5-Amino 1MQ

5-Amino 1MQ (5-amino-1-methylquinolinium, CAS 42464-96-0) is a synthetic small molecule classified as a methylquinolinium derivative. [1] It was developed by Dr. Stanley J. Watowich’s research group at the University of Texas Medical Branch (UTMB) at Galveston as a selective inhibitor of the metabolic enzyme nicotinamide N-methyltransferase (NNMT). [2]

5-Amino 1MQ is an analogue of the parent molecule 1-methylquinolinium (1-MQ), modified with a primary amine substitution at the 5-position of the quinoline ring. This structural modification was rationally designed to optimize binding affinity to NNMT while dramatically improving membrane permeability — a critical limitation of the parent 1-MQ molecule. [3]

In conditions such as obesity, type 2 diabetes, and aging, NNMT is overexpressed in adipose tissue and skeletal muscle, where it depletes cellular pools of NAD+ and SAM. By blocking NNMT, 5-Amino 1MQ preserves these critical metabolic cofactors, shifting cellular metabolism from fat storage to fat oxidation and energy expenditure. [4]

Regulatory Status:

• FDA: NOT registered for human use — experimental research chemical. [5]
• WADA: Banned under S0 category (Non-Approved Substances) — prohibited at all times.
• GRAS: Not classified as Generally Recognized as Well-tolerated for dietary supplementation.

Developer: Ridgeline Therapeutics (Houston, TX), founded by Dr. Watowich, is advancing a lead NNMT inhibitor candidate (RT-002) toward Phase 1 first-in-human clinical trials, with IND-enabling GLP toxicology studies in minipigs currently underway. [6]

Pharmacokinetic Highlights:

• Oral Bioavailability: 38.4% (rats) [7]
• Half-Life: ~6.9 hours (oral), ~3.8 hours (IV) in rats
• Cmax: 2,252 ng/mL (oral, rats)
• Membrane Permeability: High (passive and active transport)

1. Primary Target — NNMT Enzyme

The molecular target of 5-Amino 1MQ is the cytosolic enzyme nicotinamide N-methyltransferase (NNMT), a metabolic regulator highly expressed in adipose tissue, liver, and skeletal muscle — particularly in obesity and type 2 diabetes. [2]

Binding Mechanism: 5-Amino 1MQ is a substrate-competitive inhibitor. It competes with nicotinamide (NAM) for the active binding site of NNMT, preventing the enzyme from catalyzing the transfer of a methyl group from SAM to NAM. This blockade prevents formation of 1-methylnicotinamide (1-MNA) and S-adenosyl-L-homocysteine (SAH). [3]

Potency:

• IC₅₀: ~1.0–1.2 µM for NNMT inhibition [1]
• EC₅₀: 2.3 ± 1.1 µM (reduction of intracellular 1-MNA levels) [1]
• Lipogenesis Inhibition: 30 µM reduced lipogenesis by 50%; 60 µM by 70%

2. Downstream Cascade A — NAD+ Salvage and SIRT1 Activation

NNMT normally acts as a “sink” for nicotinamide, permanently removing it from the NAD+ salvage pathway by methylating it into 1-MNA (which is then excreted). By inhibiting NNMT, 5-Amino 1MQ preserves the intracellular NAM pool, shunting it back into the NAD+ salvage pathway and significantly increasing intracellular NAD+ levels. [4]

Elevated NAD+ acts as a co-substrate for sirtuins, specifically activating SIRT1 — often called the “longevity gene.” SIRT1 activation drives increased mitochondrial biogenesis and metabolic rate. [8]

3. Downstream Cascade B — Methionine-SAM Cycle and Epigenetic Regulation

NNMT consumes SAM (the universal methyl donor) during NAM methylation. 5-Amino 1MQ prevents this consumption, increasing intracellular SAM levels and reducing SAH (a methylation inhibitor). This alters the cell’s epigenetic methylation potential, influencing histone and DNA methylation states that regulate gene expression for adipogenesis and metabolism. [4] [9]

4. Downstream Cascade C — Exercise Mimicry (Muscle-Specific)

In skeletal muscle, 5-Amino 1MQ triggers unique signaling: [10]

• Ribosomal Biogenesis: Upregulates proteins involved in ribosomal RNA biogenesis and aminoacyl-tRNA ligase activity, mimicking the protein translation signaling normally induced by exercise.
• Transsulfuration Pathway: Uniquely upregulates the transsulfuration pathway (via cystathionine β-synthase), enhancing protection against reactive oxygen species (ROS) via glutathione synthesis.
• AMPK Activation: Shifts the metabolome of sedentary muscle toward an exercised state via increased AMP, driving AMPK activation — a critical energy sensor promoting muscle hypertrophy and protein translation.

5. Receptor Selectivity

5-Amino 1MQ demonstrates high selectivity for NNMT, avoiding off-target effects: [3]

• Does NOT inhibit structurally related SAM-dependent methyltransferases: DNMT1, PRMT3, COMT
• Does NOT inhibit NAD+ pathway enzymes: NAMPT, SIRT1

This confirms that NAD+ and SAM increases result solely from preventing their degradation by NNMT, not from interfering with their synthesis or utilization enzymes.

6. Cellular and Tissue-Level Effects

Adipose Tissue (White Fat):

• Suppresses lipogenesis (fat creation) in adipocytes [1]
• Reduces white adipocyte size by >30% and WAT mass by ~35% [2]
• Produces a unique metabolomic signature (increased ketogenic amino acids)
• No alteration in food intake [2]

Skeletal Muscle:

• Activates senescent muscle stem cells (MuSCs), promoting proliferation and myofiber repair [11]
• Nearly 2-fold increase in myofiber cross-sectional area; ~70% increase in peak torque
• Sustained running capacity without fatigue taper [10]

Liver:

• Reverses hepatic steatosis (fatty liver) and normalizes ALT/AST [12]
• Reduces total plasma cholesterol by ~30% [2]

7. Pharmacokinetics

Parameter	Rat (Oral)	Rat (IV)
Oral Bioavailability	38.4%	—
Half-Life (T½)	6.9 ± 1.2 h	3.8 ± 1.1 h
Cmax	2,252 ng/mL	—
Membrane Permeability	High (passive + active transport)
Tissue Distribution	Adipose, muscle, liver; no 24h accumulation in heart/kidney/brain

Source: Awosemo/Neelakantan et al., J. Pharm. Biomed. Anal., 2021 [7]

🏋️ Obesity & Fat Loss

In diet-induced obese (DIO) mice, 5-Amino 1MQ (20 mg/kg SC, 3× daily, 11 days) reduced body weight by 5.1%, decreased epididymal white adipose tissue (WAT) mass by ~35% (P<0.001), and reduced adipocyte size by >30%, all without altering food intake. Plasma total cholesterol decreased ~30% (P<0.05). [2] In a longer study (32 mg/kg daily, 7 weeks), fat mass decreased by 29.3%, normalizing body composition to levels indistinguishable from age-matched lean controls. [9]

See also: AOD-9604 for related fat metabolism research.

💊 Type 2 Diabetes & Metabolic Syndrome

5-Amino 1MQ improved oral glucose tolerance and suppressed hyperinsulinemia in obese mouse models. It addresses the underlying metabolic dysfunction in white adipose tissue that drives insulin resistance. [12]

See also: Tirzepatide for related metabolic research.

💪 Muscle Regeneration & Sarcopenia

In aged (22-month) mice, 5-Amino 1MQ (10 mg/kg daily, 8 weeks) mimicked exercise effects: sedentary treated mice showed ~40% greater grip strength than untreated controls (P<0.001). Combined with exercise, grip strength increased by ~60%. Treated mice maintained a 1.8 km/day running increase at week 8, while untreated mice tapered off (P=0.0039). Intramyocellular lipid content decreased >30%. [10]

In aged (24-month) mice with acute muscle injury, treated animals showed nearly 2× greater myofiber cross-sectional area and ~70% increased peak torque (P<0.05), with enhanced muscle stem cell (MuSC) proliferation and fusion. [11]

🫁 Liver Disease (NAFLD/NASH)

Combined diet switch + 5-Amino 1MQ research application (28 days) normalized ALT and AST liver enzyme levels, reduced liver weight/size and triglyceride content, attenuated hepatic steatosis and macrophage infiltration. [12]

🧬 Duchenne Muscular Dystrophy (DMD)

Preclinical studies indicate that NNMT inhibition can improve muscle regeneration and function in DMD models by enhancing mitochondrial bioenergetics and reactivating dysfunctional muscle stem cells. [11]

🪸 Chronic Kidney Disease (CKD)

Research targeting NNMT inhibition shows potential in reducing renal fibrosis and tubular senescence, with improved kidney function and slowed disease progression in CKD models. [13]

🎯 Cancer (NNMT Overexpression)

NNMT is overexpressed in aggressive cancers including glioblastoma, ovarian cancer, and gastric cancer, driving metabolic and epigenetic remodeling that supports tumor growth. 5-Amino 1MQ is being investigated for its potential to suppress tumorigenesis, metastasis, and chemoresistance. [14]

⏳ Longevity & Anti-Aging

By elevating intracellular NAD+ levels and activating SIRT1, 5-Amino 1MQ is explored as a experimental to delay cellular aging, improve mitochondrial health, and prevent age-related physiological decline. [8]

Preclinical Animal Studies

• Obesity — DIO Mice (Neelakantan 2018): 20 mg/kg SC 3×/day, 11 days. -5.1% body weight (P<0.0001 at day 10), -35% WAT mass (P<0.001), >30% decreased adipocyte size and >40% decreased volume (P<0.05), ~30% lower plasma cholesterol (P<0.05). No change in food intake; no adverse toxicity. [2]
• Obesity + Diet (Sampson 2021): 32 mg/kg SC daily, ~7 weeks (DIO mice switched to lean diet). -29.3% fat mass from baseline vs. -2.9% for diet alone. Body composition normalized to lean controls. Metabolomic analysis predicted lipid synthesis inhibition (z-score = -2.566, P=0.045). [9]
• Microbiome (Dimet-Wiley 2022): 32 mg/kg SC daily, ~7 weeks. Treated mice showed distinct microbiome cluster with increased Lactobacillus and Parasutterella; decreased Erysipelatoclostridium. [15]
• Metabolic/Liver (Babula 2024): Daily SC, 28 days (DIO mice). Normalized ALT/AST, improved oral glucose tolerance, suppressed hyperinsulinemia, reduced liver weight and triglycerides, attenuated steatosis and macrophage infiltration. [12]
• Exercise Mimicry (Dimet-Wiley 2024): 10 mg/kg SC daily, 8 weeks (aged 22-month mice). Sedentary treated: +40% grip strength (P<0.001). Exercise + treated: +60% grip strength, maintained 1.8 km/day running increase at week 8 (P=0.0039). >30% reduction in intramyocellular lipid. [10]
• Muscle Regeneration (Neelakantan 2019): 5–10 mg/kg, 1–3 weeks (aged 24-month mice with acute injury). ~2× myofiber CSA, +70% peak torque (P<0.05), increased MuSC proliferation and fusion. [11]
• Peripheral Artery Disease (Dong 2025): Daily dosing (BALB/cJ mice with hindlimb ischemia). Significantly improved muscle strength (P<0.0001), power (P=0.031), total work (P=0.037). Independent of perfusion or capillary changes. [16]

Pharmacokinetic Profile (Rats)

Awosemo/Neelakantan et al. (2021): Oral bioavailability 38.4%; T½ 6.9h (oral) / 3.8h (IV); Cmax 2,252 ng/mL. High membrane permeability. No 24-hour accumulation in heart, liver, kidney, or brain (recirculation noted at ~12h). Cross-species liver metabolic stability confirmed. [7]

reported tolerability profile (Preclinical)

Acute Toxicity: In mice, animals survived doses from 50 mg/kg to 2,000–5,000 mg/kg with no observable adverse reactions during 48-hour monitoring. [2]

Subacute Toxicity (14 days): Liver (AST, GGT), heart (troponin I), and inflammatory (CRP) markers were unaffected except for significant CRP rise at highest IV dose (200 mg/kg) at 6 hours post-dose. [2]

Cell Viability: No impact up to 100 µM; modest cytotoxicity at 100–300 µM; ~40% cytotoxicity at 600 µM in 3T3-L1 adipocytes. [1]

Clinical Development Status

Ridgeline Therapeutics (founded by Dr. Watowich) is developing a lead NNMT inhibitor candidate (RT-002), conducting IND-enabling GLP toxicology studies in minipigs with the goal of submitting an IND briefing package to the FDA for Phase 1 first-in-human clinical trials. [6]