CN114805469A - 含吡唑的白桦脂酸类衍生物,其制备方法及用途 - Google Patents
含吡唑的白桦脂酸类衍生物,其制备方法及用途 Download PDFInfo
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Abstract
本发明涉及一类含吡唑的白桦脂酸类衍生物,其制备方法和用途,具体地,本发明提供了一类具有如下通式I所示结构的化合物,其中,各基团的定义如说明书中所述。本发明通过碘乙酸钠诱导大鼠关节炎模型,观察关节疼痛程度的变化;采用体外诱导分化体系,观察破骨细胞前体发生破骨分化及炎症因子产生水平。结果显示,本发明采用的化合物对破骨细胞分化和炎症因子产生具有抑制活性,对骨关节炎及以骨破坏、骨侵蚀及滑膜炎症为病理特征的关节炎症疾病有良好的治疗作用,具有理想的临床应用前景。
Description
技术领域
本发明属于药物化学和药物治疗领域。具体涉及一类含吡唑的白桦脂酸类衍生物,及其药学上可接受的盐及水合物,其制备方法及它们在治疗破骨细胞活性异常相关疾病的药物的用途。
背景技术
骨质疏松症(osteoporosis,OP)是以骨量减少和骨组织微结构退化为特征,导致骨密度下降、脆性增加而易于发生骨折的系统性骨病。骨质疏松症的产生与骨重建过程中骨吸收与骨形成的失衡有关。人体的骨骼不断地进行新陈代谢,破骨细胞(osteoclast,OC)促进骨吸收,成骨细胞(osteoblast,OB)促进骨形成。破骨细胞被激活,分化成熟后迁移并粘附骨表面进行骨吸收,从而导致旧的骨质被分解。成骨细胞促进新骨的形成,当它被活化、转移至被吸收的骨组织部位时,能通过分泌骨基质而促进钙的沉积,形成新的骨质。(Wu,S等,Genome Medicine.2013,5:44.)当破骨细胞降解骨基质的速度快于成骨细胞重建骨时,这种骨重建平衡就会被破坏,发生骨代谢异常,导致骨质疏松症的发生。此外,激素因子也会影响骨吸收,缺乏雌激素会增加骨吸收,并减少新骨的沉积。钙和维生素D缺乏也会导致骨沉积受损。甲状旁腺素(parathyroid hormone,PTH)是参与骨代谢及骨转换的重要激素之一,可调节骨骼的合成和分解代谢,对成骨细胞分化、成熟及凋亡均发挥重要作用。降钙素是一种由甲状腺产生的增加骨沉积的激素,其作用不如PTH明显。(Khosla S等,Endocrinology 2001,142(12):5050-5055.)破骨细胞的活化受各种分子信号的调节,其中RANKL(Receptor Activator for Nuclear Factor-κB Ligand,NF-κB受体活化因子配体)是重要信号分子之一。该分子由成骨细胞产生,与破骨细胞前体细胞表面的RANK受体结合,促使破骨细胞前体分化,形成成熟的破骨细胞。(Khosla S等,Endocrinology 2001,142(12):5050-5055.)
药物治疗是骨质疏松症的主要治疗方法,目前可用于治疗骨质疏松的药物中,以抑制破骨细胞分化、成熟的骨吸收抑制剂为主,在市场中占主导地位的是双膦酸盐药物、降钙素、选择性雌激素受体调节剂等。激素类药物雌激素可能增加乳腺癌、中风、子宫内膜癌变等风险,一般连续使用期限不超过3年。双膦酸盐长期使用有股骨骨折和颌骨坏死风险。选择性雌激素受体调节剂雷洛昔芬,增加了血栓和中风的风险。(Yasothan U等,Nat.Rev.drug.Discov.2008,7:725-726.)综上所述,这些临床应用的药物均有明显副作用,因此,寻找更为安全有效的治疗骨质疏松新药是抗骨质疏松药物研发的一个重要方向。
骨关节炎(osteoarthritis,OA),指由多种因素引起关节软骨纤维化、皲裂、溃疡、脱失而导致的关节疾病。病因尚不明确,病理特点为关节软骨变性破坏、软骨下骨硬化或囊性病变、关节边缘骨质增生、滑膜增生、关节囊挛缩、韧带松弛或挛缩、肌肉萎缩无力等。OA以中老年患者多见,60岁以上的人群中患病率可达50%,75岁的人群则达80%。该病的致残率可高达53%。(中华医学会骨科学分会,Orthopedic Journal of China,2014,22(03):287-288.)
OA的治疗目的是减轻疼痛,矫正畸形,改善生活质量,总体治疗原则是非药物与药物治疗相结合,必要时手术治疗,药物治疗分为改善症状和改变病情的药物两类。
改善OA症状类药物以止痛药为主,首选对乙酰氨基酚,但长期大剂量使用有引起肝或肾损伤的报告。非甾体抗炎药具有抗炎、止痛和解热作用,可用于治疗OA,使用前应权衡患者胃肠道、肝、肾、心血管疾病风险,不宜长期使用;中度至严重的膝OA患者,可用阿片类药物,但应注意其不良反应及成瘾性。改善OA病情的代表性药物是硫酸氨基葡萄糖,作用机理可能与软骨代谢相关。此外,还有关节腔注射给药,常用药物包括糖皮质激素、几丁糖、玻璃酸钠等,可有效缓解疼痛,改善关节功能。
与其他治疗方法相比,药物治疗骨关节炎简便易行、疗效可靠,但现有药物均存在明显不足,值得进一步探索。
天然产物是创新药物的摇篮,通过对天然产物进行结构修饰,是新药发现的重要途径之一。白桦脂酸(Betulinic acid)属于五环三萜类羽扇豆烷型化合物,是从天然产物中提取的活性化合物,其衍生物有多种生物活性——抗HIV、抗癌、抗病毒、抗炎症、抗氧化等,毒性低、安全指数大。可以抑制RANKL诱导RAW264.7分化形成成熟破骨细胞,抑制破骨细胞成熟(Li C等,J.Bone.Miner.Res.2011.26(3):644-656.),还可以通过激活BMP/Smad/Runx2与β-catenin信号通路及上调OPG/RANKL比例,在一定程度上促进成骨细胞分化成熟。
虽然白桦脂酸具有多种潜在生物活性,但其也有溶解度低、logP值过大等问题,导致生物利用度偏低。水溶性差是限制白桦脂酸应用的一个主要原因,现有的研究对白桦脂酸进行了一定的结构改造,以寻找具有更高活性和更好理化性质的类似物。白桦脂酸活性位点较多,结构稍作修改,活性即有很大改变。现有文献报道了在C-3、C-28、C-29等位置做结构修饰,其衍生物的生物活性在抗肿瘤、抗病毒、抗炎、抗HIV等方面均有很大的提高。
现有研究发现,2,3位吡唑并白桦脂酸衍生物抑制破骨细胞成熟活性相对于白桦脂酸提高了近200倍,细胞毒实验显示,吡唑并白桦脂酸衍生物抑制破骨细胞分化的活性并非由细胞毒性引起。动物试验证明,吡唑并白桦脂酸衍生物能有效抑制骨质疏松模型鼠骨质疏松症的发生,但其溶解度较差,水中的溶解度小于l mg/L,大多数有机溶剂中溶解性也不佳,溶解度低导致生物利用度差是该类化合物的主要问题。(Xu,J等,J.Med.Chem.2012,55(7):3122-3134.)
综上所述,本领域需开发新的具有更好破骨细胞分化抑制活性、炎症因子抑制活性,并具有优良理化性质和安全性的吡唑并白桦脂酸衍生物。
发明内容
本发明人经过长期而深入的研究,发现了一类C-17位胺基取代的吡唑并白桦脂酸类化合物,所述化合物可以抑制破骨细胞的活性,从而用于与破骨细胞活性失衡有关的骨代谢疾病。基于上述发现,发明人完成了本发明。
本公开的一个目的是提供一种吡唑并白桦脂酸衍生物。
本公开的另一个目的是提供所述吡唑并白桦脂酸衍生物的制备方法。
本公开的又一个目的是提供包括所述吡唑并白桦脂酸衍生物的药物组合物。
本公开的再一个目的是提供所述吡唑并白桦脂酸衍生物或包含其的药物组合物在制备治疗与破骨细胞活性抑制、免疫活性抑制、炎症因子活性抑制相关的疾病的药物中的用途。
根据本公开的第一方面,其提供了一种如通式I中所述的基于吡唑并白桦脂酸的化合物,其异构体及药学上可接受的盐:
I
其中,
R1选自以下组:异丙基、异丙烯基;
R2选自以下组:氢、取代或未取代的C1-C4直链或支链烷基;
R3选自以下组:氢、取代或未取代的C1-C4直链或支链烷基、取代或未取代的C3-C6环烷基、取代或未取代的C1-C4直链或支链烷氧基、取代或未取代的C3-C6的环烷基氧基、取代或未取代的C1-C4直链或支链烷基氨基、取代或未取代的含氧或氮或硫的饱和五元或六元杂环基或其氧化物、取代或未取代的含氧或氮或硫的不饱和五元或六元杂环基;
X不存在,或者选自以下组:亚甲基、羰基、-CONH-;
n为0-4的整数;
其中,所述的取代指被选自以下组的取代基取代:卤素、C1-C8直链或支链烷基、C1-C8直链或支链烷基氨基(例如甲氨基)、C1-C8直链或支链烷氧羰基(例如叔丁氧羰基)、羟基、-NH2、C1-C8直链或支链烷氧基、氧代基团(=O)。
根据本公开的第二方面,提供了一种基于吡唑并白桦脂酸的化合物制备方法,包括:
方法一:
或者
方法二:
步骤(a1)如上述方法一;
(a2)将Ia1还原以制备式Ia2化合物:
或者
方法三:
或者
方法四:
式中,Y选自以下组:氯、溴、碘、甲磺酸酯基、对甲苯磺酸酯基、三氟甲磺酸酯基等离去基团;其余各基团定义如上文所述。
根据本公开的第三方面,其提供了一种药物组合物,所述的药物组合物含有治疗有效量的选自如本发明第一方面所述的化合物、其异构体及药学上可接受的盐中的一种或多种。
根据本公开的第四方面,其提供了如本发明第一方面所述的化合物、其异构体及药学上可接受的盐或如本发明第三方面所述的药物组合物在制备治疗与破骨细胞活性抑制、免疫活性抑制、炎症因子活性抑制相关的疾病的药物中的用途。
根据本公开的第五方面,其提供了一种抑制破骨细胞活性、免疫活性、或炎症因子活性的方法,所述方法包括步骤:给需要的对象施用治疗有效量的选自如本发明第一方面中所述的化合物、其异构体及药学上可接受的盐中的一种或多种或如本发明第三方面所述的药物组合物。
有益效果
与现有技术相比,本发明的主要优点包括:
1.提供了一类结构新颖的化合物,所述化合物具有破骨细胞和炎症因子抑制活性。
2.提供了一类具有破骨细胞抑制活性的化合物,可以用于制备治疗骨代谢相关疾病的药物。
附图说明
图1是显示出本公开的部分化合物在不同浓度下对白介素-1β的抑制率的图。
图2是显示出正常组、模型组、不同剂量的I-11化合物的治疗组、地塞米松对照治疗组中碘酸钠注射造模后若干天后的50%缩足痛阈示意图。
图3是显示出正常组、模型组、不同剂量的I-11化合物的治疗组、地塞米松对照治疗组中碘酸钠注射造模后若干天后的左足承重占比的示意图。
具体实施方式
为使本领域具有普通知识的人员可了解本发明的特点及效果,以下谨就说明书及申请专利范围中提及的术语及用语进行一般性的说明及定义。除非另有指明,否则文中使用的所有技术及科学上的字词,皆具有本领域技术人员对于本发明所了解的通常意义,当有冲突情形时,应以本说明书的定义为准。
在本文中,用语“包含”、“包括”、“具有”、“含有”或其他任何类似用语均属于开放性连接词(open-ended transitional phrase),其意欲涵盖非排他性的包括物。举例而言,含有复数要素的一组合物或制品并不仅限于本文所列出的这些要素而已,而是还可包括未明确列出但却是该组合物或制品通常固有的其他要素。除此之外,除非有相反的明确说明,否则用语“或”是指涵盖性的“或”,而不是指排他性的“或”。例如,以下任何一种情况均满足条件“A或B”:A为真(或存在)且B为伪(或不存在)、A为伪(或不存在)且B为真(或存在)、A和B均为真(或存在)。此外,在本文中,用语“包含”、“包括”、“具有”、“含有”的解读应视为已具体公开并同时涵盖“由…所组成”及“实质上由…所组成”等封闭式或半封闭式连接词。
在本文中,所有以数值范围或百分比范围形式界定的特征或条件仅是为了简洁及方便。据此,数值范围或百分比范围的描述应视为已涵盖且具体公开所有可能的次级范围及范围内的个别数值,特别是整数数值。举例而言,“1至8”的范围描述应视为已经具体公开如1至7、2至8、2至6、3至6、4至8、3至8等等所有次级范围,特别是由所有整数数值所界定的次级范围,且应视为已经具体公开范围内如1、2、3、4、5、6、7、8等个别数值。除非另有指明,否则前述解释方法适用于本发明全文的所有内容,不论范围广泛与否。
若数量或其他数值或参数是以范围、较佳范围或一系列上限与下限表示,则其应理解成是本文已特定公开了由任一对该范围的上限或较佳值与该范围的下限或较佳值构成的所有范围,不论这些范围是否有分别公开。此外,本文中若提到数值的范围时,除非另有说明,否则该范围应包括其端点以及范围内的所有整数与分数。
在本文中,在可实现发明目的的前提下,数值应理解成具有该数值有效位数的精确度。举例来说,数字40.0则应理解成涵盖从39.50至40.49的范围。
在本文中,对于使用马库什群组(Markush group)或选项式用语以描述本发明特征或实例的情形,本领域技术人员应了解马库什群组或选项列表内所有要素的次级群组或任何个别要素亦可用于描述本发明。举例而言,若X描述成“选自于由X1、X2及X3所组成的群组”,亦表示已经完全描述出X为X1的主张与X为X1及/或X2的主张。再者,对于使用马库什群组或选项式用语以描述本发明的特征或实例的情况,本领域技术人员应了解马库什群组或选项列表内所有要素的次级群组或个别要素的任何组合亦可用于描述本发明。据此,举例而言,若X描述成“选自于由X1、X2及X3所组成的群组”,且Y描述成“选自于由Y1、Y2及Y3所组成的群组”,则表示已经完全描述出X为X1或X2或X3而Y为Y1或Y2或Y3的主张。
术语
除特别说明之处,本发明的所有化合物之中,各手性碳原子可以任选为R构型或S构型,或R构型和S构型的混合物。
术语“C1-C4烷基”指具有1~4个碳原子的直链或支链烷基,例如甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、叔丁基、或类似基团。
术语“C3-C6环烷基”指具有3~6个碳原子的环烷基,例如环丙基、环丁基、环戊基、环己基、或类似基团。
术语“C1-C4烷氧基”指具有1~4个碳原子的烷氧基,例如甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、异丁氧基、仲丁氧基、叔丁氧基、或类似基团。
术语卤素指F、Cl、Br和I。
术语“饱和五元或六元杂环基”指环上包含选自氧、氮、硫的至少一个杂原子但不含有不饱和键的五元或六元杂环基,例如I-5。
术语“不饱和五元或六元杂环基”指环上包含选自氧、氮、硫的至少一个杂原子且包含至少一个不饱和键的五元或六元杂环基,例如I-39。
术语“取代或未取代的含氧或氮或硫的饱和五元或六元杂环基或其氧化物”中“氧化物”是指杂环环原子中的碳、氮或硫被氧化所得到的产物,例如硫被氧化形成如-S(=O)2-或-S(=O)-的结构,碳被氧化形成如-C(=O)-的结构,氮被氧化形成如-N+(-O-)-的结构。
以下具体实施方式本质上仅是例示性,且并不欲限制本发明及其用途。此外,本文并不受前述现有技术或发明内容或以下具体实施方式或实施例中所描述的任何理论的限制。
根据本公开的一个实施方式,其提供了一种如通式I中所述的基于吡唑并白桦脂酸的化合物,其异构体及药学上可接受的盐:
I
其中,
R1选自以下组:异丙基、异丙烯基;
R2选自以下组:氢、取代或未取代的C1-C4直链、支链或环状烷基;
R3选自以下组:氢、取代或未取代的C1-C4直链或支链烷基、取代或未取代的C3-C6的环烷基、取代或未取代的C1-C4直链或支链烷氧基、取代或未取代的C3-C6的环烷基氧基、取代或未取代的C1-C4直链或支链烷基氨基、取代或未取代的含氧或氮或硫的饱和五元或六元杂环基或其氧化物、取代或未取代的含氧或氮或硫的不饱和五元或六元杂环基;
X不存在,或者选自以下组:亚甲基、羰基、-CONH-;
n为0-4的整数;
其中,所述的取代指被选自以下组的取代基取代:卤素、C1-C8直链或支链烷基、C1-C8直链或支链烷基氨基(例如甲氨基)、C1-C8直链或支链烷氧羰基(例如叔丁氧羰基)、羟基、-NH2、C1-C8直链或支链烷氧基、氧代基团(=O)。
在一些实施方式中,R2选自以下组:氢、甲基。
在一些实施方式中,R3选自以下组:氢、甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基、环丁基、环戊基、环已基、甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、仲丁氧基、异丁氧基、叔丁氧基、环丁基氧基、环戊基氧基、环已基氧基、-NH2、
在一些实施方式中,X选自下组:亚甲基、羰基、-CONH-。
n为0、1、2、3或4。
根据本公开的一个实施方式,其中,所述的式I化合物具有选自下组的结构:
根据本公开的一个实施方式,其中,
所述药学可接受的盐包括有机酸盐和无机酸盐,包括但不限于马来酸盐、琥珀酸盐、柠檬酸盐、酒石酸盐、富马酸盐、乙酸盐、甲磺酸盐、盐酸盐、磷酸盐、硝酸盐或硫酸盐。
根据本公开的一个实施方式,其提供了一种基于吡唑并白桦脂酸的化合物的制备方法,包括:
方法一:
或者
方法二:
步骤(a1)如上述方法一;
(a2)将Ia1还原以制备式Ia2化合物:
或者
方法三:
方法四:
式中,Y为离去基团,选自以下组:氯、溴、碘、甲磺酸酯基、对甲苯磺酸酯基、三氟甲磺酸酯基等;其余各基团定义如上文所述。
根据本公开的一个实施方式,化合物6如下制备:
化合物1与2-碘酰基苯甲酸(IBX)反应得化合物2;
化合物2在甲醇钠存在下与甲酸乙酯反应得化合物3;
化合物3与二盐酸肼反应得化合物4;
化合物4在三乙胺(TEA)存在下与叠氮磷酸二苯酯(DPPA)反应得化合物5;
化合物5水解得化合物6。
根据本公开的一个实施方式,所述步骤(a1)中,在有机溶剂中,使化合物6与侧链化合物在碱的作用下,进行取代反应生成化合物7,然后化合物7与侧链化合物R2-Y,在碱的作用下,进一步进行取代反应生成化合物Ia1。
根据本公开的一个实施方式,所述步骤中的碱为N,N-二异丙基乙胺(DIPEA)或三乙胺(TEA)。
根据本公开的一个实施方式,所述的式6化合物与N,N-二异丙基乙胺(DIPEA)或三乙胺(TEA)的摩尔比为1:1.4-2.0。
根据本公开的一个实施方式,所述的式6化合物与侧链化合物的摩尔比为1:1.4-2.0。
根据本公开的一个实施方式,在所述的步骤(a1)中,所述的有机溶剂选自下组:四氢呋喃、吡啶、N,N-二甲基甲酰胺、二氯甲烷、1,2-二氯乙烷、氯仿、二氧六环,或其组合。
根据本公开的一个实施方式,在所述的步骤(a1)中,反应温度为40-120℃。
根据本公开的一个实施方式,所述步骤(a2)中,化合物Ia1在有机溶剂中,在还原剂作用下还原成化合物Ia2。优选地,还原条件为钯碳(10%)+氢气还原。
根据本公开的一个实施方式,钯碳用量为20%。
根据本公开的一个实施方式,反应压力为100kPa。
根据本公开的一个实施方式,在所述步骤(a2)中,所述有机溶剂选自下组:四氢呋喃、甲醇、二氯甲烷,或其组合。
根据本公开的一个实施方式,在所述步骤(a2)中,反应温度为20-40℃。
根据本公开的一个实施方式,所述的步骤(b)在TEA和/或4-二甲氨基吡啶(DMAP)的存在下进行。
根据本公开的一个实施方式,所述的式6化合物与TEA的摩尔比为1:2.0-3.0。
根据本公开的一个实施方式,所述的式6化合物与DMAP的摩尔比为1:0.1-0.2。
根据本公开的一个实施方式,在所述的步骤(b)在有机溶剂中进行,所述的有机溶剂选自下组:二氯甲烷、四氢呋喃、二氧六环、1,2-二氯乙烷、N,N-二甲基甲酰胺,或其组合。
根据本公开的一个实施方式,在所述的步骤(b)中,反应温度为20-40℃。
根据本公开的一个实施方式,所述的步骤(c)在DIPEA存在下进行。
根据本公开的一个实施方式,所述的步骤(c)中,式5化合物与DIPEA的摩尔比为1:1.4-2.4。
根据本公开的一个实施方式,在所述的步骤(c)在有机溶剂中进行,所述的有机溶剂选自下组:二氧六环、四氢呋喃、二氯甲烷,或其组合。
根据本公开的一个实施方式,在所述的步骤(c)中,反应温度为20-40℃。
根据本公开的一个实施方式,其提供了一种药物组合物,所述的药物组合物含有治疗有效量的选自如本发明第一方面所述的化合物、其异构体及药学上可接受的盐中的一种或多种。所述的药物组合物还可以(任选地)包括药学上可接受的辅料。
根据本公开的一个实施方式,其提供了如本发明第一方面所述的化合物、其异构体及药学上可接受的盐或如本发明第三方面所述的药物组合物在制备治疗与破骨细胞活性抑制、免疫活性抑制、炎症因子活性抑制相关的疾病的药物中的用途。
根据本公开的一个实施方式,其提供了一种抑制破骨细胞活性、免疫活性、或炎症因子活性的方法,所述方法包括:给需要的对象施用治疗有效量的选自如本发明第一方面中所述的化合物、其异构体及药学上可接受的盐中的一种或多种或如本发明第三方面所述的药物组合物。
根据本公开的一个实施方式,所述与破骨细胞活性抑制、免疫活性抑制、炎症因子活性抑制相关的疾病包括骨质疏松症、骨关节炎、牙周炎、牙齿脱落、Paget’s骨病、佝偻病、骨巨细胞瘤、骨髓瘤骨病以及癌症骨转移造成的骨破坏等。
下面结合具体实施例,进一步阐述本发明。应理解,这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的试验方法,通常按照常规条件,或按照制造产商所建议的条件。除非另外说明,否则百分比和份数按重量计算。
下面结合实施例对本发明所涉及化合物的结构和制备方法及体外抑制破骨细胞活性、抗炎症因子作用作进一步阐述,但不限制本发明。
应理解,在本发明范围内中,本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合,从而构成新的或优选的技术方案。限于篇幅,在此不再一一赘述。
所有实施例中,起始化合物1与相关侧链购于韶远科技(上海)有限公司、北京百灵威科技有限公司、上海泰坦科技有限公司。除特殊说明外,其他起始材料、溶剂、材料均来源于国药试剂公司。1H NMR由BrucherAM-400或GEMINI-400型核磁共振仪记录,化学位移以δ(ppm)表示。质谱由Agilent1200-6110型单四级杆液相色谱质谱联用仪记录。分离用200-300目硅胶由青岛海洋化工厂提供。其中英文缩写所代表的化学试剂如下:
DMF N,N-二甲基甲酰胺
THF 四氢呋喃
DCM 二氯甲烷
TMS 三甲基硅基
DIPEA N,N-二异丙基乙胺
TEA三乙胺
DMAP 4-二甲氨基吡啶
化合物的制备与合成
化合物2
于250mL圆底烧瓶中,加入500mg(1.1mmol)化合物1,溶于150mL二甲亚砜中,加入616mg(2.2mmol)2-碘酰基苯甲酸,室温反应8h。反应液倒入冰水:乙酸乙酯(200mL:100mL)混合物中,抽滤,滤液加150mL水,用200mL乙酸乙酯萃取三次,合并有机相,饱和食盐水洗涤,无水硫酸钠干燥,减压蒸干溶剂,粗品用硅胶柱层析纯化(石油醚:乙酸乙酯=7:1),得米白色固体,收率83%。
1H NMR(400MHz,DMSO-d6)δ12.08(s,1H),4.82(dt,J=2.2,1.0Hz,1H),4.72(dt,J=2.4,1.2Hz,1H),2.40–2.19(m,3H),2.15–1.98(m,2H),1.86–1.62(m,6H),1.62–1.08(m,17H),0.97(d,J=3.2Hz,6H),0.92(s,3H),0.83(s,3H),0.77(s,3H).
LC-MS:455.7(M+1)
化合物3
于250mL两颈瓶中,加入300mg(0.66mmol)化合物2,361mg(6.6mmol)甲醇钠,氩气保护,加入150mL超干THF溶解,0.11mL(1.32mmol)甲酸乙酯,室温反应过夜。加入至150mL水中,1M稀盐酸调pH值至2~4,100mL乙酸乙酯萃取三次,合并有机相,饱和食盐水洗涤,无水硫酸钠干燥,减压蒸干溶剂,粗品用硅胶柱层析纯化(石油醚:乙酸乙酯=4:1),得浅黄色固体,收率56%。
1H NMR(400MHz,DMSO-d6)δ12.11(s,1H),9.71(s,1H),4.87(dt,J=2.1,1.1Hz,1H),4.74(dt,J=2.3,1.2Hz,1H),3.41(td,J=7.0,6.1Hz,1H),2.31–2.18(m,2H),2.08(dd,J=13.0,7.0Hz,1H),1.88–1.79(m,2H),1.78–1.64(m,3H),1.63–1.08(m,17H),1.03(q,J=6.9Hz,1H),0.97–0.84(m,12H),0.73(s,3H).
LC-MS:483.7(M+1)
化合物4
于100mL圆底烧瓶中,加入100mg(0.21mmol)化合物3,溶于40mL无水乙醇中,加入51mg(0.48mmol)二盐酸肼,回流5h。冷却至室温,倒入100mL水中,60mL乙酸乙酯萃取三次,合并有机相,饱和食盐水洗涤,无水硫酸钠干燥,减压蒸干溶剂,粗品用硅胶柱层析纯化(石油醚:乙酸乙酯=1:1),得浅黄色固体,收率74%。
1H NMR(400MHz,DMSO-d6)δ12.20(s,2H),7.11(s,1H),4.71(d,J=2.5Hz,1H),4.58(s,1H),2.95(dd,J=10.8,5.4Hz,1H),2.54(s,1H),2.28(t,J=12.4Hz,1H),2.12(d,J=9.3Hz,1H),2.00(q,J=7.0,6.4Hz,1H),1.90–1.73(m,2H),1.66(s,3H),1.58–1.12(m,18H),1.08(s,3H),0.96(s,3H),0.92(s,3H),0.85(t,J=6.4Hz,1H),0.71(s,3H).
LC-MS:479.7(M+1)
化合物5
于100mL圆底烧瓶中,加入100mg(0.21mmol)化合物4,溶于50mL的1,4-二氧六环中,加入87μL(0.63mmol)三乙胺,94μL(0.42mmol)叠氮磷酸二苯酯,氩气保护,回流过夜。冷却至室温,倒入100mL水中,50mL二氯甲烷萃取三次,合并有机相,饱和食盐水洗涤,无水硫酸钠干燥,减压蒸干溶剂,粗品用硅胶柱层析纯化(二氯甲烷:甲醇=20:1),得浅黄色固体,收率63%。
1H NMR(400MHz,DMSO-d6)δ12.41(s,1H),7.02(s,1H),4.98–4.91(m,1H),4.80(dd,J=2.2,1.0Hz,1H),2.52(d,J=13.0Hz,1H),2.36–2.23(m,2H),1.83–1.74(m,1H),1.70–1.24(m,25H),1.20–1.02(m,3H),0.98(s,3H),0.82(s,3H),0.71(s,3H).
LC-MS:476.7(M+1)
化合物6/I-1
于100mL圆底烧瓶中,加入50mg(0.11mmol)化合物5,溶解于50mL的1,4-二氧六环中,加入50μl浓盐酸,60℃反应16h。冷却至室温,减压蒸干,二氯甲烷-甲醇混合液(20mL:20mL)溶解,减压蒸干,重复2~3次,得浅棕色盐酸盐固体,产率91%。
1H NMR(400MHz,DMSO-d6)δ12.48(s,1H),6.97(s,1H),4.94(dt,J=2.1,1.2Hz,1H),4.79(dt,J=2.2,1.0Hz,1H),2.50(d,J=12.9Hz,1H),2.33(d,J=12.9Hz,1H),2.21(tt,J=7.0,1.1Hz,1H),1.78(s,2H),1.69–1.43(m,12H),1.42–0.99(m,15H),0.97(s,3H),0.86(d,J=9.5Hz,5H),0.69(s,3H).
LC-MS:450.7(M+1)
化合物I-2
于100mL圆底烧瓶中,加入100mg(0.22mmol)化合物6,溶解于50mL二氯甲烷中,加入74μL(0.44mmol)DIPEA,31μL(0.14mmol)甲磺酸甲酯,回流48h。冷却至室温,加入150mL水中,50mL二氯甲烷萃取三次,合并有机相,饱和食盐水洗涤,无水硫酸钠干燥,减压蒸干,粗品用硅胶柱层析纯化(二氯甲烷:甲醇=10:1),得浅黄色固体,收率13%。
1H NMR(400MHz,CDCl3)δ6.89(s,1H),4.86(dt,J=2.3,1.2Hz,1H),4.71(dt,J=2.4,1.2Hz,1H),2.54(d,J=12.9Hz,1H),2.46–2.34(m,4H),2.23(dt,J=6.9,1.1Hz,1H),1.82–1.06(m,28H),1.03(s,3H),0.91(s,4H),0.75(s,3H).
LC-MS:464.7(M+1)
化合物I-3
以化合物6为原料,参照化合物I-2的合成过程,柱层析分离得化合物I-2和化合物I-3。化合物I-3为棕色固体,收率17%。
1H NMR(400MHz,CDCl3)δ6.88(s,1H),4.83(dt,J=2.5,1.2Hz,1H),4.73(dt,J=2.5,1.2Hz,1H),2.52(d,J=13.0Hz,1H),2.36(d,J=12.9Hz,1H),2.28(s,7H),1.79–1.00(m,32H),0.92(s,3H),0.73(s,3H).
LC-MS:478.8(M+1)
化合物I-4
以化合物6为原料,参照化合物I-2的合成过程,将甲磺酸甲酯替换为甲磺酸丙酯。得浅棕色固体,收率8%。
1H NMR(400MHz,CDCl3)δ6.79(s,1H),4.82(dt,J=2.2,1.1Hz,1H),4.77–4.66(m,1H),2.98–2.84(m,1H),2.73(ddd,J=12.4,10.8,4.7Hz,1H),2.54(d,J=12.9Hz,1H),2.37(d,J=13.0Hz,1H),2.33–2.19(m,1H),1.79–1.08(m,30H),1.03(s,3H),0.98–0.87(m,7H),0.75(s,3H).
LC-MS:492.8(M+1)
化合物I-5
1H NMR(400MHz,CDCl3)δ6.88(s,1H),4.80(dt,J=2.3,1.1Hz,1H),4.68(dt,J=2.4,1.2Hz,1H),2.89(dt,J=12.5,7.1Hz,2H),2.59–2.45(m,2H),2.44–2.32(m,4H),2.22–2.09(m,3H),1.83–1.06(m,32H),1.04(s,3H),0.94–0.86(m,4H),0.74(s,3H).
LC-MS:547.9(M+1)
化合物I-6
1H NMR(400MHz,CDCl3)δ6.81(s,1H),4.75(dt,J=2.2,1.1Hz,1H),4.71–4.65(m,1H),2.54–2.41(m,3H),2.31(d,J=12.9Hz,1H),2.02(tt,J=7.0,1.1Hz,1H),1.83–1.27(m,28H),1.26–0.98(m,13H),0.79(s,3H),0.74(s,3H).
LC-MS:532.9(M+1)
化合物I-7
1H NMR(400MHz,CDCl3)δ6.91(s,1H),4.83(dt,J=2.2,1.1Hz,1H),4.73(dt,J=2.4,1.1Hz,1H),3.99(t,J=7.0Hz,1H),3.89–3.80(m,1H),3.75(dt,J=8.0,7.0Hz,1H),2.99(dd,J=12.3,7.0Hz,1H),2.83(dd,J=12.5,7.0Hz,1H),2.49(d,J=13.0Hz,1H),2.32(d,J=13.1Hz,1H),2.29–2.19(m,1H),2.16–2.08(m,1H),2.07–1.98(m,1H),1.84–1.61(m,9H),1.58–1.03(m,25H),0.77(d,J=7.9Hz,6H).
LC-MS:534.8(M+1)
化合物I-8
1H NMR(400MHz,CDCl3)δ6.93(s,1H),4.85(s,1H),4.76(s,1H),3.20(d,J=12.5Hz,1H),2.92(dt,J=12.5,7.1Hz,1H),2.74(d,J=12.5Hz,1H),2.49(d,J=12.9Hz,1H),2.36–2.29(m,4H),2.25(s,1H),2.15–1.99(m,2H),1.84–1.57(m,11H),1.58–1.05(m,27H),0.76(d,J=7.0Hz,6H).
LC-MS:561.9(M+1)
化合物I-9
1H NMR(400MHz,CDCl3)δ6.93(s,1H),4.82(dt,J=2.1,1.2Hz,1H),4.76(dt,J=2.2,1.1Hz,1H),2.55(d,J=13.1Hz,1H),2.47(d,J=6.9Hz,2H),2.36–2.18(m,2H),1.89–0.92(m,43H),0.78(d,J=4.5Hz,6H).
LC-MS:546.9(M+1)
化合物I-10
1H NMR(400MHz,CDCl3)δ6.89(s,1H),4.84(dt,J=2.2,1.2Hz,1H),4.75(dt,J=2.1,1.1Hz,1H),3.09(dt,J=9.5,7.0Hz,1H),2.98(dd,J=12.4,7.0Hz,1H),2.82(dd,J=12.4,7.0Hz,1H),2.61(d,J=13.0Hz,1H),2.54(q,J=7.0Hz,1H),2.32(d,J=10.0Hz,4H),2.27–2.18(m,2H),2.03(dt,J=12.9,7.0Hz,1H),1.85(dq,J=12.9,7.0Hz,1H),1.78–1.44(m,15H),1.43–0.99(m,18H),0.94(q,J=7.0Hz,1H),0.86(s,3H),0.77(s,3H).
LC-MS:547.9(M+1)
化合物I-11
1H NMR(400MHz,CDCl3)δ6.86(s,1H),4.87(dt,J=2.2,1.1Hz,1H),4.77(dt,J=2.2,1.1Hz,1H),3.10(dt,J=12.5,7.1Hz,2H),2.54(d,J=13.1Hz,1H),2.47(d,J=6.9Hz,2H),2.37(d,J=13.1Hz,1H),2.32–2.19(m,6H),1.78–1.27(m,28H),1.25–1.06(m,5H),1.04(s,3H),0.95–0.85(m,4H),0.75(s,3H).
LC-MS:561.9(M+1)
化合物I-12
1H NMR(400MHz,CDCl3)δ6.91(s,1H),4.85(dt,J=2.4,1.2Hz,1H),4.79(dt,J=2.4,1.2Hz,1H),2.87(dt,J=12.5,7.1Hz,1H),2.66(dt,J=12.5,7.1Hz,1H),2.49(d,J=12.9Hz,1H),2.31(d,J=13.1Hz,1H),2.24(qt,J=7.1,1.1Hz,1H),1.84–1.60(m,8H),1.58–0.96(m,27H),0.76(d,J=5.3Hz,6H),0.49–0.39(m,2H),0.32–0.23(m,2H).
LC-MS:518.9(M+1)
化合物I-13
1H NMR(400MHz,CDCl3)δ6.79(s,1H),4.79(s,1H),4.73(s,1H),3.10–3.00(m,2H),2.95–2.73(m,2H),2.49(d,J=12.9Hz,1H),2.31(d,J=13.0Hz,1H),2.22(d,J=17.1Hz,4H),2.01(ddt,J=9.5,8.1,7.1Hz,2H),1.90(d,J=12.3Hz,1H),1.86–1.59(m,11H),1.59–1.25(m,15H),1.23–1.03(m,10H),0.77(d,J=9.8Hz,6H).
LC-MS:561.9(M+1)
化合物I-14
1H NMR(400MHz,CDCl3)δ7.56(s,1H),7.04(d,J=7.4Hz,1H),6.96–6.84(m,2H),4.84(dt,J=2.1,1.2Hz,1H),4.73(dt,J=2.2,1.1Hz,1H),4.10(ddd,J=22.7,11.6,3.5Hz,2H),3.63–3.39(m,1H),3.23–3.00(m,1H),2.54(d,J=13.0Hz,1H),2.37(d,J=12.9Hz,1H),2.30–2.23(m,1H),1.78–1.09(m,28H),1.04(s,3H),0.92(s,4H),0.76(s,3H).
LC-MS:544.8(M+1)
化合物I-15
1H NMR(400MHz,CDCl3)δ6.90(s,1H),4.81(dt,J=2.2,1.2Hz,1H),4.75(dt,J=2.2,1.1Hz,1H),3.35–3.16(m,2H),2.86(qd,J=12.5,3.5Hz,3H),2.55(d,J=13.0Hz,1H),2.37–2.25(m,2H),2.16(ddd,J=12.4,10.8,4.2Hz,1H),1.84–1.64(m,7H),1.63–1.27(m,24H),1.26–1.12(m,4H),1.10–0.91(m,5H),0.78(d,J=6.3Hz,6H).
LC-MS:561.9(M+1)
化合物I-16
1H NMR(400MHz,CDCl3)δ6.91(s,1H),4.85(dt,J=2.1,1.2Hz,1H),4.72(dt,J=2.1,1.1Hz,1H),2.94(dt,J=12.4,5.3Hz,1H),2.79(dt,J=12.5,5.3Hz,1H),2.49(d,J=13.1Hz,1H),2.31(d,J=13.1Hz,1H),2.28–2.17(m,1H),1.88–0.99(m,45H),0.77(s,3H),0.74(s,3H).
LC-MS:560.9(M+1)
化合物I-17
1H NMR(400MHz,CDCl3)δ6.88(s,1H),4.86(dt,J=2.4,1.2Hz,1H),4.70(dt,J=2.4,1.1Hz,1H),3.30(td,J=12.0,2.6Hz,1H),3.17–3.04(m,2H),2.97(td,J=11.9,2.6Hz,1H),2.86(td,J=12.0,2.9Hz,1H),2.49(d,J=12.9Hz,1H),2.33(s,1H),2.24(qt,J=6.7,0.9Hz,1H),2.13(td,J=12.1,2.9Hz,1H),1.96–1.83(m,4H),1.82–1.02(m,34H),0.79(d,J=8.8Hz,6H).
LC-MS:547.9(M+1)
化合物I-18
1H NMR(400MHz,CDCl3)δ6.84(s,1H),4.80(dt,J=2.1,1.1Hz,1H),4.72(dt,J=2.1,1.1Hz,1H),4.08–3.87(m,4H),3.31(td,J=12.1,3.4Hz,1H),3.20(td,J=12.1,3.1Hz,1H),2.99–2.84(m,3H),2.55(d,J=13.0Hz,1H),2.34–2.24(m,2H),2.17(td,J=12.1,3.0Hz,1H),2.10–1.94(m,2H),1.88–1.77(m,1H),1.71–1.63(m,4H),1.62–1.09(m,22H),1.07(s,4H),0.96(q,J=7.0Hz,1H),0.79(s,3H),0.77(s,3H).
LC-MS:563.9(M+1)
化合物I-19
1H NMR(400MHz,CDCl3)δ6.89(s,1H),4.83(dt,J=2.2,1.1Hz,1H),4.76(dt,J=2.2,1.1Hz,1H),3.31–3.18(m,2H),2.94–2.84(m,1H),2.79(ddd,J=12.3,2.9,1.6Hz,1H),2.70–2.49(m,5H),2.34–2.21(m,5H),2.20–1.95(m,4H),1.86–1.63(m,5H),1.63–0.92(m,27H),0.78(d,J=6.6Hz,6H).
LC-MS:576.9(M+1)
化合物I-20
1H NMR(400MHz,CDCl3)δ6.86(s,1H),4.81(dt,J=2.2,1.1Hz,1H),4.72(dt,J=2.2,1.1Hz,1H),3.51(td,J=12.3,3.1Hz,1H),3.40–3.17(m,5H),3.02–2.82(m,3H),2.73(td,J=12.2,3.0Hz,1H),2.52(ddd,J=12.6,5.1,2.9Hz,2H),2.36(d,J=12.9Hz,1H),2.33–2.16(m,2H),1.73–1.27(m,25H),1.27–1.17(m,3H),1.11(dt,J=12.5,6.9Hz,1H),1.04(s,3H),0.91(s,3H),0.75(s,3H).
LC-MS:611.9(M+1)
化合物I-21
于100mL圆底烧瓶中,加入100mg(0.22mmol)化合物6,溶于50mL二氯甲烷中,加入93μL(0.66mmol)TEA,催化量DMAP,41mg(0.44mmol)氨基乙酰氯,室温过夜。减压蒸干,粗品用硅胶柱层析纯化(二氯甲烷:甲醇=20:1),得棕红色固体,收率43%。
1H NMR(400MHz,CDCl3)δ6.79(s,1H),4.72(dt,J=2.2,1.1Hz,1H),4.49(dt,J=2.1,1.1Hz,1H),4.39(d,J=12.4Hz,1H),3.40(d,J=12.3Hz,1H),2.61(d,J=12.9Hz,1H),2.30(d,J=12.9Hz,1H),2.20–2.01(m,2H),1.85–1.64(m,8H),1.62–1.46(m,3H),1.44–1.09(m,16H),1.08–0.95(m,4H),0.86(s,3H),0.78(s,3H).
LC-MS:507.8(M+1)
化合物I-22
1H NMR(400MHz,CDCl3)δ6.85(s,1H),4.86(dt,J=2.2,1.1Hz,1H),4.71(dt,J=2.4,1.2Hz,1H),3.90(t,J=6.8Hz,1H),2.89(dt,J=9.5,6.9Hz,1H),2.62(dt,J=9.5,6.9Hz,1H),2.54(d,J=13.0Hz,1H),2.38(d,J=13.0Hz,1H),2.30(ddd,J=8.1,7.0,5.9Hz,2H),2.23–2.12(m,1H),1.93–1.76(m,4H),1.75–1.60(m,5H),1.60–1.09(m,21H),1.04(s,3H),0.94(d,J=16.0Hz,4H),0.73(s,3H).
LC-MS:547.8(M+1)
化合物I-23
1H NMR(400MHz,CDCl3)δ6.88(s,1H),4.82(dt,J=2.4,1.1Hz,1H),4.74(dt,J=2.1,1.2Hz,1H),2.99(ddt,J=19.5,12.5,7.2Hz,2H),2.54–2.44(m,2H),2.40–2.28(m,4H),2.25–2.07(m,3H),2.01–1.55(m,14H),1.53–1.27(m,15H),1.27–0.99(m,7H),0.89(s,3H),0.76(s,3H).
LC-MS:575.9(M+1)
化合物I-24
1H NMR(400MHz,CDCl3)δ6.91(s,1H),4.81(dt,J=2.1,1.1Hz,1H),4.74(dt,J=2.1,1.1Hz,1H),3.64(d,J=12.4Hz,1H),3.23–3.04(m,3H),2.51(d,J=13.0Hz,1H),2.34(d,J=13.0Hz,1H),2.18(qt,J=7.1,1.2Hz,1H),2.04–1.84(m,5H),1.83–1.67(m,9H),1.66–1.52(m,3H),1.51–1.27(m,14H),1.27–1.11(m,3H),1.05(d,J=19.9Hz,4H),0.82(s,3H),0.71(s,3H).
LC-MS:561.9(M+1)
化合物I-25
1H NMR(400MHz,CDCl3)δ6.85(s,1H),4.81(dt,J=2.1,1.1Hz,1H),4.73(dt,J=2.4,1.1Hz,1H),4.26(s,2H),3.85(p,J=7.0Hz,1H),2.51(d,J=13.0Hz,1H),2.34(d,J=13.0Hz,1H),2.17(qt,J=7.1,1.2Hz,1H),2.05–1.92(m,1H),1.87–1.68(m,11H),1.67–1.27(m,21H),1.26–1.12(m,2H),1.09–0.99(m,5H),0.92(s,3H).0.77(s,3H).
LC-MS:576.9(M+1)
化合物I-26
1H NMR(400MHz,CDCl3)δ6.89(s,1H),4.86(dt,J=2.2,1.1Hz,1H),4.74(dt,J=2.3,1.1Hz,1H),4.05–3.83(m,2H),3.13(dt,J=12.5,2.4Hz,1H),3.01(tdd,J=12.3,8.4,2.6Hz,2H),2.84(td,J=12.3,2.7Hz,1H),2.59–2.45(m,3H),2.36(d,J=13.0Hz,1H),2.28–2.21(m,1H),2.20–2.09(m,2H),1.98(dt,J=12.5,2.4Hz,1H),1.83–1.76(m,2H),1.75–1.18(m,24H),1.17–1.02(m,5H),0.87(s,3H),0.75(s,3H).
LC-MS:593.9(M+1)
化合物I-27
1H NMR(400MHz,CDCl3)δ6.93(s,1H),4.94(dt,J=2.2,1.1Hz,1H),4.78–4.68(m,1H),4.20–4.12(m,2H),4.11–3.99(m,2H),3.65(d,J=12.3Hz,1H),3.18(d,J=12.3Hz,1H),3.03(dt,J=12.4,1.9Hz,1H),2.94–2.76(m,2H),2.52(d,J=13.0Hz,1H),2.37–2.25(m,2H),2.06–2.00(m,1H),1.96–1.83(m,2H),1.81–1.66(m,6H),1.64–1.09(m,21H),1.01(s,3H),0.77(s,3H),0.67(s,3H).
LC-MS:577.9(M+1)
化合物I-28
1H NMR(400MHz,CDCl3)δ6.91(s,1H),4.94(dt,J=2.5,1.2Hz,1H),4.74(dt,J=2.2,1.0Hz,1H),3.60(d,J=12.3Hz,1H),3.14(d,J=12.5Hz,1H),2.98–2.59(m,5H),2.52(d,J=13.0Hz,1H),2.40–2.02(m,8H),1.94–1.08(m,29H),1.04(s,3H),0.81(s,3H),0.70(s,3H).
LC-MS:590.9(M+1)
化合物I-29
1H NMR(400MHz,CDCl3)δ6.94(s,1H),4.91(dt,J=2.2,1.2Hz,1H),4.72(dt,J=2.3,1.2Hz,1H),3.61(d,J=12.5Hz,1H),3.14(d,J=12.5Hz,1H),3.02–2.76(m,4H),2.71–2.63(m,2H),2.52(d,J=12.9Hz,1H),2.39–2.26(m,2H),2.26–2.01(m,4H),1.94–1.67(m,8H),1.65–1.13(m,20H),1.07(t,J=8.0Hz,4H),0.99(s,3H),0.77(s,3H),0.66(s,3H).
LC-MS:604.9(M+1)
化合物I-30
1H NMR(400MHz,CDCl3)δ6.91(s,1H),4.86(dt,J=2.2,1.1Hz,1H),4.69(dt,J=2.4,1.2Hz,1H),4.46(t,J=6.9Hz,1H),2.53(d,J=13.0Hz,1H),2.40–2.23(m,4H),2.10–2.04(m,2H),2.04–1.92(m,1H),1.82–1.27(m,23H),1.25–1.07(m,4H),1.02(s,3H),0.85(d,J=23.0Hz,4H),0.76(s,3H).
LC-MS:561.8(M+1)
化合物I-31
于100mL圆底烧瓶中,加入100mg(0.21mmol)化合物5,溶于50mL四氢呋喃中,加入69μL(0.42mmol)DIPEA,30mg(0.32mmol)甲基胺(30%醇溶液),室温过夜。加入150mL水中,50mL乙酸乙酯萃取三次,合并有机相,饱和食盐水洗,无水硫酸钠干燥。减压旋干,硅胶柱层析(石油醚:乙酸乙酯=1:1)。得黄色固体,收率30%
1H NMR(500MHz,CDCl3)δ6.89(s,1H),4.85(s,1H),4.76(dd,J=2.0,1.1Hz,1H),2.85(s,3H),2.56(d,J=13.0Hz,1H),2.38(d,J=13.0Hz,1H),2.30(qt,J=7.0,1.0Hz,1H),1.93(dt,J=12.8,7.0Hz,1H),1.84–1.14(m,27H),1.04(s,3H),0.94(d,J=21.2Hz,4H),0.79(s,3H).
LC-MS:507.8(M+1)
化合物I-32
以化合物5为原料,参照化合物I-31的合成过程,将甲基胺替换成环丙胺。得浅黄色固体,收率为28%。
1H NMR(400MHz,CDCl3)δ6.79(s,1H),4.77–4.66(m,2H),2.54(d,J=12.9Hz,2H),2.47–2.25(m,4H),1.90–1.15(m,25H),1.13–0.97(m,5H),0.89(s,3H),0.78(s,3H),0.67–0.57(m,2H),0.54–0.45(m,2H).
LC-MS:533.8(M+1)
化合物I-33
以化合物5为原料,参照化合物I-31的合成过程,将甲基胺替换成环丙基甲基胺。得浅黄色固体,收率为41%。
1H NMR(500MHz,CDCl3)δ6.89(s,1H),4.87(dt,J=2.3,1.2Hz,1H),4.74(dd,J=2.0,1.1Hz,1H),3.10(d,J=7.1Hz,2H),2.59–2.45(m,2H),2.41–2.28(m,2H),1.83–1.27(m,23H),1.27–1.11(m,3H),1.09–0.95(m,5H),0.94–0.84(m,4H),0.79(s,3H),0.50–0.40(m,2H),0.27–0.13(m,2H).
LC-MS:547.8(M+1)
化合物I-34
1H NMR(400MHz,CDCl3)δ6.84(s,1H),4.83(dt,J=2.1,1.1Hz,1H),4.66(dt,J=2.3,1.1Hz,1H),4.28–4.05(m,1H),3.27–3.06(m,2H),2.92–2.69(m,2H),2.56(dd,J=14.5,12.2Hz,2H),2.44–2.17(m,3H),2.09–1.92(m,2H),1.88–0.88(m,40H),0.77(s,3H).
LC-MS:592.9(M+1)
化合物I-35
1H NMR(400MHz,CDCl3)δ6.90(s,1H),4.81(dt,J=2.1,1.1Hz,1H),4.77(dt,J=2.2,1.1Hz,1H),4.65(ddd,J=12.5,3.1,1.6Hz,1H),3.79–3.61(m,2H),3.28(td,J=12.3,2.8Hz,1H),2.87–2.71(m,2H),2.52(d,J=13.0Hz,2H),2.34(d,J=16.3Hz,4H),2.26–2.12(m,2H),1.97(dtd,J=12.6,6.9,1.1Hz,2H),1.88–1.00(m,30H),0.87(s,3H),0.81(s,3H).
LC-MS:576.9(M+1)
化合物I-36
1H NMR(400MHz,CDCl3)δ6.81(s,1H),4.78(dt,J=2.2,1.2Hz,1H),4.72–4.63(m,3H),3.42(dddd,J=12.3,6.7,5.0,2.6Hz,2H),3.26(dt,J=6.8,1.0Hz,1H),3.05(dt,J=12.5,2.4Hz,2H),2.88(td,J=12.3,2.7Hz,2H),2.55(d,J=13.0Hz,1H),2.29(d,J=13.0Hz,1H),2.18–1.26(m,24H),1.26–1.02(m,8H),0.77(s,3H),0.65(s,3H).
LC-MS:579.9(M+1)
化合物I-37
1H NMR(400MHz,CDCl3)δ6.93(s,1H),4.86(dt,J=2.2,1.1Hz,1H),4.75(dt,J=2.2,1.1Hz,1H),4.57–4.34(m,2H),4.17(ddd,J=11.4,3.0,1.2Hz,2H),3.90(td,J=11.6,2.3Hz,2H),3.41–3.28(m,2H),2.68–2.47(m,2H),2.44–2.25(m,2H),1.93–1.09(m,26H),1.08–0.87(m,8H),0.78(s,3H).
LC-MS:563.8(M+1)
化合物I-38
1H NMR(400MHz,CDCl3)δ6.97(s,1H),5.28(td,J=12.4,2.8Hz,1H),4.78(dt,J=2.2,1.1Hz,1H),4.71(dt,J=2.3,1.2Hz,1H),4.49(dt,J=12.6,2.2Hz,1H),4.22(ddd,J=12.4,9.6,5.2Hz,1H),3.81–3.47(m,2H),3.22(dt,J=7.1,1.1Hz,1H),3.07–2.80(m,3H),2.55(d,J=12.9Hz,1H),2.29(d,J=13.0Hz,1H),2.06–0.98(m,32H),0.75(s,3H),0.64(s,3H).
LC-MS:611.9(M+1)
化合物I-39
1H NMR(400MHz,CDCl3)δ8.42(d,J=5.1Hz,2H),7.93(d,J=5.1Hz,2H),6.84(s,1H),4.88(dt,J=2.3,1.2Hz,1H),4.79(dt,J=2.3,1.2Hz,1H),2.64–2.50(m,2H),2.45–2.27(m,2H),1.94–1.12(m,26H),1.09–0.95(m,5H),0.91(s,3H),0.80(s,3H).
LC-MS:570.8(M+1)
化合物I-40
1H NMR(400MHz,CDCl3)δ6.92(s,1H),4.87(dt,J=2.4,1.2Hz,1H),4.74(dt,J=2.2,1.0Hz,1H),4.52(dt,J=12.5,7.1Hz,1H),4.29(dt,J=12.6,7.2Hz,1H),4.03–3.88(m,1H),3.11(ddt,J=57.9,12.4,7.1Hz,2H),2.61–2.50(m,2H),2.42–2.10(m,4H),2.01–1.88(m,1H),1.84–1.27(m,33H),1.27–1.11(m,3H),1.04(s,5H),0.90(s,3H),0.79(s,3H).
LC-MS:677.0(M+1)
化合物I-41
1H NMR(400MHz,CDCl3)δ6.88(s,1H),4.90(dt,J=2.2,1.1Hz,1H),4.70(dt,J=2.1,1.1Hz,1H),4.37(p,J=6.9Hz,1H),4.05(dd,J=12.3,7.0Hz,1H),3.97–3.71(m,2H),3.40(dd,J=12.3,7.0Hz,1H),2.94(qt,J=7.2,1.2Hz,1H),2.60–2.28(m,3H),2.19–0.98(m,35H),0.88(s,3H),0.82(s,3H).
LC-MS:577.9(M+1)
化合物I-42
1H NMR(400MHz,CDCl3)δ6.95(s,1H),4.91(dt,J=2.3,1.1Hz,1H),4.69(dt,J=2.3,1.0Hz,1H),4.14(td,J=12.1,2.5Hz,1H),3.17(td,J=9.3,7.8Hz,1H),3.10–2.76(m,4H),2.60–2.29(m,4H),2.17(td,J=9.2,1.2Hz,1H),2.02–1.06(m,33H),1.02(s,3H),0.83(s,3H),0.75(s,3H).
LC-MS:590.9(M+1)
化合物I-43
1H NMR(400MHz,CDCl3)δ6.99(s,1H),4.90(dt,J=2.1,1.1Hz,1H),4.73–4.64(m,1H),4.21–3.93(m,5H),3.30(td,J=12.0,4.1Hz,1H),3.12–2.82(m,3H),2.57–2.29(m,5H),2.19(dd,J=4.0,1.2Hz,1H),2.03(d,J=12.6Hz,1H),1.94–1.07(m,26H),1.05(s,5H),0.91(s,3H),0.81(s,3H).
LC-MS:606.9(M+1)
化合物I-44
1H NMR(400MHz,CDCl3)δ6.89(s,1H),4.81(dt,J=2.1,1.1Hz,1H),4.70(dt,J=2.2,1.1Hz,1H),4.11(ddd,J=12.4,11.3,3.4Hz,2H),3.67(dt,J=12.5,2.2Hz,1H),3.39–3.08(m,3H),2.99–2.80(m,2H),2.80–2.32(m,6H),2.01–1.87(m,2H),1.85–1.28(m,24H),1.24–1.01(m,7H),0.85(s,3H),0.78(s,3H).
LC-MS:655.9(M+1)
化合物I-45
1H NMR(400MHz,CDCl3)δ6.98(s,1H),4.90(dt,J=2.2,1.1Hz,1H),4.69(dt,J=2.1,1.1Hz,1H),4.03(td,J=11.8,3.4Hz,1H),3.42–3.16(m,2H),3.01–2.91(m,1H),2.85(ddd,J=12.5,3.2,1.6Hz,1H),2.71–2.40(m,5H),2.37–2.15(m,7H),2.13–2.01(m,2H),1.91–1.05(m,28H),1.02(s,3H),0.88(s,3H),0.82(s,3H).
LC-MS:619.9(M+1)
化合物I-46
以化合物5为原料,参照化合物I-31的合成过程,将甲基胺替换成乙醇。得浅黄色固体,收率为37%
1H NMR(400MHz,CDCl3)δ6.89(s,1H),5.22(dq,J=12.4,8.0Hz,1H),4.81(dt,J=2.1,1.1Hz,1H),4.70(dt,J=2.4,1.2Hz,1H),3.76(dq,J=12.3,8.0Hz,1H),2.54(d,J=13.0Hz,1H),2.40–2.19(m,2H),1.94(ddt,J=12.9,9.2,7.1Hz,2H),1.79(t,J=7.0Hz,1H),1.74–1.56(m,5H),1.53–1.26(m,16H),1.26–1.03(m,11H),1.01(s,3H),0.76(s,3H).
LC-MS:522.8(M+1)
化合物I-47
1H NMR(400MHz,CDCl3)δ6.83(s,1H),4.87(dt,J=2.3,1.1Hz,1H),4.73(dt,J=2.3,1.2Hz,1H),3.84(ddt,J=17.6,11.4,7.0Hz,2H),3.48(ddt,J=24.5,11.6,7.1Hz,2H),3.10(d,J=7.0Hz,2H),2.64–2.49(m,2H),2.43–2.15(m,3H),1.83–1.28(m,25H),1.27–1.12(m,5H),1.10–0.94(m,5H),0.90(s,3H),0.79(s,3H).
LC-MS:591.9(M+1)
化合物I-48
1H NMR(400MHz,CDCl3)δ6.98(s,1H),4.90(dt,J=2.2,1.1Hz,1H),4.77(dt,J=2.1,1.1Hz,1H),4.37(p,J=6.9Hz,1H),4.05(dd,J=12.3,7.0Hz,1H),3.98–3.86(m,1H),3.78(dt,J=8.0,7.0Hz,1H),3.40(dd,J=12.3,7.0Hz,1H),2.94(qt,J=7.2,1.2Hz,1H),2.52(d,J=13.1Hz,1H),2.49–2.39(m,1H),2.34(d,J=13.0Hz,1H),2.15–1.99(m,2H),1.91–1.06(m,30H),1.03(s,3H),0.88(s,3H),0.81(s,3H).
LC-MS:577.9(M+1)
化合物I-49
1H NMR(400MHz,CDCl3)δ8.79(dd,J=5.0,1.0Hz,2H),7.39(t,J=5.0Hz,1H),6.92(s,1H),5.04(d,J=12.5Hz,1H),4.97–4.83(m,2H),4.74(dt,J=2.3,1.1Hz,1H),2.67–2.49(m,2H),2.46–2.25(m,2H),1.85–1.57(m,10H),1.56–1.28(m,13H),1.25–1.11(m,3H),1.08(d,J=15.6Hz,5H),0.90(s,3H),0.80(s,3H).
LC-MS:585.9(M+1)
化合物I-50
1H NMR(400MHz,CDCl3)δ6.83(s,1H),4.86(dt,J=2.2,1.0Hz,1H),4.77(dt,J=2.2,1.0Hz,1H),4.66–4.51(m,2H),4.28(dt,J=8.1,7.0Hz,1H),2.76(dq,J=13.1,7.1Hz,1H),2.60–2.44(m,2H),2.39–2.21(m,4H),1.88–1.74(m,3H),1.70(d,J=1.3Hz,3H),1.66–1.27(m,15H),1.25–1.04(m,7H),1.04–0.91(m,2H),0.75(d,J=12.6Hz,6H).
LC-MS:577.8(M+1)
化合物I-51
以化合物I-11为原料,参考化合物I-52合成过程,得棕色固体,收率91%。
1H NMR(400MHz,CDCl3)δ6.43(s,1H),3.17(dt,J=12.5,7.1Hz,2H),2.52–2.31(m,4H),2.30–2.19(m,5H),1.99–0.87(m,36H),0.83–0.71(m,12H).
LC-MS:563.9(M+1)
化合物I-52
以化合物I-7为原料,参考化合物I-52合成过程,得黄色固体,收率87%。
1H NMR(400MHz,CDCl3)δ7.78(s,1H),4.22(s,1H),3.89–3.60(m,2H),2.78–2.52(m,2H),2.09–1.91(m,4H),1.70–1.09(m,33H),0.89(tt,J=41.4,7.1Hz,12H).
LC-MS:536.8(M+1)
化合物I-53
1H NMR(400MHz,DMSO-d6)δ12.10(s,1H),δ7.05(d,J=4.5Hz,1H),4.80–4.69(m,1H),4.61(s,1H),2.59(ddd,J=24.9,12.1,5.0Hz,3H),2.44–1.88(m,6H),1.79–1.04(m,32H),0.98(d,J=5.8Hz,4H),0.92–0.83(m,1H),0.79(s,6H).
LC-MS:533.8(M+1)
化合物I-54
1H NMR(400MHz,CDCl3)δ7.12(s,1H),4.79(s,1H),4.76(s,1H),2.74–2.59(m,5H),2.42–2.11(m,5H),1.94–0.82(m,39H),0.74(d,J=4.7Hz,6H).
LC-MS:561.9(M+1)
化合物I-55
所得固体溶于30mL二氯甲烷,加入6mL三氟乙酸,室温过夜,减压蒸干,30mL饱和碳酸氢钠水溶液洗涤,加入100mL水,50mL二氯甲烷萃取三次,饱和食盐水洗,无水硫酸钠干燥,柱层析纯化(二氯甲烷:甲醇=10:1)。得黄色固体,收率97%。
1H NMR(400MHz,CDCl3)δ7.77–7.58(m,1H),4.77(s,1H),4.62(d,J=10.2Hz,1H),2.83–2.67(m,4H),2.60(d,J=14.9Hz,2H),2.44–1.88(m,3H),1.82–1.07(m,34H),1.06–0.64(m,9H).
LC-MS:547.9(M+1)
化合物I-56
1H NMR(400MHz,CDCl3)δ7.09(s,1H),4.79(dt,1H),4.67(dt,1H),3.21–3.17(m,2H),3.02–2.57(m,8H),2.36–2.09(m,3H),1.92–0.93(m,34H),0.82(d,J=6.3Hz,6H).
LC-MS:562.9(M+1)
生物活性测试实验
一.通过巨噬细胞的非特异性毒性作用及体外诱导的破骨细胞分化体系评价化合物对破骨细胞形成的抑制活性,方法如下:
1.主要使用实验材料及仪器
(1)细胞系:小鼠单核巨噬白血病细胞RAW 264.7(美国ATCC公司);
(2)RANKL(Sigma公司);M-CSF(Peprotech公司);MTT(Sigma公司);DMEM培养液(Gibco公司);FBS胎牛血清,(Hyclone公司);抗酒石酸酸性磷酸酶检测试剂盒(Sigma公司)
(3)Spectra 190全波长酶标仪(Molecular Devices公司);细胞培养箱(Thermo公司)
2.实验方法
(1)毒性测试:
RAW264.7细胞1500个/孔接种于96孔板,同时加入不同浓度化合物,另设相应的溶媒对照(细胞对照)及培养液本底对照(空白对照)。37℃,5%CO2培养箱中培养48小时。结束培养前4小时加入MTT溶液。至培养结束,吸弃上清,每孔加入DMSO溶解紫色结晶,于酶标仪570nM处测定OD值。
(2)破骨细胞分化抑制活性测试:
RAW264.7细胞1500个/孔接种于96孔板,加入刺激剂RANKL(终浓度30ng/ml)、M-CSF(终浓度40ng/ml)及不同浓度化合物,另设相应的无药物阳性对照孔及无刺激剂的阴性对照,37℃,5%CO2培养箱中培养96小时。培养终点进行抗酒石酸酸性磷酸酶(Tartrateresistant acid phosphatase,TRAP)染色,显微镜下计数阳性染色细胞数。
结果评定:
化合物对RAW264.7细胞的非特异性毒性作用采用待测样品OD值除以细胞对照孔OD值,标记为细胞存活率(%)。破骨细胞分化程度采用阳性对照样品阳性染色细胞数减去被检测样品阳性染色细胞数,然后除以阳性染色细胞数,标记为破骨分化抑制率(%)。CC50和IC50值使用GraphPad Prism 6软件拟合。
结果见表1.
表1.部分化合物对RAW264.7细胞毒性和对破骨细胞分化的抑制活性
*选择指数SI=CC50/IC50
上述结果表明,本发明制备的多数化合物相对于吡唑并白桦脂酸具有较低的毒性,且对破骨细胞分化具有较好的抑制作用,尤其是化合物I-6、I-7、I-11、I-12、I-30、I-50对RANKL/M-CSF诱导的破骨细胞分化具有显著抑制作用,抑制活性和选择指数SI高于阳性参照物双醋瑞因和起始衍生物吡唑并白桦脂酸。
二.通过细菌脂多糖(LPS)诱导的巨噬细胞炎症因子分泌体系评价化合物的抗炎活性,方法如下:
RAW264.7细胞6×105个/孔接种于24孔板,加入LPS(终浓度5μg/ml)及不同浓度化合物,另设相应的无药物阳性对照孔及无刺激及的阴性对照,37℃,5%CO2培养箱中培养48小时。培养终点收集细胞培养上清,检测TNF-α、IL-1β、IL-6的含量。
结果见表2及图1
表2.部分制备化合物对LPS诱导RAW264.7分泌炎症因子的抑制活性
如图1所示,化合物I-7、I-11、I-12、I-30、I-50及双醋瑞因在无细胞毒性浓度范围内对于LPS诱导的RAW264.7细胞分泌IL-1β均有较高的抑制活性,其中化合物I-11,I-12及双醋瑞因对于IL-1β的抑制作用具有浓度依赖性。
以上结果表明,本发明制备的化合物I-7、I-11、I-12、I-30、I-50对巨噬细胞分泌IL-1β具有显著抑制作用。综合化合物对于破骨细胞分化的抑制活性,化合物I-11的抗炎作用和分化抑制作用具有更为理想的活性窗口。
二.通过碘乙酸钠诱导的大鼠关节炎模型评价化合物对骨关节炎的治疗作用,方法如下:
1.主要使用实验材料及仪器
(1)动物
SPF级SD大鼠,雄性,体重280-300g,由北京华阜康生物科技股份有限公司提供,合格证编号:No.1103222011009688。饲养于中国科学院上海药物研究所SPF级动物房。
(2)主要实验药物及试剂
碘乙酸钠(Sigma公司,CAS:305-53-3);0.9%氯化钠注射液(生理盐水,华裕(无锡)制药有限公司);地塞米松磷酸钠注射液(规格5mg/mL,1mL/支,辰欣药业股份有限公司);羟丙基甲基纤维素(HPMC,Sigma公司);聚氧乙烯氢化蓖麻油(ELP,Kolliphor);其余试剂均为国产分析纯试剂。
(3)主要仪器
双足平衡测试仪(美国IITC,600MR型);冯·弗雷纤维丝(美国North CoastNC12775-99型)
2.实验方法
(1)碘乙酸钠(MIA)诱导大鼠关节炎模型构建
将SD大鼠分为2组,正常组和模型组。碘乙酸钠以无菌水配制成5w/v%浓度,模型组大鼠左膝单次关节腔内注射20μl/只。
(2)化合物关节腔注射液配制:化合物I11 4mg溶于由75μL 0.2%HPMC加入25μLELP的混合溶剂中,得到40mg/mL储备液;取20μL储备液加入180μL生理盐水中,获得4mg/mL溶液(高剂量组)(溶剂体系:0.2%HPMC:ELP:生理盐水=0.75:0.25:9);4mg/mL溶液以生理盐水稀释2倍,获得2mg/mL溶液(低剂量组)。
(3)化合物对碘乙酸钠诱导大鼠关节炎的治疗作用
1)模型组大鼠造模后第3天进行左侧足痛阈及单侧负重能力(冯·弗雷纤维丝及双足平衡测试)检测,并根据检测值将模型大鼠平均分成4组:模型组、阳性参照物地塞米松治疗组,I-11高剂量治疗组及I-11低剂量治疗组。各组大鼠每周关节腔注射1次,25μL/次/只:溶剂对照组(0.2%HPMC:ELP:生理盐水=0.75:0.25:9),地塞米松治疗组(地塞米松磷酸钠注射液,125μg/只),I-11高剂量治疗组(4mg/mL溶液,100μg/只),I11低剂量治疗组(2mg/mL溶液,50μg/只)。从分组当天起,共治疗2周,关节腔注射2次。
2)小鼠关节炎疼痛程度评价:
造模第0天、第3天、第7天、第10天及第14天,用冯·弗雷纤维丝检测各组大鼠50%缩足疼痛阈值(图2);
造模第0天、第3天、第7天、第10天及第14天,用双足平衡仪检测各组大鼠单测负重/承重(图3);
(注:各组与模型组比较#=P<0.05,##=P<0.01,###=P<0.001)。
(4)实验结果
如图2及图3所示,采用I-11关节腔注射MIA诱导的关节炎大鼠,1周1次治疗能够显著缓解模型动物关节疼痛程度,其中100μg高剂量的I-11治疗的作用效果与125μg的地塞米松阳性参照物相当。
以上结果表明,化合物I-11关节腔注射对于骨关节炎主要症状之一关节炎症疼痛具有显著的改善作用,化合物I-11对于骨关节炎疾病模型的治疗活性优于阳性参照物,具有开发为骨关节炎及炎症性关节病的治疗药物的潜力。
在本发明提及的所有文献都在本申请中应用作为参考,就如同每一篇文献被单独引用作为参考那样。此外应理解,在阅读了本发明的上述讲述内容之后,本领域技术人员可以对本发明作各种改动或修改,这些等价形式同样落于本申请所附权利要求书所限定的范围。
Claims (8)
1.一种如通式I中所述的基于吡唑并白桦脂酸的化合物,其异构体及药学上可接受的盐:
其中,
R1选自以下组:异丙基、异丙烯基;
R2选自以下组:氢、取代或未取代的C1-C4直链、支链或环状烷基;
R3选自以下组:氢、取代或未取代的C1-C4直链或支链烷基、取代或未取代的C3-C6的环烷基、取代或未取代的C1-C4直链或支链烷氧基、取代或未取代的C3-C6的环烷基氧基、取代或未取代的C1-C4直链或支链烷基氨基、取代或未取代的含氧或氮或硫的饱和五元或六元杂环基或其氧化物、取代或未取代的含氧或氮或硫的不饱和五元或六元杂环基;
X不存在,或者选自以下组:亚甲基、羰基、-CONH-;
n为0-4的整数;
其中,所述的取代指被选自以下组的取代基取代:卤素、C1-C8直链或支链烷基、C1-C8直链或支链烷基氨基、C1-C8直链或支链烷氧羰基、羟基、-NH2、C1-C8直链或支链烷氧基、氧代基团。
6.一种药物组合物,所述的药物组合物含有治疗有效量的选自根据权利要求1-3任一项中所述的化合物、其异构体及药学上可接受的盐中的一种或多种,以及任选地,药学上可接受的辅料。
7.根据权利要求1-3任一项所述的基于吡唑并白桦脂酸的化合物、其异构体及药学上可接受的盐或根据权利要求6所述的药物组合物在制备治疗与破骨细胞活性抑制、免疫活性抑制、炎症因子活性抑制相关的疾病的药物中的用途。
8.根据权利要求7所述的用途,其中,所述与破骨细胞活性抑制、免疫活性抑制、炎症因子活性抑制相关的疾病包括骨质疏松症、骨关节炎、牙周炎、牙齿脱落、Paget’s骨病、佝偻病、骨巨细胞瘤、骨髓瘤骨病以及癌症骨转移造成的骨破坏。
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CN104974215A (zh) * | 2014-04-02 | 2015-10-14 | 华东师范大学 | 白桦脂酸-氨基酸衍生物及其制备方法和应用 |
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RU2021101393A3 (zh) * | 2022-07-26 | |||
US20020037889A1 (en) * | 2000-01-20 | 2002-03-28 | Duggan Mark E. | Alpha V integrin receptor antagonists |
US20050014722A1 (en) * | 2001-12-13 | 2005-01-20 | Gil-Ja Jhon | Process for preparing n-acylated lysophosphatidylcholine and pharmaceutical composition for treatment of metabolic bone disease comprising said compounds |
CN104974215A (zh) * | 2014-04-02 | 2015-10-14 | 华东师范大学 | 白桦脂酸-氨基酸衍生物及其制备方法和应用 |
WO2018041260A1 (zh) * | 2016-09-05 | 2018-03-08 | 中国科学院上海药物研究所 | 一类溴结构域识别蛋白抑制剂及其制备方法和用途 |
WO2018069086A1 (en) * | 2016-10-13 | 2018-04-19 | Vivacell Biotechnology España S.L. | Hydroxamate triterpenoid derivatives |
KR20190015942A (ko) * | 2017-08-07 | 2019-02-15 | 주식회사 코팜 | 골 질환 예방 또는 치료용 화합물 및 이의 용도 |
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