Single Dose LD50 Safety Test

Title:
Acute Oral Toxicity Study of GHRP-2 in Wistar Rats

Sponsor:
ProSoma, LLC
34911 US Hwy 19N
Suite 525 E
Palm Harbor, FL 34684

Test facility:
Pre-Clinical Department
Contract Research Organization,
Cadila Pharmaceuticals Limited
1389, Trasad Road,
Dholka - 387 810 Dist: Ahmedabad,
Gujarat, India

Study Date: August 19, 2011

Test guidelines:
OECD Guideline for Testing of Chemicals (No. 420, Section 4: Health Effects) on conduct of "Acute Oral Toxicity - Fixed Dose Procedure" (Adopted: 17th December 2001) and in accordance with the mutually agreed study plan.

Good laboratory practice: The study was conducted following the principles of Good Laboratory Practice (GLP) as set forth in the OECD Principles of Good Laboratory Practice, OECD, 1998.

Objective:
The objective of this study was to assess the toxicity of high dose GHRP 2 oral administration to female rats. Acute toxicity studies in animals are usually necessary for any pharmaceutical intended for human use. The information obtained from these studies is useful in choosing doses for repeat-dose studies, providing preliminary identification of target organs of toxicity. Acute toxicity studies may also provide information relevant to acute overdosing in humans. Results of this study also enable ranking of the test material according to Globally Harmonized System (GHS), currently in use.

Materials and methods:
Twenty female rats of 10 weeks age were selected after veterinary examination for the study. Four animals for the sighting study were acclimatized for 5 days, whereas 16 animals intended for the main study were acclimatized for 8 days before initiation of treatment.

Sighting study: The test article (GHRP 2) was administered at a dose of 300 mg/kg to 1 animal, which was observed for one day thereafter. This animal did not reveal any clinical signs during the 24 hour observation period. Hence a second animal was dosed at 2000 mg/kg and observed for one day. This animal did not reveal any clinical signs during entire observation period of 14 days.

Main study: Since 2000 mg/kg did not reveal any toxicity in the sighting study, the remaining animals were administered the same dose and observed for 14 days. After completion of the observation period all animals were euthanised with carbon dioxide gas and then were the subject to necropsy.

Clinical findings:
All animals were observed at ½, 1, 2, 4, 6 and 24 hours after dosing and thereafter once daily once for 14 days to determine if the test materials was associated with any clinical signs of toxicity. The observations included changes in skin and fur, eyes and mucous membranes and also respiratory, circulatory, autonomic and central nervous systems, and somatomotor activity and behavior patterns, tremors, convulsions, salivation, diarrhea, lethargy, sleep and coma. No abnormal clinical signs were observed throughout observation period in both sighting and main test.

Mortality:
There was no deaths observed in any of the animals used for the sighting and main study.

Body weight:
The weight of each animal was recorded on day 0, before dosing (day 1) and at weekly intervals throughout the period of study. No abnormal changes in body weight gain of animals were observed.

Necropsy findings:
No gross pathological changes were observed in any of the animals used for the sighting or main studies.

Conclusion:
Based on the observations obtained from this study, 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. Based upon an average weight of 300 grams per rat, the dose of GHRP-2 administered was 600 mg/rat. In contrast, the human dose of GHRP-2 contained in Sarcotropin is 4 mg/person. Assuming an average body weight of 60 kg (132 lbs), the dose to humans is 0.06 mg/kg or 1/10,000 of that administered to the test animals. As a result of these findings, the peptide fits into the GHS-5/unclassified category (1).

A full copy of this Acute Toxicity Study is on file in the archives of ProSoma, LLC, Palm Harbor, Florida, USA

Literature Cited
1. US Department of Labor, OSHA, A Guide to The Globally Harmonized System of Classification and Labelling of Chemicals (GHS). http://www.osha.gov/dsg/hazcom/ghs.html#3.2