AEOL 10150 Development

Phase I Single Dose Study - COMPLETED:

Data presented here are unaudited and therefore subject to change upon complete review. If changes need to be made they will be and a similar notice will be posted.

AEOL 10150 Study 101 – A Double-Blind, Randomized, Placebo-Controlled, Phase 1, Escalating Single Dose Study to Evaluate the Safety, Tolerability and Pharmacokinetics of AEOL 10150 Administered by Subcutaneous Injection to Patients with Amyotrophic Lateral Sclerosis.

Initial Human Dose

The initial dose selected for the first study of AEOL 10150 in humans was 3 mg.

An initial human dose of 3 mg (0.05 mg/kg in a 60 kg subject) is expected to be well tolerated because it is much smaller, on a mg/kg basis, than the doses that were free of adverse effects in three species of animals. The lowest no observed adverse effect level (NOAEL) in the rat after subcutaneous administrations of AEOL 10150, based on findings in a single dose cardiopulmonary safety pharmacology study, was 5 mg/kg. Adverse effects observed with higher single doses of 12.5 and 25 mg/kg in this same study in the rat were decreased body temperature and dose related decreases in blood pressure, heart rate and respiratory rate. The lowest NOAEL in the mouse, based on findings in the 28-day toxicology study, was 20 mg/kg/day. Finally, the lowest NOAEL in the cynomolgus monkey, based on findings in a single dose cardiopulmonary safety pharmacology study, was 2.7 mg/kg.

In December 2002, the FDA published a Draft Guidance for Industry and Reviewers entitled: “Estimating the Safe Starting Dose in Clinical Trials for Therapeutics in Adult Healthy Volunteers.” This guidance suggests the conversion of the NOAEL of the most sensitive species to a human equivalent dose (HED) by normalizing doses to body surface areas and then the application of an additional safety factor (the default being a 10-fold reduction). Normalizing for body surface area, the HEDs of the lowest NOAELs for each of the rat, cynomolgus monkey and mouse are:

• rat: 0.80 mg/kg (5.0 mg/kg • 0.16)
• monkey: 0.86 mg/kg (2.7 mg/kg • 0.32)
• mouse: 1.6 mg/kg (20 mg/kg • 0.08)

Therefore, the most sensitive species for AEOL 10150 is the rat, for which the HED of the lowest NOAEL is 0.80 mg/kg. Dividing the HED by a safety factor of 16 gives 0.05 mg/kg, which for a 60 kg healthy subject translates to a single dose of 3 mg. Since AEOL 10150 is the first in a new class of compounds and the cardiovascular effects observed in rats and cynomolgus monkeys had a somewhat steep dose-dependency, a safety factor greater than 10 was used to establish the safe starting dose for the first human trial.

In AEOL Study 101, the escalations in dosages (in sequential groups of five subjects each) were from 3 mg to 12 mg to 30 mg to 45 to 60 mg to 75 mg, representing sequential increases in dosage of 300%, 150%, 50%, 33%, and 25%.

The highest planned dosage of 75 mg was established by the formulation of study drug that contains 75 mg/mL of AEOL 10150 and the physical limitation of not wanting to administer more than 1 mL of study drug product by subcutaneous injection. A dose of 75 mg (or 1.25 mg/kg in a subject weighing 60 kg) is 45 to 56% higher than the HED to the lowest NOAELs in rats and cynomolgus monkeys (0.8 m/kg and 0.86 mg/kg, respectively), that were defined by effects on respiratory rate, blood pressure or heart rate, and approaches the HED of 1.6 mg/kg that did demonstrate adverse effects in these species.

The human equivalent to the 2.5 mg/kg once-daily, subcutaneous dose that demonstrated efficacy in the transgenic mouse model of ALS is 12 mg (in a 60 kg individual). Therefore, the six planned dosages of 3 mg, 12 mg, 30 mg, 45mg, 60 mg and 75 mg represent doses that are 0.25, 1, 2.5, 3.5, 5 and 6.25 times the once-daily, subcutaneous dose that demonstrated efficacy in the transgenic mouse model of ALS, respectively.

Summary of Results

In the study, 4-5 patients diagnosed with ALS were utilized in each dosage cohort (3 or 4 receiving AEOL 10150 and 1 receiving placebo). Each dose cohort was evaluated at a separate clinical center. In total, seven separate cohorts were evaluated for the study, and 25 ALS patients received AEOL 10150.

Based upon an analysis of the data, it was concluded that single doses of AEOL 10150 ranging from 3 mg to 75 mg were well tolerated. In addition, no serious adverse clinical events were reported, nor were there any significant laboratory abnormalities.

Based upon extensive cardiovascular monitoring (i.e. frequent electrocardiograms and continuous Holter recordings for up to 48 hours following dosing), there were no compound-related cardiovascular abnormalities.

Following administration of single doses of AEOL 10150 (3, 12, 30, 45, 60 and 75 mg), pharmacokinetic analysis demonstrated plasma area under the curve (AUC) values ranging from 354 ng-hr/mL in the 3 mg group to 12,167 ng-hr/mL in the 75 mg group. Correspondingly, Cmax ranged from 114.8 ng/mL to 1584 ng/mL, and Tmax ranged from 1 to 2 hours in these same groups. The mean half-life of AEOL 10150 ranged from 2.6 (3 mg cohort) to 6.4 hours (75 mg cohort). Linear dose response, and dose proportionality, were documented. A summary of these results is provided below in table form below:

Pharmacokinetic Parameters for AEOL 10150:
Result Summary

The most frequently reported adverse events were injection site reactions, followed by dizziness and headache. Please see the sections below, “Adverse Events” and “Injection Site Reactions” for further information.

Taken together, the pharmacokinetic data indicate that accumulation of AEOL 10150 with multiple dosing is unlikely.

Summary of Plasma Concentration over Time

The graph below summarizes the data developed form the single dose study of AEOL 10150 based upon cohort and size; it is noted that dose proportionality was observed, despite the compromised nature of the patients in the study, as well as the use of a variety of medications (see section below on “Concomitant Medications.”)

Concomitant Medications Taken During Study

Three placebo patients and 20 AEOL 10150 patients took concomitant medications. The dosages and reasons for administration are listed below:

Downlaod Table: Cocontaminant Mediactions Taken During Study

Adverse Events

Among patients who received study medication, adverse events were reported for 92% (24 of 26) of the AEOL10150 patients and 71% (5 of 7) of the placebo patients. Over all of the AEOL10150 groups, events with the greatest incidences were injection site reactions (92%) and headache (81%). Adverse events were primarily mild in severity, with 89% of the individual events being reported as mild (90 of 111 events). No serious adverse events were reported.

All non serious and serious adverse events (SAEs), regardless of suspected causal relationship to study participation, were summarized by treatment arm. Adverse events for the single injection site on each individual patient were considered a single adverse event (Injection site reaction) even if separate entries were made for each description, i.e., erythema, edema, etc. The summary and percentages are listed below:

Downlaod Table: Adverse Event Findings

Injection Site Findings

Injection site findings were not considered adverse events unless they were reported with the AEs but they were summarized by cohort, finding, and time point.

Findings were categorized into:

• Stinging, Burning or Itching
• Discoloration (note color)
• Indurations
• Edema
• Erythema
• Bleeding
• Infection (pus formation)
• Tenderness
• Other (describe)

and were further categorized by severity:

• None (0)
• Slight (1)
• Mild (2)
• Moderate (3)
• Severe (4)

These findings were noted predose, Hour 1, Hour 6, Hour 24, Hour 48, and Day 7. Postdose, there were fifty (50) notable findings above the None (0) level, eighteen (18) of which did not have a severity rating. For further analysis the unrated findings were rated based on the text written with the finding. The findings are summarized below:

Downlaod Table: Injection Site Findings

Phase I Multiple Dose Study -- COMPLETED:

AEOL 10150 Study 102 – A Double-Blind, Randomized, Placebo-Controlled, Phase 1, Escalating Multiple Dose Study to Evaluate the Safety, Tolerability and Pharmacokinetics of AEOL 10150 Administered by Subcutaneous Injection to Patients with Amyotrophic Lateral Sclerosis.

In October 2006, we completed a multi-center, double-blind, randomized, placebo-controlled, Phase Ib clinical trial. This multiple dose study was conducted to evaluate the safety, tolerability and pharmacokinetics of AEOL 10150 administered by subcutaneous injection and infusion pump in patients with ALS. Under the multiple dose protocol, three groups of six ALS patients (four receiving AEOL 10150 and two receiving placebo) were enrolled, based upon patients who meet the El Escorial criteria for Clinically Definite ALS, Clinically Probable ALS, Clinically Probable-Laboratory Supported ALS, or Definite Familial-Laboratory Supported ALS (i.e., Clinically Possible ALS with an identified SOD gene mutation) .

The first two cohorts of the Phase Ib multiple dose study received a fixed daily dose of AEOL 10150 twice a day by subcutaneous injection. In the first cohort, each patient received twice daily subcutaneous injections of 40 mg of AEOL 10150 or placebo, for six consecutive days, followed by a single subcutaneous injection on the seventh day, for a total of 13 injections. In the second cohort, each patient received twice daily subcutaneous injections of 60 mg of AEOL 10150 or placebo, for six consecutive days, followed by a single subcutaneous injection on the seventh day, for a total of 13 injections.

In contrast, the third cohort received a weight adjusted dose (i.e. mg per kilogram of body weight per day) delivered subcutaneously over twenty four hours by continuous infusion pump. In the third cohort, each patient received AEOL 10150 via continuous infusion pump for six and one half consecutive days for a total of 2.0 mg per patient kilogram per day. Each patient in all three cohorts completed the study and follow-up evaluation at 14 days.

The Phase Ib study was conducted at five academic clinical ALS centers. Male and female ALS patients, 18 to 70 years of age, who were ambulatory (with the use of a walker or cane, if needed) and capable of orthostatic blood pressure assessments were enrolled in the study. Clinical signs/symptoms, laboratory values, cardiac assessments, and pharmacokinetics (PK) were performed and currently are being analyzed. The final clinical study report is expected to be released in January 2007.

Based upon a preliminary analysis of the data, it was concluded that multiple doses of AEOL 10150 for a period of six and one half consecutive days in the amount of 40 mg per day, 60 mg per day and 2 mg per kilogram per day were safe and well tolerated. No serious or clinically significant adverse events were reported or observed. The most frequent adverse events related to study compound were injection site observations related to compound delivery. There were no significant laboratory abnormalities. Based upon extensive cardiovascular monitoring (i.e., frequent electrocardiograms and continuous Holter recordings throughout the six and one half days of dosing), there were no compound-related cardiovascular abnormalities.

The pharmacokinetic results of the Phase Ib multiple dose study are being analyzed and modeled. Preliminary results for the first two cohorts are presented below.

Pharmacokinetic Parameters for AEOL 10150 for Phase I Multiple Dose Evaluation (Cohorts 1 and 2).

Pharmacokinetic findings from the Phase Ib study to data are as follows:

• Increases in Cmax and AUC(0-8) appears to correlate with increases in dose, but the correlation is not strong.
• The mean Cmax for the 40 mg cohort was 1,735 ng/mL; and 2,313 ng/mL for the 60 mg cohort.
• There were linear correlations between both Cmax and AUC(0-8) and dose based on body weight.
• The terminal half life (a measurement of the time period for which a compound stays in the body) as determined from Day 7 data was approximately 8 to 9 hours. The shorter (i.e. 4-5 hours) half life determined from Day 1 data is most probably related to distribution of compound.
• Steady-state occurs within three days of multiple dosing. There was no evidence for a third longer half life that would be associated with long term accumulation. Thus, importantly compound accumulation is not expected beyond the third day with multiple dosing.

We are currently working on launching a new clinical trial for AEOL 10150 to further test the safety and efficacy of the drug candidate. The target indication will be in radiation protection with the specific tumor type still being investigated. In addition, we are analyzing the results of the Phase I multiple dose study to determine the appropriate dose level for the proposed study. We expect to file an IND with the FDA during the first half of 2007 and launch this trial in the second half of fiscal year 2007.