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ADDA 5 Hydrochloride

Alias: ADDA5 Hydrochloride ADDA 5 Hydrochloride ADDA-5 Hydrochloride
Cat No.:V5997 Purity: ≥98%
ADDA 5 HCl is a partially noncompetitive inhibitor of cytochrome c oxidase (CcO).
ADDA 5 Hydrochloride
ADDA 5 Hydrochloride Chemical Structure CAS No.: 473268-46-1
Product category: New1
This product is for research use only, not for human use. We do not sell to patients.
Size Price Stock Qty
5mg
10mg
Other Sizes
Official Supplier of:
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Product Description
ADDA 5 HCl is a partially noncompetitive inhibitor of cytochrome c oxidase (CcO). It can inhibit the activity of CcO isolated and purified from human glioma cells and bovine cardiomyocytes. The IC50s are respectively 18.93 μM and 31.82 μM.
Biological Activity I Assay Protocols (From Reference)
ln Vitro
Purified CcO from human glioblastoma and bovine heart had IC50 values of 18.93 μM and 31.82 μM for ADDA 5 HydroHClide, indicating that it is a somewhat non-competitive inhibitor of CcO. In GSCs generated from UTMZ and Jx22, ADDA 5 inhibits CcO activity with IC50 values of 21.4 ± 3.9 μM and 15.5 ± 2.8 μM, respectively. With an EC50 of 8.17 μM, ADDA 5 (25 μM) inhibits the development of UTMZ cells[1].
ln Vivo
At dosages up to 80 mg/kg, ADDA 5 does not induce observable toxicity in animals, and it effectively suppressed tumor growth in mice (8 mg/kg, ip) [1].
References
[1]. Oliva CR, et al. Identification of Small Molecule Inhibitors of Human Cytochrome c Oxidase That Target Chemoresistant Glioma Cells. J Biol Chem. 2016 Nov 11;291(46):24188-24199. Epub 2016 Sep 27
These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C24H36CLNO2
Molecular Weight
406.001146316528
Exact Mass
405.243
CAS #
473268-46-1
PubChem CID
2882960
Appearance
White to off-white solid powder
Hydrogen Bond Donor Count
2
Hydrogen Bond Acceptor Count
3
Rotatable Bond Count
7
Heavy Atom Count
28
Complexity
465
Defined Atom Stereocenter Count
0
SMILES
Cl.OC(CN1CCC2C(=CC=CC=2)C1)COCCC12CC3CC(C1)CC(C3)C2
InChi Key
YZSRZEWBOXEFRJ-UHFFFAOYSA-N
InChi Code
InChI=1S/C24H35NO2.ClH/c26-23(16-25-7-5-21-3-1-2-4-22(21)15-25)17-27-8-6-24-12-18-9-19(13-24)11-20(10-18)14-24;/h1-4,18-20,23,26H,5-17H2;1H
Chemical Name
1-[2-(1-adamantyl)ethoxy]-3-(3,4-dihydro-1H-isoquinolin-2-yl)propan-2-ol;hydrochloride
Synonyms
ADDA5 Hydrochloride ADDA 5 Hydrochloride ADDA-5 Hydrochloride
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

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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 : ~65 mg/mL (~160.10 mM)
H2O : ~2 mg/mL (~4.93 mM)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.12 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 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: ≥ 2.08 mg/mL (5.12 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.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (5.12 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.


 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 2.4631 mL 12.3153 mL 24.6305 mL
5 mM 0.4926 mL 2.4631 mL 4.9261 mL
10 mM 0.2463 mL 1.2315 mL 2.4631 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.

Calculator

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An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
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  • The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

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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  • The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box
g/mol

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Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
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Definitions of molecular mass, molecular weight, molar mass and molar weight:
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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
  • Counter screen for hits confirmation. A, representative visible absorption spectra of oxidized cyt c (0 μm ascorbate) and reduced cyt c after the addition of the indicated ascorbate concentrations. B, dose-response effect of ascorbate on cyt c reduction. Reduced cyt c concentration (μm) was calculated using absorbance values at 550 nm from the absorption spectra shown in A and the cyt c molar extinction coefficient of 1.92. Complete cyt c reduction was achieved by 40–50 μm ascorbate. C, the 20 ADDA compounds toxic to glioma cells (supplemental Table S1) were individually assayed to distinguish cyt c reducing compounds from true CcO inhibitors. The visible absorption spectra of cyt c after the addition of 50 μm concentrations of each compound was used to determine the percentage of cyt c reduction relative to cyt c reduction by 50 μm ascorbate (100%). Nine compounds (ADDA 2, 5, 8, 13, 15, 20, 21, 22, and 24) did not reduce cyt c and were, thus, selected for further analysis as true CcO inhibitors. Eleven compounds (ADDA 1, 3, 4, 6, 7, 10, 12, 14, 17, 19, and 23) did reduce cyt c and were identified as cyt c reductants and, therefore, not further investigated as CcO inhibitors.[1].Oliva CR, et al. Identification of Small Molecule Inhibitors of Human Cytochrome c Oxidase That Target Chemoresistant Glioma Cells. J Biol Chem. 2016 Nov 11;291(46):24188-24199. Epub 2016 Sep 27
  • Effects of ADDA 5 on mitochondrial complexes. A, ADDA 5 was tested on mitochondrial extracts from TMZ-resistant glioma cells for effects on the activity of complexes I-III, CcO (complex IV), and complex V of the mitochondrial transport chain. Scatter graphs represent the activity level of each complex in the presence of DMSO (Control) and ADDA 5 (25 μm). The results are averages from triplicate determinations from two independent experiments. ns, not significant; ***, p < 0.0002. B, representative SDS-PAGE stained with Coomassie Blue. Lane 1, protein ladder. Lane 2, purified CcO from bovine heart. Band identities were determined by MS. A band label of Mix indicates the presence of multiple CcO subunits due to incomplete denaturation of CcO. C, effect of ADDA 5 on the activity of purified CcO from bovine heart. ADDA 5 inhibited CcO with an IC50 of 31.82 ± 2.89 μm. D, representative SDS-PAGE stained with Coomassie Blue. Lane 1, protein ladder. Lane 2, purified CcO from human glioma cells. Band identities were determined by MS. A band label of Mix indicates the presence of multiple CcO subunits due to incomplete denaturation of CcO. E, effect of ADDA 5 on the activity of purified CcO from human glioma cells. ADDA 5 inhibited CcO with an IC50 of 18.93 ± 0.04 μm.[1].Oliva CR, et al. Identification of Small Molecule Inhibitors of Human Cytochrome c Oxidase That Target Chemoresistant Glioma Cells. J Biol Chem. 2016 Nov 11;291(46):24188-24199. Epub 2016 Sep 27
  • Inhibitory specificity of ADDA 5. The specificity of ADDA 5 was investigated by testing for inhibition of other enzymes, including XO (A), catalase (B), LDH (C), SOD (D), GPx (E), and GR (F). Activity assays were performed as described under “Experimental Procedures. No inhibition was observed at any tested concentration, suggesting that ADDA 5 is selective for CcO. Scatter plots are the results from duplicate determinations from two independent experiments.[1].Oliva CR, et al. Identification of Small Molecule Inhibitors of Human Cytochrome c Oxidase That Target Chemoresistant Glioma Cells. J Biol Chem. 2016 Nov 11;291(46):24188-24199. Epub 2016 Sep 27
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