Cancer cell proliferation inhibiting composition and processed food

Abstract

It is known that phytol contained in watermelon seedlings has a cancer cell proliferation inhibitory effect. However, the problem is that a large amount of phytol needs to be taken in order to exhibit the cancer cell proliferation inhibitory effect. A cancer cell proliferation inhibitory composition having at least one of a compound having a structure represented by Formula (1), Formula (2), Formula (6), Formula (7), or Formula (8) or a pharmaceutically acceptable salt thereof as a main component has a higher cancer cell proliferation inhibitory effect than phytol.

Description

Technical Field

[0001] This invention relates to pharmaceutical compositions and processed foods containing chlorophyll-like substances and having an inhibitory effect on cancer cell proliferation. Background Technology

[0002] It is known that extracts from watermelon seedlings have an inhibitory effect on cancer cell proliferation (Patent Document 1). This document shows that chlorophyll and lutein, in particular, are effective in achieving this effect in watermelon seedling extracts. Furthermore, since these substances do not affect normal cells, it is expected that they can provide anticancer drug compositions with fewer side effects.

[0003] Existing technical documents

[0004] Patent documents

[0005] Patent Document 1: International Publication No. 2017 / 131175 Summary of the Invention

[0006] The technical problem that the invention aims to solve

[0007] Phloxiol and lutein are expected to be used in anticancer drug combinations. However, the problem is that relatively large quantities are required to achieve the desired effect.

[0008] Technical solutions adopted to solve technical problems

[0009] Based on the above-mentioned problems, the present invention provides a substance with a high inhibitory effect on cancer cell proliferation.

[0010] More specifically, the cancer cell proliferation inhibitory composition of the present invention is characterized in that it uses at least one of a compound having a structure represented by formula (1), formula (2), formula (6), formula (7) or formula (8) or a pharmaceutically permissible salt thereof as a main component.

[0011] [Chemical Formula 1]

[0012]

[0013] [Chemical Formula 2]

[0014]

[0015] [Chemical Formula 3]

[0016]

[0017] [Chemical Formula 4]

[0018]

[0019] [Chemical Formula 5]

[0020]

[0021] In addition, “rac-1”, “rac-2”, “rac-6”, “rac-7” and “rac-8” in formulas (1), (2), (6), (7) and (8) are temporary names of the compounds in this specification and are not included in the structure of the compounds themselves.

[0022] Furthermore, the present invention can also be provided as a processed food. More specifically, the processed food of the present invention is characterized in that it comprises a compound having the structure represented by formula (1), formula (2), formula (6), formula (7) or formula (8) or a pharmaceutically permissible salt thereof. In addition, the compounds having the structures of formula (2), formula (6), formula (7), and formula (8) are all synthesized for the first time and are new substances.

[0023] Invention Effects

[0024] The cancer cell proliferation-inhibiting composition of the present invention can inhibit the proliferation of cancer cells. Therefore, it can be advantageously used as an anticancer pharmaceutical composition. Furthermore, since the processed food of the present invention contains compounds with cancer cell proliferation-inhibiting effects, cancer prevention can be achieved through regular consumption as a supplement. Attached Figure Description

[0025] Figure 1 The image shows the inhibitory effect of phytol on cancer cell proliferation.

[0026] Figure 2 The image shows the inhibitory effect of chlorophyll on cancer cell proliferation.

[0027] Figure 3 The image shows the inhibitory effect of chlorophyllamide on cancer cell proliferation.

[0028] Figure 4 The image shows the inhibitory effect of (mono)methyl chlorophyllamine on cancer cell proliferation.

[0029] Figure 5 The image shows the inhibitory effect of (monohexyl chlorophyll) on cancer cell proliferation.

[0030] Figure 6 The image shows the inhibitory effect of acetylated chlorophyllamine on cancer cell proliferation. Detailed Implementation

[0031] The following description, including accompanying drawings and embodiments, illustrates the cancer cell proliferation inhibitory composition and processed food products related to the present invention. Furthermore, the following description only illustrates one embodiment and one example of the invention, and the invention is not limited to the following description. Changes may be made to the following description without departing from the spirit of the invention.

[0032] The cancer cell proliferation inhibitory composition of the present invention comprises a compound having a structure represented by formula (1), formula (2), formula (6), formula (7) or formula (8) or a pharmaceutically permissible salt thereof.

[0033] [Chemical Formula 6]

[0034]

[0035] [Chemical Formula 7]

[0036]

[0037] [Chemical Formula 8]

[0038]

[0039] [Chemical Formula 9]

[0040]

[0041] [Chemical Formula 10]

[0042]

[0043] Compound (rac-1) of formula (1) is (E)-3,7,11,15-tetramethylhexadec-2-en-1-amine (hereinafter referred to as "chlorophyllamine"), and compound (rac-2) of formula (2) is (E)-3,7,11,15-tetramethylhexadec-2-enamide (hereinafter referred to as "chlorophyllamide").

[0044] Compound (rac-6) of formula (6) is (E)-N,3,7,11,15-pentamethylhexadec-2-en-1-amine (hereinafter referred to as "methyl chlorophyllamine" or "monomethyl chlorophyllamine"), and compound (rac-7) of formula (7) is (E)-N-hexyl-3,7,11,15-tetramethylhexadec-2-en-1-amine (hereinafter referred to as "methyl chlorophyllamine" or "monomethyl chlorophyllamine"). 1,15-tetramethylhexadec-2-en-1-amine (hereinafter referred to as "hexyl chlorophyllamine" or "monohexyl chlorophyllamine"), compound (rac-8) of formula (8) is (E)-N-(3,7,11,15-tetramethylhexadec-2-en-1-yl)acetamide (hereinafter also referred to as "acetylated chlorophyllamine").

[0045] When these compounds are used as cancer cell proliferation inhibitory compositions (pharmaceutical compositions), in addition to being used alone, they can also be used as salts by mixing with pharmaceutically permissible acids in solvents such as water, methanol, ethanol, and acetone. Examples of pharmaceutically permissible acids include inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, and nitric acid, and organic acids such as acetic acid, propionic acid, oxalic acid, succinic acid, lactic acid, malic acid, tartaric acid, citric acid, maleic acid, fumaric acid, methanesulfonic acid, p-toluenesulfonic acid, and ascorbic acid.

[0046] The administration method of the pharmaceutical composition of the present invention is not particularly limited and can be any oral or non-oral administration method. Furthermore, it can be formulated into appropriate dosage forms according to the administration method, such as injections, capsules, tablets, granules, powders, pills, fine granules, oral preparations, rectal preparations, oily suppositories, aqueous suppositories, and other formulations.

[0047] Furthermore, the cancer cell proliferation inhibitory composition involved in this invention can also be provided as a processed food. As processed foods, these include not only general processed foods containing snacks or health foods such as sugar, chewing gum, jelly, biscuits, cookies, rice cakes, bread, noodles, processed fish and meat products, tea, soft drinks, coffee drinks, milk drinks, whey drinks, lactic acid bacteria drinks, yogurt, ice cream, and pudding, but also health functional foods such as foods for specific health use or nutritional functional foods as defined by the Health Functional Foods Regulations of the Ministry of Health, Labour and Welfare. They can also be further included in processed foods such as nutritional supplements, animal feed, and food additives.

[0048] The processed foods involved in this invention can be formulated by adding a cancer cell proliferation inhibitory composition to the raw materials of these processed foods.

[0049] Example

[0050] <1. Synthesis of Compounds>

[0051] The following are examples of the synthesis of chlorophyllamine and chlorophyllamide.

[0052] <1-1>

[0053] Synthesis of (E)-1-Bromo-3,7,11,15-tetramethylhexadec-2-ene: (E)-1-bromo-3,7,11,15-tetramethylhexadec-2-ene (rac-3)

[0054] [Chemical Formula 11]

[0055]

[0056] First, the compound of formula (3) is synthesized. “rac-3” is a temporary designation for the compound of formula (3) in this specification, which is not included in the structure of the compound of formula (3).

[0057] Add 5 mL of Et₂O to a 100 mL two-necked flask, then add 500 mg (1.686 mmol) of phytol dissolved in 20 mL of Et₂O, and stir at 0 °C for a period of time. Then, add 0.064 mL (0.674 mmol, 0.4 eq) of phosphorus tribromide dropwise, and stir at the same temperature for 30 minutes.

[0058] The reaction was confirmed by TLC (Thin-Layer Chromatography) (hexane / AcOEt = 5 / 1). After confirming the disappearance of the starting material on the TLC, the reaction was stopped by adding saturated sodium bicarbonate aqueous solution. The mixture was extracted with ethyl acetate, and the organic layer was collected, neutralized with saturated sodium bicarbonate aqueous solution, washed with brine, dried with anhydrous sodium sulfate, and the sodium sulfate was removed by natural filtration. The solution was then concentrated under reduced pressure using a rotary evaporator to obtain 449.9 mg of a reddish-brown oily crude product. No purification was performed.

[0059] Yield (74%: reddish-brown oily substance) 1H NMR (CDCl3, 400MHz): δ0.82-0.89 (m, 14H), 1.02-1.41 (m, 21H), 1.50-1.62 (m, 2H), 1.7 2 (d, 3H, J=1.3Hz), 2.02 (t, 2H, J=7.6Hz), 4.04 (d, 2H, J=8.4Hz), 5.53 (t, 1H, J=8.4Hz).

[0060] <1-2>

[0061] Synthesis of (E)-(3,7,11,15-tetramethylhexadec-2-en-1-yl)isoindoline-1,3-dione: (E)-(3,7,11,15-tetramethylhexadec-2-en-1-yl)isoindoline-1,3-dione (rac-4)

[0062] [Chemical Formula 12]

[0063]

[0064] Subsequently, compound (4) was synthesized using rac-3 as a starting material. “rac-4” is a provisional designation for the compound with structure (4) in this specification, and it is not included in the structure of the compound with structure (4).

[0065] Under a nitrogen atmosphere, rac-3 (449.9 mg, 1.252 mmol) dissolved in DMF (7 mL) was added to a 100 mL three-necked flask, and the mixture was stirred at room temperature for a period of time. Then, potassium phthalimide (301.4 mg, 1.627 mmol, 1.3 eq) dissolved in DMF (5 mL) was added, and the mixture was stirred at the same temperature for 2 hours.

[0066] The reaction was confirmed by TLC (hexane only). After confirming the disappearance of the starting material on the TLC, the reaction was stopped by adding H2O dropwise at the same temperature, followed by extraction with ethyl acetate. The organic layer was collected and neutralized with a saturated aqueous sodium bicarbonate solution. After washing with brine, the solution was dried over anhydrous sodium sulfate, and the sodium sulfate was removed by natural filtration. The solution was concentrated under reduced pressure using a rotary evaporator to obtain 591.5 mg of a yellow crystalline crude product.

[0067] The crude product was purified by open column chromatography (hexane / AcOEt = 20 / 1) to obtain 422.3 mg of yellow oily product.

[0068] Yield (79%: yellow oil) 1H NMR (CDCl3, 400MHz): δ0.80-0.88 (m, 12H), 0.99-1.57 (m, 19H), 1.82 (s, 3H), 1.95 (t, J=7.5H z), 4.28 (d, 2H, J=7.3Hz), 5.26 (tq, 1H, J=7.3, 1.2Hz), 7.81-7.86 (m, 2H), 7.67-7.72 (m, 2H).

[0069] <1-3> Synthesis of chlorophyll

[0070] Synthesis of (E)-3,7,11,15-tetramethylhexadec-2-en-1-amine: (E)-3,7,11,15-tetramethylhexadec-2-en-1-amine (chlorophyllamine (rac-1))

[0071] Chloramine (rac-1) was synthesized using rac-4 as a starting material. Under a nitrogen atmosphere, 300 mg (0.705 mmol) of rac-4 dissolved in 10 mL of ethanol was added to a 100 mL three-necked flask, followed by dropwise addition of hydrazine monohydrate (0.103 mL, 2.114 mmol, 3.0 eq). The mixture was stirred at room temperature for 2 hours. The reaction was confirmed by TLC (hexane / AcOEt = 20 / 1).

[0072] After confirming the disappearance of the starting material on TLC, the sample was filtered, the filtrate was recovered and extracted with ethyl acetate, the organic layer was collected and washed with brine, dried with anhydrous sodium sulfate, and the sodium sulfate was removed by natural filtration. The solution was concentrated under reduced pressure using a rotary evaporator to obtain 246.9 mg of a yellow crystalline crude product. The crude product was purified by open column chromatography (MeOH (methanol) / CH2Cl2 (dichloromethane) = 95 / 5, containing 1% 1M ammonia) to obtain 64.5 mg (31%) of a yellow oily product (chlorophyllin).

[0073] Yield (31%): Yellow oily 1 H NMR (CDCl3, 400MHz): δ0.80-0.91 (m, 12H), 1.02-1.55 (m, 21H), 1.61 (s, 3H), 1.96 (t, 2H, J=7.9Hz), 3.28 (d, 2H, J=6.9Hz), 5.25 (tq, 1H, J=6.9, 1.3Hz).

[0074] <1-4> Synthesis of Chloramide

[0075] Synthesis of (E)-3,7,11,15-tetramethylhexadec-2-enamide: (E)-3,7,11,15-tetramethylhexadec-2-enamide (chlorophyllamide (rac-2))

[0076] Phytanic acid (CAS No.: 14721-66-5) was synthesized, and chlorophyllamide was then synthesized from phytanic acid.

[0077] First, phytic acid of formula (5) (rac-5) is synthesized. “rac-5” is a provisional designation for phytic acid of formula (5) in this specification, which is not included in the structure of the compound of formula (5).

[0078] [Chemical Formula 13]

[0079]

[0080] A mixture of acetonitrile (0.150 mL) and water (0.150 mL) was added to a 10 mL two-necked flask. Diacetoxyiodobenzene (477.9 mg, 1.484 mmol, 2.2 eq) and a catalytic amount of TEMPO (21.1 mg, 0.135 mmol, 0.2 eq) were then added dropwise at room temperature under a nitrogen atmosphere. Subsequently, phytol (200 mg, 0.674 mmol) dissolved in a mixture of acetonitrile (0.750 mL) and water (0.750 mL) was added dropwise at the same temperature under a nitrogen atmosphere. The mixture was stirred at room temperature for 14 hours under a nitrogen atmosphere.

[0081] The reaction was confirmed by TLC (hexane / AcOEt = 5 / 1). Although the starting material was still present on the TLC, the reaction could not be confirmed, so it was extracted with ethyl acetate. The organic layer was collected, washed with brine, dried with anhydrous sodium sulfate, and the sodium sulfate was removed by natural filtration. The solution was concentrated under reduced pressure using a rotary evaporator to obtain 308.6 mg of a red oily crude product. The crude product was purified by open column chromatography (hexane / AcOEt = 15 / 1) to obtain 47.5 mg (23%) of a yellow oily product. However, due to insufficient purity, it was further purified by HPLC to obtain 21.8 mg (yield 10%) of a yellow oily phytonic acid (rac-5).

[0082] Yield (23%): Yellow oily 1 H NMR (CDCl3, 400MHz): δ0.80-0.88 (m, 12H), 1.05-1.53 ​​(m, 19H), 2.07-2.025 (m, 5H), 5.69 (bs, 1H). 13C NMR (CDCl3, 100MHz): δ19.1, 19.7, 19.8, 22.6, 22.7, 24.5, 24.8, 24.9, 28.0, 32.6, 32.8, 36.5, 37.3, 37.4, 39.4, 41.5, 115.0, 153.1, 163.6.

[0083] Phytanic acid (rac-5) (96.8 mg, 0.312 mmol) dissolved in THF (1.0 mL) was added to a 5 mL sample vial, followed by dropwise addition of 28% ammonia solution (0.200 mL, 2.959 mmol, 9.5 eq), and the mixture was stirred at room temperature for a period of time. Then, DMT-MM (4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholine chloride n-hydrate: 129.4 mg, 0.468 mmol, 1.5 eq) was added as a condensing agent, and the mixture was stirred at room temperature for 16 hours. The reaction was confirmed by TLC (hexane / AcOEt = 3 / 1).

[0084] Although the starting material was still present on TLC, the reaction could not be confirmed. Therefore, it was extracted with ethyl acetate, washed with brine, dried with anhydrous sodium sulfate, and the sodium sulfate was removed by natural filtration. The solution was then concentrated under reduced pressure using a rotary evaporator to obtain 104.5 mg of white crystalline crude product. The crude product was purified by open column chromatography (hexane / AcOEt = 1 / 1) to obtain 14.1 mg (15%) of white crystalline product. However, due to insufficient purity, it was further purified by HPLC to obtain 7.2 mg (yield 8%) of white crystalline product (chlorophyllamide).

[0085] Yield (15%): White solid 1 H NMR (CDCl3, 400MHz): δ0.80-0.91 (m, 12H), 1.02-1.56 (m, 19H), 2.09 (td, 2H, J=7.7, 2.2Hz), 2.15 (d, 3H, J=1.0Hz), 5.31 (br s, 2H), 5.61 (d, 1H, J=1.2Hz). HRMS m / z[M+Na] + With C 20 H 39 NaNO + Calculated value: 377.2929; Actual value: 377.2929.

[0086] <1-5> Synthesis of Methylphyllin

[0087] Synthesis of (E)-N,3,7,11,15-pentamethylhexadec-2-en-1-amine: (E)-N,3,7,11,15-pentamethylhexadec-2-en-1-amine (rac-6)

[0088] [Chemical Formula 14]

[0089]

[0090] Add 40% methylamine (0.740 mL, 21.0 mmol, 33.4 eq) dissolved in CH₂Cl₂ (4.0 mL) to a 20 mL sample vial and stir at room temperature. Then, add rac-3 (225.0 mg, 0.630 mmol) dissolved in CH₂Cl₂ (4.0 mL) dropwise over 10 minutes, stirring at room temperature for 1 hour. The reaction was confirmed by TLC (hexane / AcOEt = 5 / 1).

[0091] After confirming the disappearance of the starting material on TLC, water was added dropwise for dilution, followed by extraction with CH2Cl2 and washing with brine. The solution was then dried with anhydrous sodium sulfate, and sodium sulfate was removed by natural filtration. The solution was concentrated under reduced pressure using a rotary evaporator to obtain 169.1 mg of a yellow oily crude product. This crude product was purified by open column chromatography (hexane / AcOEt = 1 / 1) to obtain 77.8 mg of a yellow oily product. Due to insufficient purity, it was purified again by open column chromatography (MeOH / AcOEt = 1 / 1, triethylamine 1%) to obtain 28.5 mg (15%) of a yellow oily product. Finally, it was purified again by open column chromatography (MeOH / AcOEt = 1 / 5, triethylamine 1%) to obtain 17.8 mg (9%) of a yellow oily product.

[0092] Yield (9%: yellow solid) 1 H NMR (CDCl3, 500MHz): δ5.24 (1H, td, J=13.5, 1.4Hz), 3.19 (2H, d, J=6.9Hz), 2.42 (2H, s), 2.01-1.88 (3H, m), 1.63 (3H, s), 1.57-1.48 (1H, m), 1.46-1.16 (13H, m), 1.16-0.99 (7H, m), 0.89-0.80 (12H, m). 13C NMR (CDCl3, 100MHz): δ 137.42, 120.95, 47.8, 38.7, 38.1, 36.2, 36.2, 36.1, 35.4, 34.6, 31.6, 31.5, 26.8, 24.0, 23.6, 23.3, 21.5, 21.4, 18.6, 15.0. HRMS m / z[M+H] + With C 21 H 44 N + Calculated value: 310.3468; Actual value: 310.3477.

[0093] <1-6> Synthesis of Hexylchlorophyllin

[0094] Synthesis of (E)-N-hexyl-3,7,11,15-tetramethylhexadec-2-en-1-amine: Synthesis of (E)-N-hexyl-3,7,11,15-tetramethylhexadec-2-en-1-amine (rac-7)

[0095] [Chemical Formula 15]

[0096]

[0097] Add 4.20 mL of n-hexylamine (3.20 g, 31.7 mmol, 33.4 eq) dissolved in 5 mL of CH₂Cl₂ to a 50 mL sample vial and stir at room temperature. Then, add dropwise rac-3 (342.7 mg, 0.950 mmol) dissolved in 5 mL of CH₂Cl₂, continuing for 10 minutes, and stirring at room temperature for 2 hours. The reaction was confirmed by TLC (hexane / AcOEt = 5 / 1).

[0098] After confirming the disappearance of the starting material on TLC, water was added dropwise for dilution, followed by extraction with CH2Cl2 and washing with brine. The solution was then dried with anhydrous sodium sulfate, and sodium sulfate was removed by natural filtration. Water was added to the solution, and reagent residue was removed using a rotary evaporator, yielding 352.5 mg of a yellow oily crude product. This crude product was purified by open column chromatography (hexane / AcOEt = 10 / 1 with AcOEt (triethylamine 1%)), yielding 270.2 mg of a yellow oily product.

[0099] Yield (75%: yellow solid). 1H NMR (CDCl3, 500MHz): δ5.25 (1H, td, J=13.9, 1.3Hz), 3.21 (2H, d, J=3.2Hz), 2.59 (2H, t, J=2.5Hz), 2.01- 1.90 (2H, m), 1.62 (3H, s), 1.57-1.44 (3H, m), 1.44-1.17 (19H, m), 1.17-1.00 (7H, m), 0.91-0.81 (15H, m). 13 C NMR (CDCl3, 125MHz): δ137.8, 122.7, 49.6, 47.3, 39.9, 39.3, 37.4, 37.3, 37.2, 36.6 , 32.7, 32.6, 31.8, 30.1, 27.9, 27.1, 25.1, 24.7, 24.4, 22.7, 22.6, 19.7, 16.1, 14.0. HRMS m / z[M+Na] + With C 26 H 51 NaNO + Calculated value: 402.4070; Actual value: 402.4076.

[0100] <1-7> Synthesis of Acetylchlorophyllin

[0101] (E)-N-(3,7,11,15-tetramethylhexadec-2-en-1-yl)acetamide:

[0102] Synthesis of (E)-N-(3,7,11,15-tetramethylhexadec-2-en-1-yl)acetamide (rac-8)

[0103] [Chemical Formula 16]

[0104]

[0105] Add rac-1 (100 mg, 0.340 mmol) and triethylamine (0.15 mL, 1.1 mmol, 3.1 eq) dissolved in 1.0 mL of dry THF to a 5 mL sample vial, and stir at 0 °C under a nitrogen atmosphere. Then, add acetyl chloride (0.040 mL, 0.53 mmol, 1.5 eq) dissolved in 1.0 mL of dry THF dropwise, and stir at the same temperature for 2 hours under a nitrogen atmosphere. The reaction was confirmed by TLC (hexane / AcOEt = 1 / 1).

[0106] After confirming the disappearance of the starting material on TLC, water was added dropwise to stop the reaction. The mixture was extracted with ethyl acetate, the organic layer was collected and washed with water, dried over anhydrous sodium sulfate, and the sodium sulfate was removed by natural filtration. The solution was concentrated under reduced pressure using a rotary evaporator to obtain 103.1 mg of a yellow oily crude product. The crude product was purified by open column chromatography (hexane / AcOEt = 2 / 1) to obtain 68.5 mg (60%) of a yellow oily product.

[0107] Yield (60%): Yellow solid 1 H NMR (CDCl3, 400MHz): δ5.35 (1H, br s), 5.18 (1H, td, J = 14.0, 1.4Hz), 3.84 (2H, t, J = 12.2Hz), 1.97 (5H, s), 1.65 (4H, s), 1.56-1.47 (1H, m), 1.46-1.18 (1H, m), 1.16-1.00 (7H, m), 0.88-0.82 (13H, m). 13 C NMR (CDCl3, 100MHz): δ169.8, 140.6, 119.5, 39.8, 39.4, 37.6, 37.4, 37.3, 37.2 , 36.7, 32.8, 32.7, 28.0, 25.2, 24.8, 24.5, 23.3, 22.7, 22.6, 19.8, 19.7, 16.2. HRMS m / z[M+Na] + With C 22 H 43 NaNO + Calculated value: 360.3237; Actual value: 360.3235.

[0108] <2. Cell Culture>

[0109] The human leukemia T-cell line Jurkat cells were obtained from the RIKEN Bioresource Center (Tsukuba City, Ibaraki Prefecture). They were cultured in RPMI 1640 medium (Wako Pure Pharmaceutical Industries Co., Ltd., Osaka City, Osaka Prefecture) at 37°C at 95% air - 5% CO2, containing 10% bovine embryonic serum (Thermo Fisher Scientifics, KK, MA, USA), 100 U / mL penicillin, and 100 μg / mL streptomycin (both from Life Technologies, Carlsbad, CA, USA).

[0110] <3. Effects of the compound on the proliferation of human leukemia T cell line Jurkat>

[0111] Adjust Jurkat cells to 1×10 5 Cells were seeded sequentially at 500 μL / well in 24-well multi-well plates (ThermoFisher Scientifics KK). After seeding, phytol, phytamine, phytamide, methylphytamine, hexylphytamine, and acetylphytamine were added, and the concentrations were adjusted with distilled water to a final concentration of 10 μM, 25 μM, and 50 μM, respectively. No compounds were added to the control group. After adding each compound, cells were stained with trypan blue (Life Technologies) at 24, 48, and 72 hours, and viable cells were counted using a hemocytometer.

[0112] The results are as follows Figures 1-6 As shown. The horizontal axis represents the culture time (hours), and the vertical axis represents the number of surviving cells (×10). 5 (cells / mL). In the figure, "Cont" represents the control example, and "10μM", "25μM", and "50μM" represent the concentrations. Figure 1 It is chlorophyll. Figure 2 It is chlorophyll. Figure 3 It is chlorophyllamide. Figure 4 It is monomethyl chlorophyllin. Figure 5 It is monohexyl chlorophyllamine. Figure 6 It is acetylated chlorophyll. As shown in Patent Document 1, chlorophyll can inhibit the proliferation of Jurkat cells (cancer cells) in a concentration-dependent manner.

[0113] On the other hand, chlorophyllamine, chloroamide, monomethylchlorophyllamine, monohexylchlorophyllamine, and acetylated chlorophyllamine showed better effects than chlorophyll. More specifically, for chlorophyllamine, viable cells could still be identified even at a concentration of 50 μM after 72 hours, while for chlorophyllamine, chloroamide, monomethylchlorophyllamine, and monohexylchlorophyllamine, almost no viable cells were found after 24 hours at a concentration of 25 μM. For acetylated chlorophyllamine, viable cells could still be identified at a concentration of 25 μM, but almost no viable cells were found at a concentration of 50 μM.

[0114] The figure shows the IC50 results for each compound. Phytol was 29.8 μM, while chlorophyllamine was 3.8 μM, chlorophyllamide was 9.6 μM, monomethylchlorophyllamine was 6.6 μM, and monohexylchlorophyllamine was 8.5 μM, indicating an effect more than one order of magnitude higher than chlorophyll. Additionally, acetylchlorophyllamine was 21.2 μM, lower than chlorophyll.

[0115] Industrial applications

[0116] The cancer cell proliferation inhibitory composition of the present invention can be advantageously used for the treatment or prevention of cancer.

Claims

1. A leukemia cancer cell proliferation inhibitory composition which takes at least one of a compound having a structure represented by Formula (1), Formula (2), Formula (6), Formula (7), or Formula (8), or a pharmaceutically acceptable salt thereof as a main ingredient: [Chemical Formula 100] [Chemical Formula 101] [Chemical Formula 102] [Chemical Formula 103] [Chemical Formula 104] 2. A compound having a structure of Formula (2): [Chemical Formula 110] 3. A compound having a structure of Formula (7): [Chemical Formula 112] 4. A compound having a structure of Formula (8): [Chemical Formula 113]

Images