Peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) – upregulation of expression; β-secretase 1 (BACE1) – promotion of ubiquitination and proteasomal degradation; Nicotinamide adenine dinucleotide (NAD+) – precursor leading to increased steady-state levels. [2]

The acetylation status of SOD2 and Ndufa9 is decreased by nicotinamide riboside (0.5 nM; 24 hours) [1]. In C2C12, Hepa1.6, and HEK293 cells, nicotinamide riboside increases intracellular and mitochondrial NAD+ content in a concentration-dependent manner within the concentration range of 1-1000 μM [1]. Nicotinamide riboside supports innate immunity against coronavirus (CoV), one of the causes of COVID-19, and improves NAD and antiviral poly(ADP-ribose) polymerase (PARP) function [3].

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In primary cortical-hippocampal neuron cultures derived from Tg2576 mouse embryos, treatment with Nicotinamide riboside significantly increased PGC-1α protein levels, as determined by Western blot analysis. This increase was abolished by infection with adenoviral PGC-1α shRNA for gene silencing. [2]
In the same primary neuronal cultures, Nicotinamide riboside treatment decreased BACE1 protein levels. This decrease was largely abolished by PGC-1α silencing via adenoviral PGC-1α shRNA, indicating that the effect on BACE1 is mediated through PGC-1α. [2]
In HEK293 cells stably transfected with Myc-tagged BACE1 (Myc-BACE1), treatment with Nicotinamide riboside or overexpression of PGC-1α via adenoviral infection increased the levels of ubiquitinated BACE1 protein, as detected by immunoprecipitation followed by Western blot with an anti-ubiquitin antibody. This suggests NR promotes BACE1 ubiquitination. Concurrently, levels of monoubiquitin were decreased in cells treated with NR or overexpressing PGC-1α. [2]

Nicotinamide riboside (oral; 400 mg/kg/day; for 16 weeks) raises intracellular and plasma NAD+ content in a tissue-specific way when taken long-term [1].

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In 7- to 8-month-old Tg2576 Alzheimer's disease model mice, dietary treatment with 250 mg/kg/day of Nicotinamide riboside in drinking water for 3 months significantly improved cognitive performance in the novel object recognition test. Treated mice spent 63.2 ± 1.7% of time exploring the novel object compared to 42.0 ± 9.2% in non-treated controls. [2]
The same treatment regimen (250 mg/kg/day NR for 3 months) significantly increased the steady-state levels of NAD+ in the cerebral cortex of Tg2576 mice. [2]
Quantitative RT-PCR analysis showed that this NR treatment significantly increased PGC-1α mRNA levels in the brain of Tg2576 mice compared to untreated controls. [2]
Enzyme-linked immunosorbent assay (ELISA) showed that levels of Aβ1-42 were significantly decreased in the brains of Tg2576 mice treated with NR compared to placebo-treated controls. [2]
In hippocampal slices from Tg2576 mice, application of 20 μM Nicotinamide riboside for 4 hours abolished the deficits in long-term potentiation (LTP) recorded in the CA1 region. The LTP response was 224 ± 15% of baseline at 120 minutes post-tetanus in NR-treated slices versus 164 ± 12% in control slices. NR perfusion did not affect basal synaptic transmission or LTP in slices from wild-type mice. [2]
Quantitative RT-PCR analysis on cerebral cortex extracts from Tg2576 mice treated with NR for 3 months showed significant upregulation of mRNA levels for several mitochondrial energy metabolism-related genes, including citrate synthase, aconitase, pyruvate dehydrogenase kinase 3, cytochrome c subunit VIc, phosphoglycerate kinase 1, and glucose phosphate isomerase 1. [2]

Western Blot Analysis[1]
Cell Types: HEK293T Cell
Tested Concentrations: 0.5 nM
Incubation Duration: 24 hrs (hours)
Experimental Results: diminished acetylation status of Ndufa9 and SOD2.

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Primary cortical-hippocampal neuronal cultures were prepared from the brains of 14.5-day-old embryos from Tg2576 mice. Dissociated cells were seeded and maintained in neural basal medium supplemented with specific growth factors. After 7 days in vitro, cells were used for treatment. [2]
For Western blot analysis, cells or brain tissues were lysed in appropriate lysis buffers supplemented with protease inhibitors. Protein lysates were separated by SDS-PAGE, transferred to nitrocellulose membranes, and probed with primary antibodies against target proteins (e.g., PGC-1α, BACE1, ubiquitin), followed by HRP-conjugated secondary antibodies and detection by enhanced chemiluminescence. [2]
For quantitative RT-PCR, total RNA was extracted from tissues or cells using a commercial kit. Complementary DNA was synthesized using a reverse transcription supermix. Quantitative PCR was performed using SYBR Green master mix on a real-time PCR system to measure mRNA levels of target genes. [2]
For the BACE1 ubiquitination assay, HEK293 cells stably expressing Myc-BACE1 were treated with NR and/or infected with adenoviral vectors. Cell lysates were immunoprecipitated using an anti-BACE1 antibody, and the precipitated proteins were analyzed by Western blot using an anti-ubiquitin antibody to detect ubiquitinated BACE1. [2]

Animal/Disease Models: 10weeks old C57Bl/6J mice [1]
Doses: 400 mg/kg
Route of Administration: oral; daily; continued for 16 weeks
Experimental Results: Plasma and intracellular NAD+ levels increased in a tissue-specific manner.

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Seven- to 8-month-old Tg2576 transgenic mice were treated with Nicotinamide riboside at a dose of 250 mg/kg/day, administered via drinking water. The treatment started when mice were 5-6 months old and continued for 3 months until they were 10-11 months old. Control Tg2576 mice received saline in their drinking water. [2]
After the treatment period, cognitive function was assessed using the novel object recognition test. Mice were first habituated to an apparatus and allowed to explore a familiar object. After a delay, they were reintroduced to the apparatus containing the familiar object and a novel object. The time spent exploring each object was recorded. [2]
After behavioral testing, mice were sacrificed. One brain hemisphere was snap-frozen for biochemical analyses (e.g., NAD+ assay, ELISA, Western blot, qRT-PCR), and the other hemisphere was fixed for histopathological examination. [2]
For electrophysiology studies, hippocampal slices (400 μm thick) were prepared from Tg2576 or wild-type mice. Slices were maintained in an interface chamber with artificial cerebrospinal fluid. After recovery, slices were perfused with 20 μM Nicotinamide riboside for 4 hours. Basal synaptic transmission and long-term potentiation were recorded in the CA1 region following standard protocols. [2]

This study showed that daily intake of 250 mg/kg nicotinamide nucleoside (NRI) for 3 months significantly increased NAD+ homeostasis in the cerebral cortex of Tg2576 mice. [2]

[1]. The NAD(+) precursor nicotinamide riboside enhances oxidative metabolism and protectsagainst high-fat diet-induced obesity. Cell Metab. 2012 Jun 6;15(6):838-47.

[2]. Nicotinamide Riboside Restores Cognition Through an Upregulation of Proliferator-Activated Receptor-γ Coactivator 1α Regulated β-Secretase 1 Degradation and Mitochondrial Gene Expression in Alzheimer's Mouse Models. Neurobiol Aging. 2013 Jun;34(6):1581-8.

[3]. Coronavirus and PARP Expression Dysregulate the NAD Metabolome: A Potentially Actionable Component of Innate Immunity. bioRxiv. 2020 Apr 30;2020.04.17.047480.

N-ribosylnicotinamide is a pyridine nucleoside composed of nicotinamide and a β-D-furanose moiety at the 1-position. It is a human metabolite, a Saccharomyces cerevisiae metabolite, a mouse metabolite, and an anti-aging agent. It is an N-glycosylnicotinamide and pyridine nucleoside. Nicotinamide nucleoside is being studied in the clinical trial NCT03432871 (Nicotinamide Nucleoside and Mitochondrial Biosynthesis). Nicotinamide nucleoside is a metabolite found or produced in Escherichia coli (K12 strain, MG1655 strain). Nicotinamide nucleoside has been reported in fruit flies, humans, and other organisms with relevant data. Nicotinamide nucleoside is an orally effective form of vitamin B3 and a precursor to nicotinamide adenine dinucleotide (NAD+), with potential use in treating chemotherapy-induced peripheral neuropathy (CIPN). Following oral administration, nicotinamide nucleoside (NR) is converted to nicotinamide mononucleotide (NMON) by NR kinases—nicotinamide nucleoside kinase 1 (NRK 1) and nicotinamide nucleoside kinase 2 (NRK 2). NMON is then converted to a second adenine via nicotinamide mononucleotide adenylate transferase, generating NAD+. NAD+ is an important redox coenzyme that may protect against axonal damage caused by mechanical and neurotoxic injuries. Maintaining NAD+ levels may also protect against mitochondrial diseases. NR may help increase and maintain NAD+ levels, thereby improving potential mechanisms associated with the development of chemotherapy-induced peripheral neuropathy (CIPN), including mitochondrial dysfunction and peripheral neurodegeneration. Nicotinamide nucleoside (NR) is a metabolite found or produced in Saccharomyces cerevisiae. Nicotinamide nucleoside is a precursor to nicotinamide adenine dinucleotide (NAD+). [2]
In Alzheimer's disease models, nicotinamide nucleoside is thought to exert a protective effect by upregulating the expression of PGC-1α. Elevated PGC-1α levels promote the ubiquitination of BACE1 (a key enzyme involved in β-amyloid formation) and its subsequent proteasome degradation. This leads to a decrease in Aβ levels in the brain. [2]
Furthermore, nicotinamide nucleoside treatment upregulates the expression of a range of genes related to mitochondrial energy metabolism, which may help improve cognitive function and synaptic plasticity. [2]
This study suggests that nicotinamide nucleoside may be a potential strategy for treating Alzheimer's disease by targeting the PGC-1α-mediated pathway to reduce amyloid pathology and improve brain energy metabolism. [2]

C₁₁H₁₅N₂O₅

255.25

255.097

1341-23-7

Nicotinamide riboside chloride;23111-00-4;Nicotinamide riboside tartrate;2415657-86-0;Nicotinamide riboside malate;2415659-01-5

439924

Typically exists as solid at room temperature

1.201g/cm3

353.7ºC at 760mmHg

181.9ºC

3.05E-07mmHg at 25°C

1.549

-2.3

4

5

3

18

314

4

C1=CC(=C[N+](=C1)[C@H]2[C@@H]([C@@H]([C@H](O2)CO)O)O)C(=O)N

JLEBZPBDRKPWTD-TURQNECASA-O

InChI=1S/C11H14N2O5/c12-10(17)6-2-1-3-13(4-6)11-9(16)8(15)7(5-14)18-11/h1-4,7-9,11,14-16H,5H2,(H-,12,17)/p+1/t7-,8-,9-,11-/m1/s1

1-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]pyridin-1-ium-3-carboxamide

N-RibosylnicotinamideNicotinamide-beta-riboside Nicotinamide ribonucleoside

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.)