Cell lines

MCF10A, MCF-7 and MDA-MB-231 human breast cancer cell lines

Preparation method

The solubility of this compound in DMSO is > 10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

50 μM; 24 h

Applications

In MCF10A, MCF-7 and MDA-MB-231 cells, Clotrimazole inhibited migration of MCF-7 and MDA-MB-231 cells by 32±5% and 59±6%, respectively, but had no effect on MCF10A cells. Clotrimazole inhibited mobility of MDA-MB-231 cells and MCF-7 cells. Also, clotrimazole reduced the viability of breast cancer cells.

Animal models

CAL27 xenograft mice model

Dosage form

150 mg/kg/body; 6 times a week for two weeks; intraperitoneally (i.p.)

Application

In CAL27 xenograft mice model, clotrimazole significantly decreased the tumor volume of CAL27 cell xenograft in nude mice by 57.9%. Compared with control mice, the mean weights of the excised tumors were approximately 53.6% lower in clotrimazole-treated mice. Clotrimazole increased the number of apoptotic tumor cells.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

Clotrimazole, bis-phenyl-(2-chlorophenyl)-1-imidazolyl methane, is a uniquely antifungal compound. In vitro, its spectrum includes dematiaceous, dermatophytes, dimorphic fungi and yeasts species. Its inhibitory concentrations in vitro were ≤ 4μg/mL for most susceptible fungi and ≤1 μg/mL for many species, particularly Trichophyton and Candida. Concentrations >20 μg/mL were fungicidal only [1]. It is also a specific Ca2+ activated K+ channel (Gardos channel) inhibitor [2]. Its IC50 to whole-cell currents in epithelial cells is 9 μmol/l [3].
Gardos channel contributes to pathologic water loss from erythrocytes and results in abnormal hemoglobin and promotes sickling in vitro. To avoid K+ and water loss via this channel was suggested as a potential therapy in vivo [2].
At concentrations ≤0.39 μg/mL, clotrimazole inhibited most isolates of C. neoformans, H. capsulatum and C. immitis. At concentrations < 0.78 μg/mL, clotrimazole did not inhibit one of C. neoformans. At 0.78 μg/mL, clotrimazole was fungicidal for all isolates, except a less susceptible isolate of H. capsulatum [1].
Subjects who had sickle cell anemia were treated with oral clotrimazole (10mg clotrimazole/kg/d for one week, and then daily dose was escalated by 10mg/kg each week). At a dosage of 20mg clotrimazole/kg/d, it was found that the Gardos channel was inhibited, cell K+ content was increased, erythrocyte dehydration was reduced, and hemoglobin levels was somewhat increased in all subjects. There are only three types of adverse effects, i.e. mild/moderate dysuria in all subjects and a reversible increase in plasma aspartic transaminase and alanine transaminase levels in two subjects treated with 30mg clotrimazole/kg/d [2].
References:
[1]. Smith Shadomy. In Vitro Antifungal Activity of Clotrimazole (Bay b 5097), Infection & Immunity. 1971, 4(2): 143-148.
[2]. Carlo Brugnara, Beatrice Gee, Carrie C. Armsby, et al. Therapy with Oral Clotrimazole Induces Inhibition of the Gardos Channel and Reduction of Erythrocyte Dehydration in Patients with Sickle Cell Disease. J. Clin. Invest., 1996, 97(5): 1227-1234.
[3]. Markus Bleich and Richard Warth. The very small-conductance K+ channel KVLQT1 and epithelial function. Pflügers Arch - Eur J Physiol, 2000, 440:202-206.