13C labeled NAD+

Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as quantitative tracers while the drugs were being developed. Because deuteration may have an effect on a drug's pharmacokinetics and metabolic properties, it is a cause for concern [1].

[1]. Impact of Deuterium Substitution on the Pharmacokinetics of Pharmaceuticals. Ann Pharmacother. 2019 Feb;53(2):211-216.

[2]. Cellular and molecular mechanisms of metformin: an overview. Clin Sci (Lond), 2012. 122(6): p. 253-70.

[3]. Brandt, U., Energy converting NADH:quinone oxidoreductase (complex I). Annu Rev Biochem, 2006. 75: p. 69-92.

[4]. Kussmaul, L. and J. Hirst, The mechanism of superoxide production by NADH:ubiquinone oxidoreductase (complex I) from bovine heart mitochondria. Proc Natl Acad Sci U S A, 2006. 103(20): p. 7607-12.

NADH: Ubiquinone oxidoreductase (complex I) is a major source of reactive oxygen species in mitochondria and a significant factor in cellular oxidative stress. This paper describes the kinetics and molecular mechanism of superoxide anion generation from complex I isolated from bovine heart mitochondria, confirming that it primarily generates superoxide anions rather than hydrogen peroxide. Redox titration and electron paramagnetic resonance spectroscopy ruled out iron-sulfur clusters and flavin radicals as sources of superoxide anions, and superoxide anion generation was not enhanced during the turnover of complex I in the absence of a proton motive force. Therefore, superoxide anions are formed by the transfer of one electron from completely reduced flavin to O₂. The generated flavin radical is unstable, so the remaining electron may redistribute to the iron-sulfur cluster. The rate of superoxide anion generation depends on the bimolecular reaction between O₂ and reduced flavin in an empty active site. The proportion of flavin involved in the reaction is determined by a preequilibria, which is determined by the dissociation constants of NADH and NAD+ and the reduction potentials of flavin and NAD+. Therefore, the ratio and concentration of NADH and NAD+ determine the superoxide generation rate. This result clearly links the mechanism of our isolated enzyme to studies of intact mitochondria, in which superoxide production is enhanced when the NAD+ pool is reduced. Thus, our mechanism lays the foundation for constructing a causal relationship between complex I deficiency and pathological effects. [4] NADH:quinone oxidoreductase (complex I) pumps protons into the inner mitochondrial membrane or the plasma membrane of many bacteria. Human complex I is involved in a variety of pathological conditions and degenerative processes. Complex I consists of 14 central subunits and up to 32 accessory subunits and is one of the largest membrane-bound protein complexes. The peripheral arms of the L-shaped molecule contain flavin mononucleotides and eight or nine iron-sulfur clusters as redox cofactors. Seven of these iron-sulfur clusters form a linear electron transport chain between flavin and quinone. In most organisms, the seven most hydrophobic subunits that make up the core of the membrane arm are encoded by the mitochondrial genome. Most of the central subunits evolved from subunits of different hydrogenases and bacterial Na+/H+ antitransporters. This evolutionary origin is reflected in the three functional modules of complex I. The coupling mechanism of complex I likely involves semiquinone intermediates that drive proton pumping through redox-related conformational changes. [3]

C1613C5H27N7O14P2

668.388380289078

668.125

1859096-06-2

NAD+;53-84-9;NAD+-d4

168006970

White to off-white solid powder

-6

7

18

11

44

1120

8

C1=CC(=C[N+](=C1)[13C@H]2[13C@@H]([13C@@H]([13C@H](O2)[13CH2]OP(=O)([O-])OP(=O)(O)OC[C@@H]3[C@H]([C@H]([C@@H](O3)N4C=NC5=C(N=CN=C54)N)O)O)O)O)C(=O)N

BAWFJGJZGIEFAR-ILYRTYENSA-N

InChI=1S/C21H27N7O14P2/c22-17-12-19(25-7-24-17)28(8-26-12)21-16(32)14(30)11(41-21)6-39-44(36,37)42-43(34,35)38-5-10-13(29)15(31)20(40-10)27-3-1-2-9(4-27)18(23)33/h1-4,7-8,10-11,13-16,20-21,29-32H,5-6H2,(H5-,22,23,24,25,33,34,35,36,37)/t10-,11-,13-,14-,15-,16-,20-,21-/m1/s1/i5+1,10+1,13+1,15+1,20+1

[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2R,3S,4R,5R)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxy(2,3,4,5-13C4)oxolan-2-yl](113C)methyl phosphate

NAD+-13C5-1; 1859096-06-2; NICOTINAMIDE ADENINE DINUCLEOTIDE (NAD+), NH4 SALT (RIBOSE-13C5, 98%) CHEMICAL PURITY 95%; HY-B0445S1; CS-0565524; [[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2R,3S,4R,5R)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxy(2,3,4,5-13C4)oxolan-2-yl](113C)methyl phosphate; 1-((2R,3R,4S,5R)-5-((((((((2R,3S,4R,5R)-5-(6-Amino-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)oxy)oxidophosphoryl)oxy)methyl-13C)-3,4-dihydroxytetrahydrofuran-2-yl-2,3,4,5-13C4)-3-carbamoylpyridin-1-ium

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.)