Elsevier

Phytomedicine

Volume 12, Issue 4, 20 April 2005, Pages 255-263
Phytomedicine

Formulation and in vitro/in vivo evaluation of combining DNA repair and immune enhancing nutritional supplements

https://doi.org/10.1016/j.phymed.2004.01.008Get rights and content

Abstract

Combining nutritional supplements to achieve synergistic benefit is a common practice in the nutraceutical industry. However, establishing added health benefit from a combination of natural ingredients is often assumed, untested and without regard to the principle of metabolic competition between the active components. Here, we report on the combination of a cat's claw water extract (C-Med-100, carboxy alkyl esters=active ingredients)+medicinal mushroom extracts (Cordyceps sinensis, Grifola blazei, Grifola frondosa, Trametes versicolor and Ganoderma lucidum, polysaccharides=active ingredients)+nicotinamide+zinc into a formulation designed to optimize different modes of immunostimulatory action, and yet that would avoid metabolic antioxidant competition yielding less than expected efficacious effects. Isobole curve analyses of these two active classes of ingredients determined by growth inhibition of HL-60 human leukemic cells in vitro confirmed they were indeed synergistic when in combination, and not metabolically competitive. Furthermore, an in vivo study showed significant health benefit for 14 subjects treated for 4 weeks with the unique C-Med-100/mushroom extract formulation in that they had reduced pain, reduced fatigue, weight loss and a reduced presence of DNA damage in peripheral blood assessed by (8-OH) guanine DNA adducts and elevation in serum protein thiols. Because this broad-based panel of clinical parameters indicating clinical efficacy has never been demonstrated before for either of the active ingredients evaluated alone in humans, these data were taken as strong evidence that the combination of C-Med-100+mushroom extracts+nicotinamide+zinc gave additive or synergistic effects to health benefit, and thus supported no efficacious limits from metabolic competition regarding this particular formulation.

Introduction

There are a wide variety of nutritional and dietary supplements that have been identified to possess clinically significant antiaging properties. Some of the most common preparations limit oxidative DNA damage by involving one or more metabolic pathways, and as a consequence increase cell responsiveness especially those regulating immune cellular reactions (Cross et al., 1987; Knight, 2000). For example, hot water cat's claw extracts such as C-Med-100 have been shown to not only reduce DNA damage by scavenging free radicals (Sandoval et al., 2000) but also by preventing their formation by inhibiting their generation via proinflammatory cytokine production through NF-kB (nuclear transcription factor beta) inhibition (Sandoval-Chancon et al., 1998). Such a dramatic reduction in DNA damage by inhibiting free radical production by this mechanism has been shown to simultaneously stimulate DNA repair and immune responsiveness (Sheng et al., 2000a, Sheng et al., 2000b, Sheng et al., 2001).

Parallel to this mechanism is the fact that DNA repair can also be stimulated by dietary nutritional factors such as zinc and niacin/nicotinamide. Low intracellular zinc resulting from nutritional deficiencies induce oxidative DNA damage, disrupt p53, NF-kB and AP-1 DNA binding, which in turn affect DNA repair (Ho and Ames, 2002). Likewise, niacin/nicotinamide are direct metabolic nutritional prescursors to the formation of cellular NAD (Jacob and Swendseid, 1996), which in turn is essential to energy production and it is also a co-substrate for the participation of poly (ADP-ribose) polymerase (PARP) in DNA repair (Rawling et al., 1994; Weitberg, 1989; Zhang et al., 1993). In other words, nutritional support for DNA repair enhancement is another pathway to resist DNA damage that is independent of either directly down-regulating free radical production by reducing NF-kB expression and thus the level of proinflammatory cytokines or by antioxidant free radical scavengers. In fact, nicotinamide and zinc supplements have been shown to enhance both DNA repair and immune responsiveness in vivo, thus confirming the health benefit of enhancing this mechanistic metabolic pathways (Sheng et al., 1998a, Sheng et al., 1998b).

Still another class of nutritional supplements to be considered for developing a broad spectrum mechanistic approach for an antiaging nutritional therapy are the natural occurring polysaccharides (i.e. beta glucans). Edible mushroom extracts especially those used in Chinese and Japanese natural medicines are a rich source of beta glucans. Thus, extracts of Cordyceps sinensis, Grifola blazei, G. frondosa, Trametes versicolor and Ganoderma lucidum all contain high amounts of polysaccharides which can directly stimulate immune reactions by primarily modulating immune responsive cytokines such as IL-1, IL-2, IL-6 and INF-gamma

(Borchers et al., 1999; Chen et al., 1997; Ebina and Fugimiya, 1998; Hsieh and Wu, 2001; Kiho et al., 1999; Mayell, 2001; Wang et al., 2002; Wasser and Weiss, 1999).

Among the many research and development questions that still remain regarding the health benefit of nutritional supplements is the scientific logic of combining them to increase their potency and biological effectiveness. As presented above, there are a lot of natural products that differ greatly in their chemistry, but that nonetheless can have similar health benefits such as lowering DNA damage and stimulating immune function. However, there are several metabolic and mechanistic pathways by which this could be accomplished. Here, it is important to remember the role human evolution has had to play in these matters. For example, humans have had to evolve by discovering ways to obtain nutritionous food from the environment. Among the earliest ways of deciding what to eat and not eat was by a “trial and error” approach; i.e. ingest it and if it had health or nutritionous benefits it became utilized with an increased frequency over time. Having this as a background to successfully populating highly diverse environments, it would have been important for our foraging ancestors to develop metabolic controls in order to prevent over exposure to biologically active dietary nutritional factors; i.e. nutritional and health benefiting substances that we need for sustaining life but not so much of them that they could limit our survival.

One mechanism consistent with our human evolution is that if our biological responses to ingestion of food sources were diversified so that several nutrients could be utilized to accomplish the same health benefit endpoint (e.g. immune function support), then there would be less dependence on any single nutritional factor limiting survival and the chances of over exposure and toxicity would be minimized. Metabolic diversity would then encourage metabolic competition. Otherwise it would be a serious limitation on survival to be exposed to several antioxidants in the food supply that all did the same thing thus becoming additive to the final end result such as scavengering to many free radicals. Thus having different metabolic pathways to limit free radical exposure (metabolic diveristy) through nutrient ingestion would be positive to survival. Moreover, if paralleled by having metabolic competition so that when more than one type of nutrient does the same thing such as scavenge free radicals, then the presence of one food source would prevent the uptake of another food source that did the same thing. There is already strong scientific evidence that metabolic competition of nutrients in fact exists. It has been shown that vitamins E, C and carotenoids are different chemical versions of naturally occurring free radical scavengers that can strongly regulate each other's uptake (Baker et al., 1996; Niki et al., 1995).

Taken together there seems to be credible evidence that combinig nutrients having similar modes of action will not necessarily produce synergized blends of nutrients designed to improve the effectiveness of broad spectrum antiaging therapies, basically because of metabolic competition. However, taking advantage of the metabolic diversity of combining several nutrients having varied metabolic pathways to mediate desired health benefits by stimulating the natural processes of DNA repair and immune functions, seems logical to help nutritionally protect individuals against a spectrum of diseases associated with aging such as cancer, inflammation, diabetes, autoimunne and cardiovascular disorders. Here, we report on combining a metabolically diverse nutrient spectrum containing nicotinamide, zinc, C-Med-100 (cat's claw water extract), and five mushroom extracts (C. sinensis, G. blazei, G. frondosa, T. versicolor and G. lucidum) into a single orally administered capsule for evaluation of clinical outcome using several subjective questionaire and biochemical endpoints.

Section snippets

Nutraceutical product

The trade name of the nutraceutical product designed to combine metabolically diverse natural ingredients that can enhance DNA repair and immune function was manufactured by Phoenix Laboratories, Inc (Hicksville, NY) and originally called Agerasers, but recently altered and sold as Xenodrine 40+ after addition of norambrolide (Cytodyne). It contained the following active ingredients per capsule (total=560 mg): C-Med-100=200 mg, nicotinamide=100 mg, Opti-zinc (20%-international)=10 mg, and KMA

In vitro synergy of C-Med-100

Before proceeding with the in vivo evaluation of the C-Med-100/mushroom extract formulation, in vitro analyses that the combination of C-Med-100 with mushroom extracts would provide data that the active ingredients combined in the C-Med-100/mushroom extract formulation were in fact synergistic to each other. Table 2 presents the data for such an anlaysis. It seems apparent that C-Med-100 formula (i.e. Nicomed) combined with KMA complex gave lower than expected IC50 values, clearly establishing

Discussion

The primary aim of this study was to demonstrate the added benefit of developing a nutritional supplement designed to take advantage of metabolic diverse pathways that can combat the hazardous health consequences of oxidative stress by different mechanisms. C-Med-100/mushroom extract formula is such a product whereby the health benefits of DNA repair and immune enhancements which are in turn modulated by antioxidizaton can be further enhanced by the combination of ingredients that represent

Acknowledgements

The authors are grateful to the staff at the Department of Cell and Molecular Biology, Lund University, Lund, Sweden and to ImmunoSciences, Inc., Beverly Hills, CA as well as to Dr. Margaretha Lund-Pero for their technical support. We are also thankful to Phoenix Labs, Inc and Campamed, LLC for their financial support.

References (32)

  • A.B. Weitberg

    Effect of nicotinic acid supplementation in vivo on oxygen radical-induced genetic damage in human lymphocytes

    Mut. Res.

    (1989)
  • E.M. Williamson

    Synergy and other interactoins in phytomedicines

    Phytomedicine

    (2001)
  • J.Z. Zhang et al.

    Poly(ADP ribose) polymerase activity and DNA strand breaks are affected in tissues of niacin-deficient rats

    J. Nutr.

    (1993)
  • H. Baker et al.

    Human plasma patterns during 14 days ingestion of vitamin E, beta-carotene, ascorbic acid, and their various combinations

    J. Am. Coll. Nutr.

    (1996)
  • M. Berenbaum

    What is synergy?

    Pharmacol. Rev.

    (1989)
  • A.T. Borchers et al.

    Mushrooms, tumors, and immunity

    Proc. Soc. Biol. Med.

    (1999)
  • Cited by (0)

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