date: 2021-10-07 00:12

Caswell No. 392; Sulfo Black B; Nitro Kleenup

DNP (2,4-dinitrophenol)

protonophore; mitochondrial uncoupler of phosphorylative oxidation and ATPase.

During oxidative phosphorylation, DNP's primary target of action is on the *proton pump*, where the mitochondrial matrix actively (endergonically) pumps protons into the intermembrance space- DNP "sabotages" this by allowing the protons to leak back into the mitochondrial matrix. Rather than permitting potential energy to form via an electrochemical gradient of protons with a lower concentration in the mitochondrial matrix versus the intermembrane space (natural state), a higher concentration build in the mitochondrial matrix and energy must be dissipated as heat (increase in core temperature)._[1]

Decreases efficiency of (catabolic) energy metabolism ⇒ heat

• Complete nutrient catabolism is between 30 and 40 percent efficient, and some of the energy is therefore released as heat. Heat is a vital product of nutrient catabolism and is involved in maintaining body temperature. If cells were too efficient at transforming nutrient energy into ATP, humans would not last to the next meal, as they would die of hypothermia.

Additional aspects of DNP's mechanisms:

• Ca²⁺ release from mitochondria & SR into the cytosol -- which subsequently must be pumped back (caused by effects on membrane potential from oxidative phosphorylation)_[6]
• Interferes with Pᵢ groups' uptake into the mitochondrial matrix, thereby reducing substrates for ATPase_[7]

dose-response dose-response: average BMR ↑ 11% * 100mg⁻¹_[3][12]... but varies widely between individuals_[12] (in some cases, DNP alone, without dietary modifications, results in no significant weight loss)_[13]

• 4 mg/kg results in ~40% increase to RMR (mean)

The method employed consisted of the administration of one capsule (100 mg of sodium dinitrophenol) the first day, two capsules the second day, and three capsules daily thereafter (preferably after meals)... Thereafter the dose was regulated by...[A] rule was followed invariably that, when the temperature rose as high as 99.2° F (37.33° C) [indicative of acute toxicity], the drug was either discontinued or the dosage sharply reduced..._[12]

With long-term administration (> 7 weeks), two factors lead to sudden cessation of weight loss (after weeks of steady, rapid weight loss):

1. Factor 1: rapidly acquired tolerance to the drug. EX:
   • One patient, on a daily dose of 320 mg, resulting in a RMR ↑26%, lost weight at an average rate of 1.4 kg (3 lb) weekly x 7 weeks. Then, for 6 weeks, she lost no weight even though the dose was raised to 400 mg daily. A RMR determination at this time showed that RMR ↑ by a mere 6% (relatively rare)
   • The acquired tolerance is rapidly lost if the drug is discontinued for as short a period as two weeks. The dinitrophenol may then be resumed at a lower dosage level with its original effect on the patient.
2. Factor 2: Edema (profound); DNP promotes water storage in the tissues of the body. EX:
   • Another patient, on a daily dose of 300 mg, resulting in a RMR ↑46%, lost an average of 0.9 kg (2 lb) weekly x 8 weeks. Then, she suddenly stopped losing weight. The dosage was increased to 400 mg daily x 4 weeks with no resultant weight loss. A RMR determination at this time showed that RMR ↑ by a considerable 62% (common)
   • Naturally, cessation from DNP at such a time results in dramatic water, and therefore weight, loss.

_[12]

Seasonal phenomenon In climates subject to warm and cold months, a seasonal phenomenon is observed:

As summer passes into fall, it is found that the patient suddenly ceases to lose weight, although the weight loss before may have been very satisfactory and steady. That the mechanism involved is not the development of tolerance is attested by the fact that a considerable group of patients exhibit this phenomenon simultaneously. In the case of dinitrophenol it may be partly accounted for, but only partly, by the diminution of sweating that accompanies the colder weather. In the hot summer months, small doses of dinitrophenol will produce a weight loss that will be matched in the cold winter months only by considerably larger doses. _[12]

Myocardial effects; EKG changes

Changes that may occur (moderate likelihood), indicate per Taussig (56) the development of a "prolongation of the recovery time of the heart muscle together with a decrease in the voltage initiated by ventricular contraction and suggest that some alteration in the metabolism of the myocardium has resulted from dinitrophenol... There is no evidence that dinitrophenol causes death through a toxic action on the heart."_[12]

_{[8] [9] [10]}

DNP, by uncoupling of oxidative phosphorylation ⇒ ↑↑O₂ consumption (reflects ↑RMR) & ADP:ATP ratio, ↑lactate synthesis (the latter due to a compensatory ↑glycolysis due to low ATP, thereby ↑pyruvate [glycolysis end-product]) & ↓glucose (due to the ↑glycolysis)_[8]... therefore, hyperlactacidemia is a serious concern with chronic DNP use.

Utility of sodium bicarbonate?

Results: Blood bicarbonate and pH were elevated at all time points after ingestion in sodium bicarbonate (BIC) vs PLA (p < 0.05). The protein content of monocarboxylate transporter 1 (MCT1) and basigin (CD147), at 6 h and 24 h post-exercise, and sodium/hydrogen exchanger 1 (NHE1) 24 h post-exercise, were significantly greater in BIC compared to PLA (p < 0.05), whereas monocarboxylate transporter 4 (MCT4), sodium/bicarbonate cotransporter (NBC), and carbonic anhydrase isoform II (CAII) content was unchanged. These increases in protein content in BIC vs. PLA after acute sprint-interval exercise may be associated with altered physiological responses to exercise, such as the higher blood pH and bicarbonate concentration values, and lower exercise-induced oxidative stress observed during recovery (p < 0.05). Additionally, mitochondrial respiration decreased after 24 h of recovery in the BIC condition only, with no changes in oxidative protein content in either condition.

Concusion: Sodium bicarbonate ingestion induces metabolic alkalosis that in turn induces post-exercise increases in several lactate/pH regulatory proteins, and reveal an unexpected role for acidosis in mitigating the loss of mitochondrial respiration caused by exercise in the short term... Thomas C, Delfour-Peyrethon R, Lambert K, Granata C, Hobbs T, Hanon C, Bishop DJ. The effect of pre-exercise alkalosis on lactate/pH regulation and mitochondrial respiration following sprint-interval exercise in humans. Front Physiol. 2023 Jan 27;14:1073407. doi: 10.3389/fphys.2023.1073407. PMID: 36776968; PMCID: PMC9911540.

Sodium bicarbonate, then, may decrease efficacy of DNP (by ↓O₂ consumption, that reflects RMR)?

As a lactate buffering & metabolic alkalinising agent, may reduce symptoms of hyperlactacidemia.

But, from Rudnick MR, Blair GJ, Kuschner WG, Barr J. Lactic Acidosis and the Role of Sodium Bicarbonate: A Narrative Opinion. Shock. 2020 May;53(5):528-536. doi: 10.1097/SHK.0000000000001415. PMID: 31318832:

> Lactic acidosis occurs commonly and can be a marker of significant physiologic derangements. However what an elevated lactate level and acidemia connotes and what should be done about it is subject to inconsistent interpretations. This review examines the varied etiologies of lactic acidosis, the physiologic consequences, and the known effects of its treatment with sodium bicarbonate. Lactic acidosis is often assumed to be a marker of hypoperfusion, but it can also result from medications, organ dysfunction, and sepsis even in the absence of malperfusion. Acidemia causes deleterious effects in almost every organ system, but it can also have positive effects, increasing localized blood flow and oxygen delivery, as well as providing protection against hypoxic cellular injury. The use of sodium bicarbonate to correct severe acidemia may be tempting to clinicians, but previous studies have failed to show improved patient outcomes following bicarbonate administration. Bicarbonate use is known to decrease vasomotor tone, decrease myocardial contractility, and induce intracellular acidosis. This suggests that mild to moderate acidemia does not require correction. Most recently, a randomized control trial found a survival benefit in a subgroup of critically ill patients with serum pH levels <7.2 with concomitant acute kidney injury. There is no known benefit of correcting serum pH levels ≥ 7.2, and sparse evidence supports bicarbonate use <7.2. If administered, bicarbonate is best given as a slow IV infusion in the setting of adequate ventilation and calcium replacement to mitigate its untoward effects.

From [ProM post, Type-IIx, Sun Feb 19 2023](https://www.professionalmuscle.com/forums/index.php?threads/no-longer-feeling-dnp-work-towards-the-end.174114/post-3198727):

My initial thinking is that sodium bicarbonate ingestion presents a double-edged sword. It can reduce mitochondrial respiration (e.g., post-anaerobic exercise bout) & serve as a lactate buffering and metabolic alkaline agent to reduce symptoms of hyperlactacidemia. Since DNP by uncoupling of oxidative phosphorylation ⇒ ↑↑O₂ consumption (reflects ↑RMR) & ADP:ATP ratio, it ↑lactate synthesis (due to a compensatory ↑glycolysis due to low ATP). As such, it causes hyperventilation (highly ergolytic, or performance worsening) & hyperlactacidemia. Unfortunately, since hyperventilation is the sine qua non of the increase to RMR, if sodium bicarbonate ingestion reduces it to some extent, that degree of change basically reflects the degree of diminished efficacy...[truncated]...

AMPK activation Can fairly anticipate that the increased ADP/ATP ratio signals to activate AMPK (Metformin does as well by mitochondrial electron transport Complex I inhibition). AMPK activation inhibits mTORC1/P70S6K1 phophorylation, thereby inhibiting hypertrophy signals.

• See also: [Article on Metformin risk-reward profile](210901-1854.md) & its [AMPK](210901-1854.md#AMPK) section

Less practically useful as an antidiabetes agent than aspirin_[9].

Time-course dependent (bimodal effects) From _[12]:

Apparently the conclusions to be drawn are that, with short courses of dinitrophenol (e.g., 5 days) administration in normal individuals, there is moderate elevation of the fasting blood sugar (though not hyperglycemia) and a moderate impairment of carbohydrate tolerate (i.e., GTT), in the majority of cases. With prolonged administration (e.g., 21 - 420 days) there is no change in the fasting blood sugar and a marked increase of carbohydrate tolerance... With prolonged administration in cases of diabetes there is apparently an increase of carbohydrate tolerance.

No significant effect on thyroid function

> DNP is known to occupy thyroid binding proteins in blood. That leaves less of the binding protein to bind to thyroid, which can increase free thyroid levels. In the short term, that can result in higher serum levels of thyroid and accelerated clearance from the body. However, a new steady state is soon reached in which thyroid function will be normal, despite potentially reduced serum levels of total T4 and total T3. As one paper on DNP explained, "this action could lower the total hormone concentration in serum but should have no persistent effect on thyroid function". This means that even though the level of thyroid hormones in the blood can be lower than normal while on DNP, thyroid function will still be normal. So if you're seeing a doctor and having blood work done to determine your correct thyroid dose, you would not want to be taking DNP, since it could alter your numbers. Only after your dose has been set and no blood work is planned would you want to take DNP, during which time you'd stay on your thyroid medication. [Conciliator @ MesoRx Oct 9 2009](https://thinksteroids.com/community/threads/dnp-with-extant-hypothyroid.134279154/post-652067)

> It's not quite clear that DNP decreases T3 levels in practice. Theres a case study that's often posted as evidence for this, but I'm of the opinion that the bodybuilders in question had low thyroid levels not because of the DNP, but because they were taking T3 during their cycle. Exogenous T3 definitely will suppress endogenous production. I had a Norwegian translate the paper for me and it was unclear why they had low thyroid levels. They didn't test for reverse thyroid when they could have, which would have made the cause clearer. I also have a friend who's a doctor of internal medicine who ran 600mg/day for 3-4 months (went from 320 lbs down to the mid 200's). He gave himself frequent blood tests and said his thyroid didn't budge. I'm just not convinced that DNP lowers thyroid levels. Even if it did, you can simply bump up your DNP dosage to keep weight loss humming along. This is what they did in the original research after they found that there was a rapidly developing tolerance to the drug in the first couple of weeks. They just upped the dosage a little to overcome the tolerance. No one was given thyroid, even though subjects were kept on the drug for many months at a time to over a year. I like this option because thyroid is notoriously catabolic. I think it's generally best to just leave it alone. [Conciliator @ Steroid.com May 14 2007](https://forums.steroid.com/anabolic-steroids-questions-answers/300488-why-dnp-t3-together.html#post3478526)

Practically, my view is that if you're using DNP, the objective cannot be to build new muscle tissue (as ATP is going to be dramatically reduced: thus the energy state of the cell directs energy metabolism to catabolic processes).

As such, just use a - here we go again - minimal effective dose. So that means, don't go under 100 mg daily (no real justification; even at 100 - 200 mg daily, DNP is less efficacious as an insulin sensitizing/antidiabetic agent than aspirin in humans; and at 100 mg, expect a modest ~10% RMR increase [less in heavy bodybuilders]).

Certainly, a keto diet can rationally serve this objective: in an individual with a low b.f.%, with the use of MCTs, et cetera, keto can be used to really reduce those difficult to reduce fat depots like the lower back in men & the hips/thighs in women. DNP will make this all the more attainable (and we can "get in and get out," staving off longer-term muscle catabolism).

Theoretically the reason that DNP does lower blood glucose concentrations (but remember, it's a minor effect in humans) is by uncoupling of oxidative phosphorylation ⇒ ↑↑O₂ consumption (reflects the increase in RMR) & ADP:ATP ratio, ↑lactate synthesis (the latter due to a compensatory ↑glycolysis due to low ATP, thereby ↑pyruvate [gycolysis end-product]) & ↓glucose (due to the ↑glycolysis): hence hyperlactacidemia is a serious concern with DNP use.

So yes: with DNP, keto makes sense (i.e., entering ketosis); anticatabolic agents that are selective in the intracellular compartment (androgens) make sense as well; the utility of rhGH here is complicated by its hyperglycemic and insulin resistance effects, but it certainly makes sense for longer-term adaptations to caloric restriction (i.e., ↓IGF-I): but again, the use of a burst or plateau busting tactic for short term "diet sprints" at already low body fat seems the most logical approach for DNP & keto.

See Simkins S 1937 Dinitrophenol and desiccated thyroid in the treatment of obesity. JAMA 108:2110–2119:

> The extra energy of metabolism is derived mainly from fat and practically not at all from protein or carbohydrate. Consequently, dinitrophenol in therapeutic dosage produces no breakdown of significant amounts of body protein, even with patients on an inadequate protein intake. This is in marked contrast with the very consdierable increase in nitrogen excretion observed in patients undergoing treatment with thyroid. The fat is used completely and satisfactorily broken down, as no ketone bodies are found in urine. There is a no hyperlipidemia or constant change in the fixed and fatty acids of the blood.

pt. 1 of 2 pt. 2 of 2

Lethal doses for orally ingested 2,4-dinitrophenol in humans have been reported to be 14 to 43 mg/kg ... .

National Research Council. Drinking Water & Health, Volume 4. Washington, DC: National Academy Press, 1981., p. 237

Fatal dose in adults is about 1 to 3 g by mouth; 3 g has also proved fatal in divided doses over a period of 5 days. /Dinitrophenol/

Gosselin, R.E., R.P. Smith, H.C. Hodge. Clinical Toxicology of Commercial Products. 5th ed. Baltimore: Williams and Wilkins, 1984., p. III-157

Source: https://pubchem.ncbi.nlm.nih.gov/compound/2,4-dinitrophenol#section=Human-Toxicity-Values

The PK/PD of DNP in humans is considered unknown elimination half-life

Assumption:

• 36 hr elimination half-life (single dose)

: DNP accumulation given a single dose

Subject to multi-dose & dose-dependent pharmacokinetics:

> The elimination half-life (t1/2) of 2,4-DNP was (88.78±14.66) h in the routine HP group, while the t1/2 was only (54.58±12.92) h in the intensive HP group. The t1/2 of 2,4-DNP in the intensive HP group was apparently shorter than that in the routine HP group, with statistical significance (t=4.535, P=0.001). _[11]

Absorption, Distribution, Excretion (ADE): https://pubchem.ncbi.nlm.nih.gov/source/hsdb/529#section=Absorption-Distribution-and-Excretion-(Complete)

A single oral dose of from 3 to 5 mg per kilogram of body weight (3 - 5 mg/kg b.w.) produces an increase in RMR of from 20 to 30 percent within the first hour. This level is maintained for 24 hours and then falls gradually to normal on the third day. With daily administration there is a rapid rise of the RMR on the first day, a slight additional rise on subsequent days, and the maximum on the fourth day._[12]

After discontinuation, the RMR falls to normal within three to fourteen days._[12]

The response to a given dose may vary considerably from day to day._[12]

If the RMR rises less than 30 or 40 percent, there is little reaction except peripheral vasodilation, and a slight increase in respiratory minute-volume. A greater elevation produces uncomfortable heat and sweating._[12]

• 4 mg/kg results in ~40% increase to RMR

risk-reward

The use of 2,4-dinitrophenol ("DNP") may be rational dependent upon whether:

• ↑↑↑BMR/RMR dose-dependently (to the tune of 300 mg ⇒ ~33% ↑BMR [mean])
• Relative (i.e., muscle) protein sparing (preferentially oxidizes fat mass [FM])
• Improved blood lipids
• Improved glucose tolerance (but unclear and disparate effects on blood glucose levels or hyperglycemia)