抗肿瘤17_噻唑烷酮取代甾体衍生物的仿生合成

化学Biomimetic synthesis of anti neoplastic 17 thiazolidine substituted steroidsM ENG Ge,LI Z henyu,W A NG M ei(Faculty o f P ha rmacy,School o f M edicine,Xi'an Jiaoto ng U niv ersit y,Shaanx i Xi'an,710061,China)Abstract:Objective T o o bt ain new 17 substituted ster oidal derivativ es as the possible anti neoplastic drug candidates.M ethods 17 (1,3 thiazolidinedione 5 yl) stero idal bioconjug ate w as designed by the method o f scaffo ld hopping and was sy nt hesized fro m co m mer cially av ailable mater ial via thr ee efficient steps including one bio mimetic st ep of reactio n.Result Both the inter mediate and tar g et molecule w ere r epo rted for the f irst time.T he structures of the all these new com pounds have been ver ified by spectr al analy sis,such as I R,1H NM R,13C NM R ,M S and element al analysis.Co nclusion T he sy nt hetic method and drug like pr operties of t he targ et compound mig ht be pro mising fo r further development.Key words:ster oid;t hiazo lidinedione;knoevenagel co ndensat ion;bio mimetic reduct ion;anti neoplastics抗肿瘤17 噻唑烷酮取代甾体衍生物的仿生合成孟 歌,李震宇,王 梅(西安交通大学药学系药物化学教研室,陕西西安710061)摘要:目的 合成17 取代甾体衍生物,寻找具有抗肿瘤活性的先导化合物。

方法 通过骨架迁跃的方法设计了一种结构新型的17 噻唑烷酮取代甾体生物缀合物,目标化合物由商业易得的原料经三步有效反应合成得到,其中涉及到一步生物催化的仿生合成。

结果 所得目标化合物和中间体未见文献报道,所有新化合物的化学结构经IR,1H N M R,13C N M R,M S 和元素分析等波谱解析验证。

结论 该噻唑烷酮 甾体生物缀合物具有一定的类药性,且合成方法简单,值得进一步的结构修饰和开发。

关键词:甾体骨架;噻唑烷二酮;脑文阁缩合;仿生合成;抗肿瘤do i:10.3969/j.issn.1004 2407.2011.04.027中图分类号:R914 文献标识码:A 文章编号:1004 2407(2011)04 0288 04基金项目:西安交通大学博士启动基金(2007)Large evidence show s that steroids and their bioconjug ates have a broad spectrum of bio logical activities,such as potent antimicrobial,analgesic,an ti inflam matory and anti tumo r activities[1 3].Themino r chang es in steroid structure could cause ex tensive changes in their bio logical activ ity and co n sequently in their therapeutic applications.Due to their lipophilic characters,m em br ane affinity and the ability o f binding to low density lipo pro teins,de sign and synthesis o f steroidal bioconjug ates are be coming mor e and mo re important.It is w orthy to be mentioned that 17 modification on the steroid skele to n o ften increases the bio logical activity,w hich have been v er ified in the cases o f solacongestidine,to matidine,solasodine,o lanocapsine and v erazine [4].Bearing a five member ring on the 17 position of the ster oid skeleton (Fig.1),cardiac g lycoside or cardiac g lycogenin show s the activ ity against m any kinds of tum or [5 6].Cardiac g lyco side,like ouabain and dig itox igenin,mig ht induce the apo to sis of hu m an androgen independent pro state cancer cells [7 8].As the w ater so luble ex tr actio n from Ner ium indicum M ill and the first patented car diac g lycosideanticancer drug appro ved by FDA [9],Anvirzel co n tains tw o m ain active ingr ediants ar e oleandr in [10]and oleandrigenin [11]and has now passed the clinicalphase [12].A Chinese patent also claimed the anti tumor activity and preparation of g lycosides com pounds [13].Fig.1 Contrast between the structures of cardiac glycoside Aand the target moleculeT he ear ly research also sug gested that a new ly discovered compound w ith car diac g lycoside A type skeleton,separated from traditio nal Chinese medi cine show s m oderately antineoplastic activity [2].W hile co nsidering the antineoplastic activity of the thiazolidinedione containing heterocyclic com pounds [14]and the similarity betw een the thiazolidin edione fiv e m em ber ring and the lactone five m em ber r ing on 17 position o f cardiac gly coside (Fig.1),w ehave designed a br and new 17 thiazolidinedione methy lene substituted steroid bioconjug ate in order to increase the lipophilicity and the enhance per mea bility through cell m em br anes and favo rable specific org an distribution and so to enhance its antineo plas tic activity.T he synthesis of this compound w as achieved by three steps of efficient reactio ns,inclu ding of heterocyclic cy clization,Knoevenag el con densation and reduction starting from the readily a vailable materials such as chloroacetic acid,thiourea and dehydr oepiandrosterone (2)(Fig.2).Fig.2 Synthetic route of the target moleculeExperimentalMelting points w ere taken on a WRS 1digital melting point apparatus and w ere uncorr ected.Ele m ental analyses w ere perform ed on a Carlo Elba 1106elemental analy zer.IR spectra w ere recorded on a Nicolet FI IR 360Spectrophotometer.1H NM R spectra w ere on a Bruker AM 400(400MH z)spec tro meter w ith T MS as an inter nal standard.Chemi cal shifts w ere r eported in ppm dow nfield ( ).M ass Spectra w as measured o n a H P5988A instrument by direct inlet at 70eV.All m aterials w ere obtained fro m com mercial suppliers and used as received.Synthesis of 2,4 thiazolinedione(1):1w as synthesis fro m thiourea and choloracetic acid acco rding to li teratur e [15 16].Synthesis of (5Z) 5 ((10R,13S ) 1,3,4,7,8,9,10,11,12,13,15,16 dodecahydro 3 hydroxyl 10,13 dim ethyl 2H cyclopenta[ ]phenanthren 17(14H ) yli dene)thiazolidine 2,4 dione (3):T o a 50mL three neck flask equipped w ith a thermo meter and a co n denser,w as added under the magnetic stirring of (10R,13S ) 1,3,4,7,8,9,10,11,12,13,15,16 dodeca hy dro 3 hy dro xy 10,13 dim ethyl 2H cyclopenta [ ]phenanthren 17(14H ) o ne (2,1.16g,4mmo l)and 2,4 thiazolinedione (1,0.47g ,4mm ol),and then the catalytic am ount o f anhydr ous piperidine (0.058mL,0.06mm ol)w as added by a micro sy r ing e.All these abo ve material w as dissolved by etha no l (20mL )as so lvent co mpletely w hile being stirred.T he r eaction mix tur e w as heated to reflu x ing and w as monitored by TLC.After refluxing for 7h,tw o of the m ain m aterials disappear ed,the r eac tion m ixture w as co oled dow n to roo m temperatur e.Distilled w ater (10mL)w as added dro p w isely un der v ig oro us stirring ,a large amount of w hite flo ccu lation pr ecipitation w as form ed in the formerly o paque reaction mixture.The white product w ash ob tained by filtration under vacuum,washed with distilled w ater,dried in vacuo at room temperature,w hite solid w as then obtained as the crude compound 3(1.21g,78.1%).This compound could be further purified by column chrom atog raphy (Stationary:Silica g el,m o bile:petroleum ether and EtOA c:2 1,R f =0.4)to g et a w hite pow der (0.80g,51.4%).The cr ude compound could also be r ecrystallized fro m ethy l ac etate to give a w hite cry stal like so lid for larg e scale r eaction w ith the y ield of 60%.mp 150 152 ,IR (KBr ):3431( N H ),1740( C=O ),1728( C=O )cm;1H NMR (CDCl 3): :0.89(s,3H ,CH 3),1.04(s,3H ,CH 3),1.04 2.48(m,20H ,steroid skeleton CH ),3.53(s,w,1H ,OH ),5.39(s,1H ,C=CH 6);13C NMR (CDCl 3): :22.3,22.9,23.3,26.4,30.1,30.2,31.9,32.1,37.2,37.5,37.9,42.0,47.6,51.0,66.7,71.8,113.1,122.0,141.0,165.0,167.2,177.4.MS (m/z ,%):388(M +1),Anal.Cald for C 22H 29NO 3S (%):C,68.18;H ,7.54;N,3.61;S,8.27.Found:(%):C,67.16;H ,7.49;N ,3.58;S,8.33.Synthesis of 5 ((10S,13S ) hexadecahydro 3 hydroxy 10,13 dimethyl 1H cyclopenta [ ]phenanthren 17 yl)thia zolidine 2,4 dione (4):T o a beaker added g lucose (2g,11mmo l)and w ater (40mL)and the dried baker 's yeast pow der (100mg ),the m ix tur e w as cultivated at ro om tem perature for 30minutes.T o a 100mL three neck flask equipped w ith a g lass inlet of g as w as added 3(200m g,0.52mm ol),1,4 diox ane (10m L )and the abo ve yeast cultiv atedmixture.T he reaction mixture w as first intr oduced w ith N2and then H2.The reactio n m ixture w as stirred under hydrog en atm ospher e.T LC m onitor w ith the elution of petro leum/ethyl acetate(3 1) show ed that the reduction completed after48hours under room temperature.The reaction co uld also be com pleted after24hour s under38 .The reaction mixture w as filtered to get rid of insoluble solid. The filtr ation w as concentrated under vacuo to g ive an oily crude pro duct as the crude targ et mo lecule4. This com po und w as pur ified by co lum n chr omatog r aphy(Stationary:Silica gel,mobile:petr oleum ether and EtOAc:2 1,R f=0.5)to give pure target com po und4(186mg,48mmo l,92%).m p149 151,IR(KBr):3389( N H),2816(C H ),C=O1739(C=O ),1672(C=O)cm-1;1H NM R(CDCl3): :1.06(s,3H,CH3),1.16(s,3H,CH3),1.24 1.98 (m,23H,steroid skeleton CH),3.16(m,1H,3 CH OH),3.68(s,w,1H,OH),9.8(s,1H, NH);13C NMR(CDCl3): :15.3,16.2,20.9,23.4,26.9,29.1, 31.0,32.2,32.7,35.8,36.9,37.2,39.4,41.6,44.7, 49.4,51.8,52.8,55.8,71.5,167.1,174.6.M S(m/ z,%):392(M+1),Anal.Cald fo r C22H33NO3S (%):C,67.48;H,8.49;N,3.58;S,8.19.Fo und: (%):C,67.39;H,8.46N,3.53;S,8.18.Drug like properties prediction:Lipinski's rule of five is a simple m odel to forecast the absorptio n and in testinal perm eability of a compound[17 18].In the rule of fiv e model,co mpo unds are considered likely to be w ell absor bed w hen they possess these attri butes m olecular w eig ht<500,lg P<5,num ber of H bond donors<5,num ber H bond acceptor s<10, and number of rotatable bonds<10.T he dr ug like parameter s o f both3and4w e calculated via Chem draw softw are(Cambridge3D8.0)(T able1).All com po unds meet the drug like criteria of rule of five.Co nnolly m olecular ar ea(M S)w as obtained via Chem3D U tra(Cam br idge softw are)according to the follow ing procedure:mo lecular dy namics calcu lation and energy minimization w ere sequentially run on each mo lecule w ith default values(step interv al = 2.0fs,frame interval=10fs,terminate after10 000steps),the parameter s of com po unds(3and4) w ere then co mputed,respectively.Table1 Biological Prediction of the properties of the target moleculesCompounds34Properties Criteria M W387.546391.578<500lg P 3.0414 4.1286<5 Connolly molecular area( 2)334.318332.346Number of H bond donors22<5 Number of H bond acceptors32<10 Number of rotatab le bonds34<10ConclusionsAs a sum mary,tw o new stero id bioconjug ates as the antineoplastic ag ents hav e been desig ned and synthesized via biom imic w ay from comm er cial a vailable materials.The new ly reported3and4w er e first obtained by Knoevenag el condensatio n and the biomim etic reduction,the former metho d is a mor e efficient to intr oduce C=C double bo nd to the17 posito n of the ster oid skeleton for its sim ple w ork ups and relative hig h y ield com paring w ith other tra ditional methods such as W ittig reaction,etc.All o f the new com pounds w ere pr edicted from their drug like param eters,such M S,lg P,etc.All o f the com pounds(3,4)hav e lg P value o f<5.T herefore,bo th 3and4have show s the favo rable drug like pr oper ties and are potentially interesting for further opti mization.AcknowledgmentsT he research w ork w as funded by Xi'an Jiao tong University Docto r Initiating Fund(2007). 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