Summary.
A series of polyamine conjugates were synthesized and evaluated for their ability to target the polyamine transporter (PAT) in two Chinese hamster ovary (CHO) cell lines (PAT-active CHO and PAT-inactive CHOMG). This systematic study identified salient features of the polyamine architecture required to target and enter cells via the PAT. Indeed, the separation of charges, the degree of N-alkylation, and the spacer unit connecting the N1-terminus to the appended cytotoxic component (anthracene) were found to be key contributors to optimal delivery via the PAT. Using the CHO screen, the homospermidine motif (e.g., 4,4-triamine) was identified as a polyamine vector, which could enable the selective import of large N1-substituents (i.e., naphthylmethyl, anthracenylmethyl and pyrenylmethyl), which were cytotoxic to cells. The cell selectivity of this approach was demonstrated in B-16 murine melanoma cells and normal melanocytes (Mel-A). Three polyamine areas (recognition and transport, vesicle sequestration and polyamine-target interactions) were identified for future research.
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Abbreviations
- AZ:
-
Antizyme
- CHO:
-
chinese hamster ovary
- CHOMG:
-
polyamine transport deficient CHO cells
- DFMO:
-
α-difluoromethylornithine
- Mel-A:
-
normal melanocyte cell line
- MGBG:
-
methylglyoxalbis(guanylhydrazone)
- ODC:
-
ornithine decarboxylase
- PAT:
-
polyamine transporter
- SPD:
-
spermidine
References
SM Aziz M Yatin DR Worthen DW Lipke PA Crooks (1998) ArticleTitleA novel technique for visualising the intracellular localization and distribution of transported polyamines in cultured pulmonary artery smooth muscle cells J Pharm Biomed Anal 17 307–320 Occurrence Handle9638584 Occurrence Handle10.1016/S0731-7085(98)00016-8 Occurrence Handle1:CAS:528:DyaK1cXktVSltrc%3D
T Azzam H Eliyahu L Shapira M Linial Y Barenholz AJ Domb (2002) ArticleTitlePolysaccharide-oligoamine based conjugates for gene delivery J Med Chem 45 1817–1824 Occurrence Handle11960493 Occurrence Handle10.1021/jm0105528 Occurrence Handle1:CAS:528:DC%2BD38XitlOktL0%3D
M Belting S Persson L-A Fransson (1999) ArticleTitleProteoglycan involvement in polyamine uptake Biochem J 338 317–323 Occurrence Handle10024506 Occurrence Handle10.1042/0264-6021:3380317 Occurrence Handle1:CAS:528:DyaK1MXitVGks78%3D
M Belting K Mani M Jönsson F Cheng S Sandgren S Jonsson K Ding J-G Delcros L-A Fransson (2003) ArticleTitleGlypican-1 is a vehicle for polyamine uptake in mammalian cells: a pivotal role for nitrosothiol-derived nitric oxide J Biol Chem 278 47181–47189 Occurrence Handle12972423 Occurrence Handle10.1074/jbc.M308325200 Occurrence Handle1:CAS:528:DC%2BD3sXovFKktLc%3D
RJ Bergeron JS McManis CZ Liu Y Feng WR Weimar GR Luchetta Q Wu J Ortiz-Ocasio JRT Vinson D Kramer C Porter (1994) ArticleTitleAntiproliferative properties of polyamine analogues: a structure-activity study J Med Chem 37 3464–3476 Occurrence Handle7932575 Occurrence Handle10.1021/jm00047a004 Occurrence Handle1:CAS:528:DyaK2MXlsFSluw%3D%3D
RJ Bergeron JS McManis WR Weimar KM Schreier F Gao Q Wu J Ortiz-Ocasio GR Luchetta C Porter JRT Vinson (1995) ArticleTitleThe role of charge in polyamine analog recognition J Med Chem 38 2278–2285 Occurrence Handle7608892 Occurrence Handle10.1021/jm00013a003
RJ Bergeron Y Feng WR Weimar JS McManis H Dimova C Porter B Raisler O Phanstiel (1997) ArticleTitleA comparison of structure-activity relationships between spermidine and spermine analogue antineoplastics J Med Chem 40 1475–1494 Occurrence Handle9154970 Occurrence Handle10.1021/jm960849j Occurrence Handle1:STN:280:ByiB1Mbot1U%3D
RJ Bergeron R Müller J Bussenius JS McManis RL Merriman RE Smith H Yao WR Weimar (2000) ArticleTitleSynthesis and evaluation of hydroxylated polyamine analogues as antiproliferatives J Med Chem 43 224–235 Occurrence Handle10649978 Occurrence Handle10.1021/jm990375z Occurrence Handle1:CAS:528:DyaK1MXotFGgtbw%3D
RJ Bergeron JS McManis AM Franklin H Yao WR Weimar (2003) ArticleTitlePolyamine-Iron Chelator Conjugate J Med Chem 46 5478–5483 Occurrence Handle14640556 Occurrence Handle10.1021/jm0302694 Occurrence Handle1:CAS:528:DC%2BD3sXovVClt7s%3D
IS Blagbrough AJ Geall (1998) ArticleTitleHomologation of polyamines in the synthesis of lipo-spermine conjugates and related lipoplexes Tetrahedron Lett 39 443–446 Occurrence Handle10.1016/S0040-4039(97)10542-1
TL Byers R Wechter ME Nuttall AE Pegg (1989) ArticleTitleExpression of a human gene for polyamine transport in chinese hamster ovary cells Biochem J 263 745–752 Occurrence Handle2512913 Occurrence Handle1:CAS:528:DyaK3cXhvFGr
TL Byers AE Pegg (1990) ArticleTitleRegulation of polyamine transport in chinese hamster ovary cells J Cell Physiol 143 460–467 Occurrence Handle2113535 Occurrence Handle10.1002/jcp.1041430309 Occurrence Handle1:CAS:528:DyaK3cXltFarsbk%3D
RA Casero SuffixJr PM Woster (2001) ArticleTitleTerminally alkylated polyamine analogues as chemotherapeutic agents J Med Chem 44 1–26 Occurrence Handle11141084 Occurrence Handle10.1021/jm000084m Occurrence Handle1:CAS:528:DC%2BD3cXptVOmu78%3D
Cohen GM, Cullis P, Hartley JA, Mather A, Symons MCR, Wheelhouse RT (1992) Targeting of cytotoxic agents by polyamines: synthesis of a chloroambucil-spermidine conjugate. J Chem Soc Chem Commun 298–300
L Covassin M Desjardins R Charest-Gaudreault M Audette MJ Bonneau R Poulin (1999) ArticleTitleSynthesis of spermidine and norspermidine dimers as high affinity polyamine transport inhibitors Bioorg Med Chem Lett 9 1709–1714 Occurrence Handle10397506 Occurrence Handle10.1016/S0960-894X(99)00262-0 Occurrence Handle1:CAS:528:DyaK1MXktVCis74%3D
PM Cullis L Merson-Davies R Weaver (1995) ArticleTitleConjugation of a polyamine to the bifunctional alkylating agent chlorambucil does not alter the preferred cross linking site in duplex DNA J Am Chem Soc 117 8033–8034 Occurrence Handle10.1021/ja00135a029 Occurrence Handle1:CAS:528:DyaK2MXmvVegtr8%3D
PM Cullis RE Green L Merson-Davies N Travis (1999) ArticleTitleProbing the mechanism of transport and compartmentalisation of polyamines in mammalian cells Chem Biol 6 717–729 Occurrence Handle10508681 Occurrence Handle10.1016/S1074-5521(00)80019-8 Occurrence Handle1:CAS:528:DyaK1MXmvVOnsrc%3D
J-G Delcros S Tomasi S Carrington B Martin J Renault IS Blagbrough P Uriac (2002) ArticleTitleEffect of spermine conjugation on the cytotoxicity and cellular transport of acridine J Med Chem 45 5098–5111 Occurrence Handle12408721 Occurrence Handle10.1021/jm020843w Occurrence Handle1:CAS:528:DC%2BD38XnsVeksLY%3D
J-G Delcros S Tomasi S Duhieu M Foucault B Martin M Le Roch V Eifler-Lima J Renault P Uriac (2006) ArticleTitleEffect of polyamine homologation on the transport and biological properties of heterocyclic amidines J Med Chem 49 232–245 Occurrence Handle16392808 Occurrence Handle10.1021/jm050018q Occurrence Handle1:CAS:528:DC%2BD2MXht1KktrbO
RA Gardner J-G Delcros F Konate F Breitbeil SuffixIII B Martin M Sigman M Huang O Phanstiel SuffixIV (2004) ArticleTitleN1-substituent effects in the selective delivery of polyamine-conjugates into cells containing active polyamine transporters J Med Chem 47 6055–6069 Occurrence Handle15537360 Occurrence Handle10.1021/jm0497040 Occurrence Handle1:CAS:528:DC%2BD2cXos1Wjsrk%3D
RA Gardner M Belting K Svensson O Phanstiel SuffixIV (2007) ArticleTitleSynthesis and transfection efficiencies of new lipophilic polyamines J Med Chem 50 308–318 Occurrence Handle17228872 Occurrence Handle10.1021/jm0607101 Occurrence Handle1:CAS:528:DC%2BD28XhtlektL3N
M-P Hasne B Ullman (2005) ArticleTitleIdentification and characterization of a polyamine permease from the protozoan parasite Leishmania major J Biol Chem 280 15188–15194 Occurrence Handle15632173 Occurrence Handle10.1074/jbc.M411331200 Occurrence Handle1:CAS:528:DC%2BD2MXjtFSltrY%3D
WD Heston L Uly WR Fair DF Covey (1985) ArticleTitleCytotoxic activity of aziridinyl putrescine enhanced by polyamine depletion with alpha-difluoromethylornithine Biochem Pharmacol 34 2409–2410 Occurrence Handle3925954 Occurrence Handle10.1016/0006-2952(85)90806-8 Occurrence Handle1:CAS:528:DyaL2MXlsFentb8%3D
JL Holley A Mather RT Wheelhouse PM Cullis JA Hartley JP Bingham GM Cohen (1992a) ArticleTitleTargeting of tumor cells and DNA by a chlorambucil-spermidine conjugate Cancer Res 52 4190–4195 Occurrence Handle1:CAS:528:DyaK38XlvVCgtbo%3D
JL Holley A Mather PM Cullis MR Symons P Wardman RA Watt GM Cohen (1992b) ArticleTitleUptake and cytotoxicity of novel nitroimidazole-polyamine conjugates in Ehrlich ascites tumour cells Biochem Pharmacol 43 763–769 Occurrence Handle10.1016/0006-2952(92)90241-A Occurrence Handle1:CAS:528:DyaK38XisVWrs74%3D
M Huber JG Pelletier K Torossian P Dionne I Gamache R Charestgandreault M Audette R Poulin (1996) ArticleTitle2,2′-dithiobis(N-ethyl-spermine-5-carboxamide) is a high affinity, membrane-impermeant antagonist of the mammalian polyamine transport system J Biol Chem 271 27556–27563 Occurrence Handle8910341 Occurrence Handle10.1074/jbc.271.44.27556 Occurrence Handle1:CAS:528:DyaK28XmvVWmu78%3D
K Igarashi K Kashiwagi (1999) ArticleTitlePolyamine transport in bacteria and yeast Biochem J 344 633–642 Occurrence Handle10585849 Occurrence Handle10.1042/0264-6021:3440633 Occurrence Handle1:CAS:528:DC%2BD3cXislCgsw%3D%3D
V Kuksa R Buchan PKT Lin (2000) ArticleTitleSynthesis of polyamines, their derivatives, analogues and conjugates Synthesis 9 1189–1207 Occurrence Handle10.1055/s-2000-6405
OH Lowry NJ Rosenbraugh AL Farr RJ Randall (1951) ArticleTitleProtein measurement with the Folin Phenol reagent J Biol Chem 193 265–275 Occurrence Handle14907713 Occurrence Handle1:CAS:528:DyaG38XhsVyrsw%3D%3D
JL Mandel WF Flintoff (1978) ArticleTitleIsolation of mutant mammalian cells altered in polyamine transport J Cell Physiol 97 335–344 Occurrence Handle730773 Occurrence Handle10.1002/jcp.1040970308 Occurrence Handle1:CAS:528:DyaE1MXhvF2juw%3D%3D
B Martin F Possémé C Le Barbier F Carreaux B Carboni N Seiler J-P Moulinoux J-G Delcros (2001) ArticleTitleN-Benzylpolyamines as vectors of boron and fluorine for cancer therapy and imaging: synthesis and biological evaluation J Med Chem 44 3653–3664 Occurrence Handle11606130 Occurrence Handle10.1021/jm010897q Occurrence Handle1:CAS:528:DC%2BD3MXnsFOqur4%3D
B Martin F Possémé C Le Barbier F Carreaux B Carboni N Seiler J-P Moulinoux J-G Delcros (2002) ArticleTitle(Z)-1,4-diamino-2-butene as a vector of boron, fluorine, or iodine for cancer therapy and imaging: synthesis and biological evaluation Bioorg Med Chem 10 2863–2871 Occurrence Handle12110306 Occurrence Handle10.1016/S0968-0896(02)00147-5 Occurrence Handle1:CAS:528:DC%2BD38XltFahtLw%3D
LJ Marton VA Levin SJ Hervatin J Koch-Weser PP McCann A Sjoerdsma (1981) ArticleTitlePotentiation of the antitumor therapeutic effects of 1,3-bis(2-chloroethyl)-1-nitrosurea by alpha-difluoromethylornithine, an ornithine decarboxylase inhibitor Cancer Res 41 4426–4431 Occurrence Handle6796258 Occurrence Handle1:CAS:528:DyaL3MXmtFGiurk%3D
JLA Mitchell CL Simkus TK Thane P Tokarz MM Bonar B Frydman AL Valasinas VK Reddy LJ Marton (2004) ArticleTitleAntizyme induction mediates feedback limitation of the incorporation of specific polyamine analogues in tissue culture Biochem J 384 271–279 Occurrence Handle15315476 Occurrence Handle10.1042/BJ20040972 Occurrence Handle1:CAS:528:DC%2BD2cXhtVSqs7vM
O Phanstiel SuffixIV HL Price L Wang J Juusola M Kline SM Shah (2000) ArticleTitleThe effect of polyamine homologation on the transport and cytotoxicity properties of polyamine-(DNA-intercalator) conjugates J Org Chem 65 5590–5599 Occurrence Handle10970297 Occurrence Handle10.1021/jo0002792 Occurrence Handle1:CAS:528:DC%2BD3cXls1yit70%3D
C Porter J Miller RJ Bergeron (1984) ArticleTitleAliphatic chain-length specificity of the polyamine transport system in ascites L1210 leukemia cells Cancer Res 44 126–128 Occurrence Handle6690029 Occurrence Handle1:CAS:528:DyaL2cXps1Wlsg%3D%3D
CW Porter PF Cavanaugh B Ganis E Kelly RJ Bergeron (1985) ArticleTitleBiological properties of N4- and N1,N8-spermidine derivatives in cultured L1210 leukemia cells Cancer Res 45 2050–2057 Occurrence Handle3921235 Occurrence Handle1:CAS:528:DyaL2MXktlWqtL8%3D
N Seiler F Dezeure (1990) ArticleTitlePolyamine transport in mammalian cells Int J Biochem 22 211–218 Occurrence Handle2110083 Occurrence Handle10.1016/0020-711X(90)90332-W Occurrence Handle1:CAS:528:DyaK3cXhslGqtb4%3D
N Seiler J-G Delcros JP Moulinoux (1996) ArticleTitlePolyamine transport in mammalian cells. An update Int J Biochem Cell Biol 128 843–861 Occurrence Handle10.1016/1357-2725(96)00021-0
D Soulet L Covassin M Kaouass R Charest-Gaudreault M Audette R Poulin (2002) ArticleTitleRole of endocytosis in the internalisation of spermidine-C2-BODIPY, a highly fluorescent probe of polyamine transport Biochem J 367 347–357 Occurrence Handle12097141 Occurrence Handle10.1042/BJ20020764 Occurrence Handle1:CAS:528:DC%2BD38XotVWlsbs%3D
D Soulet B Gagnon S Rivest M Audette R Poulin (2004) ArticleTitleA fluorescent probe of polyamine transport accumulates into intracellular acidic vesicles via a two-step mechanism J Biol Chem 279 49355–49366 Occurrence Handle15208319 Occurrence Handle10.1074/jbc.M401287200 Occurrence Handle1:CAS:528:DC%2BD2cXpslKisbs%3D
PA Stark BD Thrall GG Meadows MM Abdul-Monem (1992) ArticleTitleSynthesis and evaluation of novel spermidine derivatives as targeted cancer chemotherapeutic agents J Med Chem 35 4264–4269 Occurrence Handle1447731 Occurrence Handle10.1021/jm00101a002 Occurrence Handle1:CAS:528:DyaK38XmtlCnurg%3D
A Valasinas A Sarkar VK Reddy LJ Marton HS Basu B Frydman (2001) ArticleTitleConformationally restricted analogues of 1N,14N-bisethylhomospermine (BE-4-4-4): synthesis and growth inhibitory effects on human prostate cancer cells J Med Chem 44 390–403 Occurrence Handle11462979 Occurrence Handle10.1021/jm000309t Occurrence Handle1:CAS:528:DC%2BD3cXovVWmsr4%3D
L Wang HL Price J Juusola M Kline O Phanstiel SuffixIV (2001) ArticleTitleThe Influence of polyamine architecture on the transport and topoisomerase II inhibitory properties of polyamine DNA-intercalator conjugates J Med Chem 44 3682–3691 Occurrence Handle11606133 Occurrence Handle10.1021/jm010181v Occurrence Handle1:CAS:528:DC%2BD3MXntVyntLk%3D
C Wang KA Abboud O Phanstiel SuffixIV (2002) ArticleTitleSynthesis and characterization of N1-(4-toluenesulfonyl)-N1-(9-anthracenemethyl)triamines J Org Chem 67 7865–7868 Occurrence Handle12398516 Occurrence Handle10.1021/jo020331p Occurrence Handle1:CAS:528:DC%2BD38Xnslamtbc%3D
C Wang J-G Delcros J Biggerstaff O Phanstiel SuffixIV (2003a) ArticleTitleSynthesis and biological evaluation of N1-(anthracen-9-ylmethyl)triamines as molecular recognition elements for the polyamine transporter J Med Chem 46 2663–2671 Occurrence Handle10.1021/jm030028w Occurrence Handle1:CAS:528:DC%2BD3sXjvVWrs7s%3D
C Wang J-G Delcros J Biggerstaff O Phanstiel SuffixIV (2003b) ArticleTitleMolecular requirements for targeting the polyamine transport system: synthesis and biological evaluation of polyamine-anthracene conjugates J Med Chem 46 2672–2682 Occurrence Handle10.1021/jm020598g Occurrence Handle1:CAS:528:DC%2BD3sXjvVahuro%3D
C Wang J-G Delcros L Cannon F Konate H Carias J Biggerstaff RA Gardner O Phanstiel SuffixIV (2003c) ArticleTitleDefining the molecular requirements for the selective delivery of polyamine-conjugates into cells containing active polyamine transporters J Med Chem 46 5129–5138 Occurrence Handle10.1021/jm030223a Occurrence Handle1:CAS:528:DC%2BD3sXot1Ojtr8%3D
ZM Yuan MJ Egorin DM Rosen MA Simon PS Callery (1994) ArticleTitleCellular pharmacology of N1- and N8-aziridinyl analogues of spermidine Cancer Res 54 742–748 Occurrence Handle8306336 Occurrence Handle1:CAS:528:DyaK2cXitlGmtLo%3D
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Phanstiel, O., Kaur, N. & Delcros, JG. Structure-activity investigations of polyamine-anthracene conjugates and their uptake via the polyamine transporter. Amino Acids 33, 305–313 (2007). https://doi.org/10.1007/s00726-007-0527-y
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DOI: https://doi.org/10.1007/s00726-007-0527-y