

From scanning electron microscopy (SEM) and transmission electron microscopy (TEM) images it can be seen that the size of nanoparticles is on average less than 20 nm. In addition to size, it is also possible to observe the shape of the particles and to compare starting materials with their hybrid derivatives.

The selected area electron diffraction (SAED) image indicates that the magnetic particles are crystalline. The existence of crystalline structure is responsible for an improvement in its magnetic characteristics.

Detailed Description of the Invention

I - Synthesis of the new nanomagnetic porphyrin compounds

- Preparation of magnetic nanoparticles:

Magnetic nanoparticles coated with aminopropyl derivatized silica were used as follows:

1.1. Iron oxide core synthesis (Fe3O ₄ ) - Iron core preparation is performed using the method of co-precipitation of Fe (II) and Fe (III) salts by addition of base (aqueous ammonia solution).

1.2. Silica Coating - Silica coating was performed using the silicic acid method using sodium metasilicate as the source of silicon oxide.

1.3. Functionalization using an alkoxy silane - Nanoparticles are derivatized using 3-aminopropylethoxy silane according to the methodology described in [19,30,31].

- Preparation of porphyrins:

Symmetrical (Ar ¹ = Ar ² = Ar ³ = Ar ⁴ ) and non-symmetrical (Ar ¹ Φ, Ar ² , Ar ³ , Ar ⁴ ) starting substituted porphyrins were used.

2.1. Symmetric starting porphyrins were prepared by condensing pyrrol with the appropriate aldehyde in a refluxing acetic acid / nitrobenzene mixture according to methods described in the literature [32].

2.2. Non-symmetrical porphyrins were prepared analogously, but instead of an aldehyde a mixture of aldehydes was used.

2.3. Cationic porphyrin 6 was synthesized from porphyrin 4. This process was performed using methyl iodide in DMF according to the procedure described in the literature [8]. After precipitation of the reaction mixture with ethyl ether, the product was filtered and recrystallized from a water / acetone mixture.

- Preparation of hybrid materials - Immobilization of porphyrins in magnetic nanoparticles

The newly prepared hybrid materials result from the coupling of various porphyrinic macrocycles to magnetic silica nanoparticles derivatized with aminopropyl groups or trimethylamoniopropyl groups. From the various porphyrins and nanomaterials previously prepared, the new nanomagnetepcrfirin hybrids are prepared, and then the methylation of the hybrid materials is made to obtain materials containing trimethylamoniopropyl groups.

3.1. Compound 3 was synthesized by coupling porphyrin 2 to material 1.

To a solution of porphyrin 2 in o-dichlorobenzene was added a suspension of material 1 in o-dichlorobenzene and the reaction mixture was allowed to stir at 140 ° C for 24h.

After this time the reaction mixture was washed successively with chloroform (about 5 times) until the characteristic porphyrin soret band in the wash solvent was no longer observed.

3.2. Compounds 5, 7 and 10 were obtained by a procedure similar to that described for compound 3, with two exceptions: in the case of compounds 7 and 10, the reaction occurred in DMSO and washes of the obtained compounds were performed with:

a) chloroform / methanol (97: 3) for compound 5;

b) water for compound 7;

c) petroleum ether / dichloromethane (7: 3) for compound 10.

4. Measurement of the amount of immobilized porphyrin:

The amount of immobilized porphyrin was calculated by UV-Vis spectrophotometry.

The solvents taken from the washing of each of the reactions set out in 3.1. were diluted to a volume of 100 mL. The absorbance of each of the solutions prepared by UV-vis spectrophotometry was measured. From the Beer-Lambert law it was possible to calculate the porphyrin concentration in each solution, and thus to determine the corresponding amount of unreacted porphyrin. Since the amount of porphyrin initially added to each reaction was exactly known, the total immobilized porphyrin could be calculated by difference.

6. Biological Crops

6.1. Collectible bacterial strains

Two bacterial strains were used, one Gramnegative and one Gram-positive, respectively Escherichia coli ATCC 13706 (USA) and Enterococcus faecalis ATCC 51299 (USA).

Both strains were maintained in Triptic Soy Agar (TSA) culture medium at 4 ° C. Prior to each assay the strains were subcultured to Triptic Soy Broth (TSB) liquid medium and rechecked by 2 further subcultures to TSB medium.

6.2. Collection and treatment of wastewater samples Samples used in the tests were collected at a wastewater treatment plant (the hawaii WWTP) after secondary treatment.

Samples were collected on the days of the experiments in a sterile flask and kept in the laboratory at room temperature until analysis.

Immediately prior to analysis, the samples were filtered through a 40pm porosity nylon mesh to remove larger debris.

6.3. Production and quantification of phage stock Somatic coliphage stock was produced in Escherichia coli bacteria (ATCC 137003) from a wastewater sample subjected to secondary treatment by the Wastewater Treatment Plant (WWTP).

The sample was filtered through a 0.2 µm porosity polycarbonate membrane to remove bacteria present in the wastewater. One milliliter of the filtered sample was seeded by the double layer agar technique using as host Escherichia coli (ATCC 137003). After incubation at 37 ° C for 18 hours a plaque was selected and inoculated into 50 ml of an exponentially growing Escherichia coli culture (ATCC 137003). After 4 hours incubation at 37 ° C, the suspension was centrifuged to remove bacteria debris. The supernatant after addition of a drop of chloroform was kept at 4 ° C until use.

The phage suspension (10 ⁹ particles per milliliter) thus obtained was kept at 4 ° C.

7. Microcosm establishment

7.1. Establishment of biological samples

(a) Specimens were collected from Escherichia coli and Enterococcus faecalis collector cultures: 20 mL in a 600 mL beaker, 0.36 cm high, 10 ⁴ CFU mL ^-1 containing the photosensitizers;

(b) Wastewater samples from wastewater treatment plants were established: 20 ml in a 600 ml beaker, 0.36 cm high, containing the photosensitizers;

c) Somatic coliphage samples were established: 20 mL in a 600 mL beaker, 0.36 cm high, 10 ⁴ CFU mL ^-1 containing the photosensitizers.

7.2. Concentration in photosensitizers

Samples containing photosensitizer concentrations were established as a function of the test compounds:

(a) free porphyrin precursor 6: 0,5, 1 and 5 μΜ;

(b) derivatives 7, 8 and 11: 20 or 200 μΜ.

8. Photoinactivation Tests

Photoinactivation assays have evaluated the efficacy of some of the synthesized immobilized precursors and derivatives. For this purpose, the different microcosms containing different samples and different photosensitizers were irradiated at various concentrations.

8.1. Light irradiation

The light tests were performed under a luminaire consisting of a series of 13 lamps (380-700 nm) arranged in parallel, with a total power of 9 mW.crn ² at the distance of the lamps to which the experiments were performed.

The various biological samples containing the photosensitive compounds set forth above were irradiated with white light at 9 mW.crn ² for 60, 90, 180 and / or 270 minutes while stirring (100 rpm) at 25 °. W.

8.2. Test Controls

Were made 3 controls: dark concrete with photosensitizer (control 1), dark control without photosensitizer (control 2 ( ) and concrete without photosensitizer but exposed to same procedure in irradiation (control 3).

The assays were repeated at least twice, the results being the average of the values obtained in the various assays.

The effects of neutral material 1 and cationic material 1a on bacterial samples subjected to the same irradiation and dark conditions as with photosensitizers were also studied.

9. Measurement of results

9.1. Bacterial content of collection strains Bacterial content (contained in the different microcosms of assembled collection strains as described above) was evaluated by counting colonies after sowing by incorporation into TSA medium. From each sample were seeded by incorporating 2 1 mL replicates of the sample (at various dilutions). After 24 hours incubation in the dark at 37 ° C, colonies were counted in the most appropriate replica series.

9.2. Bacterial content in wastewater samples

CF and EF contents were determined by counting colonies on filtration membranes.

Series of 3 replicates of different sample volumes (from 1 ml of undiluted sample to 1 ml of diluted sample to 10 ~ ^{2) were established} .

These were filtered through 47mm diameter GELMAN membranes (cellulose acetate; 0.45pm porosity).

For CF the membranes were placed over agarised m-FC medium (DIFCO) and incubated in the dark at 44.5 ° C for 24 hours, and then the characteristic-looking colonies (blue-colored colonies) were counted on the replicates of more appropriate dilution.

For the PE the membranes were placed in agarized m-KF (DIFCO) medium and incubated in the dark at 37 ° C for 48 hours and then the characteristic-looking colonies (pink and red colonies) were counted in the series. replicates of the most suitable dilution.

9.3. Coliphage virus content

Quantitation of coliphage content was determined by double-layer agar technique with Escherichia coli strain (ATCC 137003).

From each sample were seeded by incorporating 2-3 replicates of 1mL of the sample (undiluted or diluted). After 18-24 hours incubation in the dark at 37 ° C phage plaques were counted in the most appropriate replica series.

EXAMPLES:

Example 1

Synthesis of silica-coated magnetic nanoparticles functionalized with propyltriethoxy silane derivatives (1)

To an aqueous solution (100 mL, pH 1; FeNH ₄ (SO ₄ ) ₂ (2.95 g; 6.12 mmol) and (NH ₄ ) ₂ Fe (SO ₄ ) ₂ (1.20 g; 3.06 mmol) At 80 ° C 25% aqueous ammonia solution (3.75 ml) was added dropwise and under magnetic stirring.

A black precipitate formed and the reaction was terminated when pH 6 was reached.

The precipitate was washed several times with deionized water and taken up in deionized water (100mL).

To the above suspension was then added, under mechanical stirring, an aqueous solution (100mL) of sodium meta-silicate pentahydrate (23.75g; 0.112mol, pH 12-13, adjusted with 37% HCl).

After addition of sodium meta silicate, the volume of the reaction mixture was adjusted to 400mL with distilled water. The resulting mixture was placed in an ultrasonic bath for 30 minutes and then progressively warmed to 80 ° C under magnetic stirring.

After cooling the mixture to room temperature, the reaction mixture was adjusted to pH (pH 6) again using 37% HCl.

The volume of the pasty-looking mixture was adjusted to 800mL with distilled H ₂ O and again subjected to ultrasound radiation for a further 30 minutes.

The mixture was then filtered through a Buckner funnel and the material washed with distilled water (2 x 200mL) and taken up in ethanol (600ml).

For the synthesis of material 1, to the above suspension was added 3-aminopropyltriethoxy silane (4mL) and the mixture was kept under magnetic stirring at room temperature and quenched after 24h. The new suspension was filtered through a Buckner funnel and the material was washed with ethanol (2 x 200mL), which was taken up in ethanol (600 mL). Material 1 has a brownish color, and by elemental analysis has a ratio of 0.149 moles of aminopropyl group / 100 grams of material.

Elemental Analysis for 1: Found (%): C, 6.117; N, 2,079; H, 2,234

Example 2

Synthesis of cationic magnetic nanoparticles (la)

To a suspension of material 1 (250 mg) in dry DMF (20mL) was added a large excess of methyl iodide (4mL).

The reaction mixture was stirred for 16 hours at 40 ° C in a round bottom flask equipped with a condenser.

After this time the reaction mixture was cooled, the product was filtered off, washed with water (3 x 50mL) and the product 1a suspended in water (25mL).

Example 3

Synthesis of 5,10,15,20-tetrakis (pentafluorophenyl) porphyrin (2)

To a mixture of glacial acetic acid (150ml) and nitrobenzene (70mL) at reflux was added pentafluorobenzaldehyde (3.6g, 29.1mmol). After dissolution of pentafluorobenzaldehyde, pyrrole (2.0mL, 28.8mmol) was added dropwise with the aid of a loading funnel. The mixture was allowed to stir at reflux (120 ° C) for one hour. After this time the reduced pressure of the glacial acetic acid and nitrobenzene was distilled to dryness. The residue was taken up in dichloromethane and chromatographed directly on a silica gel column using petroleum ether as eluent. A first fraction was obtained which was identified as 5,10,15,20-tetrakis (pentafluorophenyl) porphyrin (2). After removal of the solvent, porphyrin 2 was crystallized from dichloromethane / methanol to give purple crystals (418.0 mg, 6.6%). ^X H NMR (CDC13) δ: -2.92 (s, 2H, NH), 8.92 (s, 8H, β-Η); ¹⁹ F NMR (CDCl3) δ: -160.07 to -159.96 (m, 8F, Ar-oF), -174.77 to -174.62 (t, 4F, Ar-pF), -184.89 to 184.71 (m, 8F (Ar-mF); FAB MS * m / z: 975 (M + H) *.

Example 4

Coupling of 5,10,15,20tetrakis (pentafluorophenyl) porphyrin to material 1 (3)

To a solution of porphyrin 2 (25.0 mg, 25.7pmol) in odichlorobenzene (1mL) was added a suspension of material 1 (250 mg, 0.371 mmol of aminopropyl groups, 10 eq. Of aminopropyl groups) in o-dichlorobenzene ( 4mL).

The reaction mixture was kept under magnetic stirring at 140 ° C under nitrogen atmosphere.

The reaction was monitored by TLC and the formation of a reddish insoluble product was verified. However, after 24 hours it was found that no significant evolution of the reaction occurred, ie a considerable part of the starting porphyrin had not been consumed.

The reaction mixture was filtered and washed with 50mL volumes of chloroform until the Soret band characteristic of porphyrins in the wash solvent was no longer observed.

The determination by UV-Vis spectrophotometry of the amount of porphyrin 2 in the wash solvent allowed the difference in the amount of immobilized porphyrin to be calculated.

A suspension of compound 3 with 13.5 mg (0.036 mmol) of porphyrin 2 coupled with 250 mg of material 1 was obtained.

Example 5

Synthesis of 5-pentafluorophenyl-10,15,20-tri (4pyridyl) porphyrin (4)

To a mixture of refluxing glacial acetic acid (150mL) nitrobenzene (70 mL) was added

4-pyridinacarbaldehyde (2.5mL; 26.2mmol; 3.6 eq.) Pentafluorobenzaldehyde (0.9mL, 7.3mmol, 1 eq.

After dissolution of the aldehydes, 2mL pyrrole (28.8 mmol, 4 eq.) Was added dropwise with the aid of a loading funnel.

The mixture was refluxed for 1 hour at which time glacial acetic acid and nitrobenzene were removed to dryness under reduced pressure. The residue was taken up in 97: 3 chloroform / methanol and chromatographed directly on a silica gel column initially using 97: 3 as chloroform / methanol as eluent.

The fraction which was identified as 5- (pentafluorophenyl) -10,15,20-tri (4-pyridyl) porphyrin (4) was obtained. After removal of the solvent, porphyrin 4 was crystallized from chloroform / acetone to give purple crystals (290.5 mg, 1.4%).

^X H NMR (CDC1 ₃₎ δ: -2.91 (s, 2H, NH), 8:15 to 8:18 (m, 6H, Py-OH), 8.86 (d, 4H, β-Η, J = 5.3 Hz) 8.92 (d, 4H, β-H, J = 4.9 Hz); 9.07 (d, 6H, Py-jn-H, J = 4.7 Hz);

¹⁹ F NMR (CDCl 3) δ: -160.47 to -160.35 (m, 2F, Ar-oF), -174.35 to -175.20 (t, IF, Ar-pF), -185.20 to -186.02 (2F, m, Ar- mF);

FAB MS m / z: 708 (M + H) ⁺ .

Example 6

Coupling 5- (pentafluorophenyl) -10,15,20-tri (4pyridyl) porphyrin to material 1 (5)

To a solution of porphyrin 4 (25.7 mg, 36.3pmol) in odichlorobenzene (1mL) was added a suspension of material 1 (500 mg, 0.743mmol of aminopropyl groups, 20 eq.) In o-dichlorobenzene (9mL).

The reaction mixture was kept under magnetic stirring at 140 ° C under nitrogen atmosphere.

The reaction was monitored by TLC and after 24 hours it was found that almost all of the starting porphyrin had been consumed with the formation of a reddish insoluble product. The reaction mixture was filtered and washed with 50mL volumes of 97: 3 chloroform / methanol until no free porphyrin Soret band was observed in the wash solvent. The determination, by UV-Vis spectrophotometry,

the amount of porphyrin 4 existing in the solvent in washing allowed to calculate per difference the amount in immobilized porphyrin. The product was suspended in DMF (50mL) getting up an compound suspension 5 with 24.6 mg (0.034mmol) in porphyrin 4 coupled to 500.0 mg of material 1.

Example 7

Synthesis of 5- (pentafluorophenyl) -10,15,20tris (N-methylpyridinium-4-yl) porphyrin triiodide (6)

To a solution of porphyrin 4 (115.0 mg, 0.163 mmol) in dry DMF (10 mL) was added a large excess of methyl iodide (1 mL).

The reaction mixture was stirred for 3h at 40 ° C in a round bottom flask equipped with a condenser. After this time the mixture was cooled and the product precipitated with ethyl ether.

The precipitate was filtered through a cotton funnel and washed with ethyl ether.

residue was taken up in acetone / H ₂ 0 1: 1 and precipitated in acetone.

The methylated product was filtered off and washed with acetone and dried under vacuum to give porphyrin 6 crystals (183.1 mg,

99%).

1 H NMR (CDCl ₃ ) δ: -3.03 (s, 2H, NH), 4.71 (s, 9H, CH ₃ ) 8.99 (m, 6H, Py-oH), 9.19 (m, 6H, δ-H) 9.46-9.48 (m, 8H, 2 β-Η and 6 Py-mH);

¹⁹ F NMR (CDCl 3) δ: -163.2 to -163.1 (m, 2F, Ar-oF), -177.3 to -177.1 (t, IF, Ar-pF), -186.20 to -186.02 (m, 2F, Ar- mF); FAB MS ⁺ m / z: 752 (M + H) ⁺ .

Example 8

Coupling of 5- (pentafluorophenyl) -10,15,20tris (N-methylpyridinium-4-yl) porphyrin triiodide to material la (7)

To a solution of porphyrin 6 (41.2 mg, 36.3pmol) in DMSO (1 ml) was added a suspension of material 1 (500.0 mg, 0.743 mmol of aminopropyl groups, 20 eq.) In DMSO (9 ml).

The reaction mixture was kept under magnetic stirring at 140 ° C under nitrogen atmosphere. The reaction was monitored by TLC and after 24 hours the reaction was terminated. The reddish insoluble solid was filtered off, which was washed with water (3x50mL) until no porphyrin soret band was observed in the wash water.

The determination, by UV-Vis spectrophotometry, of the amount of porphyrin 6 present in the washing solvent, allowed to calculate, by difference, the amount of immobilized porphyrin.

solid - was resuspended in 50mL of water (for biological evaluation) or 50mL of DMF (for later cationization) to give a suspension of compound 7 with 36.6 mg (0.032mmol) of porphyrin 6 coupled to 500.0 mg of material

1.

Example 9

Preparation of cationic material (8)

a) Via cationization of material 5

To a suspension of 5 (12.3 mg porphyrin 6 coupled with 250 mg material 1) in dry DMF (20mL) was added a large excess of methyl iodide (4mL). The reaction mixture was stirred for 16 hours at 40 ° C in a flask equipped with a condenser. After this time the reaction mixture was cooled, the product was filtered and

washed up with volumes in 50mL in Water. Suspended O product resulting 8 in 25mL in Water. Obtained an suspension of compound 8th with 14, 9 mg (0.013mmol) in porphyrin 6 coupled to 250 mg of material 1 cationized.

b) Via cationization of material 7

To a suspension of 7 (18.3 mg porphyrin 6 coupled to 500 mg material 1) in dry DMF (20mL) was added a large excess of methyl iodide (2.5mL). The reaction mixture was stirred for 16 hours at 40 ° C in a round bottom flask equipped with a condenser. After this time the reaction mixture was cooled, the product was filtered off and washed with 50mL volumes of water. At the end the product was resuspended in water (25mL), obtaining a suspension of material 8 with 18.3 mg (0.016mmol) of porphyrin 6 coupled with 250 mg of catheterized material 1.

Example 10

Synthesis of 5,10,15-triphenyl-20- (pentafluorophenyl) porphyrin (9)

To a mixture of refluxing glacial acetic acid (150mL) and nitrobenzene (70mL) was added benzaldehyde (2.5mL, 24.5mmol, 3.4 eq.) And pentafluorobenzaldehyde (0.9mL, 7.3mmol, 1 eq). .).

After dissolving the two aldehydes, pyrrole (2mL, 28.8mmol, 4 eq.) Was added dropwise with the aid of a loading funnel. The mixture was refluxed for 1 hour.

After this time, distillation under reduced pressure of glacial acetic acid and nitrobenzene to dryness was initiated.

The residue was taken up in petroleum ether / dichloromethane (7: 3) and chromatographed directly on a silica gel column using petroleum ether / dichloromethane (7: 3) as eluent.

The fraction which was identified as 5,10,15-triphenyl-20- (pentafluorophenyl) porphyrin was obtained.

Porphyrin 9 was crystallized from a dichloromethane / methanol mixture to give purple crystals (307.5 mg, 1.5%).

NMR (CDC1 ₃₎ δ: -2.77 (2H, s, NH), 7.73-7.80 (9H, m, m Ar-H 6 P-3 and Ar-H), 8:19 to 8:23 (6H, m, o-Ar H), 8.76 (2H, d, βH, J = 4.7 Hz), 8.84-8.89 (4H, m, β-Η), 8.93 (2H, d, β-Η, J = 4.8 Hz).

^1H NMR F (CD01 ₃₎ δ: -160.26 ã -160.37 (m, 2F, Ar-F), -176.41 to -176.26 (t, IF, Ar-pF), -185.81 to -185.65 (m, 2F, Ar -mF);

FAB MS ⁺ m / z: 705 (M + H) ⁺

Example 11

Coupling of 5,10,15-triphenyl-20 (pentafluorophenyl) porphyrin 9 to material 1 (10)

To a solution of porphyrin 9 (26.0 mg, 36.9pmol) in DMSO (ImL) was added a suspension of material 1 (500 mg, 0.743 mmol of aminopropyl groups, 20 eq.) In DMSO (9 mL).

The reaction mixture was kept under magnetic stirring at 140 ° C under nitrogen atmosphere.

The reaction was monitored by TLC and after 24 hours it was found that almost all of the starting porphyrin had been consumed, forming a reddish insoluble product.

The reaction mixture was filtered and washed with 50mL volumes of 7: 3 petroleum ether / dichloromethane until no Soret band was observed in the wash solvent.

The product was resuspended in water (50mL) and the amount of unreacted porphyrin was calculated by UV-Vis.

A suspension of compound 10 with 20.0 mg (0.028 mmol) of porphyrin 9 coupled with 500.0 mg of material was obtained.

1.

Example 12

Cationization of material 10 (11)

To a suspension of 10 (10.0 mg (0.014 mmol) of porphyrin 9 coupled with 250 mg of material 1) in dry DMF (20 mL) was added a large excess of methyl iodide (2.5 mL).

The reaction mixture was stirred for 16 hours at 40 ° C in a flask equipped with a condenser.

After this time the reaction mixture was cooled, the product was filtered and washed with 50mL volumes of water. Product 11 was suspended in water (25mL). A suspension of compound 11 with 10.0 mg (0.014 mmol) of porphyrin 9 coupled with 250.0 mg of cationized material 1 was obtained.

REFERENCES :

[1] M. Wainwright, Journal of Antimicrobial Cnemotherapy,

1998, 42, 13.

[2] D. Faust, K.-H. Funken, G. Horneck, B. Milow, J. Ortner, M. Sattegger, M. Schâfer, C. Schmitz; Solar Energy,

1999, 65, 71.

[3] M. Krouit, R. Granet, P. Branland, B. Verneuil, P. Krausz; Bioorganic & Medicinal Chemistry Letters, 2006, 16, 1651 [4] D. A. Caminos, E. N. Durantini, Bioorganic & Medicinal Chemistry, 2006, 14, 4253.

[5] J.J. Inbaraj, Μ. V. Vinodu, R. Gandhidasan, R. Murugesan, M. Padmanabhan, Journal of Applied Polymer Science, 2003, 89, 3925.

[6] R. Bonnett, Μ. A. Krysteva, I. G. Lalov, S. V. Artarsky, Water Research, 2006, 40, 1269.

[7] M. R. Hamblin, D. A. O'Donnell, N. Murthy, K. Rajagopalan, N. Michaud,,. E. Sherwood, T. Hasan, Journal of Antimicrobial Chemotherapy, 2002, 49, 941.

[8] J. P. C. Tomé, M. G. P. M. S. Neves, A. C. Tomé, J. A.

S. Knight, M. Soncin, M. Magaraggia, S. Ferro, G. Jori, Journal of Medicinal. Chemistry, 2004, 47, 6649.

[9] TP Wijesekera, D. Dolphin, RA Sheldon, (Ed.): Metalloporphyrins in Catalytic Oxidation, New York: Mareei Dekker [10] G. Bertolini, B. Salvato, M. Dall'Acqua, M. Vazzoler, G Jori, Photochemistry and Photobiology, 1984, 39, 811.

[11] Z. Malik, J. Hanania, Y. Nitzan, Journal of

Photochemistry and Photobiology.B: Biology, 1990, 5, 281.

[12] Y. Nitzan, B. Shainberg, Z. Malik, Current

Microbiology, 1987, 15, 251.

[13] Y. Nitzan, M. Gutterman, Z. Malik, B. Ehrenberg, Pnozochemistry an d Photobiology, 1992, 55, 89. [14] Z. Malik, H. Ladan and Y. Nitzan, Journal of

Photochemistry and. Photobiology B: Biology, 1992, 14262. [151 H. Nikaido, M. Vaara, Microbiology Review., 1985, 49, 1.

[16] A. Minnock, D.I. Vernon, J. Schofield, J. Griffiths, J.H. Parish, S.B. Brown, Journal of Photochemistry and Photobiology B: Biology, 1996, 32, 159.

[17] M. Merchat, J. D. Spikes, G. Bertoloni, G. Jori, Journal of Photochemistry and. Photobiology B: Biology, 1996, 35, 149.

[18] A. Halbreich, J. Roger, J.N. Pens, D. Geldwert, M. F. Da Silva, M. Roudier, J. C. Bacri, Biochimie, 1998, 80, 379.

[19] X. Liu, Z. Ma, J. Xing, H. Liu, Journal of Magnetism and Magnetic Materials, 2004, 270, 1.

[20] M. D. Butterworth, S.A. Bell, S.P. Armes, A.W. Simpson, Journal of Colloid and Interface Science, 1996, 183, 91.

[21] S. Santra, R. Tapec, N. Theodoropoulou, J. Dobson, Langmuir, 2001, 17, 2900.

[22] K. Tao, H. Dou, K. Sun, Colloids and Surfaces A: Physicochem. Eng. Aspects, 2006, 290, 70.

[23] X. Liu, Z. Ma, J. Xing, H. Liu, Journal of Magnetism and Magnetic Materials, 2004, 270, 1.

[24] L. Shen, P. E. Laibinis, T. A. Hatton, Langmuir, 1999, 15, 447.

[25] PC Hiemenz, Principles of Colloid and Surface Chemistry, 1986, 2nd edition; Mareei Dekker: New York [26] Y. Zhang, N. Kohler, M. Zhang, Biomaterials, 2002, 23, 1553.

[27] Y. Lu, Y. Yin, Β. T. Mayers, Y. Xia, Nano Letters, 2002, 2, 183-186

28j. Santra, R. Tapec, N. Theodoropoulou, J. Doason, Langnwir, 2001, 27, 2900-2906 [29] J. I. Taylor, C. D. Hurst, M. J. Davies, N.

Sacnsinger, I. J. Bruce, J. Chromatogr. A, 2000, 890, 159.

[30] Z.-Y. Ma, X.-Q. Liu, Y.-P. Guan, H.-Z. Liu, Colloids and Surfaces A: Physicochem. Eng. Aspects, 2006, 275, 87.

[31] N. Kohler, C. Sun, J. Wang, M. Zhang, Langmuir, 2005, 21, 8858.

[32] K.M. Kadish, K.M. Smith, The Porphyrin Handbook Synthesis and Organic Chemistry, R. Guilard (Ed), Academic Press, New York, 2000, 1.

Claims (2)

1. NANOMAGNETE-PORPHYRIN HYBRIDS characterized wherein Ar, Sr and Ar'p and fenii.o, halafshilo [where rr 'means the presence of one to five halogen atoms], hydroxyphenyl, hydroxycarbonylphenyl, pyridyl to Iqu llcx loa rbon ilfen 11o, alkyl 11thiyl, alkyl 11thylphenyl, or alkyl.jpg.gicsdinso where telly means. the presence of a linear or branched aliphatic carbon bitch of varying dimensions; FEG-glridinium (where ^V PEG ^{, Z} means the presence of a variable size polyethylene chain); Ar ¹ is halophenyl or alkyl pididinium; X is aminu, oxygen, carbamate or methylene; R may be straight or branched aliphatic chain or. polyethylene of varying dimensions; 2 are functional silica coated magnetic nanoparticles bonded with amino groups at 1 qui 1 o, ha 1 o to 1 qu 1.1 o, hydroxyx 1 qu 11 o, good hydroxide 1 al qu 11 o, polyethylene 1 ico 1 and suppose i.et r seneqi r cos * 2. NANOMAGNETE PORPHYRIN HYDRIES MATERIALS according to claim 1, characterized in that they are materials preferably composed of silicon-coated magnetic particles functionalized with aminopropyl groups, some of which are covalently bonded to the following porphyrinic radicals: 5,10; 15, N-tetrakis (4-tetrafluorophenyl); phexyrinyl (3); 5- (4-tetrafluorophenyl) -10,15,20-tris (4pyridyl) porphyrinyl 5 :; 5H-tetratlucrafenylene) -10,15,20-trismethylthioyl) porphyrin (7); 5 - (4 -1 etraf 1 uorofe η 1 1 s η o) ~ 10,15 _f 2 0 ~ trife η i 1 porf 1 x 1 ni 1 o (10). 3. HYBRID MATERIALS according to claims 1 and 2 and characterized in that they are preferably constituted by magnetic nanoparticles coated with fumed silica with groups no. 1 to 11 or 11. alkyl '' '' means the presence of a linearly modified aliphatic chain of varying sizes and alkyloxy the presence of a polyethylene glycol chain of varying sizes). 4. NAKQMAILNETE-BODFIRIHA HYBRID MATERIALS, according to claim 3, characterized in that they are preferably nanoparticulated magnetic nanoparticulate materials with trimethylamide groups, i.e. some of which are attached to the following and linked together. porphyrinic radicals: 5,10,15,22-tetrakis (t-tetraflucrophenylane) pyrrinyl; 5- (4'-tetrafluorophenyl) -10,15,20-tris (4-1 / methypyridinium) porphyrinyl (8); 5 - ('4 -1 e t r a i 1 u o r o f e η 11 sη) -10., 15, · 2 0 - t r 1 f s n 11 p o r i 1 r i n i .1. o [11], Microbial agents characterized in that they are prepared with the nanomagnars-porphyrin hybrid materials of claim 1. 6. Microbial agents according to. The foregoing claim further comprises by including primarily materials consisting of the compounds of claims 2 and 5. 7. PREPARATION PROCESS for materials 3, 5, 7 and 10 described in. claim 2, characterized in that it comprises the reaction of deriratliated silica-coated magnetic nanoparticles with aminopropyl groups (methyll.l1) with 5,10,15,25-tetrakis (pentafluorophenyl) porphyrin (porphyrin 2), 5- (peniafluorophenyl) -10,15,20-t-1,4-pyridyl) porphycin (porphyrin 4), 5-pitafluorufenyl triiodide (-10,15,20thi.s (F-methylp .ii.i.diniu-R-yl) porphyrin. (porphyrin 6) and 5,10,15 -tri.fsril-20- (pentaflu.rof.enyl) porphyrin (porphyrin 9}, respectively. Preparation process of materials 3a, 8 and 11, described in claim 4, characterized in that it respectfully comprises the reaction of the nanomaguete-porphyrin hybrid materials 3, 5 and 7 (for material 0) and 10 described in nos. claim 2, with. a methylating agent. 9. USE OF NANOMAGNETE-PORPHYRIN HYDRAULATED MATERIALS, described in Claims 1 to 4, and is blended by application in water disinfection. 10. UTILITY 000 NANOMASNETE RITRICAL MATERIALS PORPHIRIN, ··· $ characterized by its application in inactivation of pathogenic microorganisms, namely bacterial cells (Gram-positive and Gram-negative}, Baorerian endospores and viruses present in water. Use of non-NOMAGNETE-PORPHYRIN HYDROCANE HOLIDAYS of formula I according to claim 1, characterized by the preparation of antimicrobial agents for tertiary treatment in ETAP / s, wastewater, swimming pools and water filters. U. USE OF NANOMAGNETEPORPHYRIN HYBRID MATERIALS IN ACCORDANCE WITH. Claim 11, characterized in that the compounds of general formula I are preferably materials consisting of the compounds of claims 2el. Use of the NANOHYAGNETE-PORPHRINE HYBRID MATERIALS according to claims 11 to 12, characterized in that they are used in filters, columns, films and membranes, preferably in the tertiary treatment of wastewater treatment plants, drinking water, swimming pools and water filters.