Safety StudyAn initial safety study was conducted on MEP enrolling 60 females ages 53-80 years who had been amenorrheic for at least 3 years with Rao-Goldman wrinkle classification 3+, Glogau aging classification II-IV, and Fitzpatrick skin type I-IV. Subjects were randomized in a 4:1:1 ratio of MEP plus vehicle to vehicle alone to control moisturizer. The study products were applied twice daily to the entire face for 12 weeks. Blood samples were drawn at baseline and week 12 to obtain plasma for the detection of MEP and the inactive metabolite of MEP (MicroConstants, Inc, San Diego, CA).No adverse events or safety issues arose during the conduct of the study. The investigator and subject assessed excellent tolerability for MEP plus vehicle. Subjects receiving active topical MEP rapidly converted the soft estrogen to an inactive serum metabolite alkyl carboxylate E161,0,8 insuring the safety of the novel ingredient.
METHODS
The subsequent efficacy study enrolled 80 female subjects age 53-80 years who had been amenorrheic for 3-10 years with Rao-Goldman wrinkle classification 3+, Glogau aging classification II-IV, and Fitzpatrick skin type I-IV who completed informed consent (Concordia IRB, Cedar Knolls, NJ). Subjects did not use HRT currently or in the past. No oral retinoids or topical products containing a retinoid, retinol, or other vitamin A derivative were allowed. Subjects also did not use systemic corticosteroids or any topical facial medications. Facial dermabrasion, microdermabrasion, laser treatments, chemical peels, botulinum toxin or filler/biostimulatory facial injections were not allowed during the study. Topical sunless tanning products, facial waxing, depilatories, alpha-hydroxy acids, salicylic acid, or vitamin C containing products were forbidden.The investigator and the subjects evaluated overall global facial aging, in addition to a variety of visual signs of facial agingincluding skin thickness, fine lines, wrinkles, dryness, telangiectasias, laxity, atrophy, dullness, and erythema. The investigator queried the subjects and the subjects also evaluated study product tolerability based on burning and itching. All evaluations were conducted on a 5-point scale: 0=none, 1=minimal, 2=mild, 3=moderate, 4=severe. These study activities occurred at baseline, week 6, week 10, and week 14.Successfully enrolled subjects were randomized in a 3:1 ratio of MEP plus vehicle to vehicle alone. The vehicle was developed to possess minimal moisturizing qualities to allow a clear independent assessment of MEP skin activity. The constituents of the vehicle were water, glycerin, benzyl alcohol, magnesium aluminum silicate, saccharide isomerate, xanthan gum, phenoxyethanol, and ethylhexylglycerin. This formulation represents thickened water with no occlusive agents to retard transepidermal water loss. Any water attracted by the humectant glycerin would evaporate to the environment. Subjects were instructed to apply the randomized study product to the entire face, avoiding the eyes and mouth, twice daily for 14 weeks. Compliance was determined based on diary entries and product weights.Nine subjects participated in a biopsy sub-study to evaluate the histologic effects of the MEP active on epidermal thickness (H&E, Cockerell Dermatopathology, Dallas, TX), collagen formation (H&E, Cockerell Dermatopathology, Dallas, TX) and estrogen receptor positivity (estrogen receptor stain, Southwestern University, Dallas, TX). A 3 mm biopsy specimen was obtained from the abdomen at baseline utilizing 2% epinephrine plus lidocaine and suture closure. These subjects were instructed to apply the product to the face and abdomen for the entire study duration followed by a repeat of the abdominal biopsy procedure at week 14.Statistical AnalysisThe randomized active and control groups were balanced to insure no statistically significant differences in skin aging or demographics were present at baseline. A Mann Whitney two-tailed t test was used to analyze the nonparametric ordinal investigator and subject efficacy and tolerability data. Comparisons were made to baseline longitudinally for intragroup evaluations utilizing a two-tailed Wilcoxon ranked sign test to determine percent improvement in evaluated facial characteristics. Intergroup analysis between the active and vehicle was calculated as difference from baseline utilizing a two-tailed Mann Whitney. Significance was defined as P less than or equal to 0.05.
RESULTS
Seventy-nine of eighty females successfully completed the study, with one subject who missed the last visit at week 14 for personal reasons unrelated to the study product. No statistically significant differences in tolerability measurements were noted between the MEP and vehicle arms after twice daily use to the entire face and upper lip for 14 weeks.At week 10, there was statistically significant blinded investigator-rated improvement in skin atrophy (P=0.028) and dullness(P=0.001) for the MEP group over the vehicle group as compared to baseline. The investigator noted MEP induced improvement from baseline at week 14 to include dryness (P less than 0.001) and laxity (P=0.001), in addition to atrophy (P=0.003) and dullness(P less than 0.001), as compared to vehicle. The subject data was evaluated by comparing the raw ordinal scores between the MEP and vehicle only group. A statistically significant decrease in skin atrophy (P=0.047) was noted at week 10 with continuing improvement in lines (P=0.014) and atrophy (P=0.020) at week 14 for the MEP over vehicle. These are visual improvementsthat might be expected after applying a topical soft estrogen
