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CN103980177A - Acetylene compound as well as preparation method and application thereof - Google Patents

Acetylene compound as well as preparation method and application thereof Download PDF

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CN103980177A
CN103980177A CN201410211924.6A CN201410211924A CN103980177A CN 103980177 A CN103980177 A CN 103980177A CN 201410211924 A CN201410211924 A CN 201410211924A CN 103980177 A CN103980177 A CN 103980177A
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enediyne
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dna
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CN103980177B (en
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胡爱国
宋德朋
孙士元
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East China University of Science and Technology
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    • C07ORGANIC CHEMISTRY
    • C07DHETEROCYCLIC COMPOUNDS
    • C07D207/00Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D207/02Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/44Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members
    • C07D207/444Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members having two doubly-bound oxygen atoms directly attached in positions 2 and 5
    • C07D207/448Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members having two doubly-bound oxygen atoms directly attached in positions 2 and 5 with only hydrogen atoms or radicals containing only hydrogen and carbon atoms directly attached to other ring carbon atoms, e.g. maleimide

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Abstract

本发明涉及一种可用于抗癌药物的新型烯二炔化合物及其制备方法和应用,本发明提出一种烯二炔化合物,其中,R代表官能基团;R=R、CR1R2R3、(CR1R2)nX或NR1R2;其中R,R1,R2,R3为氢、卤素、C1-C24烷基链、芳香基团、杂环基团;n为1~6;X为氨基、醚基、酰胺基、巯基、膦基、卤素或酯基。所述芳香基团是苯基、萘基、芘基、蒽基、烷基或卤素;所述杂环基团是吡咯、噻吩、吡啶、哌嗪、吡喃、呋喃、咪唑、吡唑或吗啡啉。经实验证明,这类烯二炔可以导致DNA明显的链断裂以及造成肿瘤细胞的凋亡,说明合成的烯二炔具有较高的生物活性。

The present invention relates to a novel enediyne compound that can be used in anticancer drugs and its preparation method and application. The present invention provides an enediyne compound, Among them, R represents a functional group; R=R, CR 1 R 2 R 3 , (CR 1 R 2 ) n X or NR 1 R 2 ; where R, R 1 , R 2 , R 3 are hydrogen, halogen, C1 -C24 alkyl chain, aromatic group, heterocyclic group; n is 1~6; X is amino, ether, amido, mercapto, phosphino, halogen or ester. The aromatic group is phenyl, naphthyl, pyrenyl, anthracenyl, alkyl or halogen; the heterocyclic group is pyrrole, thiophene, pyridine, piperazine, pyran, furan, imidazole, pyrazole or morphine phylloline. It has been proved by experiments that this kind of enediyne can cause obvious strand breaks of DNA and cause apoptosis of tumor cells, indicating that the synthesized enediyne has high biological activity.

Description

一种烯二炔化合物及其制备方法和应用A kind of enediyne compound and its preparation method and application

技术领域 technical field

本发明涉及一种可用于抗癌药物的新型烯二炔化合物及其制备方法和应用。 The invention relates to a novel enediyne compound which can be used for anticancer drugs, its preparation method and application.

背景技术 Background technique

癌症是导致人类死亡的重大疾病之一,临床上用于治疗癌症的方法有手术治疗,放疗和化疗等,以杀死恶性肿瘤细胞,其中化学治疗是主要手段。常用的治疗药物主要是抗生素之类,比如阿霉素,多柔比星,紫杉醇,卡铂等。这些抗生素类药物在治疗癌症的同时本身也带来很大的毒副作用。因此,寻找低毒,高效,高选择性的新型抗癌药物一直是科学家面临的巨大挑战。 Cancer is one of the major diseases that cause human death. Clinically, the methods used to treat cancer include surgery, radiotherapy and chemotherapy to kill malignant tumor cells, among which chemotherapy is the main method. Commonly used therapeutic drugs are mainly antibiotics, such as doxorubicin, doxorubicin, paclitaxel, carboplatin, etc. These antibiotic drugs themselves also bring great toxic and side effects while treating cancer. Therefore, finding new anticancer drugs with low toxicity, high efficiency and high selectivity has always been a great challenge for scientists.

烯二炔类化合物,作为一种新颖的抗肿瘤抗生素,具有独特的分子结构和抗肿瘤机制以及高效的杀伤细胞作用,是目前为止活性最强的抗肿瘤抗生素。这类化合物通过Bergman Cyclization反应可以生成苯基双自由基,形成的自由基夺取DNA链上的H原子,从而导致DNA的单链或者双链断裂。一些天然的烯二炔抗肿瘤抗生素如Dynemicins,Calicheamicins,Esperamcins,C1027等,其抗癌活性比阿霉素高1000倍以上。这些天然的烯二炔大都是从微生物的代谢中分离得到的。人工合成的烯二炔因其发生Bergman Cyclization反应的条件相比天然烯二炔来说比较苛刻,限制了在生物学上的应用。此外,一些天然的抗癌药物也具有很强的副毒性,如何提高抗癌药物的选择性以此来降低副作用也是一个需要解决的问题。 Enediynes, as a novel anti-tumor antibiotic, have a unique molecular structure, anti-tumor mechanism and high-efficiency killing cells, and are by far the most active anti-tumor antibiotics. This type of compound can generate phenyl diradicals through Bergman Cyclization reaction, and the formed free radicals capture the H atoms on the DNA chain, resulting in single-strand or double-strand breaks of DNA. Some natural enediyne antitumor antibiotics, such as Dynemicins, Calicheamicins, Esperamcins, C1027, etc., have anticancer activity more than 1000 times higher than that of doxorubicin. Most of these natural enediynes are isolated from the metabolism of microorganisms. Synthetic enediynes are more harsh than natural enediynes due to the Bergman Cyclization reaction conditions, which limits their biological applications. In addition, some natural anticancer drugs also have strong side effects. How to improve the selectivity of anticancer drugs to reduce side effects is also a problem that needs to be solved.

上述是烯二炔类抗肿瘤抗生素抗癌机理。 The above is the anticancer mechanism of enediynes antitumor antibiotics.

关于合成可用于抗癌药物方面的烯二炔的工作从1992年就有所研究。除了合成天然烯二炔抗生素的同系物外,人工合成的烯二炔一部分集中在光致引发反应,这些反应需要紫外光的照射;一部分主要形成金属配合物,但局限性较大。虽然部分烯二炔可以在温和条件下发生反应,但不具有对肿瘤细胞和正常细胞的选择性,这些都限制了烯二炔在生物抗肿瘤方面的应用。 The work on the synthesis of enediynes that can be used in anticancer drugs has been studied since 1992. In addition to synthesizing homologues of natural enediyne antibiotics, some of the artificially synthesized enediynes focus on photoinitiated reactions, which require ultraviolet light irradiation; some mainly form metal complexes, but have relatively large limitations. Although some enediynes can react under mild conditions, they have no selectivity for tumor cells and normal cells, which limits the application of enediynes in biological anti-tumor.

人体内正常组织的细胞环境在pH=7.4左右,发生癌变后,其特殊性导致细胞组织所处环境明显偏酸性,有的细胞环境甚至可以达到pH=4.5-5.5。利用肿瘤细胞与正常细胞的的这一不同,可以为合成具有较好细胞选择性的烯二炔药物提供有利条件。 The cell environment of normal tissues in the human body is around pH=7.4. After the occurrence of cancer, its particularity makes the environment of cells and tissues obviously acidic, and some cell environments can even reach pH=4.5-5.5. Utilizing this difference between tumor cells and normal cells can provide favorable conditions for the synthesis of enediyne drugs with better cell selectivity.

发明内容 Contents of the invention

本发明的目的是合成一种新型的在酸性条件下发生反应从而具有抗肿瘤作用的烯二炔化合物。利用肿瘤细胞的酸性环境来诱发烯二炔化合物的抗肿瘤活性。 The purpose of the present invention is to synthesize a novel enediyne compound which reacts under acidic conditions and thus has antitumor effect. The acidic environment of tumor cells is used to induce the antitumor activity of enediynes.

本发明的具体技术方案是:一种烯二炔化合物。 The specific technical scheme of the present invention is: an enediyne compound.

   

其中,R代表官能基团; Wherein, R represents a functional group;

R=R、CR1R2R3、(CR1R2)nX或NR1R2;其中R,R1,R2,R3为氢、卤素、C1-C24烷基链、芳香基团、杂环基团;n为1~6;X为氨基、醚基、酰胺基、巯基、膦基、卤素或酯基。 R=R, CR 1 R 2 R 3 , (CR 1 R 2 ) n X or NR 1 R 2 ; where R, R 1 , R 2 , R 3 are hydrogen, halogen, C1-C24 alkyl chain, aryl group, heterocyclic group; n is 1~6; X is amino, ether, amido, mercapto, phosphino, halogen or ester.

所述芳香基团是苯基、萘基、芘基、蒽基、烷基或卤素;所述杂环基团是吡咯、噻吩、吡啶、哌嗪、吡喃、呋喃、咪唑、吡唑或吗啡啉。 The aromatic group is phenyl, naphthyl, pyrenyl, anthracenyl, alkyl or halogen; the heterocyclic group is pyrrole, thiophene, pyridine, piperazine, pyran, furan, imidazole, pyrazole or morphine phylloline.

上述结构式以马来酰亚胺为主骨架结构。 Above-mentioned structural formula is main skeleton structure with maleimide.

上述结构式含有可以在酸性条件下水解的末端3,3,3 - 三乙氧基官能团。 The above formula contains a terminal 3,3,3-triethoxy functional group that can be hydrolyzed under acidic conditions.

 本发明又一目的是在酸性条件下通过水解反应生成易发生Bergman Cyclization反应的有生物活性的另一种烯二炔: Another object of the present invention is to generate another biologically active enediyne that is prone to Bergman Cyclization reaction by hydrolysis reaction under acidic conditions:

   

本发明提出的烯二炔化合物可作为抗癌药物的应用。所述烯二炔是基于1-苄基-3,4-二(3,3,3-三乙氧基-1-丙炔基)-1H-吡咯-2,5-二酮的化合物。该化合物可以通过结构调控衍生合成出具有不同新功能的同系化合物。研究表明,此类化合物可以在酸性条件下水解生成高活性的烯二炔,并通过自发的Bergman Cyclization反应,造成DNA 的链裂解和肿瘤细胞的死亡。这种新的烯二炔化合物有望在肿瘤抑制以及癌症治疗中发挥潜在作用。 The enediyne compound proposed by the invention can be used as an anticancer drug. The enediyne is a compound based on 1-benzyl-3,4-bis(3,3,3-triethoxy-1-propynyl)-1H-pyrrole-2,5-dione. The compound can be derivatized and synthesized into homologous compounds with different new functions through structure regulation. Studies have shown that such compounds can be hydrolyzed under acidic conditions to generate highly active enediynes, and through the spontaneous Bergman Cyclization reaction, resulting in DNA chain cleavage and tumor cell death. This new enediyne compound is expected to play a potential role in tumor suppression and cancer therapy.

本发明还提供一种烯二炔化合物的制备方法,使用醋酸钯和碳酸铯为主的催化剂,甲苯和四氢呋喃的混合溶剂进行的薗头偶合反应(Sonogashira反应);步骤是: The present invention also provides a kind of preparation method of enediyne compound, uses palladium acetate and cesium carbonate as main catalyst, the sonogashira coupling reaction (Sonogashira reaction) that the mixed solvent of toluene and tetrahydrofuran carries out; The steps are:

在氮气气氛下,将碘化亚铜,醋酸钯,三苯基膦,碳酸铯和3,4 -二碘-N-苄基马来酰亚胺依次加入瓶中,加入除水重蒸过的四氢呋喃和甲苯后搅拌,加入3,3,3 - 三乙氧基-1-丙炔,在45℃下搅拌反应5h后完成反应;将反应后的混合体系直接用硅镁型吸附剂做载体,环己烷/乙酸乙酯作淋洗剂直接进行柱层析分离,通过抽真空除去溶剂后,得到浅黄色粘稠产物烯二炔a,结构如下: Under a nitrogen atmosphere, add cuprous iodide, palladium acetate, triphenylphosphine, cesium carbonate, and 3,4-diiodo-N-benzylmaleimide to the bottle in sequence, and add distilled After THF and toluene were stirred, 3,3,3-triethoxy-1-propyne was added, and the reaction was completed after stirring at 45°C for 5 hours; the mixed system after the reaction was directly used as a carrier of silicon-magnesium-type adsorbent, Cyclohexane/ethyl acetate was used as the eluent to directly carry out column chromatography separation, and after the solvent was removed by vacuuming, the light yellow viscous product enediyne a was obtained, and the structure was as follows:

.

将合成的烯二炔a进行水解反应生成烯二炔b;步骤是: The synthesized enediyne a is hydrolyzed to generate enediyne b; the steps are:

在0 ℃下,将新鲜制备的烯二炔a溶解在三氯甲烷中,加入两滴三氟乙酸后迅速搅拌30秒,然后加入研磨过的碳酸钾粉末以中和反应中过量的三氟乙酸;过滤除掉固体,0 ℃下真空除掉溶剂,得到黄色粘稠固体烯二炔b,结构为: Dissolve freshly prepared enediyne a in chloroform at 0 °C, add two drops of trifluoroacetic acid and stir rapidly for 30 seconds, then add ground potassium carbonate powder to neutralize excess trifluoroacetic acid in the reaction ; The solid was removed by filtration, and the solvent was removed in vacuo at 0 °C to obtain a yellow viscous solid enediyne b with the structure:

.

本发明提供的烯二炔的抗肿瘤生物活性实验,包括: The antitumor biological activity experiment of enediyne provided by the present invention includes:

烯二炔导致超螺旋双链DNA的链断裂实验,主要使用超螺旋质粒ΦX174 RF1DNA来评价烯二炔a和烯二炔b对DNA的活性。将两种不同的烯二炔分别溶于DMSO和丙酮中,加到DNA的TE缓冲液中,在37℃下培养一段时间后进行琼脂糖凝胶电泳,来分析DNA的裂解情况。 Enediyne causes supercoiled double-stranded DNA strand break experiments, mainly using the supercoiled plasmid ΦX174 RF1DNA to evaluate the activity of enediyne a and enediyne b on DNA. Two different enediynes were dissolved in DMSO and acetone respectively, added to the TE buffer of DNA, incubated at 37°C for a period of time, and then subjected to agarose gel electrophoresis to analyze the cleavage of DNA.

烯二炔导致肿瘤细胞凋亡的毒性表明。使用肝癌肿瘤细胞HepG2细胞来评价烯二炔a的抗肿瘤作用。将烯二炔a溶于丙酮中加入到培养好的肿瘤细胞内,培养一段时间后用MTT方法对细胞活性进行评价。 The toxicity of enediynes to induce tumor cell apoptosis has been shown. The antitumor effect of enediyne a was evaluated using HepG2 cells, a hepatocellular carcinoma tumor cell. Dissolve enediyne a in acetone and add it into the cultured tumor cells. After culturing for a period of time, use the MTT method to evaluate the cell viability.

本发明效果,经实验结果发现,烯二炔a在酸性条件下可以导致DNA明显的链裂解;水解后的活性烯二炔b在中性条件下也可以导致DNA的明显链裂解。同时,烯二炔a对肿瘤细胞也表现出很明显的细胞凋亡作用。 The effect of the present invention is that it is found through experiments that enediyne a can cause obvious strand cleavage of DNA under acidic conditions; active enediyne b after hydrolysis can also cause obvious strand cleavage of DNA under neutral conditions. At the same time, enediyne a also showed obvious apoptosis effect on tumor cells.

本发明的优点在于,合成出在酸性条件下能发生Bergman Cyclization的烯二炔。利用人体内肿瘤细胞的酸性环境,这类烯二炔可以导致DNA明显的链断裂以及造成肿瘤细胞的凋亡,说明合成的烯二炔具有较高的生物活性。 The invention has the advantage of synthesizing an enediyne capable of Bergman Cyclization under acidic conditions. Taking advantage of the acidic environment of tumor cells in the human body, this kind of enediyne can cause DNA strand breaks and apoptosis of tumor cells, indicating that the synthesized enediynes have high biological activity.

附图说明 Description of drawings

图1—烯二炔a引起DNA的链断裂。 Figure 1 - Enediyne a causes strand breaks in DNA.

其中:—1—DNA+烯二炔a的DMSO溶液+ PPTS,pH=6 Among them: -1-DMSO solution of DNA + enediyne a + PPTS, pH=6

—2—DNA+ DMSO+ PPTS,pH=6 —2—DNA+ DMSO+ PPTS, pH=6

—3—DNA+烯二炔a的DMSO溶液+ PPTS,pH=5 —3—DMSO solution of DNA + enediyne a + PPTS, pH=5

—4—DNA+ DMSO+ PPTS,pH=5 —4—DNA+ DMSO+ PPTS, pH=5

—5—DNA+烯二炔a的DMSO溶液+ PPTS,pH=4 —5—DMSO solution of DNA+endiyne a+PPTS, pH=4

—6—DNA+ DMSO+ PPTS,pH=4。 —6—DNA+DMSO+PPTS, pH=4.

图2—烯二炔b引起DNA的链断裂。 Figure 2—Enediyne b causes strand breaks in DNA.

其中:—1—DNA Among them:—1—DNA

—2—DNA+ 丙酮 —2—DNA+ acetone

—3—DNA+烯二炔b ,100 mM/L —3—DNA+enediyne b, 100 mM/L

—4—DNA+烯二炔a ,100 mM/L —4—DNA+enediyne a, 100 mM/L

—5—DNA+烯二炔b ,50 mM/L —5—DNA+enediyne b, 50 mM/L

—6—DNA+烯二炔a ,50 mM/L。 —6—DNA+enediyne a, 50 mM/L.

图3—烯二炔a引起HepG2肿瘤细胞的凋亡作用。 Figure 3—Endiyne a induces apoptosis in HepG2 tumor cells.

其中:—□—HepG2+烯二炔a的DMSO溶液(浓度分别为0, 0.15 mM/L, 0.3 mM/L, 0.6 mM/L, 1.3 mM/L, 2.5 mM/L, 5 mM/L, 10 mM/L, 20 mM/L and 40 mM/L)反应时间24小时 Among them: — — DMSO solution of HepG2 + enediyne a (concentrations are 0, 0.15 mM/L, 0.3 mM/L, 0.6 mM/L, 1.3 mM/L, 2.5 mM/L, 5 mM/L, 10 mM/L, 20 mM/L and 40 mM/L) reaction time 24 hours

—○—HepG2+烯二炔a的DMSO溶液(浓度分别为0, 0.15 mM/L, 0.3 mM/L, 0.6 mM/L, 1.3 mM/L, 2.5 mM/L, 5 mM/L, 10 mM/L, 20 mM/L and 40 mM/L)反应时间48小时 — ○—HepG2+ enediyne a DMSO solution (concentrations are 0, 0.15 mM/L, 0.3 mM/L, 0.6 mM/L, 1.3 mM/L, 2.5 mM/L, 5 mM/L, 10 mM/ L, 20 mM/L and 40 mM/L) reaction time 48 hours

—▽—HepG2+烯二炔a的DMSO溶液(浓度分别为0, 0.15 mM/L, 0.3 mM/L, 0.6 mM/L, 1.3 mM/L, 2.5 mM/L, 5 mM/L, 10 mM/L, 20 mM/L and 40 mM/L)反应时间72小时。 —▽—HepG2+ enediyne a DMSO solution (concentrations are 0, 0.15 mM/L, 0.3 mM/L, 0.6 mM/L, 1.3 mM/L, 2.5 mM/L, 5 mM/L, 10 mM/ L, 20 mM/L and 40 mM/L) The reaction time is 72 hours.

具体实施方式 Detailed ways

<例1>烯二炔a的合成 <Example 1> synthesis of enediyne a

合成烯二炔a所用的两种原料3,4 -二碘-N-苄基马来酰亚胺和3,3,3 - 三乙氧基-1 - 丙炔分别根据文献Tetrahedron 2005, 61, 4585和Organic letters 2008, 10, 4915所述方法合成。 The two raw materials 3,4-diiodo-N-benzylmaleimide and 3,3,3-triethoxy-1-propyne used in the synthesis of enediyne a are respectively according to the literature Tetrahedron 2005 , 61, 4585 and Organic letters 2008 , 10, 4915 described method synthesis.

在氮气下,将碘化亚铜(98.5mg,40mmol%) ,醋酸钯(29.2mg,10mmol%), 三苯基膦(68.2mg,20mmol%),碳酸铯(1.27g,3.9mmol)和3,4 -二碘-N-苄基马来酰亚胺(0.57g,1.3mmol)依次加到干燥过的Schlenk瓶中,加入重蒸除水的四氢呋喃(4ml)和甲苯(10ml)后搅拌,加入3,3,3 - 三乙氧基-1-丙炔(0.68g, 3.9mmol),在45℃下搅拌反应5h后完成反应。将反应体系粗溶液用环己烷/乙酸乙酯=40:1的淋洗剂进行硅镁型吸附剂的柱层析分离,通过真空除去溶剂后,得到浅黄色粘稠产物烯二炔a(0.405g, 产率59.6%)。1H NMR( 500MHz, CDCl3):δ(ppm) 7.28-7.34 (m, 5H, Bn), 4.69 (s, 2H, CH2), 3.73 (q,12H, CH2, J=7.0MHz), 1.25(t, 18H,CH3, J=7.0MHz)。 13C NMR( 500MHz, CDCl3, ppm): δ165.6, 135.3, 128.9, 128.8, 128.6, 128.2, 109.1, 103.7, 72.9, 59.5, 42.7, 14.9。HRMS ( ESI ): m/z C29H37NO8( M+Na) + ,计算值550.2417; 实测值 550.2419。结构式为: Under nitrogen, copper iodide (98.5mg, 40mmol%), palladium acetate (29.2mg, 10mmol%), triphenylphosphine (68.2mg, 20mmol%), cesium carbonate (1.27g, 3.9mmol%) and 3 , 4-diiodo-N-benzylmaleimide (0.57g, 1.3mmol) was successively added to a dried Schlenk bottle, added tetrahydrofuran (4ml) and toluene (10ml) which had been distilled to remove water, and stirred, 3,3,3-Triethoxy-1-propyne (0.68g, 3.9mmol) was added, and the reaction was completed after stirring at 45°C for 5h. The crude solution of the reaction system was separated by column chromatography on a silicon-magnesium-type adsorbent with a eluent of cyclohexane/ethyl acetate=40:1, and after the solvent was removed by vacuum, the light yellow viscous product enediyne a ( 0.405g, yield 59.6%). 1 H NMR( 500MHz, CDCl 3 ):δ(ppm) 7.28-7.34 (m, 5H, Bn), 4.69 (s, 2H, CH 2 ), 3.73 (q,12H, CH 2, J=7.0MHz), 1.25(t, 18H, CH 3 , J=7.0MHz). 13 C NMR (500MHz, CDCl 3 , ppm): δ165.6, 135.3, 128.9, 128.8, 128.6, 128.2, 109.1, 103.7, 72.9, 59.5, 42.7, 14.9. HRMS (ESI): m/z calcd for C29H37NO8 (M+Na) + , 550.2417 ; found 550.2419 . The structural formula is:

<例2>烯二炔b的合成 <Example 2> synthesis of enediyne b

反应在0 ℃进行。 将新鲜制备的烯二炔a(20mg)溶解在三氯甲烷(10ml )中,加入两滴三氟乙酸(TFA)。快速搅拌30秒,然后加入研磨过的碳酸钾粉末(2g) ,以中和反应中过量的酸。过滤除去固体,收集滤液,真空除掉溶剂,得到黄色的烯二炔b。1H NMR( 500MHz, CDCl3): δ(ppm)7.30-7.35(m,5H,Bn), 4.72(s,2H,CH2),4.33(q,4H,CH2,J=7.0MHz),1.36(t,6H,CH3,J=7.0MHz).13C NMR ( 500MHz, CDCl3): δ(ppm)164.4, 152.0, 134.9, 129.4, 128.9, 128.7, 128.4, 98.5, 72.6, 63.1, 43.0, 13.9。HRMS ( ESI ): m/z C21H17NO6( M+Na) +,计算值402.0954; 实测值 402.0958。结构式为: The reaction was carried out at 0 °C. Freshly prepared enediyne a (20 mg) was dissolved in chloroform (10 ml) and two drops of trifluoroacetic acid (TFA) were added. After stirring rapidly for 30 seconds, ground potassium carbonate powder (2 g) was added to neutralize excess acid in the reaction. The solid was removed by filtration, the filtrate was collected, and the solvent was removed in vacuo to give enediyne b as yellow. 1 H NMR( 500MHz, CDCl 3 ): δ(ppm) 7.30-7.35(m,5H,Bn), 4.72(s,2H,CH 2 ),4.33(q,4H,CH 2, J=7.0MHz), 1.36(t,6H,CH 3, J=7.0MHz). 13 C NMR ( 500MHz, CDCl 3 ): δ(ppm) 164.4, 152.0, 134.9, 129.4, 128.9, 128.7, 128.4, 98.5, 72.6, 63.1, 43.0 , 13.9. HRMS (ESI): m/z calcd for C21H17NO6 (M+Na) + , 402.0954 ; found 402.0958. The structural formula is:

<例3>烯二炔a对超螺旋双链DNA的裂解能力 <Example 3> Cleavage ability of enediyne a to supercoiled double-stranded DNA

选用超螺旋质粒ΦX174 RF1DNA进行实验。将烯二炔a的DMSO溶液加到ΦX174的TE缓冲液中,加入对甲苯磺酸吡啶盐(PPTS)的水溶液调节体系pH分别在pH=4,pH=5, pH=6左右。将混合物放在37 ℃下恒温40小时后,取出溶液进行琼脂糖凝胶电泳,通过紫外成像系统进行扫描成像分析。 The supercoiled plasmid ΦX174 RF1DNA was used for the experiment. Add the DMSO solution of enediyne a to the TE buffer solution of ΦX174, and add the aqueous solution of pyridinium p-toluenesulfonate (PPTS) to adjust the pH of the system at pH=4, pH=5, and pH=6 respectively. After the mixture was kept at 37°C for 40 hours, the solution was taken out for agarose gel electrophoresis, and scanned and imaged by an ultraviolet imaging system.

ΦX174 RF1DNA是一种超螺旋双链DNA,其95%以FormⅠ形式存在,如果DNA被裂解,会由FormⅠ转化为FormⅡ,称为单链裂解,或者由FormⅠ转化成FormⅢ,称为双链裂解。 ΦX174 RF1DNA is a supercoiled double-stranded DNA, 95% of which exists in the form of Form I. If the DNA is cleaved, it will be converted from Form I to Form II, which is called single-strand cleavage, or from Form I to Form III, which is called double-strand cleavage.

结果表明,在pH=6时,DNA无明显的变化,在pH=5时DNA开始有所变化,并且加入烯二炔a的DNA比不加烯二炔a的DNA变化略明显。在 pH=4时,加入烯二炔a后的DNA发生明显的变化,表明烯二炔a造成了DNA的单链裂解,DNA由FormⅠ形式转化为FormⅡ形式。如图1所示。 The results showed that DNA did not change significantly at pH=6, and DNA began to change at pH=5, and the DNA with added enediyne a changed slightly more than the DNA without added enediyne a. At pH = 4, the DNA after adding enediyne a changed significantly, indicating that enediyne a caused DNA single-strand cleavage, and the DNA was converted from Form I to Form II. As shown in Figure 1.

<例4>烯二炔b对超螺旋双链DNA的裂解能力 <Example 4> cleavage ability of enediyne b to supercoiled double-stranded DNA

将烯二炔b的丙酮溶液加到ΦX174的TE缓冲液中,将混合物放在37 ℃下恒温反应48小时后,取溶液进行琼脂糖凝胶电泳,通过紫外成像系统进行扫描成像分析。 The acetone solution of enediyne b was added to the TE buffer of ΦX174, and the mixture was kept at 37 °C for 48 hours to react at a constant temperature. The solution was taken for agarose gel electrophoresis, and scanned and imaged by an ultraviolet imaging system.

结果表明,当烯二炔a被水解成活性更高的烯二炔b后,在中性条件下,浓度为100mM/L的烯二炔b就可以导致DNA的明显裂解,即使浓度降低到50mM/L,DNA也有明显的链断裂。如图2所示。 The results show that when enediyne a is hydrolyzed into more active enediyne b, under neutral conditions, enediyne b at a concentration of 100 mM/L can cause significant cleavage of DNA, even if the concentration is reduced to 50 mM /L, the DNA also had obvious strand breaks. as shown in picture 2.

<例5>烯二炔a诱导肿瘤细胞的凋亡作用 <Example 5> Apoptosis of tumor cells induced by enediyne a

使用MTT分析方法对细胞活性进行表征,具体方法为: The cell viability was characterized by MTT analysis method, the specific method is as follows:

1)  在37 ℃含5%CO2的气氛下,在补充有10 %胎牛血清的DMEM培养基中培养细胞24h; 1) Cultivate cells in DMEM medium supplemented with 10% fetal bovine serum at 37°C in an atmosphere containing 5% CO 2 for 24 hours;

2)  用含0.25%胰蛋白酶和0.03% EDTA的缓冲盐溶液将细胞胰蛋白酶化处理,直至达到70 %汇合在组织培养瓶中; 2) Trypsinize the cells with a buffered saline solution containing 0.25% trypsin and 0.03% EDTA until they reach 70% confluency in tissue culture flasks;

3)  将细胞接种在96孔细胞培养板上,并使其贴壁培养24小时; 3) Seed the cells on a 96-well cell culture plate and allow them to adhere to the wall for 24 hours;

4)  用药物处理细胞,将烯二炔溶在丙酮中加到细胞培养基中;继续培养24h,48h,72h; 4) Treat cells with drugs, dissolve enediyne in acetone and add to cell culture medium; continue to culture for 24h, 48h, 72h;

5)  将溶有MTT(3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐)的PBS溶液(pH=7.4)加入到每个孔板中,将细胞再培养4小时,使黄色染料变成蓝色晶体; 5) Add the PBS solution (pH=7.4) dissolved with MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide) into each well plate , incubate the cells for another 4 hours to turn the yellow dye into blue crystals;

6)  通过抽吸除去未反应的染料,在孔板中加入DMSO进行晶解; 6) Remove the unreacted dye by suction, and add DMSO to the well plate for crystallization;

7)  细胞活性检测:用酶联免疫检测仪在490nm波长处测定OD值,间接反映细胞的活性。 7) Detection of cell activity: Measure the OD value at a wavelength of 490nm with an enzyme-linked immunosorbent assay, which indirectly reflects the activity of the cells.

结果表明,用烯二炔a处理肿瘤细胞后,培养24h后即可导致肿瘤细胞明显的凋亡,培养时间到48h,72h后,细胞活性有明显的降低,如图3所示。 The results showed that treatment of tumor cells with enediyne a could lead to obvious apoptosis of tumor cells after 24 hours of culture, and after 48 hours and 72 hours of culture, the cell viability was significantly reduced, as shown in Figure 3 .

Claims (10)

1.一种烯二炔化合物,其化学结构式是: 1. An enediyne compound, its chemical structural formula is: 其中,R代表官能基团; Wherein, R represents a functional group; R=R、CR1R2R3、(CR1R2)nX或NR1R2;其中R,R1,R2,R3为氢、卤素、C1-C24烷基链、芳香基团、杂环基团;n为1~6;X为氨基、醚基、酰胺基、巯基、膦基、卤素或酯基。 R=R, CR 1 R 2 R 3 , (CR 1 R 2 ) n X or NR 1 R 2 ; where R, R 1 , R 2 , R 3 are hydrogen, halogen, C1-C24 alkyl chain, aryl group, heterocyclic group; n is 1~6; X is amino, ether, amido, mercapto, phosphino, halogen or ester. 2.根据权利要求1所述的烯二炔化合物,其特征在于,所述芳香基团是苯基、萘基、芘基、蒽基、烷基或卤素;所述杂环基团是吡咯、噻吩、吡啶、哌嗪、吡喃、呋喃、咪唑、吡唑或吗啡啉。 2. enediyne compound according to claim 1, is characterized in that, described aromatic group is phenyl, naphthyl, pyrenyl, anthracenyl, alkyl or halogen; Described heterocyclic group is pyrrole, Thiophene, pyridine, piperazine, pyran, furan, imidazole, pyrazole or morpholine. 3.根据权利要求1或2所述的烯二炔化合物,其特征是以马来酰亚胺为主骨架结构。 3. The enediyne compound according to claim 1 or 2, characterized in that maleimide is the main skeleton structure. 4.根据权利要求1货2所述的烯二炔化合物,其特征是含有可以在酸性条件下水解的末端3,3,3 - 三乙氧基官能团。 4. The enediyne compound according to claim 1 or 2, characterized in that it contains a terminal 3,3,3-triethoxy functional group that can be hydrolyzed under acidic conditions. 5.一种烯二炔化合物的制备方法,其反应如下: 5. a preparation method of enediyne compound, its reaction is as follows: . 6.根据权利要求1所述的烯二炔化合物,其结构式是: 6. enediyne compound according to claim 1, its structural formula is: . 7.一种烯二炔化合物的制备方法,使用醋酸钯和碳酸铯为主的催化剂,甲苯和四氢呋喃的混合溶剂进行的薗头偶合反应;步骤是: 7. a kind of preparation method of enediyne compound, the catalyzer that uses palladium acetate and cesium carbonate as main catalyzer, the mixed solvent of toluene and tetrahydrofuran carries out the sorrel coupling reaction; Step is: 在氮气气氛下,将碘化亚铜,醋酸钯,三苯基膦,碳酸铯和3,4 -二碘-N-苄基马来酰亚胺依次加入瓶中,加入除水重蒸过的四氢呋喃和甲苯后搅拌,加入3,3,3 - 三乙氧基-1-丙炔,在45℃下搅拌反应5h后完成反应;将反应后的混合体系直接用硅镁型吸附剂做载体,环己烷/乙酸乙酯作淋洗剂直接进行柱层析分离,通过抽真空除去溶剂后,得到浅黄色粘稠产物烯二炔a,结构如下: Under a nitrogen atmosphere, add cuprous iodide, palladium acetate, triphenylphosphine, cesium carbonate, and 3,4-diiodo-N-benzylmaleimide to the bottle in sequence, and add distilled After THF and toluene were stirred, 3,3,3-triethoxy-1-propyne was added, and the reaction was completed after stirring at 45°C for 5 hours; the mixed system after the reaction was directly used as a carrier of silicon-magnesium-type adsorbent, Cyclohexane/ethyl acetate was used as the eluent to directly carry out column chromatography separation, and after the solvent was removed by vacuuming, the light yellow viscous product enediyne a was obtained, and the structure was as follows: . 8.根据权利要求5所述的烯二炔的制备方法,其特征是将合成的烯二炔a进行水解反应生成烯二炔b;步骤是: 8. the preparation method of enediyne according to claim 5 is characterized in that synthetic enediyne a is hydrolyzed to generate enediyne b; the steps are: 在0 ℃下,将新鲜制备的烯二炔a溶解在三氯甲烷中,加入两滴三氟乙酸后迅速搅拌30秒,然后加入研磨过的碳酸钾粉末以中和反应中过量的三氟乙酸;过滤除掉固体,0 ℃下真空除掉溶剂,得到黄色粘稠固体烯二炔b,结构为: Dissolve freshly prepared enediyne a in chloroform at 0 °C, add two drops of trifluoroacetic acid and stir rapidly for 30 seconds, then add ground potassium carbonate powder to neutralize excess trifluoroacetic acid in the reaction ; The solid was removed by filtration, and the solvent was removed in vacuo at 0 °C to obtain a yellow viscous solid enediyne b with the structure: . 9.一种如权利要求1所述的烯二炔用于抗肿瘤药物中的应用。 9. The application of an enediyne as claimed in claim 1 in antitumor drugs. 10.一种如权利要求6所述的烯二炔(a)用于抗肿瘤药物中的应用。 10. The use of an enediyne (a) as claimed in claim 6 in antitumor drugs.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106632411A (en) * 2016-12-23 2017-05-10 浙江大学常州工业技术研究院 Antitumor fluorescent compound, and preparation method and applications thereof

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050004212A1 (en) * 2003-06-30 2005-01-06 Kaohsiung Medical University Aryl-substituted acyclic enediyne compounds
US20050004211A1 (en) * 2003-06-30 2005-01-06 Kaohsiung Medical University Pharmaceutical compositions comprising aryl-substituted acyclic enediyne compounds
US6987132B1 (en) * 2000-09-25 2006-01-17 Advanced Research And Technology Institute, Inc. Enediyne compounds and methods related thereto
CN101163672A (en) * 2005-03-23 2008-04-16 国立大学法人东京工业大学 Monomolecular conductive complex, conductive self-assembled film, and metal-semiconductor-containing electrode assembly using same
CN102515141A (en) * 2011-12-02 2012-06-27 东华理工大学 Method for synthesizing modified SWCNTs (Single Wall Carbon Nano Tubes) based on Bergman cyclizing reaction

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6987132B1 (en) * 2000-09-25 2006-01-17 Advanced Research And Technology Institute, Inc. Enediyne compounds and methods related thereto
US20050004212A1 (en) * 2003-06-30 2005-01-06 Kaohsiung Medical University Aryl-substituted acyclic enediyne compounds
US20050004211A1 (en) * 2003-06-30 2005-01-06 Kaohsiung Medical University Pharmaceutical compositions comprising aryl-substituted acyclic enediyne compounds
CN101163672A (en) * 2005-03-23 2008-04-16 国立大学法人东京工业大学 Monomolecular conductive complex, conductive self-assembled film, and metal-semiconductor-containing electrode assembly using same
CN102515141A (en) * 2011-12-02 2012-06-27 东华理工大学 Method for synthesizing modified SWCNTs (Single Wall Carbon Nano Tubes) based on Bergman cyclizing reaction

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
AGATHE SOUFFRIN,等: "Efficient and General Protocol for Sonogashira Cross-Coupling Reactions of Maleimides", 《EUROPEAN JOURNAL OF ORGANIC CHEMISTRY》, no. 13, 31 December 2012 (2012-12-31), pages 2499 - 2502 *
FABIO BELLINA,等: "Regioselective synthesis of cytotoxic 4-(1-alkynyl)-substituted 2-(5H)-furanones", 《TETRAHEDRON》, vol. 59, no. 46, 31 December 2003 (2003-12-31), pages 9091 - 9100, XP004470349, DOI: doi:10.1016/j.tet.2003.09.061 *
刘文: "烯二炔类抗肿瘤抗生素的生物合成研究进展", 《世界科技研究与发展》, vol. 27, no. 3, 30 June 2005 (2005-06-30), pages 24 - 31 *
李菲,等: "通过Bergman环化聚合反应合成侧链含有苯丙氨酸的手性聚合物", 《华东理工大学学报(自然科学版)》, vol. 39, no. 2, 30 April 2013 (2013-04-30), pages 121 - 125 *
邵荣光,等: "烯二炔类抗肿瘤药物的研究与开发", 《中国处方药》, no. 12, 31 December 2006 (2006-12-31), pages 6 - 9 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106632411A (en) * 2016-12-23 2017-05-10 浙江大学常州工业技术研究院 Antitumor fluorescent compound, and preparation method and applications thereof
CN106632411B (en) * 2016-12-23 2018-09-04 浙江大学常州工业技术研究院 A kind of antitumor fluorescent chemicals, preparation method and the usage

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