Biomedica Laboratories, Recovery

Clinical Research

Safety Data

Recovery has significant benefits with very low risk. All Recovery ingredients are naturally-occuring and non-toxic.

Nutricol constituents have been clinically observed to possess anticarcinogenic properties in the liver, lung, breast, pancreas, bladder, prostate, skin and most of the gastrointestinal system (Fujiki. (1999) J. Cancer Res Clin Oncol.125:589-97).

Effects on Liver Function

Due to anti-catabolic and anti-oxidative actions, Recovery may aid in the proper elimination and metabolism of drugs and other toxins by supporting 4 Phase II liver pathways (glutathione conjugation, taurine conjugation, methylation, and sulfation).

Anti-inflammatory (NSAIDs/cox-2 inhibitors)

Most conventional NSAIDs interfere with cyclo-oxygenase and prostaglandins. Cell damage still continues because:

• Oxidation of membranes remains unblocked
• With standard NSAIDs, the production of PG1 and PG3, normally involved in repair, are also blocked

Recovery benefits alone or combined with NSAIDS include:

Inhibiting the inflammatory cascade or “domino effect” by increasing a cell’s ability to neutralize lysosomal enzymes and ROS released from neighboring damaged cells - reducing trauma.

Increasing delivery of certain hormones, neurochemicals and nutrients into the cell and enhancing waste transport out of the cell - improving cell communication.

Studies have shown that the addition of ingredients in Recovery with Sulindac (NSAID) results in a synergistic effect on prevention of colon cancer in rats and a reduction in GI side-effects that accompany Sulindac usage (Ohishi et al. Cancer Lett 2002, 177(1): 49-56)

Corticosteroids

Corticosteroids mimic cortisol, which reduces infl ammation; however, corticosteroids inhibit immune response and ability to repair, predisposing individuals to risk of infection and accelerated rate of tissue breakdown.

Excessive levels of nitric oxide synthase (NOS), an enzyme that produces nitric oxide, are involved in the initiation and progression of cancer and infl ammation. Studies have shown higher levels of nitric oxide in various infl ammatory bowel diseases, and that corticosteroids have no effect on reducing NOS. (N Leonard, et. al. J. Clin. Pathology: 1998, 51: (10) 750-753)

Recovery may compliment corticosteroids as it can normalize levels of NOS (Yu-Li Lin et.al. Molecular Parm: 1997 (52):465-472).

Acetaminophen

Recovery ingredients reduce acetaminophen-induced kidney and liver toxicity (Res Commun Mol Pathol Pharmacol 2000; 107(1-2):137-66), (Ray S.D., Arch Biochem Biophys 1999 Sep 1; 369(1):42-58).

Many cases have demonstrated Recovery may be superior to acetaminophen for chronic pain relief.

Recovery decreases the need for acetaminophen

Antibiotics

2 studies report anti-bacterial action was enhanced when Recovery ingredients were combined with ampicillin/ sulbactam, benzylpenicillin, oxacillin, methacillin, cephalexin (Journal of Antimicrobial Chemotherapy, 2001, (48), 361-364), (Antimicrobial Agents and Chemotherapy, 2001, 45, (6), 1737-1742).

Tamoxifen

2 studies report an enhanced anti-cancer effect when Recovery ingredients were combined with Tamoxifen (Suganuma M., Biofactors 2000:13(1-4): 67-72), (Fujiki H., 1999 Society for Experimental Biology and Medicine, Vol. 220, 225-228).

Anti-coagulants

Over the last 5 years, Biomedica has made observations with several patients on warfarin and Recovery. There were no changes in prothrombin time reported, nor any signs of increased bleeding. In vitro studies show no effect on thromboplastin times or prothrombin times. Recovery may have anti-platelet activity related to normalizing excessive platelet adhesiveness. (Kang WS., Thromb Res 1999 Nov 1; 96(3):229-37)

Amiodarone, Doxorubicin, Idarubicin, 4-HC

The ingredients in Recovery reduce organ and serum toxicity induced by these drugs (Bagchi D., Drugs Exp Clin Res 2001; 27(1): 3-15), (Res Commun Mol Pathol Pharmacol 2000; 107(1-2): 137-66)

Bioflavonoids & Osteoarthritis

Singh R, Ahmed S, Malemud CJ, Goldberg VM, Haqqi TM, J Orthop Res 2003 Jan;21(1):102-9 Epigallocatechin-3-gallate selectively inhibits interleukin-1beta-induced activation of mitogen activated protein kinase subgroup c-Jun N-terminal kinase in human osteoarthritis chondrocytes.

Activation of mitogen activated protein kinases (MAPK) is a critical event in proinflammatory cytokine-induced signalling cascade in synoviocytes and chondrocytes that lead to the production of several mediators of cartilage damage in an arthritic joint. Green tea (Camellia sinensis) is a widely consumed beverage and we earlier showed that polyphenols present in green tea (GTP) inhibit the development of infl ammation and cartilage damage in an animal model of arthritis. In this study we evaluated the role of epigallocatechin-3-gallate (EGCG), a green tea polyphenol which mimics its antiinflammatory effects, in modulating the IL-1beta-induced activation of MAPK’s in human chondrocytes. We discovered that EGCG inhibited the IL-1beta-induced phosphorylation of c-Jun N-terminal kinase (JNK) isoforms, accumulation of phospho-c-Jun and DNA binding activity of AP-1 in osteoarthritis (OA) chondrocytes. Also IL-1beta, but not EGCG, induced the expression of JNK p46 without modulating the expression of JNK p54 in OA chondrocytes. In immunocomplex kinase assays, EGCG completely blocked the substrate phosphorylating activity of JNK but not of p38-MAPK. EGCG had no inhibitory effect on the activation of extracellular signal-regulated kinase p44/p42 (ERKp44/p42) or p38-MAPK in OA chondrocytes. EGCG or IL-1beta did not alter the total non-phosphorylated levels of either p38-MAPK or ERKp44/p42 in OA chondrocytes. Conclusion: These are novel findings and indicate that EGCG may be of potential benefit in inhibiting IL-1beta-induced catabolic effects in OA chondrocytes that are dependent on JNK activity.

Ahmed S, Rahman A, Hasnain A, Lalonde M, Goldberg VM, Haqqi TM, Free Radic Biol Med 2002 Oct 15;33 (8):1097-105 Green tea polyphenol epigallocatechin-3-gallate inhibits the IL-1 beta-induced activity and expression of cyclooxygenase-2 and nitric oxide synthase-2 in human chondrocytes.

We have previously shown that green tea polyphenols inhibit the onset and severity of collagen II-induced arthritis in mice. In the present study, we report the pharmacological effects of green tea polyphenol epigallocatechin-3-gallate (EGCG), on interleukin-1 beta (IL-1 beta)-induced expression and activity of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in human chondrocytes derived from osteoarthritis (OA) cartilage. Stimulation of human chondrocytes with IL-1 beta (5 ng/ml) for 24 h resulted in signifi cantly enhanced production of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) when compared to untreated controls (p <.001). Pretreament of human chondrocytes with EGCG showed a dose-dependent inhibition in the production of NO and PGE(2) by 48% and 24%, respectively, and correlated with the inhibition of iNOS and COX-2 activities (p <.005). In addition, IL-1 beta-induced expression of iNOS and COX-2 was also markedly inhibited in human chondrocytes pretreated with EGCG (p <.001). Parallel to these fi ndings, EGCG also inhibited the IL-1 beta-induced LDH release in chondrocytes cultures. Conclusion: Overall, the study suggests that EGCG affords protection against IL-1 beta-induced production of catabolic mediators NO and PGE (2) in human chondrocytes by regulating the expression and catalytic activity of their respective enzymes. Furthermore, our results also indicate that ECGC may be of potential therapeutic value for inhibiting cartilage resorption in arthritic joints.

Singh R, Ahmed S, Islam N, Goldberg VM, Haqqi TM, Arthritis Rheum 2002 Aug; 46 (8): 2079-86 Epigallocatechin gallate inhibits interleukin-1beta-induced expression of nitric oxide synthase and production of nitric oxide in human chondrocytes: suppression of nuclear factor kappaB activation by degradation of the inhibitor of nuclear factor kappaB.

Human chondrocytes were derived from OA cartilage and were treated with EGCG (100 microM) and IL-1beta (2 ng/ml) for different periods, and inducible nitric oxide synthase (iNOS) messenger RNA and protein expression was determined by real-time quantitative reverse transcriptase-polymerase chain reaction and Western blotting, respectively. Production of NO was determined as nitrite in culture supernatant. Activation and translocation of nuclear factor kappaB (NF-kappaB), levels of inhibitor of nuclear factor kappaB (IkappaB), and NF-kappaB DNA binding activity were determined by Western blotting and a highly sensitive and specifi c enzyme-linked immunosorbent assay. Activity of IkappaB kinase was determined using in vitro kinase assay. Human chondrocytes cotreated with EGCG produced signifi cantly less NO compared with chondrocytes stimulated with IL-1beta alone (P < 0.005). The inhibition of NO production correlated with the suppression of induction and expression of NF-kappaB-dependent gene iNOS. EGCG inhibited the activation and translocation of NF-kappaB to the nucleus by suppressing the degradation of its inhibitory protein IkappaBalpha in the cytoplasm. Conclusion: Our results indicate that EGCG inhibits the IL-1beta-induced production of NO in human chondrocytes by interfering with the activation of NF-kappaB through a novel mechanism. Our data further suggest that EGCG may be a therapeutically effective inhibitor of IL-1beta-induced infl ammatory effects that are dependent on NFkappaB activation in human OA chondrocytes.

Takita H, Kikuchi M, Sato Y, Kuboki Y, Connect Tissue Res 2002;43(2-3):520-3 Inhibition of BMP-induced ectopic bone formation by an antiangiogenic agent (epigallocatechin gallate)

Epigallocatechin gallate (EGCG), which is one of the components of green tea, was recently shown to inhibit endothelial cell growth in vitro and angiogenesis in vivo [5]. We have previously shown that bone and cartilage formation by bone morphogenetic protein (BMP) is highly dependent on the geometry of the carrier (vasculatureinducing or -inhibiting geometry [2]. To verify the function of angiogenesis in the BMP induction system, we examine in this article whether inhibition of angiogenesis enhances chondrogenesis and suppresses osteogenesis. Fibrous glass membrane used as a BMP carrier was mixed with 1.2 micrograms rhBMP-2 and 1-10 micrograms of EGCG and was implanted into rats subcutaneously. As the dose of EGCG increased, alkaline phosphatase activity and calcium content were decreased, whereas the type II collagen content was increased. Conclusion: The results clearly indicated that inhibition of vascularization enhanced chondrogenesis and suppressed osteogenesis. Chen PC, Wheeler DS, Malhotra V, Odoms K, Denenberg AG, Wong HR, Infl ammation 2002 Oct; 26 (5):233-41 A green tea-derived polyphenol, epigallocatechin-3- gallate, inhibits IkappaB kinase activation and IL-8 gene expression in respiratory epithelium.

Interleukin-8 (IL-8) is a principle neutrophil chemoattractant and activator in humans. There is interest in developing novel pharmacological inhibitors of IL-8 gene expression as a means for modulating infl ammation in disease states such as acute lung injury. Herein we determined the effects of epigallocatechin-3-gallate (EGCG), a green teaderived polyphenol, on tumor necrosis factor-alpha (TNF-alpha)-mediated expression of the IL-8 gene in A549 cells. EGCG inhibited TNF-alpha-mediated IL-8 gene expression in a dose response manner, as measured by ELISA and Northern blot analysis. This effect appears to primarily involve inhibition of IL-8 transcription because EGCG inhibited TNFalpha- mediated activation of the IL-8 promoter in cells transiently transfected with an IL-8 promoter-luciferase reporter plasmid. In addition, EGCG inhibited TNF-alphamediated activation of IkappaB kinase and subsequent activation of the IkappaB alpha/NF-kappaB pathway. Conclusion: We conclude that EGCG is a potent inhibitor of IL-8 gene expression in vitro. The proximal mechanism of this effect involves, in part, inhibition of IkappaB kinase activation.

Adcocks C, Collin P, Buttle DJ, J Nutr 2002 Mar; 132 (3):341-6 Catechins from green tea (Camellia sinensis) inhibit bovine and human cartilage proteoglycan and type II
collagen degradation in vitro.

Polyphenolic compounds from green tea have been shown to reduce inflammation in a murine model of infl ammatory arthritis, but no studies have been undertaken to investigate whether these compounds are protective to joint tissues. We therefore investigated the effects of catechins found in green tea on cartilage extracellular matrix components using in vitro model systems. Bovine nasal and metacarpophalangeal cartilage as well as human nondiseased, osteoarthritic and rheumatoid cartilage were cultured with and without reagents known to accelerate cartilage matrix breakdown. Individual catechins were added to the cultures and the amount of released proteoglycan and type II collagen was measured by metachromatic assay and inhibition ELISA, respectively. Possible nonspecific or toxic effects of the catechins were assessed by lactate output and proteoglycan synthesis. Catechins, particularly those containing a gallate ester, were effective at micromolar concentrations at inhibiting proteoglycan and type II collagen breakdown. No toxic effects of the catechins were evident. Conclusion: We conclude that some green tea catechins are chondroprotective and that consumption of green tea may be prophylactic for arthritis and may benefi t the arthritis patient by reducing inflammation and slowing cartilage breakdown. Further studies will be required to determine whether these compounds access the joint space in suffi cient concentration and in a form capable of providing efficacy in vivo.