Stability of Doxorubicin in Relation to Chemosensitivity Determinations: Loss of Lethality and Retention of Antiproliferative Activity

Abstract

The chemical stability of doxorubicin in a variety of tissue culture media has been studied by thin layer chromatography (TLC). In all the media examined, authentic doxorubicin was converted to a chemically distinct form as evidenced by the failure of this form to migrate on TLC plates. The rates of conversion were rapid enough (t 1/2 approximately equal to 3 hr) to be of consequence in chemosensitivity determinations, especially if working solutions of doxorubicin were to be routinely made and stored in tissue culture media. The addition of certain antioxidants to media did not prevent the conversion of authentic doxorubicin. However, doxorubicin was quite stable in distilled water. No single component of media was found to be responsible for the conversion of authentic doxorubicin, although arginine, histidine, tyrosine, NaHCO3, and Fe(NO3)3 could each generate a form of doxorubicin which did not migrate in TLC analysis. Purification of the nonmigrating form of doxorubicin demonstrated that in vitro conversion resulted in considerable loss of lethality while antiproliferative activity was retained. These observations provide possible explanations for the variability in chemosensitivity determinations and may explain some of the failures to predict clinical responsiveness.

Study of doxorubicin photodegradation in plasma, urine and cell culture medium by HPLC

“…The DXR metabolic pathway is now well documented (Israel e l al., 1978;Quattrone and Ranney, 1980) but nonmetabolic degradation, which converts anthracycline antibiotics in culture medium to 'pseudo'-metabolites, has not been well studied so far (Hoffman et al, 1979;Benvenuto et al, 1981;Robert, 1980;Ludwig and Alberto, 1984;Pavlik et al, 1984).…”

Novel Synthesized N-Ethyl-Piperazinyl-Amides of C2-Substituted Oleanonic and Ursonic Acids Exhibit Cytotoxic Effects through Apoptotic Cell Death Regulation

“…Compounds 4 and 6 were further subjected to DAPI (4 ,6-diamidino-2-phenylindole) staining in order to provide a more detailed picture of their antiproliferative and cytotoxic mechanisms. Prior to this assay, compound stability in cell culture media was evaluated after 72 h, using a thin-layer chromatography (TLC)-based protocol, with the necessary modifications [57,58]. Developed plates (using three different revealing agents) showed no other spots except the one corresponding to the original compound, suggesting that compounds 4 and 6 show no signs of decomposition in culture media for 72 h. Considering that compound 4 (highest overall cytotoxic activity) exhibited the highest overall cytotoxic activity on the melanoma cell line panel, the DAPI test was conducted on three human melanoma cell lines: A375, RPMI, and SK-MEL-28.…”

“…The stability of the assessed compounds in the culture media was evaluated by thin layer chromatography (TLC), this representing a viable and easy to achieve technique for this specific endpoint [57,58]. Compounds 4 and 6 were dissolved in DMSO, and the solutions were successively diluted to a 5 µM final concentration in culture media (DMEM and EMEM) so that the final DMSO concentration did not exceed 0.5%.…”

Degradation of Anthracycline Antitumor Compounds Catalysed by Metal Ions

“…Degradation of antitumor compounds is a common problem for pharmacologists. 1 The efficacy of an antitumor drug is related to the intracellular drug concentration as well as to the rate of cell uptake. 2 Anthracycline antibiotics, Doxorubicin (I) and Daunorubicin (l')(Chart 1) are benzanthraquinone drugs, which are useful in the treatement of several type of human malignancies.…”

“…Acta 1 9 9 3,11 77,236 1 2. Beraldo, H., Garnier-Suillerot, A., Tosi, L., and Lavelle, F. Biochemistry I 9 8 5,24,2841 3. Beraldo, H., Garnier-Suillerot, A., and Tosi, L. lnorg.…”