Benzylated Dihydroflavones Thiab Isoquinoline-Drived Alkaloids Los Ntawm Cov tawv ntoo ntawm Diclinanona Calycina (Annonaceae) Thiab Lawv Cytotoxicities

Yog xav paub ntxiv, hu rautina.xiang@wecistanche.com

Abstract: Diclinanona calycina RE Fries nrov npe hu ua "enviro", yog ib hom ntawm Annonaceae tsev neeg nyob rau Brazil. Hauv peb qhov kev tshawb nrhiav tsis tu ncua rau bioactive compounds los ntawm Annonaceae Amazon cov nroj tsuag, cov tawv ntoo ntawm D.calycina tau tshawb xyuas los ntawm cov txheej txheem chromatography classical uas tau muab kaum peb lub tebchaw (alkaloids thiab flavonoids) tau piav qhia thawj zaug hauv D. calycina thiab hauv cov genus Diclinanona. Cov qauv ntawm cov khoom sib cais no tau tsim los ntawm kev tshuaj xyuas dav siv 1D / 2D-NMR spectroscopy ua ke nrog MS. Cov alkaloids cais tau raug txheeb xyuas raws li cov subclasses: yooj yim isoquinoline, thali online (1); aporphine, anonaine (2); oxoaporphine, liriodenine (3); benzyltetrahydroisoquinolines, (S)-(ntxiv)-reticuline(4); dehydro-oxonorreticuline (3,4-dihydro-7-hydroxy-6-methoxy-1-}isoquinolinyl)(3-hydroxy-4-methoxyphenyl)-methanone)(5 );(plus )-1S,2R-reticuline Ng-oxide (6); thiab (ntxiv rau) -1S,2S-reticuline N -oxide (7); tetrahydro protoberberine, coreximine(8); thiab pavine, bisnorargemonine (9). Thaum lubflavonoidsbelongs rau benzylateddihydroflavones, isorhamnetin (10), dichamanetin (11), thiab kev sib xyaw ntawm uvarinol (12) thiab isouvarinol (13). Compound 5 tau piav qhia thawj zaug hauv cov ntaub ntawv raws li cov khoom ntuj tsim. Kev ua haujlwm cytotoxic ntawm cov ntsiab lus sib cais tau raug tshuaj xyuas tiv thaiv kab mob qog noj ntshav thiab cov kab mob uas tsis yog mob qog noj ntshav. Ntawm cov tshuaj ntsuam xyuas, cov txiaj ntsig tau pom zoo tshaj plaws tau pom rau benzylated dihydroflavones dichamanetin (10), thiab sib xyaw ntawm uvarinol (12) thiab isouvarinol (13), uas tau nthuav tawm cov kev ua haujlwm cytotoxic nruab nrab tiv thaiv kev sim.mob qog noj ntshavkab(<20.0 ug·ml-)="" and="" low="" cytotoxicity="" against="" the="" non-cancerous="" cell="" line="" mrc-5(="">25.0 ug·mL-I). Dichamanetin (11) tau qhia cytotoxic kev ua haujlwm tawm tsam HL-60 thiab HCT116 nrog IC50 qhov tseem ceeb ntawm 15.78 ug·mL-I (33.70 umol·LI) thiab 18.99 ug∶mL-(40.56 umol. L-), raws li qhov sib xyaw ntawm uvarinol (12) thiab isouvarinol (13) tau pom cytotoxic kev ua haujlwm tawm tsam HL-60, nrog rau IC5 tus nqi ntawm 9.74 ugs.mL-1, thiab HCT116, nrog IC50. 17.31 Nws. mL- ib. Cov kev ua cytotoxic no tuaj yeem raug ntaus nqi vim muaj ib lossis ntau dua hydroxy benzyl pawg tam sim no nyob rau hauv cov molecules nrog rau txoj hauj lwm ntawm cov pab pawg no txuas. Cov kev ua cytotoxic ntawm reticuline, anonaine, thiab liriodenine tau tsim yav dhau los, nrog liriodenine yog cov tshuaj muaj zog tshaj plaws.

Ntsiab lus: Diclinanona calycina; alkaloids thiab benzylated dihydroflavones; cytotoxic kev ua haujlwm

Nyem kom paub ntau ntxiv txog cov khoom

1. Taw qhia

Annonaceae yog ib tsev neeg loj ntawm cov ntoo kub thiab subtropical ntoo thiab tsob ntoo, muaj txog 112 genera thiab 2440 hom [1]. Ntau hom paub txog lawv cov txiv hmab txiv ntoo thiab cov khoom siv tshuaj [2]. Kev tshawb nrhiav phytochemical yav dhau los nrog qee hom Annonaceae coj mus rau kev sib cais thiab cov yam ntxwv ntawm cov chav kawm sib txawv ntawm cov metabolites, xws li monoterpenes, diterpenes, triterpenes, lignans,flavonoids, asarone-derived phenylpropanoids, acetogenins, thiab feem ntau yog isoquinoline-derived alkaloids [3-7]. Qee yam ntawm cov metabolites thib ob cais tawm los ntawm hom Annonaceae tau nthuav tawm cov haujlwm tseem ceeb hauv kev lom neeg, xws li tshuaj tiv thaiv kab mob thiab urease-inhibiting zog [8,9], trypanocidal [10,11], leishmanicidal [11,12], antimalarial [4,13] , antimicrobial [4,14,15], antioxidant thiab antirheumatic ua [9,15] thiab, tshwj xeeb tshaj yog, kev ua cytotoxic tawm tsam tib neeg qog cell kab [4,6,11,16-21].

Txawm hais tias tsev neeg Annonaceae suav hais tias yog tsev neeg keeb kwm thiab kawm tau zoo, qee qhov kev tshawb fawb phytochemical thiab / lossis pharmacological tau ua nrog nws hom[3]. Phytochemical thiab / lossis kev tshawb fawb tshuaj tau tsom mus rau cov hom ntawm cov genera Annona, Asimina, thiab Cananga, vim lawv qhov tseem ceeb ntawm kev lag luam, thiab ntawm qee hom tsiaj ntawm Duguetia, Guatteria, thiab Xylopia [5]. Txawm hais tias muaj kev loj hlob zoo hauv 20 xyoo dhau los hauv kev cuam tshuam nrog kev tshawb fawb phytochemical thiab pharmacological, tus naj npawb ntawm cov tsiaj tshawb fawb tseem tsawg heev nyob rau hauv kev sib raug zoo rau ntau hom tsiaj. Tam sim no, raws li Lub Vev Xaib ntawm Kev Tshawb Fawb, Scopus, thiab SciFinder cov ntaub ntawv tshawb fawb, tsuas yog kwv yees li 15 feem pua ​​​​ntawm cov hom Annonaceae tau piav qhia muaj cov kev tshawb fawb txog phytochemical thiab / lossis pharmacological.

Ntawm cov me-studied hom yog cov genus Diclinanona Diels. Cov genus no belongs rau pawg neeg Annoneae, ntawm subfamily Annonoideae, thiab tshwm sim tsuas yog nyob rau hauv teb chaws sov South America (tsuas yog hauv thaj av Amazon). Nws yog ib tug genus uas muaj tsuas yog peb hom, Diclinanona calycina(Diels)RE Fries Diclinanona matogrossensis Maas thiab Diclinanona tessmannii Diels, uas tshwm sim li ntoo [22-24].D.calycina (synonymy Xulopia calycina Diels) yog ib tug 8 rau 30 m siab ntoo, nrov npe hu ua "envireira" thiab "enviro", faib thoob plaws Amazon phiab hauv Brazil, Peru, thiab Venezuela [23]. D.calycina yog superficially zoo ib yam li Xylopia rau nws cov paj nrog elongate thiab nqaim petals, tab sis nws txawv rau nws woody, indehiscent, globose, thiab tuab-walled monocarps [23,24].

Cov kev tshawb fawb yav dhau los nrog D. calycina qhia tsuas yog kev tshawb fawb tshuaj. Thawj txoj kev tshawb fawb piav qhia txog kev tshawb nrhiav cov tshuaj tua kab mob ntawm cov tshuaj methanolic, chloroform, thiab cov tshuaj aqueous tawm tsam cov kab mob Mycobacterium smegmatis, Escherichia coli, Streptococcus sanguis, Streptococcus oralis, Staphylococcus aureus, thiab Candida albicans siv txoj kev 2. Qhov thib ob qhia txog kev tshawb xyuas ntawm cov tshuaj tiv thaiv kab mob ntawm cov organic (dichloromethane: methanol 1: 1) thiab cov tshuaj aqueous tawm tsam cov kab mob Enterococcus faecalis siv microdilution broth assay (MDBA) thiab disk diffusion assay (DDA) [26]. Yog li, hauv peb qhov kev tshawb nrhiav txuas ntxiv mus rau cov khoom siv ntuj tsim tshiab los ntawm Annonaceae los ntawm Amazon rainforest, txoj kev tshawb no tsom mus soj ntsuam cov phytochemical thiab pharmacological zog ntawm cov tawv ntoo ntawm D.calycina. Hauv daim ntawv tshaj tawm no, kaum peb lub tebchaw (cua alkaloids thiab plaub benzylateddihydroflavones) tau raug cais tawm thiab txheeb xyuas thawj zaug hauv D.calycina, nrog rau hauv cov genus Diclinanona. Ntxiv rau, cytotoxicity ntawm cov ntsiab lus tseem ceeb tau tshawb xyuas tawm tsam B16-F10, HepG2, K562, thiab HL{{ 4}} qog cell kab siv Alamar xiav tshuaj ntsuam.

2. Cov txiaj ntsig thiab kev sib tham

2.1.Structural Elucidation of the Compounds

Tom qab tshawb pom cov muaj nitrogen-muaj cov tebchaw hauv cov tshuaj methanolic uas siv Dragendorff's reagent, nws tau raug kev kho cov kua qaub raws li cov txheej txheem ntawm Costa li al. [12] ua rau alkaloidal thiab nruab nrab feem. Ib tug siab concentration ntawm nitrogen-muaj cov tebchaw tau pom nyob rau hauv lub alkaloidal feem uas raug rau chromatographic tsom xam. Kev sib cais ntawm chromatographic ua tiav, raws li tau piav qhia hauv ntu Extraction thiab cais tawm, tau coj mus rau kev sib cais thiab kev txheeb xyuas ntawm kaum peb cov tshuaj lom neeg (1-13, Daim duab 1), cuaj isoquinoline-derived alkaloids 1-9, thiab plaub benzylated dihydroflavones 10-13. Cov qauv ntawm cov khoom sib cais no (Daim duab 1) tau tsim los ntawm kev tshuaj xyuas dav siv 1D thiab 2D NMR spectroscopy ua ke nrog MS (Cov Ntaub Ntawv Ntxiv), nrog rau kev sib piv nrog cov ntaub ntawv los ntawm cov ntaub ntawv.

Compound 5 tau txais los ua xim av amorphous hmoov thiab kuaj pom zoo rau Dragendorff's reagent. Nws pom ib qho protonated molecule ntawm m/z328 [M ntxiv rau H] 'hauv LR-ESI (ntxiv rau) MS-tshaj nrog cov mis molecular C18H1NO5. Lub 'H thiab 13C-NMR spectra ntawm 5 (Table 1) tau zoo ib yam nrog cov reticuline (4) [27, tshwj tsis yog rau qhov tsis muaj nitrogen-bonded methyl pawg (CH3-N) thiab cov teeb liab ntawm pawg methine nyob rau hauv txoj hauj lwm 1, uas tau hloov los ntawm lub teeb liab ntawm δc 165.1 nyob rau hauv ib txwm 13C-NMR spectrum ntawm pawg imine conjugated rau ib pawg carbonyl ntawm bc 192.8. Hauv 'H-NMR spectrum, peb downfield hydrogens atδH 6.88 (1H,d, J=8.4 Hz, H-5'), 5H7.57(1H,d, J{{33}) } . , nthuav tawm lub xub ntiag ntawm ABX coupling system. Ob lub cim qhia ntawm 6H 6.90 (1H, s, H-8) thiab 6.71(1H, s, H-5) qhia tias muaj 1,2,4,5-tetrasubstituted phenyl nplhaib. Tsis tas li ntawd, H-NMR spectrum ntawm 5 kuj pom cov cim rau ob pawg methoxy resonating ntawm bH 3.93 (3H, s) thiab 6H 3.95 (3H, s), thiab cov teeb liab rau ob pawg methylene resonating ntawm δμ 2.79 (2H, t, J=7.8 Hz) thiab 6H 3.89(2H, t,/=7.8 Hz), ntaus nqi rau H-4 thiab H-3, raws li (Table 1) . Cov ntaub ntawv no tau qhia tias alkaloid 5 muaj benzyltetrahydroisoquinoline skeleton [28-30].

Cov pab pawg no tau tsim los ntawm ob thiab peb daim ntawv cog lus 'H-13Ccorrelation map los ntawm HMBC-NMR kev sim (Daim duab 2 thiab Table 1). Qhov kev tshuaj ntsuam no tau qhia tias cov hydrogen ntawm δH 3.89 (H-3) qhia peb-band 'H-13C sib raug zoo nrog cov carbons ntawm 6c130.1(4a) thiab δc165. {42}} (C-1) thiab ob daim ntawv cog lus lH-13C sib raug zoo nrog cov pa roj carbon atδc 25.4(C-4), lees paub tias muaj pawg imine hauv cov molecule. Ntawm qhov tod tes, cov cim ntawm δH 7.57 (H-2') thiab δH 7.60 (H{27}}') qhia peb-bond lH-13C sib raug zoo nrog cov carbons ntawm δc 124.3( C-6'),5c 151,4(C-4') thiab δc192.8(C-7), and5c116.0(C-2'),6c 151.4 (C-4') thiab c 192.8(C-7'), feem, tsim cov carbonyl pawg hauv cov molecule (Daim duab 2 thiab Table 1). Yog li ntawd, raws li cov ntaub ntawv NMR no, compound 5

tau tsim los ua benzyltetrahydroisoquinoline alkaloid 34-dihydro-7-hydroxy-6-methoxy-1- isoquinolinyl)(3-hydroxy-4-methoxyphenyl)-methanone, uas yog npe hu ua dehydro-oxonorreticuline. Cov alkaloid no tau piav qhia thawj zaug hauv cov ntawv nyeem raws li cov khoom siv ntuj tsim. Nws thawj zaug thiab tsuas yog cov ntaub ntawv tau piav qhia los ntawm Dörnyei li al. xyoo 1982 【31】 raws li cov khoom siv hluavtaws. Tsuas yog cov ntaub ntawv 'H-NMR tau piav qhia nrog qee qhov tsis tau txiav txim siab ntau yam. Yog li, cov haujlwm ua tiav rau tag nrho 1H- thiab 13C-NMR tshuaj hloov pauv tau tsim los ntawm ib daim ntawv cog lus (HSOC) thiab ob thiab peb daim ntawv cog lus (HMBC) 'H-13C-NMR kev sib raug zoo sim, thiab tau piav qhia hauv Table 1.

Cov tshuaj 6 thiab 7 tau txheeb xyuas tias yog benzyltetrahydroisoquinoline alkaloids (ntxiv rau) -1S,2R-reticuline-N -oxide, thiab (ntxiv rau)-1S,2S-reticuline-N -oxide, raws li. Cov ntaub ntawv H-NMR ntawm cov alkaloids no tau muab piv nrog cov ntaub ntawv piav qhia los ntawm Lee li al. [28] siv tib yam deuterated hnyav (CD3OD) thiab qee qhov tsis sib xws tau pom (Table 1), feem ntau yog cuam tshuam nrog H-1, H-8, thiab H, C-NO. Tsis muaj 13C-NMR cov ntaub ntawv piav qhia rau cov alkaloids no hauv cov ntaub ntawv. Raws li qhov txwv 1H- thiab 13C-NMR cov ntaub ntawv, nrog rau qhov tsis meej meej pom rau cov molecules, 'H thiab 13C 1D thiab 2D NMR kev sim tau ua los txiav txim siab lawv txoj haujlwm raug thiab ntau yam.

Qhov tseeb txoj hauj lwm ntawm hydrogen H{{0}} thiab H-1 ntawm ob isomers 6 thiab 7 yog tsim los ntawm peb-bond 'H-13C correlation daim ntawv qhia los ntawm HMBC -NMR kev sim (Daim duab 2). Rau cov isomer 6 qhov kev tshuaj ntsuam tau qhia tias hydrogen ntawm δH 5.78 (H-8) qhia peb-bond 'H-13C sib raug zoo nrog cov carbons ntawm 6c 79.8 (C-1), δc120 .6(4a), thiab δc 149.4 (C-6), thaum lub hydrogen ntawm δH 4,13 (H-1) qhia peb-bond 'H-13C correlation nrog cov carbons ntawm c 60.7 (C-3), 6c 115.7 (C-8), δc 120.6 (C-4a), thiab 131.2 (C-1'), yog li tsim cov txoj haujlwm raug ntawm cov hydrogen H-8 thiab H-1 rau isomer 6. Qhov no tuaj yeem raug lees paub ntxiv los ntawm kev txheeb xyuas cov hydrogen ntawm δH 6.76 (H-5) uas pom peb daim ntawv cog lus H-13C sib raug zoo nrog cov carbons atδc 27.0 (C-4), δc 127.1 (C-8a), thiab δc 145.7 (C-7) (Figure2). Rau cov isomer7 qhov kev tshuaj ntsuam tau qhia tias hydrogen ntawm δH 6.30 (H-8) qhia peb-bond 'H-13Ccorrelation nrog cov carbons ntawm δc 79.4 (C-1), δc 122.8 ( 4a), thiab δc 148.9 (C-6), thaum cov hydrogen ntawm δH 4.46 (H-1) qhia peb-bond H-13C sib raug zoo nrog cov carbons ntawm δc 62.9 (C{ {82}}), 5c 115.5 (C-8), bc 122.8 (C-4a), thiab 131.4 (C-1'), yog li tsim kom muaj txoj haujlwm raug ntawm hydrogen H -8 thiab H-1 rau isomer 7. Qhov txiaj ntsig no kuj tau lees paub los ntawm kev txheeb xyuas cov hydrogen ntawm δH 6.71 (H-5) uas qhia peb-bond lH-13C Kev sib raug zoo nrog cov carbons ntawm δc26.3 (C-4), δc 126.9 (C{106}}a), thiab 6c 146.1 (C-7) (Daim duab 2). Qhov kev sim NOESY kuj tau ua los tsim kom muaj qhov tseeb stereochemistry ofisomers6 thiab 7. Hauv qhov kev sim no, NOE muaj zog sib raug zoo ntawm H-1(δH 4.13) thiab H, CN({118}}.15) qhia txog qhov -orientation ntawm cov pa oxygen hauv isomer 6. Ntawm qhov tod tes, tsis muaj qhov pom tseeb NOE kev sib raug zoo ntawm H-1(6H 4.46) thiab HAC-N(5H 3.20) tuaj yeem pom hauv 2D-NOESY kev sim rau 7, qhia txog -orientation ntawm cov pa hauv isomer 7 (Daim duab 2). Cov kev sib txawv me me ntawm cov tshuaj hloov pauv ntawm cov isomers no tau pom meej meej vim yog qhov stereochemistry ntawm nitrogen cuam tshuam los ntawm cov haujlwm thiab cov haujlwm ntawm oxygen. Kev sib piv ntawm 'H-thiab 13C-NMR cov ntaub ntawv tau txais rau cov tebchaw 6 thiab 7 nrog cov ntaub ntawv ntawm cov molecules nrog cov qauv ze xws li hexapetaline A thiab hexapetaline B [30] txhawb nqa cov ntaub ntawv tau piav qhia hauv Table 1 yam tsis muaj qhov tsis meej.

Cov tshuaj sib xyaw ua ke 1-4 thiab 8-13 tau txheeb xyuas tias yog thalifoline (1) [32], anonaine (2) [33] liriodenine (3)[10,34], (S)-( ntxiv rau )-reticuline ( 4) [27], coreximine (8) [34,35], bisnorargemonine (9) [36], isochamanetin (10) [37], dichamanetin (11) [37] thiab kev sib xyaw ntawm uvarinol (12) thiab isouvarinol (13) [37] raws li lawv cov spectroscopic profiles thiab kev sib piv nrog cov nqi hauv cov ntaub ntawv. Lub lH-thiab 13C 1D thiab 2D-NMR spectra, nrog rau qhov loj ntawm txhua qhov sib cais, muaj nyob hauv Cov Khoom Siv Ntxiv.

Los ntawm chemophenetic (ib lo lus tshiab rau cov nroj tsuag chemosystematics / cog chemotaxonomy) taw tes ntawm view, nws yog ib qho tseem ceeb kom nco ntsoov tias muaj C-benzylated flavanones thiab C-benzylated dihydrochalcones yog ib hom tshwj xeeb ntawm flavonoids muab los ntawm flavanone pinocembrin. thiab tau piav qhia tshwj xeeb hauv hom ntawm genus Uoaria teej tug mus rau tsev neeg Annonaceae [3,38-42]. Lub xub ntiag ntawm cov tebchaw no hauv D. calycina qhia txog kev sib raug zoo chemophenetic nrog Uoaria. Ntawm qhov tod tes, kev tshawb nrhiav ntxiv yuav tsum tau ua nrog rau lwm qhov ntawm cov nroj tsuag, nrog rau lwm hom Diclinanona kom paub meej tias qhov kev sib raug zoo chemophenetic no. Flavanones thiab chalcones tau dav dav hauv cov nroj tsuag siab dua, tab sis qhov sib ntxiv ntawm benzyl pawg yog qhov tsawg heev thiab zoo li tsis tshua muaj nyob hauv Annonaceae rau genus Uoaria [3,39-42] thiab tam sim no mus rau genus Diclinanona. Benzyl pawg ntseeg tau tshwm sim los ntawm C6-C1 txoj hauv kev, tab sis o-hydroxy functionality yog txawv. Qhov tsis muaj cov hloov pauv hauv B-ntiv nplhaib hauv tag nrho cov flavonoids ntawm Uoaria thiab Diclinanona tuaj yeem txuas nrog kev soj ntsuam yav dhau los txog cov flavonoids ntawm Popowia cauliflora [38].

Cov isoquinoline-derived alkaloids cais thiab piav qhia hauv txoj haujlwm no twb tau sau npe rau ntau hom Annonaceae hauv ntau hom. Qee yam ntawm cov no, xws li liriodenine thiab anonaine, raug suav hais tias yog cov cim chemophenetic, thiab qhov muaj cov alkaloids hauv D.calycina ntxiv txhawb kev sib raug zoo ntawm cov cim chemophenetic hauv tsev neeg Annonaceae [5-7,15,20,27, {{5},43].

Nws yog tsim nyog hais tias, raws li Zidorn [44], chemophenetic cov kev tshawb fawb tau txhais raws li kev tshawb fawb tsom los piav qhia txog cov array ntawm cov metabolites tshwj xeeb theem nrab hauv ib daim ntawv teev npe, raws li twb tau pom nyob rau hauv ntau cov ntawv luam tawm [5-7,15,20 ,27,{5}},43-46]. Yog li, cov kev tshawb fawb chemophenetic pab txhawb rau cov lus piav qhia phenetic ntawm taxa, zoo ib yam li anatomical, morphological, thiab karyological mus kom ze, uas twb tau lees paub tias yog qhov tseem ceeb heev rau kev tsim cov "natural" systems, thiab tseem yog qhov tseem ceeb heev rau kev lag luam. Kev piav qhia ntawm cov kab mob sib cais nrog kev pab ntawm cov txheej txheem molecular niaj hnub [44].

2.2. Cytotoxic Assay

Kev ua haujlwm hauv vitro cytotoxic ntawm cov tshuaj sib cais (Table 2), tshwj tsis yog cov tebchaw 2, 3,5, thiab 8 (vim lawv cov txiaj ntsig qis), raug tshuaj xyuas los ntawmmob qog noj ntshavkab HL-60 (tib neeg promyelocytic leukemia), MCF-7(tib neeg lub mis adenocarcinoma), HepG2 (human hepatocellular carcinoma), HCT116 (tib neeg txoj hnyuv kab mob), thiab kab mob uas tsis yog mob qog noj ntshav MRC{{5} }( tib neeg lub ntsws fibroblast) siv Alamar blue assay tom qab 72 teev ntawm incubation.

Among the compounds evaluated (Table2), the most promising results were verified for benzylated dihydroflavones dichamanetin (10), and the mixture of uvarinol (12)and isouvarinol (13), which showed moderate cytotoxic activity against the tested cancer cell lines and low cytotoxicity against the non-cancerous cell line MRC-5(>25.0 ug.mL-1). Dichamanetin (11) pom cytotoxic kev ua haujlwm tawm tsam HL-60 thiab HCT116 nrog IC50 qhov tseem ceeb ntawm 15.78 ug·mL-1(33.70 μmol·L-1) thiab 18.99 ug·mL-1 (40.56 umol.L-1), feem. Qhov sib xyaw ntawm uvarinol (12) thiab isouvarinol (13) tau pom cytotoxic kev ua haujlwm tawm tsam HL-60, nrog ICs0 tus nqi ntawm 9.74ug∶mL-1, thiab HCT116, nrog ICs0 tus nqi ntawm 17.31 ug∶mL{ {30}}. Ntawm cov benzylated dihydroflavones, tsuas yog isorhamnetin (10) tau pom cov tshuaj cytotoxic tawm tsam HepG2 nrog IC50 tus nqi ntawm 19.79 ugs:mL-1(54.65 umol.L-1). Raws li cov ntaub ntawv, benzylated dihydroflavones tau piav qhia nrog cytotoxic zog [39-42]. Ntawm cov uas tau piav qhia hauv txoj haujlwm no, isorhamnetin (10), dichamanetin (11), thiab uvarinol (12) tau piav qhia hauv vitro cytotoxic zog tiv thaiv tib neeg qog cell kab ntawm carcinoma ntawm nasopharynx (KB) thiab P-388 lymphocytic leukemia. (PS) nrog IC50 qhov tseem ceeb ntawm 5.3 thiab 4.1,4.8 thiab 1.8, thiab 5.9 thiab 9.7, ug.mL-1, raws li [39,40]. Cov dej num sib txawv no tuaj yeem raug ntaus nqi vim muaj ib lossis ntau dua hydroxy benzyl pawg tam sim no nyob rau hauv cov molecules nrog rau txoj hauj lwm nyob rau hauv cov pab pawg neeg no txuas. Txawm li cas los xij, kev tshawb nrhiav ntxiv yog xav tau kom paub tseeb tias qhov kev soj ntsuam no.

Cov kev ua hauv vitro cytotoxic ntawm isoquinoline-derived alkaloids anonaine (2), liri-adenine (3), thiab (S)-( plus )-reticuline (4) tsis ntev los no tau piav qhia los ntawm Menezes li al.[2{{16] }}], Souza et al. [6] thiab Costa et al. [43] nrog rau qhov tseem ceeb ntawm liriodenine, uas pom tau tias muaj zog cytotoxic kev ua haujlwm tiv thaiv kab mob qog noj ntshav B{{10}}F10(mouse melanoma), HepG2(tib neeg hepatocellular carcinoma), HL{{13} }} (tib neeg promyelocytic leukemia), thiab K562 (tib neeg mob myelocytic leukemia) nrog IC5s0 qhov tseem ceeb hauv qab no mus rau 10.0 μmol.L-1【43】. , thiab K562 nrog IC50 qhov tseem ceeb ntawm qis dua 19.0 umol.L-1[20]. (S)-( ntxiv rau )-Reticuline pom kev ua haujlwm nruab nrab tsuas yog tawm tsam HepG2 nrog IC50 qhov tseem ceeb ntawm 15.35 μmol L-1【 6, 20].

3. Cov ntaub ntawv thiab cov txheej txheem

3.1. Cov txheej txheem kev sim dav dav

Kev sib hloov kho qhov muag hauv methanol (MeOH) tau kaw nrog P{{0}} polarimeter (Jasco, Tokyo, Nyiv) ntawm 589 nm. Kev sim 1D thiab 2D NMR tau txais hauv CDCl. (chloroform-d) lossis CDCl ntxiv rau poob ntawm CD, OD (methanol-du, thiab CD, COCD3 (acetone-d6) ntawm 298 K ntawm AVANCE I HD NMR spectrometer (Bruker, Billerica, MA, USA) ua haujlwm ntawm 11.75 T ( Tes 13C ntawm 500 thiab 125 MHz, feem) thiab ntawm Bruker AVANCE I 600 NMR spectrometer ua haujlwm ntawm 14.1 T('H thiab 13Cat 60{ {63}} thiab 150 MHz feem.Txhua lH- thiab 13C-NMR tshuaj hloov pauv (δ) tau nthuav tawm hauv ppm txheeb ze rau tetramethylsilane teeb liab ntawm 0.00 ppm raws li kev siv sab hauv, thiab cov coupling constants (D) yog muab nyob rau hauv Hz. NMR spectrometer tau nruab nrog ib tug 5-mm multinuclear inverse detection sojntsuam (1D thiab 2D NMR thwmsim) nrog ib tug z-gradient. Ib-bond (HSQC) thiab ob thiab peb-daim ntawv cog lus (HMBC)1H-13C-NMR correlation thwmsim tau optimized rau nruab nrab coupling tas li' JcH thiab LRJ (cH ntawm 140 thiab 8 Hz, feem. Rau low-resolution mass spectrometry (LC-MS) soj cov qauv ntawm cov tshuaj sib cais tau rov muab tso rau hauv methanol (HPLC qib), tsim cov khoom lag luam k daws (1 mg·mL-1). Aliquots (5 uL) ntawm cov kev daws teeb meem tau ntxiv diluted mus rau 5 ug·mL-thiab tshuaj xyuas los ntawm kev tso dej ncaj qha rau hauv triple quadrupole mass spectrometer, qauv TSO Quantum Access (Thermo Scientific, San Jose, CA, USA), nruab nrog electrospray ionization (ESI) lossis atmospheric siab tshuaj ionization (APCI) qhov chaw. ESI-MS tej yam kev mob: spray voltage, 5kV; sheath gas, 10 arbitrary unit (arb); auxiliary gas, 5arb; cheb gas, 0arb; capillary temp, 250 degree; capillary voltage, 40 V; tube lens, 70 V; huab hwm coj ntau, m / z 100 txog 1000.APCI-MS tej yam kev mob: tawm tam sim no, 5 μA; vaporizer kub, 350 degree; sheath gas siab, 25 arbitrary unit (arb); ion cheb gas siab, 0.0 arb; aux gas siab, 10 arb; capillary kub, 250 ° C; tube lens offset, 70 V; skimmer offset, 0 V; huab hwm coj ntau, m / z 100 txog 1000. Argon tau siv los ua cov pa sib tsoo, thiab MS / MS spectra tau txais los ntawm kev sib tsoo energies xws li 25 txog 30 eV. Silica gel60 (Sigma-Aldrich, San Luis, MO, USA, 70-230) mesh) tau siv rau kab chromatography (CC), thaum silica gel 60 F254 (Macherey-Nagel, Düren, Lub teb chaws Yelemees, 0.25 hli, txhuas) tau siv rau kev tshuaj xyuas thiab kev npaj nrog nyias laver chromatography (PTLC) (Macherey-Nagel, 1.00 mm, iav). Cov tshuaj tau pom los ntawm kev raug UV254/365 lub teeb, los ntawm kev txau nrog p-anisaldehyde reagent tom qab cua sov ntawm lub phaj kub, thiab los ntawm kev txau Dragendorff's reagent.

3.2.Cov khoom siv cog

Hauv kev tshawb nrhiav tam sim no, cov khoom siv botanical (cov tawv ntoo) ntawm D.calycina tau sau rau 27 Tsib Hlis 2017 ntawm Adolpho Ducke Reserve (geographic coordinates: 02 degree 53'36.1S thiab 59 degree 58'28.9"W), Manaus, Amazonas State , Brazil, thiab txheeb xyuas los ntawm Prof. Dr.Antonio Carlos Webber, tus kws kos duab cog qoob loo ntawm Lub Tsev Haujlwm Saib Xyuas Kev Noj Qab Haus Huv ntawm Universidade Federal do Amazonas (DB/UFAM). nkag mus (specimen) tau sau npe hauv Sistema Nacional de Gestao do Patrimonio Genetico e do Conhecimento Tradicional Associado (SISGEN) nrog cov ntaub ntawv A70EDCD.

3.3. Extraction thiab rho tawm

Cov tawv ntoo ntawm D.calycina tau pib qhuav ntawm chav tsev kub rau 24 teev thiab tom qab ntawd qhuav hauv qhov cub cua rau 48 teev ntawm qhov kub ntawm 40 degree, thiab tom qab ntawd pulverized hauv plaub rab riam zeb grinder (Marconi, Piracicaba, SP. , Brazil) kom tau cov khoom siv hmoov (1340 g). Tom qab ntawd, ua kom tiav maceration nrog hexane (8 × 4 L) ua raws li MeOH (8 × 4 L) tau ua. Cov kev daws teeb meem uas tau txais tau yog concentrated nyob rau hauv lub evaporator teb ntawm qhov txo siab ({10}} degree) muab cov hexane (24.85 g) thiab MeOH (199.43 g) rho tawm, raws li.

TLC tsom xam nrog Dragendorff's reagent qhia tias muaj cov alka-loids hauv MeOHextract. Yog li ntawd, ib qho aliquot ntawm MeOH extract (188.30 g) yog thawj zaug raug rau cov kua qaub-base extraction kom muab alkaloidal (5.46 g) thiab nruab nrab (4.47 g) feem. Tom qab ntawd, ib feem ntawm alkaloidal feem (5.0g) tau raug silica gel chromatographic kem (CC) yav tas los kho nrog 10 feem pua ​​NaHCO3 tov [12], eluted nrog hexane (100 feem pua), hexane-CHCl, (90: 10, 80: 20, 70: 30, 60: 40, 50: 50, 40: 60, 30: 70, 20: 80, thiab 10: 90, v / o), CH2Cl2 (100 feem pua), CH2Cl 2-EtOAc(90:10,80:20,70:30,60:40,50:50,40:60,30:70,20:80, thiab 10:90,0/u), EtOAc (100%), EtOAc-MeOH(95:05, 90:10,85:05, 80:10,75:25, 70:30,60:40, 50:50,40:60, 30:70,20 :80, thiab 10:90, v/o), thiab thaum kawg MeOH muab 250 feem (30 mL txhua). 85:15, thiab 80:20 raws li cov txheej txheem eluent (v/o), cov qauv zoo sib xws tau sib sau ua ke kom muab 16 feem (F1 txog F16).

Fraction F5 (181.2 mg) los ntawm thawj zaug CC eluted nrog hexane-CH, Cl2 (50: 50 txog 10: 90, v / o) raug rau ib qho tshiab silica gel CCusing tib txoj kev saum toj no eluted nrog hexane (100 feem pua), hexane CH, Cl2 (90: 10, 80: 20, 70: 30, 60: 40, 50: 50, 40: 60, 30: 70, 20: 80, thiab 10: 90, v / o), CH, Cl ,(100%) , CHCl2-EtOAc(90:10,80:20,70:30,60:40,50:50,40:60,30:70,20:80, thiab 10:90, thiab 10:90 , o/), EtOAc(100%), EtOAc-MeOH(85:15,70:30,50:50, 30:70, and 15:85, /o), thiab MeOH (100 feem pua) affording 118 feem( 15 mL txhua) uas tau muab sib sau ua ke hauv 12 ntu ntu (F5.1 txog F5.12), raws li TLC kev tshuaj xyuas kev soj ntsuam siv cov sib xyaw ntawm CH2Cl2-MeOH hauv qhov sib piv ntawm 95:05 thiab 90:10.Subfraction F5 .1 (28.9 mg) eluted nrog hexane (100 feem pua) raug rau kev npaj TLCeluted nrog CH, Cl-MeOH (95:05, v/v, ob elutions) them 2 (1.4 mg) thiab 3 (1.7 mg), raws. Subfractions F5.4 (21.5 mg) eluted nrog hexane-CH, C (80: 20, vo), F5.5 (16.7 mg) eluted nrog hexane-CH, Cl (80: 20.70: 30.60; 40. thiab 50: 50. , v /), thiab F5.6 (15.4 mg) eluted nrog hexane-CHCl2 (50: 50, 40: 60, thiab 30: 70, v / o) tau sib sau ua ke (53.6 mg) thiab tseem raug rau kev npaj TLC eluted nrog CH, Cl-MeOH (95:05, v/u, ob elutions) yielding 3 (8.9 mg).

Fraction F6(2099.1 mg) los ntawm thawj CC eluted nrog hexane-CHCl(10:9{229}},v/)), CH2Cl, (100 feem pua ​​), thiab CH, Cl2-EtOAc (90:10 txog 10:90, o/o) tau raug silica gel CC tshiab uas siv tib txoj kev qhia saum toj no eluted nrog tib cov kuab tshuaj systems uas them 140 feem (30 mL txhua). Tom qab TLCevaluation siv ib tug sib tov ntawm CH2Cl2-MeOH nyob rau hauv lub proportions ntawm 95:05, 90:10, thiab 85:15 raws li lub eluent system(o/o), cov qauv zoo sib xws tau muab sib sau ua ke muab 13 subfractions (F6 .1 to F6.13). Subfraction F6.6 (1541.3 mg) tau raug rau ib qho tshiab silica gel CC siv tib txoj kev saum toj no eluted nrog tib cov kuab tshuaj systems uas them 120 feem (30 mL txhua) uas tau soj ntsuam los ntawm TLC (siv tib txoj kev saum toj no) muab 12 tshiab subfractions (F6.6.1 txog F6.6.12). Subfrac-tion F6.6.2 (17.0 mg) eluted nrog hexane-CH, Cl, (60: 40 thiab 50: 50, v / v) raug rau kev npaj TLCeluted nrog CH2Cl2-MeOH(90: 10, v/o, ib elution) uas ua rau 3 (1.4mg).Subfraction F6.6.3(643.3 mg) eluted nrog hexane-CH2Cl2(50:50, 40:60, 30:70, 20:80, thiab 10:90, o/v), CH2Cl2 (100 feem pua), thiab CH2Cl2-EtOAc(90:10 thiab 80:20,o/o) tau raug tso tawm mus rau kev npaj TLCeluted nrog CH2Cl2-MeOH (90 : 10, vfo, ib qho elution) ua rau ib qho tshiab subfraction F6.6.3.1 (161.8 mg) uas tau raug rau qhov kev npaj tshiab TLC eluted nrog EtOAc-MeOH (90: 10, v / o, ib qho elution) affording 4(148.2 mg).Subfraction F6.6.4 (90.2 mg) eluted nrog CH2Cl2-EtOAc (70:30 thiab 60:40,o/o) raug rau prepara-tive TLCeluted nrog CH2Cl-MeOH (90:10, v/o, ib elution) yielding 1 (1.4 mg) thiab 4 (50.1 mg), feem. Subfraction F6.6.5(83.4 mg) eluted nrog CH, Cl-EtOAc(60:40 thiab 50:50, o/o) kuj raug rau kev npaj TLCeluted nrog CH, Cl2-MeOH(90:10, ufo, ib qho elution) them rov qab 1 (1.5 mg) thiab 4 (45.5 mg), feem. Subfraction F6.6.6 (101.4 mg) eluted nrog CH, Cl2-EtOAc (50: 50 thiab 40: 60, v/)) raug rau kev npaj TLC eluted nrog CH, Cl-MeOH (90: 10, vfo, ib qho elution) muab 8 (3.4 mg) thiab lwm qhov subfraction F6.6.6.1 (48.8 mg).Qhov no subfraction (F6.6.6.1) tau raug rau kev npaj TLCeluted pib nrog EtOAc-MeOH (85 :15, vo, ib elution) thiab posteriorly CHCl-MeOH(90:10,v/v, ib elution) rov tshwm sim hauv 4(37.4 mg). Rau qhov kev ua tiav no, lub phaj chromatographic tau pib eluted nrog lub sijhawm txawb EtOAc-MeOH (85: 15, v / o, ib qho elution). Tom qab qhov elution no, cov phaj chromatographic tau qhuav kom rov txav cov kuab tshuaj thiab tom qab ntawd raug rau qhov tshiab elution nrog rau theem mobile CH, Cl,-MeOH (90: 10, vf, ib elution) yielding 4.Subfraction F6.6.7 ( 109.0 mg)eluted nrog CH, Cl2-EtOAc(40:60,30:70,20:80 thiab 10:90,vfo) raug rau kev npaj TLC eluted nrog CH,Cl2-MeOH (90:10, vo, ib qho elution) muab lwm qhov subfraction F6.6.7.1 (24.8 mg) uas tau raug rau kev npaj TLC tshiab uas tau pib nrog EtOAc-MeOH (85: 15, v / o, ib qho elution) thiab posteriorly CH2Cl2-MeOH (90:10,vfo, ib qho elution) ua rau 9 (11.1 mg). Cov txheej txheem cais tawm siv rau 9 yog tib yam li tau piav qhia yav dhau los rau 4. Subfraction F6.6.10(92.0 mg) eluted nrog EtOAc-MeOH(60:40 thiab 50:50, v / o) raug rau kev npaj TLCeluted nrog CH, Cl,-MeOH (90:10, v/o, ib elution) yielding 5 (1.7 mg) thiab 6 (7.8 mg), feem. Subfraction F6.6.11 (326.0 mg) eluted nrog EtOAc-MeOH (50: 50, 40: 60, 30: 70, 20: 80, thiab 10: 90, v / o) raug rau kev npaj TLCeluted nrog CHCl- MeOH (90: 10, v / o, ib qho elution) vielding 1 (2.4 mg) thiab 7 (12.5 mg), feem.

Fraction F7 (692.4 mg) los ntawm thawj zaug CCeluted nrog EtOAc (100 feem pua) thiab CHCl2-EtOAc (95:05,90:10,85:15, thiab 75:25,ofo) tau raug silica gel tshiab CCusing tib txoj kev raws li tau piav qhia rau thawj zaug CC nrog tib cov kuab tshuaj sib xyaw ua ke uas them rau 110 feem (30 mL txhua). :15 raws li cov txheej txheem eluent (o / o), cov piv txwv zoo sib xws tau muab sib sau ua ke kom muab 12 qhov feem cuam tshuam (F7.1 rau F7.12).F7.8(147.4 mg) eluted nrog CH, Cl2- EtOAc (50: 50, 40: 60, thiab 30: 70, v / o feem pua) raug rau kev npaj TLC eluted nrog CHCl-MeOH (95: 05, vfo, ib elution) affording 10 (5.4 mg) thiab 11 ( 120.6 mg), feem. F7.9(144.2 mg) eluted nrog CHCl2-EtOAc (30:70, 20:80, thiab 10:90, v/o feem pua) kuj raug rau kev npaj TLCeluted nrog CH, Cl,-MeOH( 95:05, vo, ib elution) them 11 (91.2 mg) thiab sib tov ntawm 12 thiab 13 (39.4 mg), feem.

Thalifoline (1): Brown amorphous hmoov;' H-NMR thiab 13C-NMR raws li cov ntaub ntawv [32]; LR-ESI(plus)-MS [M plus H]* plus m/z 208.

Anonaine (2): Brown amorphous hmoov; 1H-NMR thiab 13C-NMR raws li cov ntaub ntawv [33]; LR-ESI(plus)-MS [M plus H] plus m/z 266.

Liriodenine (3): Yellow crystals (CH, Cl-MeOH 3: 1); 1H-NMR thiab 13C-NMR raws li cov ntaub ntawv [10,34]; LR-ESI(plus)-MS [M plus H] plus m/z 276.

(S)-(ntxiv )-Reticuline(4): Brown amorphous hmoov; [ ]p25 plus 71,60(c 0.05 g/100 mL, MeOH); 1H-NMR thiab 13C-NMR raws li cov ntaub ntawv [27;LR-ESI(plus)-MS [M ntxiv rau H] ntxiv rau m/z 30.

3. ): Brown amorphous hmoov; 'H thiab 13C-NMR cov ntaub ntawv, saib Table 1; LR-ESI(plus)-MS [M plus H] plus m/z 328.

(ntxiv rau ){{0}}S,2R-Reticuline N -oxide(6): Brown amorphous hmoov; lp25 plus 136.4 (c0.146g/100 mL, MeOH); 1Hand 13C-NMR cov ntaub ntawv, saib Table1;LR-APCI(ntxiv)-MS [M ntxiv rau H] ntxiv rau m/2346

(ntxiv ){{0}}S,2S-Reticuline N.-oxide (7): Brown amorphous hmoov; [ ]p25 ntxiv rau 394.5 (c 0.03g/100 mL, MeOH);'H thiab 13C-NMR cov ntaub ntawv, saib Table 1;LR-APCI(ntxiv )-MS [M ntxiv rau H] ntxiv m/z346.

Coreximine (8): Lub teeb daj amorphous hmoov;[]p25 ntxiv 201.1(c 0.08g/100mL, MeOH)'H-NMR thiab 13C-NMR raws li cov ntaub ntawv[34,35];LR- ESI(ntxiv)-MS [M ntxiv rau H] ntxiv rau m/z 328.

Bisnorargemonine (9): Brown amorphous hmoov; 1H-NMR thiab 13C-NMR raws li cov ntaub ntawv [36]; LR-APCI(ntxiv)-MS [M ntxiv rau H] ntxiv rau m/z 328.

Isochamanetin (10): Lub teeb daj amorphous hmoov; [ p25-12.4(c 0.5 g/100 mL, MeOH)'H-NMRand 13C-NMRin raws li cov ntaub ntawv[37;LR-APCI(-)-MS [MH]-m/z361.

Dichamanetin (11): Yellow amorphous hmoov;[ ]D25-10.7(c 0.5g/100 mL, MeOH); 1H-NMR thiab 13C-NMR raws li cov ntaub ntawv [37]; LR-ESI(-)-MS [MH]-m/z467.

Kev sib xyaw ntawm uvarinol (12) thiab isouvarinol (13): Yellow amorphous hmoov; H-NMR thiab 13C-NMR raws li cov ntaub ntawv [37]; LR-ESI(-)-MS [MH]- m/z 573.

3.4.In Vitro Cytotoxic Assay

3.4.1.Cells

HL-60 (tib neeg promyelocytic leukemia), MCF-7 (tib neeg lub mis adenocarcinoma), HepG2 (human hepatocellular carcinoma), HCT116 (tib neeg mob plab hnyuv), thiab MRC-5 (tib neeg lub ntsws fibroblast) Cov kab xov tooj ntawm tes tau txais los ntawm American Type Culture Collection (ATCC, Manassas, VA, USA), thiab tau coj los ua raws li kev pom zoo los ntawm ATCC tsiaj kab lis kev cai qhia. Tag nrho cov kab ntawm tes tau raug sim rau mycoplasma siv cov khoom siv mycoplasma stain (Sigma-Aldrich) kom siv tau cov hlwb tsis muaj kab mob.

3.4.2. Cytotoxicity Assay

Rau cytotoxicity assay, cell viability yog quantified los ntawm Alamar xiav txoj kev, raws li yav tas los piav [{{0}}]. Rau txhua qhov kev sim, cov hlwb tau muab tso rau hauv 96- cov phiaj zoo. Cov khoom xyaw tshuaj tau yaj hauv dimethyl sulfoxide (DMSO, Vetec Ouimica Fina Ltda., Duque de Caxias, RJ, Brazil) thiab ntxiv rau txhua qhov dej thiab incubated rau 72 teev. Doxorubicin (doxorubicin hydrochloride, purity Ntau dua lossis sib npaug li 95 feem pua, Laboratory IMA SAIC, Buenos Aires, Argentina) tau siv los ua kev tswj xyuas zoo. Thaum kawg ntawm kev kho mob, 20 μL ntawm cov tshuaj hauv Tshuag (0.312 mg / mL) ntawm resazurin (Sigma-Aldrich Co.) tau ntxiv rau txhua lub qhov dej Absorbances ntawm 570 nm thiab 600 nm tau ntsuas los ntawm SpectraMax190 Microplate Reader (Molecular Devices, Sunnyvale, CA, USA). nrog 95 feem pua ​​​​ntawm kev ntseeg siab (CI95 feem pua) siv software GraphPad Prism (Intuitive Software for Science; San Diego, CA, USA).

4. Cov lus xaus

Kev tshawb xyuas phytochemical ntawm cov tawv ntoo ntawm D.calycina coj mus rau kev sib cais thiab kev txheeb xyuas ntawm kaum peb lub tebchaw (1-13); cuaj isoquinoline-derived alkaloids (1-9) thiab plaubflavonoids(10-13). Cov alkaloids belongs rau cov subclasses: yooj yim isoquinoline, thali online (1); aporphine, anonaine (2); oxoaporphine, liriodenine (3); benzyl-tetrahydroisoquinolines, (ntxiv )-reticuline(4), de hydro-oxonorreticuline(3,4-dihydro{10}}hydroxy-6-}methoxy-1- isoquinolinyl)(3- }hydroxy-4-methoxyphenyl)-methanone)(5), 1S,2R-reticuline Ng-oxide(6), thiab 1S,2S-reticuline N.-oxide (7); tetrahydro protoberberine, coreximine (8); thiab pavine, bisnorargemonine (9). Thaum cov flavonoids yog benzylated dihydroflavones, isorhamnetin (10), dichamanetin (11), thiab sib tov ntawm uvarinol (12) thiab isouvarinol (13). Cov tshuaj sib cais tau piav qhia thawj zaug hauv D.calycina thiab hauv Diclinanona genus.

Kev ua haujlwm cytotoxic ntawm cov tshuaj sib cais (tshwj tsis yog rau anonaine, liriodenine, 1,2-dihydro-oxonorreticuline, thiab coreximine) raug soj ntsuam tiv thaiv qog noj ntshav (HL-60, MCF-7, HepG2, thiab HCT116) thiab non-cancerous(MRC-5) ​​kab cell. Ntawm lawv, cov txiaj ntsig tau zoo tshaj plaws tau pom rau benzylated dihydroflavones dichamanetin (10), thiab

Kev sib xyaw ntawm uvarinol (12) thiab isouvarinol (13) uas pom muaj kev ua haujlwm cytotoxic nruab nrab tiv thaiv kab mob qog noj ntshav kuaj (<20.0μg.ml-1）and low="" cytotoxicity="" against="" the="" non-cancerous="" cell="" line="" mrc-5(="">25. 0 ug·mL-'). Dichamanetin (11) tau qhia cytotoxic kev ua haujlwm tawm tsam HL-60 thiab HCT116 nrog IC50 qhov tseem ceeb ntawm 15.78ug·mL-1 (33.70 μmol-L-1) thiab 18.99 ug·mL-I (40.56 μmol) .L-1), raws li qhov sib xyaw ntawm uvarinol (12) thiab isouvarinol (13) tau pom cytotoxic kev ua haujlwm tawm tsam HL-60, nrog IC50 tus nqi ntawm 9.74 ug∶mL-I, thiab HCT116, nrog IC50 tus nqi ntawm 17.31 ug·mL-1. Cov txiaj ntsig ntawm cov kab mob qog no kuj tau piav qhia thawj zaug rau benzylated dihydroflavones. Cov kev ua cytotoxic ntawm reticuline, anonaine, thiab liriodenine tau tsim yav dhau los, uas liriodenine yog cov tshuaj muaj zog tshaj plaws.

Cov txiaj ntsig tau txais hauv txoj haujlwm no qhia tau hais tias cov tsiaj ntawm tsev neeg Annonaceae yog qhov muaj txiaj ntsig zoo ntawm cov tshuaj lom neeg lom neeg nrog cov khoom siv cytotoxic, thiab qhia txog kev txuas ntxiv ntawm lawv cov kev tshawb fawb hauv lwm cov qauv ntawm kev sim tshuaj lom neeg.

Cov ntaub ntawv

1. Couvreur, TLP; Pirie, MD; Chatrou, LW; Saunders, RMK; Su, YCF; Richardson, EJ; Erkens, RHJ Thaum ntxov evolutionary keeb kwm ntawm flowering nroj tsuag tsev neeg Annonaceae: Steady diversification thiab boreotropical geodispersal. J. Biogeogr. 2011, 38, 664–680. [CrossRef]

2. Corrêa, MP Dicionário das Plantas Úteis do Brasil e das Exóticas Cultivadas; IBDF Ministério da Agricultura: Rio de Janeiro, Brazil, 1984.

3. Leboeuf, M.; Cav, A.; Bhaumik, PK; Mukherjee, B.; Mukherjee, R. Lub phytochemistry ntawm Annonaceae. Phytochemistry 1982, 21, 2783–2813. [CrossRef]

4. Rupprecht, JK; Hui, Y.-H.; McLaughlin, JL Annonaceous acetogenins: Kev tshuaj xyuas. J. Nat. Prod. Xyoo 1990, 53, 237–278. [CrossRef]

5. Lúcio, ASSC; Almeida, JRGDS; Da-Cunha, EVL; Taures, JF; Filho, JMB Alkaloids ntawm Annonaceae: tshwm sim thiab muab tso ua ke ntawm lawv cov dej num lom neeg. Hauv Alkaloids; Knolker, H.-J., Ed.; Elsevier: Amsterdam, Lub Netherlands, 2015; Tshooj 5; Phau 74, p. 233–409. [CrossRef]

6. Souza, CAS; Nardelli, VB; Paz, WHP; Pinheiro, MLB; Rodrigues, ACBC; Bomfim, LM; Soares, MBP; Bezerra, DP; Chaw, JS; Koolen, HHF; ua al. Asarone-derived phenylpropanoids thiab isoquinoline-derived alkaloids los ntawm cov tawv ntoo ntawm Duguetia pycnastera (Annonaceae) thiab lawv cov cytotoxicities. Quim. Nov 2020, 43, 1397–1403. [CrossRef]

7. Araújo, MS; Silva, FMA; Koolen, HHF; Costa, EV Isoquinoline-derived alkaloids los ntawm cov tawv ntoo ntawm Guatteria olivacea (Annonaceae). Biochem. Syst. Ecol. 2020, 92, 104105. [CrossRef]

8. Ngouonpe, AW; Mbobda, ASW; Happi, GM; Mbiantcha, M.; Tau, OK; Li, MS; Lateef, M.; Tchouankeu, JC; Kouam, SF Natural khoom los ntawm cov nroj tsuag tshuaj Duguetia staudtii (Annonaceae). Biochem. Syst. Ecol. Xyoo 2019, 83, 22–25. [CrossRef]

9. Santos, RC; Souza, AV; Andrade-Silva, M.; Cruz, KCV; Kassuya, CAL; Cardoso, CAL; Vieira, MC; Formaggio, ASN Antioxidant, anti-rheumatic thiab anti-inflammatory kev tshawb fawb ntawm extract thiab dicentrinone los ntawm Duguetia furfuracea (A. St.-Hil.) Benth. & nuv. f. J. Ethnopharmacol. 2018, 211, 9–16. [CrossRef]

10. Costa, EV; Pinheiro, MLB; Sauza, ADL; Barison, UA; Campos, FR; Valdez, RH; Ueda-Nakamura, T.; Dias Filho, BP; Nakamura, CV Trypanocidal kev ua haujlwm ntawm oxoaporphine thiab pyrimidine- -carboline alkaloids los ntawm ceg ntawm Annona foetida Mart. (Annonaceae). Molecules 2011, 16, 9714–9720. [CrossRef]

11. Silva, IB; Tulli, ECO; Milita, GCG; Costa-Lotufo, LV; Pessoa, C.; Moraes, MO; Albuquerque, S.; Siqueira, JM Lub antitumoral, trypanocidal thiab antileishmanial kev ua ub no ntawm extract thiab alkaloids cais los ntawm Duguetia furacea. Phytomedicine 2009, 16, 1059–1063. [CrossRef] [PubMed]

12. Costa, EV; Pinheiro, MLB; Xavier, CM; Silva, JRA; Amaral, UA; Sauza, ADL; Barison, UA; Campos, FR; Ferreira, UA; Machado, GMC; ua al. Ib pyrimidine- -carboline thiab lwm yam alkaloids los ntawm Annona foetida nrog kev ua haujlwm antileishmanial. J. Nat. Prod. 2006, 69, 292–294. [CrossRef]