To avoid the side effects of the anti-cancer drug doxorubicin (Dox), we conjugated this drug to a N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer backbone. Dox was conjugated via an amide bond (Dox-HPMA(AM), PK1) or a hydrazone pH-sensitive bond (Dox-HPMA(HYD)). In contrast to Dox and Dox-HPMA(HYD), Dox-HPMA(AM) accumulates within the cell's intracellular membranes, including those of the Golgi complex and endoplasmic reticulum, both involved in protein glycosylation. Flow cytometry was used to determine lectin binding and cell death, immunoblot to characterize the presence of CD7, CD43, CD44, and CD45, and high-performance anion exchange chromatography with pulsed amperometric detector analysis for characterization of plasma membrane saccharide composition. Incubation of EL4 cells with Dox-HPMA(AM) conjugate, in contrast to Dox or Dox-HPMA(HYD), increased the amounts of membrane surface-associated glycoproteins, as well as saccharide moieties recognized by peanut agglutinin, Erythrina cristagalli, or galectin-1 lectins. Only Dox-HPMA(AM) increased expression of the highly glycosylated membrane glycoprotein CD43, while expression of others (CD7, CD44, and CD45) was unaffected. The binding sites for galectin-1 are present on CD43 molecule. Furthermore, we present that EL4 treated with Dox-HPMA(AM) possesses increased sensitivity to galectin-1-induced apoptosis. In this study, we demonstrate that Dox-HPMA(AM) treatment changes glycosylation of the EL4 T cell lymphoma surface and sensitizes the cells to galectin-1-induced apoptosis.

• MeSH
• Amides chemistry   MeSH
• Leukosialin metabolism   MeSH
• Apoptosis MeSH
• Doxorubicin analogs & derivatives   pharmacology   MeSH
• Endoplasmic Reticulum metabolism   MeSH
• Galectin 1 metabolism   MeSH
• Glycosylation MeSH
• Golgi Apparatus metabolism   MeSH
• Polymethacrylic Acids pharmacology   MeSH
• Lymphoma, T-Cell drug therapy   metabolism   pathology   MeSH
• Mice MeSH
• Cell Line, Tumor drug effects   MeSH
• Drug Carriers MeSH
• Cell Proliferation MeSH
• Antibiotics, Antineoplastic pharmacology   MeSH
• Flow Cytometry MeSH
• Blotting, Western MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Amides MeSH
• Leukosialin MeSH
• doxorubicin-N-(2-hydroxypropyl)methacrylamide copolymer conjugate MeSH Browser
• Doxorubicin MeSH
• Galectin 1 MeSH
• Polymethacrylic Acids MeSH
• Drug Carriers MeSH
• Antibiotics, Antineoplastic MeSH

Linkage of doxorubicin (Dox) to a water-soluble synthetic N-(2-hydroxypropyl)methacrylamide copolymer (PHPMA) eliminates most of the systemic toxicity of the free drug. In EL-4 lymphoma-bearing C57BL/6 mice, a complete regression of pre-established tumours has been achieved upon treatment with Dox-PHPMA-HuIg conjugate. The treatment was effective using a range of regimens and dosages, ranging from 62.5 to 100% cured mice treated with a single dose of 10-20 mg of Dox eq./kg, respectively. Fractionated dosages producing lower levels of the conjugate for a prolonged time period had substantial curative capacity as well. The cured mice developed anti-tumour protection as they rejected subsequently re-transplanted original tumour. The proportion of tumour-protected mice inversely reflected the effectiveness of the primary treatment. The treatment protocol leading to 50% of cured mice produced only protected mice, while no mice treated with early treatment regimen (i.e. starting on day 1 after tumour transplantation) rejected the re-transplanted tumour. Exposure of the host to the cancer cells was a prerequisite for developing protection. The anti-tumour memory was long lasting and specific against the original tumour, as the cured mice did not reject another syngeneic tumour, melanoma B16-F10. The immunity was transferable to naïve recipients in in vivo neutralization assay by spleen cells or CD8(+) lymphocytes derived from cured animals. We propose an effective treatment strategy which eradicates tumours without harming the protective immune anti-cancer responses.

• MeSH
• Doxorubicin analogs & derivatives   therapeutic use   MeSH
• Immunoglobulins therapeutic use   MeSH
• Immune Tolerance * MeSH
• Polymethacrylic Acids therapeutic use   MeSH
• Humans MeSH
• Lymphoma, T-Cell drug therapy   immunology   prevention & control   MeSH
• Melanoma, Experimental drug therapy   immunology   metabolism   MeSH
• Survival Rate MeSH
• Mice, Inbred C57BL MeSH
• Mice, Nude MeSH
• Mice MeSH
• Tumor Cells, Cultured transplantation   MeSH
• Skin Neoplasms drug therapy   immunology   metabolism   MeSH
• Drug Carriers MeSH
• Antibiotics, Antineoplastic therapeutic use   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• doxorubicin-N-(2-hydroxypropyl)methacrylamide copolymer conjugate MeSH Browser
• Doxorubicin MeSH
• Immunoglobulins MeSH
• Polymethacrylic Acids MeSH
• Drug Carriers MeSH
• Antibiotics, Antineoplastic MeSH

Allotransplantation (TPL) of the abdominal aortic segments of BN donors was performed in 32 Lewis recipients with or without cyclosporin A (CyA) immunosuppression, and the vascular changes were compared to those of 10 syngeneic grafts (Lewis-->Lewis) and to the autologous rat aortae. The vessels were examined 2, 3, 4 and 5 months post TPL by light microscopy, the thickness of intima and media was measured morphometrically and the cell infiltration of adventitia and intima was assessed semiquantitatively. Thirty-six aortae were examined by three-step enzyme immunohistochemistry (proof of selected differentiation, proliferation, cytoskeletal and connective tissue matrix antigens). The adventitia displayed an intense focal and scattered mononuclear cell infiltration; it was more discrete and focal in the intima. This cellularity persisted in the allografts but disappeared from the intima and was reduced in the adventitia of the isografts after four and five months. Disseminated ED1+ activated macrophages were the most prominent population of infiltrates whereas modest numbers of adventitial ED2+ tissue macrophages remained constant throughout the intervals examined. CD4+ cells (focal and scattered) outnumbered (roughly twice) the scattered CD8+ lymphocytes; both these types were rare in the intima. Leukocyte invasion of the media was lacking (except for scarce isolated CD8+ cells in some allografts). In syngeneic grafts the smooth muscle cells (SMC) of media remained intact and the intimal thickening was slight to absent (about 5 microns) four and five months post TPL. On the other hand, the allograft media underwent severe destructive changes (karyolysis, depletion of alpha-SMC actin, focal calcification and general thinning without rupture or aneurysm). The prominent allograft intimal thickening (70-80 microns) was due to the proliferation of longitudinally oriented myointimal cells (alpha-SMC actin, FD2, PCNA and Ki67+) and an increase in matrix substance (strong metachromasia and positivity of chondroitin-sulfate proteoglycan). The deposition of lipids remained discrete, without atheromatous plaques and mural thrombosis. All changes were comparable in CyA-treated and untreated animals. Thus the main lesions of the allografts were (i) persistent mononuclear infiltration chiefly in adventitia, (ii) destruction of medial SMC, and (iii) intimal thickening by proliferation of myointimal cells. At the postTPL intervals examined the proliferation and intimal migration of medial SMC were not apparent and a morphological correlate of significant anti-medial-SMC cytotoxic attack was lacking.

• MeSH
• Aorta, Abdominal pathology   transplantation   MeSH
• Transplantation, Homologous MeSH
• Immunohistochemistry MeSH
• Rats MeSH
• Rats, Inbred Lew MeSH
• Muscle, Smooth, Vascular pathology   MeSH
• Transplantation, Isogeneic MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Comparative Study MeSH

Drug targeting is an attractive new approach to killing cancer cells while leaving normal tissue unharmed. Recently we have developed a new generation of antibody-targeted immunosuppressive (cyclosporin A) and cytostatic (daunomycin, doxorubicin) drugs and photosensitizers (chlorin e6) effective in vitro and in vivo. The drugs and the targeting antibody (polyclonal and monoclonal) are conjugated to the oligopeptidic side chains of a water-soluble synthetic carrier, copolymer of N-(2-hydroxypropyl)methacrylamide. The composition of the side chains ensures the stability of the linkage between the drug and the polymeric carrier in the bloodstream and its intralysosomal degradability which is a prerequisite for the pharmacological activity of the preparation. Antibody-targeted polymer bound drugs show considerably decreased hepatotoxicity, cardiotoxicity, myelotoxicity and nephrotoxicity. Two adriamycin-HPMA copolymers are in Phase I/II clinical trials in United Kingdom.

• MeSH
• Acrylamides administration & dosage   adverse effects   chemistry   pharmacokinetics   MeSH
• Thy-1 Antigens immunology   MeSH
• Chlorophyllides MeSH
• Cyclosporine administration & dosage   adverse effects   pharmacokinetics   MeSH
• Daunorubicin therapeutic use   MeSH
• Doxorubicin analogs & derivatives   therapeutic use   MeSH
• Immunoconjugates * administration & dosage   adverse effects   chemistry   pharmacokinetics   MeSH
• Immunosuppressive Agents administration & dosage   adverse effects   chemistry   pharmacokinetics   MeSH
• Clinical Trials as Topic MeSH
• Polymethacrylic Acids therapeutic use   MeSH
• Humans MeSH
• Lysosomes metabolism   MeSH
• Histocompatibility Antigens Class II immunology   MeSH
• Antibodies, Monoclonal administration & dosage   chemistry   immunology   pharmacokinetics   MeSH
• Mice, Inbred BALB C MeSH
• Mice, Nude MeSH
• Mice MeSH
• Neoplasms drug therapy   MeSH
• Immune System Diseases drug therapy   MeSH
• Porphyrins administration & dosage   adverse effects   pharmacokinetics   MeSH
• Antibiotics, Antineoplastic administration & dosage   adverse effects   pharmacokinetics   MeSH
• Antineoplastic Agents administration & dosage   adverse effects   pharmacokinetics   MeSH
• Radiation-Sensitizing Agents administration & dosage   adverse effects   chemistry   pharmacokinetics   MeSH
• T-Lymphocyte Subsets immunology   MeSH
• Tissue Distribution MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Names of Substances
• Acrylamides MeSH
• Thy-1 Antigens MeSH
• Chlorophyllides MeSH
• Cyclosporine MeSH
• daunomycin-N-(2-hydroxypropyl)methacrylamide copolymer conjugate MeSH Browser
• Daunorubicin MeSH
• doxorubicin-N-(2-hydroxypropyl)methacrylamide copolymer conjugate MeSH Browser
• Doxorubicin MeSH
• Immunoconjugates * MeSH
• Immunosuppressive Agents MeSH
• Polymethacrylic Acids MeSH
• Histocompatibility Antigens Class II MeSH
• Antibodies, Monoclonal MeSH
• N-(2-hydroxypropyl)methacrylamide MeSH Browser
• phytochlorin MeSH Browser
• Porphyrins MeSH
• Antibiotics, Antineoplastic MeSH
• Antineoplastic Agents MeSH
• Radiation-Sensitizing Agents MeSH