Terbium and dysprosium complexes with optimized antenna, useful as luminescent markers

Abstract

The present invention relates to luminescent lanthanide complexes comprising a chelating agent, formed of a macrocycle or a ligand, complexing a lanthanide ion, as well as corresponding chelating agents and a method for detecting a biomolecule using said lanthanide complex.

Description

According to another embodiment, the lanthanide (II) complexes, named (Ile), are 10 such that: -Chrom4- and -Chromé- are identical and have the same structure (B12): with -RI5, -R25, -R35, -R45 and -R5S as defined respectively for -RI, -R2, -R3, -R4 and -R5 in the formula groups (B), it being understood that -Chrom4- and 15 -Chrome- do not include a reactive group, and -Chrom5- is different from -Chrom4- and -Chromé- and has a structure of (B13): (B13) with -RI6, -R26, -R36, -R46 and -R56 as defined respectively for -RI, -R2, -R3, -R4 and -R5 in the formula groups (B), it being understood that -Chrom5- includes a reactive group -XI. Advantageously, in complexes (Ilb) and (Ile), -RI3 and -RI3, when present, are -O(C1-C6) alkyl or -NH(CO)(C1-C6) alkyl groups optionally substituted by a -Y group. Preferably in complexes (Ilb) and (Ile), -RI3 and -RI5 are -O(C1-C6) alkyl groups optionally substituted by a -Y group, and in particular -OMe or -OPEG. Advantageously in complexes (Ilb) and (Ile), -RI4 and -RI6, when present, represent an -O(C1-C6) alkyl or NH(CO)(C1-C6) alkyl group substituted by an -XI group. Preferably in complexes (Ilb) and (Ile), -RI4 and -RI6 are -O(C1-C6) alkyl groups optionally substituted by an -XI group. In complexes (Ilb) and (Ile), R23, -R24, -R25 and -R26, when present, may be identical or different, and represent a group -R7 as defined for groups of formula (B). Preferably in complexes (Ilb) and (Ile), R23, -R24, -R25 and -R26, when present, are identical and represent, preferably -Me. In complexes (Ilb) and (Ile), R23, -R24, -R23, and -R26, when present, may be identical or different and represent a hydrogen or an -R8 group as defined for groups of formula (B), preferably -Me. Preferably, R23, -R24, -R25, -R26, when present, are identical and represent, preferably, a hydrogen. In complexes (Ilb) and (Ile), R33, -R34, -R35 and -R36, when present, may be identical or different and represent a hydrogen or a -R8 group as defined for groups of formula (B), preferably -Me. Preferably, R33, -R34, -R35 and -R36, when present, are identical and represent a hydrogen. Preferably in complexes (Ilb) and (Ile), -R33, -R34, -R33 and -R36, when present, are identical and advantageously represent hydrogens, and -R23, -R24, -R25 and -R26, when present, are identical and each represent an -R7 group, as defined for groups of formula (B), preferably -Me. In particular, Chrom4-, -Chrom5- and -Chromô- are identical or different, and chosen independently of each other from the following formula groups (B14): with -R7 and -El as defined for formula groups (B), and -R7 preferably representing -Me and -El preferably representing a -OR10 or -NCO(NR10) group possibly substituted by a -Y or -XI group, RIO being as defined for formula groups (B). According to a particular embodiment of the invention, the lanthanide complexes (Ilb) are such that: -RI3 is an -O(C1-C6) alkyl group possibly substituted by a -Y group, -RI4 is an -O(C1-C6) alkyl group substituted by an -XI group, -R23 and -R24 are identical or different, preferably identical, and represent an -R7 group, and in particular an (C1-C6) alkyl group, and - -R33, R34, -R43, -R44, -R53 and -R54 are hydrogens. According to a particular embodiment of the invention, the lanthanide complexes (Ile) are such that: -RI5 is an -O(C1-C6) alkyl group possibly substituted by a -Y group, -RI6 is an -O(C1-C6) alkyl group substituted by an -XI group, -R25 and -R26 are identical or different, preferably identical, and represent an -R7 group, and in particular an (C1-C6) alkyl group, and - -R35, R36, -R45, -R46, -R55 and -R56 are hydrogens. The invention also relates to chelating agents corresponding to the lanthanide compounds defined within the scope of the invention, regardless of their described embodiment. The invention also relates to such chelating macrocycles in their protected form according to formula (III) and to such chelating ligands in their protected form according to formula (IV). The invention also relates to chelating agents of formula (III): Chroml ' IN—Chrome2 Chrom3' (ni) with -Chroml', -Chrom2', and -Chrom3', identical or different, and chosen independently from each other from the formula groups (B5): with RI, R2, R3, R4 and R5 as defined for lanthanide (I) complexes, and -R13 a protecting group of acid functions (i.e. -Z(O)OH), such as an alkyl, and in particular a methyl, or in the form of a salt. The invention also relates to chelating agents of formula (IV): R11. RaGOC^N , Chrom4 ri 2 N^'COORa Chrom6 Chrom5 R13COO COOR13 (IV) with -Chrom4-, -Chrom5- and -Chrome- as defined for lanthanide complexes of formula (II), and -R13 a protecting group of acid functions, such as an alkyl, and especially a methyl, or in the form of a salt. The preparation of complexes (I) and (II) according to the invention employs conventional techniques and reactions known to those skilled in the art. In particular, they can be obtained using methods similar to those described in the examples. Complexation reactions are generally carried out with a salt of the desired lanthanide, in particular a chloride, a nitrate or a triflate, in a dissociating solvent, in particular an alcohol such as methanol, THF or acetonitrile, or in a mixture of solvents, usually in the presence of a carbonate, at a temperature between 25 and 80 °C, for a duration ranging from 30 minutes to a few hours. Compounds (III) and (IV) are relatively easy and conventional to obtain, and can be produced at a reasonable cost. The reagents are commercially available or readily accessible. The functional groups that may be present in compounds of formulas (III) and (IV), and in reaction intermediates, can be protected during the syntheses, either permanently or temporarily, by protecting groups that ensure the unambiguous synthesis of the expected compounds. The protection and deprotection reactions are carried out according to techniques well known to those skilled in the art. (By protecting group) temporary, we mean protecting groups such as those described in Protective Groups in Organic Synthesis, Greene TW and Wuts PGM, ed. John Wiley & Sons, 2006, and in Protecting Groups, Kocienski PJ 1994, Georg Thieme Verlag. Formula (I) describes the family of lanthanide complexes resulting from the coordination of a macrocyclic ligand of formula (III) after deprotection and release of the acid functions -Z(O)OH (dialkyl acid or phosphinic diaryl when -Z- = -P(Rz)- or carboxylic acid when -Z- = -C-). According to a first embodiment, -Chrom1', -Chrom2', and -Chrom3' are identical. According to this embodiment, the chelating agent of formula (III) can be obtained by alkylation of a TACN.3HC1 type ring with an intermediate of formula (B6): with -A-, -RI, -R2, -R3, -R4, -R5 and -R13 as defined for chelating agents (III), and -LG representing a departing group. A "leaving group" is a group capable of detaching from the rest of the molecule under the reaction conditions, so that another reactant can react with the remaining molecule to form a new bond with the carbon atom initially bearing the leaving group. The nature of the leaving group is not limited, and any leaving group known to those skilled in the art can be used. Examples include sulfonates, such as mesylate or tosylate. The intermediate (B6) can be prepared, using techniques well known to those skilled in the art, from the following intermediate (B7): R1 OR13 0R13 (B7) with -RI, -R2, -R3, -R4, -R5, -R13 and -Z- as defined for the intermediate (B6). The intermediate (B7) can be synthesized from the reagent (B9), by formation of a boronic acid (B8), followed by a pyridinedicarboxylate reaction: Suzuki with a dialkyl-4-halogeno-2,6- R1 £ —* (B7) B(OR)2 : defined for the intermediate (B7) and -hated with -RI, -R2, -R3, -R4 and -R5 such as represent a halogen, like -Br, -Cl or -I. According to a second embodiment, -Chroml' and -Chrom2' are identical, and -Chrom3' is different from -Chroml' and -Chrom2'. According to this embodiment, the chelating agent of formula (III) can be prepared by alkylation of the intermediate (III') with the intermediate of formula (B6”), with -RI”, -R2”, -R3”, -R4”, -R5”, -R13”, -Z”, and -LG”, as defined respectively for -RI, -R2, -R3, -R4, -R5, -R13, -Z, and -LG for the chelating agent of formula (III): RI" LG" OR|3" The intermediate (III') can be prepared by alkylation of a TACN.3HC1 type ring with the intermediate of formula (B6'), with -RI', -R2', -R3', -R4', -R5', -R13', -Z' and -LG' as defined respectively for -RI, -R2, -R3, -R4, -R5, -R13, -Z and -LG for the chelating agent (III): R1 Advantageously, one of the three amine functions of the TACN.3HC1 type ring is protected, via an amine protecting group, prior to the alkylation step. with the intermediate of formula (B6'); a subsequent deprotection step allowing the generation of the intermediate (III'). The intermediates (B6') and (B6”) can be obtained in a manner analogous to the intermediate (B6). Formula (II) describes the family of lanthanide complexes resulting from the coordination of a chelating ligand of formula (IV) after deprotection and release of the carboxylic acid groups. For the synthesis of these compounds, reference may be made in particular to Takalo applications WO 2013 / 026790 and WO 2013 / 092992. The invention finally relates to a method for detecting a biomolecule comprising the detection of the luminescence of a conjugate of said biomolecule with a luminescent complex as defined within the framework of the invention, comprising a reactive group, and obtained by coupling said biomolecule with said luminescent complex on its reactive group. Figure 1 shows the absorption and emission spectrum of the Ia.4 complex in water at room temperature. Figure 2 shows the emission spectrum of complex Ia.6 in a 4:1 methanol / ethanol mixture at room temperature. Figure 3 shows images of cells fixed to PFA and labeled with complex Ia.2 (two-photon excitation Xex = 720 nm), obtained with visible emission of complex Ia.2 (left image) or obtained in transmitted DIC light (right image). Figure 4 shows images of cells fixed to PFA and labeled with complex Ia.5 (two-photon excitation Xex = 720 nm), obtained with visible emission of complex Ia.5 (images a and c) or obtained in transmitted DIC light (images b and d). The examples below are intended to illustrate the invention, but are not exhaustive. Examples The following abbreviations are used in the examples that follow: DCM: dichloromethane; DMF: dimethylformamide; DMSO: dimethyl sulfoxide; Ms: mesyl; nd: non-detenninized; PBS: phosphate-buffered saline; PEG: polyethylene glycol; PFA: paraformaldehyde; quant.: quantitative; TACN: triazacyclononane; THF: tetrahydrofrequency; Ts: tosyl General Information: The ¹³C (¹H, ¹³C) NMR spectra were recorded on a Broker Avance instrument at frequencies of 300 MHz or 500.10 MHz, and 75 MHz or 125.75 MHz, for ¹H and ¹³C respectively. Chemical shifts (σ) are expressed in parts per million (ppm) relative to trimethylsilane used as an internal reference and using the solvents indicated. Coupling constants (J) are expressed in Hz, and the following notations are used: s (singlet), br (broad), d (doublet), t (triplet), dd (doublet of a doublet), m (multiplet). High-resolution mass spectra were recorded at the Lyon Joint Mass Spectrometry Center (Université Claude Bernard Lyon, Lyon, France) on a MicroTOFQII instrument equipped with a positive ESI source. Thin-layer chromatography was performed on silica gel plates on aluminum foil (Fluka) and visualized using a UV lamp (X = 254 or 365 nm) or by staining. Purifications were carried out using a silica gel chromatography column (0.035–0.070 mm, 60 Å). The solvents used for the reactions were purchased from Aldrich or Acros Organics as dry or extra dry solvents and stored on 3Å molecular sieve and the reagents used were purchased from Aldrich, Acros Organics or Alfa Aesar.UV / Vis absorption spectra were recorded on a JASCO V670 spectrometer; emission spectra on a JOBIN-YVON fluorolog 3 spectrofluorimeter. Retention times were performed on an Agilent Technologies 1260 instrument equipped with a Waters XBridge RP-C18 column (3.5 µm, 4.6 x 100 mm). The chromatographic system used was 0.25M ammonium formate-MeCN (v / v) as eluents: isocratic 15% MeCN (2 min), linear gradient 15 to 100% MeCN (16 min), isocratic 100% MeCN (4 min), at a flow rate of 1 mL min⁻¹, with UV detection at 210 and 252 nm. Cell imaging assays were performed using a concentrated solution of the complex in DMSO to obtain a final complex concentration of approximately 1 x 10³ M in the cell culture medium (PBS) with <1% DMSO. T24 cells were pre-fixed to PFA. Two-photon absorption spectra were acquired using an LSM710NLO laser scanning confocal microscope (CarlZeiss). A. PREPARATION DES COMPLEXES Exemple 1 : Complexes d’Europium la-Ie Schéma 1 : Synthèse des intermédiaires 5a-5e PdCI2(dppf)2, AcOK DMSO B(OR)2 2a R=H, R^OMe, R2=Me, R3=H 2b R=C(CH3)2, R^OPeg, R2=Me, R3=H (90%) 2c R=C(CH3)2. R^NHCOMe, R2=Me, R3=H (50%) 2d R=C(CH3)2, R^OPeg, R2=R3=Me (68%) 2e R=H, R1=R2=OMe, R3=H Pd(PPh3)4 / CsF (a,d) Pd(dba)3tBu3PH.BF4 / KF (b,C,e) DMF 5a (85%) 4a (83%) 5b (91%) 4b (90%) 5c (quant.) 4c (58%) 5d (quant.) 4d (98%) 5e (quant.) 4e (56%) 3d (39%) 3e (63%) The compounds 1a (4-bromo-3-methylanisole), 1a (l-bromo-3,5-dimethoxybenzene), and 2e (2,4-dimethoxyphenylboronic acid) are commercially available. Compounds 1b and 1d are prepared from the corresponding phenol, while compound 1e is prepared from the corresponding aniline. Boronic acids 2a-2d are then obtained by a procedure well known to those skilled in the art. Compounds 3a-e are obtained by a Suzuki coupling reaction with dimethyl-4-iododipicolinate. A reduction and mesylation reaction then isolates compounds 5a-e. Diagram 2: Synthesis of the la-Ie complexes NaOH IM,THF Ri=OMe, R2=Me, Rj=H (quant.) ya lb Ri=OPeg, R2=Me, Rq=H (quant) 7^ IC R^NHCOME, R3=Me,R3=H (83%) 7C ld RpOPeg, R2=R3=Me (90%) 7J R1=R2=OMe,R3=H (85%) 7th An alkylation reaction of TACN.3HC1 by mesylate derivatives 5a-e, in classic alkylation conditions, followed by a acid function deprotection step carboxylic, allow the lanthanide complexes la-Ie to be obtained. 4-Bromo-3-methylphenol (1.0 g, 5.3 mmol) and K₂CO₃ (3.7 g, 27 mmol) are dried under vacuum and then solubilized in dry DMF (20 mL). After 30 minutes, triethylene glycol methyl ether tosylate (2.55 g, 8 mmol) is added to the Schlenck. The reaction mixture is stirred at 80°C for 5 days. The solution is then filtered through a P3 sintered filter, rinsed with CH₂Cl₂ MO₂, and concentrated under vacuum. The mixture is diluted in AcOEt / EtyO₂ and washed with H₂O and NaCl. The organic phase is dried on NaySCl, filtered, and evaporated. The crude product is then purified on Colon chromatography (SiO2, CH2Cl2 / AcOEt, 90 / 10 to 80 / 20) to donner the product in sous vide color (1.62 g, 91%). Rf (SiO2, CH2Cl2 / AcOEt, 80 / 20) = 0.53 ; *H NMR (300 MHz, CDC13, ô): 7.38 (d, 1H, J = 8.7 Hz, H6), 6.81 (d, 1H, J = 2.9 Hz, H3), 6.62 (dd, 1H, J3= 8.7 Hz, J4= 2.9 Hz, H5), 4.08 (t, 2H, J= 4.6 Hz, H8), 3.83 (t, 2H, J= 4.6 Hz, H9), 3.75-3.53 (m, 8H, H1c7Hu / H12 / H13), 3.38 (s, 3H, H14), 2.35 (s, 3H, H7) ; 13C NMR (100 MHz, CDC13, ô): 158.2 (C4), 138.9 (C2), 132.9 (C6), 117.4 (C3), 115.8 (C1), 113.8 (C5), 72.1 (C13), 71.0 (C10), 70.8 (C), 70.8 (C12), 69.8 (C9), 67.8 (C8), 59.2 (C14), 23.3 (C7) In a round-bottom flask, acetic anhydride (305 pL, 3.22 mmol) is added to a solution of 4-bromo-3-methylaniline (500 mg, 2.69 mmol) in dry CH2Cl2 under Argon. The mixture is stirred at room temperature under Argon until the reaction is complete (monitored by TLC, petroleum ether / ethyl acetate, 80 / 20). After 4 h, the mixture is washed with H2O (3x). The organic phase is dried over Na2SO4, filtered through cotton, and evaporated to obtain a white solid (506 mg, 82%). Rf (SiO2, CH2Cl2 / AcOEt, 80 / 20) = 0.46; *H NMR (300 MHz, CDC13, ô): 7.45-7.42 (m, 2H, H6 / H3), 7.19 (dd, 1H, J3= 8.6 Hz, J4= 2.4 Hz, H5), 2.36 (s, 3H, H7), 2.16 (s, 3H, H9); 13C NMR (75 MHz, CDC13, ô): 169.0 (C8), 138.5 (C4), 137.2 (C1), 132.6 (C3), 122.4 (C6), 119.6 (C2), 119.2 (C5), 24.5 (C9), 23.1 (C7); HRMS (ESI) calculated for C9Hi,BrNO228.0024. Exp 228.0019 [M+H]+. Compound Id Br Id 4-Bromo-3,5-Dimethylphenol (1.30 g, 6.46 mmol) and K₂CO₃ (4.46 g, 9.68 mmol) are vacuum-dried and then solubilized in dry DMF (12 mL). Argon bubbling is performed for 10 minutes, and triethylene glycol methyl ether tosylate (3.08 g, 9.68 mmol) is added to the Schlenck. The reaction mixture is stirred at 80°C for 3 days. A CH₂Cl₂ / H₂O extraction is then performed, and the organic phase is dried with NaCl and Na₂SO₄, filtered, and evaporated. The crude product is then purified by column chromatography (SiO₂, CH₂Cl₂ / AcOEt, 100 / 0 to 80 / 20) to give 1.5 g of pure product oil (67%). (SiO2, CH2Cl2 / AcOEt, 95 / 5) = 0.38 ; *H NMR (300 MHz, CDC13, ô): 6.66 (s, 2H, H3), 4.07 (t, 2H, J= 4.7 Hz, H8), 3.82 (t, 2H, J= 4.7 Hz, H9), 3.74-3.52 (m, 8H, H10 / Hu / H12 / H13), 3.37 (s, 3H, H14), 2.36 (s, 6H, H7) ; 13C NMR (75 MHz, CDCI3, ô): 157.4 (C4), 139.2 (C1), 118.5 (C2 / 6), 114.7 (C3 / 5), 72.1 (C13), 71.0 (C10), 70.8 (C11), 70.7 (C12), 69.8 (C9), 67.7 (C8), 59.2 (C14), 24.1 (C7) ; HRMS (ESI) calculated for C15H23BrNaO4 369.0672. Exp 369.0664 [M+Na]\ Compose 2a The composition 2a was prepared according to the protocol described in “Substituent Effects on Oxidation-Induced Formation of Quinone Methides from Arylboronic Ether Precursors”, Sheng Cao, Robin Christiansen, and Xiaohua Peng, Chem. Eur. J., 2013, 19, 9050 - 9058. A o. .0. j\ 9 10 ° 13 14 5Q3 6 h crB'o --4-16 Composed 2b / 15\ 2b Dry potassium acetate (442 mg, 8.46 mmol) is dissolved in dry DMSO (15 mL) under argon, then bis(dipicolinato)diboron (572 mg, 2.25 mmol) and 1b (500 mg, 1.50 mmol) are added. The solution is placed under argon bubbling for 20 minutes, then PdCl2(dppf)2 (77 mg, 0.11 mmol) is added. The reaction mixture is heated at 90°C under argon with stirring for 16 h. A serial H2O / Et2O extraction is then performed. The organic phases are collected, washed with NaClSat, dried over Na2SO4, filtered, and evaporated under vacuum. The crude product is then purified on a column (SiO2, CH2Cl2 / AcOEt, 90 / 10 to 80 / 20) to obtain the pure product in the form of yellow oil (516 mg, 90%). Rf (SiO2, CH2Cl2 / AcOEt, 80 / 20) = 0.57; 'H NMR (500 MHz, CDCI3, ô): 7.69 (d, 1H, J = 8.1 Hz, H6), 6.72-6.69 (m, 2H, H3 / H5), 4.13 (t, 2H, J= 4.7 Hz, H8), 3.85 (t, 2H, J= 4.7 Hz, H9), 3.75-3.53 (m, 8H, H10 / Hll / H12 / H13), 3.38 (s, 3H, H14), 2.50 (s, 3H, H7), 1.32 (s, 12H, H16) ; 13C NMR (125 MHz, CDCI3, ô): 161.1 (C4), 147.3 (C2), 138.0 (C6), 120.6 (C1), 116.4 (C3), 110.9 (C5), 83.3 (C15), 72.1 (72.C). (C11), 70.8 (C12), 69.9 (C9), 67.2 (C8), 59.2 (C14), 25.1 (C16), 22.6 (C7) ; HRMS (ESI) calculated for C20H33BNaO6 403.2262. Exp 403.2255 [M+Na]+. . O —4—4—11 Composed of 2c / 10\ 2c Potassium acetate (515 mg, 5.25 mmol) is dried under vacuum / argon for 1 hour, then bis(pinacolato)diboron (668 mg, 2.63 mmol), dry DMSO (6 mL), and [unclear] (400 mg, 1.75 mmol) are added. Argon bubbling is performed in the mixture for 20 minutes, then PdC12(dppf)2.CH2C12 (100 mg, 0.12 mmol) is added and the Schlenck is closed. The solution is heated to 90°C under argon with stirring for 19 hours. After returning to room temperature, the mixture is filtered through P3 sintered glass and rinsed with Et2O. A serial extraction with H2O / Et2O is then performed. The organic phases are collected, dried over Na2SO4, filtered through cotton, and evaporated. The slow evaporation of CH2Cl2 leads to the formation of crystals which are washed several times with CH2Cl2 and dried under vacuum. ¹H NMR shows the obtaining of the expected pure product (240 mg, 50%). Rf (SiO2, CH2Cl2 / AcOEt, 80 / 20) = 0.61; *H NMR (300 MHz, CDC13, δ): 7.72 (d, 1H J= 8 Hz, H5), 7.33 (s, 1H, H3), 7.30 (d, 1H, J= 8 Hz, H6), 7.18 (s, 1H, NH), 2.51 (s, 3H, H7), 2.16 (s, 3H, H9), 1.33 (s, 12H, H11) ; 13C NMR (75 MHz, CDCI3, ô): 168.7 (C8), 146.4 (C2), 140.3 (C4), 137.1 (C6), 120.6 (C3), 115.9 (C5), 83.4 (C10), 25.1 (C9), 25.0 (C11), 22.4 (C7) ; HRMS (ESI) calculée pour C15H23BNO3 276.1768. Exp 276.1762 [M+H]+. . Dry potassium acetate (830 mg, 8.46 mmol) is dissolved in dry DMSO (10 mL) under argon, then bis(dipicolinato)diboron (1.07 g, 4.23 mmol) and Id (979 mg, 2.82 mmol) are added. The solution is placed under argon bubbling for 20 minutes, then PdC12(dppf)2.CH2C12 (161 mg, 0.20 mmol) is added. The reaction mixture is heated at 90°C under argon with stirring for 40 h. A serial H2O / Et2O extraction is then performed. The organic phases are collected, washed with NaClSat, dried over Na2SO4, filtered, and evaporated under vacuum. The crude product is then purified on a chromatographic column (SiO₂, CH₂Cl₂ / AcOEt, 95 / 5 to 80 / 20) to obtain the pure product in the form of oil (748 mg, 68%). ^(SiCh, CH₂Cl₂ / AcOEt, 80 / 20) = 0.43; *H NMR (300 MHz, CDC13, θ): 6.51 (s, 2H, H₃), 4.09 (t, 2H, J= 4.7 Hz, H₈), 3.82 (t, 2H, J= 4.7 Hz, H₂9), 3.74-3.53 (m, 8H, H₁₀ / H₁₂ / H₁₃), 3.37 (s, 3H, H₁₄), 2.37 (s, 6H, H₂7), 1.36 (s, 12H, H16) ; 13C NMR (75 MHz, CDCI3, ô): 159.7 (C4), 144.5 (C2'6), 113.2 (C3 / 5), 83.5 (C15), 72.1 (C13), 70.9 (C10), 70.8 (C11), 70.7 (C12), 69.9 (C9), 67.1 (C8), 59.1 (C14), 25.0 (C16), 22.7 (C7) ; HRMS (ESI) calculated for C2iH35BNaO6417.2423. Exp 417.2417 [M+Na]+. . Compose 3a 3rd In a nutshell, diméthyl-4-iododipicolinate (300 mg, 0.934 mmol) and acid 2-Methyl,4-Methoxyphenylboronic acid (186 mg, 1.121 mmol) is dissolved in dry DMF (15 mL) under argon. After 30 minutes of argon bubbling, cesium fluoride (355 mg, 2.335 mmol) and Pd(PPh3)4 (cat.) are added. The reaction mixture is stirred at 90°C under argon for 17 h. The mixture is then diluted in CH2Cl2 and extracted with H2O. The chlorinated phase is evaporated, and the aqueous phase is extracted again with AcOEt. The organic phase is washed with NaCl, dried over Na3SO4, filtered, and evaporated under vacuum. The crude product obtained is purified on 2 chromatographic columns (SiCl, CH2Cl2 / acetone, 90 / 10 then SiCh, CH2Cl2 / AcOEt 98 / 2 to 95 / 5) to obtain the pure product (159 mg, 54%). Æ / (SiO2, CH2Cl2 / acetone, 80 / 20) = 0.82; *H NMR (300 MHz, CDC13, δ): 8.28 (s, 2H, H35), 7.24-7.20 (m, 1H, H12), 6.87-6.85 (m, 2H, Hn / H9), 4.04 (s, 6H, H16), 3.86 (s, 3H, H14), 2.32 (s, 3H, H13); 13C NMR (125 MHz, CDCI3, ô): 165.5 (C15), 160.5 (C10), 152.6 (C4), 148.4 (C“6), 137.0 (C8), 131.0 (C12), 128.8 (C7), 121.7 (C3 / 5), 116.6 (C9), 112.1 (C11), 55.5 (C14), 53.4 (C16), 20.8 (C13); HRMS (ESI) calculated C17H17NNaO5 338.0999 Exp 338.0990 [M+Na]+. Compound 3b OO 3b Dry dimethyl 4-iodo-2,6-pyridinedicarboxylate (344 mg, 1.07 mmol) is dissolved in dry DMF (12 mL), and 2b (448 mg, 1.18 mmol) is added under argon. The solution is placed under argon bubbling for 30 minutes, then potassium fluoride (205 mg, 3.53 mmol) and Pd(dba)3tBu3PH.BF4 (43 mg, 0.08 mmol) are added. The reaction mixture is placed at 80°C under stirring and argon for 40 h. A serial AcOEt / H2O extraction is then performed. The organic phases are washed with NaClsat, dried over Na2SC>4, filtered, and evaporated under vacuum. The crude product is then purified on a chromatographic column (SiO₂, CH2C12 / acetone, 95 / 5 to 90 / 10) to obtain the pure product in the form of a white solid (343 mg, 72%). R / (SiO2, CH2Cl2 / acétone, 90 / 10) = 0.45 ; XH NMR (500 MHz, CDC13, ô): 8.26 (s, 2H, H35), 7.19 (d, 1H, J = 8.3 Hz, H12), 6.87 (d, 1H, J = 2.3 Hz, H9), 6.86 (dd, 1H, J3= 8.3 Hz, J4= 2.3 Hz, H11), 4.17 (t, 2H, J = 4.7 Hz, H14), 4.03 (s, 6H, H22), 3.86 (t, 2H, J = 4.7 Hz, H15), 3.76-3.54 (m, 8H, H16 / Hl7 / H!8 / H19), 3.38 (s, 3H, H20), 2.30 (s, 3H, H13) ; I3C NMR (125 MHz, CDC13, ô): 165.5 (C21), 159.7 (C10), 152.6 (C4), 148.3 (C26), 136.9 (C8), 130.9 (C12), 130.1 (C7), 128.7 (C3 / 5), 117.3 (C9), 112.7 (C11), 72.1 (C19), 71.0 (C16), 70.9 (C17), 70.8 (C18), 69.8 (C15), 67.7 (C14), 59.2 (C20), 53.4 (C22), 20.8 (C13) ; HRMS (ESI) calculée pour C23H3ÜNO8 448.1966. Exp 448.1959 [M+H]+. HN 14 wl Composé 3c 2C (200 mg, 0.73 mmol) and dimethyl 4-iodo-2,6-pyridinedicarboxylate (212 mg, 0.66 mmol) are pre-dried under vacuum / argon, then 15 mL of dry DMF is added. The solution is degassed for 20 min under argon, and potassium fluoride (126 mg, 2.18 mmol) and Pd(dba)3tBu3PH.BF4 (27 mg, 0.05 mmol) are added. The mixture is placed at 80°C under argon for 3 days. After returning to room temperature, CH2Cl2 is added, and the organic phase is washed with H2O (3 times) and saturated NaCl (1 time). The organic phase is dried over Na2SO4, filtered, and evaporated under vacuum. After slow evaporation of a DCM / MeOH mixture, crystals are obtained and rinsed with a minimum of CH2Cl2 to obtain the pure product (107 mg, 47%). Rf (SiO2, CH2Cl2 / MeOH, 95 / 5) = 0.55 ; ’fl NMR (300 MHz, CDCI3, ô): 8.25 (s, 2H, H3'5), 7.96 (s, 1H, NH), 7.55 (s, 1H, H9), 7.48 (d, 1H, J= 8.3 Hz, H11), 7.19 (d, 1H, J- 8.3 Hz, H12), 4.00 (s, 6H, H17), 2.26 (s, 3H, H13), 2.19 (s, 3H, H15) ; 13C NMR (75 MHz, CDC13, ô): 168.9 (C14), 165.3 (C16), 152.3 (C4), 148.3 (C2Z6), 139.2 (C10), 136.2 (C8), 133.0 (C7), 130.2 (C12), 128.6 (C3 5), 121.9 (C9), 117.8 (C11), 53.3 (C17), 24.7 (C15), 20.6 (C13) ; HRMS (ESI) calculée pour C18H19N2O5 343.1288. Exp 343.1289 [M+H]+. Composé 3d 20 3d Dry dimethyl 4-iodo-2,6-pyridinedicarboxylate (144 mg, 0.45 mmol) is dissolved in dry DMF (5 mL), and 2d (195 mg, 0.50 mmol) is added under argon. The solution is placed under argon bubbling for 20 minutes, then cesium fluoride (171 mg, 0.11 mmol) and Pd(PPh3)4 (52 mg, 0.05 mmol) are added. The reaction mixture is placed at 80°C under stirring and argon for 5 days. A serial AcOEt / H2O extraction is then performed. The organic phases are washed with NaClSat, dried over Na2SO4, filtered, and evaporated under vacuum. The crude product is then purified on a chromatographic column (SiO2, CH2Cl2 / acetone, 100% to 80 / 20) to obtain the pure product in the form of oil (85 mg, 39%). Æ / (SiO2, CH2Cl2 / acetone, 95 / 5) = 0.41; 'H NMR (300 MHz, CDC13, ô): 8.08 (s, 2H, H3 / 5), 6.66 (s, 2H, H8), 4.11 (t, 2H, J- 4.6 Hz, H14), 3.99 (s, 6H, H22), 3.83 (t, 2H, J= 4.6 Hz, H15), 3.72-3.50 (m, 8H, H16 / H17 / H18 / H19), 3.34 (s, 3H, H20), 1.94 (s, 6H, H13) ; 13C NMR (75 MHz, CDCl3, δ): 165.2 (C21), 158.7 (C10), 152.5 (C4), 148.7 (C2\ 136.5 (C8 / 12), 130.0 (C7), 129.6 (C3 / 5), 114.0 (C9 / 11), 72.0 (C19), 70.9 (C16), 70.7 (C17), 70.6 (C18), 69.7 (C15), 67.4 (C14), 59.0 (C20), 53.2 (C22), 21.0 (C13). . Compound 3e In a Schlenck flask, 4-dimethyliododipicolinate (200 mg, 0.623 mmol) and 2,4-dimethoxyphenylboronic acid (125 mg, 0.685 mmol) are dissolved in 7 mL of dry DMF under argon. After 30 minutes of argon bubbling, potassium fluoride (119 mg, 2.056 mmol) and Pd(dba)3tBu3PH.BF4 (36 mg, 0.062 mmol) are added. The reaction mixture is stirred at 80°C under argon for 19 h. The mixture is then diluted in an AcOEt / Et2O mixture and extracted with H2O and then NaCl. The organic phase is then dried over Na2SO4, filtered, and evaporated under vacuum. The resulting brown solid is then purified using a chromatographic column (SiO2, AcOEt / EP, 20 / 80 to 60 / 40, solid deposit) to obtain the produced in beige solid forms (131 mg, 63%). (SiO2, EP / AcOEt, 60 / 40) = 0.73 ; 'H NMR (300 MHz, CDC13, ô): 8.50 (s, 2H, H3 / 5), 7.40 (d, 1H, J = 8.5 Hz, H12), 6.63 (dd, 1H, J3= 8.4 Hz, J4= 2.4 Hz, H11), 6.58 (d, 1H, J = 2.4 Hz, H9), 4.04 (s, 6H, H16), 3.88 (s, 3H, H13), 3.87 (s, 3H, H14) ; 13C NMR (100 MHz, CDCI3, Ô): 165.8 (C15), 162.5 (C10), 158.2 (C8), 149.2(C4), 148.1(C2'6), 131.6(C12), 128.4(C3 / 5), 118.6(C7), 105.5(C9), 99.2(Cn), 55.8(C13), 55.7(C14), 53.4 (C16) ; HRMS (ESI) calculated for Ci7H17NNaO6 354.0948. Exp 354.0937 [M+Na]+. Compose 4a 0 4th 3a (180 mg, 0.571 mmol) is dissolved in a CH2Cl2 / MeOH mixture (2 / 3 mL) for refroids 0°C. NaBH4 (42 mg, 1.1 mmol) is then added and after 30 minutes, the reaction mixture is placed at room temperature for 4 h. The reaction is then quenched with an IM HCl solution. The organic phase is washed with H2O (2x) then NaClsat, dried over Na2SO4, filtered and evaporated. The crude product is then purified on a chromatographic column (SiO2, CH2Cl2 / acetone 80 / 20 to 65 / 35) to obtain the pure product in the form of pale yellow crystals (136 mg, 83%). Rf (SiO2, CH2Cl2 / acetone, 80 / 20) = 0.25; *H NMR (500 MHz, CDC13, ô): 8.01 (s, 1H, H3), 7.46 (s, 1H, H5), 7.17 (d, 1H, J = 8.8 Hz, H12), 6.85-6-81 (m, 2H, Hn / H9), 4.89 (d, 2H, J = 4.6 Hz, H17), 4.01 (s, 3H, H16), 3.85 (s, 3H, H14), 3.31 (t, 1H, J = 4.6 Hz, OH), 2.29 (s, 3H, H13); 13C NMR (125 MHz, CDC13, ô): 165.9 (C15), 160.1 (C6, C10), 151.8 (C4), 147.1 (C2), 136.9 (C8), 130.9 (C7), 130.8 (C12), 125.0 (C3), 124.6 (C5), 116.5 (C9), 111.9 (C11), 64.9 (C17), 55.5 (C14), 53.1 (C16), 20.8 (C13); HRMS (ESI) calculated for Ci6H]8NO4 288.1230. Exp 288.1220 [M+H]+. Wedge for Ci6Hi7NNaO4 310.1050. Exp310.1038 [M+Na]+. . Compound 4b 4b 3b (320 mg, 0.72 mmol) is solubilized in a CH2Cl2 / MeOH mixture (2.5 mL / 4.5 mL). After placing the medium at 0°C, NaBH4 (30 mg, 0.79 mmol) is added and after 30 minutes, the The solution is brought to room temperature with stirring for 3 hours. The reaction is quenched by adding an IM HCl solution. Then, a CH2Cl2 / H2O extraction is performed. The organic phase is washed with NaCl Sat, dried over Na2SO4, filtered, and evaporated. Purification is performed on a column. Chromatographic analysis (SiO2, CH2Cl2 / MeOH, 98 / 2 to 97 / 3) is carried out to obtain the pure product in the form of a colorless oil (270 mg, 90%). Rf (SiO2, CH2Cl2 / MeOH, 97 / 3) = 0.22; *H NMR (500 MHz, CDC13, ô): 7.99 (d, 1H, J = 0.9 Hz, H3), 7.47 (s, 1H, H5), 7.15 (d, 1H, J = 8.4 Hz, H12), 6.85 (d, 1H, J = 2.3 Hz, H9), 6.83 (dd, 1H, J3= 8.4 Hz, J4= 2.3 Hz, H11), 4.89 (d, 2H, J = 3.9 Hz, H23), 4.16 (t, 2H, J= 4.7 Hz, H14), 3.99 (s, 3H, H22), 3.87 (t, 2H, J- 4.7 Hz, H15), 3.72-3.54 (m, 8H, H16 / H17 / H18 / H19), 3.51 (m, 1H, OH), 3.37 (s, 3H, H20), 2.27 (s, 3H, H13); I3C NMR (125 MHz, CDC13, ô): 165.9 (C21), 160.3 (C6), 159.3 (C10), 151.8 (C4), 147.1 (C2), 136.8 (C8), 131.0 (C7), 130.8 (C12), 125.0 (C3), 124.6 (C5), 117.2 (C9), 112.5 (C11), 72.1 (C19), 71.0 (C16), 70.9 (C17), 70.8 (C18), 69.9 (C15), 67.6 (C14), 64.9 (C23), 59.2 (C20), 53.1 (C22), 20.8 (C13); HRMS (ESI) calculated for C22H30NO7 420.2017. Exp 420.2015 [M+H]+. Compound 4c 0 4c 3c is solubilized in a CH2C12 / MeOH mixture and NaBH4 is added at 0°C. After After 5 minutes, the solution is left at room temperature and the monoreduction is monitored by TLC (SiO2, DCM / MeOH, 95 / 5). After 2.5 hours, the reaction is quenched with IM HCl. The organic phase is then washed with H2O and NaCl, then dried over Na2SO4, filtered, and evaporated. The crude product is purified on a chromatographic column (SiO2, DCM / MeOH, 95 / 5) and a white powder is obtained (56 mg, 58%). Φ / (SiO2, CH2Cl2 / MeOH, 95 / 5) = 0.30; NMR (300 MHz, CDCI3, ô): 7.98 (s, 1H, H3), 7.48 (m, 2H, H4 / H9 / NH), 7.42 (d, 1H, J= 8.3 Hz, H11), 7.17 (d, 1H, J= 8.3 Hz, H12), 4.90 (s, 2H, H18), 4.00 (s, 6H, H17), 3.57 (s, 1H, OH), 2.26 (s, 3H, H13), 2.20 (s, 3H, H15); 13C NMR (75 MHz, CDC13, ô): 168.7 (C14), 165.8 (C16), 160.4 (C6), 151.4 (C4), 147.1 (C2), 138.6 (C10), 136.2 (C8), 134.2 (C7), 130.2 (C12), 124.7 (C3), 124.4 (C5), 121.9 (C9), 117.7 (C11), 64.9 (C18), 53.1 (C17), 24.8 (C15), 20.6 (C13); HRMS (ESI) calculated for C17Hi9N2O4315.1339. Exp 315.1342 [M+H]+. Compound 4d O 4d 3d (136 mg, 0.29 mmol) is solubilized in a CH2Cl2 / MeOH mixture (50 / 50, 4mL). After placing the medium at 0°C, NaBH4 (42 mg, 1.11 mmol) is added (the solution turns brown) and after 20 minutes, the solution is brought to room temperature with stirring for 6 hours. The reaction is quenched by adding an IM HCl solution (4 mL). Then, a CH2Cl2 / H2O extraction is performed. The organic phase is washed with NaCl Sat, dried over Na2SO4, filtered, and evaporated. Purification is carried out on a chromatographic column (SiO2, CH2Cl2 / AcOEt, 70 / 30 to 50 / 50 then 10% MeOH) to obtain the pure product (123 mg, 98%). Δ / (SiO2, CH2Cl2 / AcOEt, 80 / 20) = 0.32; *H NMR (300 MHz, CDC13, ô): 8.08 (s, 1H, H3), 7.79 (s, 1H, H5), 6.63 (s, 2H, H9), 4.87 (s, 2H, H23), 4.09 (t, 2H, J= 4.6 Hz, H14), 3.93 (s, 3H, H22), 3.81 (t, 2H, J= 4.6 Hz, H15), 3.72-3.49 (m, 8H, H16 / H17 / H18 / H19), 3.33 (s, 3H, H20), 1.93 (s, 6H, H13); 13C NMR (75 MHz, CDC13, Ô): 165.6 (C21), 161.0 (C6), 158.2 (C10), 151.4 (C4), 147.1 (C2), 136.4 (C8 / 12), 130.9 (C7), 125.2 (C3 / 5), 113.6 (C9 / n), 71.8 (C19), 70.7 (C16), 70.5 (C17), 70.4 (C18), 69.6 (C15), 67.2 (C14), 64.6 (C23), 58.9 (C20), 52.7 (C22), 20.8 (C13); HRMS (ESI) calculated for C23H32NO7 434.2173. Exp 434.2158 [M+H]+. . 4th compound 3e (120 mg, 0.36 mmol) is solubilized in a CH2Cl2 / MeOH mixture (2 / 3 mL) then cooled to 0°C. NaBH4 (15 mg, 0.40 mmol) is then added and after 15 minutes, the medium The reaction mixture is placed at room temperature for 4 hours. The reaction is then quenched with an IM HCl solution. The organic phase is washed with H2O (2x) then NaCl Sat, dried over Na2SO4, filtered, and evaporated. The crude product is then purified on a chromatographic column (SiO2, CH2Cl2 / acetone 80 / 20) to obtain the pure product as a beige solid (53 mg, 56%). R / (SiO2, CH2Cl2 / acetone, 80 / 20) = 0.17; *H NMR (300 MHz, CDC13, ô): 8.21 (s, 1H, H3), 7.67 (s, 1H, H5), 7.33 (d, 1H, J = 8.3 Hz, H12), 6.60 (dd, 1H, J3= 8.3 Hz, J4= 2.4 Hz, H11), 6.57 (d, 1H, J = 2.4 Hz, H9), 4.87 (d, 2H, J = 4.5 Hz, H17), 4.00 (s, 3H, H16), 3.87 (s, 3H, H13), 3.84 (s, 3H, H14), 3.42 (br, 1H, OH). O=S=O Compound 5a 18 5a 4a (126 mg, 0.439 mmol) is solubilized in dry CH2Cl2 (4 mL) and triethylamine (180 pL, 1.317 mmol) is added. After placing the medium at 0°C, mesyl chloride (50 pL, 0.702 mmol) is added and after 5 minutes, the reaction mixture is placed at room temperature for 30 minutes. The organic phase is washed with H2O, then dried over Na2SU4, filtered and evaporated to give a yellow oil used without further purification for the next step (137 mg, 85%). Rf (SiO2, CH2Cl2 / MeOH, 95 / 5) = 0.83; lH NMR (300 MHz, CDC13, 8): 8.08 (s, 1H, H3), 7.62 (s, 1H, H5), 7.18 (m, 1H, H12), 6.86-6.82 (m, 2H, Hn / H9), 5.47 (s, 2H, H17), 4.02 (s, 3H, H16), 3.85 (s, 3H, H14), 3.16 (s, 3H, H18), 2.30 (s, 3H, H13). o=s=o Compound 5b 24 5b 4b (282 mg, 0.67 mmol) is solubilized in CH₂Cl₂ (8 mL), and then triethylamine (280 pL, 2.02 mmol) is added. The mixture is placed at 0°C, and then mesyl chloride (80 pL, 1.08 mmol) is added. After 5 minutes, the reaction mixture is placed at room temperature for 30 minutes. The organic phase is washed with H₂O, and then the aqueous phases are re-extracted with CH₂Cl₂. The organic phases are collected, dried over Na₂SO₄, filtered, and evaporated under vacuum. The crude product is purified by column chromatography (SiO₂, CH₂Cl₂ / MeOH, 98 / 2 to 96 / 4) to obtain the pure compound as a pale yellow oil (304 mg, 91%). Rf (SiO2, CH2Cl2 / MeOH, 95 / 5) = 0.49 ; ’H NMR (500 MHz, CDC13, ô): 8.08 (d, 1H, J = 1.5 Hz, H3), 7.62 (d, 1H, J = 1.5 Hz, H5), 7.16 (d, 1H, J = 8.3 Hz, H12), 6.86 (d, 1H, J - 2.6 Hz, H9), 6.84 (dd, 1H, J3= 8.3 Hz, J4= 2.6 Hz, H11), 5.46 (s, 2H, H23), 4.17 (t, 2H, J= 4.7 Hz, H14), 4.01 (s, 3H, H22), 3.88 (t, 2H, J= 4.7 Hz, H15), 3.76-3.55 (m, 8H, H16 / H17 / H18 / H19), 3.38 (s, 3H, H20), 3.16 (s, 3H, H24), 2.28 (s, 3H, H13) ; 13C NMR (125 MHz, CDC13, Ô): 165.6 (C21), 159.5 (C10), 154.4 (C6), 152.4 (C4), 147.8 (C2), 136.9 (C8), 130.8 (C12), 130.5 (C7), 125.90 (C3), 125.88 (C5), 117.3 (C9), 112.6 (C11), 72.1 (C19), 71.2 (C23), 70.9 (C16), 70.8 (C17), 70.1 (C18), 69.9 (C15), 67.8 (C14), 59.2 (C20), 53.3 (C22), 38.3 (C24), 20.8 (C13) ; HRMS (ESI) calculée pour C23H32NO9S 498.1792. Exp 498.1779 [M+H]+. Composé 5c 19 5c 4c (56 mg, 0.18 mmol) is solubilized in CH2Cl2 (4 mL) and then triethylamine (75 pL, 0.53 mmol) is added. The mixture is placed at 0°C and mesyl chloride (21 pL, 0.27 mmol) is added. After 5 minutes, the reaction is placed at room temperature with stirring for 40 minutes. The organic phase is then washed with H2O and the aqueous phases are re-extracted with CH2Cl2. The organic phases are collected, dried over Na2SO4, filtered, and evaporated under vacuum. 1H NMR shows the quantitative yield of the pure product (70 mg), which is used without further purification for the next step. R / (SiO2, CH2Cl2 / MeOH, 95 / 5) = 0.64; 'H NMR (300 MHz, CDC13, ô): 8.04 (s, 1H, H3), 7.89 (s, 1H, NH), 7.59 (s, 1H, H45), 7.51 (s, 1H, H9), 7.46 (d, 1H, J= 8.3 Hz, H11), 7.16 (d, 1H, J= 8.3 Hz, H12), 5.43 (s, 2H, H18), 3.99 (s, 6H, H17), 3.14 (s, 3H, H19), 2.25 (s, 3H, H13), 2.19 (s, 3H, H15). O=S=O Compound 5d 24 5d 4d (123 mg, 0.28 mmol) is solubilized in CH2Q2 (5 mL) and then triethylamine (192 pL, 1.38 mmol) is added. The mixture is placed at 0°C and then mesyl chloride (53 pL, 0.68 mmol) is added. After 5 minutes, the reaction mixture is placed at room temperature for 30 minutes. The organic phase is washed with H2O and then the aqueous phases are re-extracted with CH2Cl2. The organic phases are collected, dried over Na2SO4, filtered and evaporated under vacuum. The product obtained is used without further purification for the next step. Rf (SiO2, CH2Cl2 / MeOH, 95 / 5) = 0.76; 'H NMR (300 MHz, CDC13, ô): 7.90 (s, 1H, H3), 7.45 (s, 1H, H5), 6.66 (s, 2H, H9), 5.44 (s, 2H, H23), 4.12 (t, 2H, J= 4.6 Hz, H14), 3.97 (s, 3H, H22), 3.83 (t, 2H, J= 4.6 Hz, H15), 3.74-3.51 (m, 8H, H16 / H17 / H18 / H19), 3.35 (s, 3H, H20), 3.13 (s, 3H, H24), 1.95 (s, 6H, H13); 13C NMR (75 MHz, CDCi3, ô): 165.3 (C21), 158.6 (C10), 154.7 (C6), 152.3 (C4), 148.0 (C2), 136.5 (C8 / !2), 130.4 (C7), 126.6 / 126.5 (C3 / C5), 113.9 (C9 / n), 71.9 (C19), 70.9 (C16), 70.7 (C17), 70.6 (C18), 69.7 (C15), 67.4 (C14), 59.0 (C20), 53.1 (C22), 38.1 (C23), . 5th grade compound 18 5th 4e (60 mg, 0.198 mmol) is solubilized in CH2Cl2 (3 mL) and triethylamine (83 pL, 0.593 mmol) is added. After placing the medium at 0°C, mesyl chloride (24 pL, 0.317 mmol) is added, and after 5 minutes, the reaction mixture is placed at room temperature for 30 minutes. The organic phase is washed with H2O, then dried over Na2SO4, filtered, and evaporated to give a yellow oil used without further purification for the next step (76 mg, quant.). Rf (SiO2, CH2Cl2 / MeOH, 95 / 5) = 0.76; 4H NMR (300 MHz, CDC13, ô): 8.29 (s, 1H, H3), 7.86 (s, 1H, H5), 7.35 (d, 1H, J = 8.4 Hz, H12), 6.61 (dd, 1H, J3= 8.4 Hz, J4= 2.3 Hz, H11), 6.57 (d, 1H, J - 2.3 Hz, H9), 5.46 (s, 2H, H17), 4.02 (s, 3H, H16), 3.87 (s, 3H, H13), 3.85 (s, 3H, H14), 3.14 (s, 3H, H18). Compound 6a 5 Triazacyclononane (27 mg, 0.115 mmol) and sodium carbonate (122 mg, 1.15 mmol) are solubilized in dry acetonitrile (7 mL). After 10 minutes at room temperature, 5a (130 mg, 0.356 mmol) is added and the reaction mixture is heated to 60°C with stirring for 24 h and then left at room temperature for 24 h. The mixture is then filtered through P3 sintered glass, rinsed with dry CH3CN, and the mother liquors are evaporated. The crude product is purified on a chromatographic column (Al2O3, CH2Cl2 / MeOH, 99:1 to 98:2) to obtain the pure product. d’un solide jaune pâle (81 mg, 75%). R / (A12O3, CH2Cl2 / MeOH, 95 / 5) = 0.58 ; *H NMR (500 MHz, CDC13, ô): 7.94 (s, 3H, H3), 7.68 (s, 3H, H5), 7.09 (d, 3H, J = 8.2 Hz, H12), 6.79-6.75 (m, 6H, H9 / Hn), 3.98 (s, 9H, H16), 3.97 (s, 6H, H17), 3.83 (s, 9H, H14), 2.92 (s, 12H, H18), 2.23 (s, 9H, H13) ; J3C NMR (125 MHz, CDCI3, Ô): 166.2 (C!5), 161.1 (C6), 159.9 (C10), 151.2 (C4), 15 147.3 (C2), 136.7 (C8), 131.1 (C7), 130.8 (C12), 127.0 (C5), 124.7 (C3), 116.3 (C9), 111.8 (C11), 64.8 (C17), 56.0 (C18), 55.4 (C14), 53.1 (C16), 20.9 (C13) ; HRMS (ESI) calculée pour C54H62N6O9 469.2284. Exp 469.2273 [M+2H]^. Triazacyclononane (93 mg, 0.39 mmol) and sodium carbonate (414 mg, 3.91 mmol) are dissolved in dry acetonitrile (24 mL) under argon. After 10 minutes at room temperature, 5b (289 mg, 1.21 mmol) is added, and the reaction mixture is heated at 60°C with stirring for 24 h, then at room temperature for another 24 h. The mixture is then filtered through a P3 frit, rinsed with dry CH3CN, and the mother liquors are evaporated. The crude product is purified by column chromatography (Al2O3, CH2Cl2 / MeOH, 99:1 to 95:5) to obtain the pure product in the form of a yellow oil (176 mg, 34%). 3H NMR (500 MHz, CDCI3, ô): 7.92 (d, 3H, J = 1.1 Hz, H3), 7.67 (s, 3H, H5), 7.07 (d, 1H, J = 8.4 Hz, H12), 6.81 (d, 1H, J = 2.4 Hz, H9), 6.77 10 (dd, 1H, J3= 8.4 Hz, J4= 2.4 Hz, H11), 4.14 (t, 6H, J- 4.6 Hz, H14), 3.96 (br s, 15H, H23 / H22), 3.86 (t, 6H, J= 4.6 Hz, H15), 3.74-3.53 (m, 24H, H16 / H17 / H18 / H19), 3.37 (s, 9H, H20), 2.91 (br s, 12H, H24), 2.21 (s, 9H, H13); 13C NMR (125 MHz, CDC13, ô): 166.2 (C2i), 161.0 (C6), 159.2 (Clü), 151.1 (C4), 147.3 (C2), 136.7 (C8), 131.3 (C7), 130.7 (C12), 127.0 (C5), 124.7 (C3), 117.0 (C9), 113.8 (C11), 72.1 (C19), 71.0 (C16), 70.8 (C17), 70.7 (C18), 69.8 (C15), 67.6 (C14), 15 64.7 (C23), 59.2 (C20), 55.9 (C24), 53.0 (C22), 20.8 (C13); HRMS (ESI) calculated for . C72H98N60ig 667.3463 Exp 667.34558 [M+2H]2+. Compound 6c 5c sec (70 mg, 0.18 mmol) is solubilized in dry acetonitrile (3 mL) and Na2CÛ3 (67 mg, 0.63 mmol) is added, along with triazacyclononane (13.6 mg, 0.06 mmol). After bubbling argon through the medium for 5 min, the reaction mixture is heated at 60°C for 5 days. The mixture is then filtered through a P3 sintered filter, rinsed with dry CH3CN, and the mother liquors are evaporated. The crude product is purified by chromatographic column (Al7O3, CJLCL / MeOH, 96 / 4) to obtain the pure product in the form of a white powder (47 mg, 81%). *H NMR (300 MHz, CDCI3, Ô): 8.94 (s, 3H, NH), 7.88 (s, 3H, H3), 7.54-7.45 (m, 9H, H5 / H9 / Hn), 7.07 (d, 1H, J- 8.3 Hz, H12), 3.98 (m, 15H, H18 / H17), 2.87 (s, 12H, H19), 2.25 (s, 9H, H13), 2.09 (s, 9H, H15); I3C NMR (75 MHz, MeOD, ô): 171.8 (C14), 166.9 (C16), 162.4 (C6), 152.8 (C4), 148.3 (C2), 140.7 (C10), 136.9 (C8), 134.9 (C7), 131.0 (C12), 128.6 (C3), 124.4 (C5), 123.0 (C9), 118.9 (CH), 65.0 (C18), 57.0 (C19), 53.3 (C17), 24.05 (C15), 20.9 (C13); HRMS (ESI) calculated for C57H64N9O9 1018.4822. Exp 1018.4782 [M+H]+;tR= 8.33 min (HPLC-MM-ACN method, H2O / CH3CN 85 / 15 to 0 / 100 in 16 min). 6d compound 6d Triazacyclononane (21.5 mg, 0.09 mmol) and sodium carbonate (95 mg, 0.90 mmol) are dissolved in dry acetonitrile (5 mL). After 5 minutes at room temperature, 5d (143 mg, 0.28 mmol) is added, argon bubbling is performed for 5 minutes, and the reaction mixture is heated at 60°C with stirring for 4 days. The mixture is then filtered through a P3 sintered filter, rinsed with dry CH3CN, and the mother liquors are evaporated. The crude product is purified by column chromatography (Al2O3, CH3Cl / MeOH, 100% to 97 / 3) to obtain the pure product as a white powder (82 mg, 66%). 'H NMR (300 MHz, CDCI3, ô): 7.77 (s, 3H, H3), 7.47 (s, 3H, H5), 6.64 (s, 6H, H9), 4.12 (t, 6H, J- 4.7 Hz, H14), 3.94 (s, 9H, H22), 3.91 (s, 6H, 10 H23), 3.84 (t, 6H, J= 4.7 Hz, H15), 3.74-3.52 (m, 24H, H16 / H17 / H18 / H19), 3.35 (s, 9H, H20), 2.82 (s, 12H, H24), 1.91 (s, 18H, H13) ; ,3C NMR (75 MHz, CDC13, ô): 166.0 (C21), 161.4 (C6), 158.3 (C10), 151.0 (C4), 147.5 (C2), 136.5 (Cs / I2), 131.3 (C7), 127.8 (C5), 125.3 (C3), 113.8 (C9 / l1), 72.0 (C19), 70.9 (C16), 70.7 (C17), 70.6 (C18), 69.8 (C15), 67.4 (C14), 64.6 (C23), 59.1 (C20), 55.6 (C24), 52.9 (C22), 21.0 (C13) ; HRMS (ESI) calculated for C75HI04N6O18 688.3698. Exp 15 688.3686 [M+2H]2+ ; tR= 13.09 min (HPLC-MM-ACN method, H2O / CH3CN 85 / 15 à 0 / 100 en 16 min). Composé 6e Triazacyclononane (15 mg, 0.064 mmol) and sodium carbonate (68 mg, 0.64 mmol) are solubilized in dry acetonitrile (4 mL). After 10 minutes at room temperature, 5e (76 mg, 0.198 mmol) is added and the reaction mixture is heated at 60°C with stirring for 24 h and then at room temperature for 5 days. The mixture is then filtered through P3 frit, rinsed with dry CH3CN and the mother liquors are evaporated. The crude product is purified on a chromatographic column (Al2O3, CH2Cl2 / MeOH, 100% to 97 / 3) to obtain the pure product as a white powder (41 mg, 65%). Rf (Al2O3, CH2Cl2 / MeOH, 95 / 5) = 0.54; 3H NMR (500 MHz, CDCI3, ô): 8.15 (s, 3H, H3), 7.92 (s, 3H, H5), 7.25 (d, 3H, J = 8.0 Hz, H12), 10 6.54-6.49 (m, 6H, H9 / Hu), 3.99 (s, 6H, H17), 3.97 (s, 9H, H16), 3.84 (s, 9H, H13), 3.75 (s, 9H, H14), 2.99 (s, 12H, H18); 13C NMR (125 MHz, CDCI3, ô): 166.5 (C15), 161.9 (C10), 160.9 (C6), 158.0 (C8), 147.8 (C4), 147.2 (C2), 131.4 (C12), 126.6 (C5), 124.4 (C3), 119.9 (C7), 105.3 (C9), 99.2 (C11), 65.0 (C17), 56.2 (C18), 55.7 (C13), 55.6 (C14), 53.0 (C16). Compound 7c 0 7c 6c (47 mg, 0.05 mmol) is dissolved in THF (3 mL), then NaOH IM (3 mL) is added until pH >12. The reaction mixture is stirred at room temperature for 24 h. LC-MS monitoring shows complete deprotection of the methyl esters. The product is used for complexation without further purification. HRMS (ESI) calculated 5 for C54H58N9O9 976.4352. Exp 976.4306 [M+H]+; tR = 6.24 min (HPLC-MM-ACN method, H2O / CH3CN 85 / 15 to 0 / 100 in 16 min). 6d (38.7 mg, 0.028 mmol) is dissolved in tetrahydrofuran (3 mL) and NaOH IM (3 mL) is added until pH 14 is reached. The reaction mixture is left to stir at room temperature. 10 for 6h30. Mass spectrometry monitoring shows the disappearance of the starting product. The mixture is used for the complexation step without further purification. HRMS (ESI) calculated for C75H98N6Oi8 667.3463. Exp 667.3461 [M+2H]2+ ; tR= 9.57 min (HPLC-ES1-ACN method, H2O / CH3CN 85 / 15 to 0 / 100 in 16 min). is added and the solution is stirred at room temperature for 1.5 hours. The pH is then adjusted The mixture is diluted to pH 4 with IM HCl (5 mL) and concentrated under vacuum. It is then diluted with MeOH and the pH adjusted to 8-9 using a saline Na₂CO₃ solution. Finally, TbCfI·0H₂O (84 mg, 0.224 mmol) is added, and the reaction mixture is stirred at room temperature for 72 h. The mixture is concentrated under reduced pressure, and the complex is precipitated with 5. Addition of TLO (2x) and centrifuged to obtain a white solid corresponding to the terbium(III) complex (79 mg, quant.). HRMS (ESI) calculated for Csi^NôOgTb 526.1558. Exp 526.1557 [M+2H]++. Shim for C5iH52N6O9Tb 1051.3044. Exp 1051.3011 [M+H]+. Ligand 6b (50 mg, 0.038 mmol) is solubilized in MeOH (5 mL) then NaOH IM 10 (2 mL) is added and the solution is left under stirring at room temperature for 1h. The pH is adjusted to pEI 3–4 with IM HCl, then to pH ~7 with Sat Na₂CO₃. TbCl₂·6H₂O (21 mg, 0.056 mmol) is added, and the solution is left at room temperature overnight. After concentration under reduced pressure, the mixture is purified by extractions / washes with DCM / H₂O, and the organic phase is evaporated to give the product as a pale yellow solid (55 mg, quant.). HRMS (ESI) calculated for C₆₉H₈₇N₆Na₂O₆TbCl₂O₆ is 746.2557.Exp 746.2565 [M+2Na]²⁺; tR = 9.65 min (HPLC-MM method, H₂O / CH₃CN 85 / 15 to 0 / 100 in 16 min). Complex Ia.3 The pH of the 7c solution is adjusted to pH 6 by adding an IM HCl solution, then TbCh.ôHzO is added at room temperature. The reaction mixture is left under stirring. for 48h, then the THF is evaporated under reduced pressure. The resulting precipitate is centrifuged 5 times in H2O (3x) to remove excess salts, and the white solid is then dried under vacuum (43 mg, 83%). HRMS (ESI) calculated for ^HssNgOgTb 1132.3371. Exp 1132.3323 [M+H]+, C54H54N9O9TbNa 1154.3190, Exp 1154.3133 [M+Na]+; tR = 6.67 min (HPLC-MM method) ACN, H2O / CH3CN 85 / 15 to 0 / 100 in 16 min). Complex Ia.4 10 The pH of solution 7d is adjusted to pH 6 with HCl IM then TbCl.ôH^O (12 mg, 0.031 mmol) is added and the solution is left under stirring at room temperature for 6 days. The THF is then evaporated and a CH2Cl2 / H2O extraction is performed. The organic phases are combined and evaporated to obtain the pure terbium(III) complex (38 mg, 90%). HRMS (ESI) calculated for C72H95N6Oi8Tb 745.2973. Exp 745.2966 [M+2H]2+, C72H94N6O8TbNa 756.2882. Exp 756.2882 [M+H+Na]2+ ; tR= 9.64 min (method HPLC-ESI-ACN-365, H2O / CH3CN 85 / 15 to 0 / 100 in 16 min). The ligand 6e (31 mg, 0.032 mmol) is dissolved in MeOH (5 mL), then NaOH IM (2 mL) is added, and the solution is stirred at room temperature for 2 h. The pH is then adjusted to pH 3–4 with HCl IM (2 mL) and then to pH 7 using a Na₂CO₃Sat solution. TbCl₂·6H₂O (18 mg, 0.047 mmol) is finally added, and the reaction mixture is stirred at room temperature for 14 h. The mixture is concentrated under reduced pressure, and the complex is precipitated with the addition of H₂O (2x) and centrifuged. The resulting solid is then precipitated again in Et₂O and 10 centrifuged to obtain a new white precipitate corresponding to the terbium(III) complex (30 mg, 85%). MS slab for C5iH53N60i2Tb 550.1488. Exp 550.1482 [M+2H]2+; slab for C5iH52N6NaO]2Tb 561.1392. Exp 561.1405 [M+H+Na]2+; slab for C5iH5iN6Na20i2Tb 572.1301. Exp 572.1323 [M+2Na]2+. Example 2: Dysprosium complex la.5 and Ia.6 Ligand 6b (50 mg, 0.038 mmol) is solubilized in MeOH (5 mL), then NaOH IM (2 mL) is added, and the solution is left under stirring at room temperature for 1 h. The pH is then adjusted to pH 3–4 with HCl IM, and then to pH ~7 with Na₂CO₃Sat. Dy(NO₃)₃·5H₂O (20 mg, 0.056 mmol) is added, and the solution is left at room temperature overnight. After concentration under reduced pressure, the mixture is purified by extractions / washes with DCM / H₂O, and the organic phase is evaporated to give the product as a pale yellow solid (50 mg, 92%). HRMS (ESI) calculated for C₆₉H₈₇DyN₆Na₂O₇²⁺: 748.7577. Exp: 748.7576 [M+2Na]²⁺. 6d (9 mg, 0.007 mmol) is dissolved in tetrahydrofuran (2 mL) and NaOH IM (2 mL) is added until pH 14 is reached. The reaction mixture is left under stirring at room temperature for 24 h. The pH of the solution is then adjusted to pH 6 with HCl IM, and DyCl3·6H2O (3.2 mg, 0.008 mmol) is added. The solution is left under stirring at room temperature for 3 days. The THF is then evaporated, and a CH2Cl2 / H2O extraction is performed. The organic phases are collected and evaporated to obtain the pure dysprosium(III) complex (8 mg, 77%). HRMS (ESI) calculated for C72H93N6O8DyNa2 769.7811. Exp 769.7818 [M+2Na]2+, C72H93N6Oi8DyNa3 520.8505. Exp 520.8521 [M+3Na]3+ ; tR= 9.57 min (HPLC-ESI-ACN-365 method, H2O / CH3CN 85 / 15 to 0 / 100 in 16 min). B. EVALUATION OF COMPLEXES Example 3: Spectroscopic properties The properties of complexes Ia.l-Ia.6 and 8 were evaluated in methanol (Table 1) and in water (Table 2). These properties were compared to those of complex 9, described in Dalton Trans. 2015, 44, 4918, and with the following structure: MeO MeO 9 Photophysical measurements of the complexes show that the terbium Ia.l-Id.4 complexes according to the invention have intense absorption in methanol (Table 1). Furthermore, these complexes exhibit very high quantum yields, significantly exceeding that of complex 9 outside the scope of this invention, which has no substituents other than hydrogens alpha to the pyridine-phenyl bond (positions R2 and R3). The strong limitation, or even suppression, of internal rotation around the pyridine-phenyl bond thus allows for a significant improvement in quantum yield. Consequently, the brightness of the complexes according to the invention is superior to that of complex 9. Different electron-donating groups were tested: methoxy, PEG, or amide. Less favorable results were observed with an amide group (complex Ia.3), due to a lower quantum yield, but the results were still satisfactory in terms of brightness. On the other hand, the presence of a second electron-donating group of the methoxy type on the phenyl group (complex 8) causes a drop in quantum yield, compared to a methyl group (complex la.l). The introduction of a second methyl group has a major effect on the molar extinction coefficient, with a decrease of more than 50%. Thus, despite excellent quantum yield, Ia.4 exhibits a lower brightness at 330 nm than its counterpart Ia.2. The spectroscopic properties of complexes Ia.2 and Ia.4 are conserved in water (Table 2). The brightness of the Ia.2 complex is very high, and comparable to that of the TbLumi-4 complex, from the work of the group of K. Raymond (J. Xu et al. J. Am. Chem. Soc. 2011, 133, 19900-19910), which is among the best complexes currently on the market (O = 59%, Σ of the order of 25000 L.moT1. cm'1 and a brightness around 15000 L.moL1. cm'1). Figure 1 shows the absorption and emission spectrum of the Ia.4 complex in water at room temperature. Table 1 - Photophysical properties of complexes Ia.1-Ia.6, 8 and 9 measured in methanol Complex X max (nm) E (L.moF1. cm'1) © (%) T (ms) B at X max (L.mol'1. cm'1) B at 330 nm (L.moF1. cm' ') la.l 306 21000 74 1.33 15500 nd Ia.2 305 34800 74 1.32 26000 10000 Ia.3 305 34800 35 0.79 12000 5600 Ia.4 274 302 19000 13000 65 1.59 12000 8500 3400 8 (outside the scope of this invention) 325 51000 13 0.22 6600 6500 Ia.5 306 nd 2.8 0.021 ndnd Ia.6 301 nd 2.6 0.020 ndnd 9 (excluding invention) 308 33000 12 nd 3900 nd Table 2 - Photophysical properties of complexes Ia.2 and Ia.4-Ia.6 measured in water complex X max (nm) E (L.moF1. cm'1) O (%) T (ms) B at 11800 66 1.46 14000 7800 2500 Ia.5 302 nd 1.8 0.017 ndnd Ia.6 300 nd 2.5 0.028 ndnd 5. Dysprosium complexes Ia.5 and Ia.6 also exhibit properties remarkable photophysical properties in methanol and water. These complexes are among the brightest reported in the literature. Figure 2 shows the absorption and emission spectrum of complex Ia.6 in a methanol / ethanol mixture at room temperature. 10 Example 4: Cell imaging The Ia.2 complex has shown significant potential in one- and two-photon fluorescence microscopy imaging. Figure 3 shows images of cells fixed to PFA and labeled with the Ia.2 complex (concentration = 15 mol·L⁻¹, two-photon excitation, Xₑₓ = 720 nm), clearly demonstrating its internalization within the cell. The emission spectra confirm the presence of Ia.2 inside the cell and not in the buffer. Figure 4 shows images of T24 cells fixed to PFA and labeled with complex Ia.5 (concentration = 10⁵ mol.L⁻¹, two-photon excitation, Xex = 720 nm). As with complex Ia.2, the internalization of complex Ia.5 into the cell is confirmed by the emission spectra.

Claims

DEMANDS 1. Lanthanide complex comprising a chelating agent, formed of a macrocycle or a ligand, complexing a lanthanide ion Ln'. characterized in that the lanthanide is selected from terbium and dysprosium and in that the chelating agent comprises at least one group (B) of the following structure: in which: -RI represents a hydrogen, an -R6 group, or an electrodominant group -El, -R2 represents a hydrogen, an -R7 group, or an electron-donating group -E2, -R3 represents a hydrogen, an -R8 group, or an electron-donating group -E3, -R4 and -R5, whether identical or different, independently represent a hydrogen or a group -R9 or -OR9, -El, -E2 and -E3 are chosen independently of each other from the groups -OR10, -SR10, -NH(CO)R10, -NH(CO)NR10R'10, -NH(CS)NR10 R'10 and -NH(CS)NHR10, -R6, -R7, -R8, -R9, -RIO and R'10, whether identical or different, independently represent an (C1-C6) alkyl group, possibly substituted by an -XI group or a -Y group, -XI is a reactive group, -Y is a water-solubilizing group, it being understood that: or at least one of the substituents -R2 and -R3 is not a hydrogen, o only one of the groups -RI, -R2 and -R3 represents an electron-donating group, o said chelating agent comprises at most one reactive group.

2. Lanthanide complex according to claim 1 selected from the lanthanide complexes of formula (I): Chromel Chrom3 (I) in which: - Ln is a lanthanide chosen from Tb and Dy, -A- represents -CH2- or -CH(La-X2)-, 5 - -La- is a covalent bond, or a linear or branched (C1-C20) alkylene group, possibly containing one or more double or triple bonds, and possibly substituted by one to three -SO3H groups, or -La- is a (C5-C8) cycloalkylene group or a (C6-C14) arylene group, -X2 is a reactive group, 10 - -Chroml, -Chrom2 and -Chrom3 are identical or different and chosen independently of each other among the formula groups (Bl): -RI, -R2, -R3, -R4, and -R5 are as defined in claim 1, -Z- represents -C- or ~P(Rz)-, and 15 - -Rz represents a phenyl, benzyl, methyl, ethyl, propyl, n-butyl group, sec-butyl, iso-butyl or tert-butyl, and preferably a phenyl or methyl group, it being understood that if -A- is -CH(La-X2)-, then none of the -Chroml, -Chrom2 and -Chrom3 groups of formula (Bl) includes a -XI group.

3. Lanthanide complex according to claim 2, characterized in that -A- represents 20 -CH?- and -Z- represents -C-.

4. Lanthanide complex according to claim 2 or 3, characterized in that -Chrom1, -Chrom2 and -Chrom3 are identical, preferably with: -RI representing an electron-donating group -El as defined in claim 1, in particular chosen from the groups -OR10 and -NH(CO)R10, -RIO being such as 25 as defined in claim 1, preferably -RI represents -OMe or -OPEG, -R2 being a group -R7 as defined in claim 1, preferably -Me, and -R3 being a group -R8 as defined in claim 1, and in particular -Me, or, preferably, a hydrogen.

5. Lanthanide complex according to claim 2 or 3, wherein: 5 - -A- represents -CH2-, -Chroml and -Chrom2 are identical and have the (B2) structure: R11 with -RI1, -R21, -R31, -R41, -R51 and -Z1- as defined respectively for RI, -R2, -R3, -R4, -R5 and -Z- in any one of claims 1 to 3, it being understood that 10 Chroml and -Chrom2 do not include a reactive group, and -Chrom3 is different from -Chroml and -Chrom2 and has the structure (B3): R12 with -RI2, -R22, -R32, -R42, -R52 and -Z2- as defined respectively for -RI, -R2, -R3, -R4, -R5 and -Z- in any one of claims 1 to 3, 15, it being understood that -Chrom3 includes a reactive group, and preferably: -RI1 is an -O(C1-C6) alkyl group substituted by a -Y group, -RI2 is an -O(C1-C6) alkyl group substituted by an -XI group, - -R21 and -R22, whether identical or different, represent a group -R7 as defined in the 20 claim 1, and are preferably identical and preferably represent, an alkyl (C1-C6) group, and in particular -Me, and -R31, -R32, -R41, -R42, -R51, and -R52 are hydrogens.

6. Lanthanide complex according to claim 1 selected from the lanthanide complexes of formula (II): Rn^Ghrom4^ ri 2 -OOC^N Nf^COO- I ln3+ ] Chrom6 Chromô -OOC7 XCOO- in which: - Ln is a lanthanide chosen from Tb and Dy, - -Rll and -R12, identical or different, are chosen independently of each other from -XI, -Y, a hydrogen atom or an (C1-C6) alkyl group, -Chrom4-, -Chrom5-, and -Chromé-, identical or different, independently represent a group of formula (B), as defined in claim 1.

7. Lanthanide complex according to claim 6, characterized in that -Chrom4-, -Chrom5- and -Chromated- are identical, preferably with: -RI representing an electron-donating group -El as defined in claim 1, in particular chosen from the groups -OR10 and -NH(CO)R10, -RIO being as defined in claim 1, preferably -RI representing -OMe or -OPEG, - -R2 being a group -R7 as defined in claim 1, preferably -Me, and -R3 being a group -R8 as defined in claim 1, and represents in particular -Me, or, preferably, is a hydrogen.

8. Lanthanide complex according to claim 6, characterized in that: ® or: -Chrom5- and -Chromé- are identical and have the same structure (B 10): with -RI3, -R23, -R33, -R43, and -R53 as defined respectively for -RI, -R2, -R3, -R4, and -R5 in claim 1, it being understood that -Chrom5- and -Chromé- do not include a reactive group, -Chrom4- is different from -Chrom5- and -Chromé- and has the structure (Bll): with -RI4, -R24, -R34, -R44 and -R54 as defined respectively for -RI, -R2, -R3, -R4 and -R5 in claim 6, it being understood that -Chrom4- comprises a reactive group -XI, • either : 10 - -Chrom4- and -Chromé- are identical and have the same structure (B 12): with -RI5, -R25, -R35, -R45 and -R55 as defined respectively for -RI, -R2, -R3, -R4 and -R5 in claim 1, it being understood that -Chrom4- and -Chromé- do not comprise a reactive group, and 15 - -Chrom5- is different from -Chrom4- and -Chromé- and has a structure of (B13): with -RI6, -R26, -R36, -R46 and -R56 as defined respectively for -RI, -R2, -R3, -R4 and -R5 in claim 1, it being understood that -Chrom5- comprises a reactive group -XI.

9. Lanthanide complex according to any one of claims 1 to 8, characterized in that -R4 and -R5 represent hydrogens.

10. Lanthanide complex according to any one of claims 1 to 9, characterized in that it comprises a single reactive group enabling its covalent coupling to a biomolecule, said reactive group being preferably selected from -COOH, -NH2, an acrylamide, an activated amine, an activated ester, an aldehyde, an alkyl halide, an anhydride, aniline, an azide, an aziridine, a carboxylic acid, a diazoalkane, a haloacetamide, a halotriazine, a hydrazine, an imido ester, an isocyanate, an isothiocyanate, a maleimide, a sulfonyl halide, a thiol, a ketone, an acid halide, a hydroxysuccinimidyl ester, a succinimidyl ester, a hydroxysulfosuccinimidyl ester, an azidonitrophenyl, a azidophenyl, a 3-(2-pyridyl dithio)-propionamide, a glyoxal, a triazine, an acetylenic group, and the formula groups: O ^n^-(ch2)ncs O / 'N}4CH2)NH2 ri vw in which w is an integer in the range from 0 to 8 and v is equal to 0 or 1, and Ar is a saturated or unsaturated 5- or 6-membered heterocycle comprising 1 to 3 heteroatoms, possibly substituted by a halogen atom; reactive groups chosen from -COOH, -NH2, succinimidyl esters, haloacetamides, azides, hydrazines, isocyanates and maleimides being preferred.

11. Lanthanide complex according to any one of claims 1 to 10, characterized in that the group Y is selected from -SO3', -COO', the sulfobétaine groups and -O-[(CH2)2-O]m-CH3, m being an integer from 1 to 10, preferably m=3.

12. Lanthanide complex according to any one of claims 1 to 11, characterized in that Ln is terbium.

13. Chelating agent of formula (III): Chroml' IN—Chrome2 Chrom3' (III) with -Chroml', -Chrom2', and -Chrom3', identical or different, and chosen independently of each other from the formula groups (B5): with RI, R2, R3, R4 and R5 as defined in claim 1, and -R13 an acid-protecting group such as an alkyl, and in particular a methyl, or in the form of a salt.

14. Chelating agent of formula (IV): Chrom6 Chrom5 R13COO COOR13 ^j^) with -Chrom4-, -Chrom5- and -Chromé- as defined in any one of claims 6 to 8, and -R13 an acid-protecting group such as an alkyl, and in particular a methyl, or in the form of a salt.

15. A method for detecting a biomolecule comprising detecting the luminescence of a conjugate of said biomolecule with a luminescent complex according to any one of the claims 1 to 12, comprising a reactive group, and obtained by coupling said biomolecule with said luminescent complex on its reactive group.

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