Marketing Claims and Research Strategies
in the Nutraceutical Industry
Gitte Jensen, PhD and Aaron Hart, NIS Labs
Research in the natural products industry is on a great track, yet must contend with challenges that are not often seen in the pharmaceutical industry. These challenges are often due to the heterogeneous nature of nutraceutical products. This complexity has technical and marketing advantages but requires diligence and creativity in research design and implementation.
The need for continued scientific research on natural products remains critical. Standard pharmaceutical methods are not always applicable for assessment of natural products. DSHEA places significant burden on the dietary supplement industry, limiting claims to structure and function claims. Given the increased demand for proof of efficacy, it is often difficult to demonstrate efficacy without exceeding the structure and function claims and wander toward health claims.
In order to deliver reliable results that support claims compliant with DSHEA, dietary supplements must be studied using scientific methods that are well suited for each dietary supplement. Research-based proof of marketing claims in the dietary supplement industry is now an integral part of marketing standardized products with demonstrated efficacy.
The pharmaceutical model of scientific research that is often held as a reference does not always apply to the dietary supplement industry because the dietary supplement and pharmaceutical industries aim at different claims, have different economics and require different levels of proof before going to market. Research in the dietary supplement industry must be stringent and solid, yet creative and adapted to working with the complexity of natural products, and deliver results that will support structure and function claims as well as satisfy the increasing demand for proof of efficacy.
Natural Immune Systems (NIS) Labs Inc. focuses on in vitro studies in cell-based test models that produce information related to mode of action that help formulate structure and function claims compliant with DSHEA.
Product Preparation for Cell Culture
A safe working range must be established in liquid formulation appropriate for cell work before products may be incorporated into cell culture. It is crucial to ensure that the product is tested at a concentration that is high enough to show an effect, but low enough to avoid problems with culture pH, tonicity and general cytotoxicity. Interference from high salt concentrations or the formation of precipitates must be assessed and addressed.
Safety Issues
The first step after developing appropriate test conditions is often to determine whether or not the product will initiate a mitogenic response in human cells. A mitogen is a compound that triggers a strong and non-discriminative immune reaction. This test is particularly relevant to include in applications for FDA approval of a new dietary ingredient.
Customized Research Protocols
Designed to Elucidate the Mode of Action
- Anti-Inflammatory Action - Research Method: COX-2
The current focus on COX-2 inhibitors in both the pharmaceutical and natural products industries suggests the need for thorough planning and investigation into the specific activity of new anti-inflammatory products. Some natural products, such as Curcumin, inhibit both gene expression and enzymatic activity of COX-2. Resveratrol inhibits COX-2 induction by some inflammatory signals but not others, suggesting a selective, possibly tissue-specific, anti-inflammatory effect instead of a global inhibition.
Methods for Identification of Mechanism of Action of COX-2 Inhibitors: Overview and Approaches:
Cell-based assays can demonstrate anti-inflammatory effects by looking at COX-2 gene expression levels. Compounds that block this type of activity include xanthones such as are found in mangosteen fruit. Methods directed at this mechanism include detection of intracellular COX-2 by monoclonal antibodies and flow cytometry, or detection of mRNA for COX-2.
A cell-free assay using human recombinant COX-2 enzyme and appropriate substrates for prostaglandin production can be used to specifically address whether an extract contains compounds that interfere with COX-2 enzymatic function. Examples of compounds that block enzyme function directly include over the counter drugs like acetaminophen, and ibuprofen. This assay does not address whether the extract inhibits COX-2 gene expression.
Prostaglandin levels can be measured by ELISA on plasma/supernatant from whole blood cultures. This testing does not on its own provide information on whether inhibition of prostaglandin production happens at the level of gene expression or enzymatic activity. However, by combining this test with the two tests above, conclusions can be made as to mechanism of action of a COX-2 inhibitor.
- Immune Activation - Research Method: NK Cell Activation
Activation of immune function is a common target for nutraceutical claims. The interplay between the many subsets of cells and tissues involved in host mediated response to various diseases is a complicated puzzle. Immunomodulatory effects can be at least partially addressed by looking at specific changes in activation markers in different cell types (NK cells, B cells, T cells, etc).
An example of testing mechanism of action involves NK cell activation, leading to structure/function claims of immune enhancement. Using purified NK cells, it can be tested whether the effect is directly on NK cells, and a search for specific receptor involvement can be undertaken.
Pharmaceutical drugs are often tested for whether they act as agonist or antagonist in a certain physiological system. The heterogeneity of many natural products leads to more complex action, such as modulating signaling pathways at multiple locations, resulting in an altered response to known stimuli.
- Cell Signaling - Research Method: Cell Signaling
Understanding the effects of a product on the interplay between cell signaling pathways is one of the most complex and concrete type of information that can be presented about a nutraceutical product. This type of research describes the “how” of observed in vitro effects through the identification of physical mechanisms for immunomodulatory function. Aside from the inherent complexity of even the simplest pathways, it is important to consider cell type as well as interactions between cell types when determining appropriate assay types and conditions.
Methods for Studying Signaling Pathways by Natural Products and Compounds
Receptor/Ligand studies are used to identify direct binding of compounds form a natural product to cell surface molecules.
Cell-based assays are used to assess altered expression of receptors for chemical attractants dictating immune cell trafficking.
Intracellular and intercellular signaling molecules (such as calcium) are measured.
The degree of phosphorylation on signaling compounds can be quantified.
Messenger RNA expression can be qualitatively and semi-quantitatively determined through the use of micro array technology.
Products of these pathways can often be consistently detected in cell lysates and quantified by ELISA.
An inflammatory mediator such as TNF-a is bound to its receptor which triggers a massive signaling complex which ultimately results in the degradation of IKK and the recruitment of NF-kB to the nucleus of the cell where it promotes the expression of a specific set of genes in a pre programmed response.