WO2013144405A1 - Novel chlorinated derivatives of bodipy - Google Patents

Description

 NEW CLOSED BODIPY DERIVATIVES

 SECTOR DESCRIPTION OF THE TECHNIQUE

The present invention relates in general to the field of dyes, to dyes with BODIPY structure in particular and specifically to chlorinated derivatives of BODIPY dyes.

STATE OF THE TECHNIQUE

Among the organic dyes, the 4, 4-difluoro-4-bora-3a, 4a-diaza-s-indacenes, commonly known under the trade name BODIPY, are considered today as one of the most useful and versatile dyes that exist. The interest in BODIPYs originates from its good photophysical properties, such as high absorption coefficients, quantum yields of fluorescence and photostability, as well as low sensitivity to the effects of the environment and high chemical versatility. At present, the development of new BODIPY fluorescent dyes has become a booming field of research, due to its possible applications as sensors in biology and in clinical diagnosis, as photosensitizers for photodynamic therapy (PDT), as systems laser, as waveguides, for the manufacture of light emitting diodes (OLED), as photovoltaic cells and as electroluminescent devices, in addition to their usual applications as organic dyes. These and other emerging applications are conditioned by the emission wavelength, the quantum performance and the stability of the dyes in the working conditions necessary for each specific use, which may be especially important in the development of new imaging techniques in microscopy. optics, which require a high intensity laser irradiation. In the latter case, one of the main problems related to the use of dyes, both as a source of laser radiation or as a biological marker, is its exposure to high pumping and temperature irradiations, which significantly limits its photostability, reducing the useful life of the dyes. Due to the wide applicability of these compounds, it is necessary to search for new BODIPY with stability and efficiency superior to those currently described. The introduction of different substituents and their distribution in the structure of BODIPYs is key to modulate and improve their properties. Several derivatives of BODIPY containing chlorine atoms bonded to the carbon atoms of the boradiazaindacene system are known. In most cases, the functionalization with this substituent is carried out with the intention of modulating its redox potential (Krumova, K _.; Cosa, GJ Am. Chem. Soc. 2010, 132, 17560-17569) and its pH values ( Baruah, M.; Qin, W .; Basaric, N.; De Borggraeve, WM; Boens, NJ Org. Chem. 2005, 70 (10), 4152-4157) or for use as a synthesis intermediate in the preparation of other compounds (Zhao, C; Zhang, Y .; Feng, P .; Cao, J. Dalton Trans. 2012, 41, 831-838. Ortiz, MJ; Agarrabeitía, AR; Duran-Sampedro, G.; Banuelos Prieto, J .; López, TA; Massad, WA; Montejano, HA; García, NA; López Arbeloa, I. Tetrahedron 2012, 68, 1153-1162. Read V. Leemans T. Boens N.; Dehaen, W. Eur. J . Org. Chem. 2011 4386-4396, Dodani, S. C.; Leary, S. C; Cobine, PA; Winge, DR; Chang, CJJ Am. Chem. Soc. 2011, 133, 8606-8616. Leen V Leemans T.; Boens N.; Dehaen, W. Eur. J. Org. Chem. 2011, 4386-4396. Leen, V .; Braeken, E .; Luckermans, K.; Jackers, C.; Van der Auweraer , M.; Boens,.; Dehaen, W. Chem. Commun. 2009, 4515-4517; Rohand, T. ; Baruah, M.; Qin. Boens, N.; Dehaen,. Chem. Commun. 2006, 266-268) or as a fluorescent marker (Kím, T.-I .; Park, S .; Choi, and. ; im, and. Chem. Asían J. 2011, 6, 1358-1361. Dilek, O .; Bañe, SLJ Fluoresc. 2011, 21, 347-354). The article in Chem Asían J. includes BODIPYs with chlorine in the R ² position (see formula I below) and its use as HOC1 sensors in cells. On the other hand, the article by J. Fluoresc. It includes BODIPYs with chlorine in the R ¹ position and its possible use to mark proteins, as is the article in J. Am. Chem. Soc. (2010) which includes BODIPYs with chlorine in R ² and R ^€ . However, none of these articles studies the photostability of the new halogenated dyes. All our compounds show high photostability and in this invention this property is correlated with the number and position of the chlorine atoms. None of the compounds described in these two articles fall within the definition of the compound of formula I of the present invention. The compounds described in these two documents contain amino and thioether groups bound directly to boradiazaindacene and are only useful sensors for substrates with aldehyde groups or for oxidizing species.

So far, the effect of introducing one or more chlorine atoms into the boradiazaindacene ring of BODIPY dyes and their use as a laser dye or fluorescent marker has not been studied in general. In the previous cases, obtaining these compounds, their properties and applications are limited by the presence of other functional groups and in no case is the presence of chlorine atoms directly correlated with an improvement in the optical properties and photostability of the dyes. . BRIEF DESCRIPTION OF THE INVENTION The present invention describes new dyes with a BODIPY structure characterized in that they comprise in its structure at least one chlorine atom covalently bonded to the boradiazaindacene carbon system of the dye. Most of the compounds of the present invention can be prepared by a new post-functionalization process, based on the reaction of boradiazaindacene with a chlorinating agent in the presence of an organic solvent.

This type of functionalization has been applied on different BODIPY dyes (in some commercial cases) commonly used as an active medium in lasers and with which emission at different wavelengths can be achieved. It has been observed that the presence of a single chlorine atom in the structure of the dye makes it possible to improve the photostability and laser efficiency of the dyes with respect to their unchlorinated analogs. This increase in optical properties is even greater as the number of chlorine atoms in the dye molecule increases.

These new compounds can be especially useful as an active medium in dye lasers both in the liquid phase and in the solid state, increasing their useful life and also in all those optical and analytical applications typical of the dyes such as sensors or markers in techniques of analysis based on laser spectroscopy. DETAILED DESCRIPTION OF THE INVENTION

donde , In a first aspect, the present invention describes the use of a compound of formula I

where ,

at least one of the substituents R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ is a chlorine atom attached directly to a carbon of the boradiazaindacene system, and

Substituents R ¹ , R ^~ , R ³ , R ⁴ , R ⁵ , R ^s and R 'other than chlorine are independently selected from the group consisting of hydrogen, halogen other than chlorine, cyano, alkyl, aryl, alkenyl, alkynyl, alkoxy and heterocycle, with the proviso that R ⁴ is not an alkyl substituted with hydroxyl or acyl;

as a laser dye or fluorescent marker.

The term "alkyl" refers, in the present invention, to radicals of hydrocarbon chains, linear or branched, having 1 to 10 carbon atoms, preferably 1 to 4, and which are attached to the rest of the molecule by a single bond, for example, methyl, ethyl, n-propyl, i-propyl, n-butyl, tere-butyl, sec-butyl, n-pentyl, Ώ-hexyl, etc. The alkyl groups may be substituted by one or more substituents such as halogen, hydroxyl, alkoxy, amino, carboxyl, acyl, alkoxycarbonyl, nitro, mercapto or alkylthio. The term "alkenyl" refers to hydrocarbon chain radicals of 2 to 25 carbon atoms, preferably 4 to 20, containing one or more double carbon-carbon bonds, for example, vinyl, 1-propenyl, allyl, isoprenyl, 2-butenyl, 1, 3 -butadienyl, hexenyl, octenyl, etc. Alkenyl radicals may be substituted by one or more substituents such as halogen, hydroxyl, alkoxy, amino, carboxyl, cyano, aryl, carbonyl, acyl, alkoxycarbonyl, amino, nitro, mercapto or alkyl.

The term "alkynyl" refers to hydrocarbon chain radicals of 2 to 25 carbon atoms, preferably 4 to 24, which contain one or more triple carbon-carbon bonds, for example, propynyl, butynyl, etc. Alkynyl radicals may be substituted by one or more substituents such as halogen, hydroxyl, alkoxy, carboxyl, cyano, aryl, amino, carbonyl, acyl, alkoxycarbonyl, amino, nitro, mercapto or alkylthio.

The term "acyl" refers, in the present invention, to radicals of linear or branched carboxylic acids, having 1 to 6 carbon atoms, and which are attached to the rest of the molecule by an ester bond. The acyl radicals may be substituted by one or more substituents such as halogen, hydroxyl, alkoxy, carboxyl, cyano, aryl, amino, carbonyl, acyl, alkoxycarbonyl, amino, nitro, mercapto or alkylthio.

The term "aryl" refers in the present invention to a monocyclic, bicyclic, tricyclic or tetracyclic aromatic hydrocarbon comprising an aromatic structure formed between 6 and 16 carbon atoms, for example, phenyl, naphthyl, indenyl, phenanthryl, anthracyl or pyrenyl. The aryl group may be substituted by one or more substituents such as alkyl, alkenyl, alkynyl, haloalkyl, amino, alkylamino, arylamino, dialkylamino, hydroxy, alkoxy, phenyl, mercapto, halogen, nitro, cyano, carbonyl or alkoxycarbonyl.

The term "heterocyclic" refers in the present invention to a stable monocyclic or polycyclic radical of 3 to 15 members, which may be unsaturated, saturated or partially saturated, and consisting of carbon atoms and at least one heteroatom selected from the group that It consists of nitrogen, oxygen and sulfur. Preferably it has 4 to 8 members with one or more heteroatoms and, more preferably, 5 to 6 members with one or more heteroatoms. Examples of heterocycles may be, not limited to: azepines, indols, imidazoles, isothiazoles, thiadiazoles, furans or thiophenes. For example, tetrahydrofuran, benzimidazole, benzothiazole, piperidine, piperazine, purine or quinoline. The term "alkoxy" refers to an R-0 group, where R is an alkyl, alkenyl, alkynyl or aryl as defined above.

In the present invention, "laser dye" is understood as a compound that can act as an active means to generate and / or emit laser radiation.

In the present invention, "fluorescent label" is understood as a compound comprising at least one chemical group, which after being excited with light is itself capable of emitting light. This fluorescent label can be attached to a second compound, preferably a biomolecule or a chemical, allowing the latter to be detected by techniques based on laser-induced fluorescence emission.

In a preferred embodiment, the present invention relates to the use of the compound of formula I as described in this patent application, wherein said compound comprises at least two chlorine atoms in the positions R ^~ , R ^¿ , R ³ , R ³ , R ° or R ', as a laser dye or fluorescent marker.

In an even more preferred embodiment, the present invention relates to the use of the compound of formula I as described in this patent application, wherein said compound comprises at least four chlorine atoms in the positions R ¹ , R ² , R ³ , R ⁵ , R ^s or R ⁷ , as a laser dye or fluorescent marker.

In another preferred embodiment, the compound of formula I is used as a fluorescent marker in a laser-induced fluorescence emission imaging and / or analysis technique, and said compound of formula I can be found covalently bound to a biomolecule selected from the group It consists of an amino acid, peptide, protein, lipid, carbohydrate, antibody, nucleic acid and toxin. In a more preferred embodiment the analysis technique may be laser microscopy.

In another preferred embodiment, the compound of formula I is used as a laser dye in a laser system used in an analytical, photoelectronic or biophotonic technique. The compounds of formula I of the present invention increase the efficiency and photostability of currently marketed laser systems.

In a second aspect, a fluorescent compound of formula I is described

 I

where

(a) R ⁴ is an aryl group,

R, R ³ , R ⁵ and R ⁷ are independently selected from the group consisting of hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl and heterocycle,

at least one of the substituents R ² or R ^s is a chlorine atom and the other substituent, if not chlorine, is independently selected from the group consisting of hydrogen, halogen other than chlorine, alkyl, aryl and heterocycle; or

(b) R ⁴ is an unsubstituted alkyl group,

R ¹ , R ³ , R ⁵ and R 'are independently selected from the group consisting of halogen, alkyl, alkenyl, alkynyl, aryl and heterocycle,

at least one of the substituents R ² or R ⁶ is a chlorine atom and the other substituent, if it is not chlorine, is independently selected from the group consisting of hydrogen, halogen other than chlorine, alkyl, aryl and heterocycle; or

(c) R ⁴ , R °, R ^s and R ⁷ are the same or different alkyl groups, at least one of the substituents R ¹ , R ² or R ³ is a chlorine atom and the other substituents, if not chlorine , are independently selected from the group consisting of hydrogen, alkyl, aryl and heterocycle. In a preferred embodiment, the present invention relates to a compound of formula I as defined in (a) where R ⁴ may be a monocyclic aromatic hydrocarbon, which may be substituted by one or more substituents such as alkyl, alkenyl, alkynyl or aryl. More preferably R ⁴ is phenyl or phenyl substituted with a C1-C4 alkyl. Especially preferred, R ⁴ is tolyl.

Preferably, the invention as described in this patent application refers to a compound of formula I as defined in (a) where R ¹ , R ³ , R ^s and R ⁷ are independently selected from the group consisting in hydrogen, halogen and monocyclic aryl, it being especially preferable that R ⁴ is also a tolyl group.

In an even more preferred embodiment, the compound of formula I of the present invention as defined in (a), preferably when R ⁴ is tolyl and R ¹ , R ³ , R ⁵ and R ⁷ are independently selected from the group consisting of hydrogen, halogen and monocyclic aryl, it is selected from the group consisting of 2, 3, 5, 6, 7, 9 and 10.

 10

In another preferred embodiment, the present invention relates to a compound of formula I as defined in (a), preferably when R ⁴ is tolyl, where R ¹ , R ² , R ^s and R ⁷ are chlorine atoms.

In another preferred embodiment, the present invention relates to a compound of formula I as defined in (a) where R ¹ , R ² , R ³ , R ⁵ , R ^s and R ⁷ are chlorine atoms. Preferably, R ⁴ may be a monocyclic aromatic hydrocarbon, which may be substituted by one or more substituents such as alkyl, alkenyl, alkynyl or aryl.

In another preferred embodiment, the present invention relates to a compound of formula I as defined in (b), where preferably R ¹ , R ³ , R ⁵ and R ⁷ are independently selected from the group consisting of hydrogen, halogen or alkyl. More preferably R ⁴ is a C1-C10 alkyl, and R ¹ , R ³ , R ⁵ and R ⁷ are independently selected from hydrogen, halogen or C1-C4 alkyl. Even more preferably, R ⁴ is a C1-C10 alkyl and R ¹ , R ³ , R ⁵ and R ⁷ are methyl.

Preferably, the compound of formula I as defined in (b) of the present invention, where R ⁴ is a C 1 -C 10 alkyl and R ¹ , R ³ , R ³ and R ⁷ are methyl groups, is selected from the group that It comprises 16 and 17.

 16 17

In another preferred embodiment, the invention described in this patent application refers to a compound of formula I as defined in (c), where R ⁴ , R ⁵ , R ^s and R ⁷ are equal C1-C4 alkyl groups or different, R ¹ is chlorine, and R ² and R ³ are independently selected from the group consisting of hydrogen and chlorine. Preferably, the compound of formula I of the present invention is selected from the group consisting of 12, 13 and 14.

En otra realización preferida, la presente invención también se refiere al uso como colorante láser o marcador fluorescente del compuesto fluorescente de fórmula I tal como se describe en este segundo aspecto de la invención. Preferiblemente, cuando el compuesto se utiliza como marcador fluorescente en una técnica de imagen y/o análisis basada en emisión de fluorescencia inducida por láser, y dicho compuesto de fórmula I se puede encontrar unido covalentemente a una biomolécula seleccionada del grupo que consiste en un aminoácido, péptido, proteína, lípido, carbohidrato, anticuerpo, ácido nucleico y toxina. En una realización mas preferida la técnica de análisis puede ser la microscopía láser. Por otro lado, también se prefiere que el compuesto de fórmula I se utilice como colorante láser en un sistema láser utilizado en una técnica analítica, fotoelectrónica o biofotónica. Los compuestos de fórmula I de la presente invención aumentan la eficacia y fotoestabilidad de los sistemas láseres actualmente comercializados.

In another preferred embodiment, the present invention also relates to the use as a laser dye or fluorescent marker of the fluorescent compound of formula I as described in this second aspect of the invention. Preferably, when the compound is used as a fluorescent marker in an image and / or analysis technique based on laser induced fluorescence emission, and said compound of formula I can be found covalently bound to a biomolecule selected from the group consisting of an amino acid , peptide, protein, lipid, carbohydrate, antibody, nucleic acid and toxin. In a more preferred embodiment the analysis technique may be laser microscopy. On the other hand, it is also preferred that the compound of formula I be used as a laser dye in a laser system used in an analytical, photoelectronic or biophotonic technique. The compounds of formula I of the present invention increase the efficiency and photostability of currently marketed laser systems.

In a third aspect of the invention a method of obtaining a compound of formula I is described as described in this patent application in options (a) or (b), both in its more general definition and in preferred embodiments. described in this patent application, characterized in that it comprises a chlorination reaction between a compound of formula II and a chlorination agent in the presence of an organic solvent,

donde al menos un sustítuyente R^2' o R^s' es un átomo de hidrógeno unido directamente al sistema boradiazaindaceno y el otro sustítuyente, si no es hidrógeno, se selecciona de forma independíente del grupo que consiste en halógeno, alquilo, arilo y heterociclo,

where at least one R ^{2 '} or R ^s' substituent is a hydrogen atom attached directly to the boradiazaindacene system and the other substituent, if not hydrogen, is independently selected from the group consisting of halogen, alkyl, aryl and heterocycle,

R ^' , R ^3' , R ^{5 '} and R ^7' are independently selected from the group consisting of hydrogen, halogen, alkenyl, alkynyl, aryl and heterocycle, and

R ^{4 '} is an aryl or unsubstituted alkyl group.

In a preferred embodiment, the present invention relates to a process for obtaining a compound of formula I as defined in options (a) or (b), both in its more general definition and in the preferred embodiments described in this patent application, which comprises the chlorination of a compound of formula II where the substituents R ^{1 '} , R ^3' , R ⁴ , R ^{5 '} and R ^7' have the same meaning as the corresponding substituents R ¹ , R ³ , R ⁴ , R ⁵ and R ⁷ in the compound of formula I defined in this patent application.

In an even more preferred embodiment, the present invention relates to a process for obtaining a compound of formula I as defined in the preceding paragraphs, which comprises reacting a compound of formula II where R ^{2 '} and R ^6' are hydrogens, with a chlorination agent in the presence of an organic solvent to obtain a compound of formula I where R ² and R ^s are chlorine. A chlorinating agent is understood as a reagent capable of inducing the formation of a chloro-carbon covalent bond in an aromatic or aliphatic system.

The process for obtaining a compound of formula I as described in this patent application may comprise the use of a chlorination agent selected from the group consisting of chlorine, a salt of hydrochlorite, phosphorus pentachloride, a transition metal chloride, or an organic compound containing one or more chloro-amino bonds such as N-chlorosuccinimide or N, N '-dichlorobis- (2, 4, 6- trichlorophenyl} urea Preferably, the chlorinating agent is N-chlorosuccinimide (NCS).

The process for obtaining a compound of formula I as described in this patent application can take place in the presence of an organic solvent that is selected from the group consisting of water, acetonitrile, N, N-dimethylformamide, N, iV- dimethylmettamide, dichloromethane, 1,2-dichloroethane, chloroform, carbon tetrachloride, diethyl ether, t-butyl methyl ether, tetrahydrofuran, 1,4-dioxane, ethyl acetate, isopropyl acetate, methanol, ethanol, 1-propanol, 2 -propanol, 1-butane1 or t-butanol. Preferably, the organic solvent is tetrahydrofuran (THF). In a particularly preferred embodiment, the process for obtaining a compound of formula I as described in this patent application comprises the use of AT-chlorosuccinimide as a chlorinating agent and tetrahydrofuran (THF) as an organic solvent.

The process for obtaining a compound of formula I as described in this patent application can take place at a temperature between 0 and 140 ° C. Preferably, the reaction takes place at a temperature between 10 and 70 ° C.

Throughout the description and the claims the word "comprises" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and features of the invention will be derived partly from the description and partly from the practice of the invention. The following examples and drawings are provided by way of illustration, and are not intended to be limiting of the present invention.

EXAMPLES

TO . SYNES IS OF THE COMPOUNDS OF THE INVENTION

 12 CI H H

_R i R ² R ⁵ R ⁶ R ⁷

 13 CI H Cí

1 H H H H H H 14 Cf CI H

5 Cf CI HHH CI R ² R ⁶

6 Ci Ci H H Cí CI 15 H H

 16 H C¡

7 Ct CI is Cí Cí CI

17 Cí Cí β 4-C ₃ H ₄ CHO HHHH 4-C ₃ H ₄ CHO

θ 4-CeH ₄ CHO Cí HHH 4-C ₆ HCHO

10 4-C _and H ₄ CHO CI HH Cí 4-C ₅ H ₄ CHO

All reagents used in the preparation of BODIPYs are commercial or prepared according to methods previously described in the literature. The solvents were dried and distilled before use. The spectroscopic data of the starting compounds was found to match the data in the literature. The products were purified by flash column chromatography using Merck 60 silica gel (230-400 mesh). The ^X H and ¹³ C NMR spectra were performed on a Bruker Avance DPX-300 spectrometer (300 MHz for ^X H and 75 MHz for ¹³ C) and a Bruker Avance III spectrometer (700 MHz for ¹ H and 176 MHz for ¹³ C) . All spectra were performed in CDC1 ₃ . The ¹ H chemical shifts are indicated in ppm relative to tetramethylsinola (δ = 0.00 ppm), using the residual solvent signal as internal reference. Chemical shifts of ^x3 C are indicated in ppm using CDC1 ₃ (δ = 77.67 ppm) as internal standard. The multiplicities in the chemical shift are indicated as s = singlet, d = doublet, t = triplet, q = quartet and m multiplet. IR spectra (in cm ^~~ ) were performed on a Bruker Tensor-27-FTIR spectrometer. Melting points were determined in open capillaries and are uncorrected. The high resolution mass spectra were performed by ESI in an FTMS Bruker APEX Q IV and by El in a Thermofísher MAT 95 XP.

BODIPY dyes 1, 4, 8 and 11 were synthesized according to methods described earlier in the literature

(Cuí, A .; Peng, X .; Fan, J .; Chen, X .; u, Y .; Guo, B. J.

Photochem Photobiol A 2007, 186, 85-92; Rohand, T. ;

Baruah, M .; Qin, W .; Boens, N.; Dehaen, W. Chem. Commun.

2006, 266-268; Ortíz, M. J .; García-Moreno, I .; Agarrabeítia, A. R.; Duran-Sampedro, G.; Costela, A.;

Tailor, R.; López Arbeloa, F. ; Bañuelos Prieto, J.; Lopez

Arbeloa I. Phys. Chem. Chem. Phys. 2010, 12, 7804-7811;

Bañuelos-Prieto, J .; Agarrabeítia, A. R .; Garcia-Moreno,

I .; López -Arbeloa, I .; Costela, A .; Infantes, L .; Perez- Ojeda, M. E. ; Palaces-Cuesta, M. ; Ortiz, M. J. Chem. Eur.

J. 2010, 16, 14094-14105). BODIPY 15 dye was purchased from Lasing, S. A.

General procedure for chlorination of BODIPYs

On a stirring solution of BODIPY in anhydrous THF N-chlorosuccinimide (NCS) in anhydrous THF is added to room temperature and the mixture is stirred for 12-72 h under argon atmosphere. At the end of the reaction, the mixture is diluted with AcOEt, washed with 10% HC1 and H ₂ 0, dried with MgSO ₄ and evaporated to dryness. Chlorinated dyes are purified by column chromatography.

2-Chloro-4,4-difluoro-8- (4-tolyl) -4-bora-3a, 4a-diaza-s-indacene (2): following the general chlorination procedure, BODIPY 1 (50 mg, 0.18 mmol) in anhydrous THF (20 mL) with NCS (47 mg, 0.36 mmol) in anhydrous THF (5 mL) for 12 h. Purification was performed by column chromatography using hexane / EtOAc (98: 2) and 2 (45 mg, 80%) was obtained as a green-orange solid, mp 171.8-172.0 ° C; ¹ H NMR (700 MHz, CDC1 ₃ ): δ 7.92 (1 H, s, H-5), 7.68 (1 H, s, H-3), 7.39 (2 H, d, J = 8.4 Hz, H- 2 ^" ), 7.28 (2 H, d, J = 8.4 Hz, H-3 ^' ), 6.95 (1 H, d, J = 4.2 Hz, H-7), 6.74 (1 H, s, Hl), 6.52 (1 H, d, J = 4.2 Hz, H-6), 2.41 (3 H, s, CH ₃ ); ¹³ C NMR (176 MHz, CDCI3): δ 147.8 (C-8), 145.8 (CH-5 ), 142.0 (C-4 ^' ), 139.7 (CH-3), 135.4 (C-7a), 133.2 (C-8a), 133.1 (CH-7), 130.6 (2 CH-2 ^" ), 130.5 (Cl ^" ), 129.4 (2 CH-3 ^' ), 127.3 (CH-1), 121.5 (C-Cl), 119.3 (CH-6), 21.5 (CH ₃ ); IR (pure): 1548, 1405, 1364, 1259, 1105, 985 crn ^"1 ; HRMS-ESI *: calculated (Ci ₆ H ₁₂ BClF ₂ N ₂ + Na ⁺ ) 339.0648 found 339.0628.

2, 6-Dichloro- 4, 4-difluoro-8- (4-tolyl) -4-bora-3a, 4a-diaza-s-indacene (3): following the general chlorination procedure, BODIPY 1 was reacted (40 mg, 0.14 mmol) in anhydrous THF (15 mL) with NCS (56 mg, 0.42 mmol) in anhydrous THF (5 mL) for 24 h. Purification was performed by column chromatography using hexane / EtOAc (98: 2) and 3 (39 mg, 85%) was obtained as an orange solid. mp 196.3-197.0 ° C; ¹ H NMR (700 MHz, CDCI ₃ ): δ 7.73 (2 H, s, H-3 and H-5), 7.37 (2 H, d, J = 8.4 Hz, H-2 ^" ), 7.29 (2 H , d, J = 8.4 Hz, H-3 ^" ), 6.79 (2 H, s, Hl and H-7), 2.41 (3 H, s, CH ₃ ); ¹³ C NMR (176 MHz, CDCI ₃ ): δ 148.0 (C-8), 142.5 (C-4 ^' ), 141.8 and 141.7 (CH-3 and CH-5), 133.6 (C-7a and C-8a), 130.6 (2 CH-2 '), 130.1 (C-l'), 129.6 (2 CH-3 '}, 128.6 and 128.5 (CH-1 and CH-7), 122.6 (C2-C1 and C6-Cl), 21.6 (CH ₃ ); IR (pure): 1550, 1484, 1358, 1261, 1112 cm ^"1 ; HRMS-ESI: calculated for {Ci ₆ HiiBCl ₂ F2 2+ a ⁺ ) 373.0258 found 373.0187.

2,3,5-Trichloro-4, -difluoro-8- (4- tolyl) -4-bora-3a, 4a-diaza-s-indacene (5): following the general chlorination procedure, the BODIPY was reacted 1 (90 mg, 0.32 mmol) in anhydrous THF (15 mL) with NCS (171 mg, 1.28 mmol) in anhydrous THF (10 mL) for 72 h. Purification was performed by column chromatography using hexane / EtOAc (98: 2) and 5 (40 mg, 32%) was obtained as an orange-red solid, together with an inseparable mixture of chlorinated products, mp 194.3-195.0 ° C ; ¹ H NMR (700 MHz, CDC1 ₃ ): δ 7.31 (2 H, d, J = 8.4 Hz, H-2 '}, 7.27 (2 H, d, J = 8.4 Hz, H-3'), 6.86 ( 1 H, d, J = 4.2 Hz, H-7), 6.74 (1 H, s, Hl), 6.41 (1 H, d, J = 4.2 Hz, H-6), 2.41 (3 H, s, CH ₃ ); ¹³ C NMR (176 MHz, CDC1 ₃ ): δ 146.6 (C-8), 144.3 (C5-C1), 142.0 (C-4 '), 140.6 (C3-C1)), 134.1 (C-7a ), 132.8 (CH-7), 131.1 (C-8a), 130.5 (2 CH-2 "), 129.5 (2 CH-3 ^' ), 129.2 (Cl"), 127.2 (CH-1), 120.6 (C2 -C1), 119.6 (CH-6), 21.5 (CH ₃ ); IR (pure): 1554, 1387, 1258, 1107, 800 cm ¹ ; HRMS-ESI ⁺ : calculated for (Ci ₆ H ₁₀ BCl ₃ F ₂ N ₂ + Na) 406.9865 found 406.9834.

2,3,5,6-Tetrachloro-, 4-difluoro-8- (4- tolyl) -4-bora-3a, 4a-diaza-s-indacene (6): following the general chlorination procedure, reacted BODIPY 1 (65 mg, 0.23 mmol) in anhydrous THF (20 mL) with NCS (308 mg, 2.3 mmol) in anhydrous THF (10 mL) for 48 h. Purification was performed by column chromatography using hexane / EtOAc (98: 2) and 6 (75 mg, 78%) was obtained as an orange-red solid. mp 231.0-231.8 ° C; ¹ H NMR (700 MHz, CDC1 ₃ ): δ 7.30 (2 H, d, J = 8.4 Hz, H-2 '), 7.28 (2 H, d, J = 8.4 Hz, H-3 "), 6.79 ( 2 H, s, Hl and H-7), 2.41 (3 H, s, CH ₃ ); ¹³ C NMR (176 MHz, CDC1 ₃ ): δ 144.4 (C-8), 142.6 (C3-C1 and C5-C1), 142.4 (C-4 "), 131.4 (C-7a and C-8a), 130.5 (2 CH-2"), 129.6 (2 CH-3 "), 128.9 (Cl"), 128.2 (CH-1 and CH-7), 121.6 (C2-C1 and C6-C1), 21.5 (CH ₃ ); IR (pure): 1549, 1384, 1245, 1106 was ^"1 ; HRMS-EI: calculated for (Ci ₆ H ₃ BCl ₄ F _{2 2} ) 417.9579 found 417.9583.

1,2,3, 5,6, 7-Hexachloro-4,4-difl gold-8- (4-tolyl) -4-bora-

3a, 4a-diaza-s-indacene (7): 5- (4- tolyl) dipyrromethane 18 (Littler, BJ; Miller, MA; Hung, C.-H .; Wagner, RW; O'Shea was reacted , DF; Boyle, P. D .; Lindsey, JSJ Org. Chem. 1999, 64, 1391-1396) (400 mg, 1.69 mmol) in anhydrous THF (30 mL) with a solution of NCS (3.38 mg, 23.5 mmol ) in anhydrous THF (15 mL) for 24 h under reflux under argon. The solvent was evaporated and DDQ (422 mg, 1.86 mmol) was added to the solution in CH ₂ C1 ₂ (50 mL) of the previously generated hexachlorodipyrromethane. The mixture was stirred at room temperature for 1 h and triethylamine (1.5 mL, 8.4 mmol) was added. After 10 minutes, BF ₃ -Et ₂ 0 (2 mL, 13.5 mmol) was added and the solution was stirred 24 h at room temperature. The reaction mixture was washed with H ₂ 0, dried over MgSO ₄ , filtered and evaporated to dryness. Purification was performed by column chromatography using hexane / EtOAc (99: 1) and 7 (245 mg, 30%) was obtained as a garnet solid, mp 275.1-276.0 ° C; Κ NMR (700 MHz, CDCl ₃ ): δ 7.26 (2 H, d, J = 8.4 Hz, H-2 "), 7.06 (2 H, d, J ^" = 8.4 Hz, H-3 "), 2.40 ( 3 H, s, CH ₃ ); ¹³ C NMR (176 MHz, CDC1 ₃ ): δ 143.7 (C-8), 142.5 (C-4 ") 140.7 (C3-C1 and C5-C1), 132.9 (C- 7a and C-8a), 130.1 (2 CH-2 "), 127.7 (2 CH-3"), 127.6 (C-l '), 126.3 (Cl-Cl and C7-Cl), 121.4 (C2-C1 and C6-C1), 21.6 (CH ₃ ); IR (pure): 1552, 1383, 1246, 1108 was ^"1 ; HRMS-ESI ^" : calculated for (C ₁₆ H ₇ BCl ₆ F ₂ N ₂ + CH ₃ OH-H) 518.8964 found 518.8926.

2-Chloro-4,4-difluoro-3,5-bis (4-formylphenyl) -8- (4-tolyl) -4- bora-3a, 4a-diaza-s-indacene (9): following the general procedure of chlorination, BODIPY 8 (80 mg, 0.16 mmol) was reacted in anhydrous THF (15 mL) with NCS (43 mg, 0.32 rnmol) in anhydrous THF (10 mL) for 48 h. Purification was performed by column chromatography using hexane / EtOAc (9: 1) and 9 (55 mg, 65%) was obtained as a red solid. mp 182.4-182.9 ° C; ¾ NMR (700 MHz, CDCl ₃ ): δ .9.99 (1 H, s, CHO), 9.95 (1 H, s, CHO), 7.91 (2 H, d, J = 8.4 Hz, 4-formylphenyl), 7.90 (2 H, d, J = 8.4 Hz, 4- formylphenyl), 7.83 (2 H, d, J = 8.4 Hz, 4-formylphenyl), 7.79 (2 H, d, J = 8.4 Hz, 4-formylphenyl), 7.44 (2 H, d, J = 8.4 Hz, 4-tolyl), 7.32 (2 H, d, J = 8.4 Hz, 4-tolyl), 6.99 (1 H, d, J = 4.2 Hz, H-7) , 6.87 (1 H, s, Hl), 6.66 (1 H, d, J = 4.2 Hz, H-6), 2.44 (3 H, s, CH ₃ ); ¹³ C NMR (176 MHz, CDC1 ₃ ): δ 191.9 (CHO), 191.7 (CHO), 158.9 (C), 151.3 (C), 146.2 (C), 141.8 (C), 137.5 (C), 137.4 (2 C), 136.7 (C), 136.6 (C), 135.5 (C), 133.1 (CH), 131.1 (2 CH), 131.0 (C), 130.7 (2 CH), 130.0 (2 CH), 129.6 (2 CH ), 129.4 (2 CH), 129.2 (2 CH), 127.8 (CH), 122.1 (C-Cl), 121.9 (CH); IR (pure): 2926, 2860, 1702, 1542, 1260, 1073, 800 cm ^"1 ; HRMS-EI: calculated for (C ₃ 0H ₂ 0BCIF _{2 2} O ₂ ) 524.1272 found 524.1280.

2, 6-Dichloro-4,4-difluoro-3, 5-bis (4- formylphenyl) -8- (4- tolyl) -4-bora-3a, 4a-diaza-s-indacene (10): following the general chlorination procedure, BODIPY 8 (80 mg, 0.16 mmol) was reacted in anhydrous THF (15 mL) with NCS (85 mg, 0.64 mmol) in anhydrous THF (10 mL) for 3 h at reflux. Purification was performed by column chromatography using hexane / EtOAc (9: 1) and 10 (55 mg, 62%) was obtained as a red solid. mp 211.3-211.8 ° C; ¾ NMR (700 MHz, CDCI ₃ ): δ 9.98 (2 H, s, 2 CHO), 7.87 (4 H, d, J = 7.7 Hz, H-3 "), 7.75 (4 H, d, J = 7.7 Hz, H-2 ^" ), 7.44 (2 H, d, J = 8.4 Hz, H-2 ^"" ), 7.34 (2 H, d, J = 8.4 Hz, H-3 ^"" ), 6.93 (2 H , s, Hl and H-7), 2.45 (3 H, s, CH ₃ ); ¹³ C NMR (176 MHz, CDCI ₃ ): δ 191.8 (2 CHO), 153.2 (C-3 and C-5), 146.4 (C- 8), 142.2 (C-4 "), 136.8 (C-4" ), 134.9 (Cl ^" ), 133.4 (C-7a and C-8a), 130.9 (4 CH-2"), 130.6 (2 CH-2 "^" ), 130.3 (Cl ^'" ), 129.6 (2 CH- 3 "), 129.2 (4 CH-3 ^" ), 129.1 (CH-1 and CH-7), 123.0 (C2-C1 and C6-C1), 21.6 (CH ₃ ); IR (pure): 2924, 2856, 1700, 1540, 1268, 1219, 1144, 1070, 800, 768 ctlf ¹ ; HRMS-ESI ^" : calculated for (C _3G H _I9 BC1 ₂ F ₂ N ₂ 0 ₂ + CH ₃ OH-H ⁺ ) 589.1169 found 589.1234. 2-Chloro-4, 4-difluoro-1, 3,5,7, 8-pentamethyl-4-bora-3a, 4-diaza-s-indacene (16): following the general chlorination procedure, BODIPY 15 (60 mg, 0.23 mmol) was reacted in anhydrous THF (15 mL) with NCS (85 mg, 0.64 mmol) in anhydrous THF (5 mL) for 48 h. Purification was performed by column chromatography using hexane / CHCl ₃ (7: 3) and 16 (50 mg, 73%) was obtained as a solid. orange. mp 259.4-259.7 ° C; ¹ H NMR (700 MHz, CDC1 ₃ ): δ 6.03 (1 H, s, H-6), 2.53 (3 H, s, CH ₃ -C8), 2.46 (6 H , s, CH ₃ -C3 and CH ₃ -C5), 2.36 (3 H, s, CH ₃ -C7), 2.34 (3 H, s, CH ₃ -CI); ¹³ C NMR (176 MHz, CDC1 ₃ ) : δ 156.0 (C-5), 148.3 (C-3), 142.8 (C-7), 141.7 (C-8), 134.1 (Cl), 132.7 (C-7a), 129.8 (C-8a), 122.1 (CH-6), 121.3 (C-Cl), 17.5 (CH ₃ -C7), 16.7 (CH ₃ -C8), 14.6 (CH ₃ -C5), 14.4 (CH ₃ -C1), 12.2 (CH ₃ - C3); IR (pure): 1560, 1365, 1268, 1110 cnf ¹ ; HRMS-ESI ^"1" : calculated for (Ci ₄ H ₁₆ BClF ₂ N ₂ + Na ⁺ ) 319.0961 found 319.0980.

2, 6-Dichloro-4, 4-difluoro-1, 3, 5,7, 8-pentamethyl-4-bora-3a, 4a-diaza-s-indacene (17): following the general chlorination procedure, it was made react the BODIPY 15 (50 mg, 0.19 mmol) in anhydrous THF (15 mL) with NCS (76 mg, 0.57 mmol) in anhydrous THF (10 mL) for 24 h. Purification was performed by column chromatography using hexane / EtOAc (98: 2) and 17 (42 mg, 67%) was obtained as an orange solid. mp 261.1-261.6 ° C; Hi NMR (700 MHz, CDCI ₃ ): δ 2.56 (3 H, S, CH ₃ -C8), 2.48 (6 H, s, CH ₃ -C3 and CH ₃ - C5), 2.35 (3 H, s, CH ₃ -C1 and CH ₃ -C7); ¹³ C NMR (176 MHz, CDCI ₃ ): δ 150.7 (C-3 and C-5), 142.2 (C-8), 135.8 (Cl and C- 7), 130.4 (C-7a and C-8a), 122.4 (2 C-Cl), 17.1 (CH ₃ -C8), 14.6 (CH ₃ -CI and CH ₃ -C7), 12.3 (CH ₃ -C3 and CH ₃ -C5); IR (pure): 1560, 1362, 1270, 1111 cm ^"1 ; HRMS-EI: calculated for (C ₁₄ H ₁₅ BC1 ₂ F ₂ N ₂ ) 330.0668 found 330.0668. General procedure for the synthesis of BODIPYs 12-14

To a solution of chlorinated 2-acetylpyrrole (1 equiv.) In CHC1 ₃ POCl ₃ (1.1 equiv.) Was added and the mixture was stirred for 30 min at room temperature. 3-Ethyl-2,4-dimet-ilpirrol (1 equiv.) In CHC1 _{3 was added} and the resulting solution was stirred for 12 h at room temperature. Triethylamine (1 equiv.) Was added, followed by addition of BF ₃ · Et ₂ 0 (1 equiv.), And stirring was continued for 3.4 h, was added aqueous 10% HC1 and extraction was performed with CH ₂ C1 ₂ . The organic phase was washed with water, dried over MgSO ₄ , filtered and evaporated to dryness. The dyes were isolated and purified by flash chromatography on silica gel.

Chlorination of acetylpyrrole

On a solution of 2-acetylpyrrole (200 mg, 1.83 mmol) in anhydrous THF (30 mL) NCS (269 mg, 2.01 mmol) in anhydrous THF (30 mL) was added and the mixture was stirred at room temperature for 24 h . Purification was performed by column chromatography using hexane / EtOAc (95: 5) and 2-acetyl-5-chloropyrrole 19 (Ho, C.-T .; Jín, QZ; Lee, K. N .; Carlin , JTJ Agrie. Food Chem. 1982, 30, 362-364. B) Leen, V .; Braeken, E .; Luckermans, K.; Jackers, C.; Van der Auweraer, M; Boens, N.; Dehaen, W. Chem. Commum. 2009, 4515-4517. c) Rane, R. A .; Telvekar, V. N. Bioorg. Med. Chem. Lett. 2010, 20, 5681-5685) (110 mg, 42%) as a colorless solid.

On a solution of 2-acetylpyrrole (200 mg, 1.83 mmol) in anhydrous THF (30 mL) NCS (734 mg, 5.5 mmol) in anhydrous THF (50 mL) was added and the mixture was stirred at room temperature for 72 h. Purification was performed by column chromatography using hexane / EtOAc (95: 5) and 2-acetyl-3, 5-dichloropyrrole (20) (135 mg, 42%) was obtained. as a colorless solid and 5-acetyl-2,3-dichloropyrrole (21) (135 mg, 42%) as a colorless solid.

2 - cetyl -3, 5-dichloropyrrole (20): ^X H NMR (300 MHz, CDC1 ₃ ): δ 10.09 (1 H, wide singlet, H), 6.07 (1 H, d, J = 3.0 Hz, H- 3), 2.51 (3 H, s, CH ₃ ); ¹³ C NMR (75 MHz, CDCl ₃ ): δ 186.7 (CO), 127.6 (C-2), 122.5 (C5-C1), 119.7 (C3-C1), 110.4 (CH), 28.1 (CH ₃ ); IR (pure): 3218, 1643, 1446, 1400, 761 cnf ¹ ; HRMS-EI: calculated for (C ₆ H _S C1 ₂ N0) 176.9749 found 176.9751.

5-Acetyl-2, 3-dichloropyrrole (21): ^X H NMR (300 MHz, CDC1 ₃ ): δ 10.81 (1 H, wide singlet, NH), 6.76 (1 H, d, J = 2.7 Hz, H- 4), 2.51 (3 H, s, CH ₃ ); ¹³ C NMR (75 MHz, CDCl ₃ ): δ 186.3 (CO), 128.0 (C-5), 120.5 (C2-C1), 115.5 (CH), 110.5 (C3-Cl), 23.9 (CH ₃ ); IR (pure): 3218, 1643, 1447, 1399, 768 cm ^{~ x} ; HRMS-EI: calculated for (C _s H ₅ Cl ₂ NO) 176.9749 found 176.9750. 5-Chloro-2-ethyl-4, 4-difluoro-1, 3, 8-trimethyl-4-bora-3a, 4a-diaza-s-indacene (12): 2-Acetyl-5-chloropyrrole was reacted (19) (91 mg, 0.63 mmol) in CHC1 ₃ (10 mL), with P0C1 ₃ (0.06 mL, 0.7 mmol), 3 -etl-2, 4-dimet-ilpirrol (82 mg, 0.63 mmol) in CHC1 ₃ (10 mL), triethylamine (0.08 mL, 0.63 mmol) and BF ₃ -Et ₂ 0 (0.08 mL, 0.63 mmol). Purification was performed by column chromatography using hexane / CHCl ₃ (5: 5) and 12 (98 mg, 52%) was obtained as an orange solid, mp 171.2-171.7 ° C; E NMR (700 MHz, CDCl ₃ ): δ 6.90 (1 H, d, J = 4.2 Hz, H-7), 6.20 (1 H, d, J = 4.2 Hz, H-6), 2.51 (3 H, s, CH ₃ ), 2.44 (3 H, s, CH ₃ ), 2.35 (2 H, q, J = 7.7 Hz, CH ₂ ), 2.26 (3 H, s, CH ₃ ), 0.99 (3 H, t , J = 7.7 Hz, CH ₃ CH ₂ ); ¹³ C NMR (176 MHz, CDCl ₃ ): δ 161.1 (C), 140.7 (C), 138.6 (C), 135.7 (2 C), 134.0 (C), 133.1 (C), 122.8 (CH), 114.5 ( CH), 17.1 (CH ₂ ), 15.9 (CH ₃ ), 14.5 (CH ₃ ), 14.1 (CH ₃ ), 13.1 (CH ₃ ); IR (pure): 1577, 1403, 1212, 1101, 771 cm ^"1 ; HRMS-EI: calculated for (C ₁₄ H ₁₆ BC1F ₂ N ₂ ) 296.1060 found 296.1064. 5, 7-Dichloro-2-ethyl-4,4-difluoro-1, 3, 8-trimethyl-4-bora-3a, a-diaza-s-indacene (13): 2- Acetyl-3 was reacted , 5-dichloropyrrole (20) (90 mg, 0.50 mmol) in CHC1 ₃ (10 mL), with POCl ₃ (0.05 mL, 0.55 mmol), 3-ethyl-2,4-dimethylpyrrole (91 mg, 0.50 mmol) in CHC1 ₃ (10 mL), triethylamine (0.06 mL, 0.50 mmol) and BF ₃ -Et ₂ 0 (0.06 mL, 0.50 mmol). Purification was performed by column chromatography using hexane / EtOAc (98: 2) and 13 (90 mg, 54%) was obtained as an orange solid, mp 193.1-193.6 ° C; ¹ H NMR (700 MHz, CDC1 ₃ ): δ 6.21 (1 H, s, H-6), 2.72 (3 H, s, CH ₃ ), 2.51 (3 H, s, CH ₃ ), 2.36 (2 H , q, J = 7.7 Hz, CH ₂ ), 2.31 (3 H, s, CH ₃ ), 1.00 (3 H, t, J = 7.7 Hz, CH ₃ CH ₂ ); ¹³ C NMR (176 MHz, CDC1 ₃ ): δ 162.2 (C), 141.7 (C), 139.8 (C), 137.0 (2 C), 135.0 (C), 133.2 (C), 127.2 (C), 126.4 ( C), 115.6 (CH), 17.2 (CH ₂ ), 16.3 (CH ₃ ), 14.9 (CH ₃ ), 14.4 (CH ₃ ), 13.2 (CH ₃ ); IR (pure): 1568, 1417, 1375, 1194, 1084, 1031, 803 cm ^'1 ; HRMS-EI: calculated for (Ci ₄ Hi ₅ BCl ₂ F ₂ N ₂ ) 330.0668 found 330.0669.

5, 6-Dichloro-2-ethyl-4, 4-difluoro-1, 3, 8-trimethyl-4-bora-3a, 4a-diaza-s-indacene (14): 2,3-Dichloro was reacted -5-acetylpyrrole (21) (90 mg, 0.50 mmol) in CHC1 ₃ (10 mL) with _POCl3 (0.05 mL, 0.55 mmol), 3-ethyl-2,4- dimethylpyrrole (91 mg, 0.50 mmol) in CHC1 ₃ (10 mL), triethylamine (0.06 mL, 0.50 mmol) and BF ₃ -Et ₂ 0 (0.06 mL, 0.50 mmol). Purification was performed by column chromatography using hexane / EtOAc (95: 5) and 14 (80 mg, 48%) was obtained as an orange solid, mp 187.3-187.8; ^X H MR (700 MHz, CDC1 ₃ ): δ 6.83 (1 H, S, H-7), 2.53 (3 H, s, CH ₃ ), 2.41 (3 H, s, CH ₃ ), 2.36 (2 H , q, J = 7.7 Hz, CH ₂ ), 2.28 (3 H, s, CH ₃ ), 1.00 (3 H, t, J = 7.7 Hz, CH ₃ CH ₂ ); ¹³ C NMR (176 MHz, CDC1 ₃ ): δ 163.7 (C), 141.8 (C), 137.8 (C), 136.6 (2 C), 134.7 (C), 131.7 (C), 130.7 (C), 119.5 ( CH), 116.2 (C), 17.1 (CH ₂ ), 15.8 (CH ₃ ), 14.4 (CH ₃ ), 14.2 (CH ₃ ), 13.3 (CH ₃ ); IR (pure): 1579, 1404, 1206, 1147, 1093, 1022, 801 Cffl ^"1 ; HRMS-EI: calculated for (C ₁₄ H ₁₅ BCl ₂ F ₂ N ₂ ) 330.0668 found 330.0670.

B. EVALUATION OF THE NEW COLORS

The evaluation of the compounds synthesized as laser dyes is described below. Selected pumping conditions are: transverse pump configuration at 532 nm, 10 Hz repetition rate and a pulse energy of 5 mJ. The laser action of each dye is characterized by determining its laser efficiency (Eff}, defined as the relationship between the pumping energy on the sample and the output energy of the dye, the wavelength (λι ₃ ) corresponding to the maximum of its band of emission and its photo stability defined as the% of laser emission that the dye has left after 100,000 pumping pulses at 5 mJ and 10 Hz, always pumping the same region of the sample, so the analysis is done by entering 10 μΐ of dye dissolution in a capillary The laser characteristics of the new dyes pumped in the experimental conditions detailed above are specified in the following tables.

Chlorinated 8-tolyl-BODIPYs

1.- The solutions are in ethyl acetate and the detailed concentrations are those that optimize the laser efficiency (a study of the dependence of its laser efficiency on the concentration has been made for all). 2. - While the non-chlorinated derivative has no laser emission, the presence of chlorine leads to dyes with a highly efficient and stable laser emission.

3. - The efficiency and laser stability of these dyes increases as the number of chlorine atoms in the chromophore increases. 4. - The behavior of these new dyes in different solvents is studied.

Other chlorinated BODIPY

Laser dependence on chlorination and solvent nature.

Compound dye 8 Compound 9

 [c] = 1 mM [c] = 0.8 mM

Ef solvent f (%) Stability A Eff (%) Stability Ala

 (nm) (nm)

 EtOAc 14 615 60 18 612 80

 MeOH 17 612

 Acetone 14 615 18 613

CH, C1 ₂ 10 617 19 621 Compound dye 10

 [c] = 1 mM

 Solvent Eff (%) ia (nm) Stability

EtOAc 30 618 100

 MeOH 21 610

 Acetone 23 614

CH ₂ C1 ₂ 25 621

The unchlorinated derivative (11) was pumped at 500 nm corresponding to its maximum absorption. There is no laser emission pumping at the usual pumping lengths such as 355 nm or 532 nm. However, its chlorinated derivatives can be pumped perfectly at 532 nm, since chlorination displaces the maximum absorption towards red (this is an important advantage).

Unchlorinated pyrometene 546 (15), mono- (16) and dichlorinated (17)

The unchlorinated PM546 commercial dye, even at the concentration of 5 x 10 ^~ M, does not emit laser pumped at 532 nm under the same conditions as its chlorinated derivatives. The PM546 must be pumped at 355 nm while its chlorinated derivatives can be pumped at two wavelengths (355 and 532 nm), which represents an important technological advantage.

 * They are data under pumping at 355 nm; the others are registered under pumping at 532 nm

 Compound dye 17

 [c] = 3.5 mM

 Solvent Eff (%) ia (nm) Stability

EtOAc 51 562 100

 MeOH 44 560

 Acetone 38 570

CH ₂ C1 ₂ 49 563

Claims

1. Use of a compound d

where , at least one of the substituents R ¹ , R ^£ , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ is a chlorine atom attached directly to a carbon of the boradiazaindacene system, and the substituents R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ^s and R ⁷ different from chlorine are independently selected from the group consisting of hydrogen, halogen other than chlorine, cyano, alkyl, aryl, alkenyl, alkynyl, alkoxy and heterocycle, with the proviso that R ⁴ is not an alkyl substituted with hydroxyl or acyl as a laser dye or fluorescent marker. 2. Use of a compound of formula I according to claim 1, wherein the compound of formula I comprises at least two chlorine atoms in the positions R ¹ , R ² , R ³ , R ⁵ , R ^s or R '. 3. Use of a compound of formula I according to claim 2, wherein the compound of formula I comprises at least four chlorine atoms in the positions R ¹ , R ² , R ³ , R ⁵ , R ^s or R ⁷ . 4. Use of a compound of formula I according to any one of claims 1 to 3, wherein said compound acts as a fluorescent marker in an image and / or analysis technique based on laser induced fluorescence emission, and said compound of formula I it is covalently linked to a biomolecule selected from the group consisting of an amino acid, peptide, protein, lipid, carbohydrate, antibody, nucleic acid and toxin. 5. Use of a compound of formula I according to any one of claims 1 to 3 as a laser dye in a laser system used in an analytical, photoelectronic or biophotonic technique. 6. A fluorescent compound of formula I

 I where (a) R ⁴ is an aryl group, R ^" , R ³ , R ⁵ and R ⁷ are independently selected from the group consisting of hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl and heterocycle, at least one of the substituents R ^A or R ^s is a chlorine atom and the other substituent, if it is not chlorine, is independently selected from the group consisting of hydrogen, halogen other than chlorine, alkyl, aryl and heterocycle; or (b) R ⁴ is an unsubstituted alkyl group, R ¹ , R ³ , R ⁵ and R 'are independently selected from the group consisting of halogen, alkyl, alkenyl, alkynyl, aryl and heterocycle, at least one of the substituents R ² or R ⁶ is a chlorine atom and the other substituent, if not chlorine, is independently selected from the group consisting of hydrogen, halogen other than chlorine, alkyl, aryl and heterocycle; or (c) R ⁴ , R °, R ^s and R ⁷ are the same or different alkyl groups, at least one of the substituents R ¹ , R ² or R ³ is a chlorine atom and the other substituents, if not chlorine , be independently select from the group consisting of hydrogen, alkyl, aryl and heterocycle. 7. A fluorescent compound of formula I according to claim 6 option (a), wherein R ⁴ is a toline group. 8. A fluorescent compound of formula I according to claim 7, wherein R ¹ , R ³ , R ⁵ and R 'are independently selected from the group consisting of hydrogen, halogen and monocyclic aryl. 9. A fluorescent compound of formula I according to claim 7 selected from the group comprising 2, 3, 5, 6, 7, 9 and 10.

10. A fluorescent compound of formula I according to any one of claims 6 option (a) or 7, wherein RR ² , R ⁶ and R ⁷ are chlorine atoms. 11. A fluorescent compound of formula I according to claim 6 option (a), wherein R, R ² , R ³ , R °, R ⁶ and R ⁷ are chlorine atoms. 12. A fluorescent compound of formula I according to claim 6 option (b), wherein R ⁴ is a C1-C10 alkyl and R ¹ , R ³ , R ⁵ and R ⁷ are methyl groups. 13. A fluorescent compound of formula I according to claim 12 selected from the group comprising 16 and 17.

 16 17 14. A fluorescent compound of formula I according to claim 6 option (c), wherein R ⁴ , RR ⁶ and R7 ⁴ are the same or different C1-C4 alkyl groups, R ¹ is chlorine, and R ² and R ³ are selected from independent of the group consisting of hydrogen and chlorine. 15. A fluorescent compound of formula I according to claim 14 selected from the group consisting of 12, 13 and 14.

12 13 14 16. A process for obtaining a compound of formula I as described in any one of the claims 6 option (a) or (b), 7 to 13, characterized in that it comprises a chlorination reaction between a compound of formula II and a chlorinating agent in the presence of an organic solvent,

where less a substituent R ^{2 '} or R ^s' is a hydrogen atom directly attached to the boradiazaindacene system and the other substituent, if not hydrogen, is independently selected from the group consisting of halogen, alkyl, aryl and heterocycle, R ^{1 '} , R ^3' , R ^{5 '} and R ^7' are independently selected from the group consisting of hydrogen, halogen, alkenyl, alkynyl, aryl and heterocycle, and R ^{4 '} is an aryl or unsubstituted alkyl group. 17. Method of obtaining a compound of formula I according to claim 16 wherein the chlorinating agent is iV-chlorosuccinimide. 18. Process for obtaining a compound of formula I according to any one of claims 16 or 17 wherein the solvent used is tetrahydrofuran.