Medical compounds

Abstract

The present invention provides a medicinal compound comprising a synthetic flavonoid derivative according to formula (I), which has a group at the C-3 position: wherein at least two of R2-R6 are H, and the rest are independently selected from: H, OH, R1, OR1, NO2, NH2, NHR1, F, Cl, Br, I; wherein R1 is a free radical. The present invention also provides dimerization, trimerization and tetramerization compounds, wherein two or more quercetin units are connected to a central linker (skeleton) via position 3 and a spacer. The medicinal compound can be used to treat malignant tumors, in particular malignant epithelial tumors, such as colon cancer, melanoma and / or squamous cell carcinoma of the skin, lung cancer, breast cancer, basal cell carcinoma (basal cell tumor). #imgabs0#

Description

[0001] The present invention relates to a medicinal compound of the type according to the preamble of the first claim, in particular for use in the treatment of malignant tumors, more particularly for use in the treatment of malignant epithelial tumors and lung cancer, colon cancer, melanoma and squamous cell carcinoma of the skin, or for anti-inflammatory and analgesic activity.

[0002] In recent decades, neoplastic diseases are becoming one of the greatest challenges in medicine and one of the main causes of death, both in developing and industrialized countries. About 1.6 million new cases of neoplasia are diagnosed each year, and more than 100 different types of neoplasia are known to date, including a wide variety of epithelial tumors, especially breast cancer, lung cancer, colorectal cancer, prostate cancer, liver cancer and skin cancer.

[0003] Despite the progress achieved in the treatment of neoplastic diseases, further advances are undoubtedly needed, mainly due to the severe side effects associated with the most common anti-cancer therapies, such as chemotherapy and radiotherapy.

[0004] In the search for new drugs, several studies have attempted to investigate the antitumor potential of substances derived from fruits and vegetables, known as phytoprotectants. Among the phytoprotectants known to date, flavonoids and flavonoids, especially quercetin, have shown antitumor properties in many studies against different tumor cell lines, such as leukemia HL-60, colon cancer SW-480, mouse breast cancer 4T1 and squamous cell carcinoma A431.

[0005] Quercetin (3,3',4',5,7-pentahydroxyflavone) is a natural flavonoid characterized by the presence of 5 hydroxyl groups, which is responsible for its biological activity. It is found in a variety of plants, especially broccoli, tea, onion, carrot. In nature, its activity can inhibit bilateral growth of embryos in plants. The most representative quercetin derivatives are glycosides and esters. Numerous studies have shown that quercetin has antioxidant capacity, which is attributed to the presence of hydroxyl groups in the A ring and catechol groups in the B ring. These properties allow quercetin to remove oxygen free radicals by transferring hydrogen or electrons or chelating metal ions, thereby inhibiting pro-inflammatory enzyme activities.

[0006] Flavonoid compounds inhibit lipid peroxidase activity by preventing LDL (low-density lipoprotein) oxidation and cell membrane damage. In addition, flavonoids increase glutathione levels and prevent the formation of free radicals.

[0007] The therapeutic efficacy of quercetin is closely related to its bioavailability after oral administration. After ingestion, quercetin is hydrolyzed by intestinal glycosidases and absorbed through the stomach or intestines as aglycones. Subsequent enzymatic modification of the molecule converts it into pharmacologically active forms such as quercetin-3-glycoside, quercetin-3-sulfate, quercetin-3-glucuronide, and quercetin-3-methyl ether.

[0008] Quercetin is metabolized very quickly in the human body, so the availability of its active form is limited. In addition, quercetin is poorly water soluble, which further affects its therapeutic potential. Therefore, it is necessary to increase the water solubility of quercetin and delay metabolism to obtain prolonged blood and tissue levels of the molecule.

[0009] The anti-tumor effect of quercetin involves multiple aspects:

[0010] 1) Cell growth inhibition: Quercetin has shown antiproliferative effects on different types of neoplasia both in vitro and in vivo. In vitro, it inhibits cell growth of L1210 and P388 leukemia cell lines; breast tumors; COLO 20DM colon cancer; OVCA 43 ovarian cancer and A431 squamous cell carcinoma. The molecular mechanism involved appears to be the regulation of pro-proliferative intracellular cascades, such as PI3K / akt, Her-2 / neu, Wnt / β-catenin and EGFR. Quercetin appears to inhibit the activity of mammalian target of rapamycin (mTOR), which is overactive in tumor cells and is crucial in controlling intracellular growth signals, apoptosis mechanisms, protein synthesis and PI3K / Akt activation. In addition to inhibiting the activation of PI3K / Akt, quercetin also downregulates Her-2 / neu (human epidermal growth factor receptor 2), which is overexpressed in certain breast tumors, and allows inhibition of the Wn / β-catenin signaling pathway, resulting in reduced cell growth. In addition to modulating the above-mentioned pro-proliferative intracellular signaling pathways, quercetin also interferes with normal cell cycle progression, resulting in reduced cell growth.

[0011] 2) Inhibition of the metastatic process: The metastatic process is closely associated with the production of metalloproteinases (MMPs), which are enzymes responsible for the degradation of the extracellular matrix and the invasion of tumor cells into surrounding tissues. Several studies have analyzed the potential of quercetin to inhibit MMPs.

[0012] 3) Induce apoptosis: Apoptosis, or programmed cell death, is essential for the body’s homeostasis. Dysregulation and defects in the apoptotic process lead to tumor development. Quercetin has been shown to increase the synthesis of pro-apoptotic factors in breast and colorectal cancer cell lines.

[0013] Despite these encouraging aspects, flavonoids and flavonoids are currently subject to several limitations as potential candidates for the treatment of malignancies. Indeed, as mentioned previously, these molecules have low solubility in H2O (1 mcg / mL), low intrinsic activity, low absorption (<10%), rapid metabolism (<1 hr), rapid clearance from the body (>40%), and multiple inactive metabolites.

[0014] These characteristics indicate that it is highly necessary to identify quercetin derivatives with higher bioavailability and bioactivity in order to apply these plant protection products in the treatment of malignant tumors.

[0015] In this context, the technical task of the present invention is to design a medicinal compound, in particular for the treatment of malignant tumors, in particular epithelial tumors, or for anti-inflammatory activity, etc., which is able to substantially eliminate at least some of the above-mentioned disadvantages.

[0016] Within the scope of said technical task, the main purpose of the present invention is to obtain a medicinal compound which is used in the treatment of malignant tumors in general, in particular lung cancer, colon cancer, melanoma, squamous cell carcinoma of the skin and basal cell carcinoma (basal cell tumor).

[0017] The technical task and the specific objects are achieved by the medicinal compounds claimed in the attached claim 1.

[0018] The features and advantages of the present invention will become more apparent from the detailed description of preferred embodiments of the present invention with reference to the accompanying drawings, in which:

[0019] Figure 1a , 1b , 2a, 2b, 3a, 3b, 4a, 4b, 5a, 5b, 6a, 6b, 7a, 7b, 8a, 8b, 9a, 9b, 10a, 10b, 11a, 11b, 12, 13a, 13b, 14, 15, 16, 17, 18, 19, 20, 21, 22a, 22b show the test results of the following treatments with standard compounds and with the medicinal compounds according to the present invention.

[0020] Preferred embodiments are described in the dependent claims.

[0021] In this document, measurements, values, shapes and geometric references (such as perpendicularity and parallelism) associated with terms such as "about" or other similar terms such as "almost" or "substantially" should be understood to mean that unless there is a measurement error or inaccuracy due to production and / or manufacturing defects, it is especially slightly different from the value, measure, shape or geometric reference to which it is associated. For example, these terms, if associated with a value, preferably represent a difference of no more than 10% of the value itself.

[0022] Furthermore, when used, terms such as “first,” “second,” “higher,” “lower,” “primary,” and “secondary” do not necessarily indicate an order, priority relationship, or relative position, but may simply be used to more clearly distinguish different components from each other.

[0023] Unless otherwise stated, the measurements and data reported herein should be considered to have been performed according to the International Standard Atmosphere ICAO (ISO 2533).

[0024] The medicinal compounds according to the invention are particularly useful for the treatment of malignant tumors, in particular malignant epithelial tumors. The term "epithelial tumor" is intended to mean a malignant tumor originating from epithelial tissue (skin, mucous membranes, glandular system). Neoplastic transformation alters the numerical homeostasis of cells in tissues, leading to uncontrolled growth and genomic instability.

[0025] In particular, the compounds according to the invention are used in the treatment of one or more of the following malignancies, in particular malignant epithelial tumors, colon cancer, melanoma and / or squamous cell carcinoma of the skin, lung cancer, breast cancer, basal cell carcinoma (basal cell tumor).

[0026] In addition, the compound according to the invention is preferably used to prevent the proliferation of the above-mentioned tumors, because it affects the proliferation capacity of the tumor and apoptosis. It can also induce cell death and apoptosis, for example by increasing ROS (reactive oxygen species), disrupting mitochondrial activity, damaging DNA or activating caspase 3, 8 and 9 and causing cell cycle arrest.

[0027] In addition, the compounds of the present invention are useful in one or more of the following malignancies: head and neck cancer, stomach cancer, liver cancer, pancreatic cancer, breast cancer, prostate cancer, bladder cancer, kidney cancer, mesothelioma, and ovarian cancer.

[0028] The compounds according to the invention are not suitable for the treatment of hepatitis C and the consequent malignant liver tumors or hepatocellular carcinomas caused by hepatitis C virus infection.

[0029] Furthermore, the compounds according to the invention are useful in one or more of the following malignancies: salivary gland tumors, esophageal cancer, small intestinal tumors, gallbladder cancer and extrahepatic bile duct cancer, soft tissue sarcomas, cervical cancer, uterine cancer, testicular cancer, urinary tract tumors, choroidal melanoma, thyroid cancer, endometrial cancer, retinal tumors, uveal melanoma, anal canal cancer and anal cancer, bone and joint cancer.

[0030] Finally, the compounds according to the invention are used in non-tumor therapy, i.e. the use of our compounds is:

[0031] - anti-inflammatory activity,

[0032] - analgesic activity,

[0033] - antimicrobial activity (bacteria, viruses, fungi),

[0034] -Immunomodulatory activity

[0035] - Antioxidant activity,

[0036] -Cellular protective and nourishing activity,

[0037] - Anti-degeneration and anti-aging activity,

[0038] - idiopathic chronic inflammatory diseases (Crohn's disease, ulcerative colitis, irritable bowel syndrome, fibromyalgia, arthritis, myopathy),

[0039] - Neurodegenerative diseases (Alzheimer's disease, Huntington's disease, ALS, Parkinson's disease, senile dementia, glaucoma),

[0040] - Syndrome due to long-term toxicity of chemotherapy and radiotherapy (central and peripheral neurotoxicity, inflammation, oxidative stress, fatigue, vomiting, nausea, diarrhea, constipation, impotence, hearing loss, heart disease).

[0041] The compounds according to the invention are preferably synthetic flavonoid derivatives according to formula (I):

[0042]

[0043] It has a group at the C-3 position,

[0044] wherein at least two of R2-R6 are H, and the others are independently selected from: H, OH, R1, OR1, NO2, NH2, NHR1, F, Cl, Br, I, and more particularly, R2, R3 and R6 are H, and R4 and R5 are OH,

[0045] Wherein R1 is a moiety preferably selected from the following:

[0046] -H;

[0047] -C 1-24 Alkyl or heteroalkyl, C 1-24 Alkenyl or heteroalkenyl, C 1-24 Alkynyl or heteroalkynyl;

[0048] - acyl residues of fatty acids. The fatty acids may be saturated, unsaturated or polyunsaturated of synthetic or natural origin.

[0049] Preferably, R1 is:

[0050]

[0051] wherein R'1 is selected from the same compound as R1,

[0052] Where a, b and c are each 0 to 12,

[0053] Wherein X, Y and Z are each independently selected from: CH2, O, N(R1), S, NH, SO, SO2, OC(O), CO, NHC(O), C(O)NH, NH-C(O)-NH, NH-C(S)-NH.

[0054] Regarding the synthesis of quercetin ethers 1, 2 and 3, as reported by Rolando et al., this synthetic strategy provides orthogonal protection of quercetin (Tetrahedron Letters, 2011, 52, 4738). The synthesis of quercetin ethers 1, 2 and 3 starting from commercially available rutin 6 provides extensive benzylation of free hydroxyl groups, followed by selective hydrolysis of disaccharides at position 3 and subsequent alkylation with appropriate alkyl bromides, and finally removal of benzyl groups by hydrogenation catalyzed by Pd / C. The strategy is simplified as follows:

[0055]

[0056] The synthesis of quercetin esters 4 and 5 was carried out by an enzymatic strategy reported by D. Lambusta et al. (J. Mol. Catal. B-Enzymatic, 2003, 22, 271). The strategy includes esterification of quercetin 9 on all five hydroxyl groups, followed by selective alcoholysis mediated by appropriately selected lipases from Candida antarctica (CAL) and Mucor miehei (MML), as shown in the following scheme:

[0057]

[0058] Similar to the synthesis of product 8, the synthesis of the product of series 11 is shown below, as shown in the following scheme:

[0059]

[0060] The compounds according to the invention also include dimer, trimer and tetramer molecules, in which two or more quercetin units are linked to a suitable central linker (backbone) via the 3-position and a spacer, as described below:

[0061]

[0062] To investigate the effects caused by flexibility and distance between quercetin units, both spacers and central backbones were selected.

[0063] The first set of dimer molecules was synthesized by the strategy shown in the following scheme, where the ethyl ester of intermediate 11a was hydrolyzed by treatment with sodium hydroxide in THF / H2O, and the corresponding EDC / HOBt-activated carboxylic acid 12 was condensed with the appropriate diamine (H2N-linker-NH2) 13. Finally, extensive debenzylation of intermediate 14 was performed by classical catalytic hydrogenation to obtain the final product 15:

[0064]

[0065] The second set of dimer molecules was synthesized by the strategy shown in the following scheme, which involves the removal of the protecting group (BOC) from amine 11b by treatment with 4N HCl in THF / H2O. The corresponding amine 16 was then condensed with the appropriate dicarboxylic acid molecule 17 under EDC / HOBt activation. Finally, extensive debenzylation of intermediate 18 was performed by catalytic hydrogenation to obtain the final product 19.

[0066]

[0067] Similarly, the trimer molecule was synthesized starting from intermediate 16 by condensation with an appropriately selected tricarboxylic acid molecule 20 under EDC / HOBt activation (as shown in the scheme below). Extensive debenzylation of intermediate 21 was performed by classical catalytic hydrogenation to obtain the final product 22.

[0068]

[0069] The tetrameric molecule was synthesized starting from the intermediate 16 by condensation with an appropriately selected tetracarboxylic acid molecule 23 under EDC / HOBt activation (as shown in the scheme below).

[0070]

[0071] Finally, extensive debenzylation of intermediate 24 was carried out by classical catalytic hydrogenation to afford the final product 25 (as shown in the following scheme).

[0072]

[0073] Other compounds according to the invention include molecules in which the pharmacophore appendage is linked to the oxygen at position 3 of quercetin via an appropriate spacer bridge (as shown in the figure below).

[0074]

[0075] [“Spaziatore” = spacer;

[0076] "Farmacoforo" = pharmacophore]

[0077] These molecules were synthesized starting from compound 11(bc) according to the following scheme, thereby obtaining quercetin derivatives 26 (obtained by complete debenzylation of 11c) and 16 (obtained by removal of the protecting group Boc from 11b in an acidic environment):

[0078]

[0079] As pharmacophore groups, some compounds according to the present invention exhibit chemical structures related to endocannabinoids and N-acylethanolamines (NAE; the structure of the biological compound related to humans is shown in the figure below).

[0080]

[0081] [“2-arachidonilglicerolo” = 2-arachidonylglycerol;

[0082] "2-arachidonil gliceril etere" = 2-arachidonyl glycerol ether;

[0083] "Virodamina" = O-arachidonoylethanolamine (Virodamine);

[0084] "N-arachidonoildopamina" = N-arachidonyl dopamine;

[0085] "Palmitoiletanolammide" = palmitoylethanolamide]

[0086] For example, among the main structures listed in the figure above, mention can be made of palmitic ethanolamide (PEA), anandamide (cannabinoids) and dopamineamide with anandamide (N-arachidonyl dopamine). By comparing these structures with the structure of the quercetin derivative 27 (shown in the scheme below), it seems that in the latter, the chain connected to the oxygen at position 3 can mimic the ethanolamine part of endocannabinoids and NAE in its final aminoethanol part, while the innermost ethylene part acts as a spacer between quercetin and the pharmacophore (for example in the figure below, the upper figure shows PEA; the lower figure shows that by linking the quercetin derivative 27 with a suitable acyl group, a compound comprising the PEA structure according to the invention is formed, and the two structures are separated by an ethylene bridge between the two oxygens).

[0087]

[0088] [“Palmitoile” = palmitoyl]

[0089] Thus, by attaching an appropriate acyl moiety to quercetin derivative 27, a quercetin molecule was obtained, the oxygen at position 3 of which was attached to an endocannabinoid or NAE via an ethylene spacer. The quercetin derivative 27 obtained by debenzylation of compound 16 was directed to a parallel synthesis to obtain an amide via its amino function, each time reacting it with another acid (28(ag)) previously activated with EDC and HOBt to obtain the amide 29(ag) (as shown in the following scheme).

[0090]

[0091] ["Ac.Palmitico" = palmitic acid;

[0092] "Ac.Docosanoico" = docosanoic acid;

[0093] "DMF anidra" = anhydrous DMF;

[0094] "ta" = rt;

[0095] "Ac.Oleico" = oleic acid;

[0096] "Ac.Linoleico" = linoleic acid;

[0097] "Ac.Arachidonico" = arachidonic acid;

[0098] "Ac.Docosaesaenoico" = Docosahexaenoic acid;

[0099] "Ac.4"-Metossibifenil-4-Carbossilico" = 4"-Methoxybiphenyl-4-carboxylic acid]

[0100] Thus, in addition to the substances containing PEA (29a) and cannabinoids (29e), compounds according to the invention with different, unique and novel acyl residues were synthesized (compounds 29b, 29c, 29d, 29f), thereby also obtaining information about the importance of the type of fatty acid in the molecule. A molecule (29g) was also prepared in which the acyl substituent was no longer a long-chain fatty acid, but a diphenyl derivative, in order to obtain information about the effect and biological activity of the transition from a linear polyunsaturated system to an aromatic system and to evaluate it.

[0101] Furthermore, compounds according to the invention include quercetin-NAE hybrid structures, such as heterotrimeric molecules of the Q2E type, i.e. containing two quercetin units (Q) and one endocannabinoid or NAE unit (E). For this purpose, the endocannabinoid anandamide ( FIG. 3 ) (wherein two dopamine phenolic groups allow the connection of two quercetin units) was used as a model. Molecule 36, in which anandamide (present in the reference template, i.e. the endocannabinoid anandamide) was replaced by palmitic acid and each Q2E quercetin was bridged to a different dopamine hydroxyl group (as shown in the following scheme), was prepared as a first representative of this type of compound.

[0102]

[0103] [“DMF anidra” = anhydrous DMF;

[0104] "ta" = rt]

[0105] Briefly, dopamine 30 was reacted with palmitic acid activated by EDC / HOBt to produce amide 31, which was subsequently converted to diester 33 by reaction with bromide 32. Alkaline hydrolysis of the diester gave diacid 34, which, under EDC and HOBt activation, reacted with excess quercetin tetrabenzyl ester 16 to obtain compound 35, which was fully reductively debenzylated by hydrogenation in the presence of Pd / C to afford the target molecule 36. Thus, a molecule was obtained in which an artificial analog of the endocannabinoid anandamide dopamine, palmitoyl dopamine, was linked to two quercetin units to form a Q2E-type heterotrimer.

[0106] The synthesis examples of various components are listed below. All chemical reagents used were analytically pure and used as received.

[0107] In Bruker Avance 400 TM The spectrometer recorded the indicated deuterated solvent at 400.13 and 100.62 MHz respectively. 1 H and 13 C-NMR experiments. Coupling constants are reported in Hertz and are rounded to the nearest 0.1 Hz. When necessary, chromatographic purification was performed by flash chromatography on silica gel (70-230 mesh) using the specified eluents.

[0108] When specified, reactions were performed using a CEM Discover microwave reactor.

[0109] Synthesis example of 5,7-bis(benzyloxy)-2-(3,4-bis(benzyloxy)phenyl)-3-hydroxy-4H-benzopyran-4-one 7:

[0110] Anhydrous K2CO3 (1.81 g; 13.1 mmol) and benzyl bromide (2.24 g; 13.1 mmol) were added sequentially to a solution of 2-(3,4-dihydroxyphenyl)-4,5-dihydroxy-3-[3,4,5-trihydroxy-6-[(3,4,5-trihydroxy-6-methyl-oxa-2-yl)oxymethyl]oxa-2-yl]oxy-benzopyran-7-one 6 in DMF (12 ml). The reaction mixture was stirred at room temperature under an argon atmosphere. After completion (about 24 hours), the reaction mixture was diluted with EtOAc (40 ml) and washed with H2O (2×30 ml). The organic phase was dried over anhydrous Na2SO4, filtered, and the solvent was removed under reduced pressure. A mixture of HCl (37%) / MeOH=2 / 98 v / v was added to the residue and heated to reflux (T°=65°C) for 2 hours.

[0111] After completion, the mixture was cooled to room temperature and then filtered through a Buchner funnel and the orange solid was finally washed with cold methanol. The final product 7 thus obtained did not require further final purification (450 mg; 42.45%).

[0112]

[0113] 1 H NMR (400MHz, CDCl3): 7.88 (s, 1H), 7.75 (d, 1H, J = 6.5Hz), 7.65-7.25 (m, 21H, Ph), 7.01 (d, 2H), 6.56 (s, 1H ,J=8.7Hz),6.45(s,1H),5.26(s,2H,CH2Ph),5.23(s,2H,CH2Ph),5.21(s,2H,CH2Ph),5.11(s,2H,CH2Ph);

[0114] 13 C NMR(100MHz, CDCl3):171.70,163.19,159.33,158.63,150.14,148.58,141.83,137.68,137.13,136.82,136.15,135.59,128.74,128.62,128.5 3,128.45,127.89,127.85,127.77,127.62,127.53,127.18,126.63,124 .26,121.20,114.17,106.68,97.52,93.66,71.52,70.93,70.67,70.53.

[0115] Synthesis example of 5,7-bis(benzyloxy)-2-(3,4-bis(benzyloxy)phenyl)-3-ethoxy-4H-benzopyran-4-one 8a:

[0116] Ethyl iodide (0.072 ml; 0.0009 mol) and anhydrous K2CO3 (0.104 g; 0.0007 mol) were added successively to a solution of 5,7-bis(benzyloxy)-2-(3,4-bis(benzyloxy)phenyl)-3-hydroxy-4H-benzopyran-4-one 7 (0.2 g; 0.0003 mol) in anhydrous DMF (2.14 ml). The reaction mixture was stirred at a temperature of 65° C. for 12 hours. After completion, the reaction was partitioned between water (10 ml) and EtOAc / DCM (1 / 1; 30 ml). The organic phase was washed with saturated aqueous NaCl solution and the aqueous phase was extracted with 10 ml EtOAc / DCM (1 / 1). The combined organic phases were dried over anhydrous sodium sulfate, the solvent was removed under reduced pressure, and purified by flash column chromatography (SiO2; EtOAc / n-hexane 25%) to give a residue (290 mg) in the form of a yellow solid. The product 8a was isolated as a white solid (120 mg, 60%).

[0117]

[0118] 1 H NMR(400MHz, CDCl3):7.83(d,2H),7.68(dd,1H),7.61(d,2H),7.54-7.27(m;19H),7.02 (d,1H),6.53(d,1H),6.45(d,1H),5.24(d,6H),5.09(s,1H),4.06(q,2H),1.26(t,3H);

[0119] 13 C NMR (100MHz, CDCl3):173.95,162.66,159.77,158.65,152.70,150.65,148 .27,140.23,137.05,136.77,136.40,135.71,128.74,128.57,128.53,128. 41,127.95,127.87,127.61,127.58,127.31,127.19,126.66,124.15,122.20,115.26,113.81,98.01,93.86,71.43,70.93,70.79,70.45,68.15,15.60.

[0120] Synthesis example of 5,7-bis(benzyloxy)-2-(3,4-bis(benzyloxy)phenyl)-3-hexyloxy-4H-benzopyran-4-one 8b:

[0121] Hexyl bromide (0.084 ml; 0.0006 mol), anhydrous K2CO3 (0.104 g; 0.0007 mol) and KI (0.015 g; 0.00009 mol) were added sequentially to a solution of 5,7-bis(benzyloxy)-2-(3,4-bis(benzyloxy)phenyl)-3-hydroxy-4H-benzopyran-4-one 7 (0.2 g; 0.0003 mol) in anhydrous DMF (2.14 ml). The reaction mixture was stirred at a temperature of 65°C for 12 hours. After completion, the mixture was partitioned between water (10 ml) and EtOAc / DCM (1 / 1; 30 ml). The organic phase was washed with saturated aqueous NaCl solution and the aqueous phase was extracted with 10 ml EtOAc / DCM (1 / 1). The combined organic phases were dried over anhydrous Na2SO4 and the solvent was removed under reduced pressure. Purification by cryoprecipitation from EtOAc / hexane solution gave a yellow oily residue (173 mg).The white solid product corresponding to 8b was isolated from the liquid by centrifugation and traces of solvent were removed under reduced pressure.

[0122] White solid 8b (67.5 mg; 30%).

[0123]

[0124] 1 H NMR (400MHz, CDCl3): 7.82 (d, 2H, J = 1.8Hz), 7.68 (dd, 1H, J1 = 8.6Hz; J2 = 1.8, Hz), 7.62 (d, 2H, J = 7.6Hz, CH2Ph), 7.55-7.27 (m, 18H, CH2Ph), 7.02 ( d,1H,J=8.6Hz),6.53(d,1H,J=2.0Hz),6.45(d,1H,J=2.0Hz),4.02(t,2H ,J=7.1Hz),1.56-1.53(m,2H),1.40-1.20(m,6H),0.87(t,3H,J=7.1Hz).

[0125] 13C NMR (100MHz, CDCl3):173.82,162.56,159.68,152.53,150.60,148.24,1 40.43,136.73,136.40,128.66,128.45,127.87,127.80,127.53,127.28 ,127.12,126.59,124.07,122.28,115.23,113.72,110.01,97.96,93.79 ,72.67,71.40,70.85,70.68,70.37,31.59,30.08,25.52,22.54,13.98.

[0126] Synthesis example of 5,7-bis(benzyloxy)-2-(3,4-bis(benzyloxy)phenyl)-3-hexadecyloxy-4H-benzopyran-4-one 8c:

[0127] Hexyldecyl bromide (0.2 g; 0.0007 mol), anhydrous K2CO3 (0.121 g; 0.0009 mol) and KI (0.017 g; 0.0001 mol) were added sequentially to a solution of 5,7-bis(benzyloxy)-2-(3,4-bis(benzyloxy)phenyl)-3-hydroxy-4H-benzopyran-4-one 7 (0.2 g; 0.0003 mol) in anhydrous DMF (3 ml). The reaction mixture was stirred at a temperature of 65°C for 2 hours. After completion, the reaction mixture was partitioned between water (10 ml) and EtOAc / DCM (1 / 1; 30 ml). The organic phase was washed with saturated aqueous NaCl solution and the aqueous phase was extracted with 10 ml EtOAc / DCM (1 / 1). The combined organic phases were dried over anhydrous Na2SO4 and the solvent was removed under reduced pressure. The obtained residue (490 mg) was purified by flash column chromatography (SiO2; EtOAc / n-hexane 15%) to isolate the desired product 8c (250 mg, 73.5%) in the form of a white solid.

[0128]

[0129] 1H NMR (400MHz, CDCl3): 7.79 (d, 1H, J = 1.8Hz), 7.66 (dd, 1H, J1 = 8.6Hz, J2 = 1.8Hz), 7.59 (d, 2 H,J=7.4Hz,CH2Ph),7.53-7.28(m,18H,CH2Ph),7.01(d,1H,J=8.6Hz),6.52(d,1H,J=1.8H z),6.44(d,1H,J=1.8Hz),5.254(s,3H,CH2Ph),5.248(s,3H,CH2Ph),5.23(s,3H,CH2Ph), 5.09(s,3H,CH2Ph),3.98(t,2H,J=7.1Hz),1.67(t,2H),1.30-1.19(m,29H),0.38(q,3H);

[0130] 13 C NMR (100MHz, CDCl3):173.76,162.54,159.65,158.52,152.44,150.60,148.25,137 .00,136.71,136.40,135.70,128.61,128.45,128.40,128.26,127.83,127.48,127. 26,127.10,126.57,122.23,115.22,113.71,109.99,97.91,93.79,72.65,71.37,70.84,70.65,70.32,31.82,30.13,29.61,29.55,29.42,29.26,25.87,22.59,14.02.

[0131] Synthesis example of compound 11a.

[0132] Ethyl 4-bromobutyrate (218 μl; 1.508 mmol), anhydrous K2CO3 (250 mg; 1.810 mmol) and KI (36 mg; 0.226 mmol) were added successively to a solution of 5,7-bis(benzyloxy)-2-(3,4-bis(benzyloxy)phenyl)-3-hydroxy-4H-benzopyran-4-one 7 (0.5 g; 0.754 mmol) in anhydrous DMF (4 ml). The reaction mixture was stirred at a temperature of 65°C for 12 hours. After completion, the reactants were distributed between water (20 ml) and EtOAc (20 ml). The organic phase was washed with saturated aqueous NaCl solution and the aqueous phase was extracted with 10 ml of EtOAc. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (SiO2; EtOAc / n-hexane 25%). The product 11a was isolated as a white oil (440 mg, 75%).

[0133]

[0134] 1 H NMR (400MHz, DMSO-d6): 7.76 (s, 1H), 7.66 (d, 1H, J = 8.0Hz), 7.61 (d, 2H, J = 7 .9Hz),7.49(m,6H),7.38(m,12H),7.36(d,1H,J=4.8Hz),6.91(d,1H,J=2.0H z),6.69(d,1H,J=2.0Hz),5.23(m,8H),4.01(q,2H,J=6.8Hz),3.91(t,2H,J= 6.4Hz), 2.38 (t, 2H, J = 7.2Hz), 1.85 (t, 2H, J = 6.8Hz), 1.12 (t, 3H, J = 7.2Hz).

[0135] 13 C NMR(100MHz,DMSO-d6):173.4,173.1,163.5,160.0,159.0,152.8,151.2,148.7,140.4,138.0,137.8,137.7,137.0,129.5,129.4, 129.1,129.0,128.9,128.5,128.4,127.9,124.0,122.9,115.1,114.7,109.9,98.7,95.1,71.6,71.4,71.0,70.9,30.9,25.9,15.0.

[0136] Synthesis example of compound 11b.

[0137] 2-[2-(Boc-amino)ethoxy]ethyl bromide (404 mg; 1.508 mmol), anhydrous K2CO3 (250 mg; 1.810 mmol) and KI (36 mg; 0.226 mmol) were added successively to a solution of 5,7-bis(benzyloxy)-2-(3,4-bis(benzyloxy)phenyl)-3-hydroxy-4H-benzopyran-4-one 7 (500 mg; 0.754 mmol) in anhydrous DMF (4 ml). The reaction mixture was stirred at a temperature of 65°C for 12 hours. After completion, the mixture was partitioned between water (10 ml) and EtOAc (20 ml). The organic phase was washed with a saturated aqueous NaCl solution and the aqueous phase was extracted with EtOAc (20 ml). The combined organic phases were dried over anhydrous Na2SO4 and the solvent was removed under reduced pressure. The residue obtained in the form of a yellow oil was purified by flash column chromatography (SiO2; EtOAc / n-hexane 25%). The product 11b was isolated as a white oil (460 mg, 72%).

[0138]

[0139] 1H NMR (400MHz, DMSO-d6): 7.83 (bs, 1H), 7.63 (d, 1H, J = 4.0Hz), 7.46-7.18 (m, 21H), 6.95 (d, 1H, J = 2.0Hz), 6.76 (bt, 1 H, J = 8.0Hz), 6.70 (d, 1H, J = 2.0Hz), 5.25 (m, 8H), 4.12 (m, 2H), 3.60 (m, 2H), 3.32 (m, 2H), 3.03 (m, 2H), 1.35 (s, 9H).

[0140] 13C NMR(100MHz,DMSO-d6):173.2,163.6,159.0,156.5,152.5,151.2,148.6,140.3,138.1,137.8,137.7,137.0,129.5,129.4,129.3,129.2, 129.0,128.9,128.8,128.6,128.5,127.8,123.9,123.3,115.1,114. 6,109.8,98.7,95.1,78.6,71.6,71.4,71.0,70.9,70.3,70.0,29.1.

[0141] Synthesis example of compound 11c.

[0142] 2-[2-(Benzyloxy)ethoxy]ethyl bromide (390 mg; 1.508 mmol), anhydrous K2CO3 (250 mg; 1.810 mmol) and KI (36 mg; 0.226 mmol) were added successively to a solution of 5,7-bis(benzyloxy)-2-(3,4-bis(benzyloxy)phenyl)-3-hydroxy-4H-benzopyran-4-one 7 (0.5 g; 0.754 mmol) in anhydrous DCM (4 ml). The reaction mixture was stirred at a temperature of 65° C. for 12 hours. After completion, the reactant was partitioned between water (20 ml) and EtOAc (20 ml). The organic phase was washed with saturated aqueous NaCl solution and the aqueous phase was extracted with EtOAc (10 ml). The combined organic phases were dried over anhydrous Na2SO4 and the solvent was removed under reduced pressure. The residue obtained in the form of a light yellow solid was purified by flash column chromatography (SiO2; EtOAc / n-hexane 25%). The product 11a was isolated as a white oil (539 mg, 85%).

[0143]

[0144] 1 H NMR (400MHz, DMSO-d6): 7.63 (d, 1H, J = 7.6Hz), 7.50 (d, 4H, J = 7.6Hz), 7.47-7.28 (m, 22H), 7.20 (d, 1H, J=8.4Hz), 6.93 (d, 1H, J=1.2Hz), 6.72 (d, 1H, J=1.2Hz), 5.25 (m, 6H), 5.04 (m, 2H), 4.95 (m, 2H).

[0145] 13 C NMR (100MHz, DMSO-d6):172.9,163.2,159.6,158.6,158.4,152.6,150.5,148.0,139.6,137.5,136.7,128.8,128.5,128. 4,128.3,128.2,128.1,128.0,127.9,127.3,123.3,122.2,114.6,114.1,109.3,98.3,94.6,73.3,70.6,70.4,70.3,70.2.

[0146] Synthesis example of compound 12.

[0147] An aqueous solution of 2N NaOH (2 ml) was added to a solution of 11a (440 mg; 0.567 mmol) in ethanol (6 ml). The reaction mixture was stirred at room temperature for 12 hours. After completion, an aqueous solution of 2N HCl (-2.5 ml) was added to the reaction mixture to pH ~ 3, and then extracted with DCM (3×10 ml). The combined organic phases were dried over anhydrous Na2SO4 and the solvent was removed under reduced pressure; and the residue 12 in the form of a white solid was used in the subsequent reaction without further chromatographic purification (quantitative).

[0148]

[0149] Synthesis example of compound 16

[0150] A 4N aqueous solution of HCl (4 ml) was added to a solution of 11a (460 mg; 0.541 mmol) in THF (6 ml). The reaction mixture was stirred at 50° C. for 6 hours. After completion, the reaction mixture was dried under reduced pressure and the residue 16 in the form of a white solid was used in the subsequent reaction without further chromatographic purification (quantitative).

[0151]

[0152] 1 H NMR (400MHz, CD3OD): 7.66-7.30 (m, 22H), 7.05 (d, 1H, J = 8.4Hz), 6.57 (d, 1H, J = 2.0Hz), 6.51 (d, 1H, J = 2.0Hz), 5.27 (s ,2H),5.25(s,2H),5.17(s,2H),5.12(2H),3.83-3.74(m,2H,3.72-3.64(m,2H),3.60-3.53(m,2H),3.22-3.12(m,2H).

[0153] Example of a general procedure for the synthesis of product 14.

[0154] HOBt (32 mg, 0.24 mmol), EDC*HCl (46 mg; 0.24 mmol) and DIPEA (140 μl; 0.8 mmol) were added sequentially to a solution of 12 (158 mg; 0.2 mmol) in anhydrous DCM (6 ml). The reaction mixture was stirred at room temperature for 15-30 minutes and then added to a solution of the selected diamine 13 (0.08 mmol) in anhydrous DCM (6 ml) and a catalytic amount of DMAP (~1 mg, 0.008 mmol) was added. The reaction mixture was stirred at room temperature for 12 hours. After completion, the reaction mixture was diluted with DCM (10 ml) and washed sequentially with saturated aqueous NH4Cl solution (3×10 ml), saturated aqueous NaHCO3 solution (3×10 ml) and finally saturated aqueous NaCl solution (1×10 ml). The combined organic phases were dried over anhydrous Na2SO4 and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography.

[0155]

[0156] Compound 14 was isolated as a white oil (274 mg, 87.5%).

[0157] 1 H NMR (400MHz, CDCl3): 7.79 (bd, 2H, J = 7.6Hz), 7.65 (d, 2H, J = 2.0Hz), 7.58-7.50 (m, 6H), 7. 47-7.25(m,36H),6.83(d,2H,J=8.8Hz),6.31(dd,4H,J1=7.1Hz,J2=2.1Hz),5.29(s,4H), 5.14(s,4H),5.05(s,4H),4.87(d,2H,J=11.3Hz),4.77(d,2H,J=11.3Hz),4.23(m,2H),3. 95(m,2H),3.85(m,2H),2.53(m,4H),2.02(m,4H),1.94(m,2H),1.58(m,2H),1.36(m,4H).

[0158] 13C NMR(100MHz, CDCl3):173.7,172.7,162.6,159.5,158.4,152.5,150.9,148.3,139.9,137.0,136.6,136.4,135.5,128.6,128.5,128.2,127. 9,127.8,127.7,127.3,127.2,127.1,126.5,123.5,122.2,114.8,113 .4,109.6,97.9,93.5,71.5,70.6,70.5,70.1,49.0,33.1,28.4,27.0.

[0159]

[0160] Compound 14b was isolated as a white oil (312 mg, 93%).

[0161] 1 H NMR (400MHz, CDCl3): 7.75 (d, 2H, J = 2.0Hz), 7.68 (dd, 2H, J1 = 10.4Hz, J2 = 2.0Hz), 7.59 (d, 4H, J = 8.0Hz), 7. 48(t,8H,J=6.4Hz),7.45-7.30(m,28H),7.16(bt,2H,J=5.6Hz),7.03(d,2H,J=8.0Hz),6.52(d,2H,J=2.0Hz ),6.54(d,2H,J=2.0Hz),5.19(s,8H),5.15(s,4H),5.03(s,4H),3.88(t,4H,J=5.6Hz),3.53(m,4H),3.47( m, 4H), 3.41 (t, 4H, J = 6.4Hz), 3.27 (q, 4H, J = 12.4, 6.4Hz), 2.46 (t, 4H, J = 6.8Hz), 1.97 (m, 4H), 1.73 (m, 4H).

[0162] 13C NMR (100MHz, CDCl3):173.9,173.0,162.8,159.7,158.6,153.6,150.9,1 48.4,140.0,137.0,136.6,136.3,135.6,128.7,128.5,128.4,128.0,127 .9,127.7,127.5,127.3,127.2,126.7,123.6,122.4,115.1,113.8,109. 8,97.9,93.8,71.6,70.8,70.7,70.4,70.0,69.3,37.1,33.1,29.3,26.7.

[0163]

[0164] Compound 14c was isolated as a white oil (298 mg, 93.2%).

[0165] 1 H NMR (400MHz, CDCl3): 7.72 (d, 2H, J = 2.0Hz), 7.63 (dd, 2H, J1 = 8.4Hz, J2 = 2.0Hz), 7.55-7-20 (m, 44H), 7.01 (d, 2H, J = 8.8Hz), 6.56 (d, 2H, J = 2.4Hz), 6.45 (d,2H,J=2.0Hz),5.23(s,4H)5.22(s,4H),5.14(s,4H),5.12(s,4H),4.22( d, 4H, J = 5.6Hz), 3.82 (t, 4H, J = 5.2Hz), 2.52 (t, 4H, J = 6.4Hz), 1.96 (m, 4H).

[0166] 13 C NMR (100MHz, CDCl3):173.9,173.1,162.9,159.7,158.6,153.1,150.9,148.3,139.9,137.5,137.0,136.6,136.2,135.6,128.7,128.6,128.5,1 28.4,128.0,127.9,127.7,127.6,127.3,127.2,126.6,123.6,122.3,11 5.1,113.8,109.8,98.0,93.8,71.5,70.8,70.6,70.5,43.0,33.4,26.7.

[0167]

[0168] Compound 14d was isolated as a white oil (294 mg, 91.6%)

[0169] 1 H NMR (400MHz, CDCl3): 7.75 (bs, 2H), 7.67 (dd, 2H, J1 = 8.8Hz, J2 = 1.6Hz), 7.58 (d, 4H, J = 7 .6Hz),7.55-7.25(m,38H),7.04(d,2H,J=8.4Hz),6.91(bs,2H),6.54(d,2H,J=1.8Hz), 6.47(d,2H,J=1.6Hz),5.26(s,4H),5.25(s,4H),5.21(s,4H),5.11(s,4H),3.82(t,4H, J=5.2Hz),2.93(t,4H,J=6.0Hz),1.96(m,4H),1.51(m,4H),1.23(bs,2H),0.65(m,4H).

[0170] 13 C NMR (100MHz, CDCl3):174.2,173.9,162.9,159.7,158.7,153.2,151.0,1 40.0,136.9,136.6,136.3,135.6,128.7,128.6,128.5,128.4,128.0,127 .7,127.9,127.7,127.6,127.3,127.2,126.6,123.6,122.3,115.2,113.8 ,109.8,98.0,93.9,71.6,70.9,70.7,70.5,45.6,37.6,33.3,30.0,26.8.

[0171] Example of a general procedure for the synthesis of products 18, 21 and 24.

[0172] HOBt (38 mg, 0.28 mmol), EDC·HCl (54 mg; 0.28 mmol) and DIPEA (175 μl; 1.0 mmol) were added sequentially to a solution of the appropriate dicarboxylic acid linker 17 (0.10 mmol) or tricarboxylic acid linker 20 (0.062 mmol) or tetracarboxylic acid linker 23 (0.050 mmol) in anhydrous DCM (6 ml). The reaction mixture was stirred at room temperature for 15-30 minutes and then added to a solution of 16 (196 mg; 0.25 mmol) in anhydrous DCM (6 ml) and a catalytic amount of DMAP (~1 mg, 0.008 mmol) was added. The reaction mixture was stirred at room temperature for 12 hours. Upon completion, the reaction mixture was diluted with DCM (10 ml) and washed sequentially with saturated NH4Cl aqueous solution (3×10 ml), saturated NaHCO3 aqueous solution (3×10 ml) and finally saturated NaCl aqueous solution (1×10 ml). The combined organic phases were dried over anhydrous Na2SO4 and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography.

[0173]

[0174] Compound 18a was isolated as a white oil (298 mg, 93.4%).

[0175] 1 H NMR (400MHz, DMSO-d6): 7.83 (bs, 4H), 7.62 (d, 2H, J = 7.6Hz), 7.55-7.20 (m, 40H), 6.94 (s, 2H), 6.71 (s, 2H), 5.25 (bs, 16H), 4.13 (bs, 4H), 3.60 (bs, 4H), 3.44 (m, 4H), 3.14 (d, 4H, J = 5.2Hz), 2.17 (t, 4H, J = 7.6Hz), 2.07 (m, 4H), 1.68 (t, 2H, J = 7.6Hz).

[0176] 13 C NMR(100MHz,DMSO-d6):175.2,173.2,172.7,163.6,160.0,159.0,152.5,151.2,148.6,140.3,138.1,137.8,137.7,137.0,129.5,129.4,129 .3,129.2,129.0,128.9,128.6,128.5,127.9,123.9,123.3,115.1,11 4.6,109.8,98.0,95.1,71.6,71.4,71.0,70.9,70.4,70.0,35.3,19.5.

[0177]

[0178] Compound 18b was isolated as a white oil (264 mg, 77.4%).

[0179] 1 H NMR (400MHz, DMSO-d6): 8.54 (bt, 2H, J = 5.4Hz), 7.92 (d, 4H, J = 8.4Hz), 7.84 (m, 4H), 7.43 (d, 4H, J = 8.4Hz), 7.61 (d, 4H, J = 7.2Hz) 7.55 -7.15(m,38H),7,93(d,2H,J=1.6Hz),7.70(d,2H,J=2.0Hz),5.24(s,16H),4.15(bm,4H),3.67(bm,4H),3.52(bm,4H),3.42(bm,4H).

[0180] 13 C NMR(100MHz,DMSO-d6):173.4,166.4,163.8,160.1,157.9,152.3,151.2,148.4,142.2,140.2,138.3,137.8,137.7,137.0,134.5,129.5, 129.4,129.3,129.0,128.8,128.6,127.8,127.4,124.3,123.5,115. 2,114.8,110.4,98.9,95.7,71.5,71.4,71.2,71.0,70.9,70.6,70.0.

[0181]

[0182] Compound 18c was isolated as a white oil (236 mg, 68.5%).

[0183] 1H NMR (400MHz, CDCl3): 7.84 (d, 2H, J = 2.0Hz), 7.76 (d, 4H, J = 8.8Hz), 7.69 (dd, 2H, J1 =8.4Hz, J2=1.6Hz),7.53(d,4H,J=7.6Hz),7.50-7.20(m,44H),6.97(d,2H,J=8.8H z),6.78(d,4H,J=8.8Hz),6.51(d,2H,J=2.0Hz),6.41(d,2H,J=2Hz),5.25(s,4H), 5.21(s,4H),5.11(s,4H),5.07(s,4H),4.15(bm,4H),3.72(bm,4H),3.61(bm,8H).

[0184] 13 C NMR (100MHz, CDCl3):173.8,166.7,162.9,159.6,158.7,158.6,152.7,151.0 ,148.2,140.0,136.9,136.5,136.2,135.6,129.8,129.3,128.7,128.6,128. 5,128.4,128.0,127.9,127.7,127.6,127.4,127.2,126.6,123.6,122.5,118.3,115.7,113.6,109.7,98.0,93.8,71.7,71.2,70.8,70.6,70.5,70.2,69.5.

[0185]

[0186] Compound 18d was isolated as a white oil (242 mg, 66.4%).

[0187] 1H NMR (400MHz, CDCl3): 7.82 (d, 2H, J = 1.6Hz), 7.74 (d, 4H, J = 3.2Hz), 7.56 (d, 4H, J = 7.2H z),7.50-7.25(m,38H),7.01(bs,2H),6.98(d,2H,J=8.8Hz),6.75(d,J=8.8Hz),6.53( d,2H,J=2.0Hz),6.44(d,2H,J=2.0Hz),5.23(s,4H),5.18(s,8H),5.08(s,4H),.4.18( m,2H),3.85(t,4H,J=6.4Hz),3.71(m,4H),3.58(bm,8H),1.74(bt,4H),1.46(bt,4H).

[0188] 13 C NMR (100MHz, CDCl3):173.7,167.0,162.8,161.4,159.7,158.6,152.7,151.0,148 .2,148.2,140.0,137.0,136.6,136.2,135.6,128.9,128.7,128.5,128.4,128.0,1 27.9,127.7,127.6,127.4,1272,126.6,123.7,122.6,115.7,113.9,113.6,109.9,98.0,93.8,71.7,71.2,70.8,70.7,70.5,70.3,69.7,67.8,39.8,37.0,29.0,25.8.

[0189]

[0190] Compound 21a was isolated as a white oil (283 mg, 58.8%).

[0191] 1H NMR (400MHz, CDCl3-CD3OD 80:20):8.32(s,3H)7.96(bt,3H,J=5.2Hz),7.65(d,6H,J=8.8Hz),7.45-7.15(m, 60H); 6.91 (d, 3H, J = 8.8Hz), 6.46 (d, 3H, J = 1.6Hz), 6.38 (d, 3H, J = 1.6Hz), 5.10 (sm 6H),5.07(s,6H),5.06(s,6H),4.99(s,6H),4.38(bs,3H),3.96(bm,6H),3.54(bm,6H),3.44(m,6H),3.39(m,6H).

[0192] 13 C NMR (100 MHz, CDCl3-CD3OD 80:20):174.3,166.6,163.0,159.5,158.5,153.1,150.9,148.0,139.8,136.8,136.4,136.0,135.6,134.5,128.8,128.5,128.4,128.3,128.2,127.9,127.8,127.6,127.4,127.3,127.2,126.7,123.2,122.7,115.1,113.5,109.4,97.9,93.9,71.4,71.3,70.7,70.6,70.4,70.0,69.3,39.9.

[0193]

[0194] Compound 21b was isolated as a white oil (293 mg, 60.7%).

[0195] 1H NMR (400MHz, CDCl3-CD3OD 80:20):7.79(bs,3H),7.71(d,3H,J=8.4Hz),7.57(d,6H,J=7.6Hz),7.50–7.25(m,54H),6. 98(d,3H,J=8.8Hz),6.53(d,3H,J=1.8Hz),6.44(d,3H,J=1.8Hz),6.22(bt,3H),5.21(s,8H) ,5.20(s,16H),5.06(s,8H),4.13(bs,6H),3.60(bs,6H),3.34(t,6H,J=4.8Hz),3.23(t,6H, J=4.8Hz), 1.98(bs,3H), 1.88(t,3H,J=12Hz), 1.78(d,3H,J=12Hz), 1.42(q,3H,J=12.8Hz).

[0196] 13 C NMR (100 MHz, CDCl3-CD3OD 80:20):174.3,173.6,162.8,159.7,158.6,152.7,150.9,148.2,140.0,137.0,136.6,136.3,135.6,128.7,128.5,128.4,128.0,127.9,127.7,127.6,127.4,127.2,126.6,123.6,122.7,115.6,113.7,109.8,98.0,93.9,71.6,71.2,70.8,70.7,70.4,70.1,69.7,43.5,39.1,31.5.

[0197]

[0198] Compound 24a was isolated as a white oil (385 mg, 59.1%).

[0199] 1 H NMR (400MHz, CDCl3-CD3OD 80:20):8.17(s,2H),8.09(s,4H),7.69(d,4H,J=1.6Hz),7.65(dd,4H,J1=10.4Hz,J2=2.0Hz),7.4 5-7.22 (m, 80H), 7.18 (m, 4H), 6.92 (d, 4H, J = 8.8Hz), 6.46 (d, 4H, J = 1.6Hz), 6.36 (d, 4H, J = 1.6Hz).

[0200] 13C NMR (100MHz, CDCl3-CD3OD 80:20):171.5,164.1,158.8,154.0,151.7,148.7,143.8,139.1,136.8,136 .4,135.8,135.0,134.3,128.6,128.5,128.4,128.3,128.0,127.9,127.8,12 7.7,127.6,127.4,127.2,126.9,126.6,124.0,122.8,114.7,111.9,105.2,95.4,93.3,72.8,71.4,71.3,71.0,70.7,70.6,70.5,70.4,70.2,69.7,37.6.

[0201] Examples of a general method for debenzylation of 3-O-alkylated quercetin: 2-(3,4-dihydroxyphenyl)-3-ethoxy-5,7-dihydroxy-4H-benzopyran-4-one 1; 2-(3,4-dihydroxyphenyl)-3-hexyloxy-5,7-dihydroxy-4H-benzopyran-4-one 2; 2-(3,4-dihydroxyphenyl)-3-hexadecyloxy-5,7-dihydroxy-4H-benzopyran-4-one 3.

[0202] The target product solution dissolved in EtOH / THF (1:2% v / v) mixture was transferred to a hydrogenation flask. Catalyst Pd / C (10%) was added to the mixture. The reaction mixture was subjected to vacuum-nitrogen cycles three times to remove air from the system; after the last vacuuming, hydrogen was charged into the flask to a pressure of up to 1.2 bar. Finally, the reaction mixture was stirred at room temperature until the desired product was completely debenzylated (about 18 hours). After completion, the catalyst was removed from the reaction mixture by filtration, and the solvent was removed under reduced pressure. The final residue corresponding to the desired product did not require further final purification.

[0203]

[0204] 2-(3,4-dihydroxyphenyl)-3-ethoxy-5,7-dihydroxy-4H-benzopyran-4-one 1:

[0205]

[0206] 1H NMR (400MHz, CDCl3): 7.62 (d, 1H, J = 2.1Hz), 7.54 (dd, 1H, J1 = 8.5Hz, J2 = 2.1Hz), 6.89 (d, 1H, J = 8. 5Hz), 6.37 (d, 1H, J = 2.1Hz), 6.18 (d, 1H, J = 2.1Hz), 3.89 (q, 2H, J = 7.1Hz), 1.32 (t, 1H, J = 7.1Hz);

[0207] 13 C NMR(100MHz, CDCl3):180.29,166.85,163.22,158.66,158.39,150.07,146.56,138.53,123.34,122.55,1 22.26,116.71,116.46,116.42,116.16,105.78,100.15,99.86,95.02,94.73,69.77,69.49,15.82,15.55.

[0208] 2-(3,4-dihydroxyphenyl)-3-hexyloxy-5,7-dihydroxy-4H-benzopyran-4-one 2:

[0209]

[0210] 1 H NMR (400MHz, CDCl3): 7.55 (s, 1H); 7.47 (d, 1H, J = 8.2Hz), 6.86 (d, 1H, J = 8.2Hz), 6.35 (s, 1H), 6 .17(s,1H),3.88(t,1H,J=6.0Hz),1.75-1.60(m,2H),1.50-1.10(m,6H),0.87(t,3H,J=6.0Hz);

[0211] 13 C NMR (100MHz, CDCl3):180.27,165.90,163.22,158.54,149.87,146.44,138.79,123.31,1 22.68,116.87,116.31,106.06,99.84,94.80,74.18,32.86,31.11,26.89,23.78,14.52.

[0212] 2-(3,4-dihydroxyphenyl)-3-hexadecyloxy-5,7-dihydroxy-4H-benzopyran-4-one 3:

[0213]

[0214] 1 H NMR (400MHz, CDCl3): 7.49 (d, 1H, J = 2.1Hz), 7.45 (dd, 1H, J1 = 8.5Hz, J2 = 2.1Hz), 6.81 (d, 1H, J = 8.5Hz), 6.27 (d, 1H, J =2.1Hz),6.15(d,1H,J=2.1Hz),3.78(t,3H,J=7.1Hz),1.68-1.55(m,2H),1.26-1.09(m,30H),0,76(t,3H,J=7.1Hz);

[0215] 13 C NMR(100MHz, CDCl3):178.72,163.64,161.26,156.80,156.58,147.52,144.25,137.42,122.11,121.52,115.18,114.82,104.91 ,98.60,93.75,49.16,48.95,48.74,48.52,48.30,31.67,29.68,29.44,29.40,29.36,29.32,29.17,29.10,25.60,22.41,13.75.

[0216] Synthesis Example of 2-(3,4-bis(palmitoyloxy)phenyl)-4-oxo-4H-benzopyran-3,5,7-triacetate: 10a.

[0217] Acetyl chloride (0.850 ml; 0.012 mol), DIPEA (3.09 ml; 0.018 mol) and DMAP (0.36 g; 0.003 mol) were added sequentially to a solution of 3,3',4',5,7-pentahydroxyflavone 9 (0.56 g; 0.0016 mol) in anhydrous DCM (50 ml). The reaction mixture was stirred at room temperature. After completion (after 24 h), a white precipitate was formed. The reaction mixture was partitioned between H2O and DCM, and the combined organic phases were dried over anhydrous Na2SO4, filtered, and the solvent was removed under reduced pressure. The solid residue was recrystallized from MTBE (tert-butyl methyl ether) to give a yellow solid product corresponding to 10a (74.7%).

[0218] Synthesis Example of 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4-oxo-4H-benzopyran-3-yl acetate: 4

[0219] Will and butanol (1 ml; 0.01 mol) were added sequentially to a suspension of 2-(3,4-bis(palmitoyloxy)phenyl)-4-oxo-4H-benzopyran-3,5,7-triacetate 10a (0.512 g; 0.001 mol) in MTBE. The reactants were placed in an orbital oscillator (300 rpm / min) at a temperature of 45°C until complete conversion to the desired product. The progress of the reaction was monitored by TLC (DCM and DCM / MeOH=9 / 1). After completion (about 9 days), the solution was filtered, the solvent was removed under reduced pressure, and the residue was purified by flash column chromatography (silica diol: DCM / MeOH=9 / 1). The product 4 was thus isolated as a yellow solid (260 mg, 76.3%).

[0220]

[0221] 1 H NMR (400MHz, CDCl3): 7.45 (d, 1H, J = 2.1Hz), 7.37 (dd, 1H, J1 = 8.5Hz, J2 = 2.1Hz), 6.99 (d, 1H, J = 8.5Hz), 6.5 3(d,1H,J=2.1Hz),6.30(d,1H,J=2.1Hz),2.63(t,2H,J=7.4Hz),1.79-1.62(m,2H),0.87(t,3H,J=7.4Hz); 13 C NMR(100MHz, CDCl3):176.58,171.22,165.31,162.79,158.12,157.32,149.55, 146.05,131.46,122.00,121.95,116.36,115.95,105.12,99.86,23.31,14.34.

[0222] Synthesis Example of 2-(3,4-bis(palmitoyloxy)phenyl)-4-oxo-4H-benzopyran-3,5,7-triyl tripalmitate: 10b.

[0223] Palmitoyl chloride (3.51 ml; 0.012 mol), DIPEA (3.09 ml; 0.018 mol) and DMAP (0.36 g; 0.003 mol) were added sequentially to a solution of 3,3',4',5,7-pentahydroxyflavone 9 (0.56 g; 0.0016 mol) in anhydrous DCM (50 ml). The reaction mixture was stirred at room temperature. After completion (after 24 hours), a white precipitate was formed.

[0224] The reaction mixture was partitioned between H2O and DCM, and the combined organic phases were dried over anhydrous Na2SO4, filtered, and the solvent was removed under reduced pressure. The solid residue was recrystallized from MTBE, thereby obtaining the product corresponding to 10b as a yellow solid (64.7%).

[0225] Synthesis Example of 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4-oxo-4H-benzopyran-3-yl palmitate: 5

[0226] Will (2g) and butanol (1ml; 0.01mol) were added successively to a suspension of 2-(3,4-bis(palmitoyloxy)phenyl)-4-oxo-4H-benzopyran-3,5,7-triyltripalmitate 10b (1.3g; 0.001mol) in MTBE. The reactants were placed in an orbital oscillator (300rpm / min) at a temperature of 45°C until complete conversion to the desired product. The progress of the reaction was monitored by TLC (DCM and DCM / MeOH=9 / 1). After completion (about 9 days), the solution was filtered, the solvent was removed under reduced pressure, and the residue was purified by flash column chromatography (silica diol: DCM / MeOH=9 / 1). Thus, the yellow solid product 5 (250mg, 46.3%) was isolated.

[0227]

[0228] 1 H NMR (400MHz, CDCl3): 7.45 (d, 1H, J = 2.1Hz), 7.37 (dd, 1H, J1 = 8.5Hz, J2 = 2.1Hz), 6.99 (d, 1H, J = 8.5Hz), 6.53 (d, 1H, J = 2.1Hz ), 6.30 (d, 1H, J = 2.1Hz), 2.63 (t, 2H, J = 7.4Hz), 1.79-1.62 (m, 2H), 1.47-1.36 (m, 2H), 1.28 (s, 22H), 0.87 (t, 3H, J = 7.4Hz); 13 CNMR(100MHz, CDCl3):176.58,171.22,165.31,162.79,158.12,157.32,149.55,146.05,131.46,122.00,121.95,116.36,115.95,10 5.12,99.81,99.76,94.89,34.28,32.63,30.42,30.29,30.22,30.10,30.03,29.90,29.84,29.65,29.46,29.26,25.55,23.31,14.34.

[0229] Example of a general debenzylation procedure.

[0230] A solution of the desired product (0.01 mmol) in a mixture of EtOH / THF (1:2% v / v) was transferred to a hydrogenation flask. Catalyst Pd / C (10%) was added to the mixture. The reaction mixture was subjected to three vacuum-nitrogen cycles to remove air from the system; after the last vacuum, hydrogen was charged into the flask to a pressure of up to 1.2 bar. Finally, the reaction mixture was stirred at room temperature until the desired product was completely debenzylated (about 12 hours). After completion, the catalyst was removed from the reaction mixture by filtration, and the solvent was removed under reduced pressure. The residue was washed with a mixture of n-hexane / Et2O 70:30 (3×10 ml). Unless otherwise stated, the final residue corresponding to the desired product did not require final chromatographic purification.

[0231]

[0232] The product 15a was isolated as a yellow amorphous solid (65 mg, 76%).

[0233] 1 H NMR(400MHz,DMSO-d6):12.70(s,2H),10.85(s,2H),9.73(bs,2H),9.39(bs ,2H),7.55(bs,2H),7.45(d,2H,J=8.4Hz),7.39(d,2H,J=8.0Hz),6.89(d,2 H,J=8.4Hz),6.40(s,2H),6.18(s,2H),3.89(m,6H),2.22(q,4H,J=6.8Hz), 1.87(t,4H,J=6.8Hz),1.55(m,4H),1.43(m,4H),1.35(m,2H),1.23(m,2H).

[0234] 13 C NMR (100MHz, CD3OD):178.5,176.3,174.6,171.6,164.0,163.6,161.2,158.5,156.8,148.1,145.4,144.5,137.0,136.8,125.7,1 24.6,121.6,121.4,121.1,116.5,115.4,115.2,108.9,107.0,104.8,98.8,96.2,93.9,67.4,33.9,30.0,27.8,25.7,19.6,17.6.

[0235]

[0236] 15b

[0237] The product 15b was isolated as a yellow amorphous solid (70 mg, 72.8%).

[0238] 1 H NMR (400MHz, CD3OD): 7.57 (s, 2H, J = 2.4Hz), 7.45 (dd, 2H, J = 8.4, 2.0Hz), 6.88 (d, 2H, J = 8.4Hz), 6.31 (dd, 2H, J = 2.0Hz), 6.14 (d, 2H, J = 2.0Hz), 3.89 (t, 4 H,J=6.0Hz),3.58(m,4H),3.52(m,4H),3.48(t,4H,J=6.4Hz),3.27(t,4H,J= 6.8Hz), 2.40 (t, 4H, J = 7.2Hz), 2.01 (t, 4H, J = 6.4Hz), 1.74 (t, 4H, J = 6.4Hz).

[0239] 13 C NMR(100MHz,CD3OD):178.5,176.3,164.0,163.6,161.2,158.5,156.8,148.1,145.4,137.0,124 .7,121.6,115.0,104.8,98.4,93.3,71.3,68.5,57.0,36.6,33.9,32.2,28.8,27.4,26.0,16.9.

[0240]

[0241] The product 15c was isolated as a yellow amorphous solid (82 mg, 93.5%).

[0242] 1 H NMR (400MHz, CD3OD): 7.52 (d, 2H, J = 2.0Hz), 7.41 (dd, 4H, J = 8.4, 2.0Hz), 7.09 (s, 4H); 6.83 (d, 2H, J = 8.8Hz), 6.26 (d, 2H, J=1.6Hz), 6.09 (d, 2H, J=2.0Hz), 4.25 (s, 4H), 3.83 (t, 4H, J=6.0Hz), 2.42 (t, 4H, J=6.8Hz), 1.99 (t, 4H, J=6.4Hz).

[0243] 13C NMR(100MHz,CD3OD):178.3,176.4,164.17,161.4,156.7,156.6,148.3,144.8,137.2,13 6.8,127.4,121.5,121.0,115.16,114.9,104.4,98.3,93.3,71.2,65.4,42.6,32.4,26.0.

[0244]

[0245] The product 15d was isolated as a yellow amorphous solid (74 mg, 83.8%).

[0246] 1 H NMR (400MHz, CD3OD): 7.57 (bs, 1H), 7.47 (d, 2H, J = 8.0Hz), 6.87 (d, 2H, J = 8.0Hz), 6.31 (bs, 2H), 6.13 (m,2H),3.89(m,4H),2.99(m,4H),2.39(m,4H),2.00(m,4H),1.70(m,4H),1.36(m,2H),0.86(m,4H).

[0247] 13 C NMR(100MHz,CD3OD):178.5,174.3,164.3,161.5,156.9,156.7,148.4,144.8,13 6.9,121.6,121.0,115.2,114.9,104.4,98.3,93.3,71.3,37.7,32.3,29.9,26.0.

[0248]

[0249] The product 19a was isolated as a yellow amorphous solid (85 mg, 97.2%).

[0250] 1 H NMR (400MHz, DMSO-d6): 7.82 (s, 2H), 7.56 (m, 2H), 6.88 (d, 2H, J = 8.4Hz), 6.40 (s, 2H), 6.18 (s, 2H), 4.26 (m, 4H ), 3.64 (m, 4H), 3.63 (t, 4H, J = 5.6Hz), 3.18 (m, 4H), 2.17 (t, 4H, J = 6.8Hz), 2.09 (t, 4H, J = 7.2Hz), 1.69 (m, 4H).

[0251] 13C NMR(100MHz,DMSO-d6):178.8,175.4,172.8,165.5,162.2,157.3,156.6,149.9,146.2, 137.5,122.0,121.7,116.6,104.9,99.6,94.5,72.0,70.3,70.0,35.4,34.3,31.4,21.8.

[0252]

[0253] The product 19b was isolated as a yellow amorphous solid (95 mg, 96.4%).

[0254] 1 H NMR (400MHz, DMSO-d6): 12.70(s,2H),10.92(s,2H),9.81(s,2H),9.38(s,2H),8.57(bt,2H),7.95(d,4H,J=8.0Hz),7.79(d,4H,J=8.0Hz ),7.57(m,2H),6.92(d,2H,J=8.4Hz),6.43(d,2H,J=1.6Hz),6.20(d,2H,J=1.6Hz),4.15(m,4H),3.71(m,4H),3.53(m,4H),3.43(m,4H).

[0255] 13 C NMR(100MHz,DMSO-d6):178.8,166.9,165.1,162.2,157.2,156.7,149.6,146.6,146.1,142.6,140.1, 137.6,134.6,129.2,128.9,127.60,125.9,122.1,121.9,116.6,105.1,99.5,72.0,70.3,69.8,69.2.

[0256]

[0257] The product 19c was isolated as a yellow amorphous solid (87 mg, 86.9%).

[0258] 1H NMR (400MHz, CD3OD): 7.74 (d, 4H, J = 8.0Hz), 7.63 (bs, 2H), 7.48 (d, 2H, J = 8.4Hz), 6.91 (d, 4H, J = 8.0Hz) ,6.82(d,2H,J=8.4Hz),6.26(s,2H),6.08(s,2H),4.05(m,4H),3.72(m,4H),3.60(m,4H),3.56(m,4H).

[0259] 13 C NMR(100MHz,CD3OD):178.3,168.2,164.2,161.5,159.1,156.8,156.5,148.4,144.7,136.8, 129.5,129.1,121.5,121.2,118.2,115.5,114.8,104.4,98.3,93.3,71.3,69.7,69.1,39.7.

[0260]

[0261] The product 19d was isolated as a yellow amorphous solid (93 mg, 84.5%).

[0262] 1 H NMR (400MHz, CD3OD): 7.68 (m, 6H), 7.53 (dd, 2H, J = 8.4, 2.0Hz), 6.87 (d, 2H, J = 8.4Hz), 6.81 (d, 4H, J = 8.8Hz), 6.33 (d, 2H, J = 1.6Hz), 6 .17(d,2H,J=1,6Hz),4.08(m,4H),3.92(t,4H,J=6.0Hz),3.74(m,4H),3.63(m,4H),3.57(t,4H,J=4.4Hz),1.75(m,4H),1.48(m,4H).

[0263] 13 C NMR (100MHz, CD3OD):178.4,168.7,164.2,161.8,161.5,156.9,156.7,148.4,144.7,136.9,128.7,1 26.0,121.6,121.3,115.7,114.9,113.8,104.6,98.5,93.5,71.4,69.8,69.3,67.6,39.6,28.7,25.4.

[0264]

[0265] The product 22a was isolated as a yellow amorphous solid (132 mg, 99.7%).

[0266] 1 H NMR (400MHz, CD3OD): 8.32 (s, 3H), 7.60 (d, 3H, J = 2.0Hz), 7.47 (dd, 3H, J = 8.4, 2.0Hz), 6.82 (d, 3H, J = 8.4Hz), 6 .27(d,3H,J=2.4Hz),6.09(d,3H,J=2.0Hz),4.02(m,6H),3.70(m,6H),3.65(m,6H),3.58(m,6H),3.53(m,6H).

[0267] 13 C NMR (100MHz, CD3OD):181.8,178.1,172.6,167.3,166.0,161.4,159.4,156.9,156.4,148.6,144 .8,136.6,134.9,128.5,121.4,121.2,115.4,114.9,103.9,98.9,93.7,71.3,69.7,69.0,39.8.

[0268]

[0269] The product 22b was isolated as a yellow amorphous solid (126 mg, 94.7%).

[0270] 1 H NMR (400MHz, CD3OD): 7.61 (bs, 3H), 7.49 (d, 3H, J = 6.8Hz), 6.84 (bs, 3H), 6.25 (m, 3H), 6.12 (m, 3H ),3.98(bs,6H),3.65(bs,6H),3.46(m,6H),3.35(m,3H),2.32(bs,3H),1.88(m,3H),1.59(m,3H).

[0271] 13 C NMR(100MHz,CD3OD):178.3,176.0,164.2,161.4,156.7,156.5,148.3,144.7,136.8 ,121.5,121.2,115.5,114.8,104.3,98.3,93.3,71.3,69.6,68.9,43.2,39.0,31.3.

[0272]

[0273] The product 25a was isolated as a yellow amorphous solid (142 mg, 78.2%).

[0274] 1 H NMR (400MHz, CD3OD): 8.09 (s, 2H), 7.94 (d, 4H, J = 1.2Hz), 7.53 (d, 4H, J = 1.2Hz), 7.42 (dd, 4H, J = 8.4, 2.4Hz), 6.75 (dd, 4 H,J=8.4,1.6Hz),6.22(s,4H),6.04(s,4H),4.06(bs,8H),3.74(m,8H),3.61(bt,8H,J=4.8Hz),5.52(bt,8H,J=4.8Hz).

[0275] 13 C NMR(100MHz,CD3OD):178.2,167.6,164.2,161.4,156.9,156.2,148.2,144.6,139.5,136.8, 134.9,128.2,125.5,121.5,121.2,115.4,114.8,104.3,98.2,93.3,71.1,69.7,68.8,39.8.

[0276]

[0277] The product 26 was isolated as a yellow amorphous solid (74 mg, 95.0%).

[0278] 1 H NMR(400MHz,DMSO-d6):12.47(bs,1H),9.36(bs,1H),7.67(bs,1H),7.53(d ,1H,J=8.4Hz),6.87(m,1H),6.41(bs,1H),6.18(bs,1H),3.45-3.20(m,8H).

[0279] 13 C NMR (100MHz, DMSO-d6): 175.9, 164.0, 160.8, 156.2, 147.8, 146.9, 145.1, 135.8, 125.0, 122.0, 120.1, 115.7, 115.1, 103.1, 98.2, 93.4.

[0280]

[0281] The product 27 was isolated as a yellow amorphous solid (74 mg, 95.0%).

[0282] 1 H NMR (400MHz, CD3OD): 7,67 (d, 1H, J = 2.0Hz), 7.52 (dd, 1H, J1 = 8.4Hz, J2 = 2.0Hz), 6.39 (d, 1H, J = 1.6Hz ),6.19(d,1H,J=2.0Hz),4.10-4.01(m,2H),3.83-3.75(2H,m),3.74-3.67(2H,m),3.19-3.10(2H,m); 13 C NMR(100MHz,CD3OD):179.9,166.0,163.0,158.5,158.4,150.0,146.3,138 .3,122.8,122.5,117.0,116.4,105.8,99.9,94.8,72.9,71.2,67.8,40.6.

[0283] Synthesis example of compound 29 (ag)

[0284] The method was carried out in the dark. EDC·HCl (1.5 eq.) was added to a solution of acid 28 (ag) (about 0.025 M) in anhydrous DMF under argon atmosphere and magnetic stirring, followed by HOBt (1.2 eq.) after 30 minutes. After another 30 minutes, the solution obtained as above was added dropwise to a solution of compound 27 (1.1 eq., about 0.029 M) and DIPEA (6-8 eq.) in anhydrous DMF by syringe over 5 minutes, with stirring, maintained at 0°C and under argon atmosphere. The reactant thus obtained was brought to room temperature and kept under stirring and argon atmosphere for 24 hours. After 24 hours, the reaction mixture was diluted with EtOAc (5 times the volume of the reactant) and extracted with water (twice) and saturated NaCl solution (once). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated to dryness in vacuo to afford the crude product, which was purified by flash column chromatography (SiO2; DCM / MeOH = 2-4%) to afford the final compound, ie, 29(ag).

[0285] Examples of synthesis conditions for individual compounds:

[0286] -29a: 27 (15 mg; 0.028 mmol), DIPEA (40 μl; 0.23 mmol); anhydrous DMF (1 ml); 28a (6.5 mg; 0.025 mmol); EDC·HCl (7.5 mg; 0.038 mmol); HOBt (3.8 mg; 0.028 mmol); anhydrous DMF (1.5 ml). Compound 29a (5 mg, 30%) was obtained as a yellow solid.

[0287]

[0288] 1 H NMR (400MHz, CD3OD): 7.68 (d, 1H, J = 2.0Hz), 7.52 (dd, 1H, J1 = 8.4Hz, J2 = 2.0Hz ),6.86(d,1H,J=8.4Hz),6.35(d,1H,J=2.0Hz),6.21(d,1H,J=2.0Hz),4.06-4, 00(m,2H),3.72-3.66(m,2H),3.49(t,2H,J=4.8Hz),3.36(t,2H,J=4.8Hz),2.1 2(t,2H,J=8Hz),1.58-1.46(m,2H),1.30-1.10(m,24H),0.81(t,3H,J=7.2Hz);

[0289] 13 C NMR (100MHz, CD3OD):178.8,175.2,164.3,161.6,157.2,157.0,148.3,144.8,137.3,122.0,121.7,115.9,11 5.3,105.1,99.1,94.2,71.7,70.1,69.8,39.4,32.0,29.8,29.76,29.6,29.5,29.47,29.41,26.0,22.8,14.1.

[0290] -29b: 27 (15 mg; 0.028 mmol), DIPEA (40 μl; 0.23 mmol); anhydrous DMF (1 ml); 28b (8.7 mg; 0.025 mmol); EDC·HCl (7.5 mg; 0.038 mmol); HOBt (3.8 mg; 0.028 mmol); anhydrous DMF (1.5 ml). Compound 29b (4.5 mg, 25%) was obtained as a yellow solid.

[0291]

[0292] 1H NMR (400MHz, CD3OD): 7.70 (d, 1H, J = 2.0Hz), 7.54 (dd, 1H, J1 = 8.4Hz, J2 = 1.6Hz) ,6.88(d,1H,J=8.4Hz),6.37(d,1H,J=2.4Hz),6.23(d,1H,J=2.4Hz),4.08-4.0 2(m,2H),3.75-3.68(m,2H),3.52(t,2H,J=5.2Hz),3.38(t,2H,J=5.2Hz),2.15 (t,2H,J=7.6Hz),1.62-1.48(m,2H),1.31-1.11(m,36H),0.84(t,3H,J=6.8Hz);

[0293] 13 C NMR (100MHz, CD3OD):179.0,175.5,164.6,161.9,157.4,157.3,148.6,145.1,137.5,122.2,121.9,116.2,115.5 ,105.3,99.3,94.3,71.9,70.3,70.0,39.6,36.7,32.2,30.9,30.0,29.8,29.69,29.65,29.60,26.2,23.0,14.2.

[0294] -29c: 27 (21 mg; 0.042 mmol), DIPEA (44 μl; 0.25 mmol); anhydrous DMF (1 ml); 28c (10.6 mg; 0.038 mmol); EDC·HCl (11 mg; 0.057 mmol); HOBt (5.6 mg; 0.042 mmol); anhydrous DMF (1.5 ml). Compound 29c (2 mg, 8%) was obtained as a yellow solid.

[0295]

[0296] 1H NMR (400MHz, CD3OD): 7.68 (d, 1H, J = 1.6Hz), 7.53 (dd, 1H, J1 = 8.4Hz, J2 = 1.6Hz), 6.8 6(d,1H,J=8,4Hz),6.35(d,1H,J=2.0Hz),6.21(d,1H,J=2.0Hz),5.29-5.22(m,2H),3 .72-3.65(m,2H),3.49(t,2H,J=5.2Hz),3.36(t,2H,J=5.2Hz),2.12(t,2H,J=7.6Hz ),1.98-1.85(m,4H),1.70-1.40(m,2H),11.35-1.10(m,20H),0,81(t,3H,J=7.6Hz);

[0297] 13 C NMR (100MHz, CD3OD):178.8,175.1,164.3,161.6,157.2,157.0,148.3,144.8,137.3,130.0,129.9,122.0,121.7,115 .9,115.2,105.1,99.1,94.2,71.7,70.1,69.8,39.4,36.6,32.0,29.9,29.8,29.6,29.4,29.3,27.3,26.0,22.8,14.1.

[0298] -29d: 27 (15 mg; 0.028 mmol), DIPEA (40 μl; 0.23 mmol); anhydrous DMF (1 ml); 28d (7.1 mg; 0.025 mmol); EDC·HCl (7.5 mg; 0.038 mmol); HOBt (3.8 mg; 0.028 mmol); anhydrous DMF (1.5 ml). Compound 29d (5 mg, 30%) was obtained as a yellow solid.

[0299]

[0300] 1H NMR (400MHz, CD3OD): 7.69 (d, 1H, 2.4Hz), 7.54 (dd, 1H, J1 = 8.4Hz J2=2.0Hz),6.88(d,1H,J=8.4Hz),6.37(d,1H,J=2.0Hz),6.22(d,1H,J=2.0Hz ),5.37-5.20(m,4H),4.08-4.01(m,2H),3.73-3.67(m,4H),3.51(t,2H,J=5.2 Hz), 3.38 (t, 2H, J = 5.2Hz), 2.70 (t, 2H, J = 6.4Hz), 21.47 (t, 2H, J = 7.6Hz), 2.0 4-1.92(m,4H),1.61-1.50(m,2H),1.35-1.13(m,14H),0.84(t,3H,J=7.2Hz);

[0301] 13 C NMR(100MHz,CD3OD):178.9,175.3,164.5,161.8,157.3,157.1,148.5,145.0,137.4,130.4,130.3,128.21,128.15,122.1,121.8 ,116.0,115.4,105.2,99.2,94.3,71.9,70.2,69.9,39.5,36.6,31.7,29.9,29.8,29.6,29.5,29.4,27.4,26.1,25.8,22.8,14.1.

[0302] -29e: 27 (14 mg; 0.026 mmol), DIPEA (27 μl; 0.16 mmol); anhydrous DMF (1 ml); 28e (7.2 mg; 0.024 mmol); EDC·HCl (6.8 mg; 0.036 mmol); HOBt (3.5 mg; 0.026 mmol); anhydrous DMF (1.5 ml). Compound 29e (5 mg, 31%) was obtained as a yellow solid.

[0303]

[0304] 1H NMR (400MHz, CD3OD): 7.69 (d, 1H, J = 2.0Hz), 7.54 (dd, 1H, J1 = 8.4Hz, J2 = 2.4Hz), 6.88 (d, 1H, J = 8.4Hz), 6.37 (d, 1H, J = 1.6Hz), 6.22 (d, 1H J=1.6Hz),5.38-5.21(m,8H),4.08-4.00(m,2H),3.74-3.66(m,2H),3.51(t,2H,J=5.2Hz),3.38(t,2H,J=5.2Hz),2.8 2-2.68(m,6H),2.17(t,2H,J=8,0Hz)2.1-1.94(m,4H),1.70-1.58(m,2H),1.35-1.15(m,6H),0.831(t,3H,J=6.8Hz);

[0305] 13 C NMR (100MHz, CDCl3):178.7,175.5,163.1,161.93,156.9,156.7,148.2,144.2,137.4,130.7,129.1,128.9,128.8,128.4,128.2,128.0, 127.6,122.3,121.6,116.1,115.1,105.6,99.4,94.3,71.6,70.1,69 .7,39.6,36.2,31.6,29.8,29.4,27.3,26.7,25.8,25.7,22.7,14.2.

[0306] -29f: 27 (25 mg; 0.05 mmol), DIPEA (63 μl; 0.36 mmol); anhydrous DMF (1.5 ml), 28f (15 mg; 0.046 mmol), EDC·HCl (13 mg; 0.069 mmol), HOBt (8 mg; 0.06 mol), anhydrous DMF (2.5 ml). Compound 29f (14 mg, 44%) was obtained as a yellow solid.

[0307]

[0308] 1H NMR(400MHz, CD3OD:7.69(d,1H,J=2.0),7.54(dd,1H,J1=8.8Hz,J2=2.4Hz),6.88(d,1H ,J=8.8Hz),6.37(d,1H,J=2.0Hz),6.22(d,1H,J=2.0),5.38-5.23(m,12H),4.08-4.01( m,2H),3.74-3.66(m,2H),3.51(t,2H,J=5.2Hz),3.38(t,2H,J=5.2Hz),2.85-2.70(m,1 0H),2.39-2.30(m,2H),2.22(t,2H,J=6.8Hz),2.08-1.97(m,2H),0.92(t,3H,J=7.2Hz);

[0309] 13 C NMR (100MHz, CD3OD):178.9,174.4,164.5,161.8,157.3,157.2,148.5,145.0,137.4,132.2,129.4,128.8,128.5,128.43,128.37,128. 33,128.13,127.3,122.2,121.8,116.1,115.4,105.3,99.2,94.3,71.9,70.2,69.9,39.6,36.3,25.83,25.81,25.74,23.8,20.8,14.3.

[0310] -29g: 27 (32 mg; 0.063 mmol), DIPEA (100 μl; 0.57 mmol), anhydrous DMF (2 ml), 28g (15.8 mg; 0.069 mmol), EDC·HCl (19.8 mg; 0.10 mmol); and HOBt (11.2 mg; 0.083 mmol), anhydrous DMF (3 ml). Compound 29g (9 mg, 22%) was obtained as a yellow solid.

[0311]

[0312] 1H NMR(400MHz, CD3OD):7.82-7.77(m,2H),7.68(d,1H,J=2.4Hz),7.59-7.47(m,5H),6,99-6.93(m,2H),6.89(d,1H,J=8.4Hz),6.34 (d,1H,J=1.6Hz),6.16(d,1H,J=1.6Hz),4.15-4.09(m,2H),3.81(s,3H),3.79-3.73(m,2H),3.66-3.61(m,2H),3.61-3.55(m,2H);

[0313] 13 C NMR (100MHz, CD3OD):179.9,170.3,165.8,163.0,161.2,158.3,158.0,149.9,146.3,145.2,138.3,133.52,13 3.48,129.1,128.8,127.3,123.0,122.6,117.0,116.3,115.3,105.8,99.8,94.7,72.8,71.3,70.6,55.7,41.1.

[0314] Examples of important reactions and syntheses, which are also relevant for obtaining the heterotrimer Q2E 36

[0315] Synthesis Example of Intermediate 31

[0316] Under magnetic stirring and argon atmosphere, EDC·HCl (379 mg; 1.98 mmol) and then, about 30 minutes later, HOBt (214 mg; 1.58 mmol) were added to a solution of palmitic acid (372 mg; 1.45 mmol) in anhydrous DMF (2 ml). The reaction mixture thus obtained was stirred for about thirty minutes and then added dropwise via a syringe over 5 minutes to a solution of dopamine·HCl (30; 250 mg; 1.32 mmol) and DIPEA (1.38 ml; 7.9 mmol) in anhydrous DMF (10 ml) maintained at 0° C. (in an ice bath).

[0317] After the addition was complete, the reaction mixture was allowed to warm to room temperature slowly and then stirred for 24 hours under argon atmosphere, then diluted with AcOEt (30 ml), washed successively with H2O (2×30 ml) and saturated NaCl solution (20 ml).

[0318] The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to dryness in vacuo to give the crude product, which was purified by flash column chromatography (SiO2, EtOAc / n-hexane = 30-45%) to afford Intermediate 31 as a white solid (190 mg, 37%).

[0319]

[0320] 1H NMR (400MHz, CD3OD): 6.67 (d, 1H, J = 7.6Hz), 6.64 (d, 1H, J = 2.0Hz), 6.51 (dd, 1H, J1 = 7.6Hz, J2 = 2.0Hz), 3.33 (t, 2H, J =7.2Hz),2.62(t,2H,J=7.2Hz),2.14(t,2H,J=7.6Hz),1.62-1.51(m,2H),1.37-1.22(m,24H),0.90(t,3H,J=6.8Hz);

[0321] 13C NMR(100MHz,CD3OD):178.3,146.3,144.8,132.0,121.0,116.8,116.3,42. 2,37.2,36.0,33.1,30.79,30.76,30.6,30.5,30.4,30.3,27.1,23.7,14.4.

[0322] Synthesis example of intermediate 33.

[0323] Under argon atmosphere, KCO (230 mg) and KI (43 mg) were added successively to a solution of amide 31 (170 mg; 0.43 mmol) and ethyl 4-bromobutyrate (32) (319 mg; 1.91 mmol) in anhydrous DMF (10 ml). The reaction thus obtained was stirred magnetically for 24 hours, then diluted with EtOAc (40 ml) and washed with HO (2×30 ml) and saturated aqueous NaCl solution (20 ml). The final organic phase was dried over anhydrous NaSO, filtered and concentrated to dryness under vacuum to give a crude product from which intermediate 33 (192 mg, 72%) was obtained as a white solid after purification by flash column chromatography.

[0324]

[0325] 1H NMR (400MHz, CDCl3): 6.81 (d, 1H, J = 5.6Hz), 6.75-6.65 (m, 2H), 5.47 (bt, 1H), 4 .13(q,4H,J=7.2Hz),4.013(t,2H,J=6.0Hz),4.007(t,2H,J=6.0Hz),4.04-3.9 8(m,4H),3.51-3.42(m,2H),2.72(t,2H,J=6.8Hz),2.53(t,4H,J=7.6Hz),2.16 -2.06(m,6H),1.64-1.53(m,2H),1.34-1.19(m,30H),0.87(t,3H,J=7.2Hz); 13C NMR(100MHz CDCl3):173.22,173.19,173.10,149.0,147.5,132.1,121.3,114.9,114.5,68.3,68.1,60.4,40.5,36.8, 35.2,30.70,30.68,29.7,29.62,29.59,29.5,29.34,29.32,29.29,25.7,24.73,24.70,22.7,14.2,14.1.

[0326] Synthesis Example of Intermediate 34

[0327] NaOH (66 mg) was added to a solution of ester 33 (58 mg; 0.095 mmol) in THF / MeOH / H2O=2 / 2 / 1 (5 ml): the reaction thus obtained was stirred for 18 hours and then carefully adjusted to pH about 5 by successive additions of small amounts of 2N HCl (780 μl in total). The reaction was then diluted with AcOEt (10 ml) and washed with H2O (2×6 ml) and saturated aqueous NaCl solution (6 ml). Finally, the organic phase was dried over anhydrous Na2SO4, filtered and concentrated to dryness in vacuo to give the diacid 34 as a white solid (49 mg, 91%). It appeared to be pure by NMR analysis and was used in the subsequent step without further purification.

[0328]

[0329] 1H NMR (400MHz, CD3OD): 6.82 (d, 1H, J = 8.4Hz), 6.78 (d, 1H, J = 1.6Hz), 6.71 (dd, 1H, J1 = 8.4Hz, J2 = 2.0Hz), 4.07-3.97 (m, 4H), 3.36 (t, 2H, J = 7.2Hz), 2. 70(t,2H,J=7.6Hz),2.52(t,2H,J=7.6Hz),2.51(t,2H,7.2Hz),2.16-2.01 (m,6H),1.60-1.50(m,2H),1.32-1.17(m,24H),0.86(t,3H,J=6.8Hz); 13C NMR(100MHz,CD3OD):176.6,175.7,149.7,148.2,133.3,122.2,115.84,115.48,69.22,68.98,4 1.5,36.9,35.6,32.5,31.1,30.28,30.26,30.1,29,995,29,968,29,863,26.6,25.4,23.2,14.3.

[0330] Synthesis Example of Intermediate 35

[0331] To a solution of 34 (50 mg; 0.089 mmol) in anhydrous DMF (3 ml) were added EDC·HCl (51 mg; 0.27 mmol) and, after 30 minutes, HOBt (26 mg; 0.31 mmol) under stirring and argon atmosphere. The reaction mixture was stirred for thirty minutes and then added dropwise to a solution of 16 (0.23 mol) and DIPEA (247 μl; 1.42 mmol) in anhydrous DMF (2 ml) under stirring at 0° C. (using an ice bath) and argon atmosphere via a syringe (over 5 minutes).

[0332] The reaction mixture thus obtained was slowly warmed to room temperature and then kept stirring for 24 hours, then diluted with EtOAc (20 ml) and washed successively with H2O (2 x 10 ml) and saturated aqueous NaCl solution (10 ml).

[0333] Finally the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to dryness under vacuum to give the crude product, which was purified by flash column chromatography (DCM / MeOH=1-3%) to give Intermediate 35 as a white solid (40 mg, 22%).

[0334]

[0335] 1H NMR (400MHz, CD3OD, TMS): 7.78, (m, 2H), 7.73 (dd, 2H, J1 = 8.8Hz, J 2＝ 2.4Hz),7.56-7.51(m,4H),7.49-7.24(m,36H),7.03(d,2H,J=8.8),6.74-6.60(m,3H),6.56(d,2H,J=1.6Hz),6. 45(d,2H,J=2.0Hz),5.24-5.17(m,12H),5.09(s,4H),4.07-4.0(m,4H),3.89(t,2H,J=6.4Hz),3.86(t,2H,J=6.4H z),3.63-3.57(m,4H),3.44(t,4H,J=3.2Hz),3.39-3.34(m,2H),3.34-3.28(m,4H),2.65(t,2H,J=7.2Hz),2.36-2 .27(m,4H),2.09(t,2H,J=7.6Hz),2,03-1.93(m,4H),1.61-1.50(m,2H),1.33-1.18(m,24H),0.87(t,3H,J=6.8);

[0336] 13 C NMR (100MHz, CD3OD):174.7,174.5,174.0,163.4,159.9,160.0,159.0,153.6,151.4,149.0,148.5,147.6,140.2,13 7.2,136.9,136.5,135.9,132.5,129.0,128.81,128.79,128.67,128.29,128.23,128.05,127.8,127.7,127.6,127. 0,123.7,123.2,121.6,116.0,115.2,114.8,114.1,109.9,98.4,94.3,72.0,71.5,71.2,71.0,70.8,70.2,69.7,68.8,68.6,40.9,39.5,36.7,35.2,32.70,32.65,32.1,29.9,29.7,29.60,29.56,29.53,26.1,25.63,25.58,22.9,14.2.

[0337] Synthesis example of compound 36

[0338] Under argon atmosphere, a solution of Pd / C (18 mg) and compound 35 (33 mg; 0.016 mmol) in EtOH / THF=2 / 1 (4 ml) was added sequentially to a 100 ml hydrogenation flask. Before introducing hydrogen (1 bar) into the flask, the reaction mixture was subjected to four nitrogen / vacuum cycles. The reactants were kept stirring at room temperature for 24 hours, then degassed under vacuum and filtered through 0.45 μm PTFE. The solution thus obtained was dried to dryness under vacuum: the crude product obtained as above was washed with a mixture of Et2O / n-hexane=2.5 / 7.5 (2×5 ml) and finally dried again in vacuum to obtain 36 as a yellow solid (13 mg, 60%).

[0339]

[0340] 1 H NMR (400MHz, CD3OD / MeOH=4 / 1, TMS): 7.74 (d, 2H, J = 1.6Hz), 7.54 (dd, 2H, J1 = 8.4Hz, J2 = 2.0Hz), 6.96-6.88 (m, 2H) 6.75-6.6 7(m,2H),6.62(dd,1H,J1=8.0Hz,J2=1.2Hz),6.39(d,2H,J=1.2Hz),6.25(dd,2H,J1=4.4Hz,J2=1.6Hz),4.08-4.01(m,4H), 3.98-3.88(m,4H),3.73-3.67(m,4H),3.57-3.49(m,4H),3.46-3.39(m,4H),3.39-3.35(m,2H),2.65(t,2H,J=7.2Hz),2.41 (t,4H,J=7.2Hz),2.15(t,2H,J=8.0Hz),2.11-2.01(m,4H),1.64-1.52(m,2H),1.37-1.20(m,24H),0.89,(t,3H,J=7.2Hz);

[0341] 13C NMR (100MHz, CD3OD / MeOH=4 / 1, TMS): 178.9, 175.2, 174.6, 164.5, 161.8, 157.3, 157.1 ,149.0,148.6,147.6,145.0,137.4,132.6,125.7,122.2,121.8,121.7,116.3,115.5, 115.2,114.8,105.3,99.3,94.3,71.8,70.2,69.9,68.8,68.7,57.9,41.1,39.7,36.8,35.3,33.0,30.5,30.0,29.9,29.8,29.7,29.62,29.57,26.2,25.8,22.9,18.0,14.2.

[0342] The compound comprises molecules (I) which can be linked to one another to obtain homodimers or heterodimers, trimers and tetramers according to formula (I) as described above, but with the exception of R1 or R1′, which consists of a linker L.

[0343] The thus modified formula (I) is referred to as formula (II), and is preferably as follows:

[0344]

[0345] X, Y, Z and a, b, c are as defined previously.

[0346] The linking group L is more preferably selected from the following compounds:

[0347]

[0348]

[0349]

[0350]

[0351]

[0352]

[0353]

[0354]

[0355]

[0356]

[0357]

[0358]

[0359]

[0360]

[0361]

[0362]

[0363] Most of the linkers shown in the table are known and commercially available. Any other linker can be readily prepared according to known methods.

[0364] 1. According to the general formula (I) or (II), the compound may have the following general formula (III), (IV), (V), (VI), (VII), (VIII), (IX), (X) and (XI):

[0365]

[0366]

[0367] Wherein d, e, f, g, h, i are each 0 to 12,

[0368] Wherein T1-T3, W1-W3 are each independently selected from: CH2, O, N(R1), S, NH, SO, SO2, OC(O), CO, NHC(O), C(O)NH, NH-C(O)-NH, NH-C(S)-NH.

[0369] 2. According to the general formula (I), the compound may have the following general formulae (XII), (XIII), (XIV) and (XV):

[0370]

[0371] Where n can independently take values ​​between 0 and 12;

[0372] R 10 The following may be independently but not exclusively, and any possible combination of the following may be:

[0373] -hydrogen;

[0374] -C 1-24 Alkyl or heteroalkyl, C 1-24 Alkenyl or heteroalkenyl; C 1-24 Alkynyl or heteroalkynyl;

[0375] - acyl residues of saturated / unsaturated / polyunsaturated fatty acids of synthetic or natural origin;

[0376] - a residue which may preferably, but not exclusively, be selected from the residues shown in the following table:

[0377]

[0378]

[0379] Below are some examples of compounds according to the invention, the technical effect of which has been demonstrated.

[0380]

[0381]

[0382]

[0383]

[0384]

[0385] The compound can also be prepared by the method described in patent application WO-A-99 / 66062 (National Research Council and Felice Rao's Rao-Erbe). In this patent, it is described from page 3, line 8 to page 9, line 22, which is hereby incorporated by reference. However, other similar methods can be used.

[0386] Therefore, the present invention also defines a novel method for treating the above tumors using the above compounds, and a novel method for preparing a drug for treating the above tumors.

[0387] The compounds according to the invention, in particular the synthetic quercetin derivatives described and illustrated herein, were studied in vitro using human tumor cells and evaluating their effects on the tumor proliferation capacity of the above tumors. Specifically, the following human tumor cell lines were experimentally evaluated: 1) HCT116, colon adenocarcinoma cells; 2) H1299, lung adenocarcinoma cells; 3) A375, melanoma cells; and 4) A431, squamous cell carcinoma of the skin; 5) 786-O, renal carcinoma cells; 6) PC3, prostate cancer cells; 7) BT549, breast cancer cells; 8) CAL27, tongue cancer cells (belonging to head and neck cancer); 9) HepG2, hepatocellular carcinoma cells; 10) PANC-1, pancreatic cancer cells; 11) T24, bladder cancer cells; 12) OVCAR-3, ovarian cancer cells. In addition, a normal human cell line, WS1 (skin fibroblasts), was also treated with the novel synthetic compounds.

[0388] Cell proliferation was quantified by the crystal violet method using a spectrophotometer. Briefly, after acquiring serial images (0, 24, and 48 h) using an EVOS XL Cell Imaging System microscope (Thermo Fisher Scientific), cells imaged at 48 h were fixed with paraformaldehyde (4%) and stained with crystal violet (1%). Then, the cells were analyzed using a spectrophotometer ( 200PRO, Tecan) measured the specific absorbance (λ = 590nm) at 48 hours. Data analysis was performed using a dedicated Excel file for graphical representation and statistical processing (t-test). In addition, the results obtained using crystal violet were confirmed by using another cell proliferation assessment method based on the real-time cell analyzer XCELLigenceRTCA DP (ACEA Biosciences, Inc.). This tool allows the growth of cells to be monitored based on the quantitative value (so-called cell index) derived from the electrical impedance measured by the tiny electrodes at the bottom of the culture well. By setting the cell index measured every minute on the instrument, proliferation can be monitored for several days (5-7).

[0389] The data were analyzed by processing various parameters such as cell replication time, slope and maximum cell index calculated by dedicated software.

[0390] The results show that the compounds of the present invention, in particular the various compounds described and illustrated herein, significantly inhibited the proliferation of adenocarcinoma cells of the colon (HCT116) and lung (H1299), melanoma cells (A375) and squamous cell carcinoma of the skin (A431), renal carcinoma (786-O), prostate carcinoma (PC3), breast carcinoma (BT549), tongue carcinoma (CAL27), hepatocellular carcinoma (HepG2), pancreatic carcinoma (PANC-1), bladder carcinoma (T24) and ovarian carcinoma (OVCAR-3) compared to the control (treated with solvent vehicle only).

[0391] Tumor cell apoptosis was quantified by the acridine orange / ethidium bromide (AO / EB) method using fluorescence microscopy and Image J. In brief, after treatment with the test compound (2 hours), the tumor cells were contacted with AO / EB solution (in sterile H2O) and photographed under a fluorescence microscope (Leica) using a ≥20x objective (to distinguish the morphology of the nucleus). In this way, live cells are labeled with AO and fluoresce in the green channel, while necrotic cells that have undergone plasma membrane rupture can pass through EB and fluoresce in the red channel. In contrast, apoptotic cells are permeable to both AO and EB, so when the fluorescence obtained by two separate microscopes is superimposed using the editing program Image J, the colocalization of the two fluorescences is displayed as yellow-orange. Objective quantification of colocalized areas (apoptotic cells) and non-colocalized areas (live cells or necrotic cells) was performed under conditions where the number of cells was greater than 300 / , and was displayed as the percentage of viable (green), necrotic (red) or apoptotic (yellow-orange) tumor cells. Data analysis was performed by using GraphPad Prism 6 for graphical representation and statistical processing (two-way ANOVA test).

[0392] The compounds according to the invention were compared with simple quercetin. In these comparisons, the dose response curves showed that the compounds according to the invention had a significantly more powerful and effective effect in inhibiting the proliferation and inducing apoptosis of all tested tumor cells of the above-mentioned type.

[0393] Furthermore, the compounds of the present invention induce cell death and apoptosis in colon adenocarcinoma (HCT116) and melanoma (A375) cells in a very short time (<2 hours) compared to the control (treated with solvent vehicle only). Very importantly, the compounds of the present invention, in particular all the compounds described and illustrated herein, are significantly less efficient in inhibiting the proliferation of normal human cells (WS1 skin fibroblasts) and are unable to induce apoptosis compared to tumor cells, indicating that the toxic effects of the patented compounds may be clinically selective, acting only on tumors while sparing the patient's healthy cells.

[0394] The progression of the tumor cell cycle was assessed by the propidium iodide method, and the cellular DNA content was quantified using a cell fluorometer. In brief, after treatment with the test compound (in duplicate) (24 hours), the tumor cells were washed (with PBS), dissociated with trypsin-EDTA, and precipitated by centrifugation (125g, 6 minutes). The cells were then fixed in ethanol (70%) and stored in a freezer (-20°C) for 1-3 days. After further washing (PBS) and treatment with RNase (50μg / ml), the cells were contacted with propidium iodide (50μg / ml) (protected from light, ice bath; 30 minutes). FL2 and FL3 cell fluorometer readings analyzed approximately 50,000 cells per sample (1μl / second). Statistical analysis and graphical representation were performed using t-tests and preset Excel property files.

[0395] Intracellular ROS levels in tumor cells were measured using the probe 2,7-dichlorodihydrofluorescein diacetate (DCFH2-DA) and a fluorescence spectrophotometer. DCFH2-DA easily diffuses through the cell membrane into the cytoplasmic space of the cell, where it is hydrolyzed by intracellular esterases into an impermeable DCFH2 product. DCFH2 is oxidized by intracellular ROS (mainly H2O2, HO·, ROO·, NO·, and ONOO), resulting in the production of intracellular fluorescent DCF, whose concentration is proportional to the level of ROS in the cell. Once the treatment with the test compound is completed (1 hour), the tumor cells are washed (with PBS), the probe DCFH2-DA (10 μM) is added after 30 minutes, and after further washing in PBS, the intracellular DCF fluorescence is quantified by using a spectrophotometer (Synergy; emitting at 498nm and 530nm). All experiments were repeated at least 3 times. Data analysis was performed using GraphPad Prism 6 for graphical representation and statistical processing (one-way AN OVA test).

[0396] The results show that the compounds according to the invention terminate the replicative tumor cell cycle. In fact, compared with the control treated with the solvent vehicle, the tumor cells (particularly colon cancer and melanoma cells) treated with the compounds of the invention accumulate significantly in the S phase and G2 phase of the cell cycle. This effect on the cell cycle is partly mediated by ROS, which is also confirmed by the fact that the compounds according to the invention promote a rapid increase in the level of intracellular ROS in malignant tumor cells (<1h). Therefore, the purpose of the synthetic flavonoid derivatives of the present invention is to selectively inhibit the growth and proliferation of malignant epithelial tumors at least by promoting cell death and apoptosis processes, terminating the cell cycle and generating intracellular ROS.

[0397] In detail, the medicinal compounds according to the present invention exhibited a greater antiproliferative effect in melanoma than the parent chemical skeleton. Figure 1aAs shown, the proliferation of human melanoma cells (A375) was quantified using the crystal violet method and a spectrophotometer. Quercetin (Que) and its synthetic derivatives (compounds 2, 4, and 5) were compared ( Figure 1a ; 60, 60 μM). Compounds 2, 4 and 5 significantly inhibited the growth of melanoma cells, and their efficiency was higher than that of quercetin ( Figure 1b ). Ctr, control (treated with vehicle). *, p<0.05; **, p<0.01; and ***, p<0.001 compared with control by Student's t-test.

[0398] Synthetic flavonoid derivatives showed greater antiproliferative effects than the parent chemical skeleton in cutaneous squamous cell carcinoma (SCC). The proliferation of SCC cells (A431) was quantified by using the crystal violet method and spectrophotometry. Quercetin (Que) and its synthetic derivatives (compounds 2, 4, and 5) ( Figure 2a ; 90, 90 μM). Quercetin had no effect, while compounds 2, 4 and 5 significantly inhibited the growth of A431 cells ( Figure 2b ). Ctr, control (treated with vehicle). **, p<0.01 compared with control by Student's t-test.

[0399] Synthetic flavonoid derivatives exhibited greater antiproliferative effects than the parent chemical skeleton in colorectal cancer. The proliferation of human colon cancer cells (HCT116) was quantified using the crystal violet method and spectrophotometer. Quercetin (Que) and its synthetic derivatives (Compound 2, Compound 3, Compound 4, and Compound 5) ( Figure 3a ; 100, 100 μM). Compounds 2, 3, 4 and 5 significantly inhibited the growth of HCT116 cells ( Figure 3b ). Ctr, control (treated with vehicle). *, p<0.05; **, p<0.01 compared with control by Student's t-test.

[0400] Synthetic flavonoid derivatives showed greater antiproliferative effects than the parent chemical skeleton in renal cancer. The proliferation of human renal cancer cells (786-O) was quantified using the crystal violet method and spectrophotometer. Quercetin (Que) and its synthetic derivatives (Compound 1, Compound 2, Compound 3, Compound 4 and Compound 5) ( Figure 4a ; 80, 80 μM). Quercetin had no effect, while compounds 1, 2, 3, 4 and 5 significantly inhibited the growth of 786-O cells ( Figure 4b). Ctr, control (treated with vehicle). *, p<0.05; **, p<0.01; and ***, p<0.001 compared with control by Student's t-test. Synthetic flavonoid derivatives exhibited greater antiproliferative effects in prostate cancer than the parent chemical skeleton. Proliferation of human prostate cancer cells (PC3) was quantified by using the crystal violet method and spectrophotometry. Quercetin (Que) and its synthetic derivatives (Compound 1, Compound 2, Compound 3, Compound 4, and Compound 5) ( Figure 5a ; 80, 80 μM). Quercetin showed moderate antiproliferative effects, while compounds 1, 3, 4, and 5 were better and significantly inhibited the growth of PC3 cells ( Figure 5b ). Ctr, control (treated with vehicle). *, p<0.05; and **, p<0.01 compared with control by Student's t-test.

[0401] Synthetic flavonoid derivatives showed greater antiproliferative effects than the parent chemical skeleton in breast cancer. The proliferation of human breast cancer cells (BT549) was quantified spectrophotometrically using the crystal violet method. Quercetin (Que) and its synthetic derivatives (Compound 1, Compound 2, Compound 3, Compound 4 and Compound 5) ( Figure 6a ; 80, 80 μM). Compounds 1, 2, 3, 4 and 5 significantly inhibited the growth of BT549 cells ( Figure 6b ). Ctr, control (treated with vehicle). *, p<0.05; **, p<0.01 compared with control by Student's t-test.

[0402] Synthetic flavonoid derivatives showed greater antiproliferative effects than the parent chemical skeleton in tongue cancer (head and neck cancer, HNC). The proliferation of human tongue cancer cells (Cal27) was quantified spectrophotometrically using the crystal violet method. Quercetin (Que) and its synthetic derivatives (Compound 1, Compound 2, Compound 3, Compound 4 and Compound 5) ( Figure 7a ; 80, 80 μM). All synthesized compounds significantly inhibited tumor growth, and the antiproliferative effect of compound 5 was superior to that of quercetin ( Figure 7b ). Ctr, control (treated with vehicle). *, p<0.05; **, p<0.01; and ***, p<0.001 compared with control by Student's t-test.

[0403] Synthetic flavonoid derivatives showed greater antiproliferative effects in liver cancer than the parent chemical skeleton. The proliferation of human hepatocellular carcinoma cells (HepG2) was quantified spectrophotometrically using the crystal violet method. Quercetin (Que) and its synthetic derivatives (Compound 1, Compound 2, Compound 3, Compound 4 and Compound 5) ( Figure 8a; 80, 80 μM). All synthesized compounds could significantly inhibit tumor growth, and the anti-proliferative effects of compounds 3, 4 and 5 were superior to those of quercetin ( Figure 8b ). HepG2. Ctr, control (treated with vehicle). *, p<0.05; **, p<0.01 compared with control by Student's t-test.

[0404] Synthetic flavonoid derivatives showed greater antiproliferative effects in pancreatic cancer than the parent chemical skeleton. The proliferation of human pancreatic cancer cells (PANC-1) was quantified spectrophotometrically using the crystal violet method. Quercetin (Que) and its synthetic derivatives (Compound 1, Compound 2, Compound 3, Compound 4 and Compound 5) ( Figure 9a ; 80, 80 μM). Although quercetin had no effect, compounds 2, 3, 4, and 5 significantly inhibited the growth of PANC-1 cells ( Figure 9b ). Ctr, control (treated with vehicle). *, p<0.05; **, p<0.01 compared with control by Student's t-test.

[0405] Synthetic flavonoid derivatives showed greater antiproliferative effects than the parent chemical skeleton in bladder cancer. The proliferation of human bladder cancer cells (T24) was quantified spectrophotometrically using the crystal violet method. Quercetin (Que) and its synthetic derivatives (Compound 1, Compound 2, Compound 3, Compound 4 and Compound 5) ( Fig.10a ; 80, 80 μM). Although quercetin had no effect, compounds 2, 3, 4, and 5 significantly inhibited the growth of T24 cells ( Fig.10b ). Ctr, control (treated with vehicle). *, p<0.05; **, p<0.01; and ***, p<0.001 compared with control by Student's t-test.

[0406] Synthetic flavonoid derivatives showed greater antiproliferative effects than the parent chemical skeleton in ovarian cancer. The proliferation of human ovarian cancer cells (OVCAR-3) was quantified spectrophotometrically using the crystal violet method. Quercetin (Que) and its synthetic derivatives (Compound 5, Compound 19c, Compound 19d, Compound 29e, and Compound 29f) ( Fig.11a ; 60, 60 μM). Although quercetin had no effect, compounds 5, 19c, 19d, 29e and 29f significantly inhibited the growth of OVCAR-3 cells ( Fig.11b ). Ctr, control (treated with vehicle). *, p<0.05; **, p<0.01; and ***, p<0.001 compared with control by Student's t-test.

[0407] The synthetic flavonoid derivative compound 26 exhibited greater antiproliferative effects in melanoma than the parent chemical skeleton. The proliferation of human melanoma cells (A375) was quantified spectrophotometrically using the crystal violet method. Compound 26 significantly inhibited the proliferation of A375 cells with higher potency compared to quercetin. Ctr, control (treated with vehicle). *, p<0.05; **, p<0.01 ( Fig.12 ).

[0408] Synthetic flavonoid derivatives, compound 26 and compound 27, showed greater antiproliferative effects in colorectal cancer than the parent chemical skeletons. The proliferation of human colon cancer cells (HCT116) was quantified using the crystal violet method and spectrophotometry. Quercetin (Que) and its synthetic derivatives (compounds 26 and 27) ( Fig.13a ; 100, 100 μM). Compared with quercetin, compound 26 significantly inhibited the growth of HCT116 cells ( Fig.13b ), with higher efficacy. Ctr, control (treated with vehicle). *, p<0.05; **, p<0.01 compared with control by Student's t-test.

[0409] The synthesized flavonoid derivatives exhibited significant antiproliferative effects in melanoma. The proliferation of human melanoma cells (A375) was quantified spectrophotometrically using the crystal violet method. The synthetic compounds used at concentrations of 10 μM and 30 μM significantly inhibited the growth of A375 cells with high potency. Ctr, control (treated with vehicle). 15a, compound 15a; 15b, compound 15b; 15c, compound 15c; 15d, compound 15d; 19b, compound 19b; 19c, compound 19c; 19d, compound 19d; 29a, compound 29a; 29c, compound 29c; 29d, compound 29d; 29e, compound 29e; 29f, compound 29f; and 29g, compound 29g. *, p<0.05; **, p<0.01; ***, p<0.001 (compared with control by Student's t-test). Fig.14 ).

[0410] Synthetic flavonoid derivatives exhibit significant antiproliferative effects in colorectal cancer. The proliferation of human colon cancer cells (HCT116) was quantified using the crystal violet method and spectrophotometer. The synthetic compounds used at concentrations of 10 μM and 30 μM significantly inhibited the growth of HCT116 cells with high potency. Ctr, control (treated with vehicle). 15a, compound 15a; 15b, compound 15b; 15c, compound 15c; 15d, compound 15d; 19b, compound 19b; 19c, compound 19c; 19d, compound 19d; 29a, compound 29a; 29c, compound 29c; 29d, compound 29d; 29e, compound 29e; 29f, compound 29f; and 29g, compound 29g. *, p<0.05; **, p<0.01; ***, p<0.001 (compared with control by Student's t-test). Fig.15 ).

[0411] Synthetic trimeric flavonoid derivatives exhibit antiproliferative effects in colorectal cancer and melanoma. The proliferation of human colon cancer (HCT116) and melanoma (A375) cells was quantified spectrophotometrically using the crystal violet method. Synthetic trimeric compounds (compound 22a and compound 36) used at concentrations of 30 μM and 100 μM inhibited the growth of HCT116 and A375 cells. Ctr, control (treated with vehicle). 22a, compound 22a; 36, compound 36. *, p<0.05 (compared with control by Student's t-test) Fig.16 ).

[0412] Synthetic flavonoid derivatives do not interfere with normal human cell proliferation. The proliferation of human skin fibroblasts (WS1) was studied using crystal violet and spectrophotometry. Quercetin (Que), its synthetic derivative (Compound 2) and apigenin (API) were investigated at the indicated concentrations. Apigenin significantly inhibited the proliferation of normal human skin cells, while quercetin and compound 2 did not interfere with their growth. Ctr, control (treated with vehicle). *, p<0.05; ***, p<0.005 (compared with control by Student's t-test). Fig.17 ).

[0413] Synthetic flavonoid derivatives selectively induce apoptosis in tumor cells. The percentage of cells that initiate the cell death program (apoptosis) was quantified by the acridine orange / ethidium bromide method and using fluorescence microscopy and Image-J. All synthetic quercetin derivatives (compounds 2, compound 5, compound 4, compound 3, and compound 1) induced apoptosis (at 2 hours) in human colon (HCT116) and skin (melanoma, A375) tumor cell lines, but did not induce apoptosis in normal cells (human skin fibroblasts, WS1). The following concentrations of substances were used (relative to their respective antiproliferative IC50): compound 2, 50 μM (HCT116) or 40 μM (A375, WS1); compound 5, 40 μM; compound 4, 40 μM; compound 3, 60 μM; compound 1, 60 μM (HCT116) or 80 μM (A375, WS1). Ctr, control (treated with vehicle). Compared with the control group, *, p<0.05; **, p<0.01; ***, p<0.005 and ****, p<0.001 were tested by two-way ANOVA. Fig.18 ).

[0414] Synthetic flavonoid derivatives terminate the cell cycle of melanoma. The evolution of human melanoma cells (A375) throughout the cell cycle was quantified by using the propidium iodide (PI) method and a cell fluorometer. Quercetin (Que) and its synthetic derivatives (Compound 1, Compound 2, Compound 3, Compound 4 and Compound 5) were used at their respective antiproliferative IC50s. With the exception of Compound 3, all investigated compounds significantly terminated tumor cells in S and / or G2. Ctr, control (treated with vehicle). H2O2, hydrogen peroxide (positive control; 200 μM). *, p<0.05; **, p<0.01; ***, p<0.001 (compared with control by Student's t-test). Fig.19 ).

[0415] Synthetic flavonoid derivatives can terminate the cell cycle of colorectal cancer. The evolution of human colon cancer cells (HCT116) throughout the cell cycle was quantified by using the propidium iodide (PI) method and a cell fluorometer. Quercetin (Que) and its synthetic derivatives (Compound 1, Compound 2, Compound 3, Compound 4 and Compound 5) were used at their respective antiproliferative IC50s. Compounds 1, 3 and 4 significantly terminated tumor cells in S. Ctr, control (treated with vehicle). H2O2, hydrogen peroxide (positive control; 200 μM). *, p<0.05; **, p<0.01; ***, p<0.001 ( Fig. 20 ).

[0416] The effect of compound 1 on the tumor cell cycle is partially mediated by ROS. The cell cycle of human colon cancer (HCT116) and melanoma (A375) cells was investigated using the propidium iodide method and a cell fluorimeter. The reactive oxygen species (ROS) scavenger, N-acetylcysteine ​​(NAC; 10 mM), at least partially blocked the effects of compound 1 (used at its antiproliferative IC50) induced on the cell cycle in HCT116 (left) and A375 (right) cells. Ctr, control (treated with vehicle). By Student's t-test, compared with the respective controls (compound 1 for compound 1+NAC, compound 1), *, p<0.05; **, p<0.01; ***, p<0.001 ( Fig.21 ).

[0417] Synthetic flavonoid derivatives cause elevated intracellular ROS levels in colorectal cancer. Intracellular reactive oxygen species (ROS) levels in human colon cancer cells (HCT116) were quantified 2 h after treatment using a fluorescent DCFH2-DA probe. Fluorescence data were measured with a spectrophotometer and expressed in arbitrary units (AU). Quercetin (QUE) and its synthetic derivatives (Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, and Compound 19b) were used with their respective antiproliferative IC50s. Quercetin had no effect, while Compound 1, Compound 2, Compound 3, and Compound 19b significantly increased ROS levels in HCT116 cells (left). The effect of Compound 1 was blocked by pretreatment with the ROS scavenger N-acetyl-L-cysteine ​​(NAC; 10 mM) (right). H2O2, hydrogen peroxide (positive control; 500 μM). Ctr, control (treated with vehicle). By one-way ANOVA test, *, p<0.05; **, p<0.01; ***, p<0.001 compared with the control ( FIG. 22 ).

[0418] The following table is divided into two parts, which shows the inhibitory concentration (in μM) representing 50% of the maximum antiproliferative effect (IC50) corresponding to each compound of the present invention (whose number is shown in the first column on the left) when tested on each human malignant tumor cell line (row 1, starting from the second column). Cell proliferation was quantified spectrophotometrically by the crystal violet method.

[0419]

[0420]

[0421]

[0422]

[0423]

[0424] Legend: CRC, colorectal cancer; LC, lung cancer; SCC, squamous cell carcinoma; PCC, pancreatic cancer; RCC, renal cell carcinoma; PC, prostate cancer; BC, bladder cancer; BRC, breast cancer; HNC, head and neck cancer; HCC, hepatocellular carcinoma; OCC, ovarian cancer; ND, not determined.

Claims

1. A medicinal compound selected from:

2. A medicinal compound selected from:

3. Use of the compound according to claim 1 or 2 in the preparation of a medicament for treating lung cancer.

4. Use of the compound according to claim 1 or 2 in the preparation of a medicament for treating colon cancer.

5. Use of the compound according to claim 1 or 2 in the preparation of a medicament for treating melanoma.

6. Use of the compound according to claim 1 or 2 in the preparation of a medicament for treating malignant epithelial tumors.

7. Use of the compound according to claim 1 or 2 in the preparation of a medicament for treating cutaneous squamous cell carcinoma.

8. Use of the compound according to claim 1 or 2 in the preparation of a medicament for treating basal cell carcinoma or basal cell tumor.

9. Use of the compound according to claim 1 or 2 in the preparation of a medicament for treating one or more of the following malignant tumors: head and neck cancer, gastric cancer, liver cancer, pancreatic cancer, breast cancer, prostate cancer, bladder cancer, kidney cancer, mesothelioma and ovarian cancer.

10. Use of a compound according to claim 1 or 2 in the preparation of a medicament for treating one or more of the following malignant tumors: salivary gland tumors, esophageal cancer, small intestinal tumors, gallbladder cancer and extrahepatic bile duct cancer, soft tissue sarcomas, cervical cancer, uterine cancer, testicular cancer, urinary tract tumors, choroidal melanoma, thyroid cancer, endometrial cancer, retinal tumors, uveal melanoma, anal canal cancer and anal cancer, bone and joint cancer.

11. Use of a compound according to claim 1 or 2 as a monotherapy or in combination with radiotherapy, chemotherapy, hormone therapy, biological therapy, immunotherapy and / or gene therapy in the preparation of a medicament for the treatment of cancer patients.

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