N-(2-Hydroxypropyl)methacrylamide (HPMA) copolymer carrier containing the anticancer drug doxorubicin bound either by a proteolytically degradable bond (non-targeted PK1 or targeted with alpha-CD71 mAb) or by a hydrolytically degradable bond were synthesised and tested in vivo for various biological properties. Mouse 38C13 B-cell lympoma was used as a well established and defined cell line for this study. 38C13 cells are sensitive to free doxorubicin and IC50 was very low, about 0.014 microM. PK1 showed a strongly decreased cytostatic effect, IC50 being 12.6 microM. alpha-CD71 targeted conjugate, which can be considered as an antibody-targeted form of PK1, had IC50 0.358 microM. HPMA copolymer with doxorubicin bound via a hydrolytically sensitive bond (HYD conjugate) showed a high cytostatic effect with IC50 about 0.052 microM. We demonstrated that HYD conjugate inhibited DNA synthesis and induced p21(Waf1/Cip1) protein expression (p21(Waf1/Cip1) is cyclin-dependent kinase inhibitor which blocks cell cycle progression) as quickly as free doxorubicin, whereas PK1 acted much more slowly. Similarly, apoptosis induction measured by Annexin V binding and Caspase 3 activity was detected later after incubation of cells with PK1 or alpha-CD71 targeted conjugate. Apoptosis was manifested by elevation of bax and bad mRNA levels, which was much more rapid and intense in the case of free doxorubicin and HYD conjugate. Expression of antiapoptotic genes as well as cyclin-dependent kinases was surprisingly not affected.
- MeSH
- akrylamidy chemická syntéza metabolismus farmakologie MeSH
- apoptóza účinky léků MeSH
- buněčná membrána účinky léků patologie MeSH
- buněčný cyklus genetika MeSH
- DNA antagonisté a inhibitory genetika metabolismus MeSH
- doxorubicin chemická syntéza metabolismus farmakologie MeSH
- exprese genu účinky léků genetika MeSH
- geny myc účinky léků genetika MeSH
- hydrazony chemická syntéza metabolismus farmakologie MeSH
- hydrolýza MeSH
- inhibiční koncentrace 50 MeSH
- inhibitor p21 cyklin-dependentní kinasy MeSH
- kaspasa 3 MeSH
- kaspasy škodlivé účinky účinky léků metabolismus MeSH
- ligandy * MeSH
- messenger RNA genetika MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- preklinické hodnocení léčiv metody MeSH
- proliferace buněk účinky léků MeSH
- protein Bad MeSH
- protein X asociovaný s bcl-2 MeSH
- proteiny buněčného cyklu genetika metabolismus MeSH
- protoonkogenní proteiny c-bcl-2 genetika metabolismus MeSH
- receptory transferinu účinky léků genetika MeSH
- thymidin metabolismus MeSH
- transportní proteiny genetika metabolismus MeSH
- tritium MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- akrylamidy MeSH
- Bad protein, mouse MeSH Prohlížeč
- Bax protein, mouse MeSH Prohlížeč
- Casp3 protein, mouse MeSH Prohlížeč
- Cdkn1a protein, mouse MeSH Prohlížeč
- DNA MeSH
- doxorubicin MeSH
- hydrazony MeSH
- inhibitor p21 cyklin-dependentní kinasy MeSH
- kaspasa 3 MeSH
- kaspasy MeSH
- ligandy * MeSH
- messenger RNA MeSH
- N-(2-hydroxypropyl)methacrylamide co-polymer-doxorubicin conjugate MeSH Prohlížeč
- protein Bad MeSH
- protein X asociovaný s bcl-2 MeSH
- proteiny buněčného cyklu MeSH
- protoonkogenní proteiny c-bcl-2 MeSH
- receptory transferinu MeSH
- thymidin MeSH
- transportní proteiny MeSH
- tritium MeSH
N-(2-Hydroxypropyl)methacrylamide (HPMA) copolymers containing the anticancer agent doxorubicin and targeted to the transferrin receptor either with anti-mouse CD71 monoclonal antibody (mAb) or with transferrin were synthesized to evaluate their binding and anti-proliferative activity in vitro and anti-tumor potential against 38C13 B-cell lymphoma in vivo. Both the doxorubicin and the targeting moieties were bound to HPMA copolymer chain by aminolysis via a Gly-Phe(D,L)-Leu-Gly spacer to ensure controlled intracellular release of the conjugated drug. We demonstrated that HPMA copolymer-bound doxorubicin targeted to the transferrin receptor with anti-mouse CD71 mAb strongly retards tumor growth, prolongs the survival and completely cures three out of nine experimental mice with established 38C13 tumors. The conjugate targeted with transferrin was less effective in vitro as well as in vivo. It completely cured only one out of seven experimental mice. Free or non-targeted HPMA copolymer-bound doxorubicin showed only a mild anti-tumor effect within the therapeutic schedule used. In vitro, HPMA copolymer-bound doxorubicin targeted with anti-mouse CD71 mAb shows approximately 4-fold higher cytotoxic effect than HPMA copolymer-bound doxorubicin targeted with transferin and 9-fold higher cytotoxic effect than non-targeted HPMA copolymer-bound doxorubicin.
- MeSH
- antibiotika antitumorózní aplikace a dávkování chemie farmakologie MeSH
- B-buněčný lymfom metabolismus MeSH
- biotin chemie MeSH
- buněčné dělení MeSH
- doxorubicin aplikace a dávkování chemie farmakologie MeSH
- indikátory a reagencie MeSH
- ligandy MeSH
- methakryláty chemie MeSH
- monoklonální protilátky aplikace a dávkování chemie MeSH
- myši inbrední C3H MeSH
- myši MeSH
- průtoková cytometrie MeSH
- receptory transferinu účinky léků metabolismus MeSH
- transferin aplikace a dávkování chemie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibiotika antitumorózní MeSH
- biotin MeSH
- doxorubicin MeSH
- hydroxypropyl methacrylate MeSH Prohlížeč
- indikátory a reagencie MeSH
- ligandy MeSH
- methakryláty MeSH
- monoklonální protilátky MeSH
- receptory transferinu MeSH
- transferin MeSH
Recombinant interferon alpha enhanced the MHC class I antigen density on human leukaemia/lymphoma cell lines REH, U-937 and HL-60, as measured by immunocytofluorometry using specific monoclonal antibodies. A similar effect was induced (as demonstrated in REH cells), also by human leukocyte interferon-alpha. The latter, however, caused no major alterations in the expression of leukocyte common antigen (ICA; CD45) and transferrin receptor (CD71) in the cell lines examined. In REH cells, there was no interferon-induced alteration of CD10 antigen (CALLA), which in this cell line is markedly down-regulated by 12-0-tetradecanoyl-phorbol-13-acetate (TPA). A decrease of CD4 antigen density on the cell membrane was induced by interferon-alpha in monoblastoid U-937 cells. No induction of MHC class I and II antigens by interferon-alpha was found in K-562 cell subline.
- MeSH
- akutní erytroblastická leukemie patologie MeSH
- akutní lymfatická leukemie patologie MeSH
- akutní promyelocytární leukemie patologie MeSH
- antigeny nádorové biosyntéza MeSH
- antigeny povrchové biosyntéza MeSH
- CD antigeny biosyntéza MeSH
- diferenciační antigeny biosyntéza MeSH
- difúzní velkobuněčný B-lymfom patologie MeSH
- interferon alfa-2 MeSH
- interferon alfa farmakologie MeSH
- leukocyty účinky léků imunologie patologie MeSH
- lidé MeSH
- nádorové buňky kultivované účinky léků imunologie MeSH
- neprilysin MeSH
- receptory transferinu biosyntéza účinky léků MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- rekombinantní proteiny MeSH
- tetradekanoylforbolacetát farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antigeny nádorové MeSH
- antigeny povrchové MeSH
- CD antigeny MeSH
- diferenciační antigeny MeSH
- interferon alfa-2 MeSH
- interferon alfa MeSH
- neprilysin MeSH
- receptory transferinu MeSH
- rekombinantní proteiny MeSH
- tetradekanoylforbolacetát MeSH
