ITMI20111532A1 - CALIXARENIC DERIVATIVES FUNCTIONALIZED FOR CELL TRANSFORMATION - Google Patents

Description

Description of the patent for industrial invention entitled:

â € œCALIXARENE DERIVATIVES FUNCTIONALIZED FOR CELL TRANSFECTIONâ €

Technical field of invention

The present invention relates to calixarene derivatives functionalized at the upper edge (â € œupper rimâ €) or lower (â € œlower rimâ €), by means of a suitable spacer, with units of arginine or guanidinated amino acids.

The compounds of the invention are useful as non-viral vectors for cell transfection, thanks to their ability to interact non-covalently with nucleic acids.

State of the art

Gene therapy potentially represents an extraordinarily powerful medical technique for the treatment of serious diseases, of genetic but also non-genetic origin, such as tumors, neuropathies, cardiovascular diseases, hereditary diseases, etc.

Gene therapy is based on the â € œdeliveryâ € transfer of genes, a complex process consisting of several steps, some of which are not yet fully understood.

For the transfer of the gene it is necessary to have suitable vectors, able to transport the nucleic acids into the cells by crossing the membrane and to release the therapeutic material into the cytoplasm so that it can reach the appropriate compartments where it performs its function.

The most efficient vectors are viruses, which by their nature carry out transfection processes with which they infect cells, but their use is not entirely safe as it is accompanied by a series of related drawbacks, in particular high immunogenicity, toxicity and potential. mutagenic.

Furthermore, their preparation, demanding and expensive, also presents difficulties in obtaining large quantities of carriers.

For these reasons, for several years, many research groups have been dedicated to developing non-viral vectors, based on Felgner's pioneering work of the 1980s (P.L. Felgner et al., Proc. Natl. Acd. Sci. USA 1987, 84, 7413-7417). Felgner demonstrated for the first time that cationic lipids can be used to transfer DNA into mammalian cells.

Currently known and available non-viral vectors are not actually so efficient that they can be considered a definitive alternative to viruses, even if they have, in general, the advantage of not being immunogenic and often weakly toxic. Furthermore, they can in principle be structurally modified without limits, in order to optimize their properties as gene transport systems. These negative and positive aspects that characterize them, considered together, justify the still considerable interest both in the academic world and in the industry for this research sector and for the development of new non-viral vectors.

Non-viral vectors generally consist of cationic lipids or cationic polymers and, more recently, dendrimers and nanoparticles have also been used. Typically, especially cationic lipids, they are used in transfection protocols suitably mixed with neutral lipids, called "helper lipids", such as dioleoyl-glycerophosphatidylethanolamine (DOPE) and dioleoyl-glycerophosphatidylcholine (DOPC). The first of these is one of the neutral lipids currently most used in gene therapy applications. These neutral lipids help the transfection process by improving efficiency and limiting cytotoxicity, but their use in transfection formulations is definitely a complication. It would therefore be useful to be able to avoid its use in commercial products to simplify the protocols.

Calix [n] arenes are cyclic oligomeric compounds, characterized by the presence of a cavity, which can be easily functionalized both at the upper (â € œupper rimâ €) and lower (â € œlower rimâ €) edge of this cavity.

These compounds are described for example in: a) Calixarenes 2001; Asfari, Z., Böhmer, V., Harrowfield, J., Vicens, J., Eds .; Kluwer Academic: Dordrecht, 2001; b) Calixarenes in Action; Mandolins, L .; Ungaro, R. eds; Imperial College: London, 2000; c) C. D. Gutsche, Calixarenes. An Introduction, and J. F. Stoddart, The Royal Society of Chemistry, Cambridge, 2008, d) Böhmer, V. Angew. Chem., Int. Ed. Engl. 1995, 34, 713â € “745; e) Pochini, A .; Ungaro, R. In Comprehensive Supramolecular Chemistry. Vögtle, F., Ed .; Elsevier Science: Oxford, 1996; Vol. 2, p 103; f) Ikeda, A .; Shinkai, S. Chem. Rev. 1997, 97, 1713â € “1734; g) Baldini, L .; Casnati, A .; Sansone, F .; Ungaro, R. Chem. Soc. Rev. 2007, 36, 254-266; h) Samson, F .; Baldini, L .; Casnati, A .; Ungaro, R. New J. Chem. 2010, 34, 2715-2728.

Guanidinated derivatives based on calix [4] arenica bearing adamantine groups at the â € œlower rimâ € and potentially usable in a different field such as that of nanolithography are described in Angew. Chem. Int. Ed. 2004, 43, 369-373 and in J. Am. Chem. Soc. 2004, 126, 6627-6636. These compounds are able to interact with self-assembled monolayers of beta-cyclodextrin following the inclusion of the adamantyl group in the lipophilic cavity of the oligosaccharide macrocycle.

Compounds endowed with translocation properties across cell membranes and epithelial tissues and consisting of a central structure with a plurality of side chains containing guanidine groups are described in WO 2007/038169. Calix [n] arenic structures (n = 4 or 6) are reported as examples of the aforementioned central structure. In these structures, the guanidine group is covalently linked, in the â € œlower rimâ €, to an organic fragment, which in turn is linked through a spacer to the phenolic oxygen of the calix [n] arene. However, the preparation of any compound with a calix [n] arenic structure substituted by guanidine groups is not described. Furthermore, the therapeutic molecules that are indicated are intended to be covalently linked to the central structure.

Peptidomimetics based on the calix [4] arenic structure and substituted for phenolic oxygen with guanidino-alkyl residues are described in US2008 / 0300164 A1. The compounds are described as having various biological activities, such as, for example, bactericidal, antiangiogenic and / or antitumor activity but without any reference to transport functions for nucleic acids.

Calix [n] arenes (n = 4, 6 or 8) bearing guanidine groups directly linked to the aromatic nucleus (at the "upper rimâ €) of the calixarene structure and alkyl chains of different lengths (methyl, propyl, octyl) on the phenolic oxygens at â € œlower rimâ € are described in J. Am. Chem. Soc. (2006), 128, 14528-14536 and in Tetrahedron, (2004), 60, 11613-11618. Some of the compounds are capable of condensing plasmid DNA and linear and in some cases they promote cell transfection with an efficacy depending on the size of the macrocycle, on the conformation (cone, 1,3-alternating or mobile) and on lipophilicity. In general, the transfection efficiency of these compounds is quite low Compounds are also endowed with high cytotoxicity especially at the concentrations required to achieve transfection.

Therefore, there is still a need to identify new compounds that can be used as non-viral vectors for cell transfection.

List of figures

Figure 1 shows the transfection efficacy of an â € œupper rimâ € argininocalixarene against Rhabdomiosarcoma (RD-4) cells in the presence and absence of the auxiliary lipid DOPE, compared to Lipofectamine LTX <â „¢> and DOPE employed alone.

Figure 2 shows the transfection efficacy against Rhabdomiosarcoma (RD-4) cells (in terms of percentage of cells transfected with the pEGFP-P1 plasmid) of an â € œupper rimâ € argininocalixarene in the presence and absence of auxiliary lipid DOPE, compared to calixarenes bearing simple guanidinium groups, to non-macrocyclic analogues and to Lipofectamine LTX <â „¢> and DOPE used alone.

Figure 3 shows the transfection efficacy against Rhabdomiosarcoma (RD-4) cells (in terms of Relative Units of Luciferase) of an â € œupper remâ € argininocalixarene in the presence and absence of the auxiliary DOPE lipid, compared to calixarenes bearing simple guanidinium groups, to non-macrocyclic analogs, and compared to Lipofectamine LTX <â „¢> and DOPE used alone.

Figure 4 shows cytotoxicity measures related to an “upper rim” argininocalixarene using Rhabdomiosarcoma (RD-4) cells.

Figure 5 shows the efficacy of transfection against VERO cells of an â € œupper rimâ € argininocalixarene in the presence and absence of the auxiliary lipid DOPE, compared to Lipofectamine LTX <â „¢> used alone.

Figure 6 shows the efficacy of transfection against N2a cells of an â € œupper rimâ € argininocalixarene in the presence and absence of the auxiliary lipid DOPE, compared to Lipofectamine LTX <â „¢> used alone.

Figure 7 shows the efficacy of transfection against EADSC cells of an â € œupper rimâ € argininocalixarene in the presence and absence of the auxiliary lipid DOPE, compared to Lipofectamine LTX <â „¢> used alone.

Detailed description of the invention

It has now been surprisingly found that calix [n] arenic derivatives functionalized at the upper edge (â € œupper rimâ €) or lower (â € œlower rimâ €), through a suitable variable spacer, with units of arginine or guanidinated amino acids, are endowed with advantageous characteristics of cell transfection and low cytotoxicity with respect to the calix [n] arenic derivatives previously described and with respect to commercially available formulations such as Lipofectamine LTX <TM>.

The present invention relates to compounds of general formula (I):

FGFG

m

S.

FG FG

OR

S OR RO S

OR

FG FG

m m

n

S.

FGFG

m (I)

in which:

n is an integer from 1 to 5, preferably n is 1, 3 or 5;

m is 0 or 1;

each R is independently an alkyl chain containing from 1 to 12 carbon atoms optionally functionalized with hydroxyl groups, C1-C4 alkoxy groups, amino groups or aryl groups; a polyoxyethylene chain (CH2-CH2-O) pORâ € ™ in which p is an integer from 1 to 6 and Râ € ™ is hydrogen or C1-C2 alkyl; or each R is independently an alkyl chain containing from 1 to 12 carbon atoms optionally functionalized with an amidine group, or with a guanidine group or a bicyclic guanidine group having formula:

H Rc

N N

Ra Rd N

H N

NOT

Rb N

or HH

in which Ra, Rb, Rc and Rd are independently selected from hydrogen, C1-C4 alkyl, or Rc and Rd taken together with the nitrogen atom to which they are bound can form a pyrrolidine, piperidine, morpholine or piperazine ring.

According to a preferred aspect, R is an alkyl chain containing from 1 to 8 carbon atoms, even more preferably a hexyl or octyl chain or a 2-ethoxyethoxy or 2-methoxyethoxy chain.

When m = 0, S is a spacer chosen from the group consisting of -C (= O) NH (CH2) r-, -NHC (= O) (CH2) r-, -CH2NH (CH2) r-, -CH2O (CH2) r-, -CH2S (CH2) r-, -CH2S-S (CH2) r-, -NHC (= O) NH (CH2) r-, -NHC (= S) NH (CH2) r-, where r is zero or an integer from 1 to 3, -NHC (= O) (CH2) q- (1,2,3-triazolyl) - (CH2) râ € ™ - where q is zero or a integer from 1 to 4 and râ € ™ is an integer from 1 to 3, - (CH2) s- (1,2,3-triazolyl) - (CH2) qC (= O) NH (CH2) r- in which s is zero or 1, q and r are as defined above, -C (= O) OCH2-, -CH = NOCH2-, where 1,2,3-triazolyl is a substituted 1,2,3-triazole in positions 1 and 4 or 1 and 5 by the groups indicated in the above formulas of S; the initial atom of S starting from the left is understood to be linked to FG;

S is a spacer chosen from the group consisting of (-NHC (= O) CH2) 2NC (= O) (CH2) r-, (-NHC (= O) CH2) 2CHNHC (= O) (CH2) r-, (-C (= O) NHCH2CH2) 2NC (= O) (CH2) r- o (-C (= O) NH) 2CHC (= O) NH (CH2) r, where r is as defined above, when m = 1; the initial atom of S starting from the left is understood to be linked to FG.

Preferably S is -C (= O) NH (CH2) r- in which r is as defined above; the initial atom of S starting from the left is understood to be linked to FG.

FG is a group of formula:

CG

CH2

q

R1

in which:

q is zero or an integer from 1 to 4;

CG is an amidine group, or a guanidine group or a bicyclic guanidine group having the formula:

H Rc

N N

Ra Rd N

H N

N O N N

Rb

or HH

wherein Ra, Rb, Rc and Rd are as defined above;

R1 is hydrogen, an amino group, an ammonium group, an acylated amino group, a guanidine group as defined above when S is bonded to FG by means of a carbon atom; a carboxylic group or a carboxylic derivative such as for example a carboxylate, carboxyamide, N- (C1-C4-alkyl) carboxyamide group or N, N-bis- (C1-C4-alkyl) carboxyamide, C1-C4 alkoxycarbonyl, when S Ã ̈ bonded to FG by a nitrogen atom.

Each guanidine group as defined above and each amidine group, present in R, R1 and in the CG group, is protonated and each amino group, present in R, R1 and in the CG group, can be protonated and in this case their positive charge is It is neutralized by a counteranion which can be chosen from halide, sulphate, nitrate, carbonate, phosphate, trifluoroacetate, acetate.

The calixarene structure of formula (I) can be in different conformations, cone, 1,3-alternating, 1,2-alternating, partial or mobile cone, and can be constituted by 4 (n = 1), 5 (n = 2), 6 (n = 3), 7 (n = 4) or 8 (n = 5) phenolic rings.

When R1 is different from hydrogen the FG group is chiral, therefore the present invention also includes the single enantiomers and the diastereoisomers of the compounds of formula (I).

According to a preferred aspect of the invention, in the compounds of formula (I) FG is a group of formula:

NH2

H N NH2

CH2

q

R1

in cu R1 and q are as defined above.

According to a further preferred aspect of the invention, in the compounds of formula (I), n à 1 and m à 0, S is the group -C (= O) NH- and FG is a group of formula:

NH

H N NH3Cl

CH2

q '

Cl H3N

where qâ € ™ is 1, 2 or 3.

Preferred compounds of general formula (I) are, for example:

5,11,17,23-Tetrakis (L-argininoylamino) -25,26,27,28-tetra (nesyloxy) calix [4] arene octa-hydrochloride, in which, on the basis of the general formula I, n = 1, m = 0, R is n-hexile, S Ã ̈ C (= O) NH, FG is such that R1 is an ammonium group, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23,29,35-Esakis (L-argininoylamino) -25,26,27,28,29,30-exa (nesyloxy) calix [6] arene dodeca-hydrochloride, in which, on the basis of general formula I, n = 3, m = 0, R is n-slender, S is C (= O) NH, FG is such that R1 is an ammonium group, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23,29,35,41,47-Octakis (L-argininoylamino) -49,50,51,52,53,54,55,56-octa (n-hexyloxy) calix [6] arene exadeca-hydrochloride, in which, on the basis of the general formula I, n = 5, m = 0, R is n-slender, S Ã ̈ C (= O) NH, FG is such that R1 is a group ammonium, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23-Tetrakis (4-guanidinobutanoylamino) -25,26,27,28-tetra (nesyloxy) calix [4] arene tetra-hydrochloride, in which, on the basis of the general formula I, n = 1, m = 0, R is n-hexile, S Ã ̈ C (= O) NH, FG is such that R1 is hydrogen, q = 2 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23-Tetrakis (4-guanidinobutanoylamino) -25,26,27,28-tetra (noctyloxy) calix [4] arene tetra-hydrochloride, in which, on the basis of the general formula I, n = 1, m = 0, R is n-octyl, S Ã ̈ C (= O) NH, FG is such that R1 is hydrogen, q = 2 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23-Tetrakis (L-argininoylamino) -25,26,27,28-tetra (npropyloxy) calix [4] arene octa-hydrochloride, in which, on the basis of the general formula I, n = 1, m = 0, R is n-propyl, S Ã ̈ C (= O) NH, FG is such that R1 is an ammonium group, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23-Tetrakis (L-argininoylamino) -25,26,27,28-tetrakis [(2-ethoxy) ethoxy] calix [4] arene octa-hydrochloride, in which, based on the general formula I , n = 1, m = 0, R is 2-ethoxyethyl, S is -C (= O) NH-, FG is such that R1 is an ammonium group, q = 3 and CG is the group guanidinium as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23-Tetrakis (L-argininoylaminomethyl) -25,26,27,28-tetra (nesyloxy) calix [4] arene octa-hydrochloride, in which, on the basis of the general formula I, n = 1, m = 0, R is n-slender, S Ã ̈ -C (= O) NHCH2-, FG is such that R1 is an ammonium group, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23-Tetrakis (L-argininoylaminomethyl) -25,26,27,28-tetra (noctyloxy) calix [4] arene octa-hydrochloride, in which, on the basis of the general formula I, n = 1, m = 0, R is n-octyl, S is ̈ -C (= O) NHCH2-, FG is such that R1 is an ammonium group, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23-Tetrakis [(2-amino-5-guanidinopentyl) aminomethyl] -25,26,27,28-tetra (n-hexyloxy) calix [4] arene dodeca-hydrochloride, in which, on the basis of the general formula I, n = 1, m = 0, R is n-slender, S is â € “CH2NHCH2-, FG is such that R1 is an ammonium group, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23-Tetrakis [(2-amino-5-guanidinopentyl) aminomethyl] -25,26,27,28-tetra (n-octyloxy) calix [4] arene dodeca-hydrochloride, in which, on the basis of the general formula I, n = 1, m = 0, R is n-octyl, S is â € “CH2NHCH2-, FG is such that R1 is an ammonium group, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23-Tetrakis [di (L-argininoylamino) acetylamino] -25,26,27,28-tetra (n-hexyloxy) calix [4] arene exadeca-hydrochloride, in which, based on the general formula I, n = 1, m = 1, R is n-slender, S Ã ̈ (â € “C (= O) NH) 2CHC (= O) NH-, FG is such that R1 is a group ammonium, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23-Tetrakis [di (L-argininoylamino) acetylamino] -25,26,27,28-tetra (n-octyloxy) calix [4] arene exadeca-hydrochloride, in which, based on the general formula I, n = 1, m = 1, R is n-octyl, S Ã ̈ (â € “C (= O) NH) 2CHC (= O) NH-, FG is such that R1 is a group ammonium, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23-Tetrakis [N, N-di (2-L-argininoylaminoethyl) aminocarbonyl] -25,26,27,28-tetra (nesyloxy) calix [4] arene exadeca-hydrochloride, in which, on basis of the general formula I, n = 1, m = 1, R Ã ̈ n-slender, S Ã ̈ (â € “C (= O) NHCH2CH2) 2NC (= O) -, FG is such that R1 Ã ̈ an ammonium group, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

5,11,17,23-Tetrakis [N, N-di (2-L-argininoylaminoethyl) aminocarbonyl] -25,26,27,28-tetra (noctyloxy) calix [4] arene exadeca-hydrochloride, in which, on basis of the general formula I, n = 1, m = 1, R Ã ̈ n-octyl, S Ã ̈ (â € “C (= O) NHCH2CH2) 2NC (= O) -, FG is such that R1 Ã ̈ an ammonium group, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

The present invention also relates to compounds of general formula (II):

R1R1

S1 S1

m

CF

R1 R1

S1 S1

OR m

CF OR RO CF

OR

S1 S1

R1

R1

m CF no

S1S1

m

R1R1

(II)

in which

n is an integer from 1 to 5, preferably n is 1, 3 or 5;

m is 0 or 1;

each R is independently an alkyl chain containing from 1 to 12 carbon atoms optionally functionalized with hydroxyl groups, C1-C4 alkoxy groups, amino groups or aryl groups; a polyoxyethylene chain (CH2-CH2-O) pORâ € ™ in which p is an integer from 1 to 6 and Râ € ™ is hydrogen or C1-C2 alkyl; or each R is independently an alkyl chain containing from 1 to 12 carbon atoms optionally functionalized with an amidine group, or with a guanidine group or a bicyclic guanidine group having formula:

H Rc

NN N

Ra Rd H

No.

N O N N

Rb or HH

in which Ra, Rb, Rc and Rd are independently selected from hydrogen, C1-C4 alkyl, or Rc and Rd taken together with the nitrogen atom to which they are bound can form a pyrrolidine, piperidine, morpholine or piperazine ring.

According to a preferred aspect of the invention, R is an alkyl chain containing from 1 to 8 carbon atoms, even more preferably a hexyl or octyl chain or a 2-ethoxyethoxy or 2-methoxyethoxy chain.

CF is a spacer chosen from the group consisting of -C (= O) (CH2) r-, -NH (CH2) r-, where r is zero or an integer from 1 to 3, for m = 0; the initial CF atom starting from the left is understood to be linked to S1;

CF is a spacer chosen from the group consisting of (-C (= O) CH2) 2NC (= O) (CH2) s-, (-C (= O) CH2) 2CHNHC (= O) (CH2) s-, (-NHCH2CH2) 2NC (= O) (CH2) s- o (- (NH) 2CHC (= O) NH (CH2) r, where s is zero or 1 and r is zero or an integer from 1 to 3, for m = 1, the initial CF atom starting from the left is understood to be linked to S1;

S1 is selected from the group consisting of a divalent radical of an oligopeptide consisting of (L) and / or (D) -amino acids and / or non-chiral amino acids, containing at least one arginine or a guanidinate amino acid in which the guanidine group has formula:

H Rc

NN N

Ra Rd H

No.

N O N N

Rb or HH

wherein Ra, Rb, Rc and Rd are as defined above; and in which S1 is linked to CF and R1 through its N- and C- terminal ends, or C- and N-terminals, respectively.

When S1 is linked to R1 through the N-terminal end, R1 is hydrogen, a C1-C4-acyl group, a C1-C4-alkoxycarbonyl group or a group having the formula:

H Rb N

N N

Ra Rc O N N +

or H H

When S1 is bonded to R1 through the C-terminal end, R1 is a hydroxyl group or C1-C4-alkoxide or, taken together with the C-terminal carbonyl group of S1, R1 forms a carboxyamide group, N- ( C1-C4-alkyl) carboxyamide or N, N-bis- (C1-C4-alkyl) carboxyamide.

Each guanidine group as defined above and each amidine group, present in R, S1 and R1, is protonated and each amino group present in R, S and R1, can be protonated and in this case their positive charge is neutralized by a counteranion which can be chosen from halide, sulphate, nitrate, carbonate, phosphate, trifluoroacetate, acetate.

The calixarene structure of formula (II) can be in different conformations, cone, 1,3-alternating, 1,2-alternating, partial or mobile cone, and can be constituted by 4 (n = 1), 5 (n = 2), 6 (n = 3), 7 (n = 4) or 8 (n = 5) phenolic rings.

Preferred compounds of general formula (II) are, for example:

- 5,11,17,23-Tetrakis (L-arginyl-glycinoylamino) -25,26,27,28-tetra (nesyloxy) calix [4] arene octa-hydrochloride, in which, on the basis of the general formula II, n = 1, m = 0, R is n-hexyl, CF -NH-, S1 is L-arginylglycinoyl, R1 is hydrogen;

- 5,11,17,23-Tetrakis (L-arginyl-glycinoylamino) -25,26,27,28-tetra (n octyloxy) calix [4] arene octa-hydrochloride, in which, on the basis of the general formula II, n = 1, m = 0, R is n-octyl, CF -NH-, S1 is L-arginylglycinoyl, R1 is hydrogen;

- 5,11,17,23-Tetrakis (L-arginyl-L-argininoylamino) -25,26,27,28-tetra (n-hexyloxy) calix [4] arene dodeca-hydrochloride, in which, based on the formula general II, n = 1, m = 0, R is n-hexyl, CF -NH-, S1 is L-arginyl-L-argininoyl, R1 is hydrogen;

- 5,11,17,23-Tetrakis (L-arginyl-L-argininoylamino) -25,26,27,28-tetra (n-octyloxy) calix [4] arene dodeca-hydrochloride, in which, based on the formula general II, n = 1, m = 0, R is n-octyl, CF -NH-, S1 is L-arginyl-L-argininoyl, R1 is hydrogen.

The present invention also relates to compounds of general formula (III):

FGFG

m

S2

OR

FG

S2

R2

FG <O> R2 R2 O FG

m R2

S2

FG

m

OR

n

S2

FGFG

m (III) in which

n is an integer from 1 to 5, preferably n is 1, 3 or 5;

m is 0 or 1;

each R2 is independently hydrogen; an alkyl chain containing from 1 to 12 carbon atoms optionally functionalized with hydroxyl groups, C1-C4 alkoxy groups or with amino groups; or R2 is an aryl or aryloxy group or an aryl-C1-C4 alkyl chain. R2 is preferably a C6-C8 alkyl or benzyl or C6H5 chain.

When m = 0, S2 is a spacer chosen from -NHC (= O) CH2-, -C (= O) NH (CH2) s (CH2CH2O) pâ € ™ (CH2) t-, -CH2NH (CH2) s (CH2CH2O) pâ € ™ (CH2) t-, -NHC (= O) NH (CH2) s (CH2CH2O) pâ € ™ (CH2) t-, -NHC (= S) NH (CH2) s (CH2CH2O) pâ € ™ (CH2) t-, where s is zero or 1, pâ € ™ is zero or an integer from 1 to 6, t is an integer from 2 to 4, - (CH2) s- (1, 2,3-triazolyl) - (CH2) tâ € ™ - where s is zero or 1 and tâ € ™ is 1 or 2, -NHC (= O) (CH2) q- (1,2,3- triazolil) - (CH2) s (CH2CH2O) pâ € ™ (CH2) t- where q is zero or an integer from 1 to 4, s is zero or 1, pâ € ™ is zero or an integer from 1 a 6, t is an integer from 2 to 4, 1,2,3-triazolil is a 1,2,3-triazole substituted in positions 1 and 4 or 1 and 5 by the groups indicated in the above S2 formulas; the initial atom of S2 starting from the left is understood to be linked to FG;

S2 is a spacer chosen from (-C (= O) CH2) 2NC (= O) CH2-, (-C (= O) CH2) 2CHNHC (= O) CH2-, (-C (= O) NH) 2CHC (= O) NH (CH2) 3) - with m = 1; the initial atom of S2 starting from the left is understood to be linked to FG;

FG is a group of formula:

CG

CH2

q

R1

where q is zero or an integer from 1 to 4;

CG is an amidine group, or a guanidine group or a bicyclic guanidine group having the formula:

Rc

N N N

Ra Rd H

No.

N O N N

Rb or HH

in which Ra, Rb, Rc and Rd are independently selected from hydrogen, C1-C4 alkyl, or Rc and Rd taken together with the nitrogen atom to which they are bound, they can form a pyrrolidine, piperidine, morpholine or piperazine ring;

R1 is hydrogen, an amino group, an ammonium group, an acylated amino group, a guanidine group as defined above, a carboxylic group or a carboxylic derivative such as for example a carboxylate, carboxyamide, N- (C1-C4-alkyl) group carboxyamide, N, N-bis- (C1-C4-alkyl) carboxyamide or C1-C4 alkoxycarbonyl, provided that, when S2 is bonded to FG by a nitrogen atom, R1 is hydrogen or a carboxylic derivative as defined above .

Each guanidine group as defined above and each amidine or amino group present in R, R1 and in the CG group, can be protonated and in this case their positive charge is neutralized by a counteranion which can be chosen among halide, sulphate, nitrate, carbonate, phosphate, trifluoroacetate, acetate.

The calixarene structure of formula (III) can be in different conformations, cone, 1,3-alternating, 1,2-alternating, partial or mobile cone, and can be constituted by 4 (n = 1), 5 (n = 2), 6 (n = 3), 7 (n = 4) or 8 (n = 5) phenolic rings.

When R1 is different from hydrogen the FG group is chiral, therefore the present invention also includes the single enantiomers and the diastereosisomers of the compounds of formula (III).

Preferred compounds of general formula (III) are, for example:

- 5,11,17,23-Tetra-n-hexyl-25,26,27,28-tetrakis [3- (N-L-argininoylamino) propoxy] calix [4] arene octa-hydrochloride, in which, based on the formula general III, n = 1, m = 0, R2 is n-slender, S2 is â € “C (= O) NH (CH2) 3-, FG is such that R1 is an ammonium group, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

- 5,11,17,23-Tetra-n-hexyl-25,26,27,28-tetrakis [N, N-di (L-argininoylmethyl ester) carbonylmethyl] -3-aminopropoxicalx [4] arene octachlorhydrate, in which, based on the general formula III, n = 1, m = 1, R2 is n-slender, S2 is (â € “NHC (= O) CH2) 2N (CH2) 3-, FG is such that R1 Is the methoxycarbonyl group, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen;

- 5,11,17,23-Tetra-n-hexyl-25,26,27,28-tetrakis {N, N-di [2- (L-argininoylamino) ethyl] aminocarbonylmethoxy} calix [4] arene exadecachhydrochloride, in which, on the basis of the general formula III, n = 1, m = 1, R2 is n-slender, S2 is (â € “C (= O) NHCH2CH2) 2NC (= O) CH2-, FG is such for which R1 is the ammonium group, q = 3 and CG is the guanidinium group as defined above with Ra = Rb = Rc = Rd = hydrogen.

As shown in the examples, the compounds of the invention are efficient vectors for the introduction of nucleic acids into cells and are also characterized by low cytotoxicity.

Another important aspect is that the compounds of the invention are characterized by a high transfection efficiency and low toxicity, without being co-formulated with the lipid helper DOPE (dioleoylphosphatidylethanolamine), which is actually used in many Commercially available formulations for transfection experiments.

The presence of arginine and guanidinated amino acid units in the calix [n] arenic structure vectors determines a peculiar and drastic improvement in the quality of the vectors in terms of transfection efficiency and cytotoxicity decrease, in particular compared with simply guanidinated calixarenes described in the state of the technique.

A further object of the invention is therefore the use of a compound of formula (I), (II), or (III) as a non-viral vector for cell transfection.

Another object of the invention are compositions comprising at least one compound of formula (I) or (II) or (III) in a suitable vehicle. Examples of suitable vehicles for the formulation of compounds of the invention are water and / or short-chain water / alcohol mixtures in variable percentages depending on the lipophilicity of the compound.

Examples

Example 1 - Method of preparation

The compounds of formula (I) or (II) can be prepared starting from the corresponding p-tert-butylcalix [n] arenes by alkylation of the phenolic hydroxyl groups with chains of different length and nature using, when possible, the suitable conditions to block macrocycles in defined conformations, well known in the chemistry of phenols and calixarenes. The tert-butyl groups present on the upper rim of the aromatic rings are replaced by nitro groups by hypso-nitration, and are subsequently reduced to amino groups necessary for condensation with arginine or other amino acid units. Alternatively, the amino groups, or other functional groups suitably introduced to the â € upper rimâ € by replacing the tert-butyl groups through well-known procedures of the chemistry of calix [n] arenes and phenols, are reacted to introduce spacers that bear one or more reactive centers for coupling with one or more units of arginine or another amino acid. Arginine and other amino acids are used in protected form and are purchased as commercially available reagents or prepared according to the typical protective reactions of peptide chemistry. The deprotection reactions for obtaining the final products are carried out under the usual conditions reported for peptide chemistry. The amino acids different from arginine are guanidinated to the nitrogen atom using appropriately activated and protected guanidines, or N-protected thioureas, using methods well known in the art.

For the synthesis of the compounds of formula (III) which bear guanidine groups at the lower â € œlower rimâ €, the phenolic hydroxyl groups of the calix [n] arenes are alkylated with chains ending with one or more reactive groups necessary for the coupling with one or more units of arginine or another amino acid suitably protected with the usual protective groups used in peptide synthesis.

To the â € ™ upper rimâ € different chains are introduced following the typical procedures of calix [n] arene chemistry, based on the chemistry of phenols and, more generally, of aromatic compounds.

The introduction on the calixarene structure of the amino acid that carries or to which the charged group CG will be linked can also be performed through the formation of groups other than the amide group, such as through triazole rings or through the imino, amino, urea groups , thiourea, ether, thioether or disulfide, carrying out coupling reactions with amino acids suitably modified to the carboxyl group, which can be previously reduced to CH2OH and used as such or further transformed, for example, into CH2Br, CH2N3, or CH2NH2, or be reduced to CHO aldehyde group and reacted via reductive amination.

Example 2 - General procedure used for the synthesis of calix [n] arenic derivatives functionalized on the upper or lower edge (â € œuper or lower rimâ €) with protected arginine units

To a suitably protected arginine dimethylformamide (DMF) solution (1.5 eq. For each calixarene group to be functionalized) under stirring, hydroxybenzotriazole (HOBt) (1.7 eq. For each calixarene group to be functionalized) is added and dicyclohexylcarbodiimide (DCC) (1.5 eq. for each group of calixarene to be functionalized). After 15 minutes a solution of aminocalix [n] arene (1 eq.) In DMF is added drop by drop. The mixture is stirred at room temperature for 24 hours. Ethyl acetate is added, the dicylohexylurea (DCU) is removed by filtration through a Hirsch funnel, the organic phase is washed with a saturated aqueous solution of NaHCO3, brine (3×) and dried over Na2SO4 anhydrous. The solvent is removed under reduced pressure giving a raw material which is purified as indicated below.

Example 3 - 5,11,17,23-Tetrakis [(NÎ ± -Boc-NÏ ‰ -Tos-L argininoyl) amino] -25,26,27,28-tetra (n-hexyloxy) calix [4] arene Il reaction crude obtained according to the general procedure of Example 2 is purified by preparative TLC (eluent: CH2Cl2 / MeOH = 94/6) to obtain the pure product in the form of a white solid.

<1> H NMR (300 MHz, CD3OD) Î ́ 7.71 (d, J = 7.9 Hz, 8H, Tos), 7.23 (d, J = 7.9 Hz, 8H, Tos), 7.03 (bs, 4H, ArH), 6.72 (bs, 4H, ArH), 4.47 (d, J = 13.0 Hz, 4H, ArCH2Ar), 4.10 (bs, 4H, COCHNH), 3.91 (t, J = 6.9 Hz, 8H, ArOCH2), 3.18 (bs, 8H, CH2NH), 3.13 (d, J = 13.0 Hz, 4H, ArCH2Ar), 2.32 (s, 12H, CH3Tos), 2.00-1.88 (m, 8H, OCH2CH2), 1.80-1.25 (m, 76H, OCH2CH2CH2CH2CH2, CH2CH2CH2NH and tBu), 0.96 (t, J = 6.7 Hz, 12H, CH2CH3). <13> C NMR (75 MHz, CD3OD) Î ́ 173.1, 160.4, 159.9, 155.1, 143.8, 142.4, 136.6, 133.5, 130.6, 127.4 , 123.1, 122.2, 81.1, 76.8, 56.4, 40.1, 33.7, 32.5, 31.7, 31.2, 29.2, 27.6, 24.3, 21.7, 14.8. MS (ESI): calculated for [M 2Na] <++> m / z = 1253.6, found m / z = 1254.2.

Example 4 - 5,11,17,23-Tetrakis [(NÎ ± -Fmoc-NÏ ‰ âˆ'Mtr-L-argininoyl) amino] -25,26,27,28-tetra (n-hexyloxy) calix [4] arenas

The raw reaction obtained according to the general procedure of example 2 is purified by column chromatography (eluent: ethyl acetate / hexane = 4/1) to obtain the pure product in the form of a white solid.

Melting point: 160-162 ° C. <1> H NMR (300 MHz, CD3Cl3 / CD3OD = 19: 1) Î ́ 8.97 (bs, 4H, ArNH), 7.66 (d, J = 7.1 Hz, 8H, ArFmoc ), 7.48 (d, J = 5.4 Hz, 8H, ArFmoc), 7.30 (t, J = 7.1 Hz, 8H, ArFmoc), 7.25-7.10 (m, 8H, ArFmoc), 6.96 (bs, 4H, ArH), 6.42 (bs, 8H, ArH and ArMtr), 4.40 (d, J = 12.4 Hz, 4H, ArCH2Ar), 4.35-4.10 (m, 12H, CH2CHFmoc), 4.10-4.00 (m, 4H, COCHNH), 3.84 (bs , 8H, ArOCH2), 3.69 (s, 12H, OCH3Mtr), 3.30-3.00 (m, 12H, ArCH2Ar and CH2NH), 2.62 (s, 12H, CH3Mtr), 2.56 (s, 12H, CH3Mtr), 2.02 (s, 12H, CH3Mtr), 1.91 (bs, 8H, OCH2CH2), 1.85-1.25 (m, 40H, OCH2CH2CH2CH2CH2and CH2CH2CH2NH), 0.95-0.83 (m, 12H, CH2CH3).

<13> C NMR (75 MHz, CD3Cl3 / CD3OD = 19: 1) Î ́ 170.8, 158.4, 156.5, 156.4, 153.6, 143.7, 143.5, 141.1, 138.4, 136.4, 135.1, 133.0, 127.6, 127.0, 125.0, 124.8 , 121.5, 119.8, 111.7, 75.5, 66.9, 55.3, 55.0, 46.9, 39.8, 32.0, 31.0, 30.0, 29.6, 25.8, 25.3, 24.0, 22.7, 18.3, 14.0, 11.8. MS (ESI): calculated for [M 2Na] <++> m / z = 1613.7, found m / z = 1614.5.

Example 5 - 5,11,17,23-Tetrakis [(NÎ ± -Boc-NÏ ‰ âˆ'Pbf-L-argininoyl) amino] -25,26,27,28-tetra (n-hexyloxy) calix [4] arenas

The raw reaction obtained according to the general procedure of Example 2 is purified by preparative TLC (eluent: CH2Cl2 / MeOH = 94/6) to obtain the pure product in the form of a white solid.

Melting point: 156-158 ° C. <1> H NMR (300 MHz, CD3Cl3 / CD3OD = 19: 1) Î ́ 9.04 (bs, 4H, ArNH), 7.10-6.10 (bs, 8H, ArH and NHCNHNH) , 5.90 (bs, NHBoc), 4.33 (d, J = 13.1 Hz, 4H, ArCH2Ar), 4.06 (bs, 4H, COCHNH), 3.77 (bs, 8H, OCH2), 3.10 (bs, 8H, CH2NH), 3.04 (d, J = 13.1 Hz, 4H, ArCH2Ar), 2.89 (s, 8H, CH2Pbf), 2.49 (s, 12H, CH3Pbf), 2.43 (s, 12H, CH3Pbf), 2.00 (s, 12H, CH3Pbf), 1.90 -1.20 (m, 108H, OCH2CH2CH2CH2CH2, tBu and C (CH3) 2Pbf), 0.84 (t, J = 6.5 Hz, 12H, CH2CH3). <13> C NMR (75 MHz, CD3Cl3 / CD3OD = 19: 1) Î ́ 171.0, 158.6, 156.3, 155.9, 153.5, 138.2, 134.9, 132.4, 132.2, 131.0, 124.6, 121.0, 117.4, 86.4, 79.9, 75.3, 54.3, 43.0, 40.2, 31.9, 31.0, 30.0, 29.6, 28.3, 28.1 , 25.8, 25.1, 22.7, 19.1, 17.8, 13.9, 12.2. MS (ESI): calculated for [M 2Na] <++> m / z = 1449.8, found m / z = 1450.6.

Example 6 - 5,11,17,23-Tetrakis [(NÎ ± -Boc-NÏ ‰ âˆ'Pbf-Largininoyl) amino] -25,26,27,28-tetra (ethoxyethoxy) calix [4] arenas

The raw reaction obtained according to the general procedure of Example 2 is purified by preparative TLC (eluent: ethyl acetate / MeOH = 98/2) to obtain the pure product in the form of a white solid.

Melting point: 190-193 ° C. <1> H NMR (300 MHz, CD3Cl3 / CD3OD = 19: 1) Î ́ 9.05 (bs, 4H, ArNH), 6.87 (bs, 4H, ArH), 6.63 (bs , 4H, ArH), 4.35 (d, J = 13.0 Hz, 4H, ArCH2Ar), OCH2and CHCONH under the HOD, 3.70 (bs, 8H, ArOCH2), 3.50-3.35 (m, 8H, ArOCH2CH2), 3.15-2.92 ( m, 12H, CH2NH), 3.04 (d, J = 13.0 Hz, 4H, ArCH2Ar), 2.82 (s, 8H, CH2Pbf), 2.42 (s, 12H, CH3Pbf), 2.36 (s, 12H, CH3Pbf), 1.94 ( s, 12H, CH3Pbf), 1.80-1.05 (m, 88H, COCHCH2CH2, tBu, C (CH3) 2Pbf and CH2CH3). <13> C NMR (75 MHz, CD3Cl3 / CD3OD = 19: 1) Î ́ 171.1, 158.8 , 155.9, 153.1, 138.4, 134.9, 132.4, 131.4, 124.7, 121.0, 117.5, 86.5, 79.8, 73.0, 69.3, 66.2, 54.2, 42.9, 33.4, 29.4, 28.1, 27.9, 25.3, 24.6, 18.9, 17.6, 14.8 , 12.0. MS (ESI): calculated for [M 2Na] <++> m / z = 1425.7, found m / z = 1425.9.

Example 7 - 25,26,27,28-Tetrakis {3 - [(NÎ ± -Fmoc-NÏ ‰ âˆ'Mtr-L-argininoyl) amino] propoxy} calix [4] arenas

The raw reaction obtained according to the general procedure of example 2 is purified by column chromatography (eluent: CH2Cl2 / MeOH = from 100/1 to 96/4) to obtain the pure product in the form of a white solid.

Melting point: 158-160 ° C. <1> H NMR (300 MHz, CD3Cl3 / CD3OD = 19/1) Î ́ 7.95 (bs, 4H, CH2NHCO), 7.65 (d, J = 6.6 Hz, 8H, ArFmoc ), 7.44 (d, J = 6.6 Hz, 8H, ArFmoc), 7.35-7.20 (m, 8H, ArFmoc), 7.15 (t, J = 6.6 Hz, 8H, ArFmoc), 6.60-6.10 (m, 24H, ArH , ArMtr, CHNHCO and CH2NHCNH), 4.42-4.20 (m, 12H, ArCH2Ar and OCH2CHFmoc), 4.18 (bs, 4H, OCH2CHFmoc), 4.00 (bs, 4H, COCHNH), 3.93 (bs, 8H, ArOCH2), 3.74 ( s, 12H, OCH3Mtr), 3.32 (bs, 8H, OCH2CH2CH2), 3.17 (bs, 8H, CH2NHCN), 3.08 (d, J = 13.0 Hz, 4H, ArCH2Ar), 2.63 (s, 12H, CH3Mtr), 2.57 ( s, 12H, CH3Mtr), 2.10 (bs, 8H, OCH2CH2), 2.05 (s, 12H, CH3Mtr), 1.80 (bs, 8H, COCHCH2), 1.60 (bs, 8H, COCHCH2CH2). <13> C NMR (75 MHz, CD3Cl3 / CD3OD = 19: 1) Î ́ 170.7, 156.2, 154.4, 154.3, 153.7, 141.5, 141.3, 138.9, 136.1, 134.2, 132.5, 130.9, 126.0, 125.4, 124.7, 122.8, 122.6, 120.1, 117.6, 109.5, 70.3, 64.9, 53.1, 52.4, 44.7, 37.8, 35.0, 28.4, 27.9, 27.5, 23.4, 21.9, 16.1, 9.7. MS (ESI): calculated for [M 2Na] <++> m / z = 1529.6, found m / z = 1530.6.

Example 8 - 25,26,27,28-Tetrakis {3 - [(NÎ ± -Boc-NÏ ‰ −Pbf-L-argininoyl) amino] propoxy} calix [4] arenas

The raw reaction obtained according to the general procedure of example 2 is purified by column chromatography (eluent: from ethyl acetate / hexane = 7/3 to ethyl acetate / MeOH = 98/2) to obtain the pure product in the form of a white solid .

Melting point: 162-164 ° C. <1> H NMR (300 MHz, CD3CN) Î ́ 7.61 (bs, 4H, CH2NHCO), 6.66 (d, J = 6.9 Hz, 8H, ArH), 6.56 (t, J = 6.9 Hz, 4H, ArH), 6.21 (bs, 12H, NHCNHNH), 5.99 (bs, 4H, NHBoc), 4.32 (d, J = 13.1 Hz, 4H, ArCH2Ar), 4.04 (bs, 4H, COCHNH) , 3.83 (bs, 8H, OCH2), 3.36 (bs, 8H, OCH2CH2CH2), 3.15 (d, J = 13.1 Hz, 4H, ArCH2Ar), 3.13 (bs, 8H, CH2NHCNH), 2.95 (s, 8H, CH2Mtr) , 2.51 (s, 12H, CH3Pbf), 2.44 (s, 12H, CH3Pbf), 2.15 (bs, 8H, OCH2CH2), 2.02 (s, 12H, CH3Pbf), 1.75-1.20 (m, 16H, COCHCH2CH2CH2NH), 1.40 ( s, 24H, C (CH3) 2Pbf), 1.34 (s, 36H, tBu). <13> C NMR (75 MHz, CD3CN) Î ́ 173.8, 158.9, 157.1, 156.7, 156.5, 138.6, 135.6, 134.2, 132.8 , 128.9, 125.6, 122.8, 117.7, 87.2, 79.8, 73.5, 55.0, 43.2, 40.8, 37.3, 31.0, 30.7, 30.4, 26.2, 19.2, 18.0, 12.3. MS (ESI): calculated for [M 2Na] <++> m / z = 1365.7, found m / z = 1366.7.

Example 9 - 25,26,27,28-Tetrakis {3 - [(NÎ ± -Cbz-L-argininoyl) amino] propoxy} calix [4] arene, tetra-hydrochloride

The raw reaction product, obtained according to the general procedure of example 2, is triturated in ethyl ether, dissolved in MeOH and eluted on anion exchange cartridges.

<1> H NMR (300 MHz, CD3OD) Î ́ 7.96 (s, 4H, CH2NHCO), 7.42-7.20 (m, 15H, ArHCbz), 6.70-6.42 (m, 12H, ArH), 5.15-4.93 (m, 8H, CH2Cbz), 4.37 (d, J = 13.1 Hz, 4H, ArCH2Ar), 4.26-4.09 (m, 4H, COCHNH), 4.00-3.76 (m, 8H, OCH2), 3.50-3.31 (m, 8H, OCH2CH2CH2 ), 3.25-3.03 (m, 12H, ArCH2Ar and CH2NHCNH), 2.28-2.00 (m, 8H, OCH2CH2), 2.00-1.49 (m, 16H, COCHCH2CH2CH2NH). MS (ESI): calculated for [M 2Na] <++> m / z = 1835.9, found m / z = 1836.7.

Example 10 - 25,26,27,28-Tetrakis {3 - [(NÎ ± -Cbz-NÏ ‰ -Nâ € ™ Ï ‰ - (Cbz) 2-L-argininoyl) amino] propoxy} calix [4] arene

The reaction crude obtained according to the general procedure of Example 2 is purified by preparative TLC (eluent: dichloromethane / ethyl acetate = 6/1) to obtain the pure product in the form of a white solid.

<1> H NMR (300 MHz, CD3Cl3 / CD3OD = 19/1) Î ́ 7.27-6.95 (m, 60H, ArH Cbz), 6.59-6.21 (m, 12H, ArH), 5.13-4.68 (m, 24H, CH2Cbz), 4.20-3.48 (m, 24H, ArCH2Ar, COCHNH, OCH2and OCH2CH2CH2), 3.35-2.88 (m, 12H, CH2NHCNH and ArCH2Ar), 1.77 (bs, 8H, OCH2CH2), 1.49 (bs, 16H, COCH2CH2CH2). <13> C NMR (75 MHz, CD3Cl3 / CD3OD = 19: 1) Î ́ 172.3, 163.3, 160.2, 156.4, 155.4, 136.1, 134.4, 128.5, 128.2, 127.7, 123.0, 122.1, 72.3, 68.7, 66.9, 66.6 , 54.3, 48.8, 48.5, 44.2, 36.7, 30.6, 29.7, 29.5, 28.7, 24.7. MS (ESI): calculated for [M 2Na] <++> m / z = 1465.6, found m / z = 1465.4.

Example 11 - General procedure for removing Fmoc A solution of arginine calix [n] arene Fmoc protected at the upper or lower edge in anhydrous piperidine / DCM (dichloromethane) (20% v / v) is stirred at room temperature and the reaction is followed by mass spectroscopy. After completion (24 h), the reaction is poured into water, the organic phase is separated, dried over Na2SO4 anhydrous and concentrated under reduced pressure. Several times the raw material is suspended in hexane, centrifuged and the solvent above the removed solid, in order to completely eliminate both dibenzofulvene and dibenzofulvene-piperidine adducts. A used white solid is obtained without further purification.

Example 12 - 5,11,17,23-Tetrakis [NÏ ‰ -Mtr-L-argininoylamino] -25,26,27,28-tetra (n-hexyloxy) calix [4] arene

The product is obtained following the general procedure of example 11. MS (ESI): calculated for [M 3H] <+++> m / z = 765.4, found m / z = 765.9.

Example 13 - 25,26,27,28-Tetrakis {3- [NÏ ‰ -Mtr-L-argininoyl] aminopropoxy} -calix [4] arenas

The product is obtained following the general procedure of example 11. MS (ESI): calculated for [M 2Na] <++> m / z = 1085.5, found m / z = 1086.0.

Example 14 - General procedure for the removal of Pbf and Mtr A solution of calix [n] arene, functionalized at the upper or lower edge with protected Pbf and Mtr arginines, in TFA / TIS / H2O (95/5 / 5.5 ) Is stirred at room temperature. The progression of the reaction is followed using mass spectroscopy. After completion (2-24 h), the volatiles are removed under reduced pressure and the residue washed with ethyl acetate (3×) to remove excess TFA. The raw material is precipitated, washed and centrifuged with anhydrous ethyl ether (3×). The trifluoroacetate anion of the resulting TFA salts is exchanged with the addition of a 10 mM (3×) HCl solution, followed by evaporation under reduced pressure.

Example 15 - 5,11,17,23-Tetra (L-argininoylamino) -25,26,27,28-tetra (n-hexyloxy) calix [4] arene, octa-hydrochloride

The reaction crude obtained following the general procedure of example 14 is dissolved in 10 mM HCl and washed with CH2Cl2. Excess HCl is removed under reduced pressure. The crude compound is dissolved in water and TFA (0.2%), and purified by semi-preparative RP-HPLC using a C12 column. The fractions corresponding to the product are collected and the solvent is removed under reduced pressure. The residue is treated with a 10 mM (3×) HCl solution and each time the volatile substances are removed under reduced pressure in order to eliminate the TFA.

<1> H NMR (300 MHz, CD3OD) Î ́ 7.18 (s, 4H, ArH), 6.87 (s, 4H, ArH), 4.47 (d, J = 12.9 Hz, 4H, ArCH2Ar), 4.06 (bs, 4H , COCHNH3 <+>), 3.90 (t, J = 7.3 Hz, 8H, OCH2), 3.40-3.20 (m, 8H, CH2NH), 3.16 (d, J = 12.9 Hz, 4H, ArCH2Ar), 1.98 (bs, 16H, OCH2CH2and COCHCH2), 1.72 (bs, 8H, COCHCH2CH2), 1.50-1.20 (m, 24H, OCH2CH2CH2CH2CH2), 0.94 (t, J = 6.3 Hz, 12H, CH3). <1> H NMR (300 MHz, D2O ) Î ́ 7.28 (bs, 4H, ArH), 6.88 (bs, 4H, ArH), 4.30 (bs, 4H, ArCH2Ar), 4.11 (bs, 4H, COCHNH <+>

3), 3.76 (bs, 8H, OCH2), 3.40-2.89 (bs, 12 H, CH2NH and ArCH2Ar), 1.92 (bs, 16H, OCH2CH2and COCHCH2), 1.65 (bs, 8H, COCHCH2CH2), 1.34 (m, 24H , OCH2CH2CH2CH2CH2), 0.90 (t, 12H, CH3). <13> C NMR (75 MHz, CD3OD) Î ́ 167.9, 158.9, 155.1, 136.6, 136.5, 133.2, 122.6, 122.1, 77.0, 54.8, 42.2, 33.7, 32.4, 31.8, 30.2, 27.6, 25.9, 24.3, 14.8. MS (ESI): calculated for [M 2H â € “8HCl] <++> m / z = 723.5, found m / z 723.7.

Example 16 - 5,11,17,23-Tetrakis (L- argininoylamino) -25,26,27,28-tetra (n-ethoxy ethoxy) calix [4] arene octa-hydrochloride

Following the general procedure of example 14, the pure product is obtained in the form of a white solid. <1> H NMR (300 MHz, CD3OD) Î ́ 7.15 (bs, 4H, ArH), 6.98 (bs, 4H, ArH), 4.57 (d, J = 12.5 Hz, 4H, ArCH2Ar), 4.13 (bs, 8H, ArOCH2), 4.10 (bs, 4H, CHCONH), 3.94-3.85 (m, 8H, ArOCH2CH2), 3.56 (q, J = 6.9 Hz, 8H, ArOCH2CH2OCH2), CH2NH under the signal of CD3OD, 3.14 (d, J = 12.5 Hz, 4H, ArCH2Ar), 2.15-1.62 (m, 16H, COCHCH2CH2), 1.20 (t, J = 6.9 Hz , 12H, CH <1>

2CH3). H NMR (300 MHz, D2O) Î ́ 7.18 (d, J = 1.6 Hz, 4H, ArH), 7.04 (d, J = 1.6 Hz, 4H, ArH), 4.55 (d, J = 12.9 Hz, 4H, ArCH2Ar ), 4.26 (t, J = 4.8 Hz, 8H, ArOCH2), 4.14 (t, J = 6.0 Hz, 4H, CHCONH), 3.99 (t, J = 4.8 Hz, 8H, ArOCH2CH2), 3.64 (q, J = 6.9 Hz, 8H, ArOCH2CH2OCH2), 3.64 (d, J = 12.9 Hz, 4H, ArCH2Ar), 3.34-3.14 (m, 8H, CH2NH), 2.14-1.86 (m, 16H, COCHCH2CH2), 1.19 (t, J = 6.9 Hz, 12H, CH2CH3); <13> C NMR (75 MHz, CD3OD) Î ́ 167.7, 156.6, 153.4, 135.3, 131.3, 122.2, 121.9, 73.0, 69.8, 66.5, 53.2, 40.1, 30.5, 27.7, 23.1 , 14.2. MS (ESI): calculated for [M 4H â € “8HCl] <4+> m / z = 350.2, found m / z = 350.5.

Example 17 - General procedure for removing Cbz

The calix [n] arene with protected arginine is dissolved in MeOH. A catalytic quantity of Pd / C is added and the hydrogenation is carried out at 2 atm in a Parr reactor for 24 h. The progression of the reaction is followed by

<1> H NMR and ESI-MS. The catalyst is removed by filtration and the solvent is removed under reduced pressure. The residue is treated with 0.5 M HCl in MeOH up to pH 3, which is then removed in the rotavapor.

Example 18 - 25,26,27,28-Tetrakis [3- (L-argininoyl amino) propoxy] calix [4] arene, octa-hydrochloride

The product is prepared following the general procedure of example 17. If accompanied by impurities remaining after the condensation reaction between calixarene and protected arginine, the final product is obtained pure by reverse phase chromatography or by HPLC.

<1> H NMR (300 MHz, CD3OD) Î ́ 6.70-6.43 (m, 12H, ArH), 4.32 (d, J = 12.9 Hz, 4H, ArCH2Ar), 4.20-3.86 (m, 12H, COCHNH and OCH2) , 3.65-3.40 (bt, 8H, OCH2CH2CH2), 3.40-3.27 (m, 8H, CH2NHCNH), 3.18 (d, J = 12.9 Hz, 4H, ArCH2Ar), 2.35-2.10 (m, 8H, OCH2CH2), 2.10- 1.60 (m, 16H, COCHCH2CH2CH2NH). <1> H NMR (300 MHz, D2O) Î ́ 6.84 (d, J = 7.4 Hz, 8H, ArH), 6.70 (t, J = 7.4 Hz, 4H, ArH), 4.40 (d, J = 13.0 Hz, 4H, ArCH2Ar), 4.16-3.90 (m, 12H, COCHNH and OCH2), 3.72-3.55 (m, 4H, OCH2CH2CHH), 3.48-3.15 (m, 16H, OCH2CH2CHH, ArCH2Ar and CH2NHCNH), 2.40-2.15 (m, 8H, OCH2CH2), 2.10-1.85 (m, 8H, COCHCH2), 1.77-1.56 (m, 8H, COCHCH2CH2). MS (ESI): calculated for [M 2H -8HCl] <++> m / z = 639.4, found m / z = 640.0.

Example 19: Cell cultures and transient transfection assays The human Rhabdomiosarcoma RD-4 cell line, obtained from the National Cancer Institute, Frederick, Maryland, is maintained in the monolayer form using a growth medium containing 90% DMEM, 10% FBS , 2 mM l-glutamine, 100 IU / mL penicillin and 10 µg / mL streptomycin and incubated at 37 ° C in a humidified atmosphere of 95% air-5% CO2. The cells are subcultured when the state of confluence of the monolayer within the plate is between 70-90% (that is, approximately every 3-5 days). Transfections are conducted in 6-24 well plates (~ 7 cm <2> per well) at a cell confluence state of approximately 80% (approximately 3Ã — 10 <5> cells). 3 µg of plasmid and different concentrations of ligands are added to 1 ml of serum-free DMEM, mixed rapidly and incubated at room temperature for 20 minutes. Each of the obtained mixtures is carefully added to the cells after the culture medium has been removed from the cells. Formulations are prepared by adding DOPE to a plasmid-ligand mixture with a 1: 2 ligand-DOPE molar ratio, where the ligand concentration is maintained at 10 µM. A commercial product, Lipofectamine <â „¢> LTX, is used as a positive transfection control, following the manufacturer's protocol. The mixture and the cells are incubated at 37 ° C for 5 h in a humidified atmosphere of 95% air-5% CO2. Finally, the transfection mix is removed and 3 mL of growth medium is added to each transfection well and incubated for 72 h. For each well, five fields (one in the center and one for each quadrant of the well) are selected at random, without observing the cells. Transfected cells are observed for EGFP expression with a fluorescence microscope. Each experiment is performed three times. Statistical differences between treatments are calculated with Student's test and with multifactorial ANOVA.

Similar procedures are used for N2a cells, consisting of a murine neuroblastoma cell line obtained from ATCC (# CCL-131), Vero cells, consisting of a monkey kidney cell line obtained from ATCC (#CCL -81), and EADSCs cells, consisting of a primary culture of stromal cells derived from equine adipose tissue, set up in the laboratory of the inventors of the present application.

Transfection experiments can be followed by fluorescence microscopy or fluorescence-activated cell sorting (FACS) when performed using a DNA corresponding to a fluorescent protein and detecting the expression of the latter in cells, or by using the plasmids pCMVluc and pRL-TK and detecting the expression of the two corresponding luciferases inside the cell as a function of the luminescence produced by the reactions they catalyze, or by using nucleic acids labeled with fluorescent pendants.

The cytotoxicity of the studied compounds is evaluated by cell survival experiments using the MTT assay (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide). Once the Rhabdomiosarcoma cells have been treated with the calixarene derivative, the culture medium (90% DMEM, 10% FBS, 2 mM l-glutamine, and 100 IU / mL penicillin, 10 µg / mL streptomycin) containing MTT is added to them. (5 mg / mL) leaving in contact for 4 hours. Then, after the addition of an equal volume of solubilization solution (10% SDS in 0.01 M HCl), the cells are incubated overnight at 37 ° C. The specific optical density of the samples is then measured at 540 nm.

Each experiment is repeated three times and each treatment is performed with eight replicates. Statistical differences between treatments are calculated with Student's test and with multifactorial ANOVA.

The results are shown in Figures 1-7.

Figure 1 shows the transfection efficacy against Rhabdomiosarcoma (RD-4) cells (in terms of percentage of cells transfected with the pEGFP-P1 plasmid) of 5,11,17,23-Tetra (L-argininoylamino ) -25,26,27,28-tetrakis (n-hexyloxy) calix [4] arene octa-hydrochloride (LF59), in the presence and absence of the auxiliary lipid DOPE, compared to Lipofectamine LTXâ „¢ and DOPE used alone. Argininocalixarene (LF59) transfects 70% of cells without help lipid against about 50% if formulated with DOPE and 35% of LTX <TM>.

Figure 2 shows the transfection efficacy against Rhabdomiosarcoma (RD-4) cells (in terms of percentage of cells transfected with the pEGFP-P1 plasmid) of 5,11,17,23-Tetra (L-argininoylamino ) -25,26,27,28-tetrakis (n-hexyloxy) calix [4] arene octa-hydrochloride (LF59), in the presence and absence of the auxiliary lipid DOPE, compared to 25,26,27,28-tetrakis ( guanidinopropyloxy) calix [4] arene (VB6), 5,11,17,23-tetraguanidinium-25,26,27,28-tetrakis (n-propoxy) calix [4] arene (DF41), a non-macrocyclic analog of LF59 (MB13) and Lipofectamine LTXâ „¢ and DOPE used alone. Argininocalixarene (LF59) shows a higher efficiency than analogs functionalized with the guanidinium group only and a non-cyclic analogue.

Figure 3 shows the transfection efficacy against Rhabdomiosarcoma (RD-4) cells (in terms of Relative Units of Luciferase) of 5,11,17,23-Tetra (L-Arg-amino) -25, 26,27,28-tetrakis (n-hexyloxy) calix [4] arene octa-hydrochloride (LF59), in the presence and absence of the auxiliary lipid DOPE, compared to 25,26,27,28-tetrakis (guanidinopropyloxy) calix [ 4] arene (VB6), a non-macrocyclic analogue of LF59 (MB13) and to Lipofectamine LTXâ „¢ and DOPE used alone. Also in this type of experiment the superior efficiency of argininocalixarene (LF59) is confirmed compared to LTX <TM> and analogues without arginine or non macrocyclics.

Figure 4 shows the degree of cytotoxicity of 5,11,17,23-Tetra (L-Argamino) -25,26,27,28-tetrakis (n-hexyloxy) calix [4] arene octa-hydrochloride (LF59) in comparisons of human Rhabdomiosarcoma (RD-4) cells determined with experiments in the presence of MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide). Argininocalixarene (LF59) shows low toxicity in the absence of the supporting lipid and at the concentrations used for transfection.

Figure 5 shows the transfection efficacy against VERO cells (in terms of percentage of cells transfected with the pEGFP-P1 plasmid) of 5,11,17,23-Tetra (L-argininoylamino) -25,26, 27,28-tetrakis (nesyloxy) calix [4] arene octa-hydrochloride (LF59), in the presence and absence of the auxiliary lipid DOPE, compared to Lipofectamine LTXâ „¢ used alone. Argininocalixarene confirms its greater efficiency than LTX <TM> also with this cell line both 24 and 48 hours after treatment.

Figure 6 shows the transfection efficacy against N2a cells (in terms of percentage of cells transfected with the pEGFP-P1 plasmid) of 5,11,17,23-Tetra (L-argininoylamino) -25,26, 27,28-tetrakis (nesyloxy) calix [4] arene octa-hydrochloride (LF59), in the presence and absence of the auxiliary lipid DOPE, compared to Lipofectamine LTXâ „¢ used alone. Argininocalixarene confirms its greater efficiency than LTX <TM> also with this cell line both 24 and 48 hours after treatment.

Figure 7 shows the transfection efficacy against EADSCs cells (in terms of percentage of cells transfected with the pEGFP-P1 plasmid) of 5,11,17,23-Tetra (L-Arg-amino) -25, 26,27,28-tetrakis (nesyloxy) calix [4] arene octa-hydrochloride (LF59), in the presence and absence of the auxiliary lipid DOPE, compared to Lipofectamine LTXâ „¢ used alone. Argininocalixarene confirms its greater efficiency than LTX <TM> also with this cell line both 24 and 48 hours after treatment.

Claims (17)

CLAIMS 1. Compounds of general formula (I): FGFG m S. FG FG OR S OR RO S OR FG FG m m n S. FGFG m (I) in which: n is an integer from 1 to 5, preferably n is 1, 3 or 5; m is 0 or 1; each R is independently an alkyl chain containing from 1 to 12 carbon atoms optionally functionalized with hydroxyl groups, C1-C4 alkoxy groups, amino groups or aryl groups; a polyoxyethylene chain (CH2-CH2-O) pORâ € ™ in which p is an integer from 1 to 6 and Râ € ™ is hydrogen or C1-C2 alkyl; or each R is independently an alkyl chain containing from 1 to 12 carbon atoms optionally functionalized with an amidine group, or with a guanidine group or a bicyclic guanidine group having formula: H Rc N N Ra Rd N H N NOT Rb N or HH in which Ra, Rb, Rc and Rd are independently selected from hydrogen, C1-C4 alkyl, or Rc and Rd taken together with the nitrogen atom to which they are bound form a pyrrolidine, piperidine, morpholine or piperazine ring; when m = 0, S is a spacer chosen from the group consisting of -C (= O) NH (CH2) r-, -NHC (= O) (CH2) r-, -CH2NH (CH2) r-, -CH2O (CH2) r-, -CH- 2S (CH2) r-, -CH2S-S (CH2) r-, -NHC (= O) NH (CH2) r-, -NHC (= S) NH (CH2) r-, where r is zero or an integer from 1 to 3, -NHC (= O) (CH2) q- (1,2,3-triazolyl) - (CH2) râ € ™ - where q is zero or an integer from 1 to 4 and râ € ™ is an integer from 1 to 3, - (CH2) s- (1,2,3-triazolyl) - (CH2) qC (= O) NH (CH2) r- where s is zero or 1, q and r are as defined above, -C (= O) OCH2-, -CH = NOCH2-, in which 1,2,3-triazolil is a 1,2,3-triazole substituted in positions 1 and 4 or 1 and 5 from the groups indicated in the above formulas of S; the initial atom of S starting from the left is understood to be linked to FG; S is a spacer chosen from the group consisting of (-NHC (= O) CH2) 2NC (= O) (CH2) r-, (-NHC (= O) CH2) 2CHNHC (= O) (CH2) r-, (-C (= O) NHCH2CH2) 2NC (= O) (CH2) r- o (-C (= O) NH) 2CHC (= O) NH (CH2) r, where r is as defined above, when m = 1; the initial atom of S starting from the left is understood to be linked to FG; FG is a group of formula: CG CH2 q in which: q is zero or an integer from 1 to 4; CG is an amidine group, or a guanidine group or a bicyclic guanidine group having the formula: H Rc N N Ra Rd N H N NOT Rb N or HH wherein Ra, Rb, Rc and Rd are as defined above; R1 is hydrogen, an amino group, an ammonium group, an acylated amino group, a guanidine group as defined above when S is bonded to FG by means of a carbon atom; a carboxylic group or a carboxylic derivative such as a carboxylate, carboxyamide, N- (C1-C4-alkyl) carboxyamide group or N, N-bis- (C1-C4-alkyl) carboxyamide, C1-C4 alkoxycarbonyl, when S is bonded to FG by means of a nitrogen atom; their salts, enantiomers and distereomers. 2. Compounds according to claim 1, wherein R is an alkyl chain containing from 1 to 8 carbon atoms. Compounds according to claim 1 or 2, wherein R is a hexyl or octyl chain or a 2-ethoxyethoxy or 2-methoxyethoxy chain. 4. Compounds according to claims 1-3, wherein S is -C (= O) NH (CH2) r-where r is as defined in claim 1. 5. Compounds according to claims 1-4, wherein FG is a group of formula: H N NH2 CH2 q R1 wherein R1 and q are as defined in claim 1. 6. Compounds according to claims 1-5, wherein n is 1, m is 0, S is the group -C (= O) NH- and FG is a group having formula: NH H N NH3Cl CH2 q ' Cl H3N where qâ € ™ is 1, 2 or 3. 7. A compound according to claims 1-6, selected from: - 5,11,17,23-Tetrakis (L-argininoylamino) -25,26,27,28-tetra (nesyloxy) calix [4] arene octa-hydrochloride, - 5,11,17,23,29,35-Esakis (L-argininoylamino) -25,26,27,28,29,30-exa (nesyloxy) calix [6] arene dodeca-hydrochloride, - 5,11,17,23,29,35,41,47-Octakis (L-argininoylamino) -49,50,51,52,53,54,55,56-octa (n-hexyloxy) calix [6] arene exadeca-hydrochloride, - 5,11,17,23-Tetrakis (4-guanidinobutanoylamino) -25,26,27,28-tetra (nesyloxy) calix [4] arene tetra-hydrochloride, - 5,11,17,23-Tetrakis (4-guanidinobutanoylamino) -25,26,27,28-tetra (noctyloxy) calix [4] arene tetra-hydrochloride, - 5,11,17,23-Tetrakis (L-argininoylamino) -25,26,27,28-tetra (n propyloxy) calix [4] arene octa-hydrochloride, - 5,11,17,23-Tetrakis (L-argininoylamino) -25,26,27,28-tetrakis [(2-ethoxy) ethoxy] calix [4] arene octa-hydrochloride, - 5,11,17,23-Tetrakis (L-argininoylaminomethyl) -25,26,27,28-tetra (nesyloxy) calix [4] arene octa-hydrochloride, - 5,11,17,23-Tetrakis (L-argininoylaminomethyl) -25,26,27,28-tetra (noctyloxy) calix [4] arene octa-hydrochloride, - 5,11,17,23-Tetrakis [(2-amino-5-guanidinopentyl) aminomethyl] -25,26,27,28-tetra (n-hexyloxy) calix [4] arene dodeca-hydrochloride, - 5,11,17,23-Tetrakis [(2-amino-5-guanidinopentyl) aminomethyl] -25,26,27,28-tetra (n-octyloxy) calix [4] arene dodeca-hydrochloride, - 5,11,17,23-Tetrakis [di (L-argininoylamino) acetylamino] -25,26,27,28-tetra (n-hexyloxy) calix [4] arene exadeca-hydrochloride, - 5,11,17,23-Tetrakis [di (L-argininoylamino) acetylamino] -25,26,27,28-tetra (n-octyloxy) calix [4] arene exadeca-hydrochloride, 5,11,17,23 -Tetrakis [N, N-di (2-L-argininoylaminoethyl) aminocarbonyl] -25,26,27,28-tetra (nesyloxy) calix [4] arene exadeca-hydrochloride, - 5,11,17,23-Tetrakis [N, N-di (2-L-argininoylaminoethyl) aminocarbonyl] -25,26,27,28-tetra (n-octyloxy) calix [4] arene exadeca-hydrochloride. 8. Compounds of general formula (II): R1R1 S1 S1 m CF R1 R1 S1 S1 OR m CF OR RO CF OR S1 S1 R1 R1 CF no m S1S1 m R1R1 (II) in which n is an integer from 1 to 5, preferably n is 1, 3 or 5; m is 0 or 1; each R is independently an alkyl chain containing from 1 to 12 carbon atoms optionally functionalized with hydroxyl groups, C1-C4 alkoxy groups, amino groups or aryl groups; a polyoxyethylene chain (CH2-CH2-O) pORâ € ™ in which p is an integer from 1 to 6 and Râ € ™ is hydrogen or C1-C2 alkyl; or each R is independently an alkyl chain containing from 1 to 12 carbon atoms optionally functionalized with an amidine group, or with a guanidine group or a bicyclic guanidine group having formula: H Rc NN N Ra Rd H No. N O N N Rb or HH in which Ra, Rb, Rc and Rd are independently selected from hydrogen, C1-C4 alkyl, or Rc and Rd taken together with the nitrogen atom to which they are bound form a pyrrolidine, piperidine, morpholine or piperazine ring; CF is a spacer chosen from the group consisting of -C (= O) (CH2) r-, -NH (CH2) r-, where r is zero or an integer from 1 to 3, for m = 0; the initial CF atom starting from the left is understood to be linked to S1; CF is a spacer chosen from the group consisting of (-C (= O) CH2) 2NC (= O) (CH2) s-, (-C (= O) CH2) 2CHNHC (= O) (CH2) s-, (-NHCH2CH2) 2NC (= O) (CH2) s- o (- (NH) 2CHC (= O) NH (CH2) r, where s is zero or 1 and r is zero or an integer from 1 to 3, for m = 1, the initial CF atom starting from the left is understood to be linked to S1; S1 is selected from the group consisting of a divalent radical of an oligopeptide consisting of (L) and / or (D) -amino acids and / or non-chiral amino acids, containing at least one arginine or a guanidinate amino acid in which the guanidine group has formula: H Rc NN N Ra Rd H No. N O N N Rb or HH wherein Ra, Rb, Rc and Rd are as defined above; and in which S1 is linked to CF and R1 respectively through its N- and C- terminal ends, or C- and N-terminals; when S1 is linked to R1 through the N-terminal end, R1 is hydrogen, a C1-C4-acyl group, a C1-C4-alkoxycarbonyl group or a group having the formula: H Rb N N N Ra Rc O N N + or H H, when S1 is linked to R1 through the C-terminal end, R1 is a hydroxyl group or C1-C4-alkoxide or, taken together with the C-terminal carbonyl group of S1, R1 forms a carboxyamide group, N- ( C1-C4-alkyl) carboxyamide or N, N-bis- (C1-C4-alkyl) carboxyamide; their salts, enantiomers and diastereomers. 9. Compounds according to claim 8, wherein R is an alkyl chain containing from 1 to 8 carbon atoms. 10. Compounds according to claim 8 or 9, wherein R is a hexyl or octyl chain or a 2-ethoxyethoxy or 2-methoxyethoxy chain 11. A compound according to claims 8-10, selected from: - 5,11,17,23-Tetrakis (L-arginyl-glycinoylamino) -25,26,27,28-tetra (nesyloxy) calix [4] arene octa-hydrochloride, - 5,11,17,23-Tetrakis (L-arginyl-glycinoylamino) -25,26,27,28-tetra (noctyloxy) calix [4] arene octa-hydrochloride, - 5,11,17,23-Tetrakis (L-arginyl-L-argininoylamino) -25,26,27,28-tetra (nesyloxy) calix [4] arene dodeca-hydrochloride, - 5,11,17,23-Tetrakis (L-arginyl-L-argininoylamino) -25,26,27,28-tetra (noctyloxy) calix [4] arene dodeca-hydrochloride. 12. Compounds of general formula (III): FGFG m S2 OR FG S2 R2 FG <O> R2 R2 O FG m R2 S2 FG m OR n S2 FGFG m (III) in which n is an integer from 1 to 5, preferably n is 1, 3 or 5; m is 0 or 1; each R2 is independently hydrogen; an alkyl chain containing from 1 to 12 carbon atoms optionally functionalized with hydroxyl groups, C1-C4 alkoxy groups or with amino groups; or R2 is an aryl or aryloxy group or an aryl-C1-C4 alkyl chain; when m = 0, S2 is a spacer chosen from -NHC (= O) CH2-, -C (= O) NH (CH2) s (CH2CH2O) pâ € ™ (CH2) t-, -CH2NH (CH2) s (CH2CH2O) pâ € ™ (CH2) t-, -NHC (= O) NH (CH2) s (CH2CH2O) pâ € ™ (CH2) t-, -NHC (= S) NH (CH2) s (CH2CH2O) pâ € ™ (CH2) t-, where s is zero or 1, pâ € ™ is zero or an integer from 1 to 6, t is an integer from 2 to 4, - (CH2) s- (1, 2,3-triazolyl) - (CH2) tâ € ™ - where s is zero or 1 and tâ € ™ is 1 or 2, -NHC (= O) (CH2) q- (1,2,3- triazolil) - (CH2) s (CH2CH2O) pâ € ™ (CH2) t- where q is zero or an integer from 1 to 4, s is zero or 1, pâ € ™ is zero or an integer from 1 a 6, t is an integer from 2 to 4, where 1,2,3-triazolil is a 1,2,3-triazole substituted in positions 1 and 4 or 1 and 5 by the groups indicated in the formulas of S2 above reported; the initial atom of S2 starting from the left is understood to be linked to FG; S2 is a spacer chosen from (-C (= O) CH2) 2NC (= O) CH2-, (-C (= O) CH2) 2CHNHC (= O) CH2-, (-C (= O) NH) 2CHC (= O) NH (CH2) 3) - with m = 1; the initial atom of S2 starting from the left is understood to be linked to FG; FG is a group of formula: CG CH2 q R1 where q is zero or an integer from 1 to 4; CG is an amidine group, or a guanidine group or a bicyclic guanidine group having the formula: Rc N N N Ra Rd H No. N O N N Rb or HH in which Ra, Rb, Rc and Rd are independently selected from hydrogen, C1-C4 alkyl, or Rc and Rd taken together with the nitrogen atom to which they are bound form a pyrrolidine, piperidine, morpholine or piperazine ring; R1 is hydrogen, an amino group, an ammonium group, an acylated amino group, a guanidine group as defined above, a carboxylic group or a carboxylic derivative such as a carboxylate, carboxyamide, N- (C1-C4-alkyl) carboxyamide group, N, N-bis- (C1-C4-alkyl) carboxyamide or C1-C4 alkoxycarbonyl, provided that, when S2 is bonded to FG by a nitrogen atom, R1 is hydrogen or a carboxylic derivative as defined above; their salts, enantiomers and diastereomers. 13. Compounds according to claim 12, wherein R2 is a C6-C8 alkyl or benzyl or C6H5 chain. 14. A compound according to claim 12 or 13, selected from: - 5,11,17,23-Tetra-n-hexyl-25,26,27,28-tetrakis [3- (N-L-argininoylamino) propoxy] calix [4] arene octa-hydrochloride, - 5,11,17,23-Tetra-n-hexyl-25,26,27,28-tetrakis [N, N-di (L-argininoylmethyl ester) carbonylmethyl] -3-aminopropoxyx [4] arene octachlorhydrate, - 5,11,17,23-Tetra-n-hexyl-25,26,27,28-tetrakis {N, N-di [2- (L-argininoylamino) ethyl] aminocarbonylmethoxy} calix [4] arene exadeca-hydrochloride . 15. Compounds according to claims 1-14, for use as non-viral vectors for cell transfection. 16. Compositions comprising at least one compound of formula (I) or (II) or (III) according to claims 1-14 in a suitable vehicle. 17. Use of a compound according to claims 1-14 for the preparation of a composition for cell transfection.