Could Compound 14 mimic exercise?
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Could ‘Compound 14’ be the diet pill that mimics exercise?

Compound 14 shows potential as a weight-loss aid with studies supporting efficacy, but it could take a long time to be marketed as a supplement.

Introduction

A regular exercise routine is the most reliable way for anybody to lose weight. Not only does exercising directly increase the body’s metabolic rate of carbohydrate, protein and fat, it can lead to increased energy consumption afterwards and helps build muscle which consumes still more calories. Until recently, no chemicals have been synthesised that activated exactly the same pathways which lead to increased metabolic rate during exercise, though supplements with effects similar to adrenaline, the body’s exercise-preparation hormone, have existed for years. ‘Compound 14’ has been described as the first chemical to induce the same metabolism-boosting effects as intense exercise.

What is Compound 14?

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Compound 14 is a chemical recently developed at the University of Southampton, UK, which studies on mice have demonstrated is effective for treating obesity. The chemical inhibits enzymes inside cells that are related to metabolism, causing a build-up of chemicals that the cell interprets as a lack of food; the cells subsequently start absorbing glucose from the blood and boosting their metabolic rate. Specifically, compound 14 is based on a cyclic protein, and was actually developed in previous projects.

In cells, an enzyme named AMP-activated protein kinase (AMPK) is responsible for governing the metabolism enhancing effects observed with exercise. AMPK sets off a ‘chain reaction’ of processes inside the cell, leading to increased uptake of glucose from the blood (by increased insulin production), increased usage of fatty acids and stored carbohydrates for fuel, inhibition of lipogenesis and triglyceride synthesis (reduced body-fat being created), and prevention of cholesterol synthesis. AMPK has been linked to exercise-induced weight loss, with recent studies even suggesting AMPK could be responsible for increasing blood supply to the muscles. AMPK is not normally activated in cells, but becomes active when it interacts with ZMP, another cellular chemical. ZMP has been used therapeutically in the past via injection into cells in a form that can get through the cell membrane, AICAR; though this activates AMPK, the effects are short lived because AICAR breaks down quickly, and there is a dangerous increase in blood lactic and uric acid. ZMP is naturally produced during cell metabolic processes, but is usually kept at low levels by the enzyme ATIC. ATIC needs to homodimerise (bind to another ATIC molecule), to form the enzyme AICART, before it can become an active enzyme and break down ZMP; the researchers hypothesised that inhibiting the homodimerisation of ATIC could be an effective way of activating AMPK. If AICART is not formed, then ZMP levels increase in the cell, and AMPK is activated to the extent that significant metabolic changes are observed. Compound 14 was discovered to be able to inhibit the formation of AICART, and so the researchers conducted an experiment into its weight-loss properties in mice.

How is Compound 14 different to other diet pills and ingredients?

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As described, compound 14 is not a diet supplement, or even an ingredient at this point in time; it is a synthetic chemical which has been thus far only administered to cells and mice in an external environment and via peritoneal (body) injections respectively. Compound 14 is relatively unusual amongst chemicals that are known for weight loss properties, in that its pathway is both entirely understood and it is proven to be effective for weight loss, with (at this point) no conflicting studies suggesting otherwise. Additionally, being synthetic, compound 14 is not available through any source in nature, whereas most diet supplements tend to use natural ingredients because they are easier to source; it is likely that a compound 14 supplement would be very expensive as synthesising the chemical would be more complicated than isolating it from a natural source. The main difference is that compound 14 was designed specifically to induce weight loss, whereas most ingredients serve a purpose in their natural source and happen to induce weight loss in humans.

Clinical studies of Compound 14

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Compound 14 has had two main studies on its effects – the study describing the origins and structure of compound 14 and its potential for use in cancer therapy, and the study detailing its efficacy for weight loss, which involves both mammalian and cell culture trials. The first study, from 2012, described the pathway by which ATIC can be prevented from homodimerising, thus preventing the creation of AICART and leading to increased activity of AMPK ; the researchers explain that ATIC is formed from two sections, with amino acid chains that can lock over one another, forming a bond that leads to AICART. The ATIC molecules actually have active enzyme areas attached to them which swap amino acid structures, leading to a reaction that forms the bond; this is where AICART’s active site is found, which is used to break down ZMP. The researchers had previously reviewed a library of millions of chemicals with the right structure to be a potential inhibitor of ATIC homodimerisation; they chose compound 14 as it was the most potent. The researchers screened its structure and found that despite having multiple regions which were suitable for inhibiting ATIC homodimerisation, it could be improved by changing the amino acids to some not normally found in nature; this resulted in a 120-fold improvement in activity. Having improved the molecule, the researchers then performed a trial in breast cancer cells. Breast cancer is in part caused or enhanced by the inhibition of certain enzymes in cells, which is believed to be mediated by AMPK. In a study of a cell culture of breast cancer cells, solutions of compound 14 were added; a dose-dependent reduction in cell number was observed over a 72 hour period. The researchers concluded that compound 14 could be a potent inhibitor of ATIC homodimerisation.

The second study determined that compound 14 could have exercise-simulating effects. Similar the first study, the researchers exposed MCF-7 cells (breast cancer cells) to a solution of compound 14, and observed the cellular increase in ZMP over the next few hours; a threefold increase was observed over 3 hours. A fluorescent probe was added that tested for AMPK activation; a 1.18 fold increase in AMPK activity was observed over the same time period. This was almost identical to the change observed when a direct activator of AMPK was tested. However, the researchers noted that there were two possible means of breaking down ZMP, and tested compound 14 in conditions where AICART was the main enzyme, and where another enzyme was the main enzyme, respectively. Compound 14 was effective in the first case, but not the second; this means compound 14 will not act on all cells of the body, as some rely on the second mechanism. The researchers then tested compound 14 for ability to kill cancer cells, with two cancerous cell strains being exposed to the chemical. With a relatively high dose, around 40% of cancer cells died from exposure. However, further tests revealed that this was due to cytostatic, rather than cytotoxic, effects – i.e. the cells were prevented from multiplying, rather than killed directly, which makes compound 14 safer to use for weight loss.

Compound 14 was then tested for effects on oxygen consumption and lactic acid production. An increase of 1.4 fold in oxygen consumption in breast cancer cells, and a 1.25 decrease in lactate levels, were observed; lactate is the conjugate base of lactic acid, meaning lactic acid levels were higher in the cells (which is associated with exercising). This indicated a greater level of respiration in the cells, meaning greater energy usage. Finally, the main study of interest saw 15-week old mice fed either standard food or a high-fat diet, until the high-fat mice were obese. Injection into both groups of mice revealed that the blood glucose levels of mice of normal bodyweight were reduced, on average by 21%; however, the blood glucose levels of the high-fat mice were shown to be both significantly higher than those of the normal mice before treatment, and reduced by 42% on average following compound 14 injection. However, continuous treatment with compound 14 over 7 days revealed negligible effects on the blood glucose levels of normal mice, but significant effects on those of obese mice, with a huge decrease in resting blood glucose levels. Finally, the mice were treated daily for seven days, and the results showed an increase in weight of the normal-diet group of mice with the control, and a slight decrease (just over 1%) in weight for compound 14. However, the high-fat fed group, with an average weight of 35.8 grams, saw an average decrease in bodyweight of 1.62 grams, a loss of just under 5%. This was despite continued consumption of a high-fat diet.

In the discussion, the researchers commented that the pathways ZMP operates by are not fully understood and require more research, and that other drugs which activate AMPK do so through different pathways; this means that more drugs might be synthesisable that target AMPK, which could give patients more choice and make treatment for accessible. The researchers also commented that the high-fat mice which lost a considerable quantity of weight did not exhibit any negative side effects or loss of appetite. AMPK, is able to uncouple proteins in adipose tissue, making the internal fatty acids more available for digestion- the researchers clarify, however, that body fat percentage was not measured, and that further studies are needed to test for a loss of actual adipose tissue. This implies that AMPK could cause reductions in fat, carbohydrate, protein and even bone density levels. However, since similar weight loss was not observed in the normal group, this was decided to be unlikely.

Safety considerations and side effects of Compound 14

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The pioneering studies on compound 14 have not revealed any undesirable effects; the mice used in the study did not exhibit any malaise and no casualties were incurred. However, thus far the studies have focused on the effects of compound 14 in the short term (for the weight loss and glucose control studies) and in cancerous cells (for the anti-tumour study) respectively. Long-term use of compound 14 could lead to issues, and further research will need to be conducted to confirm this; this is because the researchers are not sure exactly how the pathways leading to AMPK activation work, and note that ZMP can be created through multiple pathways, meaning compound 14 could be having unknown effects in the cells which could make them unsuitable for certain individuals. The effects on breast cancer cells are unlikely to be indicative of destructive or damaging potential of compound 14, for two reasons: firstly, the cancer cells were not killed, but simply stopped multiplying incorrectly; and secondly, the cancer cells developed due to failure of AMPK activation, which compound 14 corrected, and normal cells have functional AMPK activation. All drugs require extensive testing, particularly human trials, before they can be released to the public; but at this stage it appears side effects of compound 14 are unlikely.

What’s the next step?

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Unfortunately, the procedure in getting a drug from research to market is extensive. Having had clinical studies conducted, the sponsors of the research – the University of Southampton – will have to present their research to the ‘Medicines and Healthcare products regulatory agency’, who will try and get legislation pushed through in government for the drug to be marketable. However, since clinical studies have already taken place and the drug has proven effective, it is likely the drug will be made available without serious resistance. Whether it will be available over-the-counter remains to be seen.

Conclusion

Compound 14 has so far been demonstrated to be an effective anti-diabetic and weight loss aid, which also possesses anti-cancer potential. It has also thus far proven safe and the proposed pathway does not include any potentially dangerous activity, though long-term studies will be needed to assess both efficacy and side effects. Since compound 14 is a research drug and has only recently been explored for weight loss, it is unlikely to reach the pharmaceutical market in a short period of time, and may only be available for obese patients due to its higher efficacy in individuals with higher body-fat percentage.

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