carminomycin I (CA) is a selective inhibitor of CCRCC cell proliferation.
The similarity in the rate of change in XTT reduction of carminomycin I (CA) treated paired VHL−/− and VHL+/+ cells reveals that although carminomycin I (CA) impacts cell viability to a significant extent in VHL−/− cells, it did not impact the rate of change in viability to a similar extent compared to VHL+/+ cells. This suggests that the presence of pVHL impacts sensitivity to carminomycin I (CA) but not the rate of change in cell viability. We found that carminomycin I (CA) induced apoptosis in VHL−/− cells to a significant extent and that this activity was HIF and p53 independent. At low concentrations carminomycin I (CA) induced the activation of caspases 2 and 3 with little or no effect on the activation of caspases-8 or -9. This effect was accompanied by downregulation of the inhibitor of apoptosis protein survivin, a caspase-2 target. Although survivin levels appear to be steady at the 8 h time point while the level of caspase-2 is increasing in response to carminomycin I (CA) treatment, this does not necessarily imply that caspase-2 activation does not contribute to survivin down regulation. In fact, stress response to anthracyclines is known to result in the rapid induction of both mRNA and protein of inhibitor of apoptosis proteins including survivin which is followed by a gradual decrease of their expression (20). We also determined that carminomycin I (CA) was sequestered within the Golgi in CCRCC cells and that mediation by P-gp, which was also found within the Golgi, was required for its activity. However, whether carminomycin I (CA) is also sequestered within the Golgi of other P-gp expressing cell lines awaits further investigation. Its localization within the Golgi in CCRCC cells suggests that it might exert its action, at least in part, through effects on Golgi proteins. We, therefore, explored effects of carminomycin I (CA) on Golgi proteins and whether resistance to carminomycin I (CA) in VHL+/+ CCRCC cells was related to interaction of pVHL with Golgi proteins. We found that carminomycin I (CA) treatment resulted in the detection of the C-terminal fragment of p115 in the nucleus indicating that its apoptotic effects can in part be explained through its effects on p115 (21). Although carminomycin I (CA) had no effects on levels of the Golgi protein ERGIC-53, which is one of the few Golgi-proteins known to have a direct protein-protein interaction with pVHL, it impacted its distribution pattern between the ER and the Golgi. Our results also indicated that the secretion into the growth medium of A1AT, an ERGIC-53 client, was attenuated while intracellular A1AT levels increased in response to carminomycin I (CA) treatment. These are consistent with attenuation of function of ERGIC-53 (22).
carminomycin I (CA) is composed of an aglycone moiety and a monosaccharide residue. Since microsomes from mammalian tissues are known to metabolize anthracyclines to the superoxide anion of the aglycones and their monosaccharide residues through reductive glycosidic cleavage, it can be hypothesized that damage to Golgi proteins by the reactive oxygen species formed from the aglycone may be responsible for the effects of carminomycin I (CA) on proliferation in CCRCC cells (23-25). Alternatively, recognition of the monosaccharide residue in carminomycin I (CA) as a substrate may lead to attenuation of activity of Golgi proteins. Undoubtedly, however, carminomycin I (CA) also effects its activity through impact on additional pathways and targets. Demonstration of efficacy in xenograft models is pending preparative scale isolation of carminomycin I (CA) from the producing micro-organism.[1]

Following treatment with 0.1 μmol/L CA for 16 h, 0.5-1.0×106/ml live 786-O and 786-OVHL cells were harvested and stained with propidium iodide using Krishan’s buffer (0.1% sodium citrate, 0.02 mg/mL RNase A, 0.3% NP-40 and 50 μg/mL propidium iodide) at a pH of 7.4.[1]

[1]. Carminomycin I is an apoptosis inducer that targets the Golgi complex in clear cell renal carcinoma cells. Cancer Res. 2011 Jan 1;71(1):134-42.

Carminomycin is a toxic anthracycline antibiotic that is produced by Actinomadura carminata and also has potent antineoplastic activity. It has a role as an antineoplastic agent and an apoptosis inducer. It is an anthracycline antibiotic, an aminoglycoside antibiotic, a member of tetracenequinones, a member of p-quinones and a tertiary alpha-hydroxy ketone. It is a conjugate base of a carminomycin(1+). It derives from a hydride of a tetracene.
Carubicin is an anthracycline antineoplastic antibiotic isolated from the bacterium Actinomadura carminata. Carubicin intercalates into DNA and interacts with topoisomerase II, thereby inhibiting DNA replication and repair and RNA and protein synthesis.
A very toxic anthracycline-type antineoplastic related to DAUNORUBICIN, obtained from Actinomadura carminata.

C26H27NO10

513.49

513.163

C, 60.81; H, 5.30; N, 2.73; O, 31.16

50935-04-1

Carubicin hydrochloride;52794-97-5

443831

Red solid powder

1.6±0.1 g/cm3

750.6±60.0 °C at 760 mmHg

407.7±32.9 °C

0.0±2.6 mmHg at 25°C

1.727

3.54

6

11

3

37

944

6

O([C@]1([H])O[C@@H](C)[C@@H](O)[C@@H](N)C1)[C@H]1C[C@](O)(C(=O)C)CC2C(=C3C(C4=CC=CC(O)=C4C(=O)C3=C(O)C1=2)=O)O

XREUEWVEMYWFFA-CSKJXFQVSA-N

InChI=1S/C26H27NO10/c1-9-21(30)13(27)6-16(36-9)37-15-8-26(35,10(2)28)7-12-18(15)25(34)20-19(23(12)32)22(31)11-4-3-5-14(29)17(11)24(20)33/h3-5,9,13,15-16,21,29-30,32,34-35H,6-8,27H2,1-2H3/t9-,13-,15-,16-,21+,26-/m0/s1

(7S,9S)-9-acetyl-7-[(2R,4S,5S,6S)-4-amino-5-hydroxy-6-methyloxan-2-yl]oxy-4,6,9,11-tetrahydroxy-8,10-dihydro-7H-tetracene-5,12-dione

CMM; Antibiotic R 588A; Carubicin; carminomycin; Antibiotic R-588A; Antibiotic R 588-A; Carminomycin I; Demethyldaunomycin; Karminomitsin; karminomycin; Carminomicin I

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)