molecular formula C6H6N2O B132304 Nicotinamide-d4 CAS No. 347841-88-7

Nicotinamide-d4

Cat. No.: B132304
CAS No.: 347841-88-7
M. Wt: 126.15 g/mol
InChI Key: DFPAKSUCGFBDDF-RHQRLBAQSA-N
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Scientific Research Applications

Nicotinamide-d4 has a wide range of applications in scientific research:

    Chemistry: Used as a tracer in metabolic studies to understand the pathways and mechanisms of nicotinamide metabolism.

    Biology: Employed in studies of cellular redox states and the role of NAD+ in cellular processes.

    Medicine: Investigated for its potential therapeutic effects in conditions related to NAD+ deficiency, such as aging and neurodegenerative diseases.

    Industry: Utilized in the development of deuterated drugs with improved pharmacokinetic properties.

Safety and Hazards

Nicotinamide-d4 is generally safe, but potential risks for epigenetic alterations are associated with chronic use at high doses . Minor abnormalities of liver enzymes can infrequently occur at the doses used for diabetes prevention .

Future Directions

Future pre-clinical and clinical studies are needed to evaluate and validate the therapeutic potential of Nicotinamide-d4 and other NAD+ precursors . New studies will benefit from comparing the effectiveness of different NAD+ precursors side by side .

Biochemical Analysis

Biochemical Properties

Nicotinamide-d4 plays a pivotal role in NAD+ synthesis, contributing to redox reactions and energy production in cells . It interacts with various enzymes, proteins, and other biomolecules. For instance, it is a precursor in the synthesis of the metabolic cofactor NAD+ and an inhibitor of sirtuin 1 (SIRT1) . It increases the activity of serine palmitoyltransferase (SPT) and the biosynthesis of ceramide, glucosylceramide, sphingomyelin, free fatty acids, and cholesterol in primary human keratinocytes .

Cellular Effects

This compound has significant effects on various types of cells and cellular processes. At a concentration of 10 µM, it increases the activity of serine palmitoyltransferase (SPT) and the biosynthesis of ceramide, glucosylceramide, sphingomyelin, free fatty acids, and cholesterol in primary human keratinocytes . At a concentration of 40 µM, it induces apoptosis in SNU-398, SNU-739, and HepG2 hepatocellular carcinoma (HCC) cells .

Molecular Mechanism

This compound exerts its effects at the molecular level through various mechanisms. It serves as a precursor in the synthesis of the metabolic cofactor NAD+ and inhibits sirtuin 1 (SIRT1), a class of enzymes that catalyze non-redox reactions . It also increases the activity of serine palmitoyltransferase (SPT), leading to the biosynthesis of various lipids .

Temporal Effects in Laboratory Settings

It is known that NAD+ and its precursors, such as this compound, are unstable in blood and difficult to measure

Metabolic Pathways

This compound is involved in the NAD+ synthesis pathway . It is a precursor in the synthesis of the metabolic cofactor NAD+

Preparation Methods

Synthetic Routes and Reaction Conditions: The synthesis of nicotinamide-d4 typically involves the deuteration of nicotinamide. One common method is the catalytic exchange of hydrogen atoms with deuterium using deuterium gas (D2) in the presence of a suitable catalyst such as palladium on carbon (Pd/C). The reaction is carried out under controlled temperature and pressure conditions to ensure complete deuteration.

Industrial Production Methods: Industrial production of this compound follows similar synthetic routes but on a larger scale. The process involves the use of high-pressure reactors and continuous flow systems to achieve efficient deuteration. The purity of the final product is ensured through rigorous quality control measures, including high-performance liquid chromatography (HPLC) and mass spectrometry.

Chemical Reactions Analysis

Types of Reactions: Nicotinamide-d4 undergoes various chemical reactions, including:

    Oxidation: this compound can be oxidized to nicotinic acid-d4 using oxidizing agents such as potassium permanganate (KMnO4) or chromium trioxide (CrO3).

    Reduction: It can be reduced to 1,4-dihydrothis compound using reducing agents like sodium borohydride (NaBH4).

    Substitution: Halogenation reactions can introduce halogen atoms into the this compound molecule using reagents like bromine (Br2) or chlorine (Cl2).

Common Reagents and Conditions:

    Oxidation: Potassium permanganate in acidic or neutral conditions.

    Reduction: Sodium borohydride in methanol or ethanol.

    Substitution: Halogenation using bromine or chlorine in the presence of a catalyst.

Major Products Formed:

    Oxidation: Nicotinic acid-d4.

    Reduction: 1,4-Dihydrothis compound.

    Substitution: Halogenated derivatives of this compound.

Comparison with Similar Compounds

    Nicotinamide: The non-deuterated form of nicotinamide.

    Nicotinamide mononucleotide (NMN): A direct precursor to NAD+.

    Nicotinamide riboside (NR): Another NAD+ precursor with similar applications.

Uniqueness: Nicotinamide-d4 is unique due to its deuterium labeling, which provides enhanced stability and allows for detailed metabolic studies. The deuterium atoms also result in a slightly altered pharmacokinetic profile, which can be advantageous in certain therapeutic applications .

Properties

IUPAC Name

2,4,5,6-tetradeuteriopyridine-3-carboxamide
Source PubChem
URL https://pubchem.ncbi.nlm.nih.gov
Description Data deposited in or computed by PubChem

InChI

InChI=1S/C6H6N2O/c7-6(9)5-2-1-3-8-4-5/h1-4H,(H2,7,9)/i1D,2D,3D,4D
Source PubChem
URL https://pubchem.ncbi.nlm.nih.gov
Description Data deposited in or computed by PubChem

InChI Key

DFPAKSUCGFBDDF-RHQRLBAQSA-N
Source PubChem
URL https://pubchem.ncbi.nlm.nih.gov
Description Data deposited in or computed by PubChem

Canonical SMILES

C1=CC(=CN=C1)C(=O)N
Source PubChem
URL https://pubchem.ncbi.nlm.nih.gov
Description Data deposited in or computed by PubChem

Isomeric SMILES

[2H]C1=C(C(=C(N=C1[2H])[2H])C(=O)N)[2H]
Source PubChem
URL https://pubchem.ncbi.nlm.nih.gov
Description Data deposited in or computed by PubChem

Molecular Formula

C6H6N2O
Source PubChem
URL https://pubchem.ncbi.nlm.nih.gov
Description Data deposited in or computed by PubChem

DSSTOX Substance ID

DTXSID20574036
Record name (~2~H_4_)Pyridine-3-carboxamide
Source EPA DSSTox
URL https://comptox.epa.gov/dashboard/DTXSID20574036
Description DSSTox provides a high quality public chemistry resource for supporting improved predictive toxicology.

Molecular Weight

126.15 g/mol
Source PubChem
URL https://pubchem.ncbi.nlm.nih.gov
Description Data deposited in or computed by PubChem

CAS No.

347841-88-7
Record name (~2~H_4_)Pyridine-3-carboxamide
Source EPA DSSTox
URL https://comptox.epa.gov/dashboard/DTXSID20574036
Description DSSTox provides a high quality public chemistry resource for supporting improved predictive toxicology.
Record name 347841-88-7
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Synthesis routes and methods I

Procedure details

The reaction was carried out in a reaction mixture (30 ml), comprising 500 mM 3-cyanopyridine, 40 mM potassium phosphate buffer (pH 7.0) and resting cells (dry weight 2.3 mg). During the reaction, 3-cyanopyridine (500 mM) was added 7 times as soon as it was consumed. In this manner, 4.0 M 3-cyanopyridine was added in the course of 15 h and 3.89 M (475 g/l) nicotinamide was formed, corresponding to a yield of 97.3%. Nicotinic acid was not formed.
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97.3%

Synthesis routes and methods II

Procedure details

High performance liquid chromatography was used to detect Nampt reaction products. HPLC was performed with Waters 515 pumps and a 2487 detector (Waters, Mass.) with a Supelco LC-18-T column (15 cm×4.6 cm; Supelco, Pa.). The Nampt reaction was conducted at 37° C. for 15 min in 500 μl of reaction buffer (50 mM Tris-HCl [pH 7.5], 10 mM MgCl2, 50 mM nicotinamide, 0.2 mM PRPP) with 50 μg of the recombinant Nampt protein. The reaction was terminated by adding 125 μl of 1 M HClO4. Protein was then precipitated at 18,000 g, and 500 μl of the supernatant was neutralized with 40 μl of 3 M K2CO3. After centrifugation, 100 μl of sample was mixed with 400 μl of Buffer A (50 mM K2PO4/KHPO4, pH 7.0) and loaded into the HPLC system. The products from Nampt reaction were monitored by absorbance at 261 nm. Results of HPLC detection of Nampt reaction products showed that the mouse Nampt produced nicotinamide mononucleotide (NMN) from nicotinamide and PRPP (see, e.g., FIG. 4D). Nampt failed to catalyze the synthesis of nicotinic acid mononucleotide (NaMN) from nicotinic acid and PRPP (see, e.g., FIGS. 13A and 13B), confirming the substrate specificity of this enzyme. In isolated reactions, it was also confirmed that Nmnat catalyzed the synthesis of NAD from NMN and ATP.
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Synthesis routes and methods III

Procedure details

0.096 mole of 3-cyanopyridine was dissolved in 5.556 mole of water and 0.0115 mole of MnO2, prepared by above method, was added to this. The mixture was refluxed at 105° C. for 8 hrs. The reaction mixture was cooled and filtered. The filtrate was evaporated in dryness to get solid nicotinamide 0.095 mole. Yield of isonicotinamide was 98.9 mole %.
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Synthesis routes and methods IV

Procedure details

In this Example a USP grade nicotinamide product was recovered from a crude nicotinamide product medium. The crude contained 39.7% nicotinamide, 2.15% sodium nicotinate and 0.14% 3-cyanopyridine. These and all other percentages given in this Example are percentages by weight unless indicated otherwise. The total sodium content of the medium was 0.38%. On a dry basis, the reaction crude contained 93.7% nicotinamide, 5.07% sodium nicotinate, 0.33% 3-cyanopyridine, and 0.9% total sodium. Cation and weak base resins were utilized in the recovery process. The cation exchange resin was Dowex Marathon C, a sulfonated copolymer of styrene and divinylbenzene (gel form). The weak base resin was Dowex Marathon WBA, a dimethylamine-functionalized chloromethylated copolymer of styrene and divinylbenzene (macroporous form with a monodisperse size distribution). After washing with deionized water, these resins were loaded into columns each having an inner diameter of 15 mm and a height of 30 cm, leaving about 1.5 inches head space at the top of the columns. The reaction crude was then successively treated over the cation-exchange resin (at 28 ml/min), the weak base resin (20 ml/min), the cation-exchange resin (28 ml/min), and the weak base resin (20 ml/min). The cation-exchange resin was regenerated after every ten bed volumes of reaction crude, by a cycle that included a water wash (20 ml/min, 1.25 bed volumes), a 12% sulfuric acid strip (7 ml/min, 1 bed volume), and another water wash (20 ml/min, 1.25 bed volumes). The weak base resin was regenerated after every five bed volumes of reaction crude, by a cycle including a water wash (20 ml/min, 1.6 bed volumes), a 4% sodium hydroxide strip (20 ml/min, 1 bed volume), and another water wash (2.6 bed volumes). The feeds were analyzed by HPLC after the first pass cation-exchange and weak base, and second pass cation-exchange and weak base. Typical results from such experiments are presented in Table 1 below, top. The lower section of Table 1 gives a typical product analysis on a water free basis. The extreme right hand column of Table 1 sets out the results of an analysis of the product after recovery by evaporation (no crystallization performed). As can be seen, this processing reduced the 0.9% initial sodium to an undetectable level, and the initial 5.07% nicotinate to 0.13% (as nicotinic acid) on a dry weight basis, providing a USP grade nicotinamide.
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Retrosynthesis Analysis

AI-Powered Synthesis Planning: Our tool employs the Template_relevance Pistachio, Template_relevance Bkms_metabolic, Template_relevance Pistachio_ringbreaker, Template_relevance Reaxys, Template_relevance Reaxys_biocatalysis model, leveraging a vast database of chemical reactions to predict feasible synthetic routes.

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Strategy Settings

Precursor scoring Relevance Heuristic
Min. plausibility 0.01
Model Template_relevance
Template Set Pistachio/Bkms_metabolic/Pistachio_ringbreaker/Reaxys/Reaxys_biocatalysis
Top-N result to add to graph 6

Feasible Synthetic Routes

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Customer
Q & A

Q1: What is the role of Nicotinamide-d4 in the analysis of water-soluble vitamins?

A1: In the study "Fast simultaneous determination of multiple water-soluble vitamins and vitamin-like compounds in infant formula by UPLC-MS/MS" [], this compound serves as an internal standard. Internal standards are compounds with similar chemical properties to the analytes of interest but are not present in the original sample.

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