Biodistribution of methotrexate-monoclonal antibody conjugates and complexes: experimental and clinical studies

Pimm, M. V.; Clegg, J.A.; Caten, Jane E.; Ballantyne, KC; Perkins, Alan C.; Garnett, Martin C.; Baldwin, Robert

doi:10.1016/0305-7372(87)90039-9

Cited by 10 publications

(6 citation statements)

References 14 publications

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“…Furthermore, by use of agents able to inhibit lysosomal protein degradation (ammonium chloride (Ohkuma & Poole, 1978) and leupeptin (Seglen et al, 1979)), it was shown that, once internalised, the MTX-HSA-791T/36 conjugates require lysosomal degradation to release free MTX in order to exert their cytotoxic effect, also shown previously for such conjugates (Garnett et al, 1985). problems (Pimm et al, 1987) (Thorpe & Ross, 1982), radionuclides (Buchegger et al, 1988), enzymes (Bagshawe, 1987), and conventional cytotoxic drugs, such as vindesine , methotrexate (Embleton & Garnett, 1985), and daunomycin (Gallego et al, 1984 (Embleton & Garnett, 1985). A large proportion of this work has centred on the use of the anti-folate drug methotrexate (MTX) and the monoclonal antibody 791T/36, which was raised against the human osteogenic sarcoma cell line, 791T (Embleton et al, 1981), and recognises a 72,000 molecular weight glycoprotein (gp72) on the cell surface (Price et al, 1983 tance was overcome partially, suggesting that mechanisms of resistance other than transport deficiency may have been involved.…”

Section: Competitive Effect Offree Antibodymentioning

confidence: 63%

“…However, antibody-carrier-drug conjugates of this type have poor properties of biodistribution in vivo (Pimm et al, 1987), and poor tumour localisation and penetration due mainly to their larger size, and therefore the in vitro ability of these conjugates to overcome totally MTXtransport deficiency may not be realised in vivo.…”

Section: Competitive Effect Offree Antibodymentioning

confidence: 99%

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Monoclonal antibody targeting of methotrexate (MTX) against MTX-resistant tumour cell lines

Affleck

Embleton²

1992

Br J Cancer

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Section: Competitive Effect Offree Antibodymentioning

confidence: 63%

Section: Competitive Effect Offree Antibodymentioning

confidence: 99%

Monoclonal antibody targeting of methotrexate (MTX) against MTX-resistant tumour cell lines

Affleck

Embleton²

1992

Br J Cancer

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“…By flow cytometry, we showed that mAb3A33 antibody substituted with several MDP-polymer molecules were able to bind target cells as well as native mAb3A33 antibody and that the substitution of the monoclonal antibody with MDP-polymer molecules did not significantly impair the binding capacity of the antibody. Indirect conjugation of drugs to monoclonal antibodies via intermediate carriers have been reported to increase the amount of drug attached to antibody without impairing its antigen-binding activity: human serum albumin (27)(28)(29)(30)(31)(32)(33), dextran (34)(35)(36)(37)(38)(39), poly(L-glutamate acid) (40), a copolymer of N-(2-hydroxypropyl)methacrylamide containing an oligopeptide sequence (41), and succinylated polylysine (42) were used as intermediate carriers. In this paper, we used gluconoylated poly(L-lysine), a nonimmunogenic (43) and neutral polymer; the gluconoylation prevents a nonspecific binding to cells and increases the water solubility of poly(L-lysine) (11).…”

Section: Discussionmentioning

confidence: 99%

Activation of mouse macrophages by muramyl dipeptide coupled with an anti-macrophage monoclonal antibody

Midoux

Martin²,

Collet³

et al. 1992

Bioconjugate Chem.

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A rat IgG2a monoclonal antibody (mAb3A33) directed against the mouse Mac-1 antigen was conjugated with muramyl dipeptide (MDP) by using an intermediate polymer; under such conditions 75 MDP molecules were bound to one antibody molecule. A poly(L-lysine) polymer substituted with muramyl dipeptide and 3-(2-pyridyldithio)propionyl residues were prepared, the remaining lysine epsilon-amino groups were acylated with D-gluconolactone, leading to a neutral polymer; then a few polymer conjugates were coupled to mAb3A33 via a disulfide bridge. The binding capacity of the monoclonal antibody was preserved after conjugation with MDP-polymer molecules. Mouse peritoneal macrophages, incubated for 24 h with MDP-mAb3A33 conjugate became cytostatic against P815 mastocytoma cells, whereas unconjugated mAb3A33 and MDP-bound to a nonspecific rat IgG2a were ineffective. An enhancement of the cytostatic activity induced by MDP-mAb3A33 conjugate was obtained in the presence of gamma-IFN. These results show that several tens of MDP molecules can be linked to a macrophage-specific monoclonal antibody by using a neutral intermediate polymer without impairing the binding antibody capacity and that this type of MDP conjugate can efficiently activate macrophages and therefore could be the basis of the development of new antitumor therapy.

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“…The third option has been under development for some time in our laboratory where conjugates containing 30-40 molecules of drug to antibody have been produced, human serum albumin being used as a carrier. Preliminary biodistribution studies with these conjugates have shown rapid hepatic clearance and little tumour localization (Pimm et al, 1988). This clearance is probably due to a high net negative charge on the conjugate molecules, a problem not apparent with direct antibody conjugates of low substitution.…”

Section: Discussionmentioning

confidence: 99%

Biodistribution and tumour localization of a methotrexate‐monoclonal‐antibody 791T/36 conjugate in nude mice with human tumour xenografts

Pimm

Clegg

Garnett

et al. 1988

Intl Journal of Cancer

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The blood kinetics and tumour localization of a conjugate of methotrexate (MTX) and MAb 791T136 were examined i n nude mice with human tumour xenografts. The antibod moiety of the conjugate was detected by labelling with l2J and the drug moiety was assayed using a radioimmunoassay for methotrexate. After radioiodination, the drug moiety was co-precipitable with the radiolabel when TCA or rabbit antimouse IgG antiserum was used. Following i.v. injection, serum kinetics of both the antibody and the drug moieties of the conjugate were essentially similar, and the integrity of serumborne conjugate was confirmed by the co-precipitation of radiolabel and drug. The radiolabelled antibody moiety of the conjugate localized in tumour xenografts, with 5-7% of the injected dose being present per gram of tissue within 6 hr of injection, and the levels were maintained for up to 4 days. Analysis of tumour levels of the M T X moiety showed a progressive uptake over the 4-day observation period with up t o 4% of the injected dose being present per gram of tumour when the experiment was terminated. Parallel studies with free M T X showed rapid clearance from the blood and a maximum of 0.35% of the dose/g of tumour 30 min after injection.Control immunoglobulin conjugated t o M T X did not show tumour localization of either the antibody or the drug moieties. These studies confirm that in vivo M T X remains bound t o antibody in this type of drug antibody conjugate and demonstrate site-specific targeting of this therapeutic agent.The tumour localization of MAbs directed against human tumour-associated antigens has now been extensively documented both experimentally and clinically (Pimm, 1987). These observations have re-awakened interest in the use of antibodies as site-specific targeting agents for cancer therapy (Embleton, 1987;Baldwin and Byers, 1987). There are now reports of antibody conjugate synthesis with a number of widely used anti-tumor agents including anthracyclines (Pimm et al., 1982b;Gallego et al., 1984), anti-metabolites such as methotrexate (Garnett et al., 1983;Kulkarni et al., 1985; Kanellos et al., 1985), and vinca alkaloids (Rowland et al., 1985, 1986).Although there are reports of therapeutic effects with a number of these conjugates, at least in tumour xenograft systems (Kanellos ef al., Kulkarni et al., 1985;Rowland et al., 1985Rowland et al., , 1986Smyth et al., 1986) little attention has been paid to the pharmacokinetics of drugs conjugated to antibodies and, most importantly, there is little information on the comparative tumour levels of these drugs achieved after administration in free form or as conjugate. In the present studies the in vivo distribution of conjugates of methotrexate and the MAb (791T/ 36) has been examined in mice bearing human osteosarcoma xenografts. This has involved assessment of blood and tumour levels of both the drug and antibody moieties of the conjugate in comparison with drug linked to control immunoglobulin or given in free form. MATERIAL AND METHODS Monoclonal antibodyThe ...

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Biodistribution of methotrexate-monoclonal antibody conjugates and complexes: experimental and clinical studies

Cited by 10 publications

References 14 publications

Monoclonal antibody targeting of methotrexate (MTX) against MTX-resistant tumour cell lines

Monoclonal antibody targeting of methotrexate (MTX) against MTX-resistant tumour cell lines

Activation of mouse macrophages by muramyl dipeptide coupled with an anti-macrophage monoclonal antibody

Biodistribution and tumour localization of a methotrexate‐monoclonal‐antibody 791T/36 conjugate in nude mice with human tumour xenografts

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