Dihydropyrene derivatives, processes for preparing the same and their uses

Abstract

Disclosed are dihydropyrene derivatives, processes for preparing the same and their uses.

Claims

1. A method for treating pathologies sensitive to singlet oxygen, in particular for phototherapy and/or for treating cancers comprising administering to a subject in need thereof a compound of the following formula I ##STR00220## wherein: n represents 0 or 1, represents a single bond or no bond, represents a single or a double bond, R.sub.1 and R.sub.1 represent independently from each other: H, a linear or branched (C.sub.1-C.sub.18)-alkyl, a (C.sub.3-C.sub.8)-cycloalkyl, ##STR00221## N(R).sub.3.sup.+, R representing: H, a linear or branched (C.sub.1-C.sub.18)-alkyl, a (C.sub.3-C.sub.8)-cycloalkyl, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 represent independently from each other: H, a linear or branched (C.sub.1-C.sub.18)-alkyl, a (C.sub.3-C.sub.8)-cycloalkyl, ##STR00222## N(R).sub.3.sup.+, R representing: H, a linear or branched (C.sub.1-C.sub.18)-alkyl, a (C.sub.3-C.sub.8)-cycloalkyl, X represents one or more physiologically acceptable counter anion(s), providing that: at least one of R.sub.1, R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 represents one of the following groups: ##STR00223## or N(R).sub.3.sup.+, n, and the bonds 1 to 7 are such as: n=0, represents a single bond, bonds 1, 4 and 6 represent a double bond, and bonds 2, 3, 5 and 7 represent a single bond, or n=0, represents no bond, bonds 1, 3, 5 and 7 represent a double bond, and bonds 2, 4 and 6 represent a single bond, or n=1, represents no bond, bonds 2, 5 and 7 represent a double bond, and bonds 1, 3, 4 and 6 represent a single bond.

2. The method according to claim 1 wherein: R.sub.1 and R.sub.1 are identical; R.sub.1 and/or R.sub.1 represent(s) a linear or branched (C.sub.1-C.sub.18)-alkyl, in particular a tert-butyl; R.sub.2 and/or R.sub.4 represent(s) ##STR00224## R.sub.3 and R.sub.5 representing in particular H; R.sub.3 and/or R.sub.5 represent(s) ##STR00225## R.sub.2 and R.sub.4 representing in particular H; or X is (are) chosen from the group consisting in Cl.sup., PF.sub.6.sup., BF.sub.4.sup., CH.sub.3COO.sup., Br.sup., F.sup., SO.sub.4.sup.2, HSO.sub.4.sup., HPO.sub.4.sup.2, H.sub.2PO.sub.4.sup.; or R represents a linear or branched (C.sub.1-C.sub.18)-alkyl, in particular CH.sub.3.

3. The method according to claim 1 wherein the compound is one of the compound of the following formulae: ##STR00226## ##STR00227##

4. The method according to claim 1, wherein the compound of formula I forms a complex of the following formula II ##STR00228## B group being chosen from a peptide or an acid residue selected from a hyaluronic acid or a folic acid.

5. A pharmaceutical or diagnostic composition comprising a compound of formula I according to claim 1 as active agent and a pharmaceutically acceptable vehicle.

6. Compound of the following formula I (bis) ##STR00229## wherein: n is 1, represents a single bond or no bond, represents a single or a double bond, R.sub.1 and R.sub.1 represent independently from each other: H, a linear or branched (C.sub.1-C.sub.18)-alkyl, a (C.sub.3-C.sub.8)-cycloalkyl, ##STR00230## N(R).sub.3.sup.+, R representing: H, a linear or branched (C.sub.1-C.sub.18)-alkyl, a (C.sub.3-C.sub.8)-cycloalkyl, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 represent independently from each other: H, a linear or branched (C.sub.1-C.sub.18)-alkyl, a (C.sub.3-C.sub.8)-cycloalkyl, ##STR00231## N(R).sub.3.sup.+, R representing: H, a linear or branched (C.sub.1-C.sub.18)-alkyl, a (C.sub.3-C.sub.8)-cycloalkyl, X represents one or more counter anion(s), in particular one or more physiologically acceptable counter anion(s), providing that: at least one of R.sub.1, R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 represents one of the following groups: ##STR00232## or N(R).sub.3.sup.+.sub.f n, and the bonds 1 to 7 are such as: n=1, represents no bond, bonds 2, 5 and 7 represent a double bond, and bonds 1, 3, 4 and 6 represent a single bond.

7. Compound according to claim 6, wherein: R.sub.1 and R.sub.1 are identical; R.sub.1 and/or R.sub.1 represent(s) a linear or branched (C.sub.1-C.sub.18)-alkyl, in particular a tert-butyl; R.sub.2 and/or R.sub.4 represent(s) ##STR00233## R.sub.3 and R.sub.5 representing in particular H; R.sub.3 and/or R.sub.5 represent(s) ##STR00234## R.sub.2 and R.sub.4 representing in particular H; X is (are) chosen from the group consisting in Cl.sup., PF.sub.6.sup., BF.sub.4.sup., CH.sub.3COO.sup., Br.sup., F.sup., SO.sub.4.sup.2, HSO.sub.4.sup., HPO.sub.4.sup.2, H.sub.2PO.sub.4.sup.; or R represents a linear or branched (C.sub.1-C.sub.18)-alkyl, in particular CH.sub.3.

8. Compound according to claim 7, of one of the following formulae: ##STR00235##

9. A pharmaceutical or diagnostic composition comprising a compound of formula I according to claim 1 as active agent and a pharmaceutically acceptable vehicle.

10. A pharmaceutical or diagnostic composition comprising a compound of formula I according to claim 2 as active agent and a pharmaceutically acceptable vehicle.

11. A pharmaceutical or diagnostic composition comprising a compound of formula I according to claim 3 as active agent and a pharmaceutically acceptable vehicle.

12. A pharmaceutical or diagnostic composition comprising a complex of formula II according to claim 4 as active agent and a pharmaceutically acceptable vehicle.

13. The method according to claim 4, wherein the B group is a peptide of the following formula: ##STR00236##

14. A compound of the following formula I (bis) ##STR00237## wherein: n represents 0 or 1, represents a single bond or no bond, represents a single or a double bond, R.sub.1 and R.sub.1 represent independently from each other: H, a linear or branched (C.sub.1-C.sub.18)-alkyl, a (C.sub.3-C.sub.8)-cycloalkyl, ##STR00238## N(R).sub.3.sup.+, R representing: H, a linear or branched (C.sub.1-C.sub.18)-alkyl, a (C.sub.3-C.sub.8)-cycloalkyl, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 represent independently from each other: H, a linear or branched (C.sub.1-C.sub.18)-alkyl, a (C.sub.3-C.sub.8)-cycloalkyl, ##STR00239## N(R).sub.3.sup.+, R representing: H, a linear or branched (C.sub.1-C.sub.18)-alkyl, a (C.sub.3-C.sub.8)-cycloalkyl, X represents one or more counter anion(s), in particular one or more physiologically acceptable counter anion(s), providing that: at least one of R.sub.1, R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 represents one of the following groups: ##STR00240## or N(R).sub.3.sup.+, n, and the bonds 1 to 7 are such as: n=0, represents a single bond, bonds 1, 4 and 6 represent a double bond, and bonds 2, 3, 5 and 7 represent a single bond, or n=0, represents no bond, bonds 1, 3, 5 and 7 represent a double bond, and bonds 2, 4 and 6 represent a single bond, or n=1, represents no bond, bonds 2, 5 and 7 represent a double bond, and bonds 1, 3, 4 and 6 represent a single bond, when n=0, R.sub.1 and/or R.sub.1 are different from one of the following groups: ##STR00241## or N(R).sub.3.sup.+, wherein the compound of formula I(bis) forms a complex of the following formula II(bis) ##STR00242## II (bis) B group being chosen from a peptide or an acid residue selected from a hyaluronic acid or a folic acid.

15. The compound according to claim 14, wherein: R.sub.1 and R.sub.1 are identical; R.sub.1 and/or R.sub.1 represent(s) a linear or branched (C.sub.1-C.sub.18)-alkyl, in particular a tert-butyl; R.sub.2 and/or R.sub.4 represent(s) ##STR00243## R.sub.3 and R.sub.5 representing in particular H; R.sub.3 and/or R.sub.5 represent(s) ##STR00244## R.sub.2 and R.sub.4 representing in particular H; X is (are) chosen from the group consisting in Cl.sup., PF.sub.6.sup., BF.sub.4.sup., CH.sub.3COO.sup., Br.sup., F.sup., SO.sub.4.sup.2, HSO.sub.4.sup., HPO.sub.4.sup.2, H.sub.2PO.sub.4.sup.; or R represents a linear or branched (C.sub.1-C.sub.18)-alkyl, in particular CH.sub.3.

16. The compound according to claim 14, of one of the following formulae: ##STR00245## ##STR00246##

17. The compound according to claim 14, wherein the B group is a peptide of the following formula: ##STR00247##

Description

FIGURES

(1) FIG. 1 represents the .sup.1H-NMR spectra of compound 1 in CD.sub.3CN.

(2) FIG. 2 represents the .sup.1H-NMR spectra of compound 2 in CD.sub.3CN.

(3) FIG. 3 represents the .sup.1H-NMR spectra of compound 3 in CD.sub.3CN.

(4) FIG. 4 represents the UV-visible absorption spectra of 1 (solid line), 2 (dotted line) and 3 (diamond line) in CH.sub.3CN.

(5) FIG. 5 presents the .sup.1H NMR spectra of: (A) 1, (B) Photogenerated 3, and (C) reaction mixture in CD.sub.3CN after cyeloreversion of 3 back to 1 in the presence of 30-fold excess 2,3-dimethyl-2-butene. Signals at 4.88 and 4.93 ppm in C correspond to the two olefin protons for trapped product.

(6) FIG. 6 presents the EPR spectra of nitroxide radical generated from TEMPD. 1M solution of TEMPD was mixed with 44 M of 1. A: initial state. B: upon irradiation with red light (>630 nm). C: upon thermal relaxation and recovery of 1.

(7) FIG. 7 presents the evaluation of the cytotoxicity of compound 5 under irradiation on IGROV1 cells. The graph corresponds to the viability of the cells, in percent, in function of the concentration of compound 5 in M.

(8) FIG. 8 represents the flow cytometer analysis of compound 5. The dark grey line corresponds to the IGROV1 cells without addition of a compound of the Invention, the Mack line corresponds to the IGROV1 cells incubated with compound 4 and the grey line corresponds to the IGROV1 cells incubated with complex 5.

EXAMPLES

(9) General Procedures and Methods

(10) All purchased chemicals and solvents were used as received except THF and diethyl ether that were distillated over sodium/benzophenone under argon. NMR spectra were recorded on a Bruker Avance-500 MHz or 400 MHz spectrometer in CD.sub.3CN. Chemical shifts (ppm) are referenced to residual solvent peaks. Mass spectrometry analyses (ESI positive mode) were carried out at the DCM mass spectrometry facility with an Esquirre 3000 Plus (Bruker Daltonics). Absorption spectra were recorded using either a Varian Cary 50 Scan or a Varian Cary 300 UV-visible spectrophotometer equipped with a temperature controller unit. Luminescence spectra in the NIR were recorded on an Edinburgh Instruments FLS-920 spectrometer equipped with a Ge detector cooled at 77K.

(11) Irradiation experiments have been conducted either under inert atmosphere using a Jaram glove box with carefully degassed solvents or under air (1 atm). Visible irradiations experiments have been carried out with a XeHg lamp, using a 630 nm cut-off filter unless otherwise stated and the samples have been placed in a water bath (room temperature or 8 C.). Samples have been placed at a distance of 15 cm of the visible lamp. The reactions have been investigated from UV-visible and NMR experiments. Intermediate spectra have been recorded at different times depending on the isomerization process rates. The ratio between the different forms has been determined by .sup.1H-NMR from the relative integration of the characteristic resonance peaks of the N.sup.+-Me methyl groups of the different forms.

Example 1: Preparation of compound 1: 2,7-di-tert-butyl-4,9-di-(N-methylpyridin-4-yl)-trans-10b,10c-dimethyl-10b,10c-dihydropyrene hexafluorophosphate

(12) 1 was prepared as represented in Scheme S1.

(13) ##STR00210##

(14) 2,7-Di-tert-butyl-trans-10b,10c-dimethyl-10b,10c-dihydropyrene (DHP), and 4,9-dibromo-2,7-di-tert-butyl-trans-10b,10c-dimethyl-10b-10c dihydropyrene (DRP-Br.sub.2) were synthesized following the procedures described in Mitchell et al. (J. Am. Chem. Soc. 2003, 125, 2974-2988) and Vila et al. (Inorg. Chem. 2011, 50, 10581-10591).

2,7-di-tert-butyl-4,9-di-(4-pyridyl)-trans-10b,10c-dimethyl-10b,10c-dibydropyrene (DHP-Py2)

(15) A round bottom flask was filled under an argon atmosphere with 4,9-dibromo-2,7-di-tert-butyl-trans-10b,10c-dimethyl-10b,10c-dihydropyrene (0.100 g, 0.2 mmol), 4-pyridinylboronic acid (52 mg, 0.42 mmol) and freshly distilled THF (6 mL). A degassed aqueous solution (2 mL) of sodium carbonate (0.1 g, 0.95 mmol) and Pd(PPh.sub.3).sub.4 (35 mg, 0.03 mmol) was then added into the flask and the resulting mixture was heated under stirring for 48 h. After cooling the mixture to room temperature, the solvent was evaporated under reduced pressure and the residue was then dissolved in water and extracted with dichloromethane. The solution was dried over anhydrous MgSO.sub.4, filtered and the solvent was evaporated under reduced pressure. Around 5 mL of diethyl ether were added to the crude product and the insoluble DHP-Py.sub.2 was then filtered and dried under vacuum. DHP-Py.sub.2 was isolated as a dark brown solid (40 mg, 40% yield). DHP-Py.sub.2: .sup.1H NMR (400 MHz, 298 K, CDCl.sub.3) (ppm): 3.68 (s, 6H), 1.61 (s, 18H), 7.78 (m, 4H), 8.48 (s, 2H), 8.60 (s, 2H), 8.64 (s, 2H), 8.85 (m, 4H). ESIMS: m/z: calcd for C.sub.36H.sub.38N.sub.2+H.sup.+: 499.7 [M+H.sup.+] found: 499.4. Exact mass (M.sup.+) calc.: 499.3107, found: 499.3105. Anal. Calc. for C.sub.36H.sub.38N.sub.2.0.5H.sub.2O: C, 85.16; H, 7.74; N, 5.52. found: C, 85.16; H, 7.96; N, 5.10. 5o: RMN 1H (400 MHz, 298 K, CDCl.sub.3) (ppm): 1.18 (s, 18H), 1.54 (s, 6H), 6.57 (d, J=2 Hz, 2H), 6.84 (s, 2H), 6.86 (d, J=2 Hz, 2H), 7.48 (m, 4H), 8.61 (m, 4H).

2,7-di-tert-butyl-4,9-di-(N-methylpyridin-4-yl)-trans-10b,10c-dimethyl-10b,10c-dihydropyrene hexafluorophosphate (1, 2 PF6)

(16) 35 mg of DHP-Py.sub.2 (0.070 mmol) were dissolved in 20 mL CH.sub.2Cl.sub.2. 1 mL of CH.sub.3I was then rapidly added and the solution was refluxed for two hours. Upon cooling down to room temperature, the precipitate formed (iodide salt) was filtered off, washed with cold CH.sub.2Cl.sub.2 and dissolved in 40 mL CH.sub.3OH. Addition of a saturated aqueous solution of NH.sub.4PF.sub.6 precipitated the hexafluorophosphate salt of 1 as a red-brown solid that was collected by filtration, washed with cold water and CH.sub.3OH and dried under vacuum. Crystals could be obtained by slow diffusion of diethyl ether into a CH.sub.3CN solution of 1 (yield 92%, 45 mg, 64.4 mol). 1: .sup.1H NMR (500 MHz, 298 K, CD.sub.3CN) (ppm): 3.63 (s, 6H), 1.65 (s, 18H), 4.43 (s, 6H), 8.45 (m, 4H), 8.70 (s, 2H), 8.73 (s, 2H), 8.79 (m, 4H), 8.89 (s, 2H). Exact mass: m/z: calcd for C.sub.38H.sub.44N.sub.2.sup.2+: 264.1747 [M-2PF.sub.6.sup.], found: 264.1750.

Example 2: Preparation of compound 4: 2,7-di-tert-butyl-4-(N-methylpyridin-4-yl)-trans-10b,10c-dimethyl-10,10c-dihydropyrene hexafluorophosphate

(17) ##STR00211##

4-bromo-2,7-di-tert-butyl-trans-10b,10c-dimethyl-10b-10c-dihydropyrene

(18) ##STR00212##

(19) To a solution containing 2,7-di-tert-butyl-trans-10b,10c-dimethyl-10b,10c-dihydropyrene (350 mg, 1.02 mmol) in 190 mL of dry CH.sub.2Cl.sub.2 at 40 C. was slowly added (1 hour) with stirring under an argon atmosphere a solution containing N-bromosuccinimide (181 mg, 1.02 mmol) in dry DMF (35 mL) at 40 C. After addition, the solution was kept under stirring 1 hour at room temperature. Cyclohexane (80 mL) and water were then added. The organic phase was collected, washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and evaporated under reduced pressure. The crude product was purified by column chromatography on silica gel using cyclohexane:CH.sub.2Cl.sub.2 (6:1, vol:vol) as eluent to afford 4-bromo-2,7-di-tert-butyl-trans-10b,10c-dimethyl-10b-10c-dihydropyrene as dark green crystals. (411 mg, yield 90%) .sup.1H NMR (400 MHz, CDCl.sub.3) /ppm: 8.81 (d, 1H), 8.64 (s, 1H), 8.54 (d, 2H, J=1.7 Hz), 8.48 (bs, 1H), 8.47 (s, 2H), 1.71 (s, 9H, t-Bu), 1.68 (s, 9H, t-Bu), 3.91 (s, 3H, CH.sub.3), 3.92 (s, 3H, CH.sub.3).

2,7-di-tert-butyl-4-(4-pyridyl)-trans-10b,10c-dimethyl-10b,10c-dihydropyrene

(20) ##STR00213##

(21) 4-bromo-2,7-di-tert-butyl-trans-10b,10c-dimethyl-10b,10c-dihydropyrene (0.100 g, 0.236 mmol) and 4-pyridinylboronic acid (32 mg, 0.26 mmol) were dissolved in degassed and freshly distilled THF (6 mL). A solution of sodium carbonate (100 mg, 0.95 mmol) in water (2 mL) and tetrakis(triphenylphosphine)palladium(0) (35 mg, 0.03 mmol) were then introduced under an inert atmosphere. The mixture was refluxed for 48 h. The suspension was then cooled down to room temperature. The solvent was evaporated to dryness under reduced pressure. The solid residue was washed with water and extracted with CH.sub.2Cl.sub.2 (320 mL). The organic phases were collected and combined, dried over anhydrous MgSO.sub.4 and evaporated under reduced pressure. The crude product was purified by column chromatography on silica gel using cyclohexane as eluent. After having eluted the first fraction which was the unreacted monobromide derivatives, the polarity of the eluent was gradually increased up to cyclohexane:ethyl acetate 50:50 (vol:vol) allowing to afford 50 mg of compound as a dark brown solid (yield 80%, 0.21 mmol). 3.sub.c: .sup.1H NMR (400 MHz, 298 K, CDCl.sub.3) (ppm): (400 MHz, 298 K, CDCl3) d (ppm): 3.84 (s, 3H), 3.83 (s, 3H), 1.72 (s, 9H), 1.64 (s, 9H), 7.77 (dd, 2H), 8.46 (s, 1H), 8.48 (s, 2H), 8.56 (m, 2H), 8.58 (s, 1H), 8.62 (s, 1H), 8.83 (dd, 2H). Exact mass: m/z: calcd for C.sub.31H.sub.35N +H.sup.+: 422.2841 [M+H.sup.+], found: 422.2841. 3.sub.o: .sup.1H NMR (400 MHz, 298 K, CDCl.sub.3) (ppm): 1.19 (s, 9H), 1.25 (s, 9H), 1.47 (s, 3H), 1.50 (s, 3H), 6.40 (dd, 2H), 6.52 (d, 2H), 6.73 (br, 1H), 6.78 (br, 4H), 7.45 (dd, 2H).

2,7-di-tert-butyl-4-(N-methylpyridin-4-A-trans-10b,10c-dimethyl-10b,10c-dihydropyrene hexafluorophosphate

(22) ##STR00214##

(23) 20 mg of 2,7-di-tert-butyl-4-(4-pyridyl)-trans-10b,10c-dimethyl-10b,10c-dihydropyrene (0.047 mmol) were dissolved in 20 mL diethyl ether. 1 mL of CH.sub.3I was then rapidly added and the solution was refluxed for 4 hours. Upon cooling down to room temperature, the precipitate formed (iodide salt) was filtered off, washed with diethyl ether and dissolved in 20 mL CH.sub.3OH. The hexafluorophosphate salt was then precipitated upon addition of a saturated aqueous solution of NH.sub.4PF.sub.6. The orange powder was then collected by filtration, washed with cold water and dried under vacuum (yield 89%, 22 mg, 42 mol). .sup.1H NMR (500 MHz, 298 K, CD.sub.3CN) (ppm): 3.88 (s, 3H), 3.84 (s, 3H), 1.65 (s, 9H), 1.70 (s, 9H), 4.39 (s, 3H), 8.43 (d, 2H), 8.61 (d, 1H), 8.63 (s, 1H), 8.65 (d, 1H), 8.71 (s, 1H), 8.73 (d, 2H), 8.77 (s, 1H), 8.78 (s, 1H), 8.80 (s, 1H). Exact mass: m/z: calcd for C.sub.32H.sub.38N.sup.+: 436.2999 [M-PF.sub.6.sup.], found: 436.2998. .sup.1H NMR (500 MHz, 298 K, CD.sub.3CN) (ppm): 1.20 (s, 9H), 1.25 (s, 9H), 1.41 (s, 3H), 1.48 (s, 3H), 4.23 (s, 3H), 6.45 (d, 1H), 6.49 (d, 1H), 6.56 (s, 1H), 6.90 (s, 1H), 6.94 (s, 1H), 6.94 (s, 1H), 7.29 (s, 1H), 8.03 (d, 2H), 8.44 (d, 2H).

(24) Irradiation Procedures

(25) Samples for experiments under inert atmosphere were prepared in a Jaram glove box with carefully degassed solvents, or were thoroughly purged with argon. Solutions for experiments in the presence of oxygen were prepared under air (1 atm). The solutions were irradiated in UV-visible quartz cells or NMR tubes. The concentration used for UV-visible spectroscopy and NMR experiments were comprised between 210.sup.5 M and 310.sup.3 M. The visible irradiations for making the isomerization of the closed 1 isomer to its corresponding open 2 were carried out with a XeHg lamp, using a 630 nm cut-off filter and the samples were placed at 8 C. bath in order to limit the reverse thermal reaction. Samples were placed at a distance of 15 cm of the visible lamp. Alternatively, irradiation was performed at room temperature with a 150 W tungsten-halogen lamp equipped with a 590 nm cut-off filter. The conversions between the different species were investigated from UV-visible and NMR experiments. Intermediate spectra were recorded at different times depending on the isomerization processes rates. The ratio between the different species was determined by .sup.1H-NMR from the relative integration of the characteristic resonance peaks of the N.sup.+CH.sub.3 groups of the different forms.

(26) ##STR00215##

Example 3: Synthesis of Compound 2

(27) 2 was generated by visible irradiation of a solution of 1 under inert following the procedure described above.

(28) 2: .sup.1H NMR (500 MHz, 298 K, CD.sub.3CN) (ppm): 1.20 (s, 18H), 1.50 (s, 6H), 4.26 (s, 6H), 6.69 (s, 4H), 7.12 (s, 2H), 7.37 (s, 2H), 8.06 (m, 4H), 8.49 (m, 4H). Mass (m/z): calcd: 673.3 [M-PF.sub.6.sup.], found: 673.3.

Example 4: Synthesis of Compound 3

(29) 3 was generated by visible irradiation of a solution of 1 under air following the procedure described above.

(30) 3: .sup.1H NMR (500 MHz, 298 K, CD.sub.3CN) (ppm): 0.05 (s, 3H), 1.04 (s, 9H), 1.24 (s, 9H), 2.07 (s, 3H), 4.24 (s, 3H), 4.28 (s, 3H), 6.51 (s, 1H), 7.00 (s, 1H), 7.06 (d, J=2.1 Hz, 1H), 7.22 (d, J=2.1 Hz, 1H), 7.42 (d, J=2.1 Hz, 1H) 7.64 (s, 1H) 7.88 (d, J=7.0 Hz, 2H), 8.07 (d, J=7.0 Hz, 2H), 8.49 (d, J=7.0 Hz, 2H), 8.54 (3=7.0 Hz, 2H). Mass (m/z): calcd: 705.3 [M-PF.sub.6.sup.], found: 705.3.

Example 5: Phosphorescence Measurements

(31) Luminescence measurements in the near infrared (NIR) region were performed with an Edinburgh Instruments FLS-920 spectrometer equipped with a germanium detector cooled with liquid nitrogen. The luminescence was recorded on air-equilibrated CD.sub.3CN solutions contained in 3-mL quartz cells with 1-cm path length. Deuterated acetonitrile was used as the solvent to increase the sensitivity of the measurement, owing to the significantly higher emission quantum yield of singlet oxygen in comparison with CH.sub.3CN. Compound 3 was generated in situ by exhaustive irradiation of 1 in the visible region (ca. 60 min irradiation under the conditions employed) at room temperature. The solution was then warmed up at 60 C. and its luminescence properties (spectral dispersion and time dependence of the intensity) were monitored in the absence of photoexcitation (the excitation source of the spectrometer was turned off).

Example 6: 1H NMR Analysis of Singlet Oxygen Trapping

(32) Singlet oxygen trapping experiments were carried out by NMR using 2,3-dimethyl-2-butene.

(33) ##STR00216##

(34) A solution of 1 (3 mM) in CD.sub.3CN was irradiated (>630 nm) and converted into 3. A 30 fold excess (90 mM) of 2,3-dimethyl-2-butene was added to the solution and the sample was maintained at 35 C. in the dark. After 48 h, a .sup.1H NMR spectra was acquired with simultaneous saturation of the large signal at 1.8 ppm due to unreacted 2,3-dimethyl-2-butene. The two olefin proton signals for trapped hydroperoxide appear in the open window of 4.88-4.93 ppm. Comparison of the average peak integrals with signals of regenerated 1 showed that 8510% of the released singlet oxygen was trapped (FIG. 5).

Example 7: Evidence of Singlet Oxygen Production by Trapping ESR Experiments

(35) Singlet oxygen trapping experiments were carried out by ESR using 2,2,6,6-tetramethyl-4-piperidone, TEMPD (Hideg et al. Biochim. Biophys. Acta 2011, 1807, 1658-1661).

(36) A solution of 1 (44 M) with TEMPD (1M) in CH.sub.3CN was irradiated (>630 nm) and converted into 3. ESR spectra were recorded before and after irradiation process (FIG. 6). The sample was then heated in the dark to release singlet oxygen.

Example 8: In Vitro Experiments

(37) Compounds of formula I-3 are incubated in the presence of cells and cell viability is observed by microscopy after heating 48 h at 37 C.

(38) Compounds of formula I-1 are incubated in the presence of cells and cell viability is observed: after heating 48 h at 37 C. (control); or after irradiation at 630 nm in presence of oxygen, and then heating 48 h at 37 C.

(39) Compounds of formula I-2 are incubated in the presence of cells and cell viability is observed: after heating 48 h at 37 C. (control); or after contacting said compounds of formula I-2 with singlet oxygen, said singlet oxygen being generated beforehand or in situ, in presence of an external photosensitizer and oxygen or an oxygen containing gas, and then heating 48 h at 37 C.

Example 9: Preparation of Complex 5

(40) The homodetic cyclopeptide Raft(4GRD)CysNPys was synthesized according to the FR 02 11614 patent.

(41) Synthesis of the DHP-SH

(42) ##STR00217##

(43) Synthesis of DHP-SCOCH.sub.3

(44) DHP-Py (45 mg/0.1 mmol) and 3 equivalents of bromo-thioacetatealkyl A (85 mg/0.3 mmol) are dissolved in 20 mL of CH.sub.3CN. The solution is then refluxed during one week under inert atmosphere. Upon cooling, the solution is then concentrated under vacuum and the product (DHP-SCOCH.sub.3) is precipitated by addition of diethyl ether. The red-brown solid is filtered, washed with diethyl ether and dried under vacuum. Yield: 76%.

(45) Synthesis of DHP-SH

(46) The protected compound DHP-SCOCH.sub.3 (20 mg/0.03 mmol) is dissolved in MeOH (2 mL) and cone. HCl (0.5 mL) under inert atmosphere and the mixture is then refluxed for 3 hours. Upon cooling, solvents are removed under vacuum to afford the deprotected compound DHP-SH in quantitative yield. This product can be used without further purifications.

(47) Vectorization

(48) ##STR00218##

(49) 10 mg of Raft(4RGD)CysNPyS (2.41 mol, 1 eq.) in 600 L of DMF/H.sub.2O/CH.sub.3CN (1/1/1) are submitted to three empty/Argon cycles. The DHP-SH (3 mg, 4.82 mol, 2 eq.) is dissolved in 600 L of CH.sub.3CN/Acetate buffer (pH=5.2, 100 mM) (2/1). This solution is protected from light and submitted to 3 empty/Argon cycles. The Raft(4RGD)CysNPyS and DHP-SH solutions are mixed and protected from light. After three empty/Argon cycles, the reaction mixture is stirred 2 h at room temperature. The crude is purified by RP-HPLC to afford the final product 5 as a brown powder (1.5 mg, 332 nmol) Yield: 14%; RP-HPLC: RT=17 min (gradient 10% to 85% CH.sub.3CN in 20 min; column 100-7 C18, 214 nm). MS: calcd for C.sub.211H.sub.304N.sub.57O.sub.51S.sub.2 MW 4519.25 g.mol.sup.1. Found MW=4518.4 g.mol.sup.1.

(50) Irradiation (According to the General Procedure):

(51) The visible irradiation induces the isomerization of the closed 5 isomer to its corresponding open form 6 when performed in the absence of oxygen. In the presence of oxygen, the visible irradiation of the closed isomer 5 produces the corresponding endoperoxide open form 7.

(52) ##STR00219##

Example 10: Evaluation of the Cytotoxicity of Complex 5

(53) The ability of complex 5 to induce the death of cancerous cells under near infrared irradiation has been evaluated on human ovarian adenocarcinoma IGROV1, using the MTT test. IGROV1 cells were cultivated in a 10% SVF supplemented RPMI-1640 medium and incubated at 37 C. under a 5% CO.sub.2 atmosphere.

(54) The cells were seeded on a 96-well plate with a density of 5000 cells per well. Twenty four hours later, the culture medium was removed and replaced by a fresh medium containing different concentrations of complex 5 comprised from 1 to 100 m (n=3 wells per conditions). The culture plates were irradiated with a 2.4 mW/cm.sup.2 680 nm laser for 85 minutes at 37 C. 48 hours later, the cytotoxicity is determined using a MTT test provided by Sigma-Aldrich (3-(4,5-di-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). The MTT solution (0.2 mg/mL in PBS) is added to the wells (100 L/well) and the plates were incubated 2 hours at 37 C. Formazan crystals that Mimed were then dissolved with 100 L/well propanol and the absorbance at 570 nm was read on an absorbance reader (AD 340, Beckman counter). The results are expressed in percentage of the mean value obtained with the untreated wells (FIG. 7).

(55) This cellular culture experiment highlights the capacity of complex 5 to induce the death of cancerous cells under a near infrared irradiation. At a 50 M concentration, the death of 50% of the cells is observed.

Example 11: Ability to Target the Cancerous Cells

(56) The ability of compounds 4 and 5 to cross the ovarian adenocarcinoma cell (IGROV1) membrane or to fix the cell membrane of these cells were evaluated by flow cytometry. The cells were seeded in Petri dishes at a 10.sup.6 density of cells per dish. Twenty four hours later, the culture medium was removed and replaced by a fresh medium containing compound 4 or compound 5 at a 50 M concentration. After 2 hours of incubation at 37 C., the cells were rinsed with PBS, trypsinized, centrifuged and then suspended with a concentration of 10.sup.6 cells per mL of PBS. The fluorescence of the cells was observed with a flow cytometer (Accuri C6, BD) at a 488 nm excitation wavelength and with a 680 nm high-pass emission filter.

(57) FIG. 8 presents the results of the flow cytometry analysis, showing the significative enhancement of the ability of complex 5 to cross or to fix the cell membrane compared to its unvectorized analog compound 4.

Example 12: In Vitro Experiments with Complex of Formula II

(58) Experiments according to example 8 are conducted on other cancer cell lines and especially cancers in which the targeting of the avJ33 integrin is relevant such as melanoma, glioblastoma and other human ovarian cancer cell lines, particularly solid tumors.

Example 13: In Vivo Experiments with Compounds of Formula I and Complexes of Formula II

(59) In vivo therapeutic potential is evaluated by pre-clinical studies on cancer of mouse models and, in particular, cancers for which the RAFT(4RGD) group is a specific targeting moiety such as ovarian cancer, breast cancer, brain cancer, upper airways tract cancer, lung cancer, liver cancer, colon cancer, prostate cancer, bone cancer and their respective metastases.

(60) A/ In Vivo Biodistribution Study of Compounds of Formula I-1 and Complexes of Formula II-1.

(61) Compounds of formula I-1 and II-1 are administered intravenously or intraperitoneally to tumor-bearing animals (the above mentioned mouse models) and their biodistribution and pharmacokinetics are studied by non-invasive fluorescence imaging in the near infrared region using a Fluobeam700 apparatus (Fluoptics).

(62) From this study, the post-injection time at which the best signal-to-noise ratio is obtained is evaluated, corresponding to the photodynamic activation optimum of the compounds of formula I-1 and II-1 for a therapeutic use.

(63) B/ In Vivo Therapeutic Efficiency of the Compounds of Formula I-1 and II-1

(64) Compounds of formula I-1 and II-1 are administered intravenously or intraperitoneally to tumor-bearing animals (the above mentioned mouse models). At the post-injection time previously determined, the animals are submitted to fluorescence imaging (Fluobeam700; Fluoptics) to identify the tumors, non-invasively in the case of superficial tumors or intraoperatively for deeper tumors. The identified tumors are then irradiated by a near infrared laser (100-300 mW/cm.sup.2).

(65) In the non-invasive approach, the laser activation is optionally reproduced several times, and optionally preceded by a new injection of the compound.

(66) The therapeutic efficiency is evaluated by non-invasive diagnostic imaging measuring the tumor regression (bioluminescence, microCT, MRI, PET) and by the study of the survival curves of the animal models compared to untreated animals (control).

(67) In another attempt, the photodynamic treatment is done after a surgical tumor excision to get rid of the invisible tumor residues or tumor residues placed in a region where the surgical resection is impossible because of their close proximity to vital organs.