Cancer cell proliferation inhibitor composition and processed food

Abstract

To provide a phytol derivative that is contained in a watermelon sprout and has higher cancer cell proliferation inhibitory action than phytol having cancer cell proliferation inhibitory effect, and a component for inhibiting cancer cell proliferation having the derivative as an active ingredient.SOLUTION: A pharmaceutical composition for inhibiting cancer cell proliferation has a substance having a structure expressed by the following formula (1) or (2) as a main component. Because the substance does not affect a normal cell similar to phytol, but has higher cancer cell proliferation inhibitory effect by one digit or more compared to phytol, a pharmaceutical composition containing the substance is hopeful as an anticancer agent with less adverse effect.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a pharmaceutical composition and a processed food which contain a phytol-based substance and have an effect of inhibiting the proliferation of cancer cells. [Background technology]

[0002] It is known that watermelon sprout extracts have the effect of inhibiting the proliferation of cancer cells (Patent Document 1). This document shows that phytol and lutein in watermelon sprout extracts are particularly effective in this regard. Since these substances do not affect normal cells, it is expected that an anti-cancer medical composition with few side effects can be provided. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] International Publication No. 2017 / 131175 Summary of the Invention [Problem to be solved by the invention]

[0004] Phytol and lutein are expected to be used as anti-cancer medical compositions, but the problem is that they need to be administered in large amounts to be effective. [Means for solving the problem]

[0005] The present invention has been conceived in view of the above problems, and provides a phytol-based substance that has no effect on normal cells and further has a high effect of inhibiting the proliferation of cancer cells.

[0006] More specifically, the cancer cell proliferation inhibitor composition of the present invention is characterized by having as an active ingredient at least one of compounds having a structure represented by formula (1) to formula (6) or a pharma- ceutical acceptable salt thereof.

[0007] [ka]

[0008] [ka]

[0009] [ka]

[0010] [ka]

[0011] [ka]

[0012] [ka]

[0013] The present invention can also be provided as a processed food. More specifically, the processed food according to the present invention is characterized in that it contains at least one of the compounds having the structures represented by formulas (1) to (6) or pharma- ceutically acceptable salts thereof.

[0014] In addition, "rac-PT21," "rac-PT22," "rac-PT23," "rac-PT26," "rac-PT27," and "rac-PT28" in formulas (1) to (6) are tentative names of the respective compounds in this specification and are not included in the structure of each compound itself. Effect of the Invention

[0015] The cancer cell proliferation inhibitor composition according to the present invention is expected to be able to inhibit the proliferation of cancer cells without side effects. Therefore, it can be suitably used as an anticancer pharmaceutical composition. In addition, the processed food according to the present invention contains a compound having an inhibitory effect on cancer cell proliferation, so that it is expected to prevent cancer by regularly taking it in the form of a supplement. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] The cancer cell proliferation inhibitor composition and processed food according to the present invention will be described below with reference to examples. Note that the following description is merely an example of one embodiment of the present invention and an example, and the present invention is not limited to the following description. The following description can be modified within the scope of the present invention.

[0017] The cancer cell proliferation inhibitor composition according to the present invention comprises at least one of the phytole compounds represented by the formulae (1) to (6) or pharma- ceutical acceptable salts as an active ingredient.

[0018] [ka]

[0019] [ka]

[0020] [ka]

[0021] [ka]

[0022] [ka]

[0023] [ka]

[0024] The compound (rac-PT21) of formula (1) is (7RS,11RS,E)-1-N,N-dimethylamino-3,7,11,15-tetramethyl-2-hexadecene (dimethylaminophytol) (referred to as (7RS,11RS,E)-1-N,N-dimethylamino-3,7,11,15-tetramethyl-2-hexadecene).

[0025] The compound (rac-PT22) of formula (2) is (7RS,11RS,E)-1-isopropylamino-3,7,11,15-tetramethyl-2-hexadecene (referred to as (7RS,11RS,E)-1-isopropylamino-3,7,11,15-tetramethyl-2-hexadecene).

[0026] The compound (rac-PT23) of formula (3) is (7RS,11RS,E)-1-t-butylamino-3,7,11,15-tetramethyl-2-hexadecene (t-butylaminophytol) (referred to as (7RS,11RS,E)-1-t-butylamino-3,7,11,15-tetramethyl-2-hexadecene).

[0027] The compound (rac-PT26) of formula (4) is (7RS,11RS,E)-N,N,N-trimethyl-3,7,11,15-tetramethyl-2-hexadecenaminium bromide (referred to as (7RS,11RS,E)-N,N,N-trimethyl-3,7,11,15-tetramethyl-2-hexadecenaminium bromide).

[0028] The compound (rac-PT27) of formula (5) is (7RS,11RS,E)-N,N,N-triethyl-3,7,11,15-tetramethyl-2-hexadecenaminium bromide (referred to as (7RS,11RS,E)-N,N,N-triethyl-3,7,11,15-tetramethyl-2-hexadecenaminium bromide).

[0029] The compound of formula (6) (rac-PT28) is 6-amino-1-((7R,11R,E)-3,7,11,15-tetramethyl-2-hexadecenyl)pyrimidin-2(1H)-one (cytosine phytol) (referred to as 6-amino-1-((7R,11R,E)-3,7,11,15-tetramethyl-2-hexadecenyl)pyrimidin-2(1H)-one).

[0030] When these compounds are used as a cancer cell proliferation inhibitor composition (pharmaceutical composition), they may be used alone or as a salt by mixing with a pharmaceutical acceptable acid in a solvent such as water, methanol, ethanol, acetone, etc. Here, examples of the pharmaceutical acceptable acid include inorganic acids such as hydrochloric acid, hydrobromic acid, sulfate, phosphoric acid, nitric acid, etc., 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, ascorbic acid, etc.

[0031] The administration form of the pharmaceutical composition of the present invention is not particularly limited, and may be either oral or parenteral.In addition, it can be made into a suitable dosage form according to the administration form, and can be prepared into various preparations such as injections, or oral preparations such as capsules, tablets, granules, powders, pills, and fine granules, rectal preparations, oily suppositories, and aqueous suppositories.

[0032] The cancer cell proliferation inhibitor composition according to the present invention can also be provided as a processed food, which includes not only general processed foods including luxury foods and health foods such as candy, gum, jelly, biscuits, cookies, rice crackers, bread, noodles, fish and meat paste products, tea, soft drinks, coffee drinks, milk drinks, whey drinks, lactic acid bacteria drinks, yogurt, ice cream, pudding, etc., but also health functional foods such as foods for specified health uses and foods with nutritional functions as specified by the Health Functional Foods System of the Ministry of Health, Labor and Welfare, and further includes nutritional supplements, feed, food additives, etc.

[0033] The processed foods according to the present invention can be prepared by adding the cancer cell proliferation inhibitor composition to the raw materials of these processed foods. EXAMPLES

[0034] <1. Synthesis of Compounds> The synthesis of the compounds of the formulae (1) to (6) will be described below.

[0035] <1-1> di (7RS,11RS,E)-1-bromo-3,7,11,15-tetramethyl-2-hexadecene: Synthesis of (7RS,11RS,E)-1-bromo-3,7,11,15-tetramethyl-2-hexadecene.

[0036] [ka]

[0037] First, a compound of formula (7) was synthesized. "di" is a tentative name for the compound of formula (7) in this specification, and is not included in the structure of the compound of formula (7) itself.

[0038] Add Et2O (diethyl ether) (5 mL) to a 100 mL two-neck flask, add phytol (CAS number: 7541-49-3) (500 mg, 1.686 mmol) dissolved in Et2O (20 mL), and stir for a while at 0°C. Then, add phosphorus tribromide (0.064 mL, 0.674 mmol, 0.4 eq) dropwise and stir at the same temperature for 30 minutes.

[0039] The reaction was confirmed by TLC (Hexane / AcOEt=5 / 1). After confirming that the raw material had disappeared on TLC, the reaction was stopped by dropping a saturated aqueous solution of sodium bicarbonate, extracted with ethyl acetate, and the organic layer was collected and neutralized with a saturated aqueous solution of sodium bicarbonate, washed with brine, dried with anhydrous sodium sulfate, and naturally filtered to remove the sodium sulfate. The solution was concentrated under reduced pressure with a rotary evaporator to obtain 449.9 mg (74%) of a crude product in the form of a reddish brown oil. No purification operation was performed.

[0040] The NMR spectrum of the product obtained by the above procedure is shown below. Yield (74%: reddish brown oil) 1 H NMR (400 MHz, CDCl3): δ 5.53 (1H,t, J = 8.4 Hz), 4.04 (2H,d, J = 8.4 Hz), 2.02 (2H,t, J =7.6 Hz), 1.62-1.50 (2H,m), 1.72 (3H,d, J = 1.3Hz), 1.41-1.02 (21,m), 0.89-0.82(14H,m) and above. From the above results, it was found that the product has the structure of formula (7).

[0041] <1-2> PT21 Synthesis of (7RS,11RS,E)-1-N,N-dimethylamino-3,7,11,15-tetramethyl-2-hexadecene (PT21) In a 5mL screw tube, "di" (24.9mg, 0.07mmol) of formula (7) dissolved in CH2Cl2 (1.0mL) was added and stirred at room temperature. Then, 50% dimethylamine (7μL, 0.14mmol, 2.0eq) was added dropwise and stirred at room temperature for 30 minutes. The end of the reaction was confirmed by TLC (Hexane / AcOEt=5 / 1), and the mixture was diluted with water. The reaction solution was extracted three times with CH2Cl2 and washed three times with saturated saline solution. Then, it was dried with anhydrous sodium sulfate, concentrated under reduced pressure with a rotary evaporator, and dried in vacuum. The obtained product was separated and purified by silica gel open column chromatography (AcOEt (Et3N: triethylamine 1%)), and 9.1mg (40%) of yellow oily product PT21 was obtained.

[0042] The NMR spectrum of the product PT21 is shown below. Yield (26%: Yellow oil). IR (KBr): 1 H NMR (CDCl3, 500 MHz): δ = 5.24 (1H, td, J = 7.0,1.2 Hz), 2.88 (2H, d, J = 7.0 Hz), 2.22 (6H, s), 1.99 (2H, t, J =6.9 Hz), 1.63 (3H, s), 1.57-1.47 (1H, m), 1.47-1.32 (5H, m), 1.32-1.19 (8H, m), 1.19-1.10 (3H, m), 1.10-0.98 (4H, m), 0.90-0.80 (12H, m). 13 C NMR (CDCl3, 125 MHz): δ = 138.8, 121.4, 57.045.2,40.1, 39.4, 37.4, 37.3, 37.2, 36.6, 32.8, 32.7, 27.9, 25.2, 24.8, 24.4, 22.7,22.6, 19.8, 19.7, 16.2. HRMS m / z: [M + H]+ Calcd for C22H46N 324.3630, found324.3636. From the above results, it was found that the product PT21 has the structure of formula (1).

[0043] <1-3> PT22 Synthesis of (7RS,11RS,E)-1-isopropylamino-3,7,11,15-tetramethyl-2-hexadecene (PT22) Isopropylamine (14.0μL, 9.87mg, 0.17mmol, 2.0eq) dissolved in CHCl (0.4mL) was added to a 5mL screw tube and stirred at room temperature. Then, “di” (30.0mg, 0.083mmol) of formula (7) dissolved in CHCl (0.6mL) was added dropwise over 10 minutes and stirred at room temperature for 90 minutes. The reaction was confirmed to be complete by TLC (Hexane / AcOEt=5 / 1), and the mixture was diluted with water. The reaction solution was extracted three times with CHCl and washed three times with saturated saline solution. It was then dried with anhydrous sodium sulfate, concentrated under reduced pressure with a rotary evaporator, and dried in vacuum. The obtained product was separated and purified by silica gel open column chromatography (Hexane / AcOEt=3 / 1 and AcOEt (EtN: triethylamine 1%), and 11.1mg (40%) of yellow oily product PT22 was obtained.

[0044] The NMR spectrum of the product PT22 is shown below. Yield (40%: Yellow oil). IR (KBr): 1H NMR (CDCl3, 500 MHz): δ = 5.25 (1H, td, J =6.8, 1.2 Hz), 3.21 (2H, d, J = 6.7 Hz), 2.82 (1H, m), 1.96 (2H, br m),1.63 (3H, s), 1.57-1.46 (1H, m), 1.46-1.31 (5H, m), 1.31-1.16 (9H, m), 1.16-1.11 (2H, m), 1.06 (10H, br, d, J = 6.3 Hz), 0.89- 0.81 (12H, m). 13C NMR (CDCl3, 125 MHz): δ = 137.6, 122.8, 48.2,44.9, 39.9, 37.4, 37.3, 37.3, 37.2, 36.6, 32.7, 32.6, 27.9, 25.1, 24.7, 24.4,22.9, 22.6, 19.8, 19.7, 16.1. HRMS m / z: [M + H]+ Calcd for C23H48N 338.3787, found338.3785. From the above results, it was found that the product PT22 has the structure of formula (2).

[0045] <1-4> PT23 Synthesis of (7RS,11RS,E)-1-t-butylamino-3,7,11,15-tetramethyl-2-hexadecene (PT23) t-Butylamine (18.0μL, 12.2mg, 0.16mmol, 2.0eq) dissolved in CHCl (2.0mL) was added to a 5mL screw tube and stirred at room temperature. Then, “di” (30.0mg, 0.08mmol) of formula (7) dissolved in CHCl (2.0mL) was added dropwise over 10 minutes and stirred at room temperature for 2.5 hours. The reaction was confirmed to be complete by TLC (Hexane / AcOEt=5 / 1), and the mixture was diluted with water. The reaction solution was extracted three times with CHCl and washed three times with saturated saline solution. It was then dried with anhydrous sodium sulfate, concentrated under reduced pressure with a rotary evaporator, and dried in vacuum. The obtained product was separated and purified by silica gel open column chromatography (Hexane / AcOEt=3 / 1 and AcOEt (triethylamine 1%)), and 13.7mg (49%) of yellow oily product PT23 was obtained.

[0046] The NMR spectrum of PT23 is shown below. Yield (49%: Yellow oil). IR (KBr): 1H NMR (CDCl3, 500 MHz): δ = 5.26 (1H, td, J =6.8, 1.2 Hz), 3.18 (2H, d, J = 6.8 Hz), 1.95 (2H, br, m), 1.63 (3H, s),1.57- 1.47 (1H, m), 1.47-1.32(6H, m), 1.32-1.19 (10H, m), 1.19-1.10 (12H, m), 1.10-0.98 (5H, m), 0.90-0.80 (13H, m). 13C NMR (CDCl3, 125 MHz): δ = 137.7, 123.0, 50.4,40.1, 39.9, 39.3, 37.4, 37.3, 37.3, 36.7, 32.8, 32.7, 28.9, 27.9, 25.2, 24.8,24.4, 22.7, 22.6, 19.7, 19.7, 16.1. HRMS m / z: [M + H]+ Calcd for C24H50N 352.3943, found352.3946. From the above results, it was found that the product PT23 has the structure of formula (3).

[0047] <1-5> PT26 Synthesis of (7RS,11RS,E)-N,N,N-trimethyl-3,7,11,15-tetramethyl-2-hexadecenaminium bromide (PT26) "di" (70.0 mg, 0.2 mmol) of formula (7) dissolved in THF (2.1 mL) was added to a 5 mL screw tube and stirred at room temperature. Then, Me3N (26.0 μL, 0.3 mmol, 1.5 eq) was added dropwise and stirred at room temperature for 23 hours. The progress of the reaction was confirmed by TLC (Hexane / AcOEt = 5 / 1), and Me3N (: trimethyltriamine, 26.0 μL, 0.3 mmol, 1.5 eq) was added. Then, the mixture was stirred for 4 hours (total 27 hours). The completion of the reaction was confirmed by TLC (Hexane / AcOEt = 5 / 1). The reaction solution was concentrated under reduced pressure using a rotary evaporator, AcOEt was added, and 37.7 mg (45%) of white solid PT26 was obtained by suction filtration.

[0048] The NMR spectrum of PT26 is shown below. Yield (45%: white solid). Mp: 199-201℃. IR (KBr): 1H NMR (CDCl3, 400 MHz): δ = 5.34 (1H, t, J =7.8 Hz), 4.27 (2H, d, J = 8.2 Hz), 3.41 (9H, s), 2.12 (2H, br, m), 1.90(3H, s), 1.76 (1H, s), 1.58-1.48 (1H, m), 1.48-1.33 (4H, m), 1.33-1.19 (7H, m), 1.19-0.99 (7H, m), 0.91-0.80 (12H, m). 13C NMR (CDCl3, 100 MHz): δ = 153.6, 110.0, 63.9,52.4, 40.5, 39.3, 37.4, 37.3, 37.2, 36.7, 32.7, 32.6, 27.9, 25.2, 24.7, 24.4,22.7, 22.6, 19.7, 19.6, 17.7. HRMS m / z: [M + H]+ Calcd for C23H48N 338.3787, found338.3787. From the above results, it was found that the product PT26 has the structure of formula (4).

[0049] <1-6> PT27 Synthesis of (7RS11RS,E)-N,N,N-triethyl-3,7,11,15-tetramethyl-2-hexadecenaminium bromide (PT27) In a 5mL screw tube, "di" (50.0mg, 0.14mmol) of formula (7) dissolved in THF (1.5mL) was added and stirred at room temperature. Then, Et3N (60.0μL, 0.42mmol, 3.0eq) was added dropwise and stirred at room temperature for 7 hours, and the completion of the reaction was confirmed by TLC (Hexane / AcOEt=5 / 1). Et2O was added to the reaction solution, and 32.0mg (50%) of white solid PT27 was obtained by suction filtration.

[0050] The NMR spectrum of PT27 is shown below. Yield (50%: white solid). Mp: 132-135℃. IR (KBr): 1H NMR (CDCl3, 400 MHz): δ = 5.19 (1H, td, J = 7.7,1.0 Hz), 4.08 (2H, d, J = 7.7 Hz), 3.47 (6H, q, J = 7.3 Hz), 2.10(2H, br, m), 1.85 (3H, s), 1.66 (2H, s), 1.58-1.48 (1H,m), 1.43 (11H, br, t, J = 7.3 Hz), 1.39-1.31(3H, m), 1.31-1.17 (7H, m), 1.17-1.11 (2H, m), 1.11-0.98 (4H, m), 0.91-0.80 (12H, m). 13C NMR (CDCl3, 100 MHz): δ = 151.8, 109.3, 56.2,53.1, 40.6, 39.3, 37.4, 37.3, 37.2, 36.7, 32.8, 32.6, 27.9, 25.2, 24.8, 24.4,22.7, 22.6, 19.7, 19.6, 17.5, 8.3. HRMS m / z: [M + H]+ Calcd for C26H54N 380.4256, found380.4249. From the above results, it was found that the product PT27 has the structure of formula (5).

[0051] <1-7> PT28 Synthesis of 6-amino-1-((7R,11R,E)-3,7,11,15-tetramethyl-2-hexadecenyl)pyrimidin-2(1H)-one (PT28) Cytosine (184.4 mg, 1.66 mmol, 2.0 eq) dissolved in DMSO (10.0 mL) was added to a 50 mL screw tube and stirred at room temperature. Then, "di" (297.5 mg, 0.83 mmol) of formula (7) dissolved in DMSO (15.0 mL) was added dropwise over 15 minutes and stirred at room temperature for 1 hour. The reaction was extracted with Hexane / AcOEt = 4 / 1, and the completion of the reaction was confirmed by TLC (Hexane / AcOEt = 5 / 1), and the reaction solution was extracted twice with AcOEt and H2O. After further washing with H2O, it was dried with anhydrous sodium sulfate, and the reaction solution was concentrated under reduced pressure using a rotary evaporator and dried in vacuum. The obtained product was separated and purified by silica gel open column chromatography (AcOEt / MeOH (methanol) = 5 / 1), and 59.5 mg (18%) of pale yellow solid PT28 was obtained.

[0052] The NMR spectrum of PT28 is shown below. Yield (18%: pale yellow solid). Mp: 101-102℃. IR (KBr): 3348, 2955, 2928, 1639, 1578, 1547,1462, 1381, 1327, 1292, 1150, 1072, 868, 791, 567 cm-1. 1H NMR (CDCl3, 500 MHz): δ 7.11 (1H, s), 5.66 (1H,d, J = 7.1 Hz), 5.20 (1H, td, J = 6.3, 0.9 Hz), 4.66 (2H, d, J= 6.3 Hz), 2.03-1.98 (2H, m), 1.78 (3H, s), 1.56-1.49 (1H, m), 1.47-1.33 (5H, m), 1.32-1.18 (8H, m), 1.17-1.10 (3H, m), 1.10-0.99 (4H, m), 0.91-0.76 (13H, m). 13C NMR (CDCl3, 125 MHz): δ 158.2, 153.0, 141.1,117.5, 98.2, 41.0, 40.3, 39.9, 39.4, 37.4, 37.4, 37.3, 36.8, 32.8, 32.7, 28.0,25.2, 24.8, 24.5, 22.8, 22.7, 19.8, 19.7, 16.6. HRMS m / z: [M + Na]+ Calcd for C24H43N3ONa 412.3304; Found 412.3306. From the above results, it was found that the product PT28 has the structure of formula (6).

[0053] <2. Evaluation of the inhibitory activity of compounds on the proliferation of human leukemia T cell line Jurkat cells> The cancer cell proliferation inhibitory activity of each of the above samples was examined using human leukemia T cell line Jurkat cells.

[0054] 1×10 5 50μL of Jurkat cell suspension adjusted to 100 cells / mL was seeded into wells of a 96-well plate. The DMSO solution of the evaluation sample was diluted 500-fold with culture medium. Immediately after that, 50μL of the diluted sample solution was added to start cell treatment. After stirring for 30 seconds with a microplate reader, the cells were cultured in an incubator. After 72 hours, 10μL of the live cell measurement reagent SF was added. After a color reaction in the incubator for 2-3 hours, the absorbance at 450nm and 630nm was measured with a microplate reader, and the cell viability was calculated using the following formula (F1). Cell viability (%)=[(As-Ab) / (Ac-Ab)]×100...(F1) In addition, As, Ab, and Ac are as follows.

[0055] As: absorbance of evaluation well Ab: absorbance of negative control wells Ac: absorbance of control wells The "absorbance" in the above formula is the absorbance at 450 nm (A 450) to the absorbance at 630 nm (A 630 ) is subtracted from "A 450 -A 630 In addition, a linear approximation was performed on the logarithmic graph of the two concentrations that narrowly sandwiched the cell viability of 50%, and the IC 50 The results are shown in Table 1.

[0056] [Table 1]

[0057] Table 1 shows the structural formula and IC of each sample. 50 The values ​​(μM) are shown. As references, phytol (rac-PT0) and aminophytol (rac-PT10) ((7R,11R,E)-1-amino-3,7,11,15-tetramethyl-2-hexadecene) were also measured simultaneously.

[0058] As a result, the concentrations of phytol (rac-PT0) and aminophytol (rac-PT10) were 29.8 μM and 3.8 μM, respectively.

[0059] On the other hand, cytosine phytol (PT28) was 3.2 μM, and t-butylaminophytol (PT23) was 3.1 μM.

[0060] Furthermore, dimethylaminophytol (PT21) was 2.8 μM, and isopropylaminophytol (PT22) was 2.6 μM.

[0061] In addition, trimethylphytol ammonium bromide (PT26) was 1.41 μM, and triethylphytol ammonium bromide (PT27) was 0.48 μM.

[0062] In other words, it was found that these six compounds according to the present invention are more effective than phytol by one to two orders of magnitude. [Industrial Applicability]

[0063] The cancer cell proliferation-inhibiting composition according to the present invention can be suitably used for the treatment and prevention of cancer.

Claims

1. A composition for inhibiting the proliferation of cancer cells, comprising as an active ingredient at least one of compounds having a structure represented by any one of formulas (1) to (6) or a pharma- ceutical acceptable salt thereof. 【Chemistry 100】 【Chemistry 101】 【Chemistry 102】 【Chemistry 103】 【Chemistry 104】 【Chemistry 105】

2. A processed food comprising at least one compound having a structure represented by any one of formulas (1) to (6) or a pharma- ceutically acceptable salt thereof. 【Chemistry 106】 【Chemistry 107】 【Chemistry 108】 【Chemistry 109】 【Chemistry 110】 【Chemistry 111】

Images