2-酰基五员不饱和杂环化合物的合成及其抗骨质疏松与抗糖尿病活性研究
摘要
依据分子生物学研究的最新进展与成果,本研究所建立了以骨形态发生蛋白-2(BMP-2)为靶点的新型抗骨质疏松药物体外活性筛选模型及含有pBIND-PPAR_γ-LBD表达质粒的新型抗糖尿病药物的体外活性筛选模型,筛选并发现本课题组合成的2-乙酰苯并噻吩和2-(β-羰基丁酰)呋喃分别具有上调BMP-2和PPAR_γ的活性。在此基础上,本论文进行了2-酰基五员不饱和杂环化合物的合成及其抗骨质疏松与抗糖尿病的活性研究。主要研究内容与进展为:
1、论文共合成化合物31个,其中,新苯并五员不饱和杂环目标化合物20个,其结构均经~1H NMR和MS确证。
2、测定了110个2-酰基五员不饱和杂环化合物的体外BMP-2上调活性,其中,具有明显上调BMP-2活性的化合物77个,化合物G31、G38、G28、Shy-B13、G30、Shy-B15、Shy-A39、G13、G25、G26、Shy-S4、G24、G35、G32、G33和G34的活性是对照药洛伐他汀的5倍以上;先导化合物2-乙酰苯并噻吩(G39)的活性与洛伐他汀相当。
3、测定了2-乙酰苯并噻吩和2-乙酰苯并呋喃(G03)灌胃给药后的大鼠体内抗骨质疏松效果。结果发现,两者均具有明显改善去势大鼠骨质疏松症状的作用,高剂量2-乙酰苯并噻吩的作用与对照药雷洛西芬相当。初步毒性研究结果,小鼠灌胃给予2-乙酰苯并噻吩的LD_(50)>5g/kg,腹腔注射的LD_(50)为1.6g/kg。
4、测定了70多个2-酰基五员不饱和杂环化合物的PPARγ上调活性,其中,上调率大于30%的化合物有Shy-B10、Shy-A12、Shy-A27、Shy-A38、Shy-S2等10个、大于40%的化合物有Shy-B8、Shy-A29等6个。
5、测定了Shy-A12、Shy-A29、Shy-B10、Shy-S17和GHF 5个化合物灌胃给药后的小鼠体内降血糖效果。结果发现,Shy-A12和GHF具有降糖作用,在试验剂量(75mg/kg)下,Shy-A12的降糖效果与对照药罗格列酮相当。
6、初步总结了2-酰基五员不饱和杂环化合物的构效关系,为指导进一步结构优化打下基础。
通过论文研究工作,筛选得到值得进一步深入研究的抗骨质疏松与抗糖尿病活性化合物多个,其中,抗骨质疏松活性化合物G39和抗糖尿病活性化合物Shy-A12具有一定开发应用前景。上述两个化合物的抗骨质疏松作用和抗糖尿病作用均未见有文献报道,也未见该两个化合物具有任何其他生物活性的研究报道。
By using MC3T3E1 cells transfected by recombinant plasmid PMB which targeted bone morphogenetic protein-2 and HepG2 cells transfected by expression vector pBIND-PPAR_γ-LBD,synthetic compounds and natural compounds were screened to evaluate the antiosteoporosis activities and the antidiabetic activities in vitro. 2-acetyl-benzothiophene and 2-(β-carbonylbutyryl)furan were found to have antiosteoporosis activity and antidiabetic activity respectively.
With 2-acetyl-benzothiophene and 2-(β-carbonylbutyryl)furan as lead compounds, structure modification was carried out.31 compounds were synthesized.21 compounds of the synthesized compounds and 70 compounds from compound library were evaluated for their antiosteoporosis activities and antidiabetic activities in vitro and in vivo. In vitro,77compounds were found to have antiosteoporosis activities.The activities of compounds G31,G38,G28,Shy-B13,G30,Shy-B15,Shy-A39,G13,G25,G26,ShyS4, G24,G35,G32,G33 and G34 were more than 5 times of positive control Lovastatin.23 compounds were found to have antidiabetic activies.The up-regulation activities of Shy-B10,Shy-A12,Shy-A27,Shy-A38,Shy-S2 were more than 30%,the up-regulation activities of Shy-B8,Shy-A29 were more than 40%.
In vivo,before the evaluation of antiosteoporosis activities of 2-acetyl-benzothiophene was carried out,the toxicity of the compound was detected.The intragastric administration LD_50 of the compound was more than 5g/kg,the intraperitoneal injection LD_50 was more than 1.6g/kg.Dramatic antiosteoporosis effects of 2-acetyl-benzothiophene and 2-acetyl-benzofuran were found,especially, 2-acetyl-benzothiophene at high dose had comparable effect with positive control Raloxifene.The antidiabetic effects of Shy-A12 and GHF were found.At 75mg/kg dose, Shy-A12 and GHF lowered fast blood glucose in mice,especially,Shy-A12 had comparable effect with positive control Rosiglitazone.
The antiosteoporosis activities of 2-acetyl-benzothiophene and 2-acetyl-benzofuran,and the antidiabetic activity of Shy-A12 were first found.2-acetyl-benzothiophene, 2-acetyl-benzofuran and Shy-A12 had the potential to be the lead compound for antiosteoporosis drugs rsearch and antidiabetic drug research respectively.
1、论文共合成化合物31个,其中,新苯并五员不饱和杂环目标化合物20个,其结构均经~1H NMR和MS确证。
2、测定了110个2-酰基五员不饱和杂环化合物的体外BMP-2上调活性,其中,具有明显上调BMP-2活性的化合物77个,化合物G31、G38、G28、Shy-B13、G30、Shy-B15、Shy-A39、G13、G25、G26、Shy-S4、G24、G35、G32、G33和G34的活性是对照药洛伐他汀的5倍以上;先导化合物2-乙酰苯并噻吩(G39)的活性与洛伐他汀相当。
3、测定了2-乙酰苯并噻吩和2-乙酰苯并呋喃(G03)灌胃给药后的大鼠体内抗骨质疏松效果。结果发现,两者均具有明显改善去势大鼠骨质疏松症状的作用,高剂量2-乙酰苯并噻吩的作用与对照药雷洛西芬相当。初步毒性研究结果,小鼠灌胃给予2-乙酰苯并噻吩的LD_(50)>5g/kg,腹腔注射的LD_(50)为1.6g/kg。
4、测定了70多个2-酰基五员不饱和杂环化合物的PPARγ上调活性,其中,上调率大于30%的化合物有Shy-B10、Shy-A12、Shy-A27、Shy-A38、Shy-S2等10个、大于40%的化合物有Shy-B8、Shy-A29等6个。
5、测定了Shy-A12、Shy-A29、Shy-B10、Shy-S17和GHF 5个化合物灌胃给药后的小鼠体内降血糖效果。结果发现,Shy-A12和GHF具有降糖作用,在试验剂量(75mg/kg)下,Shy-A12的降糖效果与对照药罗格列酮相当。
6、初步总结了2-酰基五员不饱和杂环化合物的构效关系,为指导进一步结构优化打下基础。
通过论文研究工作,筛选得到值得进一步深入研究的抗骨质疏松与抗糖尿病活性化合物多个,其中,抗骨质疏松活性化合物G39和抗糖尿病活性化合物Shy-A12具有一定开发应用前景。上述两个化合物的抗骨质疏松作用和抗糖尿病作用均未见有文献报道,也未见该两个化合物具有任何其他生物活性的研究报道。
By using MC3T3E1 cells transfected by recombinant plasmid PMB which targeted bone morphogenetic protein-2 and HepG2 cells transfected by expression vector pBIND-PPAR_γ-LBD,synthetic compounds and natural compounds were screened to evaluate the antiosteoporosis activities and the antidiabetic activities in vitro. 2-acetyl-benzothiophene and 2-(β-carbonylbutyryl)furan were found to have antiosteoporosis activity and antidiabetic activity respectively.
With 2-acetyl-benzothiophene and 2-(β-carbonylbutyryl)furan as lead compounds, structure modification was carried out.31 compounds were synthesized.21 compounds of the synthesized compounds and 70 compounds from compound library were evaluated for their antiosteoporosis activities and antidiabetic activities in vitro and in vivo. In vitro,77compounds were found to have antiosteoporosis activities.The activities of compounds G31,G38,G28,Shy-B13,G30,Shy-B15,Shy-A39,G13,G25,G26,ShyS4, G24,G35,G32,G33 and G34 were more than 5 times of positive control Lovastatin.23 compounds were found to have antidiabetic activies.The up-regulation activities of Shy-B10,Shy-A12,Shy-A27,Shy-A38,Shy-S2 were more than 30%,the up-regulation activities of Shy-B8,Shy-A29 were more than 40%.
In vivo,before the evaluation of antiosteoporosis activities of 2-acetyl-benzothiophene was carried out,the toxicity of the compound was detected.The intragastric administration LD_50 of the compound was more than 5g/kg,the intraperitoneal injection LD_50 was more than 1.6g/kg.Dramatic antiosteoporosis effects of 2-acetyl-benzothiophene and 2-acetyl-benzofuran were found,especially, 2-acetyl-benzothiophene at high dose had comparable effect with positive control Raloxifene.The antidiabetic effects of Shy-A12 and GHF were found.At 75mg/kg dose, Shy-A12 and GHF lowered fast blood glucose in mice,especially,Shy-A12 had comparable effect with positive control Rosiglitazone.
The antiosteoporosis activities of 2-acetyl-benzothiophene and 2-acetyl-benzofuran,and the antidiabetic activity of Shy-A12 were first found.2-acetyl-benzothiophene, 2-acetyl-benzofuran and Shy-A12 had the potential to be the lead compound for antiosteoporosis drugs rsearch and antidiabetic drug research respectively.
引文
[1]Mundy,G.;Garrett,R.et al.Stimulation of bone formation in vitro and in rodents by statins[J].Science,1999,286:1946-1949.
[2]Roberto S.Nitroflurbiprofen(NicOx)[J].Current Opinion in Investiga tional Drugs,2004,5(5):551-553.
[3]Iwamoto J,Takeda T,Ichimura S.Treatment with vitamin D3 and/or vitamin K2for postmenopausal osteoporosis[J].Keio Journal of Medicine,2003,52(3):147-148.
[4]王葆仁.有机合成化学(下册)[M].北京:科学出版社,1985:880.
[5]王葆仁.有机合成化学(下册)[M].北京:科学出版社,1985:565.
[6]樊能延.有机合成事典[M].北京:北京理工大学出版社,1992:757.
[7]王葆仁.有机合成化学(下册)[M].北京:科学出版社,1985:848.
[8]方朝晖,鲍陶陶,费爱华.骨疏灵对骨质疏松症大鼠骨组织形态的影响[J].中成药,2007,29(4):587-588.
[9]谢 文,冯 坤,陈宝龙.中药对实验性骨质疏松大鼠骨骼影响的形态计量学研究[J].中医正骨,2007,19(3):7-8.
[10]王立英,蔡跃增.骨形态结构的影像学研究方法[J].国外医学临床放射学分册,2007,30(2):115-118.
[11]章明放,张乃鑫,谭郁彬,等.周期性与连续性给予羟乙磷酸钠对去势大鼠骨质疏松症治疗作用的比较[J].中国骨质疏松杂志,1995,1(2):104-107.
[12]J.J.Cees.Tack,Paul Smitts.Troglitazone Decreases the Pro p o rtion of Small,Dense LDL and Increases the Resistance of LDL to Oxidation in Obese S u b j ect [J].E m e r g i n g T r e a t m e n t s a n d T e c h n o l o g i e s,1998,21(5):796-799.
[13]庞建华,张耀苏,杰英.罗格列酮和二甲双肌对高糖高脂诱导的糖尿病脂肪肝大鼠的影响[J].肝脏,2007,112(5):378-381.
[14]常保成,郑少雄.胰岛素增敏剂——罗格列酮在糖尿病中的应用[J].国外医学内分泌分册,2001,2(1):34-36.
[15]王霜,杨正国.降糖新药罗格列酮研究进展[J].临床荟萃,2002,17(7):432-433.
[16]朱波,薛耀明,李晨钟等.罗格列酮对OLETF大鼠胰岛β细胞凋亡水平的影响[J].中国糖尿病杂志,2007,15(1):44-46.
[17]文金生,胡弼,龙光.罗格列酮对果糖饲养大鼠血糖血脂及血管重构的影响[J].中国药理学通报,2004,20(5):571-575.
[18]杨晓峰,李升刚,于得志,等．复方地骨皮冲剂对小鼠血糖血脂及免疫功能的影响[J]．齐鲁医学杂志,2000,15(2)：84-86．
[19]D.M.Ribnickya,A.Pouleva,M.Watfordb.Antihyperglycemic activity of TarralinTM,an ethanolic extract of Artemisia dracunculus L [J].Phytomedicine,2006,13:50-557.
[20]M.M.Ma,J.L.Chen,G.G.Wang,et al.Sphingosine kinase 1 participates in insulin signaling and regulates glucose metabolism and homeostasis in KK/Ay diabetic mice[J].Diabetologia,2007,50:891-900.
[21]Hayato Maeda,Masashi Hosokawa,Tokutake Sashima.Dietary Combination of Fucoxanthin and Fish Oil Attenuates the Weight Gain of White Adipose Tissue and Decreases Blood Glucose in Obese/Diabetic KK-Ay Mice[J].Agric.Food Chem,2007,55:7701-7706.
[22]N.Irwin,P.L.McClean,F.P.M.O'Harte.Early administration of the glucose-dependent insulinotropic polypeptide receptor antagonist (Pro3) GIP prevents the development of diabetes and related metabolic abnormalities associated with genetically inherited obesity in ob/ob mice[J].Diabetologia,2007,50:1532-1540.
[2]Roberto S.Nitroflurbiprofen(NicOx)[J].Current Opinion in Investiga tional Drugs,2004,5(5):551-553.
[3]Iwamoto J,Takeda T,Ichimura S.Treatment with vitamin D3 and/or vitamin K2for postmenopausal osteoporosis[J].Keio Journal of Medicine,2003,52(3):147-148.
[4]王葆仁.有机合成化学(下册)[M].北京:科学出版社,1985:880.
[5]王葆仁.有机合成化学(下册)[M].北京:科学出版社,1985:565.
[6]樊能延.有机合成事典[M].北京:北京理工大学出版社,1992:757.
[7]王葆仁.有机合成化学(下册)[M].北京:科学出版社,1985:848.
[8]方朝晖,鲍陶陶,费爱华.骨疏灵对骨质疏松症大鼠骨组织形态的影响[J].中成药,2007,29(4):587-588.
[9]谢 文,冯 坤,陈宝龙.中药对实验性骨质疏松大鼠骨骼影响的形态计量学研究[J].中医正骨,2007,19(3):7-8.
[10]王立英,蔡跃增.骨形态结构的影像学研究方法[J].国外医学临床放射学分册,2007,30(2):115-118.
[11]章明放,张乃鑫,谭郁彬,等.周期性与连续性给予羟乙磷酸钠对去势大鼠骨质疏松症治疗作用的比较[J].中国骨质疏松杂志,1995,1(2):104-107.
[12]J.J.Cees.Tack,Paul Smitts.Troglitazone Decreases the Pro p o rtion of Small,Dense LDL and Increases the Resistance of LDL to Oxidation in Obese S u b j ect [J].E m e r g i n g T r e a t m e n t s a n d T e c h n o l o g i e s,1998,21(5):796-799.
[13]庞建华,张耀苏,杰英.罗格列酮和二甲双肌对高糖高脂诱导的糖尿病脂肪肝大鼠的影响[J].肝脏,2007,112(5):378-381.
[14]常保成,郑少雄.胰岛素增敏剂——罗格列酮在糖尿病中的应用[J].国外医学内分泌分册,2001,2(1):34-36.
[15]王霜,杨正国.降糖新药罗格列酮研究进展[J].临床荟萃,2002,17(7):432-433.
[16]朱波,薛耀明,李晨钟等.罗格列酮对OLETF大鼠胰岛β细胞凋亡水平的影响[J].中国糖尿病杂志,2007,15(1):44-46.
[17]文金生,胡弼,龙光.罗格列酮对果糖饲养大鼠血糖血脂及血管重构的影响[J].中国药理学通报,2004,20(5):571-575.
[18]杨晓峰,李升刚,于得志,等．复方地骨皮冲剂对小鼠血糖血脂及免疫功能的影响[J]．齐鲁医学杂志,2000,15(2)：84-86．
[19]D.M.Ribnickya,A.Pouleva,M.Watfordb.Antihyperglycemic activity of TarralinTM,an ethanolic extract of Artemisia dracunculus L [J].Phytomedicine,2006,13:50-557.
[20]M.M.Ma,J.L.Chen,G.G.Wang,et al.Sphingosine kinase 1 participates in insulin signaling and regulates glucose metabolism and homeostasis in KK/Ay diabetic mice[J].Diabetologia,2007,50:891-900.
[21]Hayato Maeda,Masashi Hosokawa,Tokutake Sashima.Dietary Combination of Fucoxanthin and Fish Oil Attenuates the Weight Gain of White Adipose Tissue and Decreases Blood Glucose in Obese/Diabetic KK-Ay Mice[J].Agric.Food Chem,2007,55:7701-7706.
[22]N.Irwin,P.L.McClean,F.P.M.O'Harte.Early administration of the glucose-dependent insulinotropic polypeptide receptor antagonist (Pro3) GIP prevents the development of diabetes and related metabolic abnormalities associated with genetically inherited obesity in ob/ob mice[J].Diabetologia,2007,50:1532-1540.