A MEDICAL FOOD
Since its discovery in 1999, intensive research has been performed on ghrelin in humans without any evidence of overt toxicity. To the contrary, ghrelin has been shown to be effective in opposing or reducing various side effects with diverse causes ranging from drug induced cardiotoxicity to ethanol and acetamenophen tissue damage (1-3). Clinical studies designed to identify possible toxic effects of ghrelin have failed to do so. For example, in a double-blind, randomized, placebo-controlled trial in which young, healthy male subjects receiving low, moderate and high doses of ghrelin intravenously, the number of subjects that experienced adverse effects did not significantly differ among the groups, that the results suggest ghrelin is safe for human use and that it may be useful for the treatment of disorders ranging from GH secretion to appetite control (4). In a more recent clinical trial (5) designed to assess the beneficial effects of ghrelin in cancer patients, drug-related adverse events (assessed by NCI-CTC-toxicity criteria and cardiac examination) did not differ between ghrelin and placebo leading the authors to report that “ghrelin is well tolerated and safe, even in patients with advanced cancer”.
Similarly, peptidyl analogs of ghrelin such as GHRP-2 that are GHS receptor agonists are essentially non-toxic. When the peptide was administered in graded doses to children, it was found to be well tolerated, lacking in side effects and toxicities after 8 consecutive months of treatment (6). This lack of toxicity has been reported for other GHS agonists and seems to be characteristic of this family of molecules. For example, the GHS receptor agonist and ghrelin analog capomorelin (7) was well tolerated and lacked toxicity in a total of 18 clinical studies. Furthermore, when subjected to a battery of toxicity tests, GHRP-2 was found to be benign in all and comparable to placebo control measures. For example, when tested for acute oral toxicity (LD50), the peptide was found to be safe at 2000 mg/kg when administered to Wistar rats as two divided dose within a one hour interval. Hence it was declared to be in GHS-5/unclassified category. Furthermore, after repeated oral administration of GHRP-2 for 90 days, there were no adverse effects observed on any parameters of study and a No Observed Adverse Effect Level (NOAEL) for the test article was found to be 50 mg/kg. Using the bacterial reverse mutation assay (AMES test) GHRP 2 was non mutagenic up to dose of 2 mg/ml. The results of mammalian bone marrow micronucleus study and bone marrow chromosome aberration test of GHRP 2 in Swiss albino mice indicated that the peptide is non-genotoxic and non-mutagenic up to 2000 mg/kg/day when administered for two consecutive days. Thus, animal studies of general, cytotoxicity and genotoxicity were all negative indicating that GHRP-2 has a high margin a safety. Brief summaries of these reports are provided below as Appendices I - V. Full reports are on file within the archives of ProSoma LLC, Clearwater, FL.
The amino acid leucine and its metabolite ß-hydroxy ß-methyl butyrate (HMB) are included in the Sarcotropin formula because of their ability to increase muscle protein synthesis. Because evidence of toxicity has not been linked to either molecule, there are no bans or restrictions on their in food supplements. While leucine is known to be safe and effective, extremely high doses, well beyond those used in the Sarcotropin formula may cause symptoms of reduced blood sugar as a result of its ability to stimulate insulin secretion (8). Nine studies designed to evaluate HMB safety in which humans found it to be safe for use as an ergogenic (performance enhancing) aid that benefits cardiovascular/general health and improves perception of well being (9). The studies were from 3 to 8 wk in duration, included both males and females, young and old, exercising or non-exercising. Organ and tissue function was assessed by blood chemistry and hematology; subtle effects on emotional perception were measured with an emotional profile test and tolerance of HMB was assessed with a battery of 32 health-related questions. HMB did not adversely affect any surrogate marker of tissue health and function. Furthermore, The effects of HMB on surrogate markers of cardiovascular health suggests that it may decrease the risk of heart attack and stroke.
Four studies that specifically addressed glutamine safety all concluded that glutamine is safe in adults and in preterm infants when taken in doses appropriate for use in food supplements (10).
Mucuna pruriens has demonstrated little toxicity; however, when administered at very high doses to experimental animals it may cause birth defects. Thus, the product should not be used during pregnancy (11).
As one of the safest substances known to man, vitamin D toxicity is very rare. In fact, people are at far greater risk of vitamin D deficiency than they are of vitamin D toxicity (12). However, as with all substances that are capable of affecting bodily functions, extremely high doses can cause toxicity. For example, hypervitaminosis D is a condition that occurs after taking very high doses of vitamin D that causes abnormally high levels of calcium in the blood (13). However, such toxicity occurs at doses more than ten times higher than that found in the Sarcotropin formula.
Toxicities of the inactive ingredients (preservative and water) used in the Sarcotropin formulation have been previously reported and are generally regarded as safe for use at the dosages employed.
Finally, clinical studies were recently completed on the Sarcotropin formulation. The results showed the product effective in its claims and safe for human consumption. Thus, Sarcotropin has been shown to lack side effects and that if any adverse events are reported, they should be of no greater severity or frequency than those occurring spontaneously by chance.
1. Xu Z et al., Ghrelin prevents doxorubicin-induced cardiotoxicity through TNF- alpha/NF-kappaB pathways and mitochondrial protective mechanisms. Toxicology 247:133-138, 2008.
2. Jahromi MG et al. Protective effect of ghrelin on acetaminophen-induced liver injury. Peptides 31:2114-2117, 2010.
3. Slomiany BL, Slomiany A. Ghrelin protection against cytotoxic effect of ethanol. Int J Biomed Sci 6(1):37-44, 2010.
4. Takashi A et al. Pharmacokinetics, safety and endocrine and appetite effects of ghrelin administration in young healthy subjects. Euro J Endocrinol 150(4):447- 455, 2004
5. Strasser F et al. Safety, tolerability and pharmacokinetics of intravenous ghrelin for cancer-related anorexia/cachexia: a randomized, placebo-controlled, double- blind, double-crossover study. Brit J Cancer 98:300-308, 2008.
6. Mericq V et al. Effects of eight months treatment with graded doses of a growth hormone (GH)-relkeasing peptide in GH-deficient children. J Clin Endocrinol Metab 83:2355-2360, 1998.
7. Ghrelin receptor agonist: Executive Summary; RaQualia Pharma Inc. http://www.raqualia.com/portfolio/ghrelin.html
8. Koopman R, Verdijk L, Manders RJ, Gijsen AP, Gorselink M, Pijpers E, Wagenmakers AJ, vanLoon LJ. Co-ingestion of protein and leucine stimulates muscle protein synthesis rates to the same extent in young and elderly lean men. Am J Clin Nutr. 2006; 84:623–632. [PubMed: 16960178]
9. Nissen S, Sharp RL, Panton L, Vukovich M, Trappe S, Fuller JC Jr. b-Hydroxy-b- Methylbutyrate (HMB) Supplementation in Humans Is Safe and May Decrease Cardiovascular Risk Factors . J. Nutr. 130: 1937–1945, 2000.
10.Garlick PJ. Assessment of the Safety of Glutamine and Other Amino Acids. J. Nutr, 131 (9): 2556S-2561S, 2001
11.Rainforest Tropical Plant Database for Velvet Bean (Mucuna pruriens), http://rainforest-database.com/plants/velvetbean.htm
12.Vitamin D Council: http://www.vitamindcouncil.org/about-vitamin-d/what-is- vitamin-d/vitamin-d-toxicity/
13. PubMed Health. A.D.A.M. Medical Encyclopedia, 2011 http://www.ncbi.nlm.nih.gov/pubmedhealth/PMH0002561/
APPENDIX I: Acute Oral Toxicity Study (LD50) of GHRP 2 on Wistar Rats
Based on the observations obtained from this study the GHRP 2 is found to be safe up to 2000 mg/kg when administered as two divided dose within one hour interval to Wistar rats hence the formulation is declared to be in GHS-5/unclassified category.
APPENDIX II: 90 Day Oral Toxicity Study of GHRP 2 on Wistar Rats
In conclusion, when the Wistar rats were exposed to repeated oral administration of GHRP 2 for 90 days, there was no adverse effect observed on any parameters studied such as clinical signs, mortality, body weight, feed consumption, haematology, biochemistry, urine analysis, organ weight, gross pathology and histopathology. Therefore it is concluded that oral administration of GHRP 2 up to 50 mg/kg is found to be well tolerated and under the conditions of this study, the No Observed Adverse Effect Level (NOAEL) for the test article is found to be 50 mg/kg.
APPENDIX III: Mutagenicity study of GHRP 2 by bacterial reverse mutation assay
The values of vehicle control in all the tester strains were within acceptable limit. The positive control revealed a clear increase in number of revertants both in the presence (30% v/v S9 mix) and in the absence of metabolic activation with respective known mutagens when compared with the vehicle control. This demonstrated the efficiency of the test system and suitability of the procedures employed in the study. From the data of the study, it is concluded that GHRP 2 up to dose of 2 mg/mL both in the presence (30% v/v S9 mix) and in the absence of metabolic activation, is non mutagenic to all strains of Salmonella typhimurium viz. TA98, TA100, TA1535 and TA1537 and E.coli strain wp2 [pKM101] and wp2 uvrA when tested under specified condition.
APPENDIX IV: Micronucleus Test of GHRP 2 in Swiss albino mice
The results of mammalian bone marrow micronucleus study of GHRP 2 in Swiss albino mice indicated that the GHRP 2 has not revealed any evidence of causing chromosome damage in this in vivo test and hence it is concluded that GHRP 2 is non-genotoxic or non mutagenic up to 2000 mg/kg/day when administered for two consecutive days.
APPENDIX V: Mammalian Bone Marrow Chromosome Aberration Test of GHRP 2 in Swiss albino mice
The results of mammalian bone marrow chromosome aberration test of GHRP 2 in Swiss albino mice indicated that the GHRP 2 has not revealed any evidence of causing chromosome aberration in this in vivo test and hence it is concluded that GHRP 2 is non-genotoxic or non-mutagenic up to 2000 mg/kg/day when administered for two consecutive days.
APPENDIX VI: A Phase III, Randomized, Open Label, Placebo-Controlled Clinical Study To Evaluate The Efficacy And Safety of Sarcotropin® For The Treatment of Age Related Changes in Form, Function and Quality of Life In Healthy Subjects
Total of 94 male and female healthy subjects were enrolled in the study, out of that 53 subjects were male (56.38 %) and 41 subjects were female (43.62 %). Of these, 75 subjects have completed study successfully. Out of 75 subjects, 50 subjects (M - 29 and F - 21) from treatment groups who received Sarcotropin® where as 25 subjects (M - 13 and F - 12) from placebo groups who received placebo (Sarcotropin® without GHRP). 19 subjects were discontinued from the study due to their personal reason and/or adverse events from the study. The mean age of all subjects received Sarcotropin® treatment was found to be in a range of 40 to 68 years with a mean of 48.89 ± 0.992 years, mean height of subjects was 163.56 ± 1.25 cms and mean weight at 71.67 ± 2.07 kg. The mean age of all subjects received Placebo treatment was found to be in a range of 40 to 62 years with a mean of 49.50 ± 1.24 years, mean height of subjects was 159.57 ± 4.02 cms and mean weight at 72.60 ± 3.86 kg.
Safety: A total of twenty episodes of adverse events were reported out of 94 enrolled subjects. All reported events were resolved. Out of these, five subjects were discontinued from the study. No clinical significant changes were observed in hematology parameters except total platelet count which was higher in the treatment group. However, the values were within the normal range and thus, the changes are considered clinically irrelevant. There were no clinically significant changes observed in biochemical parameters in the treatment group as compared to placebo group.