SANA (MVD1): A Nitroalkene Salicylate and the Creatine-Dependent Thermogenesis Pathway

SANA (MVD1) is an investigational small molecule that has drawn attention for a mechanism almost nothing else in metabolic pharmacology shares: it appears to raise energy expenditure in adipose tissue through creatine metabolism rather than by acting on appetite. Chemically it is 5-(2-nitroethenyl)salicylic acid — a nitroalkene grafted onto the salicylate scaffold that gave us aspirin. Eolo Pharma developed the compound and assigned it the clinical code MVD1, and in June 2025 Nature Metabolism published its full preclinical characterization alongside a first-in-human Phase 1A/B trial [1].

This article covers what the published record actually shows: the chemistry, the mechanism and its open questions, the trial data, and — importantly for anyone sourcing the compound — the specific analytical problems that make SANA harder to verify than its simple structure suggests. SANA (MVD1) is supplied strictly for laboratory research use.

Chemical Identity of SANA (MVD1)

The structure is deceptively simple. Salicylic acid supplies the aromatic core, and a nitroethenyl (–CH=CH–NO₂) group occupies the 5-position. That single substituent does all the interesting work.

Why the Nitroalkene Group Matters

Nitroalkenes are electrophilic. They undergo reversible Michael addition with thiol groups, which lets them modify cysteine residues on proteins and shift signalling in ways the parent scaffold cannot [1]. Researchers had already moved this reactive group onto other scaffolds — a vitamin E analogue, a benzoic acid derivative — and observed that the scaffold shapes where the nitroalkene acts while the nitroalkene supplies the reactivity.

SANA is the salicylate version of that strategy, and the structure–activity data are unusually clean. The published work tested four close relatives:

• M1, the saturated metabolite 5-(2-nitroethyl)salicylic acid, formed in vivo — inactive on diet-induced obesity and glucose handling
• 3-SANA, the nitroalkene at the 3-position — inactive on thermogenesis
• 4-SANA, the nitroalkene at the 4-position — inactive on thermogenesis
• E-SANA, an extended-chain 5-(2-nitroallyl) variant — inactive on thermogenesis

Only the 5-nitroethenyl arrangement activated thermogenesis or induced creatine kinase expression in brown adipose tissue [1]. In other words, both the presence of the nitroalkene and its exact position on the ring are required. That finding matters enormously for sourcing, and we return to it below.

Mechanism: The Futile Creatine Cycle

To understand SANA (MVD1), you need the pathway it targets.

Classical non-shivering thermogenesis runs through uncoupling protein 1 (UCP1), which dissipates the mitochondrial proton gradient as heat. But a second, UCP1-independent route exists. In 2015, Kazak and colleagues described a creatine-driven substrate cycle in beige fat, in which creatine phosphorylation and phosphocreatine hydrolysis run simultaneously — burning ATP and releasing heat without any net work [2]. Later work identified creatine kinase B (CKB) as a controller of this futile cycle in thermogenic fat [3], and subsequent studies confirmed the cycle can power thermogenesis in classical brown adipose tissue independently of UCP1 [4].

SANA appears to switch this pathway on pharmacologically. The evidence in the Nature Metabolism paper is layered:

Proteomics pointed the way. Unbiased proteomic analysis of inguinal white adipose tissue from treated mice showed enrichment of catabolism, oxidative phosphorylation and thermogenesis pathways. Glycine amidinotransferase (GATM) — the rate-limiting enzyme in creatine synthesis — appeared only in the treated group [1].

Creatine levels rose. Mass spectrometry confirmed increased total creatine in both inguinal white and brown adipose tissue [1].

Gene expression followed. Treatment upregulated Ckmt1 and Ckm in white adipose tissue, Ckmt2 and Ckm in brown adipose tissue, plus creatine synthesis and transport genes Gatm, Oat, Prodh and Slc6a8 [1].

Knockouts isolated the pathway. SANA still drove weight and fat mass loss in UCP1-knockout mice, and still protected AMPK(α1)-knockout mice from diet-induced obesity — ruling out both conventional mechanisms [1].

Creatine depletion abolished the effect. Treating mice with β-guanidinopropionic acid, a creatine antagonist, eliminated the thermogenic response entirely [1].

That last experiment is the cleanest piece of evidence. Remove creatine, and the compound stops working.

What the Preclinical Data Showed

Across rodent models of diet-induced obesity, the published findings were consistent [1]:

• Mice on a high-fat diet with SANA were substantially protected from weight gain, at doses where the parent salicylate was completely ineffective
• Food intake, stool mass, caloric content of stool, digested energy and physical activity were all unchanged — so the effect was not appetite suppression or malabsorption
• Fat mass fell across every adipose depot measured, and lean mass rose as a percentage of body composition
• Livers of treated obese mice were macroscopically and histologically normal, with transaminases protected
• Glucose tolerance, fasting insulin, insulin sensitivity and HOMA-IR all improved
• In mice already obese before treatment began, SANA produced actual weight loss and outperformed both salicylate and metformin on glycaemic control over three weeks
• Normal-chow mice showed no weight change and no change in energy expenditure

One caveat worth stating precisely, because it is often overstated elsewhere: the paper reports an increase in the percentage of lean mass. That reflects preferential loss of fat, not demonstrated accretion of new lean tissue. Aerobic exercise capacity, electromyographic shivering response, skeletal muscle creatine kinase expression and cardiac mitochondrial respiration were all unaffected [1].

The thermoneutrality problem

A standing objection to thermogenesis-based approaches is that laboratory mice live cold-stressed at 22 °C, while humans spend most of their lives near thermoneutrality — so thermogenic effects in mice may not translate.

The SANA work addressed this directly. Under thermoneutral housing at 28 °C, the compound still protected against diet-induced obesity, hyperglycaemia and glucose intolerance. In a short acute feeding protocol where body weight had not yet diverged between groups, thermal imaging showed increased heat dissipation and indirect calorimetry showed a genuine rise in energy expenditure [1]. Because body weight was matched, that increase cannot be dismissed as a downstream consequence of being leaner.

The Phase 1A/B Human Trial

Eolo Pharma ran a randomized, double-blind, placebo-controlled first-in-human trial, registered with the Australian New Zealand Clinical Trials Registry as ACTRN12622001519741 [1]. The compound was referred to as MVD1 throughout clinical development.

Design. A single ascending dose (SAD) arm enrolled 17 healthy lean volunteers across 200–800 mg. A multiple ascending dose (MAD) arm enrolled 24 volunteers with overweight or obesity at 200–400 mg per day for 15 days, conducted as an inpatient study with a high-carbohydrate diet provided ad libitum. The primary endpoint was safety; secondary endpoints covered tolerability and pharmacokinetics; body weight, glucose and insulin were exploratory.

Safety. Two participants experienced adverse events judged definitively drug-related — reversible renal tubular damage evidenced by proteinuria and glucosuria — both at the highest single dose (800 mg). No definitively drug-related adverse events occurred during the multiple-dose phase. The most frequent possibly-related events were headache and soft stools. No severe adverse events were reported, and liver function was normal in all participants after 15 days. Notably, no moderate or severe gastrointestinal adverse effects were recorded [1].

Pharmacokinetics. Profiles resembled those seen in mice. Day 15 exposure was comparable to day 1, indicating the compound does not accumulate [1].

Exploratory findings. In the highest MAD cohort (200 mg every 12 hours), participants showed a significant decrease in body weight over 15 days — roughly 3%, which the authors note is comparable to semaglutide over the same interval. Plasma glucose, insulin, HOMA-IR and fructosamine all improved. Placebo participants showed no such changes [1].

The limitation the authors themselves stress. The trial was designed and powered for safety, not efficacy, with a small number of volunteers and limited measurements. The efficacy signals are preliminary and require a Phase 2 trial with longer treatment, larger samples and extended endpoints before they can be considered validated [1].

Where the Mechanism Is Still Contested

Honest coverage requires acknowledging that the futile creatine cycle is an actively debated field, not settled biology.

Nicholls and Brand published a critical assessment questioning whether creatine cycling contributes meaningfully to brown adipose tissue thermogenesis on bioenergetic grounds [5]. The identity of the controlling kinase also remains unresolved: CKB is the only isoform genetically demonstrated to mediate creatine kinase activity in mouse adipocyte mitochondria [3,4], yet SANA upregulated Ckmt1, Ckm and Ckmt2 — and notably did not affect Ckb, even though cold exposure does [1]. There are species differences too, with CKB reported as exclusively cytosolic in human adipose cells but mitochondrial in mouse adipocytes.

The authors are candid about this. Their own reading is that SANA does not fully phenocopy cold-elicited creatine-dependent thermogenesis, and may work through different kinases. They also state plainly that the molecular target of SANA remains undetermined [1].

One safety-relevant observation deserves attention: Ckmt1-knockout mice treated with SANA showed roughly 50% mortality within a week at standard housing temperature, rescued entirely by thermoneutral housing [1]. The interpretation offered is that the compound may commit animals to creatine-dependent thermoregulation at the expense of thermogenic flexibility. That is a genuine mechanistic finding, and any research design involving creatine-pathway-compromised models should account for it.

SANA Compared With the Salicylate Scaffold

Salicylate itself has metabolic activity. It activates AMPK [6], and salsalate improved glycaemic control in randomized trials in type 2 diabetes [7]. It can also uncouple mitochondria directly [8]. But it requires very high doses, and in the SANA experiments the parent compound was ineffective at doses where SANA produced maximal effects [1].

The mechanistic separation is sharp. Salicylate’s thermogenic contribution involves direct mitochondrial uncoupling; SANA does not uncouple mitochondria directly. Adding SANA to isolated mitochondria from adipose tissue or brain produced no effect on respiration at all, and neither did direct treatment of adipocytes — the respiratory increase only appeared after sustained in vivo exposure, pointing to a genomic rather than acute mechanism [1].

Analytical Considerations: Why SANA Is Harder to Verify Than It Looks

This is where SANA (MVD1) becomes an unusually demanding compound to authenticate, and it follows directly from the structure–activity data above.

Mass spectrometry alone cannot confirm identity. The inactive regioisomers 3-SANA and 4-SANA share SANA’s molecular formula (C₉H₇NO₅) and its monoisotopic mass (209.0324 Da) exactly. They are isobaric. No MS method — nominal or high-resolution — distinguishes them on mass. A certificate showing the correct molecular ion is entirely compatible with a batch of biologically inactive regioisomer.

Alkene geometry is also mass-silent. The active compound is the E (trans) isomer. A Z-configured batch would be indistinguishable by mass and difficult to catch by reversed-phase HPLC without an authenticated standard.

What MS does catch. The saturated metabolite M1, 5-(2-nitroethyl)salicylic acid, is C₉H₉NO₅ at 211.048 Da — a +2.02 Da shift that mass spectrometry resolves cleanly. M1 was confirmed inactive in vivo [1], so this is a meaningful check, just not a sufficient one.

The method that actually resolves it is NMR. Aromatic substitution patterns distinguish the 3-, 4- and 5-regioisomers through their coupling patterns, and vinyl proton coupling constants separate E from Z geometry. For this compound class, ¹H NMR is not a supplementary technique — it is the primary identity method, with HPLC quantifying purity and MS confirming molecular mass. This is the orthogonal-methods logic that runs through our whole third-party COA verification programme, and SANA is close to a textbook case for it.

Handling implications of the nitroalkene. Because the nitroalkene is an electrophilic Michael acceptor that reacts with low-molecular-weight thiols [1], thiol-containing buffer components — dithiothreitol, β-mercaptoethanol, high free glutathione — will quench it. Design in vitro work accordingly. The published formulation work used pH-controlled solubilization in phosphate buffer around pH 6.5, and DMSO stocks are standard for cell work. Store cold, desiccated and protected from light.

Class Context

SANA (MVD1) sits in a small group of research compounds aimed at energy expenditure rather than appetite. SLU-PP-332, an ERR pan-agonist studied as an exercise mimetic, targets oxidative capacity through a transcriptional route. 5-Amino-1MQ, an NNMT inhibitor, is investigated for effects on adipocyte NAD⁺ salvage and cellular energetics. Each addresses a different node, and none shares SANA’s creatine-cycle mechanism. Further reading on this compound class is collected under metabolic compounds.

The framing the authors themselves use is complementarity rather than competition: incretin-based agents act primarily on intake, while SANA acts on expenditure, and the two axes are not mutually exclusive [1].

Regulatory Status

SANA (MVD1) is an investigational compound in early clinical development. It has no approved brand name, no marketing authorization in any jurisdiction, and no FDA approval for human use. It remains the proprietary lead candidate of Eolo Pharma. Kimera Chems supplies it strictly for in vitro and laboratory research use only — not for human or veterinary use, ingestion, injection or topical application.

Frequently Asked Questions

What is SANA (MVD1)? An investigational nitroalkene derivative of salicylate, 5-(2-nitroethenyl)salicylic acid, studied as a research tool for creatine-dependent thermogenesis and adipose tissue energy metabolism.

How does SANA work? Published data indicate it activates non-shivering thermogenesis in brown and white adipose tissue through a creatine-dependent pathway, independent of UCP1 and AMPK. Creatine depletion abolishes the effect. The direct molecular target has not yet been identified.

Why is SANA also called MVD1? MVD1 is the clinical development code Eolo Pharma assigned to the compound. Both names refer to the same molecule.

Is SANA approved for human use? No. It has completed a Phase 1A/B safety trial and remains investigational.

Why does the isomer matter so much? Because the 3- and 4-position regioisomers and the saturated metabolite were all shown to be biologically inactive. Since the regioisomers are isobaric with SANA, mass spectrometry cannot rule them out — NMR is required for confident identity confirmation.

References

1. Cal K, Leyva A, Rodríguez-Duarte J, et al. A nitroalkene derivative of salicylate, SANA, induces creatine-dependent thermogenesis and promotes weight loss. Nature Metabolism. 2025;7(8):1550–1569. doi:10.1038/s42255-025-01311-z. https://www.nature.com/articles/s42255-025-01311-z
2. Kazak L, Chouchani ET, Jedrychowski MP, et al. A creatine-driven substrate cycle enhances energy expenditure and thermogenesis in beige fat. Cell. 2015;163(3):643–655. doi:10.1016/j.cell.2015.09.035
3. Rahbani JF, Roesler A, Hussain MF, et al. Creatine kinase B controls futile creatine cycling in thermogenic fat. Nature. 2021;590(7846):480–485. doi:10.1038/s41586-021-03221-y. https://www.nature.com/articles/s41586-021-03221-y
4. Bunk J, Ersin M, Hussain MF, et al. The futile creatine cycle powers UCP1-independent thermogenesis in classical brown adipose tissue. Nature Communications. 2025. doi:10.1038/s41467-025-58294-4. https://www.nature.com/articles/s41467-025-58294-4
5. Nicholls DG, Brand MD. A critical assessment of the role of creatine in brown adipose tissue thermogenesis. Nature Metabolism. 2023;5:21–28. https://www.nature.com/articles/s42255-022-00718-2
6. Hawley SA, Fullerton MD, Ross FA, et al. The ancient drug salicylate directly activates AMP-activated protein kinase. Science. 2012;336(6083):918–922.
7. Goldfine AB, Fonseca V, Jablonski KA, et al. The effects of salsalate on glycemic control in patients with type 2 diabetes: a randomized trial. Annals of Internal Medicine. 2010;152(6):346–357.
8. Smith BK, Ford RJ, Desjardins EM, et al. Salsalate (salicylate) uncouples mitochondria, improves glucose homeostasis, and reduces liver lipids independent of AMPK-β1. Diabetes. 2016;65(11):3352–3361.
9. National Center for Biotechnology Information. PubChem Compound Summary for CID 54133300, 2-Hydroxy-5-(2-nitroethenyl)benzoic acid. https://pubchem.ncbi.nlm.nih.gov/compound/54133300

SANA (MVD1) is sold strictly for in vitro research and laboratory use only. It is not a drug, supplement, food or cosmetic, and is not intended for human or veterinary use, ingestion, injection or topical application. Batch-specific analytical data is available in the COA archive.