Costunolide (NSC 106404)

Alias: Costus lactone; CCRIS 6754; NSC 106404; CCRIS-6754; NSC-106404; CCRIS6754; NSC106404; Costunolide; Costus lactone; (+)-Costunolide
Cat No.:V1388 Purity: ≥98%
Costunolide [also known as (+)-Costunolide, Costus lactone, CCRIS-6754 and NSC-106404] isa naturally occurring sesquiterpene compound with a wide spectrum of biological activities such as anti-inflammatory, anticancer, antidiabetic, antiviral, antimicrobial, antiulcer,antifungal, antioxidant, and anthelmintic properties.
Costunolide (NSC 106404) Chemical Structure CAS No.: 553-21-9
Product category: Telomerase
This product is for research use only, not for human use. We do not sell to patients.
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Purity & Quality Control Documentation

Purity: ≥98%

Product Description

Costunolide [also known as (+)-Costunolide, Costus lactone, CCRIS-6754 and NSC-106404] is a naturally occurring sesquiterpene compound with a wide spectrum of biological activities such as anti-inflammatory, anticancer, antidiabetic, antiviral, antimicrobial, antiulcer, antifungal, antioxidant, and anthelmintic properties. It was initially discovered in 1960 in Saussurea costus roots. With an IC50 of 20 μM, costunolide inhibits FPTase. It also inhibits telomerase, with an IC50 of 65-90 μM.

Biological Activity I Assay Protocols (From Reference)
Targets
FPTase ( IC50 = 20 μM ); Telomerase (MDA-MB-231) ( IC50 = 65 μM ); Telomerase (MCF-7) ( IC50 = 90 μM )
ln Vitro

In vitro activity: Costunolide has a concentration- and time-dependent effect on the growth and telomerase activity of MCF-7 and MDA-MB-231 cells. As mentioned earlier, costunolide also, in a dose-dependent manner, blocks farnesyl–protein transferase (FPTase) from farnesylating human lamin-B. Human tumor cell proliferation (A549, SK-OV-3, SK-MEL-2, XF498 and HCT-1) will be markedly suppressed in a dose-dependent manner by continuous treatment with Costunolide for 48 hours.[2] In HL-60 human leukemia cells, costunolide cause apoptosis via cytochrome C release into the cytosol and ROS-mediated mitochondrial permeability transition.[3] Including Trichophyton mentagrophytes, T.simlii, T.rubrum, and others, a recent study found that costunolide demonstrates strong antifungal activity.[4]

ln Vivo
Costunolide blocks the angiogenic factor signaling pathway, which in turn inhibits the angiogenic response. Costunolide inhibits VEGF-stimulated neovascularization in mice using a corneal micropocket assay. [5]
Enzyme Assay
The TRAP assay measures telomerase activity using the TRAPez Telomerase Detection Kit, which contains primers for a 36-bp internal control (IC) that allows the amplification of telomerase activity to be quantified within a linear range near 2.5 logs. To treat the extract with RNase, 10μL of it are incubated for 20 minutes at 37 °C with 1μg of RNase. The TRAP assay products are separated by electrophoresis in a nondenaturing 12% PAGE with a buffer that contains 0.5 × Tris-borate EDTA, and they are then autoradiographically detected. Dried gels are exposed to room temperature Fuji Imaging Plates in order to quantify TRAP products. The untreated control cell signal is assigned a standard value of 100 after background correction is applied to the results. The signal intensities of cells treated with costunolide are expressed as a percentage of the maximum value of 100 and are compared to the standard.[1]
Cell Assay
1) In a 96-well plate, plate 500–10,000 cells with 200 μL of media per well. 8 wells should be left empty for blank controls. 2) To enable the cells to adhere to the wells, incubate (37 °C, 5% CO2) for the entire night. 3) Fill each well with 2 μL of dissolved costunolide in DMSO. To ensure that the samples and media are well mixed, place on a shaking table and shake for five minutes at 150 rpm. 5) Allow Costunolide to work for 48 hours by incubating at 37 °C with 5% CO2. 6) Add at least 2 milliliters (mL) of MTT solution (5 mg/mL) to each 96-well plate in PBS. Since MTT in solution is not long-term stable, do not create a stock. 7) Fill each well with 20 μL of MTT solution. Put the media on a shaking table and spin it at 150 rpm for five minutes to fully incorporate the MTT. 8) Let the MTT metabolize for one to five hours by incubating at 37 °C with 5% CO2. 9) Dismiss the press. (If needed, dry the plate on paper towels to get rid of any residue. The MTT metabolic product, formazan, should be redissolved in 200 μL of DMSO. To completely combine the formazan and solvent, put on a shaking table and shake for five minutes at 150 rpm. 11) Determine the optical density at 560 nm. 11) Subtract background at 670 nm and read optical density at 560 nm. There should be a direct relationship between cell quantity and optical density.
Animal Protocol
Dissolved in DMSO; 100 mg/kg; i.p. injection
Hydron N containing VEGF are implanted into mouse cornea.
References

[1]. Cancer Lett . 2005 Sep 28;227(2):153-62.

[2]. Planta Med . 2001 Jun;67(4):358-9.

[3]. Biol Pharm Bull . 2001 Mar;24(3):303-6.

[4]. BMC Complement Altern Med . 2012 Mar 7:12:13.

[5]. Cancer Lett . 2002 Dec 10;187(1-2):129-33.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C15H20O2
Molecular Weight
232.32
Exact Mass
232.15
Elemental Analysis
C, 77.55; H, 8.68; O, 13.77
CAS #
553-21-9
Related CAS #
553-21-9
Appearance
Solid powder
SMILES
C/C/1=C\CC/C(=C/[C@@H]2[C@@H](CC1)C(=C)C(=O)O2)/C
InChi Key
HRYLQFBHBWLLLL-AHNJNIBGSA-N
InChi Code
InChI=1S/C15H20O2/c1-10-5-4-6-11(2)9-14-13(8-7-10)12(3)15(16)17-14/h5,9,13-14H,3-4,6-8H2,1-2H3/b10-5+,11-9+/t13-,14+/m0/s1
Chemical Name
(3aS,6E,10E,11aR)-6,10-dimethyl-3-methylidene-3a,4,5,8,9,11a-hexahydrocyclodeca[b]furan-2-one
Synonyms
Costus lactone; CCRIS 6754; NSC 106404; CCRIS-6754; NSC-106404; CCRIS6754; NSC106404; Costunolide; Costus lactone; (+)-Costunolide
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

Shipping Condition
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
Solubility Data
Solubility (In Vitro)
DMSO: ~47 mg/mL (~202.3 mM)
Water: <1 mg/mL
Ethanol: 1~25 mg/mL (4.3~107.6 mM)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.5 mg/mL (10.76 mM) (saturation unknown) in 10% EtOH + 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 EtOH 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 (10.76 mM) (saturation unknown) in 10% EtOH + 90% (20% SBE-β-CD in 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 EtOH stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (10.76 mM) (saturation unknown) in 10% EtOH + 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 EtOH stock solution to 900 μL of corn oil and mix evenly.


Solubility in Formulation 4: ≥ 2.08 mg/mL (8.95 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 20.8 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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 5: ≥ 2.08 mg/mL (8.95 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
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.

Solubility in Formulation 6: ≥ 2.08 mg/mL (8.95 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

Solubility in Formulation 7: 2% DMSO +Corn oil : 10 mg/mL

 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 4.3044 mL 21.5220 mL 43.0441 mL
5 mM 0.8609 mL 4.3044 mL 8.6088 mL
10 mM 0.4304 mL 2.1522 mL 4.3044 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.

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What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
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In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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Calculation results

Working concentration mg/mL;

Method for preparing DMSO stock solution mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.

Method for preparing in vivo formulation:Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.

(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
             (2) Be sure to add the solvent(s) in order.

Biological Data
  • Costunolide (1) and Eremanthin (2). BMC Complement Altern Med . 2012 Mar 7:12:13.
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