| 摘要: | 本研究主要利用有機合成的方法，將不同數量及不同位置的羥基取代苯甲酸化合物與色洛冬寧進行醯胺化反應後，得到一系列的多羥基取代之N-色洛冬寧苯甲醯胺衍生物（化合物1 ~ 10），進行四種不同活性測試方法：一、清除DPPH自由基能力抗氧化活性測試；二、總抗氧化能力活性測試(TEAC)；三、抑制酪胺酸酶活性美白測試；四、紫外線／可見光光譜儀吸收測試，並以水溶性維生素E、熊果素、水楊酸辛酯當作對照組，探討其抗氧化能力、抑制酪胺酸酶的活性能力與吸收紫外線能力，再進一步探討其應用於化粧品上的價值。
在清除DPPH自由基的抗氧化能力結果顯示，相較於對照組水溶性維生素E之IC50值13.04 μg/ml，多羥基取代之N-色洛冬寧苯甲醯胺衍生物（化合物 1 ~ 10），皆具有相當不錯的抗氧化能力，十種醯胺化合物的 IC50值介於8.75 ~ 18.02 μg/ml之間，化合物1、4、5、7、10的抗氧化能力效果皆優於對照組水溶性維生素E的IC50值，其中又以化合物10的效果最佳(IC50值8.75 μg/ml)，而清除DPPH自由基的能力發現與羥基的數量及其位置有關。當苯甲醯基的苯環上羥基取代數量越多，其清除DPPH自由基的能力越好，三羥基效果優於雙羥基、單羥基，雙羥基效果優於單羥基；而苯甲醯基苯環上若為多羥基且羥基的位置為鄰位或對位的關係時，其在清除DPPH自由基的能力較佳。
而在總抗氧化能力結果顯示，相較於對照組水溶性維生素E之IC50值5.23 μg/ml，多羥基取代之N-色洛冬寧苯甲醯胺衍生物（化合物1 ~ 10）之總抗氧化能力效果IC50值介於2.49 ~ 3.92 μg/ml之間，十種醯胺化合物的總抗氧化效果皆比對照組水溶性維生素E的抗氧化效果還要來得優越，其中效果最強的仍為化合物10，其 IC50值為 2.49 μg/ml。
在抑制酪胺酸酶活性結果顯示，相較於對照組熊果素的IC50值1.19 mg/ml，效果優於熊果素的為化合物5及化合物10，而其中又以化合物 5 (IC50值0.18 mg/ml)的美白效果為最佳。
在紫外線可見光吸收測試結果顯示，相較於對照組水楊酸辛酯在UVB波長(280 ~ 320 nm)範圍最大吸收波長為306 nm，其莫耳吸收係數為4.03 × 103 ㎝-1M-1，而多羥基取代之N-色洛冬寧苯甲醯胺衍生物（化合物1 ~ 10）在306 nm的莫耳吸收係數介於4.14 × 103 ~ 11.45 × 103㎝-1M-1，其吸收紫外線的效果皆優於對照組水楊酸辛酯，且其在UVB波長的範圍吸收波長的光譜也較寬廣，可知每個化合物並非只是對UVB波長範圍內的單一波長有吸收度，而是對UVB波長範圍具有較寬廣的光譜吸收。而本研究所合成之多羥基取代N-色洛冬寧苯甲醯胺衍生物（化合物1 ~ 10）莫耳吸收係數最高者為化合物4，其在波長306 nm的莫耳吸收係數達11.43 × 103㎝-1M-1，為對照組水楊酸辛酯的莫耳吸收係數的2.84倍，且化合物4的吸收波長涵遝蒤紖VB波長(280 ~ 320 nm)的範圍，故化合物4能提供UVB波長的最佳防護；另外化合物5除了具有UVB波長的吸收外，還具有UVA波長的吸收作用。
This research utilized the method of organic synthesis, researching different amounts and positions of hydroxy-substituted benzoic acid and serotonin into amide reaction, so as to produce a series of polyhydroxy substituted N-benzoylserotonin derivatives (compound 1-10). Accordingly, this study conducts the following four tests. I. The activity of antioxidant and the experiment of ability to scavenge DPPH free radical, II. The ability of overall antioxidant—the activity of TEAC, III. The activity test of inhibit Tyrosinase, and IV. The study of absorbing UV / VIS Spectrophotometers. This study use trolox, arbutin, and octyl salicylate as the control groups, and to test the ability of anti-oxidation, the activity of inhibit Tyrosinase, and the ability of absorbing UV. This research ultimately intends to investigate the applied value of those compounds into cosmetics.
The results of the ability of anti-oxidation scavenging DPPH free radical shows that polyhydroxy substituted N-benzoylserotonin derivatives (compound 1-10) have the generally better ability of anti-oxidation than control group (trolox), in which the IC50 of compound 1-10 ranges 8.75 to 18.02 μg/ml and the IC50 of trolox is 13.04 μg/ml. The abilities of anti-oxidation of compound 1, 4, 5, 7, and 10 are all better than the control group, trolox. Among all, compound 10 has the best ability of anti-oxidation (IC50 is 8.75 μg/ml). This study also discovers that the ability of scavenging DPPH free radical is strongly related to the number and the position of hydroxyl. The more the number of hydroxyl substitution in phenyl ring, the better the ability of scavenging DPPH free radical. That is, trihydroxy is better than dihydroxy, and dihydroxy is better than monohydroxy. Moreover, when the polyhydroxy exists in phenyl ring and it is ortho-position or para-position, the ability of scavenging DPPH free radical is better too.
In the study of testing the overall ability of anti-oxidation, polyhydroxy substituted N-benzoylserotonin derivatives (compound 1-10) still have the better overall ability of anti-oxidation than control group, trolox. The IC50 values of compound 1-10 range 2.49 to 3.92, while the IC50 value of trolox is 5.23 μg/ml. Compound 10 is still the best derivative which has the best overall ability of anti-oxidation; IC50 value is 2.49 μg/ml.
In the activity study of inhibit Tyrosinase, compound 5 and compound 10 show better ability than control group, arbutin, in which the IC50 value of arbutin is 1.19 mg/ml. Between the two compounds, compound 5 shows the best whitening effect (IC50 value is 0.18 mg/ml).
In the study of UV absorbing test, polyhydroxy substituted N-benzoylserotonin derivatives (compound 1-10) shows better ability to absorb UV and shows wider range of absorbing UVB than control group, octyl salicylate. With respect to octyl salicylate, octyl salicylate can abosorb the highest wavelength up to 306 nm under UVB (wavelength ranges 280 nm ~320 nm), and the ε is 4.03 × 103 ㎝-1M-1. However, the ε of polyhydroxy substituted N-benzoylserotonin derivatives (compound 1-10) under 306 nm range 4.14 × 103 ~ 11.45 × 103㎝-1M-1. It is reasonable that the compounds in this study can not only absorb the individual wavelength amoung the wavelenght of UVB, but also absorb wider range of wavelength. Amoung 10 compounds (polyhydroxy substituted N-benzoylserotonin derivatives), compound 4 shows the highest number of ε. The ε of compound 4 is high up to 11.43 × 103㎝-1M-1, which is the 2.84 times of the ε of control group, octyl salicylate. Moreover, compound 4 can absorb all the wavelength if UVB (280 nm - 320 nm), so compound 4 can provide the best protection from UVB. Additionally, this study also discovers that the compound 5 not only can absorb UVB, but also can absorb UVA.
Keywords: N-Benzoylserotonin, amide reaction, synthesis, Antioxidant, free radical, anti-aging, whitening, Tyrosine, Tyrosinase, UV |
