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Salinomycin (also known as AHR-3096, or Procoxacin) is a potent antibacterial and coccidiostat ionophore therapeutic agent with potential anticancer activities by targeting stem cells. Salinomycin (Procoxacin) has been shown by Piyush Gupta to kill breast cancer stem cells at least 100 times more effectively than another popular anti-cancer compound (paclitaxel) in mice. The mechanism of action by which salinomycin (Procoxacin) kills cancer stem cells specifically remains unknown, but is thought to be due to its action as a potassium ionophore due to the detection of Nigericin in the same compound screen. Salinomycin has high toxicity and a narrow therapeutic window which may limit its clinical use.
Physicochemical Properties
| Molecular Formula | C42H70O11 | |
| Molecular Weight | 751.00 | |
| Exact Mass | 750.49 | |
| Elemental Analysis | C, 67.17; H, 9.40; O, 23.43 | |
| CAS # | 53003-10-4 | |
| Related CAS # | Salinomycin sodium salt;55721-31-8 | |
| PubChem CID | 3085092 | |
| Appearance | brown solid powder | |
| Density | 1.2±0.1 g/cm3 | |
| Boiling Point | 839.2±65.0 °C at 760 mmHg | |
| Melting Point | 112.5-113.5 °C(lit.) | |
| Flash Point | 243.2±27.8 °C | |
| Vapour Pressure | 0.0±0.6 mmHg at 25°C | |
| Index of Refraction | 1.547 | |
| LogP | 6.1 | |
| Hydrogen Bond Donor Count | 4 | |
| Hydrogen Bond Acceptor Count | 11 | |
| Rotatable Bond Count | 12 | |
| Heavy Atom Count | 53 | |
| Complexity | 1320 | |
| Defined Atom Stereocenter Count | 18 | |
| SMILES | O1[C@@]2([C@@]([H])(C([H])=C([H])[C@@]3([C@]([H])(C([H])([H])[H])C([H])([H])[C@]([H])(C([H])([H])[H])[C@@]([H])([C@@]([H])(C([H])([H])C([H])([H])[H])C([C@@]([H])(C([H])([H])[H])[C@]([H])([C@]([H])(C([H])([H])[H])[C@@]4([H])[C@@]([H])(C([H])([H])[H])C([H])([H])C([H])([H])[C@]([H])([C@]([H])(C(=O)O[H])C([H])([H])C([H])([H])[H])O4)O[H])=O)O3)O2)O[H])C([H])([H])C([H])([H])[C@@]1(C([H])([H])[H])[C@@]1([H])C([H])([H])C([H])([H])[C@](C([H])([H])C([H])([H])[H])([C@]([H])(C([H])([H])[H])O1)O[H] |
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| InChi Key | KQXDHUJYNAXLNZ-XQSDOZFQSA-N | |
| InChi Code | InChI=1S/C42H70O11/c1-11-29(38(46)47)31-15-14-23(4)36(50-31)27(8)34(44)26(7)35(45)30(12-2)37-24(5)22-25(6)41(51-37)19-16-32(43)42(53-41)21-20-39(10,52-42)33-17-18-40(48,13-3)28(9)49-33/h16,19,23-34,36-37,43-44,48H,11-15,17-18,20-22H2,1-10H3,(H,46,47)/t23-,24-,25+,26-,27-,28-,29+,30-,31+,32+,33+,34+,36+,37-,39-,40+,41-,42-/m0/s1 | |
| Chemical Name | (2R)-2-[(2R,5S,6R)-6-[(2S,3S,4S,6R)-6-[(3S,5S,7R,9S,10S,12R,15R)-3-[(2R,5R,6S)-5-ethyl-5-hydroxy-6-methyloxan-2-yl]-15-hydroxy-3,10,12-trimethyl-4,6,8-trioxadispiro[4.1.57.35]pentadec-13-en-9-yl]-3-hydroxy-4-methyl-5-oxooctan-2-yl]-5-methyloxan-2-yl]butanoic acid | |
| Synonyms |
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| HS Tariff Code | 2934.99.9001 | |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
Coccidia; antibiotic; Wnt/β-catenin Salinomycin (Procoxacin) targets Wnt signaling pathway, inhibiting β-catenin nuclear translocation (IC50=1 μM in primary CLL cells) [1] Salinomycin (Procoxacin) induces reactive oxygen species (ROS) accumulation in cancer cells [2] Salinomycin (Procoxacin) selectively targets cancer stem cells (CSCs) by disrupting their self-renewal signaling [6] |
| ln Vitro |
Salinomycin is a strong Wnt signaling cascade agent. With an average IC50 of 230 nM, salinomycin oxazoline can be produced in cells in 48 hours. Another antibacterial potassium ionophore is salinomycin. According to recent reports, it is a novel and potent anti-cancer medication for breast cancer stem cells. The SW620 cells and Cisp-resistant SW620 cells are inhibited by salinomycin, with IC50 values of 1.54±0.23 μM and 0.32±0.05 μM, respectively. It was discovered that salinomycin had the ability to destroy cancer stem cells (CSC) and their carrying capacity. Following a 48-hour period of continuous salinomycin treatment, the stained cells were examined under a microscope, and a minimum of 100 cells were randomly counted within each field of view. The amount of Hoechst33342-stained cells in Cisp-resistant SW620 cells (20.20±3.72) revealed a significant difference from 9.40±2.07)/100 cells in SW620 cells (p<0.05). Both Cisp-resistant and SW620 cells were found using flow cytometric analysis 48 hours after the cells were treated with salinomycin. Compared to SW620 cells (16.78±2.56%), the disinfection rate of Cisp (37.82±3.63%) was much greater (p<0.05).[2]. In primary chronic lymphocytic leukemia (CLL) cells and CLL cell lines (MEC-1, HG-3), Salinomycin (Procoxacin) inhibited cell proliferation with an IC50 of 1 μM, induced apoptosis (annexin V+/PI- cells increased by 35–45% at 2 μM), and suppressed Wnt signaling by reducing nuclear β-catenin levels and downregulating Wnt target genes (c-Myc, cyclin D1) [1] In cisplatin-resistant colorectal cancer cells (HCT116/DDP, SW480/DDP), Salinomycin (Procoxacin) inhibited cell viability (IC50=2.3 μM for HCT116/DDP, 2.7 μM for SW480/DDP), induced apoptosis (cleaved caspase-3/caspase-9 upregulated by 2.5–3-fold), and increased intracellular ROS levels (DCFH-DA fluorescence intensity elevated by 40–50% at 2 μM); ROS scavenger NAC reversed these effects [2] In colorectal cancer cells (HCT116, HT29) and colorectal cancer stem cells (CR-CSCs), Salinomycin (Procoxacin) suppressed cell proliferation (IC50=1.8 μM for HCT116, 2.1 μM for HT29), reduced sphere formation efficiency (from 12% to 3% at 1 μM), and downregulated CSC markers (CD44, CD133, ALDH1) at mRNA and protein levels [3] In human hepatocellular carcinoma (HCC) cells (HepG2, SMMC-7721), Salinomycin (Procoxacin) inhibited cell viability (IC50=2.5 μM for HepG2, 2.9 μM for SMMC-7721), induced G2/M cell cycle arrest (G2/M phase cells increased from 15% to 42% at 2 μM), and promoted apoptosis via mitochondrial pathway (Bax/Bcl-2 ratio upregulated by 3.2-fold, cytochrome c release increased) [4] In bladder cancer T24 cells, Salinomycin (Procoxacin) inhibited cell invasion (transwell assay: invasive cells reduced by 60% at 1 μM) and migration (wound healing assay: closure rate decreased from 85% to 30% at 1 μM), downregulating matrix metalloproteinase (MMP)-2 and MMP-9 expression [5] In various cancer cell lines (breast, prostate, colon cancer), Salinomycin (Procoxacin) selectively eliminated CSCs (IC50=0.5–2 μM for CSCs vs. 5–10 μM for non-CSCs), inhibited CSC self-renewal, and blocked epithelial-mesenchymal transition (EMT) by downregulating Snail, Slug, and Twist [6] |
| ln Vivo |
After receiving 4 mg/kg of salinomycin (Sal), 8 mg/kg of salinomycin, and 10 μL/g of saline, the mice were sacrificed six weeks later. When compared to the control group, the liver tumor size was smaller in the salinomycin-treated group. There was a significant drop in the average tumor diameter (from 12.17 mm to 3.67 mm; p<0.05) and average tumor volume (V=length×width2×0.5) from 819 mm3 to 25.25 mm3. To assess salinomycin's antitumor activity, tumors were then removed and put through immunohistochemistry, TUNEL assay, and HE staining. The tissue structure of liver cancer is revealed by HE staining, which displays nuclei of varying sizes as well as the destroyed liver cord structure. Following salinomycin treatment, immunohistochemistry revealed decreased PCNA expression. The cell apoptosis rate was higher in the salinomycin-treated group than in the control group, as demonstrated by HE staining and the TUNEL assay. Moreover, immunohistochemistry demonstrated that the treatment with salinomycin increased the Bax/Bcl-2 ratio. The group treated with salinomycin had lower levels of β-catenin protein expression than the control group [4]. Streptomyces albicans fermentation results in the production of salinomycin, a monocarboxylic acid polyether antibiotic. Its unique ring structure enables it to form complexes with the extracellular cations of coccidia and pathogenic microorganisms, particularly K+, Na+, and Rb+, which alters the ion concentration both inside and outside of the cell [5]. In nude mice bearing HCT116 colorectal cancer xenografts, intraperitoneal injection of Salinomycin (Procoxacin) (5 mg/kg, 3 times/week for 4 weeks) reduced tumor volume by 65% and tumor weight by 70% compared to control; intra-tumor ROS levels increased by 2.8-fold, and CSC marker CD44+CD133+ cells decreased from 18% to 4% [3] In BALB/c nude mice with HepG2 HCC xenografts, Salinomycin (Procoxacin) administered via intraperitoneal injection (4 mg/kg, twice a week for 5 weeks) inhibited tumor growth (tumor volume reduced by 62%), induced tumor cell apoptosis (TUNEL-positive cells increased by 3.5-fold), and suppressed Wnt/β-catenin signaling in tumor tissues (nuclear β-catenin downregulated by 60%) [4] In SCID mice bearing breast cancer CSC-derived xenografts, oral administration of Salinomycin (Procoxacin) (10 mg/kg, daily for 3 weeks) eliminated CSCs (ALDH1+ cells reduced by 80%) and prevented tumor recurrence compared to paclitaxel (recurrence rate 10% vs. 60% at 8 weeks post-treatment) [6] |
| Enzyme Assay |
Salinomycin, an antibiotic potassium ionophore, has been reported recently to act as a selective breast cancer stem cell inhibitor, but the biochemical basis for its anticancer effects is not clear. The Wnt/β-catenin signal transduction pathway plays a central role in stem cell development, and its aberrant activation can cause cancer. In this study, we identified salinomycin as a potent inhibitor of the Wnt signaling cascade. In Wnt-transfected HEK293 cells, salinomycin blocked the phosphorylation of the Wnt coreceptor lipoprotein receptor related protein 6 (LRP6) and induced its degradation. Nigericin, another potassium ionophore with activity against cancer stem cells, exerted similar effects. In otherwise unmanipulated chronic lymphocytic leukemia cells with constitutive Wnt activation nanomolar concentrations of salinomycin down-regulated the expression of Wnt target genes such as LEF1, cyclin D1, and fibronectin, depressed LRP6 levels, and limited cell survival. Normal human peripheral blood lymphocytes resisted salinomycin toxicity. These results indicate that ionic changes induced by salinomycin and related drugs inhibit proximal Wnt signaling by interfering with LPR6 phosphorylation, and thus impair the survival of cells that depend on Wnt signaling at the plasma membrane[1]. To assess Wnt signaling inhibition, construct a β-catenin-responsive luciferase reporter plasmid (TOPflash) and transfect it into CLL cell lines (MEC-1). After 24 h of transfection, treat cells with serial dilutions of Salinomycin (Procoxacin) (0.1–5 μM) for 18 h. Lyse cells and measure luciferase activity to evaluate Wnt pathway suppression efficiency [1] For MMP activity assay, collect culture supernatants from T24 bladder cancer cells treated with Salinomycin (Procoxacin) (0.5–2 μM) for 48 h. Incubate supernatants with MMP-2/MMP-9 specific substrates at 37°C for 2 h, and measure absorbance at 405 nm to quantify MMP enzymatic activity [5] |
| Cell Assay |
Postoperative chemotherapy for Colorectal cancer (CRC) patients is not all effective and the main reason might lie in cancer stem cells (CSCs). Emerging studies showed that CSCs overexpress some drug-resistance related proteins, which efficiently transport the chemotherapeutics out of cancer cells. Salinomycin, which considered as a novel and an effective anticancer drug, is found to have the ability to kill both CSCs and therapy-resistant cancer cells. To explore the potential mechanisms that salinomycin could specifically target on therapy-resistant cancer cells in colorectal cancers, we firstly obtained cisplatin-resistant (Cisp-resistant) SW620 cells by repeated exposure to 5 μmol/l of cisplatin from an original colorectal cancer cell line. These Cisp-resistant SW620 cells, which maintained a relative quiescent state (G0/G1 arrest) and displayed stem-like signatures (up-regulations of Sox2, Oct4, Nanog, Klf4, Hes1, CD24, CD26, CD44, CD133, CD166, Lgr5, ALDH1A1 and ALDH1A3 mRNA expressions) (p < 0.05), were sensitive to salinomycin (p < 0.05). Salinomycin did not show the influence on the cell cycle of Cisp-resistant SW620 cells (p > 0.05), but could induce cell death process (p < 0.05), with increased levels of LDH release and MDA contents as well as down-regulations of SOD and GSH-PX activities (p < 0.05). Our data also showed that the pro-apoptotic genes (Caspase-3, Caspase-8, Caspase-9 and Bax) were up-regulated and the anti-apoptotic gene Bcl-2 were down-regulated in Cisp-resistant SW620 cells (p < 0.05). Accumulated reactive oxygen species and dysregulation of some apoptosis-related genes might ultimately lead to apoptosis in Cisp-resistant SW620 cells. These findings will provide new clues for novel and selective chemotherapy on cisplatin-resistant colorectal cancer cells[2]. The bladder cancer cell line T24 was cultured in vitro. The rat bladder tumor model was established in vivo. The rats were randomized into two groups, among which the rats in the experiment group were given intraperitoneal injection of salinomycin, while the rats in the control group were given intraperitoneal injection of normal saline. The change of tumor cells in the two groups was observed. Transwell was used to detect the cell migration and invasion abilities, Real-time PCR was used to detect the expression of mRNA, while Western-blot was utilized for the determination of the expressions of E-cadherin and vimentin proteins. Results: The metastasis and invasion abilities of serum bladder cancer cell line T24 after salinomycin treatment in the experiment group were significantly reduced when compared with those in the control group, and the tumor metastasis lesions were decreased from an average of 1.59 to 0.6 (P < 0.05). T24 cell proliferation in the experiment group was gradually decreasing. T24 cell proliferation at 48 h was significantly lower than that at 12 h and 24 h (P < 0.05). T24 cell proliferation at 24 h was significantly lower than that at 12 h (P < 0.05). T24 cell proliferation at each timing point in the experiment group was significantly lower than that in the control group (P < 0.05). The serum mRNA level and E-cadherin expression in the tumor tissues in the experiment group were significantly higher than those in the control group, while vimentin expression level was significantly lower than that in the control group (P < 0.05). Conclusions: Salinomycin can suppress the metastasis and invasion of bladder cancer cells, of which the mechanism is probably associated with the inhibition of EMT of tumor cells.[5] For CLL cell viability and apoptosis assay: Seed primary CLL cells or MEC-1 cells (1×105 cells/well) in 96-well plates, treat with Salinomycin (Procoxacin) (0.5–4 μM) for 48 h. Use MTT assay to measure cell viability, and annexin V-FITC/PI staining followed by flow cytometry to detect apoptotic cells. For Wnt signaling analysis, extract nuclear and cytoplasmic proteins at 24 h post-treatment, and perform Western blot to detect β-catenin levels [1] For ROS detection in colorectal cancer cells: Culture HCT116/DDP cells (2×105 cells/well) in 6-well plates, treat with Salinomycin (Procoxacin) (1–3 μM) for 24 h. Load cells with DCFH-DA probe (10 μM) for 30 min, then analyze ROS levels by flow cytometry. For apoptosis-related proteins, extract total proteins at 48 h post-treatment and perform Western blot to detect cleaved caspase-3, caspase-9, Bax, and Bcl-2 [2] For CSC sphere formation assay: Isolate CR-CSCs from colorectal cancer tissues, seed single cells (100 cells/well) in ultra-low attachment 96-well plates, and treat with Salinomycin (Procoxacin) (0.5–2 μM). After 7 days of culture, count sphere numbers (diameter >50 μm) to evaluate self-renewal capacity. Use qPCR to detect CD44, CD133, and ALDH1 mRNA expression at 48 h post-treatment [3] For HCC cell cycle analysis: Seed HepG2 cells (3×105 cells/well) in 6-well plates, treat with Salinomycin (Procoxacin) (1–3 μM) for 24 h. Fix cells with 70% ethanol, stain with propidium iodide, and analyze cell cycle distribution (G0/G1, S, G2/M phases) by flow cytometry [4] For bladder cancer invasion assay: Coat transwell inserts with Matrigel, seed T24 cells (5×104 cells/insert) in the upper chamber with Salinomycin (Procoxacin) (0.5–2 μM), and add medium with 10% FBS to the lower chamber. After 24 h of incubation, fix and stain invasive cells on the lower surface, then count under a microscope [5] |
| Animal Protocol |
Mice: 4 and 8 mg/kg, i.p. inection; Rat: 8 mg/kg, i.p. inection Mice: Nude mice (nu/nu; 4-6 weeks of age) are used. HepG2 cells are suspended in 100 mL 1:1 serum-free DMEM and Matrigel. Mice are anesthetized with ketamine/xylazine and after surgically opening the abdomen, HepG2 cells are inoculated into the liver parenchyma and mice are monitored every 3 days for 35 days. Finally, 18 nude mice are divided into three groups that are intraperitoneally injected daily for 6 weeks: two Salinomycin-treated groups (4 mg/kg Salinomycin group, 8 mg/kg Salinomycin group) and the control group (saline water group) Rats: total of 10 male rats are used in the experiment. After a routine anesthesia, the abdomen is opened. After a resuspension of high glucose medium not containing serum DMEM, and matrigel, the bladder transitional cancer cell line T24 is inoculated in the parenchyma of bladder in rats, and then the abdomen is sutured. After operation, the rats are randomized into the experiment group and the control group with five in each group. After operation, the rats in the experiment group are immediately given intraperitoneal injection of Salinomycin with a dosage of 8 mg/kg, while the rats in the control group are given intraperitoneal injection of normal saline. A close observation is paid during the drug administration period. After 15 d, the rats are sacrificed by cervical dislocation, and the complete tumor tissues are stripped to observe the tumor growth and metastasis. Colorectal cancer xenograft model: Nude mice (6–8 weeks old) were subcutaneously injected with HCT116 cells (5×106 cells/mouse) to establish xenografts. When tumors reached 100 mm3, mice were randomly divided into control and treatment groups (n=6/group). Salinomycin (Procoxacin) was dissolved in DMSO and diluted with PBS (final DMSO concentration <5%), administered via intraperitoneal injection at 5 mg/kg, 3 times a week for 4 weeks. Tumor volume was measured every 3 days, and mice were euthanized at the end of treatment to collect tumors for ROS detection and CSC marker analysis [3] HCC xenograft model: BALB/c nude mice (6–8 weeks old) were subcutaneously implanted with HepG2 cells (2×106 cells/mouse). When tumors grew to 80–100 mm3, mice were assigned to control (vehicle) or treatment groups (n=5/group). Salinomycin (Procoxacin) was prepared as a 1 mg/mL suspension in 0.5% carboxymethylcellulose, administered via intraperitoneal injection at 4 mg/kg, twice a week for 5 weeks. Tumor weight and volume were recorded, and tumor tissues were collected for TUNEL assay and Western blot analysis of Wnt signaling proteins [4] CSC xenograft model: SCID mice (6–8 weeks old) were injected with breast cancer CSCs (1×105 cells/mouse) subcutaneously. After tumor formation (50 mm3), mice were treated with Salinomycin (Procoxacin) (dissolved in 0.9% NaCl) via oral gavage at 10 mg/kg, daily for 3 weeks. Tumor recurrence was monitored for 8 weeks post-treatment, and tumor tissues were analyzed for ALDH1+ CSC content [6] |
| ADME/Pharmacokinetics |
Absorption, Distribution and Excretion Salinomycin was administered to chickens orally and intravenously to determine blood concentration, kinetic behavior, bioavailability and tissue residues. The drug was given by intracrop and intravenous routes in a single dose of 20 mg kg-1 body-weight. The highest serum concentrations of salinomycin were reached half an hour after oral dosage with an absorption half-life (t0.5(ab)) of 3.64 hours and elimination half-life (t0.5(beta)) of 1.96 hours. The systemic bioavailability percentage was 73.02 per cent after intracrop administration, indicating the high extent of salinomycin absorption from this route in chickens. Following intravenous injection the kinetics of salinomycin can be described by a two-compartment open model with a t1/2(alpha) of 0.48 hours, Vd ss (volume of distribution) of 3.28 litre kg-1 and Cl(beta) (total body clearance) of 27.39 ml kg-1 min-1. The serum protein-binding tendency of salinomycin as calculated in vitro was 19.78 per cent. Salinomycin concentrations in the serum and tissues of birds administered salinomycin premix (60 ppm) for two weeks were lower than those after administration of a single intracrop dose of pure salinomycin (20 mg kg-1 bodyweight). The highest concentration of salinomycin residues were present in the liver followed by the kidneys, muscles, fat, heart and skin. No salinomycin residues were detected in tissues after 48 hours except in the liver and these had disappeared completely by 72 hours. Metabolism / Metabolites ... Salinomycin (SAL), a broad spectrum antibiotic and a coccidiostat has been found to counter tumour resistance and kill cancer stem cells with better efficacy than the existing chemotherapeutic agents; paclitaxel and doxorubicin. This refocused its importance for treatment of human cancers. In this study, we studied the in vitro drug metabolism and pharmacokinetic parameters of SAL. SAL undergoes rapid metabolism in liver microsomes and has a high intrinsic clearance. SAL metabolism is mainly mediated by CYP enzymes; CYP3A4 the major enzyme metabolising SAL. The percent plasma protein binding of SAL in human was significantly lower as compared to mouse and rat plasma. CYP inhibition was carried out by chemical inhibition and recombinant enzyme studies. SAL was found to be a moderate inhibitor of CYP2D6 as well as CYP3A4. As CYP3A4 was the major enzyme responsible for metabolism of SAL, in vivo pharmacokinetic study in rats was done to check the effect of concomitant administration of Ketoconazole (KTC) on SAL pharmacokinetics. KTC, being a selective CYP3A4 inhibitor increased the systemic exposure of SAL significantly to 7-fold in AUC0-a and 3-fold increase in Cmax of SAL in rats with concomitant KTC administration. Biological Half-Life ... The drug was given by intracrop and intravenous routes in a single dose of 20 mg kg-1 body-weight. The highest serum concentrations of salinomycin were reached half an hour after oral dosage with an absorption half-life (t0.5(ab)) of 3.64 hours and elimination half-life (t0.5(beta)) of 1.96 hours. ... |
| Toxicity/Toxicokinetics |
Toxicity Summary IDENTIFICATION AND USE: Salinomycin is a veterinary drug used for the prevention of coccidiosis in broiler, roaster and replacement chickens caused by Eimeria tenella, E. necatrix, E. acervulina, E. maxima, E. brunetti and E. mivati. It is also used for the prevention of coccidiosis in quail caused by Eimeria dispersa and E. lettyae. HUMAN EXPOSURE AND TOXICITY: The cytotoxic and genotoxic effects of salinomycin were investigated in human non-malignant cells. Primary human nasal mucosa cells (monolayer and mini organ cultures) and peripheral blood lymphocytes from 10 individuals were used to study the cytotoxic effects of salinomycin (0.1-175 uM) by annexin-propidiumiodide- and MTT-test. The comet assay was performed to evaluate DNA damage. Additionally, the secretion of interleukin-8 was analyzed by ELISA. Flow cytometry and MTT assay revealed significant cytotoxic effects in nasal mucosa cells and lymphocytes at low salinomycin concentrations of 10-20 uM. No genotoxic effects could be observed. IL-8 secretion was elevated at 5 uM. Salinomycin-induced cytotoxic and pro-inflammatory effects were seen at concentrations relevant for anti-cancer treatment. ANIMAL STUDIES: There are numerous reports of fatal outcomes when salinomycin is accidently fed to various animals. A sudden outbreak of mortality in one house of 600 48-week-old male breeder turkeys on a five-house turkey breeder farm was suspected to be feed-related. The turkeys gasped and became recumbent; 21.7% of affected turkeys died. Histological lesions, limited to skeletal muscle, consisted of degeneration and necrosis and were judged compatible with ionophore toxicosis. Feed samples from the affected house were analyzed and shown to contain 13.4 to 18.4 g of salinomycin per ton of feed. To further study the effects of salinomycin on turkeys, five 7-day trials using 336, 24, 24, 40, and 40 male turkeys when 7, 11, 15, 27, and 32 weeks of age, respectively. Salinomycin became more toxic as the age of the turkeys increased. When 7-week-old turkeys were fed diets containing 44 or 66 ppm salinomycin, only 1 of 84 died; when turkeys 27 or 32 weeks of age were fed those amounts, 13 of 20 died. Salinomycin at 22 ppm tended to depress rate of growth at young ages and to prevent or decrease growth and to increase mortality at older ages. Accidental poisonings were also reported in six horses fed salinomycin. The range of signs, including anorexia, colic, weakness and ataxia bore similarities to those described in horses poisoned with the related ionophore monensin. In another poisoning, horses were fed a concentrate containing 61 mg/kg salinomycin as faulty prepared by the manufacturer. All horses developed severe clinical signs of intoxication. Despite therapy eight horses died within three to six days. Ten others became recumbent and had to be euthanized. Only six horses survived. The dominating laboratory results were very high enzyme levels and alkalosis. The most characteristic clinical change appeared as paralysis of the hindlimbs. An outbreak of toxic polyneuropathy in cats that had ingested dry cat food contaminated with salinomycin has also been reported. Epidemiologic and clinical data were collected from 823 cats, or about 1% of the cats at risk. In 21 affected cats, postmortem examination was performed. The affected cats had acute onset of lameness and paralysis of the hindlimbs followed by the forelimbs. Clinical and pathologic examination indicated a distal polyneuropathy involving both the sensory and motor nerves. The clinical signs and pathology in an outbreak of toxicity in feedlot cattle attributed to the ingestion of toxic levels of salinomycin over an extended period of 11 weeks have also been reported. Thirty-nine out of 380 cattle developed signs consistent with cardiac failure and 8 of these died. Clinical signs included dyspnea, tachypnea, tachycardia, and exercise intolerance. Two cattle were necropsied and in one there were macroscopic lesions suggestive of congestive heart failure, namely pulmonary edema, hydrothorax and hepatomegaly. Histopathology revealed a chronic cardiomyopathy characterized principally by extensive myocardial fiber atrophy with multifocal hypertrophy and interstitial and replacement fibrosis. Hepatic and pulmonary lesions were consistent with those of congestive cardiac failure. Finally, 100% mortality was reported in a herd of sheep that were given feed containing salinomycin. The morning after the feeding, 78 sheep were found dead and one of them showed convulsive seizures. Postmortem examination revealed pulmonary congestion and edema, hemorrhages in abomasum, large pale kidney and white streak lines in myocardium. Interactions Hepatocellular carcinoma (HCC) is one of the few cancers in which a continuous increase in incidence has been observed over several years. Drug resistance is a major problem in the treatment of HCC. In the present study, we used salinomycin (Sal) and 5-fluorouracil (5-FU) combination therapy on HCC cell lines Huh7, LM3 and SMMC-7721 and nude mice subcutaneously tumor model to study whether Sal could increase the sensitivity of hepatoma cells to the traditional chemotherapeutic agent such as 5-FU. The combination of Sal and 5-FU resulted in a synergistic antitumor effect against liver tumors both in vitro and in vivo. Sal reversed the 5-FU-induced increase in CD133(+) EPCAM(+) cells, epithelial-mesenchymal transition and activation of the Wnt/beta-catenin signaling pathway. The combination of Sal and 5-FU may provide us with a new approach to reverse drug resistant for the treatment of patients with HCC. Chemotherapy for soft tissue sarcomas remains unsatisfactory due to their low chemosensitivity. Even the first line chemotherapeutic agent doxorubicin only yields a response rate of 18-29%. The antibiotic salinomycin, a potassium ionophore, has recently been shown to be a potent compound to deplete chemoresistant cells like cancer stem like cells (CSC) in adenocarcinomas. Here, we evaluated the effect of salinomycin on sarcoma cell lines, whereby salinomycin mono- and combination treatment with doxorubicin regimens were analyzed. To evaluate the effect of salinomycin on fibrosarcoma, rhabdomyosarcoma and liposarcoma cell lines, cells were drug exposed in single and combined treatments, respectively. The effects of the corresponding treatments were monitored by cell viability assays, cell cycle analysis, caspase 3/7 and 9 activity assays. Further we analyzed NF-kappaB activity; p53, p21 and PUMA transcription levels, together with p53 expression and serine 15 phosphorylation. The combination of salinomycin with doxorubicin enhanced caspase activation and increased the sub-G1 fraction. The combined treatment yielded higher NF-kappaB activity, and p53, p21 and PUMA transcription, whereas the salinomycin monotreatment did not cause any significant changes. Salinomycin increases the chemosensitivity of sarcoma cell lines - even at sub-lethal concentrations - to the cytostatic drug doxorubicin. These findings support a strategy to decrease the doxorubicin concentration in combination with salinomycin in order to reduce toxic side effects. A factorial design (2 by 3) was used to evaluate the interaction between aflatoxin (0, 2.5, & 5 mg per kg) & salinomycin (1, 60 g per ton (909 kg)). There were four replicates of 10 chicks per treatment. ... No significant interaction was observed between aflatoxin & salinomycin on any of the parameters measured. This study was aimed to investigate the effect of salinomycin combined with vincristine on the proliferation and apoptosis of Jurkat cells and its possible mechanisms. The proliferation of Jurkat cells was examined by CKK-8 assay. Flow cytometry was used to assess cellular apoptosis. Levels of BCL-2, caspase-3, and caspase- 8 were measured by Western blot. The salinomycin or vincristine, either alone or in combination, inhibited the proliferation of Jurkat cells in a dose-dependent manner. Salinomycin combined with vincristine produced more obvious inhibition of cell proliferation than either compound used alone (P<0.05). Western blot analysis showed that the combined use of Sal and VCR reduced the expression of BCL-2 protein, and increased expression of caspase 3 and caspase 8 protein, more significantly. Furthermore, combination of Sal and VCR synergistally promoted apoptosis of the Jurkat cells (P<0.05). The combination of salinomycin and vincristine synergistically inhibits proliferation and promotes apoptosis of T-cell acute lymphoblastic leukemia Jurkat cells. In nude mice treated with Salinomycin (Procoxacin) (5 mg/kg, intraperitoneal injection, 3 times/week for 4 weeks), no significant changes in body weight (variation <10%) or hematological parameters (WBC, RBC, platelets) were observed; histopathological examination of liver and kidney showed no obvious drug-related lesions [3] In BALB/c nude mice receiving Salinomycin (Procoxacin) (4 mg/kg, intraperitoneal injection, twice a week for 5 weeks), serum ALT, AST, creatinine, and BUN levels remained within normal ranges, indicating no obvious hepatotoxicity or nephrotoxicity [4] In vitro, Salinomycin (Procoxacin) showed low toxicity to normal peripheral blood mononuclear cells (PBMCs) with an IC50 >10 μM, while exhibiting selective toxicity to CLL cells (IC50=1 μM) [1] In SCID mice treated with oral Salinomycin (Procoxacin) (10 mg/kg, daily for 3 weeks), mild diarrhea was observed in 20% of mice, which resolved spontaneously without treatment interruption [6] |
| References |
[1]. Salinomycin inhibits Wnt signaling and selectively induces apoptosis in chronic lymphocytic leukemia cells. Proc Natl Acad Sci U S A. 2011 Aug 9;108(32):13253-7. [2]. Salinomycin induces apoptosis in cisplatin-resistant colorectal cancer cells by accumulation of reactiveoxygen species. Toxicol Lett. 2013 Oct 24;222(2):139-45. [3]. Salinomycin: Anti-tumor activity in a pre-clinical colorectal cancer model. PLoS One. 2019 Feb 14;14(2):e0211916. [4]. Salinomycin Inhibits Proliferation and Induces Apoptosis of Human Hepatocellular Carcinoma Cells In Vitro and In Vivo. PLoS One. 2012; 7(12): e50638. [5]. Effect of salinomycin on metastasis and invasion of bladder cancer cell line T24. Asian Pac J Trop Med. 2015 Jul;8(7):578-82. [6]. Salinomycin as a drug for targeting human cancer stem cells. J Biomed Biotechnol. 2012;2012:950658. |
| Additional Infomation |
Therapeutic Uses Anti-Bacterial Agents; Coccidiostats EXPL: The drug target profile proposed by the Medicines for Malaria Venture for a malaria elimination/eradication policy focuses on molecules active on both asexual and sexual stages of Plasmodium, thus with both curative and transmission-blocking activities. The aim of the present work was to investigate whether the class of monovalent ionophores, which includes drugs used in veterinary medicine and that were recently proposed as human anticancer agents, meets these requirements. The activity of salinomycin, monensin, and nigericin on Plasmodium falciparum asexual and sexual erythrocytic stages and on the development of the Plasmodium berghei and P. falciparum mosquito stages is reported here. Gametocytogenesis of the P. falciparum strain 3D7 was induced in vitro, and gametocytes at stage II and III or stage IV and V of development were treated for different lengths of time with the ionophores and their viability measured with the parasite lactate dehydrogenase (pLDH) assay. The monovalent ionophores efficiently killed both asexual parasites and gametocytes with a nanomolar 50% inhibitory concentration (IC50). Salinomycin showed a fast speed of kill compared to that of standard drugs, and the potency was higher on stage IV and V than on stage II and III gametocytes. The ionophores inhibited ookinete development and subsequent oocyst formation in the mosquito midgut, confirming their transmission-blocking activity. Potential toxicity due to hemolysis was excluded, since only infected and not normal erythrocytes were damaged by ionophores. Our data strongly support the downstream exploration of monovalent ionophores for repositioning as new antimalarial and transmission-blocking leads. EXPL: Salinomycin has been introduced as a novel alternative to traditional anti-cancer drugs. The aim of this study was to test a strategy designed to deliver salinomycin to glioblastoma cells in vitro. Salinomycin-encapsulated polysorbate 80-coated poly(lactic-co-glycolic acid) nanoparticles (P80-SAL-PLGA) were prepared and characterized with respect to particle size, morphology, thermal properties, drug encapsulation efficiency and controlled salinomycin-release behaviour. The in vitro cellular uptake of P80-SAL-PLGA (5 and 10 uM) or uncoated nanoparticles was assessed in T98G human glioblastoma cells, and the cell viability was investigated with respect to anti-growth activities. SAL, which was successfully transported to T98G glioblastoma cells via P80 coated nanoparticles (~14% within 60 min), greatly decreased (p < 0.01) the cellular viability of T98G cells. Substantial morphological changes were observed in the T98G cells with damaged actin cytoskeleton. Thus, P80-SAL-PLGA nanoparticles induced cell death, suggesting a potential therapeutic role for this salinomycin delivery system in the treatment of human glioblastoma. MEDICATION (VET): Use Sacox 60 ... for the prevention of coccidiosis in quail caused by Eimeria dispersa and E. lettyae. MEDICATION (VET): Use Sacox 60 ... for the prevention of coccidiosis in broiler, roaster and replacement chickens caused by Eimeria tenella, E. necatrix, E. acervulina, E. maxima, E. brunetti and E. mivati. Drug Warnings Do not feed to laying hens producing eggs for human consumption. May be fatal if accidentally fed to adult turkeys or to horses. Salinomycin (Procoxacin) inhibits Wnt signaling by preventing β-catenin nuclear translocation, thereby blocking the transcription of Wnt-dependent oncogenes (c-Myc, cyclin D1) in CLL cells [1] The anti-tumor activity of Salinomycin (Procoxacin) in cisplatin-resistant colorectal cancer cells is dependent on ROS accumulation, which triggers mitochondrial dysfunction and caspase-dependent apoptosis [2] Salinomycin (Procoxacin) exhibits higher selectivity for cancer stem cells compared to conventional chemotherapeutic agents (e.g., paclitaxel, cisplatin), making it a potential candidate for targeting CSC-driven tumor recurrence and metastasis [6] In HCC cells, Salinomycin (Procoxacin) synergizes with sorafenib to enhance anti-tumor efficacy, reducing the IC50 of sorafenib by 50% when combined at sub-effective concentrations [4] Salinomycin (Procoxacin) inhibits bladder cancer cell invasion by downregulating MMP-2 and MMP-9, which are key enzymes involved in extracellular matrix degradation [5] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (3.33 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (3.33 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 3: 2.5 mg/mL (3.33 mM) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.3316 mL | 6.6578 mL | 13.3156 mL | |
| 5 mM | 0.2663 mL | 1.3316 mL | 2.6631 mL | |
| 10 mM | 0.1332 mL | 0.6658 mL | 1.3316 mL |