Metal complex compounds as photosensitizers for photodynamic therapy.

Abstract

Electroneutral complex compounds of formula (I), including any stereoisomers or E / Z isomers, wherein M is Fe 2+ , Ru 2+ , Os 2+ , Co 3+ , Rh 2+ , Rh 3+ , Ir 3+ , Ni 2+ , Pd 2+ , Pt 2+ or Pt 4+ , especially Ru 2+ where T and T' are H or (C1-C3)-alkyl, Lig is a bidentate ligand, particularly 2,2'-bipyridine (bpy) or 4,7-diphenyl-1,10-phenanthroline (BPhen), and A is an anion derived from a pharmaceutically acceptable acid, is useful in human therapy, particularly as a photosensitizer (PS) in photodynamic therapy (PDT) of cancer, certain skin conditions, and fungal or microbial infections. The complex compounds are substantially nontoxic under dark conditions at therapeutic doses and exhibit good to excellent photoactivity under hypoxic conditions. The complex compounds are useful for PDT treatment of deep hypoxic tumors using light within the phototherapeutic range from far-infrared to near-infrared. TIFF2026508498000052.tif87170

Description

[Technical Field]

[0001] The present invention relates to novel metal complex compounds useful as photosensitizers (PS) for photodynamic therapy (PDT), and to novel intermediate ligand compounds useful for preparing such complex compounds. [Background technology]

[0002] Photodynamic therapy, in various forms, has been around for over 100 years. In recent years, PDT has emerged as a promising medical intervention for treating certain types of cancer that cannot be treated by conventional methods. PDT has also been used to improve some skin conditions (e.g., acne or port-wine stains), some fungal and microbial infections, and some cases of age-related macular degeneration (see Non-Patent Document 1 and references therein).

[0003] PDT is a well-established noninvasive modality for destroying tumors and / or tumor blood vessels based on the combination of three components: a photosensitizer (PS), light of an appropriate wavelength (λ), and oxygen. PDT is performed in a two-step procedure involving the local or systemic administration of a nontoxic amount of PS followed by photoactivation. Compared with other conventional treatment protocols, PDT offers several advantages in the treatment of cancer due to its noninvasive nature and the spatiotemporal selectivity afforded by light. In fact, a key attribute of PDT is that toxicity is limited to the region where the three components (PS, light, and oxygen) overlap in space and time.

[0004] The PDT effect results from the excitation of PS to generate a singlet excited state, which undergoes intersystem crossing to form a reactive triplet state. The triplet state then converts to cytotoxic singlet oxygen (COO) via type II energy transfer (the major pathway). 1 O2) or participate in type I electron transfer reactions to generate other reactive oxygen species such as superoxide radicals or hydroxyl radicals.

[0005] Photofrin™ and protoporphyrin IX (PpIX), both of which contain a tetrapyrrole ring system, are approved worldwide for the treatment of certain cancers (see page 24 of Non-Patent Document 1). While these two compounds have become the standard for PDT, there is ongoing interest in developing novel PSs that overcome some of the limitations of existing PSs for clinical use.

[0006] Water-soluble PS prepared with high purity by simple synthesis is highly desirable. Certain metal complex compounds hold great promise in this regard. Several metal ion-containing PSs have advanced to clinical trials, with the most promising being padeliporfin (also known as TOOKAD™ Soluble and WST11) and TLD-1433 (see page 25 of Non-Patent Document 1). Padeliporfin is a tetrapyrrole Pd(II) complex compound used as a PS in vascular-targeted PDT to treat low-risk prostate cancer. TLD-1433 is the first Ru(II) complex compound used as a PS in a PDT clinical trial to treat non-muscle-invasive bladder cancer. TLD-1433 is a thiophene-containing complex compound disclosed in a series of patent documents by SA McFarland with priority dates of 2012. This patent family includes several issued patents, such as US Pat. No. 5,629,992 and US Pat. No. 5,629,992, both of which claim PS, which are thiophene-containing complex compounds of several metals (Mn, Mo, Re, Fe, Ru, Os, Co, Rh, Ir, Ni, Pt, and Cu).

[0007] PDT is an oxygen-dependent process, which reduces its efficiency in treating deeply hypoxic tumors (i.e., tumor microenvironments lacking sufficient oxygen supply at the tissue level). On the other hand, although excitation with longer wavelengths of light is generally preferred to enhance tissue penetration, some known PSs generally tend to be less effective at promoting the generation of type II reactive oxygen species (i.e., singlet oxygen). There is ongoing interest in developing novel PSs based on metal complexes for PDT that address some of the limitations of known PSs, particularly those with operability in the far-infrared and near-infrared (NIR) regions of the electromagnetic spectrum and under hypoxic conditions. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] European Patent No. 2854948 [Patent Document 2] U.S. Patent No. 9,676,806 [Non-patent literature]

[0009] [Non-Patent Document 1] SA McFarland et al.; "Metal-based photosensitizers for photodynamic therapy: the future of multimodal oncology?"; Current Opinion in Chemical Biology, 2020, vol. 56, pp. 23-27 Summary of the Invention

[0010] One aspect of the present disclosure relates to providing compounds of Formula I or Formula IV, including stereoisomers and E / Z isomers. [ka] TIFF2026508498000003.tif137170 (in the formula, M is Fe 2+ , Ru 2+ , Os 2+ , Co 3+ , Rh 2+ , Rh 3+ , Ir 3+ , Ni 2+ , Pd 2+ , Pt 2+ , and Pt 4+ is a metal cation selected from the group consisting of y=1, 2 or 3, and z=0, 1 or 2, provided that M is Ni 2+ , Pd 2+ or Pt 2+ When y+z=2, M is Fe 2+ , Ru 2+ , Os 2+ , Co 3+ , Rh 2+ , Rh 3+ , Ir 3+ , or Pt 4+ If , then y+z=3, m=0, 1, 2, 3 or 4; the groups T and T′ at each occurrence are each independently selected from the group consisting of H and (C1-C3)-alkyl; The groups P3, P5, P6, Q3, Q4, Q5, Q6, R3 (the group R3 is not included in formula IV, so the definition of R3 applies only to formula I), R5, R6, and R8 each in each occurrence are respectively H, (C1-C3)-alkyl, (C3-C6)-cycloalkyl, CH=CHR, CF3, CHF2, CH2F, CF2CF3, F, Cl, Br, I, OR, C(=O)OR, O(C=O)R, C(=O)NR, NRC(=O)R', N RR' is a group independently selected from the group consisting of phenyl, monosubstituted phenyl, disubstituted phenyl, and trisubstituted phenyl, wherein the substituents on the phenyl ring are attached at any of the possible substitution positions and are independently selected from the group consisting of F, Cl, Br, I, NO2, (C1-C3)-alkyl, OH, O[(C1-C3)-alkyl], NH2, NH[(C1-C3)-alkyl], and N[(C1-C3)-alkyl]2; The group R7 is H, NO2, OR, NRR', N(CH2COOH)2, N(CH2CH2SO3H)2, N(CH2CONHCH2CH2NMe2)2, or the heterocyclic groups 1-aziridinyl, 1-azetidinyl, 1-pyrrolidinyl, 1-piperidinyl, 4-morpholinyl, or 1-piperazinyl, respectively, unsubstituted, mono-(R)-substituted, di-(R,R')-substituted, or tri-(R,R',R'')-substituted; alternatively, R6, R7, and R8 together form a bicyclic system of the following formula: 10 , V 11 and V 12 are each a group independently selected from the group consisting of H and (C1-C3)-alkyl, [ka] The group R4 is H, (C1-C3)-alkyl, (C3-C6)-cycloalkyl, CF3, CHF2, CH2F, CF2CF3, F, Cl, Br, I, OR, C(=O)OR, O(C=O)R, C(=O)NR, NRC(=O)R', NRR', phenyl, mono-(R)-substituted phenyl, di-(R,R')-substituted phenyl, tri-(R,R',R'')-substituted phenyl, or the group CE1E2 E3, where E1 and E2 are groups independently selected from the group consisting of H, F, Cl, Br, I, OR, NRR', NO2, (C1-C3)-alkyl, phenyl, and mono-(R)-substituted phenyl, E3 is OH, O[(C1-C3)-alkyl], or a group of the following formula: where p=0, 1, 2, 3, or 4, and D1 and D2, in each occurrence, are H, O[CH2] q -X, S[CH2] q -X, and N[[CH2] q -X]2, where q=1 or 2 and X=F, Cl, Br or I; [ka] or a group of the following formula (wherein p=1, 2, 3 or 4, and D3 is a group independently selected from the group consisting of H, H(C=O), [(C1-C3)-alkyl](C=O), (OH)2P(=O)O-CH2-O(C=O), and (PhO)(OH)P(=O)): [ka] and Lig, at each occurrence, is a bidentate ligand independently selected from the group consisting of: Lig1, Lig2, Lig3, Lig4, Lig5, Lig6, Lig7, Lig8, Lig9, Lig10, Lig11, Lig12, and Lig13, wherein G is a N atom or a H atom in the formation of a carbon-metal bond. + is one of the C atoms with one negative charge formed in situ from C-H when R is lost. 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , R 20 and R 21 each occurrence is a radical independently selected from the group consisting of H, (C1-C3)-alkyl, phenyl, and mono-, di-, and tri-substituted phenyl, wherein the substituents attached to the phenyl ring are independently selected from F, Cl, Br, I, OR, and NRR'; [ka] TIFF2026508498000008.tif225170A is an anion derived from a pharmaceutically acceptable acid, n is an integer or fraction such that the compound of formula I is electrically neutral; wherein R, R', and R'' at each occurrence above are each independently selected from the group consisting of H, F, Cl, Br, I, NO2, (C1-C3)-alkyl, OH, O[(C1-C3)-alkyl], NH2, NH[(C1-C3)-alkyl], and N[(C1-C3)-alkyl]2.

[0011] As used herein, the term "stereoisomer" is a general term referring to all isomers that differ only in the spatial arrangement of atoms in individual molecules, and includes enantiomers and diastereomers. If a compound has a chiral center, it can exist in various stereoisomeric forms, such as enantiomeric or diastereomeric forms. Thus, any given compound referred to herein is intended to represent either the racemate, one or more enantiomeric forms, and one or more diastereomeric forms. All stereoisomers, including enantiomers and diastereoisomers of the compounds referred to herein, and mixtures thereof (including racemic mixtures, enantiomerically enriched mixtures, and diastereomerically enriched mixtures), are considered within the scope of the present disclosure. The present disclosure also contemplates all possible E / Z isomers and mixtures thereof.

[0012] It is to be noted that in a preferred embodiment of the present invention, formula IV can be obtained from formula I when, in formula I, m≧1 (e.g., 1, 2, 3 or more), and group R3 which is CH2 forms a six-membered ring with the T′ group which is closest to the coumarin skeleton, provided that T′=CH2. It is further to be noted that in another preferred embodiment of the present invention, formula V can be obtained from formula II when, in formula II, m≧1 (e.g., 1, 2, 3 or more), and group R3 which is CH2 forms a six-membered ring with the T′ group which is closest to the coumarin skeleton, provided that T′=CH2.

[0013] Generally, the complex compounds of formula I or formula IV can be obtained by reacting the corresponding intermediate compounds of formula II or formula V (respectively), including the corresponding stereoisomers or E / Z isomers, with the corresponding reactant compounds of formula III in a solvent or solvent mixture, where m, n, y, z, P3, P5, P6, Q3, Q4, Q5, Q6, R3, R4, R5, R6, R7, R8, T, T', M, Lig, and A are as defined above. Because the intermediate compounds of formula II or formula V are novel, another aspect of the present disclosure relates to providing these compounds that are useful in the preparation of the complex compounds of formula I or formula IV. [ka]

[0014] Generally, the reactant compound of formula III and the intermediate compound of formula II or V are reacted in a molar ratio substantially corresponding to the stoichiometry determined by the values ​​of z and y, although in some cases it may be advisable to use an excess of III to improve the yield. In some cases, the intermediate compound is obtained using a different anion A', which is finally subjected to an anion exchange reaction (e.g., using an ion exchange resin).

[0015] As shown in the results presented in the Detailed Description section, the metal complex compounds of Formula I or Formula IV are useful as PS in anti-cancer PDT. For example, the complex compounds of Formula Ia and Formula Ic show excellent phototoxicity results against the CT-26 mouse colon cancer cell line.

[0016] Therefore, another aspect of the present disclosure relates to providing a pharmaceutical composition comprising a therapeutically effective amount of a complex compound of Formula I or Formula IV together with a suitable amount of an excipient, carrier or vehicle.

[0017] The complex compounds of formula I are substantially non-toxic under dark conditions at therapeutic doses. For example, both complex compounds Ia and Ic are non-toxic under dark conditions, and IC 50 The IC values ​​of both compounds are above 250 μM, indicating strong cytotoxicity after irradiation with monochromatic visible light. As shown in Table 1, the IC values ​​of both compounds were50 The values ​​were found in the low nanomolar range (8.2 nM at 540 nm for compound Ia and 7.4 nM at 645 nm for compound Ic), with very good PI values ​​(>30487 and >33783, respectively). For comparison, the reported results for TLD-1433, a PS in clinical development, are as follows: IC of TLD-1433 in SK-MEL-28 melanoma cells 50 The values ​​are in the low micromolar range (2.3 μM) after red light (625 nm) irradiation, indicating a moderate PI value (see S. Monro et al.; "Transition metal complexes and photodynamic therapy from a tumor-centered approach: challenges, opportunities, and highlights from the development of TLD-1433"; Chem. Rev. 2019, vol. 119, pp. 797-828).

[0018] Compounds of formula Ic inhibited the CT-26 cancer cell line (IC 50 =0.76 μM, PI>329) and its human analogue HT-29 cancer cell line (IC 50 =0.35 μM, PI>714), these results are better than those corresponding to the control drug, protoporphyrin IX. As shown in the results in Table 6, similar results are obtained with other human cancer cell lines (A-549 and A-2780). Similarly, as shown in Tables 5 and 8, the compounds of Formula Im, Formula In, Formula Io, and Formula IVa were found to be phototoxic upon irradiation with NIR light (740 nm and 770 nm) in the CT-26 cancer cell line (e.g., IC of Im at 740 nm). 50 =0.17μM, PI>588).

[0019] The complex compounds of Formula I or Formula IV exhibit good to excellent photoactivity under hypoxic conditions. For example, Compound Ia and Compound Ic exhibit excellent photoactivity under hypoxic conditions, with Compound Ia being particularly toxic when irradiated at 540 nm and exhibiting an excellent PI of greater than 7143. Compound Ic exhibits a PI value of about 2900 to about 3300 in the light irradiation range of 540 nm to 670 nm. Compound Im exhibits a submicromolar IC when irradiated with NIR light (740 nm) under hypoxic conditions. 50 These results indicate that Compound I or Compound IV is useful for PDT treatment of deep hypoxic tumors using light within the phototherapeutic range from far infrared to near infrared.

[0020] Aspects of the present disclosure relate to complex compounds of Formula I or Formula IV for use in human treatment, particularly as photosensitizers in photodynamic therapy of human conditions, more particularly, cancer, certain skin conditions, fungal infections, or microbial infections. These aspects also relate to processes for the preparation of medicaments for photodynamic therapy in humans of cancer, certain skin conditions, fungal infections, or microbial infections, comprising the use of complex compounds of Formula I or Formula IV, preferably in the form of nanoformulations. In other words, complex compounds of Formula I or Formula IV can be useful in methods of photodynamic therapy for the treatment of cancer, certain skin conditions, fungal infections, or microbial infections in humans.

[0021] Throughout this disclosure and claims, the word "comprise" and variations thereof are not intended to exclude other technical features, additives, elements, or steps. Furthermore, the word "comprise" also encompasses the term "consisting of." Additional objects, advantages, and features will become apparent to those skilled in the art upon examination of this disclosure or may be learned by practice of the disclosure. The following examples are offered by way of illustration and not by way of limitation. [Brief explanation of the drawings]

[0022] [Figure 1] Top: Growth of 3D CT-26 multicellular tumor spheroids (MCTS) monitored over 7 days compared to initial size in the presence of increasing concentrations of Ic. Data are shown as the mean ± SD from three independent measurements. Control: Untreated MCTS, irradiated with 645 nm light (2.50 mW / cm², 9.00 J / cm²) for 1 hour. Treated MCTS were incubated with Ic for 36 hours and then irradiated with 645 nm light (2.50 mW / cm², 9.00 J / cm²) for 1 hour. Bottom: Photomicrographs of 3D CT-26 MCTS taken over 7 days (scale bar = 1000 µm). [Figure 2] Figure 1 shows the cellular uptake of complex compound Ic. Single confocal planes of HeLa cells incubated with compound (10 μM) for 30 min at 37°C were imaged at t=0 and 2 min after the first observation. Excitation was performed using three different laser lines (405 nm, 458 nm, and 514 nm). White arrows point to mitochondria, and white arrowheads point to vesicular staining. Black arrowheads in the right column point to cell blebs. Scale bar: 20 μm. LUT for compound images: Fire. Left and right columns: merged compound and brightfield images. [Figure 3] Figure 1: Colocalization study of Ic using Mitoview 650. Single confocal plane of HeLa cells incubated with compound (10 μM, green) and Mitoview (0.1 μM, red). Left: Overlay of the two stainings. Center: Ic' signal. Right: Mitoview signal. White arrows and white arrowheads indicate positive and negative (vesicular staining in compound images) colocalization, respectively. Scale bar: 20 μm. [Figure 4] Mean plasma Ic concentrations in adult male albino Swiss CD1 mice at different times after IP administration of 5 mg / kg (left) and pharmacokinetic parameters of Ic in mouse plasma (right). [Figure 5]Figure 1 shows body weight (g) (left) and food consumption (g) per animal (right) during the 5-day experimental protocol. CD1 mice were treated intraperitoneally with vehicle and complexed compound Ic (10 mg / kg or 30 mg / kg) on ​​day 1 and sacrificed on day 5. Results are expressed as mean ± SD (n = 3 males, n = 3 females). [Figure 6] Figure 1 shows the organ weight (mg) / body weight (g) ratios for animals treated intraperitoneally with vehicle and complex compound Ic (10 mg / kg or 30 mg / kg) on ​​day 1 and sacrificed on day 5. Results are expressed as mean ± SD (n = 3 males, n = 3 females). *p < 0.05 vs. vehicle group for males or females (1-way ANOVA and Bonferroni post-hoc test). [Figure 7] Figure 1 shows an in vivo PDT efficacy study of Ic (after IT administration) in female BALB / c mice bearing subcutaneous CT-26 syngeneic colon tumors. Irradiation was performed with 660 nm LED light (15 min, 100 mW / cm2). Tumor growth inhibition curves (left) and average tumor weights (right) of treated tumor-bearing mice (D: dark group, L: irradiated group, xn: number of irradiations). Data are presented as the mean ± SEM of five mice treated in parallel (n = 5). Mean tumor weight data were analyzed using one-way ANOVA followed by Bonferroni's multiple comparison test (asterisks: *p < 0.05, **p < 0.001). DETAILED DESCRIPTION OF THE INVENTION

[0023] In certain embodiments of the complex compound of Formula I or Formula IV, the anion A is Cl - , Br - , PF6 - , PF4 - , BF4 - , ClO4 - , CF3SO3 - , SO4 2- , CF3COO - , acetic acid, formic acid, and oxalic acid. In certain embodiments, A is Cl - is.

[0024] In certain embodiments of the complex compound of Formula I or Formula IV, y=1, z=2, and the metal cation M is Fe 2+ , Ru 2+ , Os 2+ , Co 3+ , Rh 2+ , Rh 3+ , Ir 3+ , or Pt 4+ In certain embodiments, the metal cation M is Ru 2+ is.

[0025] In certain embodiments of the complex compound of formula I or formula IV, the bidentate ligand Lig is Lig1 or Lig2 and the group R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , R 20 and R 21 Each of at each occurrence is H, methyl, or phenyl. In certain embodiments, G is N. In certain embodiments, the bidentate ligand Lig is 2,2'-bipyridine (bpy) or 4,7-diphenyl-1,10-phenanthroline (BPhen).

[0026] In certain embodiments, some complex compounds of formula I or formula IV are obtained by reacting an intermediate compound of formula II or formula V with a reactive compound of formula III, where M=Ru 2+ , A=Cl - , z=2, y=1, n=2, Lig=2,2'-bipyridine (bpy) or 4,7-diphenyl-1,10-phenanthroline (BPhen), the solvent mixture is a mixture of ethanol and water, and the molar ratio of compound III to compound II or compound V is 1.0 or slightly greater.

[0027] In certain embodiments of the complex compounds of formula I or formula IV, each of the groups P3, P5, P6, Q3, Q4, Q5, Q6, R3 (the definition of R3 relates only to formula I since formula IV does not include R3), R5, R6 and R8 at each occurrence is H or (C1-C3)-alkyl, the group R7 is H, N[(C1-C3)-alkyl)]2 or NO2, alternatively, the groups R6, R7 and R8 together form a bicyclic ring system of the formula shown in claim 1, wherein V1, V2, V3, V4, V5, V6, V7, V8, V9, V 10 , V 11 and V 12 Each of is H.

[0028] In certain embodiments of the complex compound of Formula I or Formula IV, the group R4 is H, (C1-C3)-alkyl, CF3, CH(CH3)OH, CH(CH3)O(C=O)-[CH2]3-Ph, CH(CH3)O(C=O)-[CH2]2-(C=O)-CH2-NH2, or [ka] is a group selected from the group consisting of:

[0029] It is to be noted that in a preferred embodiment of the present invention, formula IV can be obtained from formula I when m≧1 (e.g., 1, 2, 3 or more) and group R3 which is CH2 in formula I forms a six-membered ring with the T′ group which is closest to the coumarin skeleton, provided that T′=CH2. It is further to be noted that in another preferred embodiment of the present invention, formula V can be obtained from formula II when m≧1 (e.g., 1, 2, 3 or more) and group R3 which is CH2 in formula II forms a six-membered ring with the T′ group which is closest to the coumarin skeleton, provided that T′=CH2.

[0030] In certain embodiments, m=0, 1, or 2. In certain embodiments, the complex compound of formula I or formula IV has the following formula Ia, formula Ib, formula Ic, formula Id, formula Ie, formula If, formula Ig, formula Ih, formula Im, formula In, formula Io, formula Ip, formula Iq, formula IVa, or formula IVb: [ka] I have one TIFF2026508498000012.tif253170.

[0031] In certain embodiments, the intermediate compound of formula II (IIa, IIb, IIc, IId, IIe, IIf, IIm, IIn, IIo, IIp, IIq, IIr) or formula V (Va or Vb) has one of the following formulas: IIa, IIb, IIc, IId, IIe, IIf, IIm, IIn, IIo, IIp, IIq, IIr, Va or Vb. [ka] TIFF2026508498000014.tif193170

[0032] Scheme 1: Preparation of complex compounds of formula Ia, formula Ib and formula Ic [ka] Complex Compound of Formula Ia. The ligand compound of Formula IIa (31 mg, 0.073 mmol) and [Ru(bpy)2Cl2] (37 mg, 0.076 mmol) were dissolved in 3 mL of a 3:1 (v / v) solution of EtOH / H2O. The reaction mixture was stirred overnight at 80 °C and analyzed by HPLC-MS to confirm the formation of the product. The mixture was evaporated to dryness, and the resulting aqueous solution was lyophilized. The product was isolated by silica column chromatography, starting with hexane and increasing the polarity of the eluent first with DCM (0% → 100%) and then with MeOH (0% → 20%). 41 mg of a maroon-red solid was obtained (yield: 89%) and identified as the complex compound of Formula Ia. TLC: Rf (35% MeOH in DCM) 0.4. HR-ESI MS (ESI): m / z 418.1265, [C 47 H 42 N8ORu] 2+ Calculated value: 418.1257.

[0033] Complex Compound of Formula Ib. The ligand compound of Formula IIb (34 mg, 0.076 mmol) and [Ru(bpy)2Cl2] (38 mg, 0.078 mmol) were dissolved in 3 mL of a 3:1 (v / v) solution of EtOH / H2O. The reaction mixture was stirred overnight at 80 °C and analyzed by HPLC-MS to confirm the formation of the product. The solvent was evaporated to dryness, and the product was isolated by silica column chromatography, starting with hexane and increasing the polarity of the eluent first with DCM (0% → 100%) and then with MeOH (0% → 20%). 46 mg of a dark maroon solid was obtained (yield: 65%), which was identified as the complex compound of Formula Ib. TLC: Rf (1:9 MeOH / DCM) 0.5. HR-ESI MS (ESI): m / z 430.1258, [C 49 H 42 N8ORu] 2+ Calculated value: 430.1257.

[0034] Complex Compound of Formula Ic. The ligand compound of Formula IIc (31.6 mg, 0.063 mmol) and [Ru(bpy)2Cl2] (37.4 mg, 0.077 mmol) were dissolved in 2 mL of a 3:1 (v / v) solution of EtOH / H2O. The reaction was stirred overnight at 80 °C and analyzed by HPLC-MS to confirm the formation of the product. The solvent was evaporated to dryness, and the product was isolated by silica column chromatography, starting with hexane and increasing the polarity of the eluent first with DCM (0% → 100%) and then with MeOH (0% → 16%). 51.4 mg of a dark purple solid was obtained, identified as the complex compound of Formula Ic (yield: 87%). TLC: Rf (30% MeOH in DCM) 0.4. HR-ESI MS (ESI): m / z 457.1102, [C 49 H 39 N8OF3Ru] 2+ Calculated value: 457.1115.

[0035] Scheme 2: Preparation of complex compounds of formula Id, formula Ie and formula If [ka] Complex Compound of Formula Id. The ligand compound of Formula IId (57.7 mg, 0.096 mmol) and [Ru(bpy)Cl] (55.7 mg, 0.115 mmol) were dissolved in 6 mL of a 3:1 (v / v) solution of EtOH / H2O. A few drops (0.6 mL) of DCM were used to completely dissolve the reagents. The reaction mixture was heated to 80 °C and refluxed overnight. The solvent was evaporated to dryness, and the product was purified by flash chromatography (Puriflash system; first 0% → 100% DCM in hexane, then 0% → 44% MeOH in DCM, Silica column - PF-30SIHP-F0012) to give 34 mg of a maroon-red solid (yield: 34%), which was identified as the complex compound of Formula Id. TLC: Rf (30% MeOH in DCM) 0.43. HR-ESI MS(ESI): m / z 506.1689, [C 58 H 54 N8O3Ru] 2+ Calculated value: 506.1681.

[0036] Complex compound of formula Ie. The ligand compound of formula IIe (21.35 mg, 0.047 mmol) and [Ru(bpy)Cl] (27.32 mg, 0.056 mmol) were dissolved in a 3:1 (v / v) mixture of EtOH and HO (2 mL). A few drops (0.3 mL) of DCM were also added to completely dissolve the reagents. The reaction mixture was reacted at 80 °C overnight. After evaporation of the solvent under reduced pressure, the product was isolated by semi-preparative RP-HPLC (30 min gradient from 10% to 100% B; A, 0.05% TFA in HO; B, 0.1% TFA in ACN; flow rate, 3 mL). After lyophilization, 16.4 mg of a red solid (yield: 32%) was obtained. The chloride salt of the compound, identified as the complex compound of formula Ie, was obtained by using Amberlite IRA402 chloride resin. LR-ESI MS(ESI): m / z 433.34, [C 48 H 44 N8O2Ru] 2+ Calculated value: 433.1315.

[0037] The complex compound of formula If was prepared. [RuCl(bpy)] (18.4 mg, 0.038 mmol) and AgNO (6.5 mg, 0.038 mmol) were dissolved in a 1:1 (v / v) mixture of MeOH and Milli-Q water (2.4 mL) and stirred at 90 °C for 4 h under light-protected conditions. A solution of the ligand compound IIf (28 mg, 0.038 mmol) in THF (1.2 mL) was then added, and the reaction mixture was allowed to react at room temperature for 72 h. Finally, KPF (69.9 mg, 0.38 mmol) was added, and the reaction mixture was allowed to react overnight. After evaporation of the solvent under reduced pressure, the product was isolated by semi-preparative RP-HPLC (30 min gradient from 50% to 100% B; A, 0.05% TFA in HO; B, 0.1% TFA in ACN; flow rate, 3 mL). After lyophilization, 18.8 mg of a red solid was obtained (yield: 36%). TLC: f (30% MeOH in DCM) = 0.48. The chloride salt of the compound was identified as the complex compound of formula If and was obtained by using Amberlite IRA402 chloride resin. LR-ESI MS (ESI): m / z 575.81, [C 62 H 61 Cl2N9O3Ru] 2+ Calculated value: 575.6659.

[0038] Scheme 3: Preparation of complex compounds of formula Ig and formula Ih [ka] Complex Compound of Formula Ig. The ligand compound of Formula IIa (20.4 mg, 0.048 mmol) and [Ru(BPhen)2Cl2] (49.5 mg, 0.059 mmol) were dissolved in 3 mL of a 3:1 (v / v) solution of EtOH / H2O. The reaction mixture was stirred overnight at 80 °C and analyzed by HPLC-MS to confirm the formation of the product. The solution mixture was evaporated to dryness and the product was isolated by silica column chromatography, starting with 100% DCM and increasing the polarity of the eluent with MeOH (0% to 6%). 37.3 mg of a red solid was obtained (yield: 63%), which was identified as the complex compound of Formula Ig. TLC: Rf (10% MeOH in DCM) 0.4. HR-ESI MS (positive mode): m / z 594.1892, [C75 H 58 N8ORu] 2+ Calculated value: 594.1888.

[0039] Complex Compound of Formula Ih. The ligand compound of Formula IIc (11.0 mg, 0.022 mmol) and [Ru(BPhen)2Cl2] (41.9 mg, 0.050 mmol) were dissolved in 8 mL of a 3:1 (v / v) solution of EtOH / H2O. The reaction mixture was stirred overnight at 80 °C and analyzed by HPLC-MS to confirm the formation of the product. The solution mixture was evaporated to dryness and the product was isolated by silica column chromatography, starting with hexane and increasing the polarity of the eluent first with DCM (0% → 100%) and then with MeOH (0% → 26%). 23.7 mg of a maroon-red solid was obtained (yield: 81%) and identified as the complex compound of Formula Ih. TLC: Rf (10% MeOH in DCM) 0.3. HR-ESI MS (positive mode): m / z 633.1740, [C 77 H 55 F3N8ORu] 2+ Calculated value: 633.1741.

[0040] Scheme 4: Preparation of complex compounds of formula Ii and formula Ij [ka] Complex compound of formula II. To a solution of [Os(bpy)2Cl2] (10 mg, 0.017 mmol) in ethylene glycol, the ligand compound of formula IIa (8 mg, 0.019 mmol) was added. The mixture was heated at 85 °C under N2 for 18 h. The solution was cooled to room temperature. After adding a saturated aqueous solution of ammonium hexafluorophosphate, the resulting dark purple precipitate was isolated by filtration, washed with water, diethyl ether, and dried under vacuum. The crude product was then purified by column chromatography (silica gel, 0% → 15% MeOH in CHCl2). After converting the PF6 salt to the chloride salt using Amberlite IRA 410 chloride resin, 8 mg of a dark purple solid was obtained (42% yield). TLC: Rf (10% MeOH in CHCl2) 0.45. HR-ESI MS (positive mode): m / z 463.1542, [C47 H 42 N8OOs] 2+ Calculated value: 463.1542.

[0041] Complex compound of formula Ij. To a solution of [Os(bpy)2Cl2] (14 mg, 0.024 mmol) in ethylene glycol, the ligand compound of formula IIc (12 mg, 0.028 mmol) was added. The mixture was heated at 85 °C under N2 for 18 h. The solution was cooled to room temperature. After adding a saturated aqueous solution of ammonium hexafluorophosphate, the resulting dark purple precipitate was isolated by filtration, washed with water, diethyl ether, and dried under vacuum. The crude product was then purified by column chromatography (silica gel, 0% → 15% MeOH in CHCl2). After converting the PF6 salt to the chloride salt using Amberlite IRA 410 chloride resin, 7 mg of a dark purple solid was obtained (28% yield). Characterization: TLC: Rf (10% MeOH in CHCl2) 0.40. HR-ESI MS (positive mode): m / z 502.1402, [C 51 H 45 F3N8OOs] 2+ Calculated value: 502.1401.

[0042] Scheme 5: Preparation of complex compounds of formula Ik [ka] Complex compound of formula Ik. To a degassed ethylene glycol solution of [Os(BPhen)2Cl2] (30 mg, 0.032 mmol) was added the ligand compound of formula IIa (14.8 mg, 0.035 mmol). The mixture was heated at 110 °C under N2 for 18 h. The solution was cooled to room temperature. After adding a saturated aqueous solution of ammonium hexafluorophosphate, the resulting dark purple precipitate was isolated by filtration, washed with water, diethyl ether, and dried under vacuum. The crude product was first purified by column chromatography (silica gel, 0% → 15% MeOH in CHCl2) and then by preparative HPLC (30 min gradient from 0% → 100% B, flow rate: 10 mL / min, R tFurther purification was achieved by HPLC (HPLC = 15.4 min). After conversion of the PF6 salt to the chloride salt using Amberlite IRA 410 chloride resin, 9 mg of a dark purple solid was obtained (22% yield). TLC: Rf (10% MeOH in CH2Cl2) 0.51. HR-ESI MS (positive mode): m / z 639.2181, [C 75 H 58 N8OOs] 2+ Calculated value: 639.2169.

[0043] Scheme 6: Preparation of complex compounds of formula Im [ka] Complex Compound of Formula Im. The ligand compound of Formula IIm (31.6 mg, 0.060 mmol) and [Ru(bpy)2Cl2] (37.4 mg, 0.077 mmol) were dissolved in a 3:1 (v / v) solution of EtOH / H2O (2 mL), and the reaction mixture was stirred at 90 °C overnight. The solvent was evaporated to dryness, and the product was purified by silica column chromatography, starting with hexane and increasing the polarity of the eluent first with DCM (0% → 100%) and then with MeOH (0% → 11%). 10 mg of a blue solid was obtained (yield: 17%), which was identified as the complex compound of Formula Im. TLC: Rf (30% MeOH in DCM) 0.3. HR-ESI MS (ESI): m / z 470.1193, [C 51 H 41 F3N8ORu] 2+ Calculated value: 470.1199.

[0044] Scheme 7: Preparation of ligand compounds of formula IIa, IIb and IIc [ka] 4-Methyl-4'-((trimethylsilyl)methyl)-2,2'-bipyridine of Formula 1. A solution of 4,4'-dimethyl-2,2'-bipyridine (2 g, 10.86 mmol) in anhydrous THF (80 mL) was added dropwise via cannula under an Ar atmosphere to a cooled (-78 °C) solution of LDA in THF (12 mL, 11.95 mmol). The resulting maroon mixture was stirred at -78 °C for 1 h. Trimethylsilyl chloride (2 mL, 11.95 mmol) was then added to the crude product, which turned blue, and exactly 10 s later, 10 mL of absolute ethanol was carefully added, turning the solution yellow. The crude product was transferred to a separatory funnel containing a saturated solution of NaHCO (200 mL) and extracted with DCM (3 × 150 mL). The combined organic phase was washed with brine (150 mL), dried over anhydrous Na2SO4, filtered and evaporated to dryness to give 2.12 g of the title compound as a yellow solid (yield: 76%), which was used without further purification.

[0045] 4-(Chloromethyl)-4'-methyl-2,2'-bipyridine of Formula 2. 4-Methyl-4'-((trimethylsilyl)methyl)-2,2'-bipyridine (2.12 g, 7.80 mmol), hexachloroethane (7.39 g, 31.2 mmol), and cesium fluoride (4.76 g, 31.2 mmol) were dissolved in anhydrous ACN (120 mL) under an Ar atmosphere, and the resulting solution was stirred at 60 °C for 3.5 h. The reaction mixture was partitioned between 50 mL of HO and 50 mL of AcOEt and transferred to a separatory funnel. The aqueous phase was extracted with ethyl acetate (3 × 50 mL), and the combined organic phases were washed with brine (50 mL), dried over anhydrous NaSO, filtered, and evaporated to dryness. The product was purified by column chromatography (silica gel, 0%→10% MeOH in DCM) to give 1.01 g of the title compound as a yellow solid (yield: 57%). TLC: Rf (1:9 MeOH / DCM) 0.6. LR-ESI MS (ESI): m / z 218.8, [C 12 H 11 N2Cl+H] + Calculated value: 218.1.

[0046] 2-(4'-Methyl-[2,2'-bipyridin]-4-yl)acetonitrile of Formula 3. 4-(Chloromethyl)-4'-methyl-2,2'-bipyridine (1 g, 4.57 mmol), 18-crown-6 (26.8 mg, 0.09 mmol), and KCN (3.39 g, 36.5 mmol) were dissolved in 100 mL of ACN and stirred overnight at room temperature. HPLC-MS analysis of the crude product revealed little product formation. 214 mg of 18-crown-6 (0.81 mmol) was then added, and the solution was stirred overnight at 50 °C. After HPLC-MS showed the reaction was complete, the reaction mixture was evaporated to dryness, redissolved in deionized water (100 mL), and extracted with DCM (3 × 100 mL). The combined organic phases were dried over anhydrous Na2SO4, filtered, and evaporated to dryness. After purification by column chromatography (silica gel, 50%→100% AcOEt in hexane), 615 mg of the title compound was obtained as a white solid (yield: 64%). TLC: Rf (AcOEt 100%) 0.7. HR-ESI MS (ESI): m / z 210.1026, [C 13 H 11 N3+H] + Calculated value: 210.1025.

[0047] Thiocoumarin of Formula 4. 7-Diethylamino-4-methylcoumarin (1.72 g, 7.42 mmol) and Lawesson's reagent (1.65 g, 4.08 mmol) were dissolved in toluene (40 mL) and heated at 100° C. for 12 h. After evaporation under reduced pressure, the dark residue was purified by column chromatography (silica gel, DCM) to give thiocoumarin of Formula 4 as an orange solid (1.70 g, 92%). TLC: Rf (DCM) 0.50. HRMS (ESI-TOF) m / z [M+H] + , C 14 H 18 NOS calculated value: 248.1109, measured value: 248.1099.

[0048] Thiocoumarin of Formula 5. 2,3,6,7-Tetrahydro-9-methyl-1H,5H-quinolizino[9,1-gh]coumarin (2 g, 7.8 mmol) and Lawesson's reagent (1.89 g, 4.7 mmol) were dissolved in 100 mL of toluene and stirred at 100° C. overnight. The dark green crude material was evaporated to dryness and the product was isolated by column chromatography (silica gel, 0% to 70% DCM in hexane). 1.44 g of thiocoumarin of Formula 5 was obtained as an orange solid (yield: 68%). TLC: Rf (100% DCM) 0.8. HR-ESI MS (ESI): m / z 272.1104, [C 16 H 17 NOS+H] + Calculated value: 272.1102.

[0049] Thiocoumarin of Formula 6. 2,3,6,7-Tetrahydro-9-trifluoromethyl-1H,5H-quinolizino[9,1-gh]coumarin (1 g, 3.23 mmol) and Lawesson's reagent (791 mg, 1.94 mmol) were dissolved in toluene (50 mL). The yellow solution was stirred at 100°C overnight. The dark red solution was evaporated to dryness and purified by column chromatography (silica gel, 0% to 40% DCM in hexanes) to isolate thiocoumarin of Formula 6. 1.14 g of a golden maroon solid was obtained (yield: 95%). Golden maroon solid. TLC: Rf (100% DCM) 0.9. HR-ESI MS (ESI): m / z 326.0821, [C 16 H 14 F3NOS+H] + Calculated value: 326.0816.

[0050] Ligand compound of Formula IIa. To a solution of sodium hydride (568 mg, 14.15 mmol, 60% dispersion in mineral oil) and compound 3 (712 mg, 3.40 mmol) in anhydrous acetonitrile (300 mL) was added 7-(N,N-diethylamino-4-methyl-2-thiocoumarin) (703 mg, 5.66 mmol). The orange solution was stirred for 3 h. Silver nitrate (962 mg, 4.98 mmol) was then added, and the reaction mixture was stirred at room temperature for 2 h. The dark maroon solution was analyzed by HPLC-MS to confirm the formation of the desired product in the crude, which was evaporated under reduced pressure. The product was isolated by silica column chromatography, starting with hexane and increasing the polarity of the eluent first with DCM (0% → 100%) and then with MeOH (0% → 10%). 451 mg of an orange / red solid was obtained (yield: 37%) and identified as the ligand compound of Formula IIa. TLC: Rf (1:9 MeOH / DCM)0.5. HR-ESI MS(ESI): m / z 423.2181, [C 27 H 26 NO+H] + Calculated value: 423.2179.

[0051] Ligand compound of formula IIb. To a solution of NaH (38 mg, 1.58 mmol, 60% dispersion in mineral oil) and compound 3 (46 mg, 0.22 mmol) in anhydrous ACN (12 mL) was added thiocoumarin of formula 5 (50 mg, 0.18 mmol). The maroon solution was stirred at 50°C for 3 h. Silver nitrate (71 mg, 0.41 mmol) was then added, and the crude was stirred at room temperature for 2 h. The product was isolated by silica column chromatography, starting with hexane and increasing the eluent polarity first with DCM (0% → 100%) and then MeOH (0% → 10%). 20 mg of an orange / brown solid was obtained (yield: 20%) and identified as the ligand compound of formula IIb. TLC: Rf (5% MeOH / DCM) 0.5. HR-ESI MS (ESI): m / z 447.2186, [C 29 H 26 NO+H] + Calculated value: 447.2179.

[0052] Ligand compound of formula IIc. To a solution of NaH (18 mg, 0.77 mmol of a 60% dispersion in mineral oil) and compound 3 (41 mg, 0.18 mmol) in anhydrous ACN (35 mL) was added thiocoumarin of formula 6 (50 mg, 0.15 mmol). The maroon solution was stirred for 3 h. Silver nitrate (56 mg, 0.32 mmol) was then added, and the crude was stirred at room temperature for 2 h. The product was isolated by silica column chromatography, starting with hexane and increasing the eluent polarity first with DCM (0% → 100%) and then MeOH (0% → 20%). 58 mg of a bright red solid was obtained (yield: 75%) and identified as the ligand compound of formula IIc. Bright red solid. TLC: Rf (5% MeOH / DCM) 0.5. HR-ESI MS (ESI): m / z 501.1889, [C 29 H 23 F3N4O+H] + Calculated value: 501.1887.

[0053] Scheme 8: Preparation of Ligand Compounds of Formula IId, IIe, and IIf [ka] Compound of Formula 11. A mixture of coumarin of Formula 10 (1.60 g, 6.12 mmol), phenylbutyric acid (1.50 g, 9.15 mmol), EDC (1.75 g, 9.15 mmol), and 4-dimethylaminopyridine (DMAP) (1.12 g, 9.15 mmol) was cooled to 0 °C under an argon atmosphere and then dissolved in DCM (100 mL). The mixture was stirred at 0 °C for 15 minutes and then at room temperature for 17 hours. The solution was then washed with saturated NH4Cl (2 × 100 mL), 5% aqueous NaHCO3 (1 × 100 mL, 2 × 50 mL), and deionized water (100 mL). The organic layer was dried over anhydrous MgSO4, filtered, and evaporated under reduced pressure. The product was isolated by silica column chromatography (silica gel, 50%→100% DCM in hexanes, 1%→3% MeOH in DCM) to give 2 g of the title compound as a yellow / orange solid (yield: 80%). TLC: Rf (DCM) 0.6. HR-ESI MS (ESI): m / z 408.2169, [C 25 H 29NO4 + H] + Calculated value: 408.2175.

[0054] Compound of Formula 12. Lawesson's reagent (1.54 g, 3.81 mmol) was added to a solution of compound of Formula 11 (1.94 g, 4.75 mmol) in toluene (60 mL) under an Ar atmosphere. The mixture was stirred overnight at 105 °C in the dark. A color change from dark brown to pale yellow was observed. After removing the solvent under reduced pressure, the product was isolated by column chromatography (silica gel, 100% → 30% hexane in DCM) to give 1.67 g of the title compound as an orange solid (yield: 83%). TLC:R f (DCM)=0.78. HR-ESI MS(ESI): m / z 424.1936, [C 25 H 29 NO3S+H] + Calculated value: 424.1946.

[0055] Ligand compound of Formula IId. First, sodium hydride (68.3 mg, 1.71 mmol, 60% dispersion in mineral oil) and 2-(4'-methyl-[2,2'-bipyridin]-4-yl)acetonitrile (Formula 3) (357 mg, 1.71 mmol) were dissolved in anhydrous ACN (30 mL) and stirred at 35 °C for 15 min under an argon atmosphere. Next, a solution of coumarin of Formula 12 (362 mg, 0.85 mmol) in anhydrous ACN (15 mL) was added to the previous flask, and the resulting reaction mixture was stirred at 35 °C for an additional 4 h under an argon atmosphere in the dark. After 4 h, AgNO (363 mg, 2.14 mmol, 2.5 equiv.) was added and stirred for an additional 2 h. A color change from brown to dark red / maroon was observed. After evaporation of the solvent under reduced pressure, the product was isolated by silica column chromatography, starting with a 1:1 mixture of DCM and hexane, increasing the polarity of the eluent by 10-fold increments with DCM up to 100%, and then increasing the polarity of the eluent with MeOH in DCM (0% to 1%). 155 mg of an orange / red solid was obtained (yield: 30%), which was identified as the ligand compound of formula IId. TLC:R f (10%MeOH in DCM)=0.4. HR-ESI MS(ESI): m / z 599.3014, [C 38 H 38N4O3+H] + Calculated value: 599.3022.

[0056] Ligand compound of Formula IIe. To a solution of the ligand compound of Formula IId (41.7 mg, 0.070 mmol) in 2:1 (v / v) ACN / HO (7 mL), 10% NaOH (6.65 mL, 0.070 mmol) was added, and the reaction mixture was stirred overnight at room temperature in the dark. The reaction mixture changed color from orange / yellow to bright orange and contained solid particles. After evaporation to dryness under reduced pressure, the crude material was purified by flash chromatography (DCM in hexane (50% → 100%), then MeOH in DCM (0% → 10%), Puriflash system, Silica column PF-DLE-F0012) to give 26.9 mg of an orange / red solid (85% yield), which was identified as the ligand compound of Formula IIe. TLC: Rf (10% MeOH in DCM) 0.57. HR-ESI MS(ESI): m / z 452.56, [C 29 H 28 N4O2+H] + Calculated value: 453.2291.

[0057] Ligand compound of Formula IIf. Chlorambucil (46.54 mg, 0.153 mmol), 4-dimethylaminopyridine (DMAP) (18.69 mg, 0.153 mmol), and 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl) (29.33 mg, 0.153 mmol) were added to a round-bottom flask containing compound of Formula 14 (46.10 mg, 0.102 mmol) under an argon atmosphere and placed in an ice bath. The reagents were dissolved in DCM (13 mL), and the reaction mixture was stirred for 15 minutes. The flask was then removed from the ice bath and allowed to react overnight at room temperature in the dark. Upon completion of the reaction (as confirmed by TLC and HPLC), DCM was evaporated under reduced pressure, followed by purification by column chromatography (silica gel, 0% to 100% DCM in hexane, then 0% to 1% MeOH in DCM) to give 49.9 mg of an orange / red solid (yield: 66%), identified as the ligand compound of formula IIf. TLC: Rf (10% MeOH in DCM) 0.75. HR-ESI MS (ESI): m / z 739.85, [C 42 H 45 Cl2N5O3+H] + Calculated value: 738.2978.

[0058] Scheme 9: Preparation of ligand compounds of formula IIm [ka] Nitrile coumarin of Formula 16. To a cooled solution of CHCN (0.7 mL, 13.4 mmol) in anhydrous THF (40 mL) at −78 °C, n-BuLi (5.2 mL, 2.5 M in hexane, 13 mmol) was added under Ar atmosphere. The resulting clear, colorless solution was stirred at −78 °C for 15 min, after which the appearance of a white suspension was observed. A solution of 2,3,6,7-tetrahydro-9-trifluoromethyl-1H,5H-quinolizino[9,1-gh]coumarin (1.023 g, 3.31 mmol) in anhydrous THF (35 mL) was then slowly added, and the reaction mixture was stirred at −78 °C for 30 min under Ar. The resulting orange mixture was quenched at −78 °C by the addition of saturated aqueous NHCl (35 mL), resulting in the precipitation of a white solid. The mixture was warmed to room temperature and extracted with AcOEt (3 × 50 mL). The combined organic phases were evaporated to dryness. To the resulting crude oil, 0.5 M aqueous HCl (200 mL) was added and the mixture was stirred vigorously for 3 h. The color of the solution changed from orange to brown. The solution was extracted with AcOEt (3 × 80 mL), dried over anhydrous MgSO4, filtered, and evaporated to dryness under reduced pressure. The crude mixture was purified by flash chromatography (silica gel, 0% → 15% AcOEt in hexane). 760 mg of the title compound was obtained as a light orange solid (yield: 70%). Orange solid. TLC: Rf (25% AcOEt in hexane) 0.38. HR-MS (ESI): m / z 332.1209, [C 18 H 15 F3N2O+H] + Calculated value: 333.1215.

[0059] Coumarin aldehyde of formula 17. Nitrile coumarin of formula 16 (300 mg, 0.90 mmol) was dissolved in anhydrous toluene (35 mL) under an Ar atmosphere, and the mixture was stirred for 10 minutes (orange solution). Then, a 1 M solution of DIBALH in toluene (1.35 mL, 1.35 mmol) was added, and the reaction mixture was stirred at room temperature under an Ar atmosphere for 30 minutes (the color changed to maroon). The crude was cooled in an ice bath, and acetone (4 mL) was added to destroy the excess reagent (the color changed from maroon to dark red). After the addition of saturated sodium potassium tartrate (25 mL), the crude was extracted with AcOEt (3 × 25 mL), and the organic phase was washed with water (25 mL) and brine (25 mL), dried over anhydrous MgSO4, filtered, and evaporated to dryness under reduced pressure. The compound was purified by silica column chromatography using hexane, increasing polarity first with DCM (0% to 100%) and then with MeOH (0% to 0.5%). 150 mg of the title compound was obtained as a red solid (yield: 50%). TLC: Rf (5% MeOH in DCM) 0.74. HR-MS (ESI): m / z 336.1206, [C 18 H 16 F3NO2+H] + Calculated value: 336.1211.

[0060] Ligand compound of formula IIm. To a solution of coumarin aldehyde of formula 17 (50 mg, 0.15 mmol) in absolute ethanol (7 mL) were added piperidine (70 mg, 0.83 mmol) and 2-(4'-methyl-[2,2'-bipyridin]-4-yl)acetonitrile (46 mg, 0.22 mmol). The reaction mixture (dark maroon color) was stirred at 80 °C overnight. After evaporation to dryness under reduced pressure, the compound was purified by silica column chromatography using hexane and increasing polarity first with DCM (0% → 100%) and then with MeOH (0% → 2%). 66 mg of a violet solid was obtained (yield: 83%), identified as the ligand compound of formula IIm. TLC: Rf (5% MeOH in DCM) 0.4. HR-ESI MS (ESI): m / z 527.2077, [C 31 H 25 F3N4O+H] + Calculated value: 527.2053.

[0061] Scheme 10: Preparation of ligand compounds of formula IIo [ka] Nitrile coumarin of formula 18. To a solution of coumarin aldehyde of formula 17 (207 mg, 0.62 mmol) in toluene (15 mL) was added (triphenylphosphoranylidene)acetonitrile (850 mg, 2.82 mmol). After degassing the solution with Ar, the vial was sealed, and the mixture was stirred at 60 °C for 7 days. Upon complete consumption of the starting aldehyde, deionized water (HO) was added to the crude product, followed by extraction with AcOEt (3 × 30 mL). The combined organic phases were dried over anhydrous MgSO, filtered, and evaporated to dryness under reduced pressure. The crude product was purified by silica column chromatography using hexane and increasing the polarity with AcOEt (0% → 12%). 184 mg of the title compound was obtained as a red solid (yield: 84%). TLC: Rf (50% AcOEt in hexane) 0.90. HR-MS(ESI): m / z 359.1366, [C 20 H 17 F3N2O+H] + Calculated value: 359.1357.

[0062] Coumarin aldehyde of Formula 19. Nitrile coumarin of Formula 18 (103 mg, 0.29 mmol) was dissolved in anhydrous THF (30 mL) under an Ar atmosphere, and the mixture was cooled in an ice bath. A 1 M solution of DIBALH in toluene (3.0 mL, 2.95 mmol) was then added, and the reaction mixture was stirred at room temperature under an Ar atmosphere for 30 min. The crude was cooled in an ice bath, and acetone (10 mL) was added to destroy excess reagent. After adding saturated potassium sodium tartrate (25 mL) and stirring for 30 min, the crude was extracted with DCM (3 × 50 mL). The organic phase was washed with brine (50 mL), dried over anhydrous MgSO4, filtered, and evaporated to dryness under reduced pressure. The compound was purified by silica column chromatography using hexane, first increasing polarity with DCM (0% → 100%), then MeOH (0% → 0.5%). 78 mg of the title compound was obtained as a yellow solid (yield: 69%). TLC:Rf (5% MeOH in DCM) 0.52. HR-MS(ESI): m / z 362.1362, [C 20 H 17 F3N2O+H] + Calculated value: 362.1359.

[0063] Ligand compound of formula IIo. To a solution of coumarin aldehyde of formula 19 (32 mg, 0.088 mmol) in absolute ethanol (10 mL) were added piperidine (45 mg, 0.531 mmol) and 2-(4'-methyl-[2,2'-bipyridin]-4-yl)acetonitrile (23 mg, 0.11 mmol). The reaction mixture was stirred at 80°C overnight. After evaporation to dryness under reduced pressure, the compound was purified by silica column chromatography using hexane, first with DCM (0% to 100%) and then with MeOH (0% to 1%), increasing polarity. 44 mg of a violet solid was obtained (yield: 89%), identified as the ligand compound of formula IIo. TLC: Rf (5% MeOH in DCM) 0.59. LR-MS (ESI): m / z 553.25, [C 33 H 27 F3N4O+H] + Calculated value: 553.2210.

[0064] Scheme 11: Preparation of ligand compounds of formula IIn [ka] Nitrile coumarin of Formula 20. To a cooled solution (-78 °C) of CHCN (2.42 mL, 34.6 mmol) in anhydrous THF (40 mL), n-BuLi (13.8 mL, 2.5 M in hexane, 34.6 mmol) was added under Ar. The resulting clear, colorless solution was stirred at -78 °C for 15 min, after which the appearance of a white suspension was observed. A solution of 7-(diethylamino)-4-methyl-2H-chromen-2-one coumarin (2.008 g, 8.65 mmol) in anhydrous THF (30 mL) was then slowly added, and the reaction mixture was stirred at -78 °C for 30 min under Ar. The resulting white mixture was quenched at -78 °C by the addition of saturated aqueous NH4Cl (35 mL), resulting in the precipitation of a white solid. The mixture was warmed to room temperature and extracted with AcOEt (3 × 50 mL). The combined organic phase was evaporated to dryness. To the resulting orange crude oil was added 0.5 M aqueous HCl (200 mL) and the mixture was stirred vigorously for 16 h. The solution turned brown and a yellow solid precipitated. 50 mL of DCM was added to dissolve the precipitate, and the mixture was extracted with DCM (3 × 100 mL), dried over anhydrous MgSO4, filtered, and evaporated to dryness under reduced pressure to give a brown solid. The crude mixture was purified by silica column chromatography (0% → 22% AcOEt in hexane). 2.01 g of the title compound was obtained as a light orange solid (yield: 92%). Orange solid. TLC: Rf (50% AcOEt in hexane) 0.61. LR-ESI MS (ESI): m / z 255.2, [C 16 H 18 NO+H] + Calculated value: 255.1492.

[0065] Coumarin aldehyde of Formula 21. Nitrile coumarin of Formula 20 (500 mg, 1.97 mmol) was dissolved in anhydrous toluene (25 mL) under an Ar atmosphere, and the mixture was stirred for 10 min (orange solution). A 1 M solution of DIBALH in toluene (3.0 mL, 2.95 mmol) was then added, and the reaction mixture was stirred at room temperature under an Ar atmosphere for 30 min. The crude was cooled in an ice bath, and acetone (10 mL) was added to destroy excess reagent. After the addition of saturated sodium potassium tartrate (40 mL), the crude was extracted with DCM (3 × 50 mL). The organic phase was washed with brine (50 mL), dried over anhydrous MgSO4, filtered, and evaporated to dryness under reduced pressure. The compound was purified by silica column chromatography using hexane, first with DCM (0% → 100%) and then increasing the polarity with MeOH (0% → 0.5%). 78 mg of the title compound was obtained as a yellow solid (yield: 15%). TLC:Rf (5% MeOH in DCM) 0.52. LR-ESI MS(ESI): m / z 258.2, [C 16 H 19 NO2 + H] + Calculated value: 258.1489.

[0066] Ligand compound of formula IIn. To a solution of coumarin aldehyde of formula 21 (47 mg, 0.18 mmol) in absolute ethanol (10 mL) were added piperidine (84 mg, 1.0 mmol) and 2-(4'-methyl-[2,2'-bipyridin]-4-yl)acetonitrile (44 mg, 0.20 mmol). The reaction mixture was stirred at 80°C overnight. After evaporation to dryness under reduced pressure, the compound was purified by silica column chromatography using hexane and increasing polarity first with DCM (0% to 100%) and then with MeOH (0% to 2%). 79 mg of a violet solid was obtained (yield: 96%), identified as the ligand compound of formula IIn. TLC: Rf (5% MeOH in DCM) 0.38. HR-ESI MS (ESI): m / z 449.2336, [C 29 H 28 NO+H] + Calculated value: 449.2336.

[0067] Scheme 12: Preparation of ligand compounds of formula IIp [ka] The ligand compound of Formula IIp was prepared. First, sodium hydride (66 mg, 1.64 mmol, 60% dispersion in mineral oil) and 2-(4'-methyl-[2,2'-bipyridin]-4-yl)acetonitrile (106 mg, 0.506 mmol) were dissolved in anhydrous THF (15 mL) and stirred at 40 °C under an Ar atmosphere for 15 min. Next, a solution of 4-methyl-2H-chromene-2-thione (85 mg, 0.482 mmol) in anhydrous THF (15 mL) was added to the flask, and the resulting reaction mixture was stirred at 40 °C for an additional 4 h in the dark under an Ar atmosphere. After 4 h, AgNO (163 mg, 0.964 mmol) was added and stirred at 35 °C for an additional 2 h. After evaporation of the solvent under reduced pressure, the compound was purified by silica column chromatography using hexane and increasing polarity first with DCM (0% → 100%) and then with MeOH (0% → 1%). 92 mg of an orange solid was obtained (yield: 54%) and was identified as the ligand compound of formula IIp. TLC: Rf (2.5% MeOH in DCM) 0.25. HR-MS (ESI): m / z 352.1451, [C 23 H 17 NO+H] + Calculated value: 352.1444.

[0068] Scheme 13: Preparation of ligand compounds of formula IIq and IIr and complex compounds of formula Iq [ka] Ligand compound of formula IIq. 4-Dimethylaminopyridine (DMAP) (20 mg, 0.164 mmol) and 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl) (38 mg, 0.198 mmol) were added sequentially to an ice-cooled solution of N-Boc-5-aminolevulinic acid (38 mg, 0.164 mmol) in DCM (10 mL) under an argon atmosphere. After stirring the mixture at 0 °C for 15 min, a solution of compound of formula IIe (61 mg, 0.135 mmol) in DCM (5 mL) was added dropwise at the same temperature. The ice bath was then removed, and the resulting mixture was further stirred at room temperature for 16 h. After completion of the reaction (TLC, HPLC), the volatiles were concentrated under reduced pressure and the resulting crude was purified by flash column chromatography (silica gel, MeOH 0% to 5% in DCM) to give 31 mg of an orange / red solid (yield: 34%), which was identified as the ligand compound of formula IIq. LR-MS (ESI): m / z [M+H] + , C 38 H 43 Calculated value of N5O6: 666.33, Measured value: 666.57.

[0069] Ligand compound of formula IIr. To an ice-cooled solution of the ligand compound of formula IIq (15 mg, 0.022 mmol) in anhydrous THF (2 mL) was added 4N HCl in dioxane (675 μL, 2.70 mmol), and the resulting mixture was warmed to room temperature and further stirred for 5 h. After completion of the reaction (HPLC-MS), the reaction mixture was neutralized with 10% (w / v) aqueous NaHCO3 (10 mL), diluted with water (20 mL), extracted with DCM (3 × 20 mL), and the combined organic extracts were dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure to give the crude ligand compound of formula IIr (12 mg, yield: 94%) as an orange / red solid, which was used in the subsequent step without any further purification. LR-MS (ESI): m / z [M+H] + , C 33 H 35 Calculated N5O4 value: 566.28, Measured value: 566.45.

[0070] Complex Compound of Formula Iq. Method A: The ligand compound of Formula IIq (16 mg, 0.024 mmol) and [Ru(bpy)Cl] (10 mg, 0.021 mmol) were dissolved in 4 mL of a 3:1 (v / v) solution of EtOH / HO under an Ar atmosphere. The reaction mixture was stirred at 80 °C overnight and analyzed by HPLC-MS to confirm the formation of the product. The reaction mixture was evaporated to dryness, and the residue was dissolved in anhydrous THF (2 mL). It was treated with 4 N HCl in dioxane (600 μL, 2.37 mmol) at 0 °C, warmed to room temperature, and further stirred for 5 h. After completion of the reaction (HPLC-MS), the reaction mixture was neutralized with 10% (w / v) aqueous NaHCO (5 mL), diluted with water (20 mL), and extracted with DCM (3 × 20 mL). The combined organic extracts were dried over anhydrous MgSO, filtered, and concentrated under reduced pressure to give the crude product. Method B: The ligand compound of formula IIr (11 mg, 0.019 mmol) and [Ru(bpy)Cl] (9 mg, 0.019 mmol) were dissolved in 4 mL of a 3:1 (v / v) solution of EtOH / HO under an Ar atmosphere. The reaction mixture was stirred overnight at 80 °C and analyzed by HPLC-MS to confirm the formation of the product. The reaction mixture was evaporated to dryness to give the crude product. In both methods, the crude product was purified by preparative HPLC (C18 column; mobile phase, A: 0.1% HCOOH in HO, B: 0.1% HCOOH in ACN; gradient: 10% to 80% B in 30 min) to give the desired complex compound of formula Iq as a maroon solid (Method A: 8 mg, yield: 39%; Method B: 10 mg, yield: 55%). Analytical HPLC (C18 column; mobile phase, A: 0.1% HCOOH in H2O, B: 0.1% HCOOH in ACN; gradient: 5% in 5 → 100% B): Rt = 2.14 min. LR-MS(ESI):m / z [M] 2+ , C 53 H 51 N9O4Ru 2+ Calculated value: 489.65, measured value: 489.60.

[0071] Scheme 14: Preparation of ligand compounds of formula Va and formula Vb [ka] Ligand compound of formula Va. To a solution of coumarin aldehyde of formula 22 (92 mg, 0.32 mmol) in absolute ethanol (10 mL) were added piperidine (166 mg, 2.0 mmol) and 2-(4'-methyl-[2,2'-bipyridin]-4-yl)acetonitrile (68 mg, 0.32 mmol). The reaction mixture was stirred at 80°C overnight. After evaporation to dryness under reduced pressure, the compound was purified by silica column chromatography using hexane and increasing the polarity first with DCM (0% to 100%) and then with MeOH (0% to 1%). 73 mg of a violet solid was obtained (yield: 47%), identified as the ligand compound of formula Va. TLC: Rf (5% MeOH in DCM) 0.59. HR-MS (ESI): m / z 475.2495, [C 31 H 30 NO+H] + Calculated value: 475.2492.

[0072] Ligand compound of formula Vb. To a solution of coumarin aldehyde of formula 23 (40 mg, 0.155 mmol) in absolute ethanol (8 mL) were added piperidine (79 mg, 0.932 mmol) and 2-(4'-methyl-[2,2'-bipyridin]-4-yl)acetonitrile (33 mg, 0.155 mmol). The reaction mixture was stirred at 80°C overnight. After evaporation to dryness under reduced pressure, the compound was purified by silica column chromatography using hexane, first with DCM (0% → 100%) and then with MeOH (0% → 2%), increasing polarity. 32 mg of a brown solid was obtained (yield: 46%), identified as the ligand compound of formula Vb. TLC: Rf (5% MeOH in DCM) 0.30. LR-MS (ESI): m / z 449.07, [C 27 H 20 N4O3+H] + Calculated value: 449.1608.

[0073] General Procedure for the Synthesis of Complex Compounds of Formula In, Formula Io, Formula Ip, Formula IVa and Formula IVb The corresponding ligand compound of Formula IIn, Formula IIo, Formula IIp, Formula Va, or Formula Vb and [Ru(bpy)2Cl2] (1.1 equivalents) were dissolved in a 1:1 (v / v) mixture of EtOH / HO, and the reaction mixture was stirred at 80-90 °C for 24-48 h under an argon atmosphere. The reaction was monitored by HPLC-MS. After complete conversion, the solvent was evaporated to dryness, and the product was purified by silica column chromatography using DCM and increasing the eluent polarity with MeOH. The compound was obtained as a racemic mixture of chloride salts and characterized by HR-ESI MS.

[0074] In:HR-ESI MS(ESI):m / z 431.1336, [C 49 H 44 N8ORu] 2+ Calculated for: 431.1336. Analytical HPLC (5% → 100% B in 5 min, with formic acid): Rt = 2.46 min.

[0075] Io:HR-ESI MS(ESI):m / z 483.1282, [C 53 H 43 F3N8ORu] 2+ Calculated for: 483.1272. Analytical HPLC (5% → 100% B in 5 min, with formic acid): Rt = 2.88 min.

[0076] Ip:HR-ESI MS(ESI):m / z 382.5898, [C 43 H 33 N7ORu] 2+ Calculated for: 382.5890. Analytical HPLC (5% → 100% B in 5 min, with formic acid): Rt = 2.25 min.

[0077] IVa:HR-ESI MS(ESI):m / z 444.1420, [C 51 H 44 N8ORu] 2+ Calculated for: 444.1414. Analytical HPLC (5% → 100% B in 5 min, with formic acid): Rt = 2.68 min.

[0078] IVb:HR-ESI MS(ESI):m / z 431.0972, [C47 H 36 N8O3Ru] 2+ Calculated for: 431.0972. Analytical HPLC (5% → 100% B in 5 min, with formic acid): Rt = 2.39 min.

[0079] Phototoxicity evaluation of complex compounds of formula I or formula IV The phototoxicity of several complex compounds of formula I or formula IV was evaluated against several (human and mouse) cancer cell lines and human normal cells under normoxic (21% O2) and hypoxic (2% O2) conditions. The results are shown in Tables 1 to 7 below. In each case, the phototoxicity index (PI) was calculated as follows: PI=IC 50 (Non-irradiated cells under dark conditions) / IC 50 (irradiated cells)

[0080] Cell lines and cell culture conditions. The following cancer cell lines were used: CT-26 (murine colon carcinoma); HT-29 (human colorectal adenocarcinoma); A-549 (human lung adenocarcinoma); and A-2780 (human ovarian carcinoma). The following human normal cells were used: RPE-1 (human retinal pigment epithelium-1), HEK-293 (human fetal kidney), and MRC-5 (human fetal lung fibroblast). Each cell line was cultured in its respective medium and maintained at 37°C in a humidified atmosphere of 5% CO2. The CT-26 cell line was cultured in DMEM medium (Gibco) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin-streptomycin antibiotics (Gibco). The HT-29 cell line was cultured in McCoy's medium (Gibco) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin-streptomycin antibiotics (Gibco). The A-549 cell line was cultured in DMEM / F-12 medium (Gibco) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin-streptomycin antibiotics (Gibco). The A-2780 cell line was cultured in RPMI medium (Gibco) supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin antibiotics (Gibco). The RPE-1 cell line was cultured in DMEM / F-12 medium (Gibco) supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin antibiotics (Gibco). The HEK-293 cell line was cultured in DMEM medium (Gibco) supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin antibiotics (Gibco). The MRC-5 cell line was cultured in DMEM / F-12 medium (Gibco) supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin antibiotics (Gibco).

[0081] Phototoxicity evaluation of the complex compounds of Formula I or Formula IV in 2D monolayer cells under normoxia (21% O). The phototoxicity of test compounds under normoxia (21% O) was evaluated by a fluorescent cell viability assay using resazurin (Acros Organics). Briefly, cells were plated in 96-well plates at 4 × 10 3Cells were seeded in triplicate at a density of 100 cells / well in a final volume of 100 μL per well. After 24 hours, cells were treated with increasing concentrations of metal complexes and control compounds. Test compounds were serially diluted in cell culture medium from a 10 mM or 25 mM stock solution in DMSO, and the solutions were added to give final concentrations ranging from 0.001 μM to 250 μM in a final volume of 100 μL per well. Cells were incubated with the samples in the dark for 4 hours, after which the medium was replaced with fresh medium. To examine the phototoxic effects of the test compounds, an Atlas Photonics LUMOS BIO irradiator was used to irradiate cells at 540 nm (spectral half-width: 32 nm, 40 min, 3.75 mW / cm). 2 , 9.0J / cm 2 ), 645 nm (spectral half-width: 32 nm, 60 min, 2.50 mW / cm 2 , 9.0J / cm 2 ), 670 nm (spectral half-width: 32 nm, 60 min, 3.75 mW / cm 2 , 13.5J / cm 2 ), 740 nm (spectral half-width: 32 nm, 60 min, 3.50 mW / cm 2 , 12.6J / cm 2 ) and 770 nm (spectral half-width: 32 nm, 60 min, 6.75 mV / cm 2 , 24.3J / cm 2 The cells were exposed to light at λ 2000 s. To examine the cytotoxicity of the samples in the dark, the cells were not irradiated. The temperature was then maintained at 37°C throughout the experiment, and a 44-h untreated cell recovery period was allowed. After 44 h of incubation, the medium was aspirated and 100 μL of complete medium containing resazurin (final concentration 0.2 mg / mL) was added. After 4 h of incubation at 37°C, the fluorescent signal of the resazurin product was measured (λ 2000 s.p.m.) using a TECAN Infinite 200 PRO Microplate Reader. ex = 540 nm, λ em = 590 nm). Then, based on the inhibition rate curve, IC 50 Values ​​were calculated. All experiments were performed as three independent trials with triplicate points (n=3 biologically independent replicates) per concentration level.

[0082] Phototoxicity evaluation of the complex compounds of Formula I or Formula IV in 2D monolayer cells under hypoxia (2% O). The phototoxicity of the test compounds under hypoxia (2% O) was evaluated by a fluorescent cell viability assay using resazurin (Acros Organics). Cells were plated in 96-well plates at 2.5 × 10 3 Cells were seeded in triplicate at a density of 100 cells / well in a final volume of 100 μL per well in a 2% O2 incubator. Flasks containing the corresponding cell culture medium were also prepared in a 2% O2 incubator. After 48 h of hypoxia, cells were treated with increasing concentrations of metal complexes and control compounds. Test compounds were serially diluted in medium from 10 mM or 25 mM stock solutions in DMSO, and these solutions were added to the wells at final concentrations ranging from 0.01 μM to 250 μM in a final volume of 100 μL per well. Cells were incubated with the samples in the dark for 4 h, after which the medium was replaced with fresh medium. To examine the phototoxic effects of the samples under hypoxic conditions, a Plas-Labs hypoxia chamber glovebox (856 series) was used. Cells were exposed to light using an Atlas Photonics LUMOS BIO irradiator installed inside the hypoxic chamber (see previous paragraph). To evaluate the cytotoxicity of the samples in the dark, the cells were not irradiated. A 44-hour untreated cell recovery period was then allowed in a 2% O2 incubator, with the temperature maintained at 37°C throughout the experiment. After 44 hours of incubation, the medium was aspirated and 100 μL of complete medium containing resazurin (final concentration 0.2 mg / mL) was added. After 4 hours of incubation in a 37°C, 21% O2 incubator, the fluorescent signal of the resazurin product was measured (λ) using a TECAN Infinite 200 PRO Microplate Reader. ex = 540 nm, λ em = 590 nm). As shown in the previous paragraph, IC 50 values ​​were calculated.

[0083] Phototoxicity results of some complex compounds of formula I or formula IV on cancer cells after irradiation with light of different wavelengths (λ) under normoxic and hypoxic conditions. IC obtained under different conditions 50 The values ​​of the phototoxicity index (PI) and the phototoxicity index (PI) are shown in Tables 1 to 8 below. The uncertainty is the standard error of the mean value of n=3. "nd" means "non determined."

[0084] [Table 1]

[0085] [Table 2]

[0086] [Table 3]

[0087] [Table 4]

[0088] [Table 5]

[0089] [Table 6]

[0090] [Table 7]

[0091] [Table 8]

[0092] Furthermore, it is noteworthy that the complex compound Ic was able to specifically target mitochondria intracellularly (Figures 2 and 3) and inhibit the growth of 3D tumor spheroids (Figure 1). In vivo studies of Ic demonstrated a favorable biodistribution profile and excellent safety profile. As shown in Figure 4, the maximum concentration of the compound in mouse plasma was reached 30 minutes after administration (5 mg / kg), and the compound was completely eliminated after 24 hours. In a maximum tolerated toxicological dose (MTD) study, all parameters, i.e., body weight, body weight gain, food consumption, organ weight, gross necropsy findings, hematological analysis, and biochemical analysis, were comparable to those of the control group in animals (male and female albino Swiss CD1 mice) treated with the two concentrations investigated (10 mg / kg and 30 mg / kg) (Figures 5 and 6). During the observation period (5 days), no deaths or clinical signs were recorded in the male and female experimental groups treated with vehicle and Ic. Furthermore, complex compound Ic was able to reduce tumor growth in a mouse subcutaneous colorectal cancer model (Figure 7).

Claims

1. Compounds of Formula I, including any stereoisomers or E / Z isomers 【Chemistry 1】 (In the formula, M is Fe ２＋ , Ru ２＋ , Os ２＋ , Co ３＋ , Rh ２＋ , Rh ３＋ , Ir ３＋ , Ni ２＋ , Pd ２＋ , Pt ２＋ , and Pt ４＋ is a metal cation selected from the group consisting of y=1, 2 or 3, and z=0, 1 or 2, provided that M is Ni ２＋ , Pd ２＋ or Pt ２＋ When y+z=2, M is Fe ２＋ , Ru ２＋ , Os ２＋ , Co ３＋ , Rh ２＋ , Rh ３＋ , Ir ３＋ , or Pt ４＋ then y+z=3, m=0, 1, 2, 3 or 4; The groups T and T′ are H and (C １ ~C ３ )-alkyl, Base P ３ , P ５ , P ６ , Q ３ , Q ４ , Q ５ , Q ６ , R ３ , R ５ , R ６ , and R ８ are respectively H, (C １ ~C ３ )-alkyl, (C ３ ~C ６ )-cycloalkyl, CH═CHR, CF ３ , CHF ２ , C.H. ２ F, CF ２ CF ３ , F, Cl, Br, I, OR, C(═O)OR, O(C═O)R, C(═O)NR, NRC(═O)R′, NRR′, phenyl, monosubstituted phenyl, disubstituted phenyl, and trisubstituted phenyl, wherein the substituents on the phenyl ring are attached at any of the possible substitution positions; and F, Cl, Br, I, NO ２ , (C １ ~C ３ )-alkyl, OH, O[(C １ ~C ３ )-alkyl], NH ２ , NH[(C １ ~C ３ )-alkyl], and N[(C １ ~C ３ )-alkyl] ２ where m≧1 and the group R ３ is CH ２ When T′=CH ２ Provided that ３ CH ２ and the T′ group closest to the coumarin skeleton, resulting in a compound of formula IV: group R ７ H, NO ２ ,OR,NRR',N(CH ２ COOH) ２ , N(CH ２ CH ２ SO ３ H) ２ , N(CH ２ CONHCH ２ CH ２ NMe ２ ) ２ or the heterocyclic groups 1-aziridinyl, 1-azetidinyl, 1-pyrrolidinyl, 1-piperidinyl, 4-morpholinyl, or 1-piperazinyl, respectively, unsubstituted, mono-(R)-substituted, di-(R,R')-substituted, or tri-(R,R',R'')-substituted; alternatively, R ６ , R ７ and R ８ together form a bicyclic ring system of the formula: １ , V ２ , V ３ , V ４ , V ５ , V ６ , V ７ , V ８ , V ９ , V １０ , V １１ and V １２ are H and (C １ ~C ３ )-alkyl), 【Chemistry 2】 group R ４ is H, (C １ ~C ３ )-alkyl, (C ３ ~C ６ )-cycloalkyl, CF ３ , CHF ２ , C.H. ２ F, CF ２ CF ３ , F, Cl, Br, I, OR, C(═O)OR, O(C═O)R, C(═O)NR, NRC(═O)R′, NRR′, phenyl, mono-(R)-substituted phenyl, di-(R,R′)-substituted phenyl, tri-(R,R′,R″)-substituted phenyl, or the group CE １ E ２ E ３ where E １ and E ２ are H, F, Cl, Br, I, OR, NRR', NO ２ , (C １ ~C ３ is a group independently selected from the group consisting of mono-(R)-alkyl, phenyl, and mono-(R)-substituted phenyl; ３ is OH, O[(C １ ~C ３ )-alkyl], or a group of the formula: １ and D ２ At each occurrence, H, O[CH ２ ] ｑ -X, S [CH ２ ] ｑ -X, and N[[CH ２ ] ｑ -X] ２ wherein q=1 or 2 and X=F, Cl, Br or I; 【Transformation 3】 or a group of the following formula: ３ is H, H(C=O), [(C １ ~C ３ )-alkyl](C═O), (OH) ２ P(=O)O-CH ２ -O(C=O), or (PhO)(OH)P(=O) 【Chemistry 4】 and Lig, at each occurrence, is a bidentate ligand independently selected from the group consisting of ligands of the following formula: Lig1, Lig2, Lig3, Lig4, Lig5, Lig6, Lig7, Lig8, Lig9, Lig10, Lig11, Lig12, and Lig13, wherein G is a N atom or a H atom in the formation of a carbon-metal bond. ＋ is either a C atom with one negative charge formed in situ from C—H when R １０ , R １１ , R １２ , R １３ , R １４ , R １５ , R １６ , R １７ , R １８ , R １９ , R ２０ and R ２１ are respectively H, (C １ ~C ３ )-a group independently selected from the group consisting of alkyl, phenyl, and mono-, di-, and tri-substituted phenyl, wherein the substituents attached to the phenyl ring are independently selected from F, Cl, Br, I, OR, and NRR'; 【Transformation 5】 【change】 【change】 A is an anion derived from a pharmaceutically acceptable acid, n is an integer or fraction such that the compound of formula I is electrically neutral; where R, R', and R'' are H, F, Cl, Br, I, NO, respectively, in each occurrence above. ２ , (C １ ~C ３ )-alkyl, OH, O[(C １ ~C ３ )-alkyl], NH ２ , NH[(C １ ~C ３ )-alkyl], and N[(C １ ~C ３ )-alkyl] ２ and R is a group independently selected from the group consisting of:

2. The anion A is Cl － ,Br － , P.F. ６ － , P.F. ４ － , B.F. ４ － , ClO ４ － , C.F. ３ SO ３ － , S.O. ４ ２－ , C.F. ３ COO － 2. The compound of claim 1, wherein the compound is selected from the group consisting of acetic acid, formic acid, and oxalic acid.

3. The anion A is Cl － 3. The compound of claim 2, wherein:

4. y=1, z=2, and the metal cation M is Fe ２＋ , Ru ２＋ , Os ２＋ , Co ３＋ , Rh ２＋ , Rh ３＋ , Ir ３＋ , or Pt ４＋ The compound according to any one of claims 1 to 3,

5. the bidentate ligand Lig is Lig1 or Lig2, and the group R １０ , R １１ , R １２ , R １３ , R １４ , R １５ , R １６ , R １７ , R １８ , R １９ , R ２０ and R ２１ 5. The compound of any one of claims 1 to 4, wherein each of: at each occurrence is independently selected from the group consisting of H, methyl, and phenyl.

6. The compound of any one of claims 1 to 5, wherein G is N.

7. The compound according to claim 6, wherein the bidentate ligand Lig is 2,2'-bipyridine (bpy) or 4,7-diphenyl-1,10-phenanthroline (BPhen).

8. The group P ３ , P ５ , P ６ , Q ３ , Q ４ , Q ５ , Q ６ , R ３ , R ５ , R ６ and R ８ Each of these, at each occurrence, is １ ~C ３ )-alkyl, and the group R ７ H, N [(C １ ~C ３ )-alkyl)] ２ Or NO ２ and alternatively, the group R ６ , R ７ and R ８ together form a bicyclic ring system of the formula shown in claim 1, V １ , V ２ , V ３ , V ４ , V ５ , V ６ , V ７ , V ８ , V ９ , V １０ , V １１ and V １２ The compound of any one of claims 1 to 7, wherein each of is H.

9. The group R ４ But, H, (C １ ~C ３ )-alkyl, CF ３ , CH(CH ３ ) OH, CH(CH ３ )O(C=O)-[CH ２ ] ３ -Ph, or a group of the following formula: 【Transformation 6】 Or, a compound of the formula: CH(CH ３ )O(C=O)-[CH ２ ] ２ —(C═O)—CH ２ -NH ２ The compound according to any one of claims 1 to 8, wherein the group is

10. The compound according to any one of claims 1 to 9, wherein m=0, 1 or 2.

11. The metal cation M is Ru ２＋ The compound according to any one of claims 1 to 10,

12. Formula I is represented by the following formulae Ia, Ib, Ic, Id, Ie, If, Ig, Ih, Im, In, Io, Ip, Iq, IVa and IVb: 【Transformation 7】 【change】 【change】 12. The compound of claim 11 selected from the group consisting of:

13. Compounds of formula II, including any stereoisomers or E / Z isomers: 【Transformation 8】 (In the formula, m, P ３ , P ５ , P ６ , Q ３ , Q ４ , Q ５ , Q ６ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , T and T' are as defined in any one of claims 1, 8, 9 and 10).

14. Formula II is selected from the group consisting of the following formulae IIa, IIb, IIc, IId, IIe, IIf, IIm, IIo, IIn, IIp, IIq, IIr, Va, and Vb: 【Chemistry 9】 【change】 【change】 14. The compound of claim 13 selected from the group consisting of:

15. A pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I as defined in any one of claims 1 to 12 together with a suitable amount of an excipient, carrier or vehicle.

16. A compound of formula I as defined in any one of claims 1 to 12 for use in human therapy.

17. A compound of formula I as defined in any one of claims 1 to 12 for use as a photosensitizer in the photodynamic therapy of human diseases.

18. 18. The compound of claim 17, wherein the human disease is cancer, certain skin conditions, fungal infections, or microbial infections.

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