San Antonio-based development company Wilmore Laboratories, LLC, is working to develop commercial uses for a novel, potentially highly effective formulation of resveratrol for nutraceutical delivery. A naturally occurring stilbene polyphenol found in grapes and other plant products and commonly associated with red wine, Resveratrol has attracted a great deal of research attention over the past decade for its potential to improve health and combat age-related diseases due to its physiological effects observed in animal studies. The compound has been shown to activate Sirtuin 1 (SIRT1) also known as NAD-dependent deacetylase sirtuin-1, a protein that in humans is encoded by the SIRT1 gene. With downstream effects as a vasodilator (via eNOS), an anti-inflammatory (via mTOR), and potential enhancement of athletic performance (via AMPK).
Wilmore Labs is currently focusing its attention on formulating Resveratrol for enhanced delivery to overcome the challenges shown in human studies as described in part by Wilmore’s science collaborator Dr. James Smoliga of High Point University in High Point, North Carolina. The company says it has developed a deep understanding interaction of Resveratrol with various molecules in compounding and in the human body, and from their scientific and product development experience, Wilmore Labs LLC is focused on development of a superior dosage form of Resveratrol to effectively and consistently enhance quality of life for the consumer market.
Reservatrol’s health benefits are still a subject of controversy in the research community. A study conducted at the University of Copenhagen and published online in July, 2013 in the Journal of Physiology gained international attention when it suggested that Resveratrol, a nutritional supplement widely regarded as agent responsible for health benefits of red wine, instead could actually cause harm by undoing the effects of exercise and increasing the risk of heart disease. The study was reported on in multiple press articles, but its validity is also being questioned.
On October 15, 2013, the journal published a letter from Dr. Smoliga and Wilmore Laboratories, LLC owner Otis L. Blanchard strongly challenging the study. (DOI: 10.1113/jphysiol.2013.262956 The Journal of Physiology
Volume 591, Issue 20, pages 5251–5252, October 2013) Their analysis suggests the conclusions drawn by the U. of Copenhagen researchers were unfounded, that no definitive conclusions were demonstrated, and that other studies have shown that Resveratrol may also have positive benefits for diabetes and heart disease. Mr. Blanchard notes, that “It’s a great example of how the scientific process continues after a study is published, and it’s a great thing when more researchers can openly discuss the implications a study.”
The original Copenhagen study declared itself to be the first to demonstrate negative effects of Resveratrol on [exercise] training-induced improvements in cardiovascular health parameters in humans, asserting that “Resveratrol supplementation was found to reduce the positive effect of exercise training on blood pressure, blood cholesterol and maximal oxygen uptake and did not affect the retardation of atherosclerosis.” The Smoliga/Blanchard letter strongly challenges these claims, noting that upon reviewing the study other researchers found that the differences were too small to support such a surprising conclusion, and that some of the study’s own data actually contradicts such negative findings, and that importantly, there were no post-training differences between groups for most of these, and it is not appropriate to interpret such results as statistical differences between groups. Their analysis showed that the total reduction was too slight to be mathematically certain, and that there was also no clinically meaningful difference between the subjects treated with Resveratrol and those treated with a placebo.
According to Dr. Smoliga in a Wilmore release, there were other issues with the study. There is a typical series of tests doctors use to diagnose and monitor atherosclerosis. This particular study did not use any of those procedures, and therefore would not be able to make any conclusions about atherosclerosis. The individuals studied did not have atherosclerosis in the first place, so it is impossible to reverse a medical condition that somebody does not have.”
Earlier in 2013, Dr. Smoliga authored an article in the journal Aging, explaining how research studies are often designed in such a way that they are unlikely to accurately analyze the effects of natural supplements in healthy people. Dr. Smoliga notes that “this really emphasizes the need for researchers to critically interpret studies and actively speak with the mainstream media, so that the public isn’t constantly confused by apparently conflicting results from scientific studies.”
A Mayo Clinic article entitled “Red wine and resveratrol: Good for your heart?” observes that “Resveratrol might be a key ingredient in red wine that helps prevent damage to blood vessels, reduces low-density lipoprotein (LDL) cholesterol (the “bad” cholesterol) and prevents blood clots.”
The article notes that most research on Resveratrol has been done on animals, not people, and research in mice given Resveratrol suggests that the antioxidant might also help protect them from obesity and diabetes, both of which are strong risk factors for heart disease. However, the article emphasizes that those findings were reported only in mice, not in people, and that in addition, to get the same dose of resveratrol used in the mice studies, a person would have to drink more than 1,000 liters of red wine every day. And while research in pigs has shown that Resveratrol may improve heart function and increase the body’s ability to use insulin, again, these benefits have not been tested in people.
On the other hand, the Mayo article observes that some research shows that resveratrol could be linked to a reduced risk of inflammation and blood clotting, both of which can lead to heart disease, but says that more research is needed before it’s known whether Resveratrol was the cause for the reduced risk, that one study showed that Resveratrol may actually reduce the positive effect of exercise on the heart in older men, and that it’s also important to know that resveratrol’s effects only last a short time after drinking red wine, so its effects may not last in the long term.
A new proof of concept study, funded by Wilmore Laboratories and published February 14 in the peer-reviewed journal PLOS One, titled “Development of a Lozenge for Oral Transmucosal Delivery of Trans-Resveratrol in Humans: Proof of Concept” (PLoS ONE 9(2): e90131. doi:10.1371/journal.pone.0090131) is coauthored by Dr. Smoliga and Mr. Blanchard, joined by Gregory Friesenhahn, and Martin A. Javors of the University of Texas Health Science Center at San Antonio Psychiatry department.
In the article abstract, the coauthors assert that Resveratrol provides multiple physiologic benefits that promote healthspan in various model species and clinical trials support continued exploration of resveratrol treatment in humans.
However, they note that there remains concern regarding low bioavailability and wide inter-individual differences in absorption and metabolism in humans, which suggests a great need to develop novel methods for resveratrol delivery. The researchers hypothesized that oral transmucosal delivery, using a lozenge composed of a resveratrol-excipient matrix, would allow resveratrol to be absorbed rapidly into the bloodstream, and pursued proof of concept through experiments on two human participants.
The solubility of trans-resveratrol (tRES) in water and 2.0 M solutions of dextrose, fructose, ribose, sucrose, and xylitol was determined, and the solubility of tRES in each of the solutions was compared to that in water, finding that tRES was significantly more soluble in a ribose solution than in four other solutions. Given the enhanced solubility of tRES in a ribose solution, a resveratrol-ribose matrix was developed into a lozenge suitable for human consumption. Lozenges were prepared, each containing 1465.5 mg tRES per 2000 mg of lozenge mass. Two healthy human participants consumed one of the prepared lozenges following an overnight fast, and venipuncture was performed immediately before and 15, 30, 45, and 60 minutes following lozenge administration.
The results observed suggest a resveratrol-ribose matrix lozenge can achieve greater Cmax and enter the bloodstream faster than previously reported dosage forms for gastrointestinal absorption, and while this study is limited by small sample size and only one method of resveratrol delivery, the coauthors contend that it does provide proof of concept to support further exploration of novel delivery methods for resveratrol administration.
The coauthors note that multiple published papers have described the results of tRES treatment in humans, and most research suggests that tRES is safe and could indeed improve cardiometabolic health, especially in individuals with chronic disease, although human bioavailability studies utilizing gastrointestinal absorption of resveratrol (i.e., ingestion of powders, capsules, caplets, etc.) have found wide inter-individual variability in tRES absorption, plasma concentration of free tRES, and resveratrol metabolite profile. They observe that It has long been known that tRES is extensively glycosolated and sulfalated during absorption in the small intestine, and that recent evidence suggests that gut microbiota may also play a role in metabolism of tRES before it reaches the bloodstream, contributing to inter-individual variability in bioavailability and metabolite profile. and while it is possible that unmodified tRES may demonstrate significant physiological effects once it reaches the plasma, the magnitude of these effects may be limited if absorption and metabolism limit plasma concentration and bioavailability. Consequently, oral transmucosal (OTM) delivery, which circumvents the gut and first-pass hepatic metabolism may increase the absorption of tRES and substantially reduce inter-individual variability in peak plasma concentration and metabolite profile to allow for greater clinical utility, and that an OTM formulation such as a lozenge could circumvent the physiological factors limiting absorption and bioavailability of tRES which normally occur following administration of a pill, food, or liquid.
The researchers note that Resveratrol has mixed potential as a candidate for OTM absorption, with favorable properties including relatively small molecular weight of tRES, and that tRES is uncharged at physiological pH which will allow passive diffusion across the buccal mucosa. A limiting factor for OTM formulations of tRES is dose size, which is restricted to approximately 100 150 mg for a lozenge, but this may be sufficient to achieve a physiologic effect if absorption is sufficiently high. Other OTM delivery forms, such as films and sprays are limited to a much smaller dose size.
The researchers hypothesized that incorporation of tRES into a simple lozenge would allow its adequate absorption through the oral mucosa of humans, and the first purpose of their study was to determine if the aqueous solubility of tRES would be affected by combining it with selected excipients to optimize the likelihood of significant OTM absorption. The second purpose of the study was to determine the maximal plasma concentration (Cmax) of tRES following administration of a tRES lozenge to healthy human volunteers. They report that results of these initial experiments provide proof-of-concept for the development of a tRES lozenge using appropriate excipients, which could lead to greater exploration of novel delivery methods for tRES administration and ultimately lead to new clinical applications.
Thus the first aim of the study was to determine if the aqueous solubility of tRES would be affected by dextrose, fructose, ribose, sucrose, or xylitol, and that although limited, the results provide some encouraging data to suggest that OTM administration may allow for more rapid absorption of tRES, which may have strong clinical applications. Likewise, a rapidly deliverable form of tRES may be beneficial in treating acute trauma to counter the development of oxidative stress and inflammation (e.g., concussion or spinal cord injury), which has also been supported through animal models.
In summary, the researchers conclude that results of this study demonstrate that ribose increases the aqueous solubility of tRES compared to its solubility in water alone. Further, OTM administration of a lozenge containing a mixture of ribose with resveratrol results in a very high peak plasma concentration compared to that reported from similar dosages of free resveratrol administered as a traditional oral supplement. Likewise, peak plasma concentration of free resveratrol was achieved approximately 15 minutes following lozenge administration, which is considerably quicker than the 60-120 minutes reported using traditional free resveratrol tablets.
They say that as human clinical trials investigate the benefits of tRES treatment on human health, it will become especially important to develop effective delivery methods which are cost effective, produce minimal side effects, and limit inter-individual bioavailability. Future research must be conducted to determine if the high plasma levels attained from the optimized resveratrol lozenge hold true across multiple individuals and whether the metabolite profile differs considerably from that obtained from an oral supplement.
Wilmore Laboratories, LLC