KR101599195B1 - Novel carotenoid nanoparticles - Google Patents

Abstract

The present invention relates to nanoparticle morphology using carotenoid compounds having a partially different structure from conventional carotenoid compounds.
The carotenoid nanoparticles comprising the novel carotenoid compound as particles having a diameter of 1000 nm or less according to the present invention are remarkably improved in terms of stability and functionality as compared with nanoparticles prepared from existing beta carotene compounds. Specifically, the carotenoid nanoparticles of the present invention can be safely absorbed in vivo, can be carried to a place where oxidative stress is intact, and can be used for delivery of nutrients, medicines, cosmetics and the like which have poor water absorption and persistence.

Description

Novel carotenoid nanoparticles < RTI ID = 0.0 >

The present invention relates to nanoparticle morphology using carotenoid compounds having a partially different structure from conventional carotenoid compounds.

Carotenoids are red and yellow pigments widely distributed in plants and animals such as carrots, tomatoes, and shrimps. Carotenoids have various physiological activities such as contributing to antioxidant activity in vivo or helping immune function, and most act as provitamin-A.

Almost all animals, including humans, can not synthesize carotenoids themselves, so most of them are fed through food. However, even the carotenoids in these foods are known to have poor conversion to vitamin A in vivo and low bioavailability. In addition, carotenoids contain many double bonds in molecules and are unstable substances that are easily oxidized to air, heat, and light.

Conventionally, there are two solutions to such problems. There are two methods for solving these problems. One is a method of dissolving carotenoid, which is a fat-soluble component, in oil components, and a method of making it into water-like nanoparticles. In the case of carotenoids dissolved in oil, unstable forms of carotenoids promote the acidosis of oil, and this rancorous phenomenon again causes a chain reaction in which carotenoids are decomposed, and the amount of carotenoids that can be included in oil is also limited. There is also a problem that the bioavailability and conversion rate in vivo are not good.

In addition, even if carotenoids are prepared as nanoparticles, the antioxidant ability of the nanoparticles is significantly reduced when the nanoparticles are exposed to sunlight due to the instability of the carotenoids themselves. There has been a problem in that it accompanies a change in morphology such as reduction.

Accordingly, the inventors of the present invention confirmed that, when nanoparticles were prepared using a carotenoid compound having a novel structure to produce carotenoid nanoparticles having the above problems, nanoparticles improved in biological utility and functionality could be provided Thus completing the present invention.

The present invention is to provide carotenoid nanoparticles comprising a novel carotenoid compound as particles having a diameter of 1000 nm or less.

In order to solve the above problems, the present invention provides carotenoid nanoparticles comprising a carotenoid compound represented by the following formula (1) or (2) as particles having a diameter of 1000 nm or less.

[Chemical Formula 1]

(2)

Bis (4-methoxyphenyl) -4,15-dimethyloctadeca- 4-methoxyphenyl-4-methoxyphenyl-4-methoxyphenylcarbamate represented by the above formula (BAS carotenoid, ((1E, 3E, 5E, 7Z, 9E, 11Z, 13E, 15E, 17E) 1,1-diyl) bis (4,1-phenylene)) bis (methylsulfane)) or a carotenoid compound represented by the above formula (2) BTS carotenoid, ((1E, 3E, 5E, 7Z, 9E, 11Z, 13E, 15E, 17E) -4,15-dimethyl-7,12- 11,13,15,17-nonaene-1,18-diyl bis (4,1-phenylene)) bis (methylsulfane)) is a tetraterpene- methoxy group or a methyl group.

The carotenoid compound represented by the formula (1) or (2) of the present invention has a structure in which a methyl (phenyl) sulfanyl group is substituted at both ends in terms of chemical structure, and is structurally more stable.

The carotenoid compound represented by Formula 1 or Formula 2 of the present invention is a particle having a diameter of 1000 nm or less and forms a carotenoid nanoparticle form, more preferably a nanoparticle form having a diameter of 1 nm to 100 nm .

 The nanoparticle form of the present invention can be prepared by a known method for producing nanoparticles known in the art. In one embodiment of the present invention, nanoparticles were prepared by the in-water method.

In addition, the nanoparticles may further comprise a surfactant. The surfactant may be selected from the group consisting of sodium linear alkylbenzenesulfonate, sodium alpha-sulfonate, sodium alpha-olefinsulfonate, sodium alkyl etheresterate, polyoxyethylene alkyl ether salt, sodium lauryl sulfate, sodium laureth sulfate, Any one or more selected synthetic surfactants may be used. The surfactant may be at least one natural surfactant selected from the group consisting of coco betaine, lecithin, saponin, beeswax, borax, sulphosuccinate, and mixtures thereof. In general, the synthetic emulsifier is more toxic but has excellent emulsifying power. In the present invention, among the synthetic emulsifiers, it is more preferable to use Tween 20, which has high in-vivo safety and emulsion stability.

According to one experimental example of the present invention, the antioxidative capacity of carotenoid nanoparticles including the carotenoid compound represented by the formula (1) or the carotenoid compound represented by the formula (2) and the previously known beta carotene (β-carotene) In the case of conventional beta carotene compounds, antioxidative ability is low regardless of whether it is a nanoparticulate form or a compound itself, whereas the BAS carotinoid compound and the BTS carotenoid compound as the carotinoid compound of the present invention are converted into nanoparticles, Was increased.

In addition, according to one experimental example of the present invention, general carotenoid compounds are antioxidant and lose their function in the early stage, whereas BAS carotenoid nanoparticles and BTS carotenoid nanoparticles of the present invention are in the nanoparticle form, And the efficacy was further increased. In addition, feeding of BAS carotenoid nanoparticles and BTS carotenoid nanoparticles to the nematode C. elegans showed physiological activities such as elongation of body length and reduction of free radical scavenging, and the same antioxidative efficacy was also confirmed in the in vivo experiment.

In terms of stability, the BAS carotenoid nanoparticles and BTS carotenoid nanoparticles of the present invention retained their shape without change in size even after prolonged storage (4 weeks). When exposed to light and heat, BAS carotenoid compounds and BTS carotenoid compounds It was confirmed that the reduction of the number of nanoparticles was mitigated while the decrease in the number of nanoparticles was reduced, thereby preventing the deterioration of function due to light and heat.

Therefore, the carotenoid nanoparticles of the present invention can be safely absorbed in vivo and can be transported to a place where the oxidative stress is present in a complete form in vivo, and can also be used as a carrier for nutrients, drugs, cosmetics, etc. have.

The carotenoid nanoparticles comprising the novel carotenoid compound as particles having a diameter of 1000 nm or less according to the present invention are remarkably improved in terms of stability and functionality as compared with nanoparticles prepared from existing beta carotene compounds. Specifically, in the case of conventional beta-carotene compounds, antioxidative ability is low regardless of the form of nanoparticles or the compound itself, while the carotenoid compound of the present invention has the effect of enhancing the antioxidative function by becoming nanoparticles.

1 shows particle sizes of carotenoid compound (BAS carotinoid) nanoparticles represented by Chemical Formula 1, carotenoid compound (BTS carotenoid) nanoparticles represented by Chemical Formula 2, and beta carotene nanoparticles.
Figure 2 shows UV / Vis spectra before and after production of BAS carotenoid nanoparticles and nanoparticles of BTS carotenoid nanoparticles.
Fig. 3 shows a scanning electron microscopic photograph of BAS, BTS, and beta-carotene nanoparticles.
Fig. 4 shows the increase in antioxidative activity of BAS carotenoids and BTS carotenoid compounds by nanoparticle formation.
FIG. 5 is a graph showing an extended antioxidative activity period of BAS and BTS carotenoid compounds by nanoparticle formation.
Figure 6 shows the increase in stability due to nanoparticle formation of BAS carotenoid nanoparticles, BTS carotenoid nanoparticles and beta carotene nanoparticles.
Figure 7 shows the reduction of photochemical reactions by nanoparticles of BAS carotenoid nanoparticles and BTS carotenoid nanoparticles.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided to further understand the present invention, and the present invention is not limited by the examples.

Manufacturing example  One: BAS  Preparation of carotenoids

Preparation of BAS carotenoids is described in Juwan Maeng, Soo Bong Kim, Nam Joo Lee et al., Conductance Control in Stabilized Carotenoid Wires, Chem. Eur. J. 2010, 16, 7395-7399 ", and specifically prepared according to the following reaction formula.

1-1. (E) -2,7- Bis - (p- 메틸oxyphenyl ) - oct -4- enedioic acid , diethyl ester  (11a)

A 1.6 M hexane solution of n-BuLi (7.48 mmol, 4.70 mL, 3.2 equiv) was added to a THF solution (10 mL) of diisopropylamine (8.18 mmol, 1.16 mL, 3.5 equiv) at 0 ° C. The mixture was stirred at 0 &lt; 0 &gt; C for 20 min and cooled to -78 &lt; 0 &gt; C. A THF solution (5 mL) of ethyl 4-methoxyphenylacetate (7.01 mmol, 1.30 mL, 3.0 equiv) was added to the mixture. The mixture was stirred at -78 <0> C for 40 min and a THF solution (5 mL) of 1,4-dibromo-2-butene (2.34 mmol, 0.50 g, 1.0 equiv) was added. Was stirred at -78 &lt; 0 &gt; C for 1 hour and the mixture was heated to ambient temperature and quenched with 1 M HCl solution. The mixture was extracted with CH ₂ Cl ₂ , washed with 1M HCl, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product SiO ₂ The mixture was purified by flash column chromatography (Hexanes: EtOAc = 17: 3) to obtain diester 11a (1.92 mmol, 0.84 g, a 1: 1 mixture of diastereomers) in 82% yield.

Rf = 0.40 (Hexanes: EtOAc = 4: 1)

^{1 H NMR δ 1.19 (t,} J = 7.1 Hz, 6H), 2.30-2.43 (m, 2H), 2.60-2.73 (m, 2H), 3.44 (dt, Jd = 4.3, Jt = 7.7 Hz, 2H), 2H), 6.80-6.87 (m, 4H), 7.13-7.21 (m, 4H) ppm (m,

¹³ C NMR? 14.1, 36.4 (36.5), 51.0 (51.0), 55.2, 60.6, 113.9, 128.9 (128.9), 129.3 (129.4), 130.8 (130.8), 158.7, 173.6 ; IR (KBr) 2978, 2837, 1730, 1611, 1513, 1250, 1178, 1035 cm ^-1 HRMS (FAB ⁺ ) calcd for C ₂₆ H ₃₃ O ₆ 441.2277, found 441.2280.

*: The ¹³ C NMR peak in parentheses is derived from the diastereoisomer.

1-2. (E) -2,7- Bis - (p- 메틸oxyphenyl ) - oct -4- ene -1,8- diol  (14a)

A THF solution (5 mL) of Diester 11a (3.49 mmol, 1.54 g, 1.0 equiv) was added to a THF suspension of LiAlH ₄ (6.54 mmol, 0.25 g, 2.0 equiv). Stir for 30 min, heat the mixture to ambient temperature and quench with 1 M HCl solution. The mixture is extracted with EtOAc and washed with 1M HCl, dried over anhydrous Na ₂ SO _4, filtered and concentrated under reduced pressure. The crude product was purified by SiO ₂ flash column chromatography (Hexanes: EtOAc = 13: 7) to give diol 14a (3.20 mmol, 1.14 g, a 1: 1 mixture of stereoisomers) in 92% yield.

Rf = 0.14 (Hexanes: EtOAc = 3: 2)

^{1 H NMR δ 1.36 (br s} , 2H), 2.13-2.38 (m, 4H), 2.61-2.80 (m, 2H), 3.53-3.70 (m, 4H), 3.79 (d, J = 0.8 Hz, 6H) , 5.25-5.34 (m, 2H), 6.81-6.89 (m, 4H), 7.00-7.09 (m, 4H) ppm

¹³ C NMR? 35.5 (35.6), 47.6 (47.6), 55.2, 66.8 (66.9), 113.9, 128.9 (128.9), 129.7 (129.7), 134.0, 158.3 ppm; IR (KBr) 3365, 2931, 1611, 1513, 1459, 1248, 1036, 830 cm ^-1 HRMS (FAB ⁺ ) calcd for C ₂₂ H ₂₈ O ₄ 356.1988, found 356.1996.

*: The ¹³ C NMR peak in parentheses is derived from the diastereoisomer.

1-3. (E) -2,7- Bis - (p- 메틸oxy - phenyl ) - oct -4- enedial  (5a)

Oxalyl chloride (3.50 mmol, 0.30 mL, 3.5 equiv) was added to a CH ₂ Cl ₂ solution (10 mL) of DMSO (3.80 mmol, 0.29 g, 3.8 equiv) at -78 ° C. The mixture was stirred for 15 min and a CH ₂ Cl ₂ solution (5 mL) of diol 14a (1.00 mmol, 0.36 g, 1.0 equiv) was added. The mixture was stirred at 8 ℃ for 40 minutes, followed by addition of _{Et 3 N (10.00 mmol, 1.40} mL, 10 equiv). Stir at-78 C for 20 min, heat the mixture to room temperature and quench with 1 M HCl solution. The mixture was extracted with CH ₂ Cl ₂ , washed with 1M HCl, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by SiO ₂ flash column chromatography (Hexanes: EtOAc = 4: 1) to give dial 5a (0.85 mmol, 0.30 g, a 1: 1 mixture of stereoisomers) in 85% yield.

Rf = 0.52 (Hexanes: EtOAc = 3: 2)

^{1 H NMR δ 2.26-2.40 (m,} 2H), 2.62-2.73 (m, 2H), 3.42 (ddd, J = 8.4, 8.0, 7.0 Hz, 2H), 3.81 (d, J = 2.0 Hz, 6H), 2H), 6.85-6.91 (m, 4H), 6.98-6.05 (m, 4H), 9.58

¹³ C NMR 隆 32.7 (32.7), 55.1, 58.0 (58.1), 114.3, 114.7, 127.4, 129.1, 129.8, 158.9, 158.9, 200.2) ppm;

IR (KBr) 2933, 1722, 1609, 1512, 1366, 1250, 1032, 829 cm ^-1

^{HRMS (FAB +) calcd for C} 22 H 24 O 4 352.1675, found 352.1687.

*: The ¹³ C NMR peak in parentheses is derived from the diastereoisomer.

1-4. (LS, 2R) -2- Methyl -1- (4- ( 메틸thio ) phenyl ) -2- (phenylsulfonylmethyl) -3-buten-1-ol (10)

(3.93 g, 24.52 mmol, 1.2 equiv) and Indium (2.58 g, 22.47 mmol, 1.1 equiv) were added to a solution of 4-chloro-2-methyl-2-butene- 5.0 g, 20.43 mmol, 1.0 equiv) in THF (10 mL) and H ₂ O (40 mL). The mixture was heated at 80 &lt; 0 &gt; C for 2.5 hours and cooled to room temperature. The mixture was extracted with EtOAc (30 mL x 3), washed with 1 M HCl (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by silica gel flash column chromatography to obtain homoallylic alcohol 10 (6.72 g, 18.54 mmol) in 91% yield.

^{1 H NMR δ 1.26 (s,} 3H), 2.45 (s, 3H), 3.01 (br s, 1H), 3.14 (A of ABq, JAB = 14.1 Hz, 1H), 3.51 (B of ABq, JAB = 14.1 Hz (D, J = 17.5 Hz, 1H), 4.89 (d, J = 17.5 Hz, 1H) 7.12-7.22 (m, 4H), 7.50-7.66 (m, 3H), 7.85-7.92 (m, 2H) ppm

¹³ C NMR δ 15.6, 19.6, 46.2, 62.5, 78.1, 115.9, 125.5, 127.7, 128.5, 129.2, 133.6, 136.5, 137.9, 139.7, 141.2 ppm

IR (KBr) 3502, 1598, 1447, 1305, 1147 cm &lt; ^-1 &gt;

^{HRMS (FAB +) m / z} cacld for C 19 H 23 O 5 S 363.1266, found 363.1265.

Data for syn: ¹ H NMR δ 1.12 (s, 3H), 2.46 (s, 3H), 6.21 (dd, J = 17.4, 10.8 Hz, 1H) ppm.

1-5. Methyl (4 - ((1E, 3E) -4- methyl -5- ( phenylsulfonyl ) -1,3-pentadienyl) phenyl) sulfane (4)

(2.94 g, 18.33 mmol, 1.1 equiv) and 10-camphorsulfonic acid (4.34 g, 18.33 mmol, 1.1 equiv) were added to a benzene solution of homoallylic alcohol 10 (6.04 g, 16.66 mmol, 1.0 equiv) 50 mL). The mixture was heated at 80 &lt; 0 &gt; C for 3.5 hours and cooled to ambient temperature. The mixture was extracted with EtOAc (30 mL x 3), washed with 1 M HCl (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was recrystallized from EtOH to give all- (E) -allylic sulfone 4 (4.55 g, 13.22 mmol) in the form of a white solid in 79% yield.

^{1 H NMR δ 1.95 (s.} 3H), 2.48 (s, 3H), 3.83 (s, 2H), 5.81 (d, J = 10.9 Hz, 1H), 6.34 (d, J = 15.4 Hz, 1H), 6.84 (d, J = 15.4, 10.9 Hz, 1H), 7.17-7.32 (m, 4H), 7.51-7.67 (m, 3H), 7.85-7.89

¹³ C NMR δ 15.7, 17.7, 66.7, 123.3, 124.9, 126.5, 126.9, 128.5, 129.0, 133.6, 133.6, 133.9, 134.6, 138.4, 138.5 ppm

IR (KBr) 1588, 1489, 1446, 1307, 1149 cm &lt; ^{-1 &}

^{HRMS (FAB +) m / z} cacld for C 19 H 21 O 2 S 2 345.0983, found 345.0981.

1-6. (1E, 3E, 9E, 15E, 17E) -7,12-Bis (p-methoxyphenyl) -1,18- bis 메틸thio ) phenyl ) -5,14- bis ( phenylsulfonyl ) -4,15- dimethyl - octadeca -1,3,9,15,17-pentaene-6,13-diol (7a)

A 1.6 M hexane solution of n-BuLi (9.23 mL, 14.77 mmol, 2.6 equiv) was added to a THF solution (50 mL) of allylic sulfone 4 (4.3 g, 12.48 mmol, 2.2 equiv) at -78 < The orange solution was stirred for 40 min and a THF solution (10 mL) of dialdehyde 5a (2.0 g, 5.68 mmol, 1 equiv) was added over 5 min. The mixture was stirred at -78 &lt; 0 &gt; C for 70 min and 1M HCl (20 mL) was added. The reaction mixture was heated to room temperature, extracted with EtOAc (20 mL x 2), washed with 1 M HCl (15 mL x 2), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by silica gel flash column chromatography to give 7a (5.37 g, 5.20 mmol) in 91% yield.

^{1 H NMR δ 1.67 - 1.85 (} s, 6H; CH 3), 2.10 - 2.67 (m, 8H), 2.47 - 2.48 (s, 6H; SCH 3), 3.71 - 3.87 (m, 2H; PhO 2 SCH), 3.74 - 3.77 (s, 6H; OCH 3), 4.40 - 4.61 (m, 2H; HOCH), 4.94 - 5.08 (m, 2H), 5.67 - 5.92 (m, 2H), 6.24 - 6.39 (m, 2H), (M, 4H), 7.16-7.25 (m, 4H), 7.27-7.66 (m, 4H), 7.40-7.52 , 4H), 7.52-7.60 (m, 2H), 7.64-7.84 (m, 4H) ppm

^{13 C NMR (major isomer) δ} 15.9, 16.3, 31.5, 48.2, 55.4, 74.1, 78.3, 113.8, 123.1, 123.6, 126.8, 127.2, 127.6, 129.0, 129.7, 129.8, 130.6, 132.7, 133.9, 134.3, 135.3, 136.5, 138.7, 158.5 ppm

IR (KBr) 3511, 3033, 2921, 2836, 1610, 1587, 1511, 1492, 1446, 1301, 1248, 1178, 1142, 1090, 1035, 967, 912, 732 cm -1

^{HRMS (FAB +) m / z} cacld for C 48 H 53 O 4 S 2 [C 6 0H 65 O 8 S 4 _{- 2 × (C 6 H 6} O 2 S)] 757.3385, found 757.3389.

1-7. Bis - MOM - ether (9a)

Dimethoxymethane (18 mL, 50 equiv) and P ₂ O ₅ (1.68 g, 11.85 mmol, 3 equiv) were added to a CH ₂ Cl ₂ solution (40 mL) of 7a (4.11 g, 3.95 mmol, 1 equiv) . The brown solution was stirred at room temperature for 14 hours and diluted with CH ₂ Cl ₂ (40 mL). The mixture was washed with saturated NaHCO ₃ solution (10 mL x 2), dried over anhydrous K ₂ CO ₃ , filtered and concentrated under reduced pressure. The crude product was purified by silica gel flash column chromatography (deactivated by 1% Et ₃ N) to give bis-MOM-ether 9a (4.30 g, 3.81 mmol) in 96% yield.

^{1 H NMR δ 1.46-1.92 (s,} 6H; CH 3), 2.23-2.87 (m, 6H), 2.46 (s, 6H; SCH 3), 3.27-3.47 (s, 6H, OCH 3), 3.70-3.80 _{(s, 6H; OCH 3)} , 3.82-3.96 (m, 2H; PhO 2 SCH), 4.23-4.36 (m, 2H; MOMOCH), 4.50-4.68 (m, 4H), 5.00-5.20 (m, 2H) (M, 2H), 5.92-6.10 (m, 2H), 6.15-6.46 (m, 2H), 6.54-6.72 m, 4H), 7.23-7.32 (m, 4H), 7.32-7.60 (m, 6H), 7.60-7.80

^{13 C NMR (major isomer) δ} 15.2, 15.9, 32.0, 48.6, 55.4, 57.2, 76.8, 81.6, 98.9, 113.8, 123.2, 126.7, 127.2, 128.1, 128.4, 128.8, 129.0, 129.9, 130.3, 133.1, 134.0, 134.2, 135.0, 138.8, 141.0, 158.5 ppm

IR (KBr) 2923, 2834, 1610, 1588, 1513, 1492, 1304, 1248, 1178, 1145, 1085, 1030, 967, 912, 832, 808, 730 cm -1

HRMS (FAB ⁺ ) m / z cacld for C ₄₈ H ₅₁ O ₃ S ₂ [C ₆₄ H ₇₃ O ₁₀ S ₄ -2 × (C ₆ H ₅ O ₂ S) - 2 × (CH ₃ OCH ₂ ) ₂ O] 739.3280, found 739.3292

1-8. (P-methoxyphenyl) -1,18-bis (p-methoxyphenyl) -1,2,3,4-tetrahydronaphthalene (1E, 3E, 5E, 7Z, 9E, 11Z, 13E, 15E, 17E) bis (p- 메틸THiophenyl ) -4,15- dimethyl - octadeca Preparation of -1,3,5,7,9,11,13,15,17-nonaene (BAS)

KOMe (4.6 g, 62.68 mmol, 20 equiv) was added to a solution of bis-MOM-ether 9a (3.54 g, 3.13 mmol, 1 equiv.) In cyclohexane (100 mL) and benzene (20 mL). The mixture was heated at &lt; RTI ID = 0.0 &gt; 80-90 C &lt; / RTI &gt; for 17 hours and cooled to room temperature. Most of the solvent was removed under reduced pressure. The mixture was diluted with CH ₂ Cl ₂ , washed with 1 M HCl and H ₂ O, dried over anhydrous K ₂ CO ₃ , filtered and concentrated under reduced pressure. The crude product (2.08 g, 2.77 mmol, 88% yield) was recrystallized from THF and EtOH to give red solid all-trans carotene BAS (1.3 g, 1.8 mmol) in 58% yield.

^{1 H NMR δ 1.98 (s,} 6H), 2.47 (s, 6H), 3.88 (s, 6H), 6.02 (d, J = 15.6 Hz, 2H), 6.10 (d, J = 11.6 Hz, 2H), 6.21 J = 15.2 Hz, 2H), 6.96 (d, J = 8.4 Hz, 2H), 6.30 (m, 2H), 6.30-6.37 J = 8.8 Hz, 4H), 7.17 (d, J = 8.4 Hz, 4H), 7.30 (d, J = 8.4 Hz, 4H) ppm

¹³ C NMR δ 13.0, 16.0, 55.5, 113.9, 125.2, 126.9, 126.9, 130.2, 131.3, 132.1, 132.2, 132.7, 132.8, 133.2, 135.0, 136.4, 136.5, 137.8, 143.0, 159.1 ppm

IR (KBr) 3031, 2920, 1608, 1509, 1491, 1435, 1288, 1246, 1176, 1093, 1033, 965, 909, 836, 805, 732 cm -1

UV (c = 5 x 10 ^-6 M in CH ₂ Cl ₂ )? _Max (?) 464 (126,000), 492 (160,000), 526

HRMS (FAB ⁺ ) m / z calcd for C ₄₈ H ₄₈ O ₂ S ₂ 720.3096, found 720.3085

Manufacturing example  2: BTS  Preparation of carotenoids

BAS carotenoids are described in the paper 'Juwan Maeng, Soo Bong Kim, Nam Joo Lee et al., Conductance Control in Stabilized Carotenoid Wires, Chem. Eur. J. 2010, 16, 7395-7399 &quot;, and specifically prepared according to the following reaction formula.

2-1. (E) -2,7- Di -p- tolyl - oct -4- enedioic acid , diethyl ester (11b)

A solution of LDA (32.73 mmol, 4.67 mL) and 1.6 M hexane solution of n-BuLi (29.92 mmol, 0.35 mmol) was added dropwise to a THF solution (10 mL) of ethyl p-tolylacetate (23.38 mmol, 4.21 mL) 18.70 mL) in THF (25 mL )] and then reacted, at -78 ℃ for 1 hour, 1,4-dibromo-2-butene (9.35 mmol, 2.00 g, react with 1.0 eq), SiO ₂ column The residue was purified by chromatography (Hexanes: EtOAc = 19: 1) to give diester 11b (7.47 mmol, 3.05 g, a 1: 1 mixture of stereoisomers) in 80% yield.

Rf = 0.80 (Hexanes: EtOAc = 4: 1)

^{1 H NMR δ 1.17 (dt,} Jd = 1.6, Jt = 7.2 Hz, 6H), 2.30 (d, J = 2.4 Hz, 6H), 2.32-2.42 (m, 2H), 2.63-2.76 (m, 2H), 2H), 7.06-7.16 (m, 8H) ppm (m, 2H), 3.46 (ddd, J = 6.0,5.7,5.4 Hz, 2H), 3.99-4.15

¹³ C NMR δ 14.3, 21.1, 36.9, 51.7, 60.6, 128.0, 129.3 (129.4) *, 129.7, 136.2 (136.2) *, 136.5 (136.7) *, 173.3

IR (KBr) 2981, 1733, 1514, 1154, 1037, 973, 820 cm -1

HRMS (CI ⁺ ) calcd for C ₂₆ H ₃₃ O ₄ 409.2379, found 409.2379

*: The ¹³ C NMR peak in parentheses is derived from the diastereoisomer.

2-2. (E) -2,7- Di -p- tolyl - oct -4- ene -1,8- diol (14b)

A THF solution (20 mL) of LiAlH ₄ (15.88 mmol, 0.63 g) was reacted with diester 11b (7.56 mmol, 3.09 g) at 0 ° C for 30 minutes and then subjected to SiO ₂ column chromatography (Hexanes: EtOAc = 13: To give diol 14b (6.78 mmol, 2.20 g, a 1: 1 mixture of stereoisomers) in 90% yield.

Rf = 0.42 (Hexanes: EtOAc = 3: 2)

^{1 H NMR δ 1.43 (br s} , 2H), 2.15-2.36 (m, 4H), 2.32 (s, 6H), 2.70 (tt, J = 5.9, 5.2 Hz, 2H), 3.54-3.68 (m, 4H) , 5.24-5.38 (m, 2H), 6.98-7.04 (m, 4H), 7.08-7.13 (m, 4H) ppm

¹³ C NMR? 21.3 (21.4) *, 35.8 (35.8) *, 48.3 (48.4) *, 66.9 (67.0) *, 128.2, 129.5, 130.0

IR (KBr) 3280, 2919, 1513, 1441, 1376, 1066, 1050, 1021, 978, 815, 735 cm ^-1

^{HRMS (CI +) calcd for C} 22 H 29 O 2 325.2167, found 325.2167

*: The ¹³ C NMR peak in parentheses is derived from the diastereoisomer.

2-3. (E) -2,7- Di -p- tolyl - oct -4- enedial (5b)

(13.82 mmol, 1.08 g), oxalyl chloride (6.91 mmol, 0.62 mL), and Et ₃ N (31.40 mmol, 4.36 mL) in CH ₂ Cl ₂ (3.14 mmol, 1.02 g) 15 mL) and purified by SiO ₂ column chromatography (Hexanes: EtOAc = 9: 1) to give dialb 5b (2.65 mmol, 0.85 g, a 1: 1 mixture of stereoisomers) in 84% yield.

Rf = 0.81 (Hexanes: EtOAc = 3: 2)

^{1 H NMR δ 2.28-2.40 (m,} 2H), 2.35 (s, 6H), 2.64-2.73 (m, 2H), 3.42 (ddd, J = 7.8, 6.0, 5.7 Hz, 2H), 5.30-5.38 (m , 2H), 6.94-7.02 (m, 4H), 7.10-7.20 (m, 4H), 9.59

¹³ C NMR? 21.3 (21.3) *, 33.0 (33.1) *, 58.8 (58.9) *, 129.0 (129.0) *, 129.4 (129.5) *, 129.9, 132.9

IR (KBr) 2921, 1721, 1606, 1514, 1440, 1039, 813 cm ^-1

^{HRMS (CI +) calcd for C} 22 H 25 O 2 321.1854, found 321.1852

*: The ¹³ C NMR peak in parentheses is derived from the diastereoisomer.

2-4. (1E, 3E, 9E, 15E, 17E) -1,18-Bis (p- (methylthio) bis ( phenylsulfonyl ) -4,15- dimethyl -7,12- di -p- tolyl - octadeca -1,3,9,15,17-pentaene-6,13-diol (7b)

The allylic sulfone 4 (2.15 g, 6.23 mmol, 2.5 equiv), which had been treated with n-BuLi (1.6 M solution in hexane, 4.5 mL, 7.13 mmol, 2.9 equiv) in THF (50 mL) (2.31 g, 2.28 mmol) was obtained in 92% yield by reaction with dialdehyde 5b (0.80 g, 2.49 mmol, 1 equiv) in tetrahydrofuran (10 mL) at -78 ° C for 1 hour.

^{1 H NMR δ 1.65-1.85 (s,} 6H; CH 3), 1.90-2.70 (m, 6H), 2.24-2.33 (s, 6H; CH 3), 2.43-2.52 (s, 6H; SCH 3), 3.18 _{-4.04 (m, 2H; PhO 2} SCH), 3.30-3.70 (br s, 2H; OH), 4.40-4.64 (m, 2H; HOCH), 4.90-5.12 (m, 2H), 5.66-5.95 (m, 2H), 6.20-6.38 (m, 2H), 6.44-6.87 (m, 2H), 6.87-7.09 (m, 8H), 7.15-7.25 (m, 4H), 7.25-7.38 7.50 (m, 4H), 7.50-7.63 (m, 2H), 7.63-7.88 (m, 4H) ppm

IR (KBr) 3513, 3021, 2921, 1589, 1513, 1492, 1446, 1303, 1293, 1142, 1084, 967, 911, 730, 688 cm -1

HRMS (FAB ⁺ ) m / z cacld for C ₄₈ H ₅₃ O ₂ S ₂ [C ₆₀ H ₆₅ O ₆ S ₄ -2 × (C ₆ H ₆ O ₂ S)] 725.3487, found 725.3475

2-5. Bis - MOM - ether (9b)

7b (2.20 g, 2.22 mmol, 1 equiv) and dimethoxymethane (9.83 mL, 50 equiv) of _{P 2 O 5 (0.90 g,} 6.65 mmol, 3 equiv) under CH ₂ Cl ₂ 18 at room temperature in (50 mL) For 2 hours to give bis-MOM-ether 9b (2.24 g, 2.04 mmol) in 92% yield.

^{1 H NMR δ 1.46-1.95 (s,} 6H; CH 3), 2.02-2.96 (m, 6H), 2.20-2.36 (s, 6H, CH 3), 2.35-2.53 (s, 6H; SCH 3), 3.20 _{-3.52 (s, 6H; OCH 3} ), 3.72-3.98 (m, 2H; PhO 2 SCH), 4.22-4.39 (m, 2H; MOMOCH), 4.40-4.72 (m, 4H), 4.83-5.36 (m, 2H), 5.66-6.12 (m, 2H), 6.16-6.41 (m, 2H), 6.45-6.85 (m, 2H), 6.85-7.10 (m, 8H), 7.11-7.22 7.33 (m, 4H), 7.33-7.61 (m, 6H), 7.62-7.88 (m, 4H) ppm

IR (KBr) 3021, 2921, 1589, 1514, 1492, 1304, 1144, 1085, 1026, 967, 911, 808, 732, 689 cm -1

^{HRMS (FAB +) m / z} cacld for C 48 H 51 OS 2 [C 64 H 73 O 8 S 4 - 2 × (C 6 H 5 O 2 S) - 2 × (CH 3 OCH 2) - H 2 O ] 707.3381, found 707.3400

2-6. (1E, 3E, 5E, 7Z, 9E, 11Z, 13E, 15E, 17E) Bis (p- 메틸THiophenyl ) -4,15-dimethyl-7,12-di-p-tolyloctadeca-1,3,5,7,9,11,13,15,17-nonaene (BTS)

bis-MOM-ether 9b (1.60 g, 1.45 mmol) and KOMe (2.04 g, 29.15 mmol, 20 equiv) were reacted in cyclohexane (50 mL) and benzene (10 mL) A crude carotene product (0.75 g, 1.08 mmol, 74% yield) was obtained which was recrystallized from THF and EtOH to give red solid all-trans carotene BTS (0.52 g, 0.75 mmol) in 52% yield.

^{1 H NMR δ 1.97 (s,} 6H), 2.43 (s, 6H), 2.47 (s, 6H), 6.01 (d, J = 15.2 Hz, 2H), 6.10 (d, J = 11.2 Hz, 2H), 6.18 J = 15.2 Hz, 2H), 6.50 (d, J = 15.6 Hz, 2H), 7.06 (dd, J = 8.0 Hz, 2H), 7.07 (d, J = 6.8 Hz, 4H), 7.17 (d, J = 7.6 Hz, 4H), 7.23 4H) ppm

IR (KBr) 3031, 2920, 1608, 1509, 1491, 1435, 1093, 965, 816, 617 cm ^-1

UV (c = 5 x 10 ^-6 M in CH ₂ Cl ₂ )? _Max (?) 464 (96,000), 491 (128,000), 526 (105,000) nm

HRMS (FAB ⁺ ) m / z calcd for C ₄₈ H ₄₉ S ₂ 689.3276, found 689.3285

Example  One: BAS  Production of carotenoid nanoparticles

BAS carotenoids were added to acetone, which had been pre-warmed at 50 ° C, using a thermostatic water bath, and the mixture was dissolved for 1 minute through a magnetic stirrer.

The carotenoid solution dissolved in the acetone solution was transferred to a brown buret and dropped at a rate of about 0.4 mL / sec into a 1% Tween 20 aqueous solution which had been warmed to 50 캜. At this time, the ratio of the aqueous solution to the organic solution was 1: 9, and when mixed, the solution was magnetically stirred to ensure smooth dispersion of the carotenoids. The time until mixing with the aqueous solution was limited to 5 minutes.

The mixture of carotenoid-dissolving acetone and aqueous solution was treated at 70 캜 for 30 minutes in a vacuum distiller to remove acetone, and the remaining impurities were removed by 0.2 탆 filtering.

Example  2: BTS  Production of carotenoid nanoparticles

The BTS carotenoids corresponding to 1 mM were added to the acetone, which had been heated to 50 ° C, using a thermostatic water bath, and the mixture was dissolved for 1 minute through a magnetic stirrer immediately after the addition.

The carotenoid solution dissolved in the acetone solution was transferred to a brown buret and dropped at a rate of about 0.4 mL / sec into a 1% Tween 20 aqueous solution which had been warmed to 50 캜. At this time, the ratio of the aqueous solution to the organic solution was 1: 9, and when mixed, the solution was magnetically stirred to ensure smooth dispersion of the carotenoids. The time until mixing with the aqueous solution was limited to 5 minutes.

The mixture of carotenoid-dissolving acetone and aqueous solution was treated at 70 캜 for 30 minutes in a vacuum distiller to remove acetone, and the remaining impurities were removed by 0.2 탆 filtering.

Example  3: Preparation of beta-carotene nanoparticles

Beta-carotene (purchased from Wako Pure Chemical, Tokyo, Japan), which corresponds to 1 mM, was added to acetone, which had been warmed to 50 ° C, using a thermostatic water bath, and the mixture was dissolved for 1 minute through a magnetic stirrer.

The carotenoid solution dissolved in the acetone solution was transferred to a brown buret and dropped at a rate of about 0.4 mL / sec into a 1% Tween 20 aqueous solution which had been warmed to 50 캜. At this time, the ratio of the aqueous solution to the organic solution was 1: 9, and when mixed, the solution was magnetically stirred to ensure smooth dispersion of the carotenoids. The time until mixing with the aqueous solution was limited to 5 minutes.

The mixture of carotenoid-dissolving acetone and aqueous solution was treated at 70 캜 for 30 minutes in a vacuum distiller to remove acetone, and the remaining impurities were removed by 0.2 탆 filtering. Beta carotene nanoparticles were obtained in 83% yield by the above method.

Experimental Example  1: Analysis of nanoparticle size

In order to examine the sizes of the carotenoid nanoparticles prepared in Examples 1 to 3, analysis was carried out using a nanoparticle analyzer (HPPS, Malvern Instrument, England) loaded with Dynamic Light Scattering (DLS) Is shown in Fig.

Fig. 1 (a) shows that the size of the BAS nanoparticles is about 80 nm in diameter. Fig. 1 (b) shows the size of BTS nanoparticles, and it was confirmed that they had a diameter of about 90 nm. 1 (c) shows the size of beta-carotene nanoparticles (purchased from Wako Pure Chemical, Tokyo, Japan) and has a diameter of about 10 nm. 1 (a), (b) and (c), it was confirmed that nanoparticles having a diameter of several tens of nanometers (nm) were produced, and that nanoparticles of different sizes were formed for each of the carotenoids there was.

Experimental Example  2: UV / Vis  Spectrum analysis

The UV / Vis spectral change of the carotenoid nanoparticles prepared in Examples 1 and 2 was analyzed using a spectrophotometer (UV / Vis spectrophotometer, Thermo Scientific Inc, Germany), and the absorbance at 200 nm to 800 nm was analyzed. The results are shown in Fig.

It was confirmed that the maximum absorption wavelength was changed by each BAS carotenoid and BTS carotenoid having a nanoparticle form.

Experimental Example  3: Scanning Electron Microscopy Analysis

The surface morphology and microstructure of the carotenoid nanoparticles prepared in Examples 1 to 3 were analyzed by Scanning Electron Microscopy (SEM). The results obtained are shown in FIG.

 FIG. 3 shows that the nanoparticles were spherical in shape of aggregation of carotenoid molecules in the aqueous phase as a result of several superposition processes for morphological analysis.

Experimental Example  4: Antioxidant ability  analysis

DPPH (1,1-diphenyl-2-picryl hydrazyl) radical scavenging activity was examined to see whether the BAS carotenoids and BTS carotenoid nanoparticles prepared in Examples 1 and 2 improved the antioxidative function. 4 and Fig. 5, respectively.

FIG. 4 shows a comparison of the antioxidative capacity of carotenoids in a normal aqueous solution with the antioxidative capacity in the case of nanoparticles. Herein, the term "water" refers to carotenoids in an aqueous solution which has not been converted into nanoparticles. To make the nanoparticles and nanoparticles have the same size and concentration conditions, 9 ml of distilled water was mixed with 1 ml of an organic solvent containing 1 mM carotenoid and subjected to ultrasonication treatment.

As a result, it was confirmed that the novel carotenoids BAS and BTS exhibit an antioxidative activity in the non-nanoparticle state while the radical scavenging ability is enhanced in the nanoparticle state (NP).

FIG. 5 shows the results of confirming radical scavenging ability of the nanoparticles over time. As a result, it was confirmed that the antioxidant activity was improved over a long period of time without decreasing the activity over time.

Experimental Example  5: Storage stability analysis

To evaluate the storage stability of BAS carotenoids and BTS carotenoid nanoparticles prepared in Examples 1 and 2, the size of the nanoparticles stored on the day of manufacture and the nanoparticles stored for 4 weeks were measured using a nanoparticle analyzer (HPPS, Malvern instrument, England) , And the results obtained are shown in Fig.

6 (a), 6 (b) and 6 (c) show that there is no significant difference in size between immediately after production of BAS carotenoid nanoparticles, BTS carotinoid nanoparticles, and beta carotene nanoparticles and after storage for 4 weeks, It was confirmed that the novel carotenoid-type nanoparticles according to the present invention were stably dispersed in the aqueous solution without aggregation of the carotenoids despite the storage for a long period of time.

Experimental Example  6: Exposure stability analysis

To analyze the reduction of the number of effective carotenoids when BAS carotenoids and BTS carotenoid nanoparticles prepared in Examples 1 and 2 were exposed to light and heat, non-nanoparticles and nanoparticles were exposed to direct sunlight and heat for one hour The DPPH (1,1-diphenyl-2-picryl hydrazyl) radical scavenging ability was confirmed, and the obtained results are shown in Fig.

FIG. 7 (a) shows the radical scavenging ability of non-nanoparticles after exposure to stimulation, and it was confirmed that all of the effective carotenoids having antioxidant activity were degraded. Fig. 7 (b) shows the results of the radical scavenging ability after nanoparticle stimulation. It was confirmed that the number of effective carotenoids was reduced after stimulation but was considerably larger than that of non-nanoparticles.

Claims (5)

Comprising nanoparticles of a carotenoid compound represented by the following formula (2) into particles having a diameter of 1000 nm or less,
Methods for improving antioxidant potential, or light or thermal stability, of carotenoids versus non-nanoparticulated carotenoids:
(2)

.
A carotenoid nanoparticle comprising a carotenoid compound represented by the following formula (2) as particles having a diameter of 1000 nm or less,
Compositions for improving the antioxidant ability, or the optical or thermal stability, of carotenoids versus non-nanoparticulate carotenoids:
(2)

.
3. The composition of claim 2, wherein the nanoparticles have a diameter of from 1 nm to 100 nm.
The nanoparticle as set forth in claim 2, wherein the nanoparticle is selected from the group consisting of sodium linear alkylbenzenesulfonate, sodium alpha-sulfonate, sodium alpha-olefinsulfonate, sodium alkyl ether esterate, polyoxyethylene alkyl ether salt, sodium lauryl sulfate, sodium laureth sulfate, &Lt; / RTI &gt; wherein the composition further comprises one or more synthetic surfactants selected from the group consisting of a mixture of a surfactant and a surfactant.
The method according to claim 2, wherein the nanoparticles further comprise at least one natural surfactant selected from the group consisting of coco betaine, lecithin, saponin, beeswax, borax, sulphosuccinate, and mixtures thereof &Lt; / RTI &gt;

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