CN110746384A - Compound extracted from Saururi herba and its application in preventing and treating nasopharyngeal carcinoma - Google Patents
Compound extracted from Saururi herba and its application in preventing and treating nasopharyngeal carcinoma Download PDFInfo
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- CN110746384A CN110746384A CN201910757024.4A CN201910757024A CN110746384A CN 110746384 A CN110746384 A CN 110746384A CN 201910757024 A CN201910757024 A CN 201910757024A CN 110746384 A CN110746384 A CN 110746384A
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- GMTJIWUFFXGFHH-WPAOEJHSSA-N sauchinone Chemical compound C1=C2O[C@@]3(C(OCO3)=CC3=O)[C@H]4[C@H]3C[C@@H](C)[C@H](C)[C@H]4C2=CC2=C1OCO2 GMTJIWUFFXGFHH-WPAOEJHSSA-N 0.000 description 1
- RKSBJQZDPAGEQW-WVGOSAFVSA-N saurufurin C Natural products C1([C@H]2O[C@H]([C@@H]([C@@H]2C)C)C=2C=C(C=3OCOC=3C=2)OC)=CC(OC)=C(OC)C(OC)=C1 RKSBJQZDPAGEQW-WVGOSAFVSA-N 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000011272 standard treatment Methods 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
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Abstract
The inventionDiscloses a compound extracted from saururus chinensis and application thereof in preventing and treating nasopharyngeal carcinoma, the compound is shown as a formula (I) and a formula (II),
the compound extracted from the saururus chinensis baill provided by the invention has an obvious effect on nasopharyngeal darcinoma, and the evaluation of the drug effect of the compound shows that the compound obviously inhibits the proliferation of nasopharyngeal darcinoma cells, obviously promotes the apoptosis of cancer cells, has an obvious inhibiting effect on the migration and invasion of the nasopharyngeal darcinoma cells, and can be applied to the preparation of drugs for treating the nasopharyngeal darcinoma. Has good application forward and popularization value.
Description
Technical Field
The invention relates to the technical field of nasopharyngeal carcinoma treatment, and more particularly relates to a compound extracted from saururus chinensis and application thereof in treatment of nasopharyngeal carcinoma.
Background
Nasopharyngeal carcinoma (NPC) is a head and neck malignancy that occurs in the Nasopharyngeal epithelium with high metastasis and high recurrence. The onset of nasopharyngeal carcinoma has regional characteristics, the incidence rate is very low in the world but is extremely high in China, particularly in the Guangdong area in south China, and therefore the nasopharyngeal carcinoma is also called Guangdong carcinoma.
Nasopharyngeal carcinoma is a diverse group of pathogenesis factors, including genetic susceptibility, Epstein-Barr virus (EBV) infection, and a range of environmental risk factors, including smoking and overeating of cured foods. The world health organization histologically classified nasopharyngeal carcinoma into three types, (1) keratinized squamous cell carcinoma (WHO i); (2) differentiated squamous cell carcinoma (WHO II); or undifferentiated squamous cell carcinoma (WHO III); (3) basic squamous cell carcinoma; keratinized squamous cell carcinoma is the most common histological type (WHO i) in the european and american areas; in endemic areas such as china, undifferentiated non-keratinized squamous cell carcinoma (WHO iii) is the most prominent histological type and is closely associated with EBV infection. The current standard treatment scheme of nasopharyngeal carcinoma is the combination of radiotherapy and chemotherapy, the prior treatment usually comprises the emphasis of radiotherapy and cisplatin combination treatment, but about 20 percent of patients are treated by multi-drug chemotherapy due to radiotherapy resistance, and the common chemotherapeutic drugs such as cisplatin and the like have great toxic and side effects, so that the tolerance of the patients is low, and the completion of the treatment is influenced. Therefore, the search for a high-efficiency and low-toxicity anti-nasopharyngeal cancer drug which is easily tolerated by patients is urgently needed clinically.
Disclosure of Invention
The invention aims to overcome the defect that the prior art is lack of a high-efficiency low-toxicity anti-nasopharyngeal-cancer medicament which is easily tolerated by a patient, and provides a compound extracted from saururus chinensis and application thereof in preventing and treating nasopharyngeal cancer.
The first purpose of the invention is to provide a compound extracted from saururus chinensis.
The second purpose of the invention is to provide another compound extracted from saururus chinensis.
The third purpose of the invention is to provide the application of the compound in preparing medicines for treating and/or preventing nasopharyngeal carcinoma.
The fourth purpose of the invention is to provide a medicine for treating and/or preventing nasopharyngeal carcinoma.
In order to achieve the purpose, the invention is realized by the following technical scheme:
the invention claims a compound extracted from saururus chinensis which is shown as a formula (I),
a compound extracted from Saururi herba, wherein the compound is represented by formula (II),
the application of the compound in preparing the medicine for treating and/or preventing nasopharyngeal carcinoma also belongs to the protection scope of the invention.
Preferably, the treatment and/or prevention of nasopharyngeal carcinoma is one or more of inhibiting proliferation of nasopharyngeal carcinoma cells, promoting apoptosis of nasopharyngeal carcinoma cells, or inhibiting migratory invasion of nasopharyngeal carcinoma cells.
The invention also claims a medicine for treating and/or preventing nasopharyngeal carcinoma, which contains the compound.
Preferably, the composition further comprises a pharmaceutically acceptable carrier or auxiliary material.
Compared with the prior art, the invention has the following beneficial effects:
the compound extracted from the saururus chinensis provided by the invention has obvious effect on nasopharyngeal carcinomaThe compound of formula (I) has anti-nasopharyngeal cancer activity IC50To 0.76. mu.M. The evaluation of the drug effect of the compound shows that the compound obviously inhibits the proliferation of nasopharyngeal carcinoma cells, obviously promotes the apoptosis of cancer cells, has obvious inhibiting effect on the migration and invasion of the nasopharyngeal carcinoma cells, and can be applied to the preparation of drugs for treating the nasopharyngeal carcinoma. Has good application forward and popularization value.
Drawings
FIG. 1 is a graph showing the cytotoxicity assay results of compound 13 against four nasopharyngeal carcinoma cells, and the time-and dose-dependent inhibitory activity of compound 13 against the growth of 4 nasopharyngeal carcinoma cells.
FIG. 2 is a graph showing the results of compound 13 inhibiting the clonogenic process of four nasopharyngeal carcinoma cells, and compound 13 inhibits the clonogenic process of 4 nasopharyngeal carcinoma cells.
FIG. 3 is a graph showing the effect of Compound 13 on the HONE1 cell cycle, with Compound 13 inhibiting cell growth and arresting cells in S phase.
Fig. 4 is a graph showing the effect of compound 13 on the apoptosis of HONE1, with compound 13 promoting apoptosis.
Fig. 5 is a graph showing the effect of compound 13 on the migration and invasion of HONE1 cells, and compound 13 inhibits the migration and invasion of cells.
Detailed Description
The invention is described in further detail below with reference to the drawings and specific examples, which are provided for illustration only and are not intended to limit the scope of the invention. The test methods used in the following examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are, unless otherwise specified, commercially available reagents and materials.
Example 1 isolation and extraction of the Compounds
10.0kg of Saururi herba dried aerial parts are pulverized, soaked in 95% ethanol for four times, the leaching solutions are combined, the ethanol is recovered to be small in volume, the small volume is extracted with ethyl acetate and n-butanol for several times in sequence, the extraction solutions are combined, concentrated and dried to obtain ethyl acetate parts (500g) and n-butanol parts (33 g).
Taking about 500g of ethyl acetate part, dissolving ethyl acetate, adsorbing on 500g (200-300 mesh) silica gel, volatilizing at room temperature, performing silica gel column chromatography (2000g), and eluting by adopting a petroleum ether-ethyl acetate system to obtain 10 components, namely Fr.1-Fr.10. Fr.2 by silica gel column chromatography, petroleum ether-acetone system elution, preparing liquid phase and thin layer to obtain compounds 8(1.59g) and 4(130mg), Fr.3 by silica gel column chromatography, petroleum ether-acetone system elution, preparing liquid phase, preparing thin layer and gel column separation to obtain compound 5(44mg), compound 7 (saururus) (6 g); fr.4 performing silica gel column chromatography, eluting with petroleum ether-acetone system, preparing thin layer, and separating by silica gel column chromatography repeatedly to obtain compounds 6(25mg), 9(577mg), 10(5mg), and 11(40 mg); fr.5 performing silica gel column chromatography, eluting with dichloromethane, preparing liquid phase, preparing thin layer, and separating with gel column to obtain compounds 3(271mg) and 1(38 mg); fr.8 gel column chromatography, methanol elution, preparation of liquid phase, preparation of thin layer, repeated separation of gel and silica gel column to give compounds 13(412mg), 17(1.0g), 16(109mg), 2(8mg), 12(45mg), 15(36mg) and 14(58 mg).
The identification data for the chemical composition of 17 monomers are as follows:
compound 1
Colorless oil, C22H26O6。EI-MS m/z:369[M+H-H2O]+，399。1H-NMR(400MHz,CDCl3)δ:6.98(1H,d,J＝1.84Hz,H-6′),6.96(1H,dd,J＝1.84,8.08Hz,H-2′),6.86(1H,d,J＝8.08Hz,H-3′),6.65(1H,d,J＝1.16Hz,H-6),6.62(1H,J＝1.16Hz,H-2),5.96(2H,s,-OCH2O-),4.49(1H,d,J＝6.64Hz,H-7),4.46(1H,d,J＝6.32Hz,H-7′),3.89(3H,s,-OMe),3.89(3H,s,-OMe),3.88(3H,s,-OMe),2.29(2H,m,H-8、8′),1.04(3H,d,J＝6.76Hz,9-Me),1.02(3H,d,J＝6.72Hz,9′-Me)。13C-NMR(100MHz,CDCl3)δ:137.0(s,C-1),106.1(d,C-2),143.4(s,C-3),134.5(s,C-4),148.9(s,C-5),100.3(d,C-6),87.4(d,C-7),44.6(d,C-8),13.0(q,C-9),134.5(s,C-1′),118.6(d,C-2′),110.9(d,C-3′),148.5(s,C-4′),148.9(s,C-5′),109.7(d,C-6′),87.2(d,C-7′),44.2(d,C-8′),12.8(q,C-9′),55.8(q,-OMe),55.9(q,-OMe),56.6(q,-OMe),101.4(t,-OCH2O-)。
Colorless oil, C22H26O5。EI-MS m/z:387[M+H]+。1H-NMR(400MHz,CDCl3)δ:6.99(1H,s,H-2),6.96(1H,d,J＝8.14Hz,H-6),6.86(1H,d,J＝8.14Hz,H-5),6.65(1H,s,H-2′),6.62(1H,s,H-6′),5.95(2H,s,-OCH2O-),4.49(1H,d,J＝6.48Hz,H-7),4.46(1H,d,J＝6.37Hz,H-7′),3.89(3H,s,-OMe),3.89(3H,s,-OMe),3.88(3H,s,-OMe),2.28(2H,m,H-8、8′),1.06(3H,d,J＝7.70Hz,9-Me),1.02(3H,d,J＝7.12Hz,9′-Me)。13C-NMR(100MHz,CDCl3)δ:134.5(s,C-1),109.8(d,C-2),148.5(s,C-3),149.0(s,C-4),111.0(d,C-5),118.6(d,C-6),87.4(d,C-7),44.6(d,C-8),13.0(q,C-9),134.6(s,C-1′),106.2(d,C-2′),143.4(s,C-3′),137.1(s,C-4′),148.9(s,C-5′),100.3(d,C-6′),87.2(d,C-7′),44.2(d,C-8′),12.8(q,C-9′),55.8(q,-OMe),55.9(q,-OMe),56.6(q,-OMe),101.3(t,-OCH2O-)。
Compound 3(Saurucinol I)
Colorless oil, C23H28O7。EI-MS m/z:417[M+H]+，399。1H-NMR(400MHz,CDCl3)δ:6.66(3H,s,H-2、6、6′),6.62(1H,s,H-2′),5.95(2H,s,-OCH2O-),4.49(1H,d,J＝6.52Hz,H-7),4.47(1H,d,J＝6.68Hz,H-7′),3.92(3H,s,4-OMe),3.86(3H,s,3-OMe),3.86(3H,s,5-OMe),3.84(3H,s,4′-OMe),2.29(2H,m,H-8、8′),1.06(3H,d,J＝6.44Hz,9-Me),1.03(3H,d,J＝6.52Hz,9′-Me)。13C-NMR(100MHz,CDCl3)δ:136.8(s,C-1),103.3(d,C-2),153.2(s,C-3),137.4(s,C-4),153.2(s,C-5),103.3(d,C-6),87.5(d,C-7),44.4(d,C-8),13.1(q,C-9),134.6(s,C-1′),106.4(d,C-2′),143.4(s,C-3′),137.8(s,C-4′),149.0(s,C-5′),100.4(d,C-6′),87.3(d,C-7′),44.3(d,C-8′),12.8(q,C-9′),56.1(q,3-OMe),60.8(q,4-OMe),56.1(q,5-OMe),56.6(q,5′-OMe),101.4(t,-OCH2O-)。
Compound 4((-) -Zuonin)
Colorless massive crystals (chloroform), C20H20O5。EI-MS m/z:341[M+H]+,323[M+H-H2O]+。1H-NMR(400MHz,CDCl3)δ:6.77-6.93(6H,m,H-2、2′、5、5′、6、6′),5.94(4H,s,2×-OCH2O-),5.40(1H,d,J＝9.48Hz),4.62(1H,d,J＝4.47Hz),2.41(2H,m,H-8、8′),1.00(3H,d,J＝6.98Hz),0.63(3H,d,J＝6.53Hz)。13C-NMR(100MHz,CDCl3)δ:134.9,134.4(s,C-1,1′),119.4,118.9(d,C-2,2′),147.7,147.3(s,C-3,3′),146.8,146.2(s,C-4,4′),106.6,106.3(d,C-5,5′),107.8,107.8(d,C-6,6′),100.8,100.7(t,2×-OCH2O-),85.6,84.6(d,C-7,7′),47.4,43.3(d,C-8,8′),11.7,9.3(q,C-9,9′)。
Compound 5
A colorless powder, C20H20O6。EI-MS m/z:357[M+H]+。1H-NMR(400MHz,CDCl3)δ:6.74(1H,s,H-6),6.54(1H,s,H-3),5.95(1H,s,Ar.OCH2O),5.93(1H,s,Ar.OCH2O),5.76(1H,s,Al.OCH2O),5.62(1H,s,H-3′),5.53(1H,s,Al.OCH2O),2.74(1H,m,H-1′),2.33(1H,td,J＝13.7,2.7Hz,H-7′eq),2.24(1H,dd,J＝12.5,2.7Hz,H-6′),2.15(1H,d,J＝9.7Hz,H-7),1.48(1H,m,H-8′),1.45(1H,m,H-8),1.15(3H,d,J＝5.0Hz,H-9),1.07(1H,m,H-7′ax),1.07(3H,d,J＝6.1Hz,H-9′)。13C-NMR(100MHz,CDCl3)δ:126.32(d,C-1),146.16(s,C-2),101.69(d,C-3),144.02(s,C-4),146.21(s,C-5),105.12(d,C-6),41.87(d,C-7),36.88(d,C-8),18.63(q,C-9),45.41(d,C-1′),198.02(s,C-2′),101.24(d,C-3′),164.09(s,C-4′),105.31(s,C-5′),42.67(d,C-6′),32.36(t,C-7′),34.06(d,C-8′),20.14(q,C-9′),101.35(t,C-Ar.OCH2O),97.68(t,C-Al.OCH2O)。
Compound 6 (1' -Epi-sachhinone)
A colorless powder, C20H20O6。EI-MS m/z:357[M+H]+。1H-NMR(400MHz,CDCl3)δ:6.78(1H,s,H-6),6.36(1H,s,H-3),5.92(1H,s,Ar.OCH2O),5.89(1H,s,Ar.OCH2O),5.62(1H,s,Al.OCH2O),5.61(1H,s,Al.OCH2O),5.58(1H,s,H-3′),3.19(1H,dd,J＝1.96,8.8Hz,H-7),2.88(1H,t,J＝8.9,H-6′),2.66(1H,m,H-1′),2.24(1H,m,H-8),2.04(1H,m,H-7′eq),1.58(1H,m,H-8′),1.22(3H,d,J＝7.52,H-9),1.16(1H,m,H-7′ax),0.70(3H,d,J＝7.08,H-9′)。13C-NMR(100MHz,CDCl3)δ:118.8(d,C-1),144.97(s,C-2),100.31(d,C-3),143.29(s,C-4),146.52(s,C-5),107.34(d,C-6),36.17(d,C-7),35.26(d,C-8),24.54(q,C-9),39.47(d,C-1′),199.43(s,C-2′),99.50(d,C-3′),168.23(s,C-4′),100.74(s,C-5′),42.19(d,C-6′),30.79(t,C-7′),32.76(d,C-8′),23.58(q,C-9′),101.23(t,C-Ar.OCH2O),98.18(t,C-Al.OCH2O)。
Compound 7(Sauchinone)
A colorless powder, C20H20O6。EI-MS m/z:357[M+H]+. The Rf values were the same with the saururus chinensis ketone standard by silica gel TLC using petroleum ether/acetone (9: 1, Rf 0.3) and dichloromethane/n-hexane (4: 1, Rf 0.4) as developing agent. The melting point of the mixture is 224-226 ℃, the melting point of the mixture and the saururus chinensis ketone is not reduced, and the compound is identified as the saururus chinensis ketone.
Compound 8(Licarin B)
White needle crystals (chloroform), C20H20O4。EI-MS m/z:325[M+H]+。1H-NMR(400MHz,CDCl3)δ:6.93(1H,d,J＝1.64Hz,H-2′),6.88(1H,dd,J＝1.64,7.96Hz,H-6′),6.78(1H,d,J＝7.96Hz,H-5′),6.78(1H,s,H-4),6.75(1H,s,H-6),6.36(1H,dd,J＝1.60,15.70Hz,H-8),6.10(1H,dq,J＝6.6,15.70Hz,H-9),5.95(2H,s,-OCH2O-),5.09(1H,d,J＝8.88Hz,H-2),3.89(3H,s,7-OMe),3.41(1H,m,H-3),1.87(3H,dd,J＝1.6,6.6Hz,H-10)。13C-NMR(100MHz,CDCl3)δ:93.4(d,C-2),45.8(d,C-3),133.1(s,C-3a),113.4(d,C-4),132.2(s,C-5),109.4(d,C-6),144.11(s,C-7),147.6(s,C-7a),130.9(d,C-8),123.4(d,C-9),18.32(q,C-10),134.4(s,C-1′),106.8(d,C-2′),146.5(s,C-3′),147.9(s,C-4′),108.0(d,C-5′),120.1(d,C-6′),17.9(q,3-Me),101.1(t,-OCH2O-),56.0(q,7-OMe)。
Compound 9(Licarin A)
White crystals (chloroform), C20H22O4。EI-MS m/z:327[M+H]+。1H-NMR(400MHz,CDCl3)δ:6.98(1H,d,J＝1.5Hz,H-2′),6.90(1H,dd,J＝1.5,8.5Hz,H-6′),6.88(1H,d,J＝8.5Hz,H-5′),6.79(1H,s,H-4),6.77(1H,s,H-6),6.36(1H,dd,J＝1.28,15.68Hz,H-8),6.11(1H,dq,J＝6.8,15.68Hz,H-9),5.64(1H,s,4′-OH),5.10(1H,d,J＝9.44Hz,H-2),3.90(3H,s,7-OMe),3.88(3H,s,3′-OMe),3.45(1H,dq,J＝6.8,9.44Hz,H-3),1.88(3H,d,J＝6.8Hz,H-10),1.38(3H,d,J＝6.8Hz,3-Me)。13C-NMR(100MHz,CDCl3)δ:93.8(d,C-2),45.6(d,C-3),133.24(s,C-3a),113.28(d,C-4),132.1(s,C-5),109.2(d,C-6),144.1(s,C-7),146.5(s,C-7a),130.9(d,C-8),123.4(d,C-9),18.4(q,C-10),132.2(s,C-1′),108.9(d,C-2′),146.6(s,C-3′),145.8(s,C-4′),114.0(d,C-5′),119.9(d,C-6′),17.5(q,3-Me),56.0(q,7-OMe),55.9(q,3′-OMe)。
Compound 10(Eupomatenoid-7)
White powder, C20H20O4。EI-MS m/z:325[M+H]+。1H-NMR(400MHz,CDCl3)δ:7.32(1H,d,J＝1.68Hz,H-2′),7.29(1H,dd,J＝1.68,8.24Hz,H-6′),7.04(1H,d,J＝1.2Hz,H-4),7.00(1H,d,J＝8.24Hz,H-5′),6.83(1H,d,J＝1.2Hz,H-6),6.50(1H,dd,J＝1.60,15.80Hz,H-8),6.22(1H,dq,J＝6.5,15.80Hz,H-9),4.04(3H,s,7-OMe),3.99(3H,s,3′-OMe),2.41(3H,s,3-Me),1.91(3H,dd,J＝1.6,6.5Hz,H-10)。13C-NMR(100MHz,CDCl3)δ:151.5(s,C-2),110.2(s,C-3),142.1(s,C-3a),109.2(d,C-4),133.1(s,C-5),109.5(d,C-6),133.7(s,C-7),146.6(s,C-7a),131.5(d,C-8),124.4(d,C-9),18.41(q,C-10),123.7(s,C-1′),104.5(d,C-2′),144.9(s,C-3′),145.8(s,C-4′),114.5(d,C-5′),120.7(d,C-6′),9.59(q,3-Me),56.14(q,7-OMe),56.10(q,3′-OMe)。
Compound 11((2R,3R) -2, 3-hidro-2- (4-hydroxy-3-methoxyphenyl) -7-methoxy-3-tolylfluran-5-aldehydel)
White amorphous powder (MeOH), C18H18O5。EI-MS m/z:315[M+H]+。1H-NMR(400MHz,CDCl3)δ:9.84(1H,s,H-8),7.37(1H,s,H-4),7.34(1H,s,H-6),6.90-6.93(3H,m,H-2′,5′,6′),5.76(1H,s,4′-OH),5.24(1H,d,J＝9.24Hz,H-2),3.94(3H,s,3′-OMe),3.88(3H,s,7-OMe),3.55(1H,m,H-3),1.44(3H,d,J＝6.8Hz,3-Me)。13C-NMR(100MHz,CDCl3)δ:95.0(d,C-2),44.8(d,C-3),133.6(s,C-3a),120.0(d,C-4),131.0(s,C-5),111.7(d,C-6),146.7(s,C-7),153.2(s,C-7a),190.6(s,C-8),131.4(s,C-1′),108.8(d,C-2′),146.1(s,C-3′),144.9(s,C-4′),114.3(d,C-5′),120.0(d,C-6′),17.7(q,3-Me),56.0(q,7-OMe),55.9(q,3′-OMe)。
Compound 12, a compound of formula (II)
White powder, C30H36O8。EI-MS m/z:507[M-H2O+H]+,489。1H-NMR(400MHz,CDCl3)δ:6.97(1H,d,J＝8.16Hz,H-5′),6.92(1H,m,H-6″),6.90(1H,d,J＝1.7Hz,H-2′),6.89(1H,m,H-5),6.86(2H,m,H-2″,5″),6.83(1H,d,J＝1.7Hz,H-2),6.82(1H,m,H-6′),6.76(1H,m,H-6),5.76(1H,brs,-OH),5.44(1H,d,J＝5.96Hz,H-7),5.43(1H,d,J＝5.96Hz,H-7′),4.62(1H,d,J＝8.28Hz,H-7″),4.12(1H,m,H-8″),3.86-3.91(9H,s,3,3′,3″-OMe),2.27(2H,m,H-8、8′),1.15(3H,d,J＝6.20Hz,9″-Me),0.70(3H,d,J＝6.16Hz,9-Me),0.69(3H,d,J＝5.36Hz,9′-Me)。13C-NMR(100MHz,CDCl3)δ:133.2(s,C-1),108.9(d,C-2),146.4(s,C-3),144.4(s,C-4),114.1(d,C-5),119.0(d,C-6),83.5(d,C-7),44.1(d,C-8),14.8(q,C-9),136.5(s,C-1′),110.1(d,C-2′),150.5(s,C-3′),146.3(s,C-4′),118.5(d,C-5′),118.7(d,C-6′),83.3(d,C-7′),44.0(d,C-8′),14.7(q,C-9′),131.9(s,C-1″),120.6(d,C-2″),146.6(s,C-3″),145.4(s,C-4″),113.9(d,C-5″),109.4(d,C-6″),78.4(d,C-7″),83.9(d,C-8″),16.9(q,C-9″),55.8-55.9(q,3,3′,3″-OMe)。
Compound 13(Saucerneol), a compound of formula (I)
A colorless powder, C31H38O8。EI-MS m/z:556[M+NH4]+,503。13C-NMR(100MHz,CDCl3)δ:133.1(s,C-1),108.9(d,C-2),146.3(s,C-3),146.3(s,C-4),110.1(d,C-5),119.0(d,C-6),83.5(d,C-7),44.1(d,C-8),14.7(q,C-9),136.5(s,C-1′),110.9(d,C-2′),150.5(s,C-3′),144.5(s,C-4′),118.6(d,C-5′),118.7(d,C-6′),83.3(d,C-7′),44.0(d,C-8′),14.7(q,C-9′),132.5(s,C-1″),110.2(d,C-2″),149.0(s,C-3″),148.8(s,C-4″),113.9(d,C-5″),119.9(d,C-6″),78.3(d,C-7″),83.8(d,C-8″),16.9(q,C-9″),55.8-55.9(q,3,3′,3″,4″-OMe)。
Compound 14(Saucerneol methyl ether)
A colorless powder, C32H40O8。EI-MS m/z:570[M+NH4]+,517。1H-NMR(400MHz,CDCl3)δ:6.81-6.98(9H,m,H-2,5,6,2′,5′,6′,2″,5″,6″),5.44(1H,d,J＝5.96Hz,H-7),5.44(1H,d,J＝5.96Hz,H-7′),4.63(1H,d,J＝8.24Hz,H-7″),4.12(1H,m,H-8″),3.85-3.91(15H,s,3,4,3′,3″,4″-OMe),2.27(2H,m,H-8,8′),1.15(3H,d,J＝6.20Hz,H-9″),0.69(3H,d,J＝5.76Hz,H-9),0.68(3H,d,J＝5.76Hz,H-9′)。13C-NMR(100MHz,CDCl3)δ:133.8(s,C-1),109.5(d,C-2),147.8(s,C-3),148.5(s,C-4),110.0(d,C-5),118.6(d,C-6),83.4(d,C-7),44.0(d,C-8),14.7(q,C-9),136.5(s,C-1′),110.1(d,C-2′),150.4(s,C-3′),146.3(s,C-4′),118.3(d,C-5′),118.5(d,C-6′),83.3(d,C-7′),44.0(d,C-8′),14.7(q,C-9′),132.5(s,C-1″),110.7(d,C-2″),148.9(s,C-3″),148.7(s,C-4″),110.8(d,C-5″),119.9(d,C-6″),78.2(d,C-7″),83.9(d,C-8″),16.9(q,C-9″),55.7-55.8(q,3,4,3′,3″,4″-OMe)。
Compound 15((-) - (7 "R, 8" R) -Saucerneol J)
White powder, C30H36O8。EI-MS m/z:542[M+NH4]+,489。1H-NMR(400MHz,CDCl3)δ:7.02(1H,d,J＝1.7Hz,H-2′),6.99(1H,m,H-6″),6.97(1H,m,H-6′),6.93(1H,m,H-5″),6.92(1H,m,H-2″),6.92(1H,m,H-5),6.92(1H,d,J＝1.7Hz,H-2),6.86(1H,m,H-5′),6.86(1H,m,H-6),5.71(1H,brs,-OH),5.14(1H,d,J＝8.64Hz,H-7),4.62(1H,d,J＝8.32Hz,H-7″),4.41(1H,d,J＝9.32Hz,H-7′),4.11(1H,m,H-8″),3.87-3.90(9H,s,3,3′,3″-OMe),2.25(1H,m,H-8′),1.80(1H,m,H-8),1.15(3H,d,J＝6.24Hz,9″-Me),1.06(3H,d,J＝6.52Hz,9′-Me),0.68(3H,d,J＝7.0Hz,9-Me)。13C-NMR(100MHz,CDCl3)δ:136.3(s,C-1),110.9(d,C-2),150.4(s,C-3),145.4(s,C-4),118.4(d,C-5),119.5(d,C-6),82.9(d,C-7),45.9(d,C-8),15.0(q,C-9),132.6(s,C-1′),109.4(d,C-2′),146.5(s,C-3′),146.6(s,C-4′),114.1(d,C-5′),119.2(d,C-6′),87.4(d,C-7′),47.6(d,C-8′),15.0(q,C-9′),131.9(s,C-1″),109.4(d,C-2″),146.5(s,C-3″),145.2(s,C-4″),114.2(d,C-5″),120.6(d,C-6″),78.4(d,C-7″),83.9(d,C-8″),16.9(q,C-9″),55.7-55.9(q,3,3′,3″-OMe)。
Compound 16 (4-O-Demethylmassantin B)
A colorless powder, C40H46O11。EI-MS m/z:679[M+Na-OCH2O]+,649[M+Na--OCH2O--OCH2]+。1H-NMR(400MHz,CDCl3)δ:6.76-6.99(12H,m,H-2,5,6,2′,5′,6′,2″,5″,6″,2″′,5″′,6″′),5.94(2H,s,-OCH2O-),5.64(1H,brs,-OH),5.46(2H,d,J＝5.52Hz,H-7′,7″),4.62(1H,d,J＝8.04Hz,H-7),4.61(1H,d,J＝8.04Hz,H-7″′),4.10(2H,m,H-8,8″′),3.89-3.92(9H,s,3,3′,3″-OMe),2.29(2H,m,H-8′,8″),1.16(3H,d,J＝4.92Hz,H-9″′),1.15(3H,d,J＝5.0Hz,H-9),0.72(6H,d,J＝5.68Hz,H-9′,9″)。13C-NMR(100MHz,CDCl3)δ:150.6(s,C-4′,4″),147.7(s,C-3″′),147.4(s,C-4″′),146.6(s,C-3),146.5(s,C-3″),146.3(s,C-3′),145.5(s,C-4),136.6(s,C-1″),136.5(s,C-1′),134.0(s,C-1″′),132.0(s,C-1),121.0(d,C-6),120.7(d,C-6″′),118.9(d,C-5′),118.7(d,C-5″,6″,6),114.1(d,C-5),110.2(d,C-2″,2′),109.5(d,C-2),108.1(d,C-5″′),107.5(d,C-2″′),101.0(t,-OCH2O-),84.1(d,C-8″′),83.9(d,C-8),83.4(d,C-7′,7″),78.5(d,C-7″′),78.4(d,C-7),55.94(q,-OCH3),55.87(q,-OCH3×2),44.2(d,C-8′,8″),17.0(q,C-9″′),16.9(q,C-9),14.8(q,C-9′,9″)。
Compound 17(Manassantin B)
A colorless powder, C41H48O11。EI-MS m/z:735[M+H+H2O]+,682,663。1H-NMR(400MHz,CDCl3)δ:6.72-6.97(12H,m,H-2,5,6,2′,5′,6′,2″,5″,6″,2″′,5″′,6″′),5.88(2H,s,-OCH2O-),5.43(2H,d,J＝5.68Hz,H-7,7′),4.63(1H,d,J＝8.04Hz,H-7″),4.59(1H,d,J＝8.12Hz,H-7″′),4.12(2H,m,H-8″,8″′),3.83-3.88(12H,s,3,3′,3″,4″-OMe),2.27(2H,m,H-8,8′),1.13(3H,d,J＝6.60Hz,H-9″),1.12(3H,d,J＝6.60Hz,H-9″′),0.69(6H,d,J＝5.64Hz,H-9,9′)。13C-NMR(100MHz,CDCl3)δ:150.3(s,C-4,4′),148.8(s,C-4″′),148.6(s,C-4″),147.5(s,C-3″′),147.1(s,C-3″),146.2(s,C-3′),146.1(s,C-3),136.3(s,C-1′),136.2(s,C-1),133.9(s,C-1″′),132.6(s,C-1″),120.8(d,C-6″′),119.7(d,C-6″),118.5(d,C-5″,5″′,6),118.3(d,C-6′),110.7(d,C-2″′),110.0(d,C-2,2′,2″),107.8(d,C-5),107.3(d,C-5′),100.8(t,-OCH2O-),83.4(d,C-8″,8″′),83.2(d,C-7,7′),78.0(d,C-7″,7″′),55.6(q,-OCH3×4),43.9(d,C-8,8′),16.7(q,C-9″),16.6(q,C-9″′),14.6(q,C-9,9′)。
The structural formulas of compounds 1-17 are shown below:
EXAMPLE 2 determination of anti-nasopharyngeal carcinoma Activity of Compounds
First, culture of cells
(1) Cell resuscitation
Frozen CNE1, CNE2, HONE1 and SUNE1 cells are taken out of a liquid nitrogen tank, immediately placed into a water bath at 37 ℃ and shaken within 1min to be quickly dissolved, the dissolved cells are transferred into a centrifuge tube filled with RPMI 1640 culture medium containing 5ml of 10% FBS, centrifuged at 800rpm for 3min, the culture medium is discarded, a proper amount of culture medium is added, the mixture is evenly blown and beaten, the mixture is transferred into a culture dish and cultured in a culture box with the culture condition of 37 ℃ and 5% CO2, the liquid is changed every other day, and the passage is carried out once every three days.
(2) Cell passage
When the cells were observed to grow to about 80% confluence, the culture solution was discarded, the serum and floating cells were washed off with PBS, and then digested with 1mL of 0.25% EDTA-containing trypsin for about 3min, it was observed that when 80% of the cells became round, the digestion was stopped by immediately adding 10% FBS-containing medium, the cells were gently blown with a pipette and transferred to a 5mL centrifuge tube, centrifuged at 800rpm for 3min, and the supernatant was carefully discarded. Adding a proper amount of culture medium, gently blowing and beating to uniformly mix the cells, and carrying out passage at a ratio of 1: 3.
(3) Cell cryopreservation
Digesting 80% confluent cells in logarithmic phase growth phase with 0.25% pancreatin containing EDTA, centrifuging at 800rpm for 3min, collecting cells, discarding supernatant, blowing with frozen stock solution containing 10% DMSO and 90% serum, mixing, counting cells, and adjusting density to 1 × 106Transferring the cells/ml to a sterile freezing tube, sealing and marking, putting into a programmed gradient cooling box, freezing and storing overnight at-80 ℃, and then transferring into a liquid nitrogen tank.
Second, MTT experiment method for measuring cell inhibition rate
1. Experimental methods
The CNE1, CNE2, SUNE1, and HONE1 cells in good condition were discarded from the original medium, washed twice with PBS, trypsinized cells added with 1ml of 0.25% EDTA, centrifuged at 800rpm for 3min to collect cell pellets, resuspended in complete medium, counted using a cell counting plate, seeded at a seeding density of 3000 cells/well in a 96-well plate (100. mu.L/well), and cultured in a 5% CO2 incubator at 37 ℃ for 24 hours. After the cells are attached to the wall, the original culture medium is discarded, compounds with different concentrations are used for replacing the original culture medium, the blank is the culture medium without the cells, and the control is the cells without the medicines. Each well is 100. mu.L, and each set is provided with three multiple wells. The 96-well plate was placed in an incubator and incubated for 24 hours, 48 hours, and 72 hours, respectively. After completion of the incubation, 20. mu.L (5mg/mL) of MTT solution was added, the incubation was continued for 4 hours, followed by carefully aspirating the supernatant, adding 150. mu.L of DMSO, shaking for 10 minutes, and then measuring the OD value thereof at 492nm using a microplate reader. The inhibition rate calculation formula is as follows: 1- (drug addition OD-blank OD)/(control OD-blank OD) × 100%.
2. Results of the experiment
The inhibition rates of 17 compounds are shown in table 1.
TABLE 1 cellular Activity of Saururi herba aerial part 17 monomer Compound on four nasopharyngeal carcinomas
aThe inhibition rate of the compound on four nasopharyngeal carcinoma cells under the concentration of 10 mu M;bthe compound has no inhibitory activity at a concentration of 10 μ M;ccisplatin was used as a positive control.
As shown in Table 1, the compounds represented by the formula (II) (compound 12) and the compound represented by the formula (I) (compound 13) each had a very good inhibitory effect on various nasopharyngeal cancer cell lines, and the inhibitory effect was similar to that of cisplatin.
Determination of the triple and half inhibitory concentrations
1. Experimental methods
Half maximal inhibitory concentrations of compound 12 and compound 13 were determined on 4 nasopharyngeal carcinoma cells (CNE2, CNE1, SUNE1, HONE1) using MTT reagent, respectively. Cells in logarithmic growth phase at 3.0X 104The cells were seeded in 96-well plates at a density of 100. mu.L/well and allowed to adhere to the walls by culturing overnight in a 5% CO2 incubator at 37 ℃. Cells were then treated with different concentrations of compound 12 and compound 13 for 24, 48, 72h and tested with MTT reagent. The method comprises the following specific steps: mu.L of the prepared MTT solution (5mg/ml) was added to each well and incubation was continued at 37 ℃ for 4 h. Formazan crystals were dissolved with 150. mu.L DMSO on a shaker for 15 minutes, and then absorbance was measured at 492nm using a microplate reader. The blank group contained medium only, and the control group was a group of cells cultured normally. The proliferation rate is calculated by the formula: the proliferation rate was (experimental OD-blank OD)/(control OD-blank OD) × 100. The experiment is repeated for 3 times, the GraphPad software is used for drawing a dose-dependent inhibition curve of the compound 12 and the compound 13 on four nasopharyngeal carcinoma cell lines, and the half Inhibition Concentration (IC) of the compound 12 and the compound 13 on the four nasopharyngeal carcinoma cell lines is obtained50Values) to design the optimal drug concentration for the following experiments.
2. Results of the experiment
TABLE 2 inhibitory Activity of Compounds 13 and 12 on nasopharyngeal carcinoma cell proliferation
As shown in Table 2, the semi-inhibitory concentrations of Compound 12 and Compound 13 against various nasopharyngeal carcinoma cell lines were low, similar to that of cisplatin. And the semi-inhibitory concentration of the compound 13 is lower, the activity on HONE1 nasopharyngeal carcinoma cells is 10 times stronger than that of the positive drug cisplatin, and the compound has good inhibitory effect on nasopharyngeal carcinoma.
EXAMPLE 3 cytotoxicity assay of Compound 13 against four nasopharyngeal carcinoma cells
1. Experimental methods
The well-conditioned CNE1, CNE2, SUNE1 and HONE1 were seeded in a 96-well plate (100. mu.L/well) at a seeding density of 3000 cells/well, and cultured in a 5% CO2 incubator at 37 ℃ for 24 hours. After the cells are attached to the wall, the original culture medium is discarded, and the compound 13 with different concentrations is used for replacing the original culture medium, the blank is the culture medium without the cells, and the control is the cells without the medicine. Each well is 100. mu.L, and each set is provided with three multiple wells. The 96-well plate was placed in an incubator and incubated for 24 hours, 48 hours, and 72 hours, respectively. After completion of the incubation, 20. mu.L (5mg/mL) of MTT solution was added, the incubation was continued for 4 hours, followed by carefully aspirating the supernatant, adding 150. mu.L of DMSO, shaking for 10 minutes, and then measuring the OD value thereof at 492nm using a microplate reader. The survival rate calculation formula is as follows: (the drug adding component OD-blank group OD)/(the control group OD-blank group OD) is 100%.
2. Results of the experiment
As shown in FIG. 1, Compound 13 significantly reduced the survival of four nasopharyngeal carcinoma cells (CNE1, CNE2, HONE1, SUNE1) and showed significant time-concentration dependence. In particular, compound 13 was most effective in cytotoxic effect against HONE1, and after 48 hours, 0.1. mu.M of compound 13 resulted in a survival rate of HONE1 cells of less than 30. The experimental result shows that the compound 13 has more obvious cytotoxicity on HONE1 cells, and HONE1 is selected in subsequent experiments to further study the proliferation inhibition activity of the compound.
EXAMPLE 4 Compound 13 inhibits clonogenic formation of four nasopharyngeal carcinoma cells
1. Experimental methods
After the HONE1 cells in the logarithmic growth phase are digested by trypsin, 500 cells are inoculated in a 6-well plate per well, compound 13(0, 1, 3 and 10 mu M) with different concentrations is added for incubation for 7 days, the medicine is changed once every two days, after the culture is finished, the cells are fixed by 4% paraformaldehyde, then 1% crystal violet is used for staining for 30 minutes, the mixture is air-dried at room temperature, the colony number is counted, and every 50 cells are taken as a colony.
2. Results of the experiment
As shown in FIG. 2, Compound 13 significantly inhibited the formation of four clones of nasopharyngeal carcinoma cells at 1. mu.M, with the reduction in the number of clones formed by HONE1 cells being most significant for Compound 13, indicating that it significantly inhibited its proliferative activity, in agreement with the above cytotoxicity assay.
EXAMPLE 5 Effect of Compound 13 on cell cycle
1. Experimental methods
Flow cytometry detection of cell cycle, 4X 105HONE1 cell species/well are cultured in 6-well plates for 24h, Compound 13 is treated with HONE1 cells at different concentrations (0, 1, 3, 10. mu.M) for 24h, followed by trypsinization, cell collection by centrifugation, fixation with frozen 70% ethanol overnight at 4 ℃, followed by washing with cold PBS buffer, followed by photophobic staining with a cell cycle and apoptosis kit at 37 ℃ for 30 min the treated samples are analyzed with a Beckmann flow cytometer and 15000 cells are collected from each sample the results are analyzed in Modfit software the cycle-related protein assay is performed using the Western Blot protocol by treating 24h HONE1 cells with different concentrations of Compound 13, first lysing the cells on ice with RIPA lysate, then recovering the total protein with a cell scraper, determining the total protein content with a BCA protein quantification kit, denaturing the proteins are separated by electrophoresis with 12% SDS-PAGE gel, cutting the gel, electroporating the proteins onto PVDF membrane, followed by incubating the electroporated PVDF membrane with a blocking solution (5% CDK) for 2h, adding corresponding anti-CDK antibodies (CDK) for 2 min at room temperature, adding corresponding antibodies 2 Bucht 1) to the corresponding antibodies (TBST 3-7 min, adding BUT antibody, incubating with TBST 54 min, 3-N antibody, incubating with a corresponding cycle-wash-dry cellh, washing the membrane with TBST for 3 times, 10 minutes each time; and finally, adding chemiluminescence liquid to develop in a developing instrument, and finally carrying out gray level analysis on the protein band by using Image J software.
2. Results of the experiment
As shown in FIGS. 3A and 3B, compound 13 was able to block the cell cycle of HONE1 nasopharyngeal carcinoma cells in S phase and was concentration dependent. To further validate the effect of compound blockade at S phase, immunoblotting was used to detect expression of key proteins at the S phase checkpoint. The results in fig. 3C show that after compound 13 treated HONE1 cells for 24h, the protein expression levels of cyclin d1 and cyclin dependent kinase CDK2 were all decreased, consistent with the results of the S-phase block of the cell cycle.
EXAMPLE 6 Effect of Compound 13 on apoptosis
1. Experimental methods
Apoptosis experiments were performed using Annexin V-FITC/PI kit for 24h, trypsinizing the collected cells with 0.25% EDTA in pancreatin, washing with ice PBS to remove residual pancreatin, resuspending the cells in 100uL of buffer in the kit, adding 5 uL of Annexin V-FITC and 10uLPI solutions, incubating for 15 min at room temperature, detecting the fluorescent signals of apoptosis in an up-flow cytometer within 1h, collecting 15000 cells in each sample, analyzing in FlowJoVX software, apoptotic protein detection using Western Blot assay method for 24h of HONE1 cells treated with different concentrations of compound 13, first lysing on ice with RIPA lysate, then collecting the cells, using scraping protein quantification kit to determine the total protein content, denaturing the proteins, separating with 12% SDS-gel, cleaving the proteins, separating with PVDF gel, washing with PVDF gel, performing electrophoresis for 10 min, performing electrophoresis on PVDF gel electrophoresis strips, performing electrophoresis on PVDF gel electrophoresis gel for 10 min, performing electrophoresis on a dry protein blotting, performing electrophoresis on a dry protein assay using a dry cell-gel electrophoresis method, performing a dry cell-wash, performing electrophoresis for 10 min, performing a dry cell-wash test on a dry cell wash test, performing a dry cell wash, performing.
2. Results of the experiment
The results are shown in fig. 4, compound 13 can promote apoptosis of HONE1 nasopharyngeal carcinoma cells, and the apoptosis rate is increased from 6.19% to 58% with the increase of the concentration of compound 13, which has a significant difference compared with the blank group. In order to further verify the apoptosis, Western Blot is used for detecting apoptosis-related proteins, and the result shows that the compound 13 can obviously down-regulate the apoptosis protein Bcl-2 and lead the cleavage substrate PARP of the apoptosis core member caspase (caspase) to show a remarkable down-regulation trend, and meanwhile, the full-length PARP (116KDa) is divided into cleaned PARP (89 KDa). The expression level of cleared PARP appeared at a concentration of 10. mu.M of Compound 13. Cleaved PARP results in inhibited DNA repair and DNA degradation, resulting in compound 13-induced apoptosis of HONE1 cells.
EXAMPLE 7 Effect of Compound 13 on cell transfer
1. Experimental methods
Detection of the inhibitory effect of compound 13 on cell metastasis two methods were used to mutually validate the results of the experiment.
The method comprises the following steps: cell scratch test
The cells of HONE1 in the logarithmic growth phase are inoculated in a 6-well plate, after the cells adhere to the wall, a straight line is smoothly drawn in the center of the six-well plate by using a 100-mu-L gun head, the cells are washed for three times by using PBS (phosphate buffer solution), suspension cells marked at the scratch are removed, 13(0, 1, 3 and 10 mu M) culture media with different concentrations and containing 1% FBS (FBS) are replaced, the cells are placed in an incubator for culture, the migration conditions of the HONE1 cells at 0h, 24h and 48h are observed, and the migration states of the cells at the same position are recorded by photographing. And finally, calculating the blank area of the scratch by using Image J software.
The second method comprises the following steps: tanswell and Matrigel Transwell experiments
Cell migration and invasion capacity were both measured using Transwell chambers with a pore size of 8 μm. In contrast to the migration experiment, the invasion experiment was performed by spreading a layer of matrigel (0.5mg/mL) on a Tanswell porous filter. 100 μ L HONE1 cells (4X 10)5mL) in RPMI 1640 containing different concentrations of Compound 13(0, 0.03, 0.1, 0.3, 1, 3. mu.M)Medium (serum-free) Transwell upper chamber, lower chamber was 600. mu.L complete medium (10% FBS). After 24h incubation, cells were first fixed with 4% paraformaldehyde and then stained with 0.1% crystal violet for 10 min. The stained cells were then gently wiped off with a cotton swab, air dried at room temperature, and then observed and photographed under a phase contrast microscope.
2. Results of the experiment
As shown in fig. 5, fig. 5A shows that compound 13 can significantly inhibit the healing of HONE1 cell scratch and is concentration-dependent, and that compound 13 has completely lost the ability of HONE1 to heal scratch at 10 μ M, and that the 48-hour effect is more significant than the 24-hour effect, indicating that compound 13 can inhibit the migration of HONE1 cells. From fig. 5B, we can see that: whether in the migration experiment or the invasion experiment, the compound 13 treated group showed a significant decrease in cells passing through the Tanswell membrane and was in a concentration-dependent relationship. The experimental result shows that the compound 13 can inhibit the migration and invasion of cells, and the previous experiment proves that the compound 13 can induce apoptosis, which indicates that the compound 13 is a potential active anticancer compound which can kill tumor cells and inhibit tumor metastasis.
Example 8
The compound (compound 13) of formula (I) was prepared according to the method of example 1, and injection was prepared by adding water for injection, fine-filtering, filling and sterilizing as usual.
Example 9
The compound of formula (I) (compound 13) was prepared as described in example 1, dissolved in sterile water for injection, stirred to dissolve, and filtered through a sterile suction funnel. Then sterile fine filtering, subpackaging in 2 ampoules, freeze drying at low temperature, and aseptically sealing by melting to obtain powder for injection.
Example 10
Compound (compound 13) of formula (I) was prepared according to the method of example 1, and the excipient was added at a ratio of 5:1 by weight to the excipient, followed by granulation and tableting.
Example 11
Compound (compound 13) of formula (I) was prepared according to the method of example 1, and the compound was added to the excipient at a weight ratio of 5:1 to make a capsule.
Example 12
Compound (compound 13) of formula (I) was prepared according to the method of example 1, and the excipient was added in a weight ratio of 3:1 to the excipient to prepare a capsule.
Example 13
The compound (compound 12) represented by the formula (II) was prepared according to the method of example 1, and injection was prepared by adding water for injection, fine-filtering, filling and sterilizing as usual.
Example 14
The compound of formula (II) (Compound 12) was prepared as described in example 1, dissolved in sterile water for injection, stirred to dissolve, and filtered through a sterile suction funnel. Then sterile fine filtering, subpackaging in 2 ampoules, freeze drying at low temperature, and aseptically sealing by melting to obtain powder for injection.
Example 15
Compound (compound 12) of formula (II) was prepared according to the method of example 1, and the excipient was added at a ratio of 5:1 by weight to the excipient, followed by granulation and tableting.
Example 16
Compound (compound 12) of formula (II) was prepared according to the method of example 1, and the compound was added to the excipient at a weight ratio of 5:1 to prepare a capsule.
Example 17
Compound (compound 12) of formula (II) was prepared according to the method of example 1, and the compound was added to the excipient at a weight ratio of 3:1 to prepare a capsule.
Claims (6)
1. A compound extracted from saururus chinensis which is characterized in that the compound is shown as a formula (I),
2. a compound extracted from saururus chinensis which is characterized in that the compound is shown as a formula (II),
3. use of a compound according to claim 1 and/or 2 for the preparation of a medicament for the treatment and/or prophylaxis of nasopharyngeal carcinoma.
4. The use of claim 3, wherein the treatment and/or prevention of nasopharyngeal carcinoma is one or more of inhibiting proliferation of nasopharyngeal carcinoma cells, promoting apoptosis of nasopharyngeal carcinoma cells, or inhibiting migratory invasion of nasopharyngeal carcinoma cells.
5. A medicament for treating and/or preventing nasopharyngeal carcinoma, comprising the compound according to claim 1 and/or 2.
6. The medicament of claim 1, further comprising a pharmaceutically acceptable carrier or excipient.
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