| Lacquer is a traditional natural coating material.Urushiol is the main compound of lacquer,and large amounts of studies mainly focus on urushiol self-polymerization,metal polymerization,the chemical and mechanical properties of urushiol organic polymers and other functional materials.It has the active group structure of phenolic hydroxyl group and side chain unsaturated monoene or diene.Besides,literatures report that urushiol has a good biological activity in inhibiting tumor,but it is allergic.How to synthesize urushiol small molecule derivatives with low toxicity and high efficiency against tumor by urushiol active group is a difficult point in the world.In this paper,C15 triene urushiol was isolated and purified from raw lacquer,and C15 triene urushiol Pechmann,D-A,halogen,mercaptan long chain alkane and phenylboric acid were obtained by chemical organic synthesis.Urushiol derivatives such as amino sulfoxide and triazole and urushiol based macromolecular drugloaded micellar materials were evaluated for their antitumor activity and safety.It will provide a theoretical and practical basis for the development of urushiol derivatives and urushiol based micellar materials.The main contents of the paper are as following:1.C15 monoene urushiol(2a)and C15 triene urushiol(2b)were isolated and purified from raw lacquer.C15 triene urushiol Pechmann derivative(2c)was synthesized from urushiol aryl 1 active phenolic hydroxyl group and ethyl acetoacetate under acidic conditions.The chemical structure of C15 urushiol and its Pechmann derivatives were characterized.C15 urushiol and its urushiol Pechmann derivative with rigid benzene ring skeleton,side chain C15 olefine and coumarin structure effectively suppressed the growth of Hep G2 cells(2c > 2a > 2b).The half inhibitory concentration(IC50)was 29.3 μM,55.5 μM and 27.1 μM,respectively.Compound 2a,2b,2c combined with paclitaxel showed obvious synergistic inhibitory effect on the proliferation of Hep G2 cells in vitro.When the ratio of urushiol Pechmann compound 2c to paclitaxel was 6.775 μM and 7.9 ng/m L,the synergistic effect was the strongest.The synergy index(CI)reached 0.641.2.Based on the Diels-Alder [4+2] reaction,eight novel urushiol based D-A derivatives 3e～3l were synthesized by introducing the pentagonal ring skeleton of maleic anhydride with different electronegativity-NH,-O and-F,-Cl,-Br halogens.Their compounds were isolated and the chemical structures were identified.The hexagonal cyclene structure formed in the side chain of urushiol increased the electron cloud density in this region.The inhibitory effects of 12 compounds 3a～3l on the proliferation of 10 common human tumor cells were detected by MTT assay.The general regularity of inhibiting the proliferation of tumor cells was as following: urushiol alkane side chain maleic anhydride compounds containing NHCl(3k)> O-Cl(3h)> NH-Br(3l)> O-F(3g)> O-Br(3i)> O(3e)> NH(3j).The modified urushiol D-A derivative had a good inhibitory effect on human gastric cancer cell line MKN-45 and human colon cancer cell line HCT-116.Among all tested compounds,compounds 3e and 3k performed best in restraining MKN-45 and HCT-116.The values were 3.2 μM and 2.6 μM for MKN-45,and 1.2 μM,and 0.66 μM for HCT-116.In addition,these two could effectively inhibit to ther eight kinds of tumor cells.3.The phenolic hydroxyl groups of C15 triene urushiol were protected by alkylation,Pechmann reaction and boric acid esterification.The atomic diversity and electronegativity of urushiol derivatives were enhanced.Sixteen new C15 triene urushiol derivatives were synthesized and their chemical structures were identified.Long chain alkyl chloride was introduced into the phenolic hydroxyl group of urushiol for compound 4h.The dose of antiHep G2 activity decreased slightly to 37 μM,but the toxicity to LO2 of normal hepatocytes LO2 decreased significantly to 180 μM.Compound 4j was urushiol Pechmann derivative,and its inhibitory activity against Hep G2 basically maintained at 28 μM,but its toxicity to LO2 in normal hepatocytes reduced by 3 times to 4 times,about 180 μM.Compounds 4a,4d and 4i were the products of sulfhydryl,phenylboric acid and amino sulfoxide,respectively,and their IC50 values for inhibiting the proliferation of Hep G2 increased to 7 μM～15 μM,while the IC50 dose concentration for LO2 in normal hepatocytes reduced to 120 μM～150 μM.Among them,the IC50 s of urushiol containing mercaptol derivatives for Hep G2 and LO2 were 7.886 μM and 150 μM,respectively.4.A series of C15 triene urushiol bisphenol hydroxyl protected derivatives were synthesized by alkylation and click chemical synthesis,and their chemical structures were identified.Urushiol triazole compound 5d with rich electronic structure had high electron cloud density and affinity with protein.It also had a high antitumor activity against Hep G2,and IC50 was 2.833 μM.Urushiol bisphenol hydroxyl alkynyl ether compound 5c,5f and urushiol Pechmann phenol hydroxyalkynyl ether compound 5v decreased anti-Hep G2 activity.The cytotoxicity of compound 5j on LO2(IC50=198 μM)was lower than that of urushiol triazole compound 5d(IC50=198 μM).Furthermore,Compound 5j’s IC50 value against LO2 was more than 20 times higher than that against Hep G2,and it was less toxic to LO2 of normal hepatocytes.Therefore,the Pechmann derivatives of urushiol 5v and urushiol triazole compounds 5d could be used as the basic skeleton for the synthesis of urushiol derivatives in the future.5.A kind of p H targeting urushiol micellar BPAU-NH2-Gal-PTX,was prepared by Michael addition,hydrophobic urushiol phenylboric acid derivatives and traditional micellar materials,and its chemistry structure was indentified.The particle size of paclitaxel micelle embedded with BPAU-NH2 and BPAU-NH2-Gal were about 135 nm and 195 nm.The results of paclitaxel release in vitro showed that the more drug entrapment,the stronger the hydrophobicity between drugs and the slower the release rate.When micelle and paclitaxel were dissolved in good solvent DMSO,the entrapment efficiency of micelle against paclitaxel was about 99.5%,and the loading rate of BPAU-NH2-Gal was about 5.0%.In acidic p H environment,the release rates of paclitaxel and urushiol were about 64.1% and 49.5%,respectively.In other word,acid condition(p H 4.5)accelerated the drug release rate,and the drug release behavior of urushiol micellar showed a strong p H dependent type.Toxicological experiments in vitro and in vivo showed that paclitaxel loaded micelle BPAUNH2-Gal-PTX entered Hep G2 cells by endocytosis and released paclitaxel and urushiol through micelle to inhibit the growth of Hep G2 cells.The results indicated that targeted BPAU-NH2-Gal micelle had high uptake and enhanced anti-tumor effect. |
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