NNMTi is a potent inhibitor of nicotinamide N-methyltransferase (NNMT) (IC50=1.2 μM) that selectively binds to NNMT substrate binding site residues. NNMTi promotes myoblast differentiation in vitro and enhances the fusion and regeneration capacity of muscle stem cells in aged mice.

NNMTi

NNMTi has been implicated in various metabolic and pathological processes, including obesity, cancer, diabetes, and neurological disorders, making its inhibitors critical for therapeutic development.

NNMT FUNCTION

The discovery of NNMTi stems from efforts to target dysregulated metabolic and aberrant methylation pathways. Elevated NNMT activity has been observed in many cancers, including breast, pancreatic, and lung cancers, where it contributes to tumor growth and resistance to apoptosis. Similarly, overexpression of NNMT in adipose tissue is associated with obesity-related insulin resistance. Recognizing these associations, researchers began developing NNMT inhibitors to restore normal metabolic balance and slow disease progression.

In Vitro

NNMTi (10-30 μM; 96 hours) produced a concentration-dependent increase in myoblast differentiation in C2C12 myoblasts. 30 μM NNMTi produced 18% MHC-positive myotube nuclei, which was a 45% increase in myoblast differentiation compared to untreated differentiating myoblasts (12% MHC-positive myotube nuclei).

DMSO : 20.83 mg/mL (72.80 mM; ultrasonic and warming and heat to 60°C)

H2O : 2.27 mg/mL (7.93 mM; ultrasonic and warming and heat to 60°C)

NNMTi TEST 1

In Vivo

NNMTi (subcutaneous injection; 5 mg/kg and 10 mg/kg; 2 weeks, 1 week before injury and 1 week after injury) had an effect on post-injury muscle regeneration, increasing the incidence of proliferating/active muSCs by 60% and 75% at 5 mg/kg and 10 mg/kg, respectively. The relative number of fibers with EdU+ myonuclei increased by 40% and 48% at 5 mg/kg and 10 mg/kg of NNMTi treatment, respectively. The odds ratio of control fused myonuclei was 0.58 and 0.53 times that of the low and high doses of NNMTi, respectively.
NNMTi (subcutaneous injection; 10 mg/kg; 1 week) did not produce systemic toxicity in mice, and glucose, cholesterol, plasma protein, and electrolyte levels did not differ between control and NNMTi-treated samples in mice. Repeated daily administration of NNMTi for 1 week after injury had no adverse systemic toxicity or behavioral effects in aged mice.

NNMTi TEST 2

1. Inhibit tumor proliferation and enhance drug efficacy
In oncology, NNMT inhibitors have shown the potential to inhibit tumor proliferation and enhance the efficacy of existing chemotherapy drugs in preclinical studies. By disrupting the NNMT-mediated methylation pathway, these inhibitors impair energy metabolism and epigenetic adaptation of cancer cells.

2. Improve insulin sensitivity
In terms of metabolic disorders, NNMTi compounds have been studied for their ability to improve insulin sensitivity and promote weight loss, thereby addressing the root causes of type 2 diabetes and obesity.

3.Expected to delay neurodegeneration
It is emerging as a promising candidate drug in neurodegenerative disease research. NNMT has been implicated in the pathogenesis of Alzheimer's and Parkinson's diseases by affecting cellular methylation status and oxidative stress. By regulating NNMT activity, researchers hope to develop new interventions to slow neurodegeneration and enhance neuronal resilience.

4.Expected to support personalized treatment plans in the future
The development of NNMT inhibitors has also stimulated interest in diagnostic applications. Elevated NNMT levels in biological fluids are considered potential biomarkers for cancer and metabolic disorders. Therefore, NNMTi compounds could play a role in therapeutic and diagnostic strategies, supporting a more personalized approach to treatment.

Small molecule products are mostly prepared with DMSO or water. If sterilization is required, it is recommended to filter the diluted working solution with a 0.22 μm sterile filter membrane for sterilization, and ensure that both the mother solution and the working solution are clear before filtering, and no precipitation is observed under the microscope. Do not sterilize at high temperature or high pressure.

1. If DMSO is used as the solvent, since DMSO itself has good bactericidal power, the prepared solution is generally a sterile solution. It is only necessary to keep the operating environment and instruments sterile. It is recommended to prepare 1000×DMSO mother solution before the experiment. If contamination occurs, filter and sterilize the working solution.
2. If water is used as the solvent, it is recommended to prepare a mother solution of more than 50× with sterile water, dilute it to the working solution, and then filter and sterilize it.

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