實驗室先前研究口腔鱗狀細胞癌SCC-15與基質金屬蛋白酶(Matrix metalloproteinase-9, MMP-9)，以SPR證明了強力黴素(doxycycline, DOX)是與TGF-β/Smad pathway中的Smad4蛋白結合，以抑制MMP-9的基因表現，並藉著移除DOX四號碳上的二甲基胺生成TMC-1，降低其抗菌性，但保留甚至提升其抑制膠原蛋白酶活性的能力。本研究希望利用化學修飾TMC-1能進一步提升藥物抑制效果，有文獻結果顯示親油性及推電子基團可能會提升四環黴素的抑制效果，於是本研究在9號碳加上第三丁基、於7號碳加上硝基並還原成胺基，合成TMC-4、TMC-5及TMC-6，並利用HPLC和NMR純化和鑑定產物，再以Zymography測試各藥物對SCC-15的MMP-9抑制效果。結果顯示在C7引入強拉電子的硝基時藥物抑制效果下降，但毒殺能力也上升，而改引入推電子的胺基時，其抑制能力在2.5 μg/ml時觀察不到，但藥物濃度上升後重新展現明顯的抑制效果，而在C9引入第三丁基後，雖然MMP-9表達大幅下降，但藥物毒殺能力也上升了，另外也發現強力黴素衍生物能抑制MMP-2表達，其趨勢與MMP-9相近。另外我們也發現四環黴素類化合物對正常細胞的毒性較對癌細胞低。TMC-1依然是這些藥物中抑制MMP-9表達最明顯的，在2.5 μg/ml就有明顯效果。

　　The laboratory previously studied oral squamous cell carcinoma SCC-15 and matrix metalloproteinase-9 (MMP-9), and demonstrated by SPR that doxycycline (DOX) combines with Samd4 in TGF-β/Smad pathway to inhibit the gene expression of MMP-9. Removing the dimethylamine on C4 of DOX, which called TMC-1, will reduce its antibacterial, but retain or even enhance its effect of inhibiting the activity of collagenase. This study try to further promote the doxycycline analogs inhibition effect by chemical modification of 4-dedimethylaminodoxycycline (TMC-1). Literatures report that lipophilicity and electron-donating groups may enhance the inhibitory effect of tetracycline. Add a t-butyl group on the 9th carbon, a nitro group on the 7th carbon, and then reduce the nitro group to an amine group to produce
9-t-butyl-4-dedimethylaminodoxycycline(TMC-4),
9-t-butyl-4-dedimethylamino-7-nitrodoxycycline (TMC-5)
and 7-amine-9-t-butyl-4-dedimethylaminodoxycycline (TMC-6).
　　The products were purified and identified by HPLC and NMR. Then test the compounds inhibitory effect by zymography. Nitro group, which is a strong electron-withdrawing group, on C7 lowered its inhibition while enhanced its cytotoxicity. Samples of TMC-4, which was hydrophobic, show very low activity of MMP-9 but more toxic. The analog with an electron-donating group, amine group, show no ability of inhibition at low concentration but restored an explicit ability of inhibition at higher concentration. TMC-1 is still the most significant inhibitor of MMP-9 expression in these drugs, and has a considerable effect at concentration of 2.5 μg/ml.

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