WO2017128981A1 - 一种具有抗肿瘤作用的化合物及其制备方法和应用 - Google Patents

一种具有抗肿瘤作用的化合物及其制备方法和应用 Download PDF

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WO2017128981A1
WO2017128981A1 PCT/CN2017/071276 CN2017071276W WO2017128981A1 WO 2017128981 A1 WO2017128981 A1 WO 2017128981A1 CN 2017071276 W CN2017071276 W CN 2017071276W WO 2017128981 A1 WO2017128981 A1 WO 2017128981A1
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compound
cancer
pharmaceutically acceptable
preparation
tba
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French (fr)
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雷海民
王鹏龙
徐冰
熊磊
龚兆龙
林毅晖
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四川思路迪药业有限公司
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Priority to CN201780003139.3A priority Critical patent/CN108137644B/zh
Publication of WO2017128981A1 publication Critical patent/WO2017128981A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
    • C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0087—Galenical forms not covered by A61K9/02 - A61K9/7023
    • A61K9/0095—Drinks; Beverages; Syrups; Compositions for reconstitution thereof, e.g. powders or tablets to be dispersed in a glass of water; Veterinary drenches
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
    • A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/28—Dragees; Coated pills or tablets, e.g. with film or compression coating
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms

Definitions

  • the invention relates to a compound and a preparation method and application thereof, in particular to a compound having antitumor activity and a preparation method and application thereof.
  • cancer death especially the multi-type malignant cancers such as liver cancer and colon cancer.
  • chemotherapy is one of the main ways of cancer, and first-line chemotherapy drugs are Killing cancer cells often causes serious toxic side effects on normal organisms such as nephrotoxicity, hepatotoxicity, neurotoxicity and the like. It is the goal of pharmacy workers to obtain anti-cancer drugs with high efficiency, low toxicity and strong selectivity. It is gratifying that the latest researches at home and abroad have found that compounds with strong cytotoxicity have good drug-forming properties, which is in line with the development trend of contemporary precision medicine. (Cancer Cell, 2015, 28, 240-252; Int. J. Mol. Sci. 2015, 16(7), 16401-16413).
  • the invention uses the betulinic acid (BA) having antitumor and anti-hepatic disease biological activity and the natural drug ligustrazine (TMP) as a starting material to chemically combine and synthesize the compound of the present invention.
  • BA betulinic acid
  • TMP natural drug ligustrazine
  • the activity evaluation of this kind of compound mainly focused on anti-tumor (especially liver cancer, intestinal cancer, etc.), and tested the analogs against five cancer cell lines (HepG-2, HT-29, Hela, BGC-823, A549). And cytotoxic activity of non-cancer cell lines (human liver normal cells L02).
  • a first object of the present invention is to provide a compound having the structure of Formula 1 and a process for the preparation thereof.
  • a second object of the invention is to provide the use of a compound of formula 1 for the preparation of an anti-tumor drug.
  • a third object of the present invention is to provide a pharmaceutical composition having an antitumor effect.
  • the compound may be added to a conventional excipient in the field of preparation to prepare a conventional dosage form such as a tablet, a capsule, a granule, a powder, an oral solution, an injection or the like.
  • the preparation method of the compound of the invention comprises the following steps:
  • Step 1 dissolving betulinic acid in an organic solvent, and 2-chloromethyl-3,5,6-trimethylpyrazine to form an intermediate compound ligustrazine-white betearate under basic conditions (middle Compound TBA);
  • Step 2 the intermediate compound TBA reacts with sarcosine under the action of a catalyst to form a compound LQC-C.
  • the above reaction is carried out at -20 ° C to 250 ° C;
  • the organic solvent is an ether, an alcohol, an alkane, an aromatic hydrocarbon, a ketone, an alkyl halide, an amide, a nitrile, an ester having 1 to 20 carbon atoms or various ratios thereof a mixture;
  • the base used is triethylamine or potassium carbonate;
  • the catalyst is 1-hydroxybenzotriazole (HOBT), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride Salt (EDCI), 1,3-dicyclohexylcarbodiimide (DCC) or 4-dimethylaminopyridine (DMAP).
  • HOBT 1-hydroxybenzotriazole
  • EDCI 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride Salt
  • DCC 1,3-dicyclohexylcarbodiimide
  • the molar ratio of the raw material to the chloropipestazine is 1:0.1 to 1:10; the molar ratio of the raw material to the base is 1:0.1 to 1:10; the molar ratio of the intermediate to the sarcosine is 1:0.1 to 1:10; the molar ratio of the intermediate to the catalyst is from 1:0.1 to 1:10.
  • the reaction route of the present invention is:
  • the invention also provides the use of a compound of formula 1 or a pharmaceutically acceptable salt thereof for the preparation of an anti-tumor drug.
  • the tumor is a liver cancer, a lung cancer, a colon cancer, a cervical cancer, and a gastric cancer cell line.
  • the invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising a formula present in a therapeutically effective amount A mixture of a compound or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient.
  • composition further comprises at least one conventional anticancer drug.
  • the anticancer drug is selected from the group consisting of cyclophosphamide, 5-fluorouracil, paclitaxel, doxorubicin, etoposide, irinotecan, oxaliplatin, cisplatin or Gemcitabine.
  • the invention also provides a method of treating cancer comprising administering to a patient an effective amount of a compound of formula 1 or a pharmaceutically acceptable salt thereof.
  • fillers include: starch, pregelatinized starch, lactose, mannitol, chitin, microcrystalline cellulose, sucrose, etc.
  • disintegrants include: starch, pregelatinized starch, microcrystalline cellulose, carboxymethyl Starch sodium, cross-linked polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, croscarmellose sodium, etc.
  • lubricants include: magnesium stearate, sodium lauryl sulfate, talc, silica, etc.
  • Suspensions include: polyvinylpyrrolidone, microcrystalline cellulose, sucrose, agar, hydroxypropyl methylcellulose, etc.
  • binders include, starch slurry, polyvinylpyrrolidone, hydroxypropyl methylcellulose, etc.
  • sweetness The agent includes: sodium saccharin, aspartame, sucrose, cyclamate, glycyrrhetinic acid, etc.
  • flavoring agents include: sweeteners and various flavors
  • preservatives include: parabens, benzoic acid, sodium benzoate, sorbus Acid and its salts, benzalkonium bromide, chlorine acetate Set, eucalyptus oil and so on.
  • “Pharmaceutically acceptable” as used herein means that the compound or composition must be chemically and/or toxicologically compatible with the other ingredients included in the formulation.
  • the "therapeutically effective amount” means an amount of a compound of the present invention that treats or prevents a particular disease or condition; ameliorating, ameliorating or eliminating one or more symptoms of a particular disease; or preventing or delaying the onset of a particular disease or condition.
  • the compounds of the present invention have activity which significantly inhibits the growth of tumor cell lines (HepG-2, HT-29, Hela, BGC-823, A549) but are less toxic to human liver normal cells (L02).
  • tumor cell lines HepG-2, HT-29, Hela, BGC-823, A549
  • L02 human liver normal cells
  • the anti-proliferative activity of compound LQC-C against human colon cancer cell line HT-29, human cervical cancer cell line Hela and human lung cancer cell line A549 was better than that of positive drug cisplatin; but the cytotoxicity to human liver normal cell L02 was obvious. Lower than the positive drug cisplatin.
  • Experimental drug the compound LQC-C of the present invention (prepared according to Examples 1-4); the raw material drug ligustrazine (TMP), betulinic acid (BA); cisplatin (DPP) for positive drug injection (301001 CF; Qilu Pharmaceutical Co., Ltd.) ).
  • HepG2, HT-29, Hela, BGC-823, A549 and L02 cells were cultured in 1640 medium containing 10% fetal bovine serum, and incubated at 37 ° C in a 5% CO 2 incubator. The cells were all grown in an adherent state, and the growth was observed under an inverted microscope, and subcultured when the number of cells was appropriate.
  • HepG2, HT-29, Hela, BGC-823, A549 and L02 cells in logarithmic growth phase were seeded in 96-well culture plates at a number of 3 ⁇ 103/well in a humidified incubator containing 5% CO 2
  • the cells were cultured at 37 ° C for 24 h; 100 ⁇ L of the test compound was added to each well to give final concentrations of 40.0, 20.0, 10.0, 5.0, 2.5, and 1.25 ⁇ M, respectively.
  • the cell control group and the blank control group were set.
  • the drug group was repeated 4 wells per concentration, and the cell control group and the blank control group were repeated 3 wells.
  • Inhibition rate (%) [1 - (A administration - A blank) / (A normal - A blank)] ⁇ 100%.
  • the IC 50 values of the compound of the present invention LQC-C against five tumor cell lines (HepG2, HT-29, Hela, BGC-823, A549) and human normal liver cells L02 are shown in Table 1.
  • the compound LQC-C showed good inhibition of tumor cell proliferation activity against various cancer cells, and its antitumor activity was superior to that of the raw material; among them, the compound LQC-C on human colon cancer cell line Anti-proliferative activity of HT-29 and human cervical cancer cell line Hela and human lung cancer cell line A549 is better than yang The drug cisplatin; but the cytotoxicity of human liver normal cell L02 is significantly lower than the positive drug cisplatin.
  • the compounds of the invention exhibit an activity of inhibiting the proliferation of tumor cell lines (HepG2, HT-29, Hela, BGC-823, A549).
  • Ligustrazine 21.76 g (160 mmol) (Compound 1) was dissolved in 50 ml of glacial acetic acid, and 18 ml (160 mmol) of 30% hydrogen peroxide was added to react at 90 ° C for 4 h, then 18 ml (160 mmol) of 30% hydrogen peroxide was added to continue the reaction for 2 h, TLC After the reaction was completed, it was cooled to room temperature, adjusted to pH 10 with 50% sodium hydroxide, extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, and solvent was evaporated to afford white liperazine monooxybenzene (compound 2).

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Abstract

提供如式（I）结构的化合物(LQC-C)及其制备方法和在制备抗肿瘤药物中的应用。该化合物具有明显抑制肿瘤细胞系(HepG-2、HT-29、Hela、BGC-823、A549)生长的活性，但对人肝正常细胞(L02)系毒性较小。其中，化合物的对人结肠癌细胞系HT-29、人宫颈癌细胞系Hela以及人肺癌细胞系A549抗增殖活性优于阳性药顺铂；但对人肝正常细胞L02的细胞毒性明显低于阳性药顺铂。

Description

一种具有抗肿瘤作用的化合物及其制备方法和应用 技术领域
本发明涉及一种化合物及其制备方法和应用，具体涉及一种具有抗肿瘤作用性的化合物及其制备方法和应用。
背景技术
当前临床上癌症的治疗往往出现“癌死人亡”的悲剧，尤其是肝癌、结肠癌等多类恶性癌变目前尚未特效药物，究其原因是目前化疗是癌症的主要方式之一，一线化疗药物在杀伤癌细胞的同时往往对正常机体造成严重的毒副作用，例如：肾毒性、肝毒性、神经毒性等等。对于获得高效、低毒、选择性强的抗癌药物是药学工作者孜孜以求的目标，可喜的是国内外最新研究发现细胞毒选择性强的化合物具有良好的成药性，符合当代精准医学的发展趋势(Cancer Cell,2015,28,240–252；Int.J.Mol.Sci.2015,16(7),16401-16413)。
本发明以具有抗肿瘤、抗肝病生物活性的白桦脂酸(BA)及天然药物川芎嗪(TMP)为起始原料，进行化学拼合合成本发明化合物。对该类化合物的活性评价主要围绕抗肿瘤(尤其肝癌、肠癌等)方面展开，分别测试了类似物对5种癌症细胞系(HepG-2、HT-29、Hela、BGC-823、A549)及非癌细胞系(人肝正常细胞L02)的细胞毒活性。
发明内容
本发明的第一个目的在于提供具有式1结构的化合物及其制备方法。
本发明的第二个目的在于提供式1化合物在制备抗肿瘤药物中的应用。
本发明的第三个目的在于提供一种具有抗肿瘤作用的药物组合物。
本发明的目的是通过如下技术方案实现的：
具有式1结构的化合物或其药学上可接受的盐，
Figure PCTCN2017071276-appb-000001
进一步，所述化合物可加入制剂领域常规辅料制成片剂、胶囊剂、颗粒剂、散剂、口服液、注射剂等常规剂型。
本发明所述化合物的制备方法包括如下步骤：
步骤1，将白桦脂酸溶解于有机溶剂中，与2-氯代甲基-3,5,6-三甲基吡嗪在碱性条件下生成中间体化合物川芎嗪-白桦脂酸酯(中间体化合物TBA)；
步骤2，中间体化合物TBA在催化剂的作用下与肌氨酸发生反应生成化合物LQC-C。
进一步，上述反应在-20℃至250℃下进行；所述有机溶剂是含有1-20个碳原子的醚、醇、烷烃、芳香烃、酮、卤代烷、酰胺、腈、酯或者它们各种比例的混合物；所用碱为三乙胺或碳酸钾；所述催化剂为1-羟基苯并三唑(HOBT)、1-乙基-3-(3-二甲胺丙基)碳二亚胺盐酸盐(EDCI)、1,3-二环己基碳二亚胺(DCC)或4-二甲氨基吡啶(DMAP)。
进一步，上述制备方法中，原料与氯代川芎嗪的摩尔比为1:0.1～1:10；原料与碱的摩尔比为1:0.1～1:10；中间体与肌氨酸的摩尔比为1:0.1～1:10；中间体与催化剂的摩尔比为1:0.1～1:10。
本发明反应路线为：
Figure PCTCN2017071276-appb-000002
反应条件和试剂:(a)AcOH,30％H2O2,reflux,90℃,6h；(b)Ac2O,reflux,105℃,2h；(c)THF:MeOH:H2O＝3:1:1,NaOH,1h；(d),THF,Tscl,TEA,DMAP,12h.；(e)DMF,dry K2CO3,60℃,12h.(f)BOC-Sarcosine,EDCI,DMAP,DCM,25℃,12h；(g)HCl(g)in EA,0℃,1h.
本发明还提供式1化合物或其药学上可接受的盐在制备抗肿瘤药物中的应用。
进一步，所述肿瘤为肝癌、肺癌、结肠癌、宫颈癌和胃癌细胞系。
本发明还提供一种药物组合物，该组合物包含以治疗有效量存在的式1化 合物或其药学可接受的盐与至少一种药学可接受的赋形剂的混合物。
进一步，所述组合物还包含至少一种常规抗癌药。
更进一步，所述抗癌药选自环磷酰胺、5-氟尿嘧啶、紫杉醇、阿霉素、依托泊苷、伊立替康、奥沙利铂、顺铂或健择。
本发明还提供治疗癌症的方法，包括给予患者给药有效量的式1化合物或其药学上可接受的盐。
为使上述剂型能够实现，需在制备这些剂型时加入药学可接受的赋形剂，例如填充剂、崩解剂、润滑剂、助悬剂、粘合剂、甜味剂、矫味剂、防腐剂等，填充剂包括：淀粉、预胶化淀粉、乳糖、甘露醇、甲壳素、微晶纤维素、蔗糖等，崩解剂包括：淀粉、预胶化淀粉、微晶纤维素、羧甲基淀粉钠、交联聚乙烯吡咯烷酮、低取代羟丙纤维素、交联羧甲基纤维素钠等，润滑剂包括：硬脂酸镁、十二烷基硫酸钠、滑石粉、二氧化硅等，助悬剂包括：聚乙烯吡咯烷酮、微晶纤维素、蔗糖、琼脂、羟丙基甲基纤维素等，粘合剂包括，淀粉浆、聚乙烯吡咯烷酮、羟丙基甲基纤维素等，甜味剂包括：糖精钠、阿斯帕坦、蔗糖、甜蜜素、甘草次酸等，矫味剂包括：甜味剂及各种香精，防腐剂包括：尼泊金类、苯甲酸、苯甲酸钠、山梨酸及其盐类、苯扎溴铵、醋酸氯乙定、桉叶油等。
本发明所述“药学可接受的”表示化合物或组合物必须在化学上和/或毒理学上与制剂中包含的其他成分相容。
所述“治疗有效量”表示本发明化合物治疗或预防特定疾病或症状；减弱、改善或消除特定疾病的一种或多种症状；或预防或延迟特定疾病或症状的发作的量。
本发明化合物具有明显抑制肿瘤细胞系(HepG-2、HT-29、Hela、BGC-823、A549)生长的活性但对人肝正常细胞(L02)系毒性较小。其中，化合物LQC-C的对人结肠癌细胞系HT-29、人宫颈癌细胞系Hela以及人肺癌细胞系A549抗增殖活性优于阳性药顺铂；但对人肝正常细胞L02的细胞毒性明显低于阳性药顺铂。
实验例1MTT法观察本发明化合物LQC-C对癌细胞及非癌细胞系的增殖影响
1.仪器与材料
Thermo 3111型CO2培养箱；HFsafe生物安全柜；Multiskan GO酶标仪；京立牌LD5-2B型台式低速离心机；Olympus IX71倒置荧光显微镜改良型RPMI-1640培养基、胎牛血清、0.25％胰蛋白酶溶液、噻唑蓝、磷酸盐缓冲液(赛默飞世尔生物化学制品北京有限公司)；Amresco二甲基亚砜(DMSO)；
人肝癌细胞系HepG-2；人结肠癌细胞系HT-29；人宫颈癌细胞系Hela；人胃癌细胞系BGC-823；人肺癌细胞系A549；人肝正常细胞L02；
实验药物：本发明化合物LQC-C(按实施例1-4制备)；原料药川芎嗪(TMP)、白桦脂酸(BA)；阳性药物注射用顺铂(DPP)(301001CF；齐鲁制药有限公司)。
2.方法
2.1不同细胞株的培养
HepG2、HT-29、Hela、BGC-823、A549和L02细胞培养在含10％胎牛血清的1640培养液中，放置于37℃、5％的CO2培养箱中温育。细胞均呈贴壁状态生长，在倒置显微镜下观察生长状况，待细胞数量适量时传代培养。
2.2对肿瘤细和正常细胞的增殖抑制作用
取对数生长期的HepG2、HT-29、Hela、BGC-823、A549和L02细胞，以3×103/孔的数量接种于96孔培养板中，在含5％CO2的湿化培养箱中37℃培养24h；每孔分别加入100μL待测化合物，使最终浓度分别为40.0、20.0、10.0、5.0、2.5、1.25μM。设置细胞对照组及空白对照组，药物组每浓度重复4孔，细胞对照组和空白对照组重复3孔。培养箱中继续培养72h后，每孔加20μL MTT孵育4h，弃去上清，再加100μL DMSO，振荡10min，酶标仪490nm波长测得吸光度值，记录结果，并计算化合物的IC50值，细胞抑制率(％)＝[1－(A给药－A空白)/(A正常－A空白)]×100％。
3.结果
3.1本发明化合物LQC-C对5种肿瘤细胞系(HepG2、HT-29、Hela、BGC-823、A549)和人肝正常细胞L02的IC50值如表1所示。
由表1可以看出，化合物LQC-C对各种癌细胞均表现出较好的抑制肿瘤细胞增殖活性，并且其抗肿瘤活性优于原料；其中，化合物LQC-C的对人结肠癌细胞系HT-29和人宫颈癌细胞系Hela和人肺癌细胞系A549抗增殖活性优于阳 性药顺铂；但对人肝正常细胞L02的细胞毒性明显低于阳性药顺铂。
表1 LQC-C对不同肿瘤细胞株和正常细胞的IC50值
Figure PCTCN2017071276-appb-000003
4.结论
本发明化合物表现出抑制肿瘤细胞系(HepG2、HT-29、Hela、BGC-823、A549)增殖的活性。
具体实施方式
实施例1化合物2-氯代甲基-3,5,6-三甲基吡嗪(化合物5)的制备
取川芎嗪21.76g(160mmol)(化合物1)溶于50ml冰乙酸，加入18ml(160mmol)30％过氧化氢于90℃反应4h，之后补充18ml(160mmol)30％过氧化氢继续反应2h，TLC监测至反应完全，冷却至室温，以50％氢氧化钠调pH至10，二氯甲烷萃取，无水硫酸钠干燥，过滤，回收溶剂，得到白色川芎嗪单氮氧(化合物2)。在化合物2中加入15.1ml(160mmol)乙酸酐，于105℃加热回流2.5h，TLC监测至反应完全后，减压蒸除过量的醋酐，得到黑色浆状川芎嗪乙酰化物(化合物3)。将次此黑色浆状物置于100ml(THF:MeOH:H2O＝3:1:1)的溶液，分批加入19.2g(480mmol)氢氧化钠，搅拌反应2h，二氯甲烷萃取，无水硫酸钠干燥，过滤，减压蒸除溶剂，得到羟基川芎嗪粗品，以正己烷重结晶，得18.5g黄色针状结晶(化合物4)。最后将此黄色结晶溶于150ml无水四氢呋喃中，然后加入30.3g(0.159mol)Tscl、24.64g(0.244mol)TEA、1.5g(0.122mol)DMAP，搅拌过夜，二氯甲烷萃取，无水硫酸钠干燥，过滤，回收溶剂，得到淡黄色氯代川芎嗪粗品，硅胶柱分离[V(石油醚):V(乙酸乙酯)＝10:1]得无色透明的氯代川芎嗪(化合物5)，收率80％。
备实施例2化合物TBA的制备
称取12g(26.28mmol)白桦脂酸，7.25g(52.56mmol)无水碳酸钾于100ml单口瓶中，加入50mlDMF，搅拌半小时后加入5.4g(31.5mmol)氯代川芎嗪(化合物5)，氮气保护下，于60℃反应12h，TLC检测至反应完全，加入一定量的水终止反应，随后用二氯甲烷萃取，收集有机层，适量无水硫酸钠除水，减压蒸干，硅胶柱分离[V(石油醚):V(丙酮)＝10:1]得白色固体TBA，熔点:184.6–185.4℃,收率90％。1H-NMR(CDCl3)(ppm):0.78,0.80,0.82,0.96,0.98,1.69(s,each,3H,6×-CH3),2.51(s,3H,-CH3),2.53(s,3H,-CH3),2.57(s,3H,-CH3),3.02(m,1H),3.19(m,1H),4.61,4.74(each,brs,1H,＝CH2),5.20,5.23(each,d,J＝12.5Hz,1H,-CH2),1.00–2.50(25H,methyl-and methylene-of triterpenoid structure).13C-NMR(CDCl3)(ppm):14.7,15.4,15.9,16.1,18.3,19.4,20.9,25.5,27.4,28.0,29.7,30.6,32.1,34.4,36.9,37.2,38.1,38.7,38.9,40.7,42.4,46.9,49.5,50.6,55.4,56.7,79.0,109.6,150.5,175.5(-COO-)；pyrazine ring:20.4(-CH3),21.4(-CH3),21.6(-CH3),64.3(-CH2),145.4,148.7,148.9,150.9.HRMS(ESI)m/z:591.45212[M+H]+,理论计算C38H58N2O3 590.44474.
实施例3化合物LQC-C的制备
依次称取200mg(0.338mmol)TBA、83.25mg(0.44mmol)Boc-L-肌氨酸、97.20mg(0.50mmol)EDCI和4.12mg(0.0038mmol)DMAP置于50mL单口瓶中，加入5ml的无水二氯甲烷使其溶解，室温下搅拌过夜TLC[V(二氯甲烷):V(甲醇)＝20:1]检测反应进程，反应液用二氯甲烷萃取，收集有机层，适量无水硫酸钠除水，减压蒸干，硅胶柱分离[V(二氯甲烷):V(甲醇)＝40:1]得白色固体；将其溶于含3M HCl的乙酸乙酯中，冰盐浴搅拌1h；减压蒸干反应液，适量乙酸乙酯复溶，用饱和的碳酸氢钠溶液调pH至中性，收集有机层，适量无水硫酸钠除水，减压蒸干，硅胶柱分离[V(二氯甲烷):V(甲醇)＝40:1]得白色固体LQC-C，熔点:172.6–173.4℃，收率:76.90％。1H-NMR(CDCl3)(ppm):0.77,0.84,0.93(s,each,3H,3×-CH3,methyl of BA),0.82(brs,6H,2×-CH3,methyl of BA),1.00-2.50(31H,methyl-and methylene-of BA),2.40,2.50,2.53(s,each,3H,3×-CH3,methyl of TMP),2.96-3.01(m,1H,-CCHCH2-),3.52(brs,2H,-CH2NH-),4.48-4.51(m,1H,-OCOCH-),4.56,4.59(brs,each,1H,＝CH2),5.17,5.20(d,each,1H,J＝15Hz,-OCH2-).13C-NMR(CDCl3)(ppm):14.79,16.00,16.28,16.64,18.27,19.47,20.63, 23.85,25.56,28.22,29.75,30.66,32.15,34.35,34.55,37.04,37.21,38.01,38.18,38.47,40.79,42.53,46.97,49.59,50.58,50.73,55.55,56.79(-CH-NH2),83.23(-OCOCH-),109.81(-CH＝C-),151.10(-CH＝C-),168.63(-COCH-),175.66(-COO-),pyrazine ring:21.02(-CH3),21.51(-CH3),21.80(-CH3),64.48(-CH2),145.46,148.89,148.98,150.60.HRMS(ESI)m/z:[M+H]+662.48975，理论计算：C41H63N3O4 661.48186.
实施例4
取LQC-C 10g，加入注射剂(包括冻干粉针剂和无菌分装干粉针剂)适当辅料，按注射剂(包括冻干粉针剂和无菌分装干粉针剂)工艺制备成抗肿瘤药注射剂。
实施例5
取LQC-C 10g，加入片剂(包括缓控释片、骨架片、包衣片、分散片等)适当辅料，按片剂(包括缓控释片、骨架片、包衣片、分散片等)工艺制备成抗肿瘤药片剂。
实施例6
取LQC-C 10g，加入胶囊剂适当辅料，按胶囊剂工艺制备成抗肿瘤药胶囊剂。
实施例7
取LQC-C 10g，加入乳剂(包括微乳、纳米乳等)适当辅料，按乳剂(包括微乳、纳米乳等)工艺制备成抗肿瘤药乳剂。
实施例8
取LQC-C 10g，加入颗粒剂适当辅料，按颗粒剂工艺制备成抗肿瘤药颗粒剂。
实施例9
取LQC-C 10g，加入缓释控释剂适当辅料，按缓释控释剂工艺制成抗肿瘤药缓释控释剂。
实施例10
取LQC-C 10g，加入口服液适当辅料，按口服液工艺制备成抗肿瘤药口服液。
实施例11
取LQC-C 10g，加入脂质体剂型适当辅料，按脂质体工艺制备成抗肿瘤药脂质体剂型。

Claims (10)

  1. 具有式1结构的化合物或其药学上可接受的盐，其结构式为：
    Figure PCTCN2017071276-appb-100001
  2. 如权利要求1所述的所述化合物或其药学上可接受的盐，其特征在于，加入制剂领域常规辅料制成片剂、胶囊剂、颗粒剂、散剂、口服液、注射剂等常规剂型。
  3. 如权利要求1所述化合物的制备方法，其特征在于，该方法包括如下步骤：
    步骤1，将白桦脂酸溶解于有机溶剂中，与2-氯代甲基-3,5,6-三甲基吡嗪在碱性条件下生成中间体化合物TBA；
    步骤2，中间体化合物TBA在催化剂的作用下与肌氨酸发生反应生成式1化合物。
  4. 如权利要求3所述的制备方法，其特征在于，反应在-20℃至250℃下进行；所述有机溶剂是含有1-20个碳原子的醚、醇、烷烃、芳香烃、酮、卤代烷、酰胺、腈、酯或者它们各种比例的混合物；所用碱为三乙胺或碳酸钾；所述催化剂为1-羟基苯并三唑、1-乙基-3-(3-二甲胺丙基)碳二亚胺盐酸盐、1,3-二环己基碳二亚胺或4-二甲氨基吡啶。
  5. 如权利要求3所述的制备方法，其特征在于，白桦脂酸与氯代川芎嗪的摩尔比为1:0.1～1:10；白桦脂酸与碱的摩尔比为1:0.1～1:10；TBA与肌氨酸的摩尔比为1:0.1～1:10；TBA与催化剂的摩尔比为1:0.1～1:10。
  6. 如权利要求1所述的化合物或其药学上可接受的盐在制备抗癌药物中的应用。
  7. 如权利要求6所述的应用，其特征在于，所述癌症为肝癌、肺癌、结肠癌、宫颈癌或胃癌。
  8. 药物组合物，其特征在于，所述组合物包含以治疗有效量存在的权利要求1化合物或其药学可接受的盐与至少一种药学可接受的赋形剂的混合物。
  9. 如权利要求8所述的组合物，其特征在于，所述组合物还包含至少一种常规抗癌药。
  10. 如权利要求9所述的组合物，其特征在于，所述抗癌药选自环磷酰胺、5-氟尿嘧啶、紫杉醇、阿霉素、依托泊苷、伊立替康、奥沙利铂、顺铂或健择。
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002016395A1 (en) * 2000-08-18 2002-02-28 The Board Of Trustees Of The University Of Illinois Prodrugs of betulinic acid derivatives for the treatment of cancer and hiv
CN102675401A (zh) * 2011-03-09 2012-09-19 雷海民 抗肿瘤药物lqc-y的制备及其应用

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101230082B (zh) * 2006-05-12 2010-12-01 中国药科大学 23-羟基白桦酸类衍生物、其制备方法、制剂及用途
CN101928321B (zh) * 2010-03-02 2012-09-05 福州大学 具抗癌活性的经酸性氨基酸化学修饰的熊果酸衍生物
CN102344482A (zh) * 2011-07-29 2012-02-08 温州大学 桦木醇衍生物、制备方法及用途
CN104548120A (zh) * 2013-10-22 2015-04-29 北京林业大学 聚乙二醇-白桦脂酸结合物及其制备方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002016395A1 (en) * 2000-08-18 2002-02-28 The Board Of Trustees Of The University Of Illinois Prodrugs of betulinic acid derivatives for the treatment of cancer and hiv
CN102675401A (zh) * 2011-03-09 2012-09-19 雷海民 抗肿瘤药物lqc-y的制备及其应用

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BILDZIUKEVICH, ULADZIMIR: "Polyamine derivatives of betulinic acid and β-sitosterol: A comparative investiga", STEROIDS, vol. 100, 8 May 2008 (2008-05-08), pages 27 - 35, XP029135559, ISSN: 0039-128X *
LI, DAN ET AL.: "Recent Advances of Betulinic Acid and Its Derivatives", PROGRESS IN PHARMACEUTICAL SCIENCES, vol. 28, no. 3, 31 December 2004 (2004-12-31), pages 120 - 125 *
XU, BING: "A Series of New Ligustrazine-Triterpenes Derivatives as Anti- Tumor Agents: Design, Synthesis, and Biological Evaluation", INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, vol. 16, no. 9, 2 September 2015 (2015-09-02), pages 21035 - 21055, XP055598469, ISSN: 1422-0067 *
XU, KUO: "Synthesis and biological evaluation of T- OA analogues as the cytotoxic agents", RESEARCH ON CHEMICAL INTERMEDIATES : AN INTERNAT. JOURNAL, vol. 41, no. 9, 22 July 2014 (2014-07-22), pages 6257 - 6269, XP035524937, DOI: doi:10.1007/s11164-014-1737-z *

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