WO2021234468A1

WO2021234468A1 - Curcumin dihydropyridone derivatives with anti-cancer activity

Info

Publication number: WO2021234468A1
Application number: PCT/IB2021/052354
Authority: WO
Inventors: Rufus SMITS
Original assignee: Latvian Institute Of Organic Synthesis
Priority date: 2020-05-18
Filing date: 2021-03-22
Publication date: 2021-11-25
Also published as: DK202270604A1; CA3177353A1; LV15600A

Abstract

The present invention includes DHPDO curcumin derivatives (6) having the following general structure: These DHPDO curcumin derivatives have pronounced anti-cancer activity with lower toxicity on normal cells. Specifically, these compounds can be used in the treatment of breast cancer and colorectal cancer.

Description

Curcumin dihydropyridone derivatives with anti-cancer activity Description of the invention [001] The invention relates to medicine, namely to curcumin dihydropyridone derivatives, in particular to (E) -4-phenyl-7-styryl-3,4-dihydro-2H-pyrano [4,3-b] pyridine-2,5 ( 1H) -dione (6a) and (E)-7-(4-hydroxy-3-methoxystyryl)-4-phenyl-3,4-dihydro-2H-pyrano[4,3-b]pyridine- 2,5(1H)-dione (6b). [002] The novel compounds belong to derivatives of natural substances with pharmaceutical and anti-cancer activity for the treatment of cancer. The compounds belong to a class of anticancer agents against breast cancer and colorectal cancer. Background of the art [003] Breast cancer is the most common invasive cancer, and colorectal cancer is the second most commonly diagnosed malignancy in women worldwide. For men, colorectal cancer is the third most common cancer. Despite recent advances in targeted therapy, clinical efficacy is limited, incurable, with a high toxicity profile and excessive cost. Therefore, there is a growing interest in identifying safe and available natural compounds as an adjunct to the traditional therapies currently offered to these patients. [004] Curcumin from the roots of Curcuma longa is one such compound that has become one of the leading and most studied natural agents due to its role in cancer prevention. However, its rapid metabolism, low bioavailability, and lack of specificity require the synthesis of new curcumin analogs with increased potency and higher specificity. [005] Breast cancer is the most common and second leading cause of cancer-related deaths among women in the world [1,2], with 2 million new cases and 627,000 deaths at the end of 2018 [3]. Colorectal cancer, on the other hand, is the third most common cancer in men and the second most common cancer diagnosed in women, with more than 1.8 million new cases and approximately 861,000 deaths reported in 2018 [4]. [006] Current breast cancer treatment strategies include surgery, chemotherapy, and radiation therapy, but may lack efficacy due to a high risk of relapse, poor patient response, and the emergence of drug resistance [5]. The prognosis of colorectal cancer varies with the stage of the cancer, with almost half of patients undergoing medical surgery alone and another 20-25% of patients receiving postoperative adjuvant chemotherapy having cancer recurrence, metastasis, and possible death [6-8], emphasizing the inadequacy of treatment options for this fatal malignancy in the current situation [9]. Therefore, there is a growing interest in identifying safe and available natural compounds as an adjunct to the traditional therapies currently offered to these patients. [007] Curcumin from the roots of Curcuma longa L., with its role in cancer prevention, has become one of the leading and most studied natural remedies. Curcumin is a polyphenol phytochemical compound 1,7-bis (4-hydroxy-3-methoxyphenyl) -1,6-heptadiene-3,5-dione [10,11] that interacts with many biological targets, including inflammatory mediators, growth factors , enzymes, carrier proteins, metal ions, tumor suppressors, transcription factors, oncoproteins and cellular nucleic acids [12 - 14]. [008] Curcumin has been consumed in Asian countries for more than 2,000 years due to its various therapeutic properties against human diseases, including cancer and autoimmune diseases [15-23]. However, despite the potential for cancer prevention and treatment, supported by many preclinical studies and even clinical trials, there are concerns about selectivity and bioavailability of curcumin after oral administration. Poor bioavailability of curcumin is attributed to poor absorption, rapid metabolism, and systemic elimination, as well as interactions with many molecular targets, reducing selectivity and inhibiting its use as a therapeutic agent [24, 25]. [009] To overcome the practical limitations of curcumin bioavailability, several approaches have been adopted to improve its systemic absorption [26, 27]. Great efforts have also been made to synthesize new curcumin derivatives [28, 29] in order to optimize the anti-cancer properties of curcumin and to improve its pharmacokinetic profile [30]. In this invention curcumin derivatives are based on the 3,4-dihydro-2 (1H) -pyridone (DHPDO) heterocycle, which has significant pharmaceutical efficacy and a spectrum of therapeutic activities, such as antibacterial, antifungal, antitumor and HIV-1 specific reverse transcriptase inhibitors [ 31]. DHPDO curcumin derivatives provide even more diverse and improved pharmacological properties. Aim of the invention [010] The present invention provides two curcumin dihydropyridone derivatives (6a) and (6b) with significant anti-cancer activity on breast and colorectal cancer cell lines and at the same time with much lower toxicity on normal cells. Detailed description of the invention [011] DHPDO curcumin derivatives are synthesized according to the general scheme (Scheme 1):

where R₁=3: H, OH, OCH₃, N(CH₃)₂ in combination with R₂=3: H, OH, OCH₃, N(CH₃)₂ R₁=4: H, OH, OCH₃, N(CH₃)₂ in combination with R₂=4: H, OH, OCH₃, N(CH₃)₂ R₁=3,4: (OCH₃, OH), (OH, OH), (OCH₃, OTHP), (OCH₃, O(CH₂)_nN(CH₂CH₃)₂ in combination with R₂=3,4: (OCH₃, OH), (OH, OH), (OCH₃, OTHP), (OCH₃, O(CH₂)_nN(CH₂CH₃)₂, optical isomers, polymorphs and pharmaceutically acceptable addition salts, hydrates and solvates thereof. Scheme 1. Reaction of 4-amino-6-methyl-2H-pyran-2-one (1) with substituted benzaldehyde (2) and Meldrum's acid (3) in a boiling acid-catalyzed solvent (e.g., ethyl alcohol) affords DHPDO pyranone derivatives (4) . Further, the reaction of these derivatives with substituted benzaldehydes (5) in a dry solvent (eg methanol) with a base (eg magnesium methoxide) yields curcumin derivatives of DHPDO (6). Examples of implementation of the invention Synthesis example I [013] 7-Methyl-phenyl-3,4-dihydro-2H-pyrano [4,3-b] pyridine-2,5 (1H) -dione (4a). Weigh 0.89 g (0.71 mmol) of 4-amino-6-methyl-2H-pyran-2-one, 1.03 g (0.71 mmol) of Meldrum's acid into a 50 mL round bottom flask with magnetic stirrer, and 0.8 g (0.75 mmol) of benzaldehyde. Add 20 ml of ethanol and 0,05 g of p-toluenesulphonic acid into the flask. The mixture is refluxed for 18 hours, then cooled to room temperature and poured into ice water. The precipitate is washed with water and, after drying, 1.12 g of a white solid are obtained in a yield of 63%. [014] C₁₅H₁₃NO₃ MW. 255,27; LC/MS: ESI [M+] 256. Elemental analysis: C 70,58; H 5,13; N 5,49; found: C 70,46; H 5,06; N 5,41. [015] ¹H NMR (400 MHz, DMSO-d6) δ 10,63 (s, 1H), 7,35 – 7,25 (m, 2H), 7,25 – 7,17 (m, 1H), 7,17 – 7,10 (m, 2H), 6,04 (d, J = 1,1 Hz, 1H), 4,16 (d, J = 7,9 Hz, 1H), 3,12 (dd, J = 16,5; 8,1 Hz, 1H), 2,58 – 2,51(m, 1H), 2,22 (d, J = 0,9 Hz, 3H). [016] ¹³C NMR (101 MHz, DMSO-d6) δ 169,87; 162,45; 161,92; 149,55; 142,12; 128,73; 126,90; 126,40; 98,72; 97,63; 37,97; 34,86; 19,58. Synthesis example II [017] 2-Methoxy-4- (7-methyl-2,5-dioxo-1,3,4,5-tetrahydro-2H-pyrano [4,3-b] pyridin-4-yl) phenylacetate (4b). Using the procedure of Example I, but substituting 4-vanillin acetate for benzaldehyde, a white solid was obtained in 68% yield. [018] C₁₈H₁₇NO₆ MW. 343,34 LC/MS:ESI [M+] 344. Elemental analysis: C 62,97; H 4,99; N 4,08; found: C 62,91; H 5,02; N 4,03. [019] ¹H NMR (400 MHz, DMSO-d6) δ 10,65 (s, 1H), 7,02 (d, J = 2,0 Hz, 1H), 6,97 (d, J = 8,1 Hz, 1H), 6,58 (dd, J = 8,2; 2,0 Hz, 1H), 6,04 (d, J = 1,0 Hz, 1H), 4,18 (d, J = 7,8 Hz, 1H), 3,73 (s, 3H), 3,12 (dd, J =16,5; 8,1 Hz, 1H), 2,61 (dt, J = 16,3; 1,3 Hz, 1H), 2,23 (s, 3H), 2,22 – 2,19 (m, 3H). [020] ¹³C NMR (101 MHz, DMSO-d6) δ 169,92; 168,59; 162,56; 161,96; 150,88; 149,65; 140,98; 138,18; 122,80; 117,43; 111,67; 98,54; 97,65; 55,64; 37,89; 34,76; 20,37; 19,59. Synthesis example III [021] (E)-4-Phenyl-7-styryl-3,4-dihydro-2H-pyrano[4,3-b]pyridine-2,5-(1H) -dione (6a). Weigh 100 mg of magnesium turnings and 20 ml of dry methanol into a 50 ml round-bottomed flask with a magnetic stirrer. Heat the flask in an oil bath, after all the magnesium has dissolved, add 7-methyl-phenyl-3,4-dihydro-2H-pyrano[4,3-b]pyridine-2,5-(1H)-dione (4a) 0.26 g (1.0 mmol) and 0.11 g (1.04 mmol) of benzaldehyde. The reaction mixture is boiled for 18 hours. The reaction mixture is cooled and the precipitate is filtered off, 0.18 g of a pale yellow powder is obtained in a yield of 53%. [022] C₂₂H₁₇NO₃ MW 343,38 LCMS: ESI [M+] 344.34. Elemental analysis: C 76,95; H 4,99; N 4,08; found: C 76,05; H 5,04; N 4,18. [023] ¹H NMR (400 MHz, Chloroform-d) δ 8,57 (s, 1H), 8,39 (s, 1H), 7,48 (s, 1H), 7,42 (t, J = 1,6 Hz, 2H), 7,36 – 7,26 (m, 4H), 7,26 – 7,12 (m, 12H), 6,54 (d, J = 16,0 Hz, 1H), 5,86 (s, 1H), 5,70 (s, 1H), 4,35 (ddd, J = 26,8; 8,1; 1,6 Hz, 2H), 3,00 (ddd, J = 16,9; 14,6; 8,2 Hz, 2H), 2,93 – 2,74 (m, 2H), 2,18 (s, 3H). Synthesis example IV [024] (E)-7-(4-hydroxy-3-methoxystyryl)-4-phenyl-3,4-dihydro-2H-pyrano[4,3-b]pyridine- 2,5(1H)-dione (6b). [025] The reaction proceeds according to Synthesis Example IV, but tetrahydropyranyl-protected vanillin (1.2 mol eq) is added instead of benzaldehyde. After boiling for 18 hours, the mixture was cooled and dilute hydrochloric acid (1 mL) was added. The precipitate is filtered off and washed with water. 0.09 g of a yellow substance is obtained in a yield of 43%. [026] C₂₃H₁₉NO₅ MW. 389,41 LC/MS: ESI [M+] 390.43 Elemental analysis: C, 70,94; H, 4,92; N, 3,60; found: C, 70,83; H, 5,01; N, 3,65. [027] ¹H NMR (400 MHz, Hloroforms-d) δ 7,35 (dt, J = 15,9; 2,5 Hz, 1H), 7,28 (dd, J = 3,2; 1,9 Hz, 2H), 7,19 (tdd, J = 10,7; 6,9; 3,1 Hz, 8H), 6,97 (ddt, J = 7,3; 4,7; 2,5 Hz, 2H), 6,81 (dt, J = 8,1; 2,5 Hz, 1H), 6,42 (dt, J = 15,9; 2,6 Hz, 1H), 5,95 (t, J = 2,5 Hz, 1H), 4,40 – 4,19 (m, 2H), 4,11 – 3,98 (m, 1H), 3,94 – 3,78 (m, 3H), 3,64 – 3,45 (m, 6H), 3,30 (dt, J = 3,4; 1,9 Hz, 2H), 2,97 (dddd, J = 16,8; 11,6; 8,3; 3,0 Hz, 2H), 2,89 – 2,63 (m, 2H), 2,27 – 2,08 (m, 2H), 2,05 – 1,90 (m, 1H), 1,19 (dd, J = 4,6; 2,6 Hz, 2H). Antiproliferative activity Table. The cell growth inhibitory effects of compounds (6a, 6b) and curcumin evaluated 48 hours after treatment with 3- (4,5) -dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT assay).

Data are expressed as the concentrations which inhibit 50% (IC₅₀) cell growth and are means ± SE of at least three separate experiments. Substance 6b shows selective cytotoxicity to breast cancer cells and colorectal cancer cells (13.3- fold and 50.3-fold, respectively) higher than to normal cells, thus outperforming curcumin, which has these values (4 and 5.9-fold, respectively). ) higher cytotoxicity than in normal cells.

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Claims

1. A compound (6) with the general formula:

where R₁=3: H, OH, OCH₃, N(CH₃)₂ in combination with R₂=3: H, OH, OCH₃, N(CH₃)₂ R₁=4: H, OH, OCH₃, N(CH₃)₂ in combination with R₂=4: H, OH, OCH₃, N(CH₃)₂ R₁=3,4: (OCH₃, OH), (OH, OH), (OCH₃, OTHP), (OCH₃, O(CH₂)_nN(CH₂CH₃)₂ in combination with R₂=3,4: (OCH₃, OH), (OH, OH), (OCH₃, OTHP), (OCH₃, O(CH₂)_nN(CH₂CH₃)₂, optical isomers, polymorphs and pharmaceutically acceptable addition salts, hydrates and solvates thereof.

2. The compound according to the Claim 1, wherein the compound is selected from: (E) -4-phenyl-7-styryl-3,4-dihydro-2H-pyrano [4,

3-b] pyridine-2,5 (1H) -dione (6a) and (E) -7- (4 -hydroxy-3-methoxystyryl) -4-phenyl-3,

4-dihydro-2H-pyrano [4,3-b] pyridine-2,5 (1H) -dione (6b). 3. The compound according to the Claim 2, wherein the structure of the compound is:

4. The compound according to the Claim 2, wherein the structure of the compound is:

5. The compound according to the Claim 1 or 2 for use as a medicament for the treatment of breast and colorectal cancer.

6. A pharmaceutical composition comprising as active ingredient a compound according to the Claim 1 and a pharmaceutically acceptable carrier.

7. A process for the synthesis of a compound (6) according to the Claim 1,

where R₁=3: H, OH, OCH₃, N(CH₃)₂ in combination with R₂=3: H, OH, OCH₃, N(CH₃)₂ R₁=4: H, OH, OCH₃, N(CH₃)₂ in combination with R₂=4: H, OH, OCH₃, N(CH₃)₂ R₁=3,4: (OCH₃, OH), (OH, OH), (OCH₃, OTHP), (OCH₃, O(CH₂)_nN(CH₂CH₃)₂ in combination with R₂=3,4: (OCH₃, OH), (OH, OH), (OCH₃, OTHP), (OCH₃, O(CH₂)_nN(CH₂CH₃)₂ involves the reaction of 4-amino-6-methyl-2H-pyran-2-one (1) with a substituted benzaldehyde (2) and Meldrum's acid (3) in a boiling acid-catalyzed solvent to give DHPDO pyranone derivatives (4) which further is reacted with substituted benzaldehydes (5) in a dry solvent with a base.