本發(fā)明屬于醫(yī)藥技術(shù)領(lǐng)域，具體涉及米非司酮衍生物及其用途。

背景技術(shù)：

研究發(fā)現(xiàn)，轉(zhuǎn)錄因子KLF5(Kruppel-like factor 5,鋅指蛋白轉(zhuǎn)錄因子5)是與DNA結(jié)構(gòu)域結(jié)合的轉(zhuǎn)錄調(diào)控因子家族。在正常組織中KLF5參與眾多細(xì)胞的生物學(xué)進(jìn)程，包括細(xì)胞的增殖、分化、遷移、炎癥以及潛能性。在癌癥組織中KLF5的表達(dá)和功能均有所改變，并參與調(diào)節(jié)癌細(xì)胞的增殖、調(diào)控、轉(zhuǎn)移、腫瘤微環(huán)境以及腫瘤干細(xì)胞。KLF5主要參與許多控制細(xì)胞增殖的重要信號(hào)通路，無(wú)論在正常細(xì)胞還是在人類許多癌細(xì)胞中都具有促使細(xì)胞增殖的作用。三陰性乳腺癌(Triple Negative Breast Cancer,TNBC)，是雌激素受體(ER)、孕激素受體(PR)和人表皮生長(zhǎng)因子受體(HER2)均為陰性的一種特殊類型的乳腺癌，是乳腺癌中惡性程度最高的一種亞型，約占所有乳腺癌的10％-25％，至今仍未找到有效的靶向治療藥物。在乳腺癌細(xì)胞中，KLF5通過(guò)上調(diào)基因Cyclin A和DNA復(fù)制因子1(CDT1)等促進(jìn)乳腺癌細(xì)胞增殖。也有研究發(fā)現(xiàn)KLF5主要通過(guò)上調(diào)靶基因FGF-BP(Fibroblast growth factor binding protein1)起促進(jìn)細(xì)胞增殖的作用，同時(shí)促進(jìn)ERK激酶活性，穩(wěn)定MKP-1蛋白表達(dá)促進(jìn)細(xì)胞生存。此外，KLF5在乳腺癌的發(fā)生發(fā)展過(guò)程中，起到很重要的作用。乳腺癌的KLF5在ER陰性乳腺癌中普遍高表達(dá)，且KLF5高表達(dá)的患者生存時(shí)間顯著短于KLF5低表達(dá)的病人。前期研究也發(fā)現(xiàn)KLF5轉(zhuǎn)錄因子在Luminal類型的乳腺癌細(xì)胞T47D細(xì)胞中能夠被孕激素所誘導(dǎo)，而該過(guò)程能夠由孕激素受體抑制劑米非司酮(Mifepristone，MIF)所阻斷。

前期新藥篩選結(jié)果發(fā)現(xiàn)，米非司酮(Mifepristone)能夠抑制三陰性乳腺癌細(xì)胞增殖、誘導(dǎo)細(xì)胞凋亡，并且在較高劑量下能夠抑制人源性三陰性乳腺癌移植瘤在免疫缺陷小鼠體內(nèi)的生長(zhǎng)。進(jìn)一步作用機(jī)制研究還發(fā)現(xiàn)米非司酮是通過(guò)誘導(dǎo)三陰性乳腺癌細(xì)胞中微小RNA-153的表達(dá)，從而抑制乳腺癌的增殖、生存和細(xì)胞干性的轉(zhuǎn)錄因子KLF5的表達(dá)(Theranostics 2016,6,533-44)。但是大量研究數(shù)據(jù)和前期活性測(cè)試結(jié)果均顯示米非司酮必須在較高劑量下才能產(chǎn)生抗腫瘤活性，而且米非司酮的主要代謝產(chǎn)物單去甲基米非司酮(RU-42633,美她司酮)則沒(méi)有明顯的抗腫瘤活性。鑒于米非司酮在體內(nèi)很快就代謝為單去甲基米非司酮，而且這個(gè)主要代謝產(chǎn)物在給藥后0.5-2小時(shí)后血液中的濃度遠(yuǎn)超過(guò)母體化合物，大大限制了米非司酮可能作為一種較為理想的靶向抗腫瘤藥物的進(jìn)一步臨床應(yīng)用。

技術(shù)實(shí)現(xiàn)要素：

針對(duì)以上技術(shù)問(wèn)題，因此，本發(fā)明提供一類米非司酮衍生物，以其作為高效抗癌藥物的活性成份，從而為尋找抗三陰性乳腺癌的靶向藥物開(kāi)辟新的途徑。

基、苯基取代苯基、烯烴、苯基取代烯烴或雜環(huán)，包括噻唑、2-噻吩、吡嗪、吡啶、鹵代吡啶；當(dāng)R為磺酰基時(shí)，其具體結(jié)構(gòu)特征為R₂SO₂-，R₂為苯基、C1-C6烷基取代苯基。

優(yōu)選地，當(dāng)R為?；鶗r(shí)，其中R1為氯代C1-C6烷基、鹵代苯基、萘基、硝基取代苯基、C1-C6烷基取代苯基或苯基取代苯基。進(jìn)一步優(yōu)選地，R1為氯代甲烷基。優(yōu)選地，R1為氯或溴代苯基。更加優(yōu)選地，R1為3，4氯代苯基或2-溴4-氯代苯基。

優(yōu)選地，R1為苯基或甲基取代苯基。更加優(yōu)選地，R1為甲基取代3-苯基或甲基取代4-苯基。

本發(fā)明的米非司酮衍生物通過(guò)如下方法制備：

方法a：將1.0當(dāng)量美她司酮和1.5當(dāng)量的酰基氯或磺酰氯加入到0.1-0.2M乙酸乙酯溶液中，在0℃時(shí)加入3.0當(dāng)量的三乙胺，將所得混合物在室溫下攪拌16小時(shí)，然后用乙酸乙酯稀釋該溶液，并用1M鹽酸溶液和鹽水洗滌。用無(wú)水硫酸鈉干燥有機(jī)層，然后在減壓下濃縮，殘余物經(jīng)石油醚/乙酸乙酯比例為4/1:1/1硅膠柱色譜法的柱層析，得到所需產(chǎn)物。

方法b：將1.2-2.0當(dāng)量的一元芳香羧酸，1.0當(dāng)量的美她司酮，1.5-2.0當(dāng)量的苯并三氮唑-N,N,N',N'-四甲基脲六氟磷酸鹽和0.1-0.2M的N,N-二甲基甲酰胺，2.0-4.0當(dāng)量的N，N-二異丙基乙胺混合，然后將該混合物在室溫下攪拌直到美她司酮被反應(yīng)完全，并通過(guò)薄層色譜法測(cè)定；用乙酸乙酯稀釋混合物并用1M鹽酸水溶液淬滅；用水和鹽水洗滌洗滌有機(jī)層后，用無(wú)水硫酸鈉干燥，并減壓濃縮，將粗物質(zhì)通過(guò)硅膠柱色譜法柱層析或用薄層色譜法純化后，得到所需產(chǎn)物。

方法c：將1.2-2.0當(dāng)量的一元芳香羧酸，1.0當(dāng)量的美她司酮和3.0當(dāng)量的1-乙基-(3-二甲基氨基丙基)碳酰二亞胺鹽酸鹽，0.1-0.2M二氯甲烷混合，讓混合物在室溫下攪拌直到美她司酮被反應(yīng)完全，通過(guò)薄層色譜法測(cè)定；用二氯甲烷稀釋混合物并用1M鹽酸水溶液淬滅；用水和鹽水洗滌有機(jī)層，并用無(wú)水硫酸鈉干燥，然后減壓濃縮，將粗物質(zhì)通過(guò)硅膠柱色譜法柱層析或用薄層色譜法純化后，得到所需產(chǎn)物。

其中，所述的美她司酮Metapristone的制備方法參照前期研究結(jié)果及專利CN2013 10510394.0，名稱為“一種合成美她司酮的方法”。

通常用TLC來(lái)跟蹤測(cè)定反應(yīng)的完成程度，反應(yīng)完畢后一般用冰水淬滅，用乙酸乙酯、二氯甲烷、三氯甲烷等萃取，依次用水、飽和食鹽水洗滌，干燥，低溫減壓除去溶劑，經(jīng)柱層析得到最終產(chǎn)物，得到產(chǎn)物用核磁共振、高分辨質(zhì)譜等方法驗(yàn)證其結(jié)構(gòu)。

本發(fā)明的另一目的在于提供本發(fā)明米非司酮衍生物的用途，特別是在制備治療與KLF5調(diào)控相關(guān)腫瘤方面的藥物中的用途。

進(jìn)一步地，本發(fā)明米非司酮衍生物在制備治療三陰性乳腺癌藥物中的用途。

本發(fā)明的有益效果：

1)本發(fā)明在前期探索工作的基礎(chǔ)上，通過(guò)骨架再利用的藥物設(shè)計(jì)原理，以米非司酮代謝敏感結(jié)構(gòu)區(qū)域?yàn)榻Y(jié)構(gòu)改造位點(diǎn)，合理設(shè)計(jì)聚焦型化合物庫(kù)，通過(guò)系統(tǒng)的結(jié)構(gòu)優(yōu)化，設(shè)計(jì)與合成了一類米非司酮衍生物，提供了米非司酮衍生物的結(jié)構(gòu)式，該化合物結(jié)構(gòu)簡(jiǎn)單易于制備且穩(wěn)定，該米非司酮衍生物較米非司酮抗腫瘤活性強(qiáng)。

2)本發(fā)明的米非司酮衍生物作為治療三陰性乳腺癌的藥物活性較米非司酮強(qiáng)，且劑量小。

3)本發(fā)明提供的米非司酮衍生物用于制備成治療三陰性乳腺癌的藥物，為其他同類化合物的合成提供新的解決思路和方案。

具體實(shí)施方式

下述制備實(shí)施例中，用NMR定標(biāo)：δH/C 7.26/77.16ppm(CDCl₃)；試劑主要由安耐吉試劑公司提供，產(chǎn)品純化主要用柱色譜法，硅膠(200-300目或300-400目)，柱色譜法所用硅膠由阿達(dá)瑪斯試劑公司提供(cas：63231-67-4)。

下述縮寫詞：EtOAc:乙酸乙酯；Et₃N：三乙胺；Na₂SO₄：硫酸鈉；PE：石油醚；HBTU：苯并三氮唑-N,N,N',N'-四甲基脲六氟磷酸鹽；DMF：N,N-二甲基甲酰胺；DIPEA：N，N-二異丙基乙胺；TLC：硅膠色譜板；EDCI：1-乙基-(3-二甲基氨基丙基)碳酰二亞胺鹽酸鹽；CH₂Cl₂：二氯甲烷。

實(shí)施例1 FZU-0000-005

2-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylacetamide

將28mg,0.24mmol 2-氯乙酰氯,84mg,0.2mmol美她司酮和42mg,0.4mmol Et₃N加入到2.0mL EtOAc中攪拌16小時(shí)，PE/EtOAc＝1:1，按方法(a)制備得到90mg產(chǎn)物，得率92％，產(chǎn)物為白色固體，mp:124.3–125.5℃。

所得的產(chǎn)物進(jìn)行氫譜¹HNMR、碳譜¹³CNMR及HRMS檢測(cè)，結(jié)果如下：¹H NMR(400MHz,CDCl₃)δ7.27(d,J＝6.0Hz,2H),7.16(d,J＝7.7Hz,2H),5.80(s,1H),4.47(d,J＝6.9Hz,1H),3.82(s,2H),3.27(s,3H),2.85–2.75(m,1H),2.67–2.56(m,2H),2.53–2.38(m,4H),2.37–2.15(m,4H),2.10–2.01(m,1H),2.01–1.94(m,1H),1.91(s,3H),1.79–1.72(m,2H),1.56–1.47(m,1H),1.41–1.32(m,1H),0.50(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.40,166.42,156.52,145.54,144.96,140.31,129.93,128.68,127.12,123.32,82.75,82.21,80.03,49.75,46.96,41.54,40.19,39.27,39.16,38.96,38.08,36.81,31.13,27.36,25.99,23.40,13.97,3.94.

HRMS(ESI-TOF):calcd for C₃₀H₃₅ClNO₃[M+H]⁺492.2300；found 492.2290.

實(shí)施例2

2-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylpropan amide(FZU-0010-001)

與實(shí)施例1不同之處在于：2-氯乙酰氯為30mg,Et₃N為40mg，得到80mg產(chǎn)物，得率為79％，其中PE/EtOAc＝2:1.產(chǎn)物為白色固體，mp:124.6–125.6℃。

¹H NMR(400MHz,CDCl₃)δ7.27(d,J＝7.9Hz,2H),7.18(d,J＝8.0Hz,2H),5.80(s,1H),4.48(d,J＝6.8Hz,1H),4.26(q,J＝6.3Hz,1H),3.28(s,3H),2.90–2.74(m,1H),2.68–2.58(m,2H),2.54–2.38(m,4H),2.32–2.19(m,2H),2.13–1.95(m,4H),1.91(s,3H),1.83–1.67(m,2H),1.56(d,J＝6.5Hz,3H),1.53–1.44(m,1H),1.40–1.28(m,1H),0.51(s,3H).¹³C NMR(101MHz,CDCl₃)δ199.43,169.57,156.55,145.37,145.09,140.51,129.98,128.61,127.30,123.37,82.82,82.26,80.11,49.85,47.01,40.25,39.29,39.19,39.02,38.15,36.85,31.16,27.41,26.04,23.44,21.36,14.00,13.99,3.94.

HRMS(ESI-TOF):calcd for C₃₁H₃₇ClNO₃[M+H]⁺506.2456；found 506.2456.

實(shí)施例3

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N,4-dimethylbenzenesulfo namide(FZU-0025-008)

與實(shí)施例1不同之處在于：用46mg，0.24mmol的對(duì)甲基苯磺酰氯替換28mg,0.24mmol 2-氯乙酰氯，根據(jù)方法a得到83mg白色固體，收率73％，mp:121.8–122.9℃。

¹H NMR(400MHz,CDCl₃)δ7.33(d,J＝7.9Hz,2H),7.19(d,J＝7.9Hz,2H),7.11(d,J＝8.2Hz,2H),7.01(d,J＝8.2Hz,2H),5.78(s,1H),4.42(d,J＝6.8Hz,1H),3.11(s,3H),2.87–2.66(m,1H),2.66–2.56(m,2H),2.51–2.36(m,6H),2.34–2.18(m,4H),2.14–1.97(m,3H),1.91(s,3H),1.82–1.68(m,2H),1.55–1.44(m,1H),1.35–1.28(m,1H),0.51(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.42,156.61,145.44,143.74,143.60,139.14,133.03,129.62,129.24,127.81,127.34,126.24,123.07,82.65,82.20,80.02,49.71,46.93,40.07,39.17,39.12,38.87,37.78,36.78,31.08,27.31,25.84,23.34,21.57,13.74,3.86.

HRMS(ESI-TOF):calcd for C₃₅H₄₀NO₄S[M+H]⁺592.2492；found 592.2472.

實(shí)施例4

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzenesulfonamide(FZU-0025-009)

與實(shí)施例1不同之處在于：用113mg，0.64mmol的苯磺酰氯替換28mg,0.24mmol 2-氯乙酰氯，將作為起始原料，根據(jù)方法a.得到63mg白色固體，收率57％，mp:120.3–121.5℃℃。

¹H NMR(400MHz,CDCl₃)δ7.58–7.54(m,1H),7.46(d,J＝7.2Hz,2H),7.40(d,J＝7.9Hz,2H),7.15–7.07(m,2H),7.02–6.96(m,2H),5.79(s,1H),4.42(d,J＝6.8Hz,1H),3.15(s,3H),2.85–2.73(m,1H),2.65–2.57(m,2H),2.54–2.35(m,5H),2.32–2.20(m,3H),2.11–1.98(m,3H),1.88(s,3H),1.80–1.67(m,2H),1.57–1.43(m,1H),1.40–1.31(m,1H),0.51(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.40,156.58,145.35,143.87,138.98,135.93,132.82,129.65,128.99,128.92,128.62,127.74,127.39,127.32,126.25,124.95,123.10,82.68,82.19,80.02,49.71,46.93,40.05,39.18,39.10,38.88,37.86,36.78,31.07,27.31,25.84,23.34,13.77,3.86.

HRMS(ESI-TOF):calcd for C₃₄H₃₈NO₄S[M+H]⁺578.2336；found 578.2314.

實(shí)施例5

3,4-Dichloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0000-008)

取8mg,0.24mmol 3,4-二氯苯甲酸、84mg,0.2mmol的美她司酮、112mg,0.6mmol的HBTU和76mg,0.6mmol DIPEA加入2.0mL DMF中攪拌16小時(shí)，PE/EtOAc＝2:1，按方法b制備得到84mg產(chǎn)物，得率為71％，產(chǎn)物為白色固體，mp:230.1–231.5℃。

¹H NMR(400MHz,CDCl₃)δ7.32(s,1H),7.12(d,J＝8.3Hz,1H),7.04(d,J＝8.0Hz,2H),7.01(d,J＝8.6Hz,1H),6.89(d,J＝7.9Hz,2H),5.72(s,1H),4.33(d,J＝6.9Hz,1H),3.42(s,3H),2.75–2.62(m,1H),2.60–2.48(m,2H),2.46–2.24(m,5H),2.23–2.09(m,3H),2.02–1.88(m,2H),1.83(s,3H),1.76–1.60(m,2H),1.49–1.36(m,1H),1.35–1.24(m,1H),0.38(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.30,167.82,156.47,145.04,144.11,141.82,135.51,133.91,131.97,130.98,129.70,129.59,128.17,128.10,126.91,123.15,82.47,82.25,79.92,49.57,46.87,40.01,39.21,39.06,38.85,38.33,36.70,31.08,27.32,25.77,23.31,13.73,3.87.

HRMS(ESI-TOF):calcd for C₃₅H₃₆Cl₂NO₃[M+H]⁺588.2067；found 588.2058.

實(shí)施例6

3-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenza mide(FZU-0002-016)

與實(shí)施例5不同之處在于：將8mg,0.24mmol 3,4-二氯苯甲酸替換為47mg,0.3mmol3-氯苯甲酸，HBTU為152mg,0.4mmol，DIPEA為103mg,0.8mmol，得100mg產(chǎn)物，得率為90％。產(chǎn)物為白色固體，mp:122.6–123.7℃。

¹H NMR(400MHz,CDCl₃)δ7.27(s,1H),7.19(d,J＝7.7Hz,1H),7.15–6.99(m,4H),6.94(d,J＝7.7Hz,2H),5.77(s,1H),4.36(d,J＝7.2Hz,1H),3.45(s,3H),2.80–2.67(m,1H),2.65–2.53(m,2H),2.51–2.30(m,4H),2.28–2.13(m,3H),2.05–1.97(m,1H),1.96–1.91(m,1H),1.89(s,3H),1.84–1.80(m,1H),1.80–1.66(m,2H),1.54–1.42(m,1H),1.38–1.27(m,1H),0.36(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.43,168.98,156.54,145.17,143.90,142.12,137.54,133.74,129.80,129.05,128.95,128.02,127.06,127.02,123.23,82.72,82.25,80.09,49.65,46.91,40.10,39.30,39.19,38.92,38.70,38.29,36.81,31.17,27.41,25.83,23.39,13.78,3.93.

HRMS(ESI-TOF):calcd for C₃₅H₃₅ClNO₃[M-H]^-552.2311；found 552.2293.

實(shí)施例7

3,6-Dichloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylpicolinamide(FZU-0010-006)

與實(shí)施例5不同之處在于：將8mg,0.24mmol 3,4-二氯苯甲酸替換為46mg,0.24mmol 3,6-二氯吡啶甲酸，HBTU為114mg,0.3mmol，DIPEA為52mg,0.4mmol，PE/EtOAc＝1:1，得164mg產(chǎn)物，得率為99％。產(chǎn)物為白色固體，mp:126.5–127.6℃。

¹H NMR(400MHz,CDCl₃)δ7.41(d,J＝8.4Hz,1H),7.15–7.03(m,3H),7.03–6.96(m,2H),5.75(s,1H),4.30(d,J＝7.2Hz,1H),3.46(s,3H),2.72–2.60(m,1H),2.60–2.51(m,2H),2.48–2.26(m,4H),2.24–2.07(m,3H),2.05–1.95(m,2H),1.95–1.90(m,1H),1.87(s,3H),1.77–1.63(m,2H),1.49–1.37(m,1H),1.35–1.25(m,1H),0.21(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.40,164.94,156.52,153.28,148.52,145.11,144.81,139.73,139.53,129.79,127.68,127.38,127.10,124.98,123.23,82.77,82.18,80.05,49.61,46.78,40.12,39.39,39.17,38.86,36.80,36.68,31.13,27.39,25.81,23.39,13.32,3.90.

HRMS(ESI-TOF):calcd for C₃₄H₃₅Cl₂N₂O₃[M+H]⁺589.2019；found 589.2020.

實(shí)施例8

5-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylnicotinamide(FZU-0010-008)

與實(shí)施例7不同之處在于：將46mg,0.24mmol的3,6-二氯吡啶甲酸替換為38mg,0.24mmol的5-氯煙酸，得到109mg產(chǎn)物，得率為98％，產(chǎn)物為白色固體，mp:118.9–120.1℃℃。

¹H NMR(400MHz,CDCl₃)δ8.37(s,1H),8.32(s,1H),7.50(s,1H),7.08(d,J＝8.0Hz,2H),6.94(d,J＝7.9Hz,2H),5.75(s,1H),4.34(d,J＝7.1Hz,1H),3.46(s,3H),2.76–2.64(m,1H),2.60–2.50(m,2H),2.49–2.31(m,4H),2.29–2.26(m,1H),2.24–2.10(m,4H),2.08–1.93(m,3H),1.93–1.88(m,1H),1.86(s,3H),1.67(dt,J＝11.4,9.4Hz,2H),1.48–1.38(m,1H),1.35–1.27(m,1H),0.38(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.39,166.53,156.47,149.24,147.41,144.93,144.66,141.38,135.87,132.77,131.26,129.87,128.39,127.23,123.30,82.64,82.26,80.04,49.66,46.90,40.10,39.26,39.15,38.95,38.32,36.79,31.15,27.38,25.86,23.38,13.86,3.92.

HRMS(ESI-TOF):calcd for C₃₄H₃₆ClN₂O₃[M+H]⁺555.2409；found 555.2397.

實(shí)施例9

5-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylpicolinamide(FZU-0010-009)

與實(shí)施例8不同之處在于：5-氯煙酸為45mg,0.28mmol，HBTU為152mg,0.4mmol，得到120mg產(chǎn)物，得率為90％。產(chǎn)物為白色固體，mp:124.1–125.5℃。

¹H NMR(400MHz,CDCl₃)δ8.17(s,1H),7.54(d,J＝6.6Hz,1H),7.48–7.35(m,1H),7.14–6.84(m,4H),5.77(s,1H),4.36(d,J＝6.8Hz,1H),3.48(s,3H),2.79–2.66(m,1H),2.62–2.53(m,2H),2.51–2.29(m,4H),2.28–2.16(m,3H),2.14–2.07(m,1H),2.06–1.90(m,3H),1.88(s,3H),1.80–1.66(m,1H),1.53–1.41(m,1H),1.38–1.28(m,1H),0.35(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.40,167.71,156.55,152.17,147.14,145.25,143.64,141.87,135.98,132.38,129.72,127.74,126.90,124.76,123.20,82.69,82.26,80.04,49.70,46.95,40.07,39.30,39.13,38.93,37.90,36.81,31.13,27.37,25.86,23.39,13.63,3.92.

實(shí)施例10

4-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylpicolinamide(FZU-0010-010)

與實(shí)施例9不同之處在于：將5-氯煙酸替換為4-氯煙酸，得到140mg產(chǎn)物，產(chǎn)率為90％，PE/EtOAc＝1:2。產(chǎn)物為白色固體，mp:121.3–122.5℃。

¹H NMR(400MHz,CDCl₃)δ8.12(d,J＝8.0Hz,1H),7.46(s,1H),7.12(d,J＝8.0Hz,1H),7.03(d,J＝8.0Hz,2H),6.95(d,J＝8.0Hz,2H),5.77(s,1H),4.36(d,J＝6.3Hz,1H),3.50(s,3H),2.80–2.65(m,1H),2.65–2.52(m,2H),2.51–2.30(m,4H),2.28–2.10(m,4H),2.06–1.93(m,2H),1.88(s,3H),1.80–1.57(m,2H),1.53–1.42(m,1H),1.35–1.23(m,1H),0.36(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.44,167.41,156.59,155.52,149.18,145.29,144.40,143.74,141.75,129.72,127.79,126.86,124.36,124.28,123.19,82.72,82.25,80.05,49.67,46.90,40.09,39.27,39.17,38.89,37.93,36.82,31.14,27.38,25.83,23.38,13.63,3.91.

HRMS(ESI-TOF):calcd for C₃₄H₃₆ClN₂O₃[M+H]⁺555.2409；found 588.2401.

實(shí)施例11

6-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylnicotinamide(FZU-0010-012)

與實(shí)施例9不同之處在于：將5-氯煙酸替換為6-氯煙酸，得到116mg產(chǎn)物，產(chǎn)率為90％，

PE/EtOAc＝2：3。產(chǎn)物為白色固體，mp:127.2–128.4℃。

¹H NMR(400MHz,CDCl₃)δ8.27(s,1H),7.41(d,J＝8.0Hz,1H),7.07(d,J＝8.1Hz,2H),7.03(d,J＝8.3Hz,1H),6.93(d,J＝8.0Hz,2H),5.75(s,1H),4.35(d,J＝7.0Hz,1H),3.45(s,3H),2.71(dt,J＝14.8,5.0Hz,1H),2.60–2.48(m,2H),2.48–2.28(m,4H),2.27–2.12(m,4H),2.05–1.92(m,3H),1.86(s,3H),1.76–1.62(m,2H),1.51–1.38(m,1H),1.35–1.24(m,1H),0.36(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.40,167.01,156.48,152.17,150.02,144.90,144.55,141.54,138.75,130.63,129.88,128.35,127.28,123.30,123.20,82.69,82.20,80.03,49.69,46.97,40.04,39.29,39.06,38.96,38.28,36.77,31.11,27.34,25.88,23.36,13.85,3.93.

HRMS(ESI-TOF):calcd for C₃₄H₃₆ClN₂O₃[M+H]⁺555.2409；found 555.2401.

實(shí)施例12

2-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylisonicotinamide(FZU-0010-014)

與實(shí)施例9不同之處在于：將5-氯煙酸替換為2-氯煙酸，得到100mg產(chǎn)物，產(chǎn)率為90％，PE/EtOAc＝2：3。產(chǎn)物為白色固體，mp:117.5–118.6℃。

¹H NMR(400MHz,CDCl₃)δ8.30(s,1H),7.44(d,J＝7.9Hz,1H),7.11(d,J＝6.9Hz,2H),7.06(d,J＝7.4Hz,1H),6.96(d,J＝6.8Hz,2H),5.78(s,1H),4.38(d,J＝5.4Hz,1H),3.49(s,3H),2.81–2.68(m,1H),2.64–2.54(m,2H),2.52–2.30(m,4H),2.29–2.17(m,4H),2.09–1.94(m,3H),1.90(s,3H),1.79–1.65(m,2H),1.53–1.41(m,1H),1.40–1.29(m,1H),0.36(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.38,167.01,156.46,152.17,150.04,144.89,144.54,141.55,138.74,130.63,129.88,128.35,127.29,123.31,123.19,82.70,82.20,80.03,49.70,46.97,40.04,39.28,39.06,38.96,38.28,36.78,31.11,27.34,25.88,23.36,13.84,3.93.

HRMS(ESI-TOF):calcd for C₃₄H₃₆ClN₂O₃[M+H]⁺555.2409；found 555.2401.

實(shí)施例13

Following the General Procedure C:

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylpicolinamide(FZU-0002-018)

將37mg,0.3mmol的吡啶甲酸，84mg,0.2mmol的美她司酮，115mg,0.6mmol的EDCI加入到2.0mL CH₂Cl₂中，按方法C制備得到102mg的產(chǎn)物，得率為98％，PE/EtOAc＝1:3，產(chǎn)物為黃色固體，mp:116.3–117.4℃。

¹H NMR(400MHz,CDCl₃)δ8.36–8.15(m,1H),7.60–7.44(m,1H),7.41–7.28(m,1H),7.17–6.74(m,5H),5.73(s,1H),4.31(d,J＝6.6Hz,1H),3.47(s,3H),2.74–2.62(m,1H),2.60–2.49(m,2H),2.46–2.25(m,4H),2.23–2.08(m,5H),2.06–1.91(m,2H),1.85(s,3H),1.76–1.59(m,2H),1.51–1.35(m,1H),1.33–1.21(m,1H),0.34(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.49,168.79,156.64,154.21,148.37,145.42,143.41,141.99,136.19,129.65,127.65,126.87,123.93,123.75,123.12,82.69,82.25,80.03,49.65,46.89,40.07,39.27,39.16,38.87,37.79,36.81,31.12,27.35,25.80,23.38,13.66,3.90.

HRMS(ESI-TOF):calcd for C₃₄H₃₅N₂O₃[M-H]^-519.2653；found 519.2623.

實(shí)施例14

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0002-006)

將44mg,0.36mmol的吡啶甲酸，125mg,0.3mmol的美她司酮，173mg,0.9mmol的EDCI加入到3.0mL CH₂Cl₂中，按方法C制備得到120mg的產(chǎn)物，得率為77％，PE/EtOAc＝1:1，產(chǎn)物為白色固體，mp:126.5–127.9℃。

¹H NMR(400MHz,CDCl₃)δ7.24(d,J＝7.8Hz,2H),7.20(d,J＝7.1Hz,1H),7.10(t,J＝7.4Hz,2H),7.03(d,J＝8.0Hz,2H),6.93(d,J＝7.9Hz,2H),5.76(s,1H),4.35(d,J＝7.1Hz,1H),3.49(s,3H),2.71(dt,J＝10.8,5.1Hz,1H),2.63–2.52(m,2H),2.51–2.30(m,4H),2.29–2.16(m,4H),2.07–1.93(m,3H),1.90(s,3H),1.79–1.66(m,2H),1.51–1.41(m,1H),1.37–1.28(m,1H),0.40(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.44,170.57,156.56,145.35,143.47,142.60,135.78,129.69,129.67,128.87,127.81,127.61,127.09,123.16,82.66,82.28,80.06,49.67,46.93,40.09,39.34,39.17,38.90,38.22,36.80,31.13,27.38,25.80,23.39,13.75,3.90.

HRMS(ESI-TOF):calcd for C₃₅H₃₆NO₃[M-H]^-518.2701；found 519.2693.

實(shí)施例15

3-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylpropanamide(FZU-0002-014)

與實(shí)施例14不同之處在于：將44mg,0.36mmol的吡啶甲酸替換為39mg,0.36mmol 3-氯丙酸，得到147mg產(chǎn)物，產(chǎn)率為97％，PE/EtOAc＝1:1，產(chǎn)物為白色固體。

¹H NMR(400MHz,CDCl₃)δ7.26(d,J＝8.2Hz,2H),7.11(d,J＝8.4Hz,2H),5.80(s,1H),4.47(d,J＝7.0Hz,1H),3.80–3.67(m,2H),3.27(s,3H),2.81(dt,J＝10.1,5.6Hz,1H),2.65–2.57(m,2H),2.54–2.41(m,5H),2.37–2.22(m,3H),2.09–2.02(m,1H),2.01–1.92(m,1H),1.90(s,3H),1.79–1.71(m,2H),1.52–1.46(m,1H),1.40–1.31(m,1H),0.51(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.33,169.64,156.47,145.11,145.00,141.08,129.91,128.58,127.44,123.32,82.71,82.30,80.06,49.82,47.00,40.21,40.16,39.28,39.18,39.00,37.40,36.96,36.85,31.14,27.39,26.01,23.42,13.95,3.92.

HRMS(ESI-TOF):calcd for C₃₁H₃₅ClNO₃[M-H]^-504.2311；found 504.2322.

實(shí)施例16

4-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0002-015)

與實(shí)施例14不同之處在于：將吡啶甲酸替換為4-氯苯甲酸，得到140mg產(chǎn)物，產(chǎn)率為90％，產(chǎn)物為白色固體，mp:117.4–118.6℃。

¹H NMR(400MHz,CDCl₃)δ7.18(d,J＝8.5Hz,2H),7.12–6.99(m,4H),6.92(d,J＝8.4Hz,2H),5.77(s,1H),4.37(d,J＝7.1Hz,1H),3.46(s,3H),2.73(dt,J＝15.0,5.3Hz,1H),2.63–2.49(m,2H),2.49–2.29(m,4H),2.28–2.17(m,3H),2.09–1.92(m,3H),1.89(s,3H),1.80–1.66(m,2H),1.53–1.39(m,1H),1.37–1.26(m,1H),0.37(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.36,169.41,156.48,145.13,143.82,142.36,135.76,134.23,130.39,129.80,128.01,127.92,127.09,123.27,82.74,82.26,80.09,49.72,46.99,40.10,39.39,39.15,38.98,38.32,36.82,31.15,27.41,25.88,23.41,13.76,3.93.

HRMS(ESI-TOF):calcd for C₃₅H₃₅ClNO₃[M-H]^-552.2311；found 552.2292.

實(shí)施例17

3-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N,4-dimethylbenzamide(FZU-0010-021)

與實(shí)施例13不同之處在于：將37mg,0.3mmol的吡啶甲酸替換為51mg,0.3mmol4-甲基-3-氯苯甲酸，PE/EtOAc＝3:2，得到產(chǎn)物為70mg,得率為61％。產(chǎn)物為白色固體，mp:116.6–117.8℃。

¹H NMR(400MHz,CDCl₃)δ7.24(s,1H),7.07(d,J＝8.3Hz,2H),7.03(dd,J＝7.9,1.6Hz,1H),7.01–6.87(m,4H),5.77(s,1H),4.37(d,J＝7.3Hz,1H),3.47(s,3H),2.77–2.66(m,1H),2.63–2.53(m,2H),2.49–2.36(m,3H),2.36–2.31(m,1H),2.30–2.25(m,3H),2.23–2.14(m,3H),2.07–1.96(m,2H),1.91(s,3H),1.80–1.68(m,2H),1.53–1.43(m,1H),1.38–1.29(m,1H),0.44(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.40,169.01,156.53,145.29,143.84,142.39,137.95,134.83,133.71,130.12,129.77,129.67,128.03,127.25,127.02,123.21,82.69,82.30,80.09,49.64,46.94,40.18,39.37,39.28,38.94,38.38,36.81,31.18,27.44,25.85,23.42,20.03,13.75,3.90.

實(shí)施例18

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-3-methoxy-N-methylbenzamide(FZU-0010-022)

與實(shí)施例17不同之處在于：將51mg,0.3mmol4-甲基-3-氯苯甲酸替換為61mg,0.3mmol的3-甲氧基苯甲酸，得到76mg的產(chǎn)物，得率為69％，產(chǎn)物為白色固體，mp:112.6–113.8℃。

¹H NMR(400MHz,CDCl₃)δ7.09–7.01(m,2H),7.00–6.91(m,3H),6.88(s,1H),6.75(d,J＝7.7Hz,2H),5.76(s,1H),4.35(d,J＝6.5Hz,1H),3.67(s,3H),3.48(s,3H),2.78–2.64(m,1H),2.62–2.51(m,2H),2.50–2.30(m,4H),2.28–2.16(m,3H),2.11–1.94(m,3H),1.86(s,3H),1.81–1.65(m,2H),1.53–1.40(m,1H),1.37–1.28(m,1H),0.41(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.42,170.40,158.97,156.56,145.38,143.52,142.61,137.09,129.71,128.60,127.83,126.96,123.17,121.27,115.66,114.12,82.67,82.30,80.08,55.22,49.66,46.91,40.15,39.36,39.25,38.92,38.24,36.81,31.16,27.41,25.82,23.41,13.68,3.89.

HRMS(ESI-TOF):calcd for C₃₆H₄₀NO₄[M+H]⁺550.2952；found 550.2945.

實(shí)施例19

4-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methyl-2-nitrobenzamide(FZU-0010-023)

與實(shí)施例17不同之處在于：將51mg,0.3mmol的4-甲基-3-氯苯甲酸替換為80mg,0.4mmol的4-氯-2-硝基苯甲酸，PE/EtOAc＝1:1得到81mg的產(chǎn)物，得率為68％，產(chǎn)物為白色固體，mp:128.5–129.6℃。

¹H NMR(400MHz,CDCl₃)δ7.81(d,J＝1.5Hz,1H),7.40(dd,J＝8.2,1.5Hz,1H),7.22(d,J＝8.2Hz,1H),6.95(q,J＝8.4Hz,4H),5.73(s,1H),4.29(d,J＝7.0Hz,1H),3.47(s,3H),2.72–2.61(m,1H),2.57–2.48(m,2H),2.45–2.23(m,6H),2.21–2.05(m,3H),2.05–1.95(m,1H),1.95–1.87(m,1H),1.82(s,3H),1.76–1.60(m,2H),1.49–1.37(m,1H),1.33–1.24(m,1H),0.20(s,3H).¹³C NMR(101MHz,CDCl₃)δ199.22,166.14,156.40,146.22,144.89,144.86,140.11,135.18,133.36,131.65,130.55,129.70,128.00,127.02,124.20,123.13,82.43,82.19,76.84,49.53,46.76,39.97,39.24,38.98,38.81,37.00,36.66,31.00,27.26,25.70,23.28,13.35,3.79.

HRMS(ESI-TOF):calcd for C₃₅H₃₆ClN₂O₅[M+H]⁺599.2307；found 599.2294.

實(shí)施例20

3,5-Dichloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-024)

與實(shí)施例19不同之處在于：將80mg,0.4mmol的4-氯-2-硝基苯甲酸替換為76mg,0.4mmol3,5-二氯苯甲酸，得到73mg的產(chǎn)物，得率為61％。產(chǎn)物為白色固體，mp:115.7–116.9℃。

¹H NMR(400MHz,CDCl₃)δ7.19(t,J＝1.8Hz,1H),7.14–7.09(m,2H),7.08(d,J＝8.3Hz,2H),6.92(d,J＝8.4Hz,2H),5.75(s,1H),4.35(d,J＝7.2Hz,1H),3.44(s,3H),2.75–2.62(m,1H),2.59–2.48(m,2H),2.48–2.26(m,4H),2.24–2.07(m,3H),2.07–1.95(m,2H),1.88(s,3H),1.77–1.62(m,2H),1.52–1.39(m,1H),1.37–1.26(m,1H),0.44(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.33,167.50,156.45,145.03,144.32,141.71,138.64,134.49,129.86,129.72,128.24,127.40,126.99,123.30,82.71,82.29,80.11,49.66,46.90,40.16,39.23,38.95,38.39,36.82,31.20,27.45,25.86,23.40,13.79,3.92.

HRMS(ESI-TOF):calcd for C₃₅H₃₆Cl₂NO₃[M+H]⁺588.2067；found 588.2058.

實(shí)施例21

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-2-methoxy-N-methylbenzamide(FZU-0010-026)

與實(shí)施例20不同之處在于：將76mg,0.4mmol3,5-二氯苯甲酸替換為61mg,0.3mmol的甲氧基苯甲酸，PE/EtOAc＝2:3，得到75mg的產(chǎn)物，得率為68％。產(chǎn)物為白色固體，mp:124.2–125.3℃。

¹H NMR(400MHz,CDCl₃)δ7.19–7.05(m,2H),7.03–6.87(m,4H),6.74(t,J＝7.1Hz,1H),6.57(d,J＝8.3Hz,1H),5.75(s,1H),4.29(d,J＝6.7Hz,1H),3.61(s,3H),3.46(s,3H),2.77–2.62(m,1H),2.60–2.49(m,2H),2.49–2.27(m,4H),2.27–2.08(m,4H),2.03–1.91(m,2H),1.87(s,3H),1.79–1.61(m,2H),1.51–1.39(m,1H),1.36–1.27(m,1H),0.29(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.42,169.31,156.55,155.11,145.48,143.39,141.69,130.28,129.64,128.72,127.00,126.85,126.59,123.11,120.16,110.48,82.66,82.27,80.06,55.16,49.63,46.81,40.12,39.28,39.21,38.90,36.92,36.82,31.13,27.39,25.80,23.39,13.56,3.87.

HRMS(ESI-TOF):calcd for C₃₆H₄₀NO₄[M+H]⁺550.2952；found 550.2942.

實(shí)施例22

4-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methyl-3-nitrobenzamide(FZU-0010-027)

與實(shí)施例20不同之處在于：將76mg,0.4mmol3,5-二氯苯甲酸替換為81mg,0.4mmol的4-氯-3-硝基苯甲酸，得到90mg的產(chǎn)物，得率為74％。產(chǎn)物為白色固體，mp:123.3–124.6℃℃℃℃。

¹H NMR(400MHz,CDCl₃)δ7.78(s,1H),7.41(d,J＝8.3Hz,1H),7.30(d,J＝8.7Hz,1H),7.13(d,J＝7.9Hz,2H),6.97(d,J＝7.8Hz,2H),5.78(s,1H),4.38(d,J＝7.0Hz,1H),3.49(s,3H),2.81–2.68(m,1H),2.64–2.53(m,2H),2.51–2.30(m,4H),2.29–2.14(m,3H),2.09–1.98(m,2H),1.98–1.91(m,1H),1.87(s,3H),1.79–1.65(m,2H),1.55–1.41(m,1H),1.38–1.25(m,1H),0.38(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.32,166.68,156.39,147.10,144.73,144.66,141.52,135.61,133.33,131.28,129.95,128.45,128.38,127.08,126.33,123.36,82.74,82.19,80.04,49.68,46.95,40.05,39.25,39.08,38.98,38.55,36.79,31.14,27.39,25.87,23.37,13.69,3.94.

HRMS(ESI-TOF):calcd for C₃₅H₃₅ClN₂NaO₅[M+Na]⁺621.2127；found 621.2166.

實(shí)施例23

4-Chloro-3-fluoro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-028)

與實(shí)施例22不同之處在于：81mg,0.4mmol的4-氯-3-硝基苯甲酸為52mg,0.3mmol的3-氯-4-氟苯甲酸，得到75mg的產(chǎn)物，得率為64％。產(chǎn)物為白色固體，mp:121.2–122.2℃。

¹H NMR(400MHz,CDCl₃)δ7.13(d,J＝7.8Hz,1H),7.11–7.06(m,2H),7.04(dd,J＝9.5,1.5Hz,1H),6.95(dd,J＝11.9,9.3Hz,3H),5.77(s,1H),4.37(d,J＝7.1Hz,1H),3.45(d,J＝8.8Hz,3H),2.73(dt,J＝14.9,5.2Hz,1H),2.65–2.53(m,2H),2.52–2.29(m,4H),2.28–2.17(m,3H),2.17–2.08(m,1H),2.08–1.93(m,2H),1.89(s,3H),1.80–1.64(m,2H),1.53–1.41(m,1H),1.38–1.29(m,1H),0.37(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.27,168.00,158.48,156.41,155.99,145.02,144.21,141.96,136.17,136.11,129.89,129.81,128.16,127.00,125.47,125.43,123.26,122.71,122.53,117.44,117.21,82.63,82.27,80.04,49.68,46.94,40.10,39.35,39.13,38.97,38.34,36.78,31.13,27.39,25.85,23.38,13.62,3.90.

HRMS(ESI-TOF):calcd for C₃₅H₃₆ClFNO₃[M+H]⁺572.2362；found 572.2355.

實(shí)施例24

3-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methyl-2-nitrobenzamide(FZU-0010-029)

與實(shí)施例22不同之處在于：4-氯-3-硝基苯甲酸為60mg,0.3mmol，得到74mg的產(chǎn)物，得率為61％。產(chǎn)物為白色固體，mp:121.8–123.0℃。

¹H NMR(400MHz,CDCl₃)δ7.34(d,J＝8.0Hz,1H),7.15(d,J＝7.9Hz,2H),7.11–6.95(m,3H),6.87(d,J＝7.6Hz,1H),5.77(s,1H),4.36(d,J＝7.0Hz,1H),3.46(s,3H),2.82–2.66(m,1H),2.65–2.53(m,2H),2.50–2.28(m,4H),2.26–2.11(m,3H),2.09–1.99(m,2H),1.97–1.91(m,1H),1.89(s,3H),1.80–1.63(m,2H),1.53–1.41(m,1H),1.35–1.20(m,1H),0.31(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.38,165.01,156.50,147.52,144.98,144.79,140.82,133.13,131.42,130.31,129.85,128.27,127.52,126.75,126.48,123.28,82.84,82.17,80.05,49.65,46.93,40.12,39.42,39.17,38.89,37.57,36.81,31.15,27.40,25.85,23.38,13.68,3.92.

實(shí)施例25

3-Chloro-4-fluoro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-030)

與實(shí)施例23不同之處在于：3-氯-4-氟苯甲酸為70mg,0.4mmol，得到80mg的產(chǎn)物，得率為68％。產(chǎn)物為白色固體，mp:118.6–119.6℃。

¹H NMR(400MHz,CDCl₃)δ7.35(dd,J＝7.0,2.1Hz,1H),7.16–7.11(m,1H),7.09(d,J＝8.3Hz,2H),6.94(d,J＝8.4Hz,2H),6.86(t,J＝8.6Hz,1H),5.77(s,1H),4.38(d,J＝7.1Hz,1H),3.46(s,3H),2.73(dt,J＝14.8,5.2Hz,1H),2.64–2.53(m,2H),2.51–2.30(m,4H),2.28–2.16(m,4H),2.08–1.98(m,2H),1.91(s,3H),1.81–1.67(m,2H),1.51–1.41(m,1H),1.39–1.30(m,1H),0.43(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.38,167.98,159.84,157.33,156.53,145.10,144.04,142.12,132.82,132.78,131.74,129.79,129.26,129.19,128.13,126.98,123.23,120.66,120.48,115.92,115.71,82.64,82.28,80.04,49.69,46.95,40.09,39.29,39.14,38.95,38.47,36.77,31.14,27.38,25.84,23.37,13.80,3.89.

HRMS(ESI-TOF):calcd for C₃₅H₃₆ClFNO₃[M+H]⁺572.2362；found 572.2355.

實(shí)施例26

4-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-2-methoxy-N-methylbenzamide(FZU-0010-032)

與實(shí)施例25不同之處在于：將70mg,0.4mmol的3-氯-4-氟苯甲酸替換為75mg,0.4mmol的2-氯-4-甲氧基苯甲酸，得到82mg的產(chǎn)物，得率為69％。產(chǎn)物為白色固體,mp:122.6–123.8℃。

¹H NMR(400MHz,CDCl₃)δ7.05(d,J＝7.8Hz,1H),7.00–6.83(m,4H),6.74(d,J＝7.7Hz,1H),6.57(s,1H),5.77(s,1H),4.32(d,J＝6.4Hz,1H),3.62(s,3H),3.45(s,3H),2.76–2.63(m,1H),2.62–2.51(m,2H),2.50–2.28(m,4H),2.27–2.12(m,3H),2.09–1.96(m,2H),1.96–1.92(m,1H),1.89(s,3H),1.79–1.65(m,2H),1.53–1.40(m,1H),1.38–1.27(m,1H),0.29(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.37,168.26,156.53,155.82,145.26,143.71,141.40,135.71,129.71,129.60,127.17,126.84,125.27,123.18,120.34,111.31,82.65,82.22,80.06,55.49,49.67,46.86,40.09,39.32,39.15,38.98,36.96,36.81,31.13,27.39,25.85,23.39,13.56,3.89.

HRMS(ESI-TOF):calcd for C₃₆H₃₉ClNO₄[M+H]⁺584.2562；found 584.2544.

實(shí)施例27

3-Bromo-4-chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-036)

與實(shí)施例25不同之處在于：將70mg,0.4mmol的3-氯-4-氟苯甲酸替換為71mg,0.3mmol的3-溴-4-氯苯甲酸，得到84mg的產(chǎn)物，得率為66％。產(chǎn)物為白色固體。

¹H NMR(400MHz,CDCl₃)δ7.55(s,1H),7.18(d,J＝8.3Hz,1H),7.12(s,1H),7.10(d,J＝8.1Hz,2H),6.94(d,J＝8.1Hz,2H),5.78(s,1H),4.38(d,J＝7.2Hz,1H),3.46(s,3H),2.73(dt,J＝14.9,5.1Hz,1H),2.65–2.55(m,2H),2.52–2.31(m,4H),2.29–2.17(m,3H),2.08–1.92(m,3H),1.90(s,3H),1.80–1.65(m,2H),1.54–1.42(m,1H),1.39–1.28(m,1H),0.44(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.33,167.82,156.46,145.00,144.14,141.96,135.97,135.65,134.30,129.85,129.52,128.90,128.18,127.02,123.30,121.90,82.74,82.24,80.09,49.68,46.97,40.11,39.31,39.17,38.97,38.43,36.83,31.18,27.43,25.91,23.40,13.90,3.95.

HRMS(ESI-TOF):calcd for C₃₅H₃₆BrClNO₃[M+H]⁺632.1562；found 632.1559.

實(shí)施例28

2-Bromo-4-chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-037)

與實(shí)施例27不同之處在于：將3-溴-4-氯苯甲酸替換為2-溴-4-氯苯甲酸，得到68mg的產(chǎn)物，得率為54％。產(chǎn)物為白色固體，mp:128.8–129.9℃。

¹H NMR(400MHz,CDCl₃)δ7.37(s,1H),7.10–6.84(m,6H),5.77(s,1H),4.32(d,J＝7.0Hz,1H),3.48(s,3H),2.77–2.64(m,1H),2.63–2.54(m,2H),2.50–2.27(m,5H),2.26–2.12(m,3H),2.10–1.98(m,3H),1.97–1.92(m,1H),1.88(s,3H),1.79–1.60(m,2H),1.52–1.39(m,1H),1.36–1.24(m,1H),0.22(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.37,168.05,156.48,145.07,144.45,140.64,137.09,134.86,132.26,129.78,127.72,126.98,126.93,123.24,120.42,82.66,82.19,80.04,49.65,46.82,40.05,39.36,39.07,38.97,36.98,36.78,31.12,27.37,25.83,23.38,13.39,3.92.

實(shí)施例29

5-Chloro-2-fluoro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-038)

將105mg,0.6mmol的4-氯-2-氟苯甲酸、84mg,0.2mmol的美她司酮和192mg,1.0mmol的EDCI加入到2.0mL的CH2Cl2中，攪拌30小時(shí)，用方法C制備，PE/EtOAc＝2:1，得到70mg的產(chǎn)物，得率為61％。產(chǎn)物為白色固體，mp:121.5–122.7℃。

¹H NMR(400MHz,CDCl₃)δ7.23(d,J＝2.8Hz,1H),7.16–7.09(m,1H),7.03(d,J＝8.1Hz,2H),6.97(d,J＝8.0Hz,2H),6.69(t,J＝8.7Hz,1H),5.77(s,1H),4.34(d,J＝7.0Hz,1H),3.47(s,3H),2.75–2.64(m,1H),2.63–2.52(m,2H),2.49–2.29(m,4H),2.27–2.12(m,3H),2.09–1.98(m,2H),1.97–1.92(m,1H),1.89(s,3H),1.81–1.66(m,2H),1.53–1.40(m,1H),1.36–1.25(m,1H),0.31(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.36,165.18,157.71,156.49,155.23,145.17,144.44,140.66,130.95,130.87,129.78,129.31,129.13,127.72,127.08,126.78,126.59,123.23,116.99,116.76,82.71,82.26,80.07,49.67,46.85,40.15,39.39,39.19,38.92,37.36,36.82,31.15,27.41,25.82,23.40,13.44,3.89.

HRMS(ESI-TOF):calcd for C₃₅H₃₆ClFNO₃[M+H]⁺572.2362；found 572.2358.

實(shí)施例30

3-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-4-methoxy-N-methylbenzamide(FZU-0010-039)

與實(shí)施例28不同之處在于：將71mg,0.3mmol的2-溴-4-氯苯甲酸替換為56mg,0.3mmol的4-氯-3-甲氧基苯甲酸，得到86mg的產(chǎn)物，得率為73％，產(chǎn)物為白色固體，mp:122.1–123.4℃。

¹H NMR(400MHz,CDCl₃)δ7.30(s,1H),7.14(d,J＝8.5Hz,1H),7.07(d,J＝7.9Hz,2H),6.94(d,J＝7.7Hz,2H),6.62(d,J＝8.5Hz,1H),5.76(s,1H),4.37(d,1H),3.82(s,3H),3.46(s,3H),2.80–2.67(m,1H),2.63–2.52(m,2H),2.50–2.29(m,4H),2.28–2.17(m,3H),2.07–1.91(m,3H),1.89(s,3H),1.80–1.65(m,2H),1.54–1.41(m,1H),1.40–1.28(m,1H),0.46(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.42,168.73,156.57,155.99,145.23,143.64,142.64,131.31,129.76,129.31,128.57,128.01,126.96,123.20,121.60,110.63,82.67,82.25,80.05,56.18,49.64,46.94,40.10,39.33,39.22,38.92,38.47,36.79,31.17,27.41,25.84,23.40,13.91,3.91.

HRMS(ESI-TOF):calcd for C₃₆H₃₉ClNO₄[M+H]⁺584.2562；found 584.2559.

實(shí)施例31

4-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methyl-3-(trifluoromethyl)benzamide(FZU-0010-041)

與實(shí)施例30的不同之處在于：將56mg,0.3mmol的4-氯-3-甲氧基苯甲酸替換為67mg,0.3mmol的4-氯-3-三氟甲基苯甲酸，得到84mg的產(chǎn)物，得率為62％，產(chǎn)物為白色固體，mp:205.3–206.2℃。

¹H NMR(400MHz,CDCl₃)δ7.61(s,1H),7.38(d,J＝8.3Hz,1H),7.27(dd,J＝11.4,4.6Hz,1H),7.10(d,J＝8.2Hz,2H),6.94(d,J＝8.0Hz,2H),5.77(s,1H),4.37(d,J＝7.1Hz,1H),3.48(s,3H),2.71(dt,J＝14.9,5.2Hz,1H),2.62–2.52(m,2H),2.51–2.30(m,4H),2.26–2.17(m,3H),2.15–2.08(m,1H),2.07–1.92(m,2H),1.88(s,3H),1.79–1.62(m,2H),1.54–1.41(m,1H),1.38–1.26(m,1H),0.41(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.28,167.79,156.42,144.87,144.27,141.86,134.60,133.73,133.28,130.88,129.87,128.40,128.35,128.25,127.98,127.66,127.02,123.73,123.30,121.01,82.65,82.26,80.02,49.69,46.94,40.07,39.22,39.08,38.97,38.51,36.77,31.12,27.39,25.82,23.36,13.72,3.88.

HRMS(ESI-TOF):calcd for C₃₆H₃₆ClF₃NO₃[M+H]⁺622.2330；found 622.2323.

實(shí)施例32

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylthiazole-2-carboxamide(FZU-0010-048)

與實(shí)施例30的不同之處在于：將56mg,0.3mmol的4-氯-3-甲氧基苯甲酸替換為39mg,0.3mmol的噻唑-2-甲酸，得到70mg的產(chǎn)物，得率為66％，產(chǎn)物為白色固體，mp:194.8–195.9℃。

¹H NMR(400MHz,CDCl₃)δ7.63–7.43(m,1H),7.40–7.30(m,1H),7.22–6.98(m,4H),5.77(s,1H),4.43(d,J＝7.1Hz,1H),3.51(s,3H),2.77(dt,J＝10.5,5.7Hz,1H),2.63–2.54(m,2H),2.51–2.39(m,3H),2.38–2.34(m,1H),2.34–2.24(m,3H),2.24–2.18(m,2H),2.07–1.94(m,2H),1.88(s,3H),1.81–1.69(m,2H),1.54–1.42(m,1H),1.39–1.29(m,1H),0.52(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.42,161.12,156.59,145.48,144.37,143.00,141.64,129.67,127.91,127.18,123.11,82.58,82.32,80.02,49.71,46.98,40.20,39.30,39.17,38.88,36.81,31.11,27.34,25.85,23.37,13.86,3.89.

HRMS(ESI-TOF):calcd for C₃₂H₃₅N₂O₃S[M+H]⁺527.2363；found 527.2354.

實(shí)施例33

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylthiazole-5-carboxamide(FZU-0010-049)

與實(shí)施例32的不同之處在于：將噻唑-2-甲酸替換為噻唑-5-甲酸，PE/EtOAc＝1:2，得到78mg的產(chǎn)物，得率為74％，產(chǎn)物為白色固體，mp:123.5–124.7℃。

¹H NMR(400MHz,CDCl₃)δ8.65(s,1H),7.52(s,1H),7.27(d,J＝8.0Hz,2H),7.15(d,J＝7.8Hz,2H),5.79(s,1H),4.48(d,J＝7.1Hz,1H),3.43(s,3H),2.88–2.75(m,1H),2.66–2.56(m,2H),2.53–2.43(m,3H),2.41–2.36(m,1H),2.37–2.21(m,4H),2.17–2.08(m,1H),2.08–1.99(m,2H),1.98–1.93(m,1H),1.89(s,3H),1.81–1.68(m,2H),1.56–1.44(m,1H),1.40–1.31(m,1H),0.58(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.39,161.18,156.69,156.48,147.44,146.23,144.92,140.84,133.32,129.97,128.83,128.26,123.33,82.63,82.25,80.02,49.66,47.11,40.29,39.46,39.25,38.99,38.74,36.80,31.16,27.40,25.93,23.41,14.44,3.93.

HRMS(ESI-TOF):calcd for C₃₂H₃₅N₂O₃S[M+H]⁺527.2363；found 527.2355.

實(shí)施例34

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylpyrazine-2-carboxamide(FZU-0010-050)

與實(shí)施例33不同之處在于：將39mg,0.3mmol的噻唑-5-甲酸替換為50mg,0.4mmol的吡嗪-2-羧酸，得到72mg的產(chǎn)物，得率為69％，產(chǎn)物為白色固體，mp:116.5–117.7℃。

¹H NMR(400MHz,CDCl₃)δ8.69(s,1H),8.38(s,1H),8.21(s,1H),7.18–6.87(m,4H),5.77(s,1H),4.35(d,J＝6.5Hz,1H),3.52(s,3H),2.80–2.67(m,1H),2.66–2.51(m,2H),2.51–2.30(m,5H),2.26–2.13(m,3H),2.10–1.93(m,3H),1.84(s,3H),1.79–1.63(m,2H),1.52–1.40(m,1H),1.36–1.24(m,1H),0.36(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.39,166.49,156.52,149.87,145.13,144.91,144.50,144.11,142.83,141.29,129.74,127.92,127.15,123.20,82.66,82.24,79.98,49.66,46.90,40.04,39.27,39.12,38.89,37.87,36.79,31.10,27.33,25.83,23.36,13.71,3.90.

HRMS(ESI-TOF):calcd for C₃₃H₃₆N₃O₃[M+H]⁺522.2751；found 522.2741.

實(shí)施例35

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylthiophene-2-carboxamide(FZU-0010-051)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為51mg,0.4mmol的噻吩-2-羧酸，得到70mg的產(chǎn)物，得率為66％，產(chǎn)物為白色固體，mp:124.9–126.2℃。

¹H NMR(400MHz,CDCl₃)δ7.27(dd,J＝7.3,5.9Hz,1H),7.25–7.19(m,2H),7.16(d,J＝8.5Hz,2H),6.77–6.61(m,2H),5.79(s,1H),4.48(d,J＝7.2Hz,1H),3.43(s,3H),2.80(dt,J＝10.3,5.8Hz,1H),2.66–2.56(m,2H),2.53–2.42(m,3H),2.42–2.36(m,1H),2.34–2.16(m,4H),2.10–2.01(m,1H),1.99–1.94(m,1H),1.91(s,3H),1.83–1.69(m,2H),1.58–1.44(m,1H),1.41–1.33(m,1H),0.59(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.29,162.78,156.45,145.46,145.16,141.86,138.10,132.26,130.52,129.87,128.52,128.25,126.53,123.29,82.75,82.31,80.08,49.71,47.13,40.33,39.54,39.30,38.95,38.91,36.85,31.19,27.45,25.93,23.45,14.16,3.93.

實(shí)施例36

4-((4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)-4-oxobutanoic acid(FZU-0002-013)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為47mg，0.4mmol的丁二酸，根據(jù)方法c.得到100mg白色固體，收率67％。

¹H NMR(400MHz,CDCl₃)δ7.25(d,J＝8.2Hz,2H),7.13(d,J＝8.4Hz,2H),5.81(s,1H),4.46(d,J＝7.0Hz,1H),3.25(s,3H),2.88–2.75(m,1H),2.69–2.52(m,4H),2.51–2.37(m,4H),2.37–2.15(m,5H),2.10–1.92(m,2H),1.91(s,3H),1.81–1.67(m,2H),1.55–1.44(m,1H),1.39–1.32(m,1H),0.46(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.82,172.17,156.92,145.45,145.01,141.14,129.85,128.60,127.33,123.20,82.71,82.23,80.13,49.78,47.00,40.25,39.22,38.93,37.58,36.75,31.16,29.71,29.17,27.37,25.96,23.41,13.96,3.94.

HRMS(ESI-TOF):calcd for C₃₂H₃₆NO₅[M-H]^-514.2599；found 514.2576.

實(shí)施例37

2-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0002-017)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為56mg，0.36mmol鄰氯苯甲酸，根據(jù)方法c得到140mg白色固體，收率84％。

¹H NMR(400MHz,CDCl₃)δ7.48–7.33(m,1H),7.15(d,J＝7.8Hz,1H),7.10–7.04(m,2H),7.03–6.92(m,4H),5.76(s,1H),4.30(d,J＝7.2Hz,1H),3.49(s,3H),2.66(dt,J＝14.9,5.4Hz,1H),2.60–2.53(m,2H),2.47–2.26(m,4H),2.25–2.09(m,3H),2.02–1.97(m,2H),1.88(s,3H),1.77–1.64(m,2H),1.49–1.38(m,1H),1.32–1.27(m,1H),0.22(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.40,168.28,156.51,145.30,144.19,140.87,136.53,130.65,129.70,129.36,128.83,127.56,126.95,126.10,123.16,82.65,82.24,80.04,49.61,46.77,40.09,39.33,39.15,38.87,38.69,36.92,36.79,31.12,27.37,25.76,23.37,13.38,3.89.

HRMS(ESI-TOF):calcd for C₃₅H₃₅ClNO₃[M-H]^-552.2311；found 552.2294.

實(shí)施例38

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-4-methoxy-N-methylbenzamide(FZU-0010-016)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為63mg，0.36mmol對(duì)甲氧基苯甲酸，根據(jù)方法c得到120mg白色固體，收率70％，mp:114.2–115.6℃。

¹H NMR(400MHz,CDCl₃)δ7.21(d,J＝8.4Hz,2H),7.05(d,J＝8.0Hz,2H),6.94(d,J＝8.1Hz,2H),6.60(d,J＝8.5Hz,2H),5.77(s,1H),4.37(d,J＝6.9Hz,1H),3.72(s,3H),3.46(s,3H),2.83–2.70(m,1H),2.63–2.53(m,2H),2.50–2.33(m,2H),2.31–2.18(m,2H),2.16–2.10(m,1H),2.08–1.94(m,3H),1.90(s,3H),1.79–1.67(m,2H),1.50–1.42(m,1H),1.39–1.30(m,1H),0.42(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.45,170.12,160.62,156.60,145.38,143.26,143.10,131.09,129.71,127.86,127.77,127.00,123.18,112.84,82.70,82.28,80.07,55.28,49.70,46.97,40.11,39.40,39.21,38.92,38.45,36.81,31.16,27.41,25.85,23.42,13.84,3.92.

HRMS(ESI-TOF):calcd for C₃₆H₄₀NO₄[M+H]⁺550.2952；found 550.2950.

實(shí)施例39

4-Chloro-2,5-difluoro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-018)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為82mg，0.2mmol4-氯-2,5-二氟苯甲酸，根據(jù)方法c得到55mg白色固體，收率46％，mp:117.1–118.3℃。

¹H NMR(400MHz,CDCl₃)δ7.11–7.00(m,3H),6.96(d,J＝7.9Hz,2H),6.86–6.74(m,1H),5.78(s,1H),4.36(d,J＝7.0Hz,1H),3.46(s,3H),2.71(dt,J＝10.8,5.2Hz,1H),2.60–2.54(m,2H),2.49–2.29(m,4H),2.25–2.12(m,4H),2.01–1.91(m,2H),1.87(s,4H),1.80–1.67(m,2H),1.52–1.39(m,1H),1.36–1.28(m,1H),0.29(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.31,164.45,156.44,145.00,144.65,140.47,129.84,127.81,127.08,123.29,117.88,117.61,116.90,116.65,82.69,82.24,80.04,49.71,46.89,40.11,39.42,39.12,38.97,37.37,36.80,31.14,27.40,25.87,23.39,13.35,3.90.

HRMS(ESI-TOF):calcd for C₃₅H₃₅ClF₂NO₃[M+H]⁺590.2268；found 590.2245.

實(shí)施例40

2,5-Dichloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-020)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為152mg，0.08mmol的2,6-二氯苯甲酸，根據(jù)方法c.得到99mg黃色油狀物，收率83％。

¹H NMR(400MHz,CDCl₃)δ7.22–6.94(m,7H),5.78(s,1H),4.34(d,J＝7.3Hz,1H),3.50(s,3H),2.69(dt,J＝15.0,5.4Hz,1H),2.63–2.57(m,2H),2.51–2.30(m,4H),2.29–2.19(m,2H),2.18–2.09(m,2H),2.08–2.00(m,1H),1.97(s,1H),1.91(s,3H),1.79–1.62(m,2H),1.52–1.42(m,1H),1.39–1.30(m,1H),0.28(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.35,166.77,156.42,145.08,144.65,140.57,137.93,132.26,130.67,129.87,129.78,129.20,128.82,127.84,126.96,123.32,82.83,82.23,80.15,49.67,46.81,40.16,39.40,39.21,38.96,37.01,36.85,31.18,27.45,25.86,23.43,13.31,3.94.

HRMS(ESI-TOF):calcd for C₃₅H₃₆Cl₂NO₃[M+H]⁺588.2067；found 588.2058.

實(shí)施例41

5-Chloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N,2-dimethylbenzamide(FZU-0010-025)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為51mg，0.3mmol的5-氯-2-甲基苯甲酸，得到63mg白色固體，收率55％，mp:125.4–126.8℃。

¹H NMR(400MHz,CDCl₃)δ7.13–6.98(m,3H),6.94–6.75(m,3H),5.77(s,1H),4.32(d,J＝5.6Hz,1H),3.49(s,3H),2.78–2.62(m,1H),2.62–2.54(m,2H),2.49–2.31(m,4H),2.27(s,3H),2.24–2.10(m,3H),2.07–1.95(m,3H),1.88(s,3H),1.79–1.63(m,2H),1.53–1.40(m,1H),1.37–1.25(m,1H),0.32(s,3H).

HRMS(ESI-TOF):calcd for C₃₆H₃₉ClNO₃[M+H]⁺568.2613；found 568.2598.

實(shí)施例42

4-Chloro-2-fluoro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-031)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為70mg，0.4mmol的4-氯-2-氟苯甲酸，根據(jù)方法c.得到38mg白色固體，收率33％，mp:125.3–126.5℃℃。

¹H NMR(400MHz,CDCl₃)δ7.28(s,1H),7.23–7.14(m,1H),7.01(d,J＝8.6Hz,2H),6.96(d,J＝7.0Hz,2H),6.84–6.71(m,1H),5.78(d,J＝8.5Hz,1H),4.46–4.28(m,1H),3.62–3.36(m,3H),2.77–2.64(m,1H),2.62–2.52(m,2H),2.49–2.32(m,3H),2.26–2.11(m,3H),2.06–1.99(m,1H),1.89(s,3H),1.82–1.73(m,2H),1.52–1.40(m,1H),1.34–1.22(m,2H),0.93–0.76(m,2H),0.26(s,3H).

HRMS(ESI-TOF):calcd for C₃₅H₃₅ClFNO₃[M+H]⁺572.2362；found 527.2356.

實(shí)施例43

2,4-Dichloro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-033)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為76mg，0.4mmol的2,4-二氯苯甲酸，根據(jù)方法c.得到71mg白色固體，收率60％，mp:121.5–122.6℃℃。

¹H NMR(400MHz,CDCl₃)δ7.37(s,1H),7.19(s,1H),7.02–6.98(m,5H),5.78(s,1H),4.33(s,1H),3.49(s,3H),2.77–2.64(m,1H),2.57–2.53(m,2H),2.49–2.28(m,4H),2.25–2.09(m,3H),2.08–1.98(m,1H),1.89(s,3H),1.82–1.66(m,3H),1.51–1.41(m,1H),1.32–1.21(m,1H),0.94–0.85(m,1H),0.23(s,3H).

HRMS(ESI-TOF):calcd for C₃₅H₃₆Cl₂NO₃[M+H]⁺588.2067；found 588.2059.

實(shí)施例44

3-Chloro-2-fluoro-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-035)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為52mg，0.3mmol的2-氟，3-氯苯甲酸，根據(jù)方法c.得到130mg白色固體，收率80％，mp:123.5–124.5℃。

¹H NMR(400MHz,CDCl₃)δ7.23(t,J＝7.1Hz,1H),7.19–7.11(m,1H),7.10–6.88(m,5H),5.77(s,1H),4.33(d,J＝7.0Hz,1H),3.49(s,3H),2.77–2.63(m,1H),2.62–2.52(m,2H),2.48–2.31(m,4H),2.24–2.10(m,3H),2.06–1.92(m,3H),1.89(s,3H),1.76–1.62(m,2H),1.52–1.40(m,1H),1.39–1.28(m,1H),0.32(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.40,165.61,156.49,145.17,144.45,140.69,131.46,129.82,127.72,127.15,126.86,126.68,124.52,123.27,82.76,82.27,80.14,49.67,46.84,40.17,39.34,39.22,38.96,37.37,36.84,31.18,27.44,25.83,23.42,13.46,3.91.

HRMS(ESI-TOF):calcd for C₃₅H₃₅ClFNO₃[M+H]⁺572.2362；found 527.2359.

實(shí)施例45

4-(Dimethylamino)-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-042)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為66mg，0.4mmol的4-二甲氨基苯甲酸，根據(jù)方法c.獲得86mg白色固體，收率76％，mp:122.7–123.7℃。

¹H NMR(400MHz,CDCl₃)δ7.17(d,J＝8.9Hz,2H),7.06(d,J＝8.0Hz,2H),6.97(d,J＝8.4Hz,2H),6.36(d,J＝8.9Hz,2H),5.76(s,1H),4.38(s,1H),3.47(s,3H),2.90(s,6H),2.78–2.70(m,1H),2.61–2.53(m,2H),2.49–2.34(m,4H),2.28–2.17(m,3H),2.07–1.97(m,2H),1.90(s,3H),1.80–1.65(m,2H),1.54–1.41(m,1H),1.38–1.29(m,1H),0.91–0.86(m,1H),0.51(s,3H).

HRMS(ESI-TOF):calcd for C₃₇H₄₃N₂O₃[M+H]⁺563.3268；found 563.3261.

實(shí)施例46

3-(Dimethylamino)-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0010-043)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為66mg，0.4mmol的3-二甲氨基苯甲酸，根據(jù)方法c.獲得57mg白色固體，收率50％，mp:122.4–123.4℃。

¹H NMR(400MHz,CDCl₃)δ7.02–7.00(m,2H),6.98–6.96(s,2H),6.88(d,J＝6.2Hz,1H),6.74(s,1H),6.56(d,J＝4.4Hz,1H),6.44(d,J＝5.4Hz,1H),5.75(s,1H),4.33(s,1H),4.12(d,J＝6.8Hz,1H),3.47(s,3H),2.82(s,6H),2.72–2.68(m,1H),2.58–2.55(m,2H),2.37–2.34(m,4H),2.19–2.13(m,3H),1.99–1.96(m,1H),1.88(s,3H),1.73–1.70(m,2H),1.45–1.42(m,1H),1.32–1.28(m,1H),0.87–0.84(m,1H),0.42(s,3H).

HRMS(ESI-TOF):calcd for C₃₇H₄₃N₂O₃[M+H]⁺563.3268；found 563.3260.

實(shí)施例47

(E)-3-(4-(dimethylamino)phenyl)-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylacrylamide(FZU-0010-044)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為57mg，0.3mmol的4-二甲基氨基肉桂酸，根據(jù)方法c獲得66mg黃色油狀物，收率55％。

¹H NMR(400MHz,CDCl₃)δ7.56(d,J＝15.5Hz,1H),7.23(s,2H),7.14(d,J＝8.2Hz,3H),6.57(d,J＝7.0Hz,2H),6.06(d,J＝15.5Hz,1H),5.79(s,1H),4.49(d,J＝6.6Hz,1H),3.37(s,3H),2.96(s,6H),2.81(d,J＝7.8Hz,1H),2.60(s,2H),2.55–2.20(m,8H),2.11–1.99(m,2H),1.91(s,3H),1.81–1.69(m,3H),1.53–1.46(m,1H),1.41–1.35(m,1H),0.60(s,3H).

HRMS(ESI-TOF):calcd for C₃₉H₄₅N₂O₃[M+H]⁺589.3425；found 589.3414.

實(shí)施例48

2-Hydroxy-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methyl-2-phenylacetamide(FZU-0010-052)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為61mg，0.4mmol的扁桃酸，根據(jù)方法c得到45mg白色固體，收率41％，mp:122.1–123.5℃。

¹H NMR(400MHz,CDCl₃)δ7.65–7.34(m,2H),7.23–7.05(m,3H),6.98–6.86(m,1H),6.85–6.69(m,2H),5.80(d,J＝10.2Hz,1H),5.02(s,1H),4.58–4.30(m,1H),3.33–3.20(m,2H),2.86–2.70(m,1H),2.67–2.56(m,1H),2.53–2.36(m,4H),2.34–2.21(m,3H),2.12–1.96(m,3H),1.90(s,3H),1.79–1.69(m,2H),1.54–1.33(m,2H),1.31–1.21(m,1H),0.94–0.83(m,1H),0.60(s,3H).

HRMS(ESI-TOF):calcd for C₃₆H₄₀NO₄[M+H]⁺550.2952；found 550.2945.

實(shí)施例49

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methyl-2-naphthamide(FZU-0025-003)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為42mg，0.24mmol的2-萘甲酸，根據(jù)方法c得到87mg白色固體，收率77％，,mp:129.3–130.5℃。¹H NMR(400MHz,CDCl₃)δ7.85(s,1H),7.72(d,J＝7.9Hz,1H),7.67(d,J＝7.8Hz,1H),7.53(d,J＝8.5Hz,1H),7.49–7.37(m,2H),7.27(d,J＝9.1Hz,1H),6.98(q,J＝8.5Hz,4H),5.73(s,1H),4.31(d,J＝7.2Hz,1H),3.55(s,3H),2.65(dt,J＝14.8,5.1Hz,1H),2.57–2.46(m,2H),2.45–2.20(m,5H),2.20–2.08(m,3H),2.02–1.92(m,2H),1.88(s,3H),1.76–1.62(m,2H),1.47–1.37(m,1H),1.30–1.24(m,1H),0.35(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.37,170.41,156.53,145.25,143.49,142.59,133.57,133.08,132.30,129.63,128.60,127.87,127.63,127.27,127.09,127.02,126.34,125.63,123.11,82.61,82.24,80.03,49.58,46.87,40.08,39.28,39.17,38.86,38.39,36.75,31.12,27.37,25.76,23.33,13.66,3.90.HRMS(ESI-TOF):calcd for C₃₉H₄₀NO₃[M+H]⁺570.3003；found 570.2968.

實(shí)施例50

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methyl-[1,1'-biphenyl]-3-carboxamide(FZU-0025-004)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為48mg，0.24mmol的3-苯基苯甲酸，根據(jù)方法c得到90mg白色固體，收率76％，mp:126.9–127.9℃。

¹H NMR(400MHz,CDCl₃)δ7.49–7.25(m,7H),7.24–7.13(m,2H),7.05(d,J＝8.2Hz,2H),7.02–6.93(m,2H),5.75(s,1H),4.33(d,J＝7.1Hz,1H),3.52(s,3H),2.65(dt,J＝15.0,5.2Hz,1H),2.58–2.47(m,2H),2.43–2.30(m,3H),2.28–2.08(m,4H),2.03–1.95(m,2H),1.89(s,3H),1.77–1.63(m,2H),1.50–1.40(m,1H),1.30–1.26(m,1H),0.27(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.40,170.51,156.54,145.21,143.43,142.62,140.34,140.11,136.34,129.70,128.83,128.24,128.21,127.85,127.79,127.62,127.49,127.09,126.93,123.18,82.63,82.16,80.09,49.65,46.85,40.07,39.16,38.94,38.33,36.76,31.14,27.38,25.80,23.34,22.71,13.65,3.91.

HRMS(ESI-TOF):calcd for C₄₁H₄₂NO₃[M+H]⁺596.3159；found 596.3146.

實(shí)施例51

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N,3,4-trimethylbenzamide(FZU-0025-005)

與實(shí)施例34不同之處在于：將50mg,0.4mmol的吡嗪-2-羧酸替換為36mg，0.24mmol的3,4-二甲基苯甲酸，根據(jù)方法c得到84mg白色固體，收率77％，mp:131.6–132.5℃。

¹H NMR(400MHz,CDCl₃)δ7.11(s,1H),7.04(d,J＝8.2Hz,2H),6.94(d,J＝8.4Hz,2H),6.91(s,1H),6.83(d,J＝7.8Hz,1H),5.77(s,1H),4.36(d,J＝7.2Hz,1H),3.45(s,3H),2.72(dt,J＝10.8,5.2Hz,1H),2.62–2.54(m,2H),2.50–2.34(m,3H),2.27–2.17(m,3H),2.15(s,3H),2.10(s,3H),2.06–1.95(m,3H),1.90(s,3H),1.80–1.65(m,2H),1.57–1.43(m,1H),1.38–1.30(m,1H),0.45(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.33,170.55,156.51,145.37,143.10,142.90,138.54,135.83,133.01,130.23,129.58,128.63,127.69,126.86,126.56,123.06,82.59,82.23,79.98,49.58,46.85,40.06,39.26,39.17,38.83,38.35,36.74,31.10,27.35,25.75,23.34,22.63,19.65,13.62,3.84.

HRMS(ESI-TOF):calcd for C₃₇H₄₂NO₃[M+H]⁺548.3159；found 548.3152.

實(shí)施例52

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide(FZU-0025-001)

將48mg,0.24mmol的4'-羥基聯(lián)苯基-4-羧酸，83mg,0.2mmol的美她司酮和115mg,0.6mmol的EDCI加入到2.0mL的CH₂Cl₂中，PE/EtOAc＝1:1，按方法C制備得到87mg的產(chǎn)物，得率為73％，產(chǎn)物為白色固體，mp:125.5–126.5℃。

¹H NMR(400MHz,CDCl₃)δ7.48(d,J＝7.6Hz,2H),7.41(t,J＝7.5Hz,2H),7.37–7.29(m,5H),7.06(d,J＝8.2Hz,2H),7.01–6.93(m,2H),5.77(s,1H),4.37(d,J＝7.0Hz,1H),3.50(s,3H),2.74(dt,J＝14.8,5.0Hz,1H),2.62–2.52(m,2H),2.51–2.32(m,4H),2.28–2.18(m,3H),2.15–2.08(m,1H),2.04–1.95(m,2H),1.90(s,3H),1.80–1.65(m,2H),1.51–1.39(m,1H),1.36–1.26(m,1H),0.42(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.37,170.30,156.58,145.25,143.46,142.62,142.32,140.08,134.55,129.71,129.45,128.92,127.85,127.07,126.27,123.17,82.65,82.24,80.02,49.70,46.98,40.08,39.39,39.17,38.95,38.28,36.80,31.15,27.40,25.86,23.39,14.00,3.91.

HRMS(ESI-TOF):calcd for C₄₁H₄₂NO₃[M+H]⁺596.3159；found 539.3146.

實(shí)施例53

4-(tert-Butyl)-N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylbenzamide(FZU-0025-002)

與實(shí)施例52不同之處在于：將48mg,0.24mmol的4'-羥基聯(lián)苯基-4-羧酸替換為43mg,0.24mmol的對(duì)叔丁基苯甲酸，得到74mg的產(chǎn)物，得率為64％，產(chǎn)物為白色固體，mp:216.9-217.8℃。

¹H NMR(400MHz,CDCl₃)δ7.16(d,J＝8.2Hz,2H),7.11(d,J＝8.2Hz,2H),7.03(d,J＝8.1Hz,2H),6.94(d,J＝8.1Hz,2H),5.75(s,1H),4.36(d,J＝7.0Hz,1H),3.48(s,3H),2.73(dt,J＝14.7,5.1Hz,1H),2.63–2.51(m,2H),2.51–2.32(m,4H),2.26–2.16(m,3H),2.06–1.89(m,3H),1.88(s,3H),1.79–1.63(m,2H),1.53–1.39(m,1H),1.35–1.27(m,1H),1.20(s,9H),0.40(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.30,170.69,156.52,152.84,145.32,143.17,142.83,132.87,129.67,128.75,127.73,127.10,124.53,123.17,82.64,82.30,80.01,49.70,46.96,40.06,39.33,39.17,38.95,38.16,36.85,34.71,31.17,27.39,25.84,23.43,13.96,3.93.

HRMS(ESI-TOF):calcd for C₃₉H₄₆NO₃[M+H]⁺576.3472；found 576.3462.

實(shí)施例54

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-4-isopropyl-N-methylbenzamide(FZU-0025-006)

與實(shí)施例52不同之處在于：將48mg,0.24mmol的4'-羥基聯(lián)苯基-4-羧酸替換為39mg,0.24

mmol的4-異丙基苯甲酸，得到82mg的產(chǎn)物，得率為73％，產(chǎn)物為白色固體，mp:237.9-238.9℃。

¹H NMR(400MHz,CDCl₃)δ7.16(d,J＝7.4Hz,2H),7.03(d,J＝7.5Hz,2H),6.99–6.86(m,4H),5.76(s,1H),4.36(d,J＝6.0Hz,1H),3.47(s,3H),2.87–2.69(m,2H),2.63–2.48(m,2H),2.48–2.29(m,4H),2.27–2.16(m,3H),2.14–2.06(m,1H),2.06–1.94(m,2H),1.88(s,3H),1.78–1.62(m,2H),1.51–1.38(m,1H),1.35–1.26(m,1H),1.13(d,J＝6.5Hz,6H),0.37(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.33,170.70,156.54,150.63,145.36,143.21,142.83,133.23,129.66,129.03,127.74,127.08,125.66,123.15,82.63,82.30,80.01,49.67,46.95,40.07,39.33,39.17,38.92,38.18,36.82,33.92,31.14,27.39,25.82,23.80,23.76,23.42,13.87,3.91.

HRMS(ESI-TOF):calcd for C₃₈H₄₄NO₃[M+H]⁺562.3316；found 562.3306.

實(shí)施例55

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methyl-2-phenylacetamide(FZU-0025-012)

與實(shí)施例52不同之處在于：將48mg,0.24mmol的4'-羥基聯(lián)苯基-4-羧酸替換為33mg,0.24mmol的苯乙酸，得到83mg的產(chǎn)物，得率為78％，產(chǎn)物為白色固體，mp:126.7-127.5℃。

¹H NMR(400MHz,CDCl₃)δ7.26–7.12(m,5H),7.11–6.85(m,4H),5.80(s,1H),4.46(d,J＝7.0Hz,1H),3.46(s,2H),3.24(d,J＝8.5Hz,3H),2.88–2.74(m,1H),2.68–2.57(m,2H),2.54–2.38(m,4H),2.36–2.17(m,4H),2.13–1.98(m,2H),1.92(s,3H),1.84–1.71(m,2H),1.59–1.45(m,1H),1.41–1.31(m,1H),0.57(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.37,171.15,156.55,145.21,144.79,141.59,135.38,129.82,129.00,128.34,128.28,127.72,126.63,123.25,82.70,82.28,80.03,49.72,47.00,41.11,40.22,39.27,39.22,38.96,37.68,36.84,31.14,27.40,25.97,23.43,14.01,3.94.

HRMS(ESI-TOF):calcd for C₃₆H₄₀NO₃[M+H]⁺534.3003；found 534.2997.

實(shí)施例56

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N,3-dimethylbenzamide(FZU-0025-014)

與實(shí)施例52不同之處在于：將48mg,0.24mmol的4'-羥基聯(lián)苯基-4-羧酸替換為33mg,0.24mmol的3-甲基苯甲酸，得到74mg的產(chǎn)物，得率為69％，產(chǎn)物為白色固體，mp:128.6-129.5℃。

¹H NMR(400MHz,CDCl₃)δ7.15(s,1H),7.07–6.97(m,3H),6.98–6.84(m,4H),5.76(s,1H),4.35(d,J＝7.1Hz,1H),3.46(s,3H),2.71(dt,J＝14.8,5.2Hz,1H),2.63–2.52(m,2H),2.51–2.28(m,4H),2.26–2.20(m,2H),2.19(s,3H),2.15–2.08(m,1H),2.07–1.93(m,3H),1.90(s,3H),1.79–1.66(m,2H),1.53–1.42(m,1H),1.37–1.28(m,1H),0.42(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.44,170.72,156.59,145.38,143.25,142.71,137.41,135.65,130.44,129.66,129.56,127.74,127.36,126.98,125.99,123.14,82.67,82.27,80.06,49.67,46.92,40.09,39.27,39.19,38.90,38.30,36.80,31.15,27.40,25.81,23.40,21.33,13.72,3.92.

HRMS(ESI-TOF):calcd for C₃₆H₄₀NO₃[M+H]⁺534.3003；found 534.2996.

實(shí)施例57

N-(4-((8S,11R,13S,14S,17S)-17-hydroxy-13-methyl-3-oxo-17-(prop-1-yn-1-yl)-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N,4-dimethylbenzamide(FZU-0025-015)

與實(shí)施例52不同之處在于：將48mg,0.24mmol的4'-羥基聯(lián)苯基-4-羧酸替換為33mg,0.24mmol的對(duì)甲苯甲酸，得到73mg的產(chǎn)物，得率為68％，產(chǎn)物為白色固體，mp:125.7-126.8℃。

¹H NMR(400MHz,CDCl₃)δ7.20–7.09(m,2H),7.08–6.98(m,2H),6.98–6.80(m,4H),5.77(s,1H),4.37(d,J＝7.1Hz,1H),3.48(s,3H),2.81–2.68(m,1H),2.65–2.50(m,2H),2.50–2.32(m,4H),2.31–2.17(m,6H),2.16–2.10(m,1H),2.06–1.95(m,2H),1.89(s,3H),1.81–1.67(m,2H),1.52–1.40(m,1H),1.39–1.28(m,1H),0.43(s,3H).

¹³C NMR(101MHz,CDCl₃)δ199.44,170.59,156.58,145.43,143.40,142.85,139.90,132.78,129.68,129.05,128.27,127.82,127.04,123.15,82.69,82.29,80.06,49.65,46.95,40.13,39.40,39.23,38.91,38.32,36.81,31.16,27.41,25.83,23.42,21.42,13.72,3.92.

HRMS(ESI-TOF):calcd for C₃₆H₄₀NO₃[M+H]⁺534.3003；found 534.3000.

實(shí)施例58米非司酮衍生物對(duì)三陰性乳腺癌細(xì)胞的作用

以人三陰性乳腺癌細(xì)胞系HCC1937和SUM149PT為測(cè)試細(xì)胞系(細(xì)胞購(gòu)自美國(guó)模式培養(yǎng)物集存庫(kù)，ATCC)。

細(xì)胞培養(yǎng)方法：取出液氮中凍存的HCC1937細(xì)胞，在37℃的溫水中解凍，將細(xì)胞凍存管置于離心機(jī)中，500g離心5min，棄去上清液，加入1mL RPMI-1640完全培養(yǎng)液，輕輕吹打均勻，將細(xì)胞懸液加入培養(yǎng)皿中，補(bǔ)加9mL RPMI-1640完全培養(yǎng)液，將培養(yǎng)皿置于5％CO2、37℃培養(yǎng)箱中培養(yǎng)。取出液氮中凍存的SUM149PT細(xì)胞，在37℃的溫水中解凍，將細(xì)胞凍存管置于離心機(jī)中，500g離心5min，棄去上清液，加入1mL F-12完全培養(yǎng)液，輕輕吹打均勻，將細(xì)胞懸液加入培養(yǎng)皿中，補(bǔ)加9mL F-12完全培養(yǎng)液，將培養(yǎng)皿置于5％CO2、37℃培養(yǎng)箱中培養(yǎng)。細(xì)胞毒性實(shí)驗(yàn)：將HCC1937或SUM149PT細(xì)胞以3×10⁴個(gè)細(xì)胞/孔的密度接種到48孔培養(yǎng)板中，培養(yǎng)24h后，更換培養(yǎng)液為新鮮血清培養(yǎng)液，加入0、5μM、10μM、15μM、20μM五個(gè)濃度的米非司酮或用本發(fā)明方法制備所得的米非司酮系列衍生物，其中FZU-0000系列3種，F(xiàn)ZU-0002系列10種，F(xiàn)ZU-0010系列39種，F(xiàn)ZU-0025系列13種，孵育48h后，吸棄孔中溶液，每孔加入200μl 10％三氯醋酸(TCA)，室溫放置1小時(shí)或4℃過(guò)夜固定；固定好的細(xì)胞吸棄固定液，以蒸餾水清洗5遍，室溫晾干；每孔加入200μl 0.4％(w/v)的SRB染液(1％的乙酸配制)，室溫染色30min后倒掉染液，用1％(v/v)乙酸沖洗5次，去除未結(jié)合的染料，室溫晾干后每孔加入200μl非緩沖Tris-base堿液(10mM，pH10.5)溶解與細(xì)胞蛋白結(jié)合的染料，水平搖床上振蕩20min，采用酶標(biāo)儀530nm處測(cè)定光吸收值(A)，按照以下公式計(jì)算：細(xì)胞存活率％＝(試驗(yàn)組平均A值/空白對(duì)照組平均A值)×100％。結(jié)果如表1所示，結(jié)果顯示，HCC1937和SUM149PT細(xì)胞的存活率與米非司酮衍生物化合物濃度呈顯著相關(guān)性，化合物濃度越高，存活率越低，也表明部分米非司酮衍生物較米非司酮抗腫瘤活性強(qiáng)，濃度低

表1：米非司酮衍生物對(duì)三陰性乳腺癌細(xì)胞擴(kuò)增的影響

^a Triple-negative breast cancer(TNBC)cell lines:HCC1937and SUM149PT.Software:MasterPlex ReaderFit 2010,MiraiBio,Inc.The values are the mean±SE of at least three independent experiments.^b If a specific compound is given value>20,it indicates that a specific IC₅₀cannot be calculated from the data points collected,meaning“no effect”.NT代表未經(jīng)測(cè)試(No Test)。