BMP INC.: ANIMAL & HUMAN SAFETY OF LYM-X-SORB™ (LXS™)

THIS SUMMARY WAS INCLUDED IN A SBIR PHASE II GRANT APPLICATION ON APRIL 1, 2005

2. LYM-X-SORB Supplement Safety Data 1987 to 2001:

Animal Studies:
A single dose of the LYM-X-SORB™ eutectic was administered at dose volumes of 2.5, 5.0 or 10.0 ml/kg to groups of 10 male rats and a separate group was used as controls (McNeil Pharmaceutical, Spring House, PA 1987). The growth of rats (3, 7 and 14 days) administered LYM-X-SORB™ at 2.5 and 5 ml/kg was comparable to the control group. The largest dosed animals did show reduced growth. The only clinical sign in the 2.5 and 5 ml/kg treated rats was moist rales. Additional mild clinical observations (unkempt appearance in 8 of 10 rats, chromorhinorrhea in 3 of 10, urine stained abdominal hair in 5 of 10, decreased spontaneous motor activity in 5 of 10, ptosis in 5 of 10, and pale extremities in 1 of 10) were noted for the 10 ml/kg dose.

Individual beagle dogs (4 males and 4 females) were administered a single dose of encapsulated LYM X-SORB™ at 1 .0 ml/kg (9-11 kg) and control dogs (2 of both sexes) were dosed with a comparable number of empty hard gelatin capsules (McNeil Pharmaceutical, Spring House, PA, 1987). Recorded observations were made at 3, 7, 10, and 14 days post dosing. No deaths occurred in either group and weights were maintained for all animals. All LYM-X-SORB™ animals were devoid of any clinical signs.

A morphology study of LYM-X-SORB™ in rats was conducted (Upjohn Company, Kalamazoo Ml, 1990). Four groups of fasted rats received 1 ml of (a) saline, (b) aqueous LYM-X-SORB™ (c) aqueous LYM-X SORB™/drug or (d) 0.1M citric acid by injection into the duodenum through a midline incision. In order to observe immediate effects, portions of the jejunum were removed at 5-20 cm from the site of duodenal injection 15 minutes after administration. Intestinal cross sections were processed histologically. No significant differences were found in epithelial cell integrity after administration of LYM-X-SORB™ or LYM-X-SORB™/drug compared to saline; whereas, 0.1 M citric acid resulted in loss of epithelial cells and other abnormal effects.

A study of multiple dosing to pregnant rats and their male offspring was conducted (Guy Lepage, PhD, and Claude C. Roy, MD of Hopital Sainte Justine, Pediatric Research Center, Universite of Montreal, Montreal (Quebec) Canada, 1995-1996). LYM-X-SORB™ (LYM) was fed as the only source of lipids (22% by weight in a balanced diet) to fifteen pregnant rats a week prior to birth, and thereafter during lactation, also to selected male offspring (fifteen) until four months of age. Two control groups of rats were treated similarly except the source of fat was triglyceride (22% having a fatty acid profile similar to LYM-X-SORB™ (LXS) or 5.5% in commercial chow). Weights of each animal, food intake over 3-4 days and any appearances of abnormal behaviors were noted weekly for each animal. LYM provided the caloric demands of pregnancy and lactation without any adverse effect. There were no statistical differences among the growth of the three groups of male rats and no adverse clinical observations were recorded. All animals were healthy, robust and active.

A 13-week oral toxicity study was conducted (ClinTrials BioResearch, Project No. 87680, Senneville (Quebec) Canada, 1997) to determine the potential chronic toxicity of LYM-X-SORB™ in comparison to triglyceride. A diet consisting of 20% fat (w/w) was administered to four groups of rats (N=20/sex/group). The percentage of LYM-X-SORB™ comprising the total fat concentration in the 4 groups was 0, 5, 10 and 20% and the corresponding levels of triglycerides were 20, 15, 10 and 0% respectively. The toxicological evaluations were unremarkable. There were no treatment-related clinical signs, no effects on body weights, and no deaths. Hematology, clinical chemistry and urinalysis were within normal parameters, as were gross and histological pathology and organ weight evaluations.

Human Studies:
In order to evaluate the extent of absorption of 13C-labeled LYM or triglyceride a crossover design was conducted in subjects with CF and healthy volunteers [SBIR Phase I: 1 R43 OK 48208-01, Guy Lepage, PhD, Claude Roy, MD, of Hopital Sainte Justine, Pediatric Research Center, Universite of Montreal, Montreal (Quebec) Canada, and David W. Yesair, BioMolecular Products, Inc. Byfield MA, 1994]. Decreased respiratory excretion rates of 13CO2 without enzyme supplementation, indicate fat malabsorption. In the absence of pancreatic enzyme medication, the subjects with CF poorly absorbed 13C- labeled triglycerides. However, in the 13C-labeled LYM-X-SORB™ subjects with CF and healthy subjects had equivalent absorption, which was similar to 13-labeled triglycerides in healthy subjects. Thus, LYM-X-SORB™ represents a readily absorbable lipid matrix.

In a crossover design study and in the absence of pancreatic enzyme supplements, the acute oral absorption of LYM-X-SORB™ supplemented with added retinyl palmitate, was compared to that of Scandishake® with added retinyl palmitate (Guy Lepage, PhD, Claude Roy, MD, of Hopital Sainte Justine, Pediatric Research Center, Universite of Montreal, Montreal (Quebec) Canada) (J. Pediatr. 2002;141:178-85). Approximately 29 g/m² of body surface area of LYM-X-SORB™ or triglyceride (Scandishake®) and 48,000 I.U./m² of body surface area of retinyl palmitate were orally consumed at time zero. In five subjects with CF and three controls (all fasted overnight), plasma triglyceride peaked at approximately 2 hours (Cmax) and decreased thereafter until 12 h (area under curve, AUC). Both Cmax and AUC for the LYM supplement were statistically greater by 10-fold than the corresponding values following the ingestion of Scandishake®. Similar 10 fold differences were observed for the absorption of retinyl palmitate. Again, LYM was shown to be a readily absorbable lipid matrix and enhances the absorption of fat-soluble retinyl palmitate.

In a one year double blind feeding study (Guy Lepage, PhD, Claude Roy, MD, of Hôpital Sainte Justine, Pediatric Research Center, Université of Montréal, Montréal (Québec) Canada, 1998-2001) (J. Pediatr. 2002;141:178-85), the daily consumption of LYM-X-SORB™ in comparison to triglyceride (24g of lipid per 1.72 m2 of body surface) by subjects with CF (who also consumed, daily pancreatic enzyme supplement) had improved the clinical outcome. The LYM and triglyceride formulations contained at least 50% polyunsaturated fatty acid with a mole ratio of linolenic and a-linolenic fatty acids of about 5:1. LYM-X-SORB™ contained about 20% (w/w) LPC. Per protocol analysis of 48 subjects showed the LYM-X-SORB™ produced better clinical outcomes in comparison to triglyceride supplements as follows:
Increased energy intake (-10%) from diet (P 0.002)
Increased plasma levels of linoleic acid (P <0.001) and a-linolenic acid (P <0.01)
Increased plasma levels of vitamin E (P <0.001) and retinal-binding protein (P = 0.02)
Increased growth in terms of weight (P <0.05) and height (P = 0.03) Z scores
Improved lung function: FEV1 (P 0.02) and PE max (P = 0.02)

Liver and other clinical biochemistry did not change in the subjects. In conclusion, the readily absorbable LYM was an effective and safe medical food.

Risk for Adverse Events:
In the 12-month Montreal study (J. Pediatr. 2002;141:178-85) of 73 subjects on LYM-X-SORB™ or a triglyceride placebo, adverse event data were collected and included in the final report (Protocol #960160) from Ferndale Labs, Inc. No adverse events were directly attributed to the LYM product. The most common adverse event was abdominal pain and was similar in both the LYM-X-SORB™ and triglyceride groups. Other adverse events were rhinitis, diarrhea, eructation, cough, anorexia, nausea, headache, fever and vomiting, and were similar between the two groups. These symptoms are also common in people with CF. Of the reported adverse events, 81% were described as mild intensity, 18% moderate, 1% severe, and < 1% unknown. There were 42 serious adverse events reported by 22 subjects and similar between groups. All were attributed to a cause other than the study supplement, and generally, were related to the underlying CF. There were no deaths.

In summary, the risk of adverse events to our subjects taking LYM for 18 months is minimal.