Improvement of IVF Fertilization Rates, by the Cyclic Tripeptide FEE - Prospective Randomized Study

Name and address of the coordinating investigator : Prof. Jean Philippe Wolf, Histology - Embryology Department , Biology of Reproduction - . Cochin Hospital , 123 Bd Port Royal, 75014 Paris Promotor: AP- HP represented by DRCD - Hôpital Saint- Louis Inclusion Sites : CHU Cochin, Paris

Main objective:

• To improve fertilization rate in IVF by supplementation of culture media with a molecule increasing the sperm fertilizing ability.

Secondary objectives

• Compare the rates of cleavage between the two groups of zygotes
• Compare the percentage of good quality embryos

Number of patients : 160 Duration of the inclusion period : 15 months Duration of participation of the patient: For each couple, the duration of participation will be as many times nine months that pregnancies will be obtained from fresh or frozen embryo transfers.In the absence of pregnancy duration of participation will be only 15 days.

Time monitoring of children : 9 months

Total study duration : The total study duration will be 36 months

Rational :

The cyclic tripeptide FEEc reproduces the binding site of Fertilin Beta. It binds to the alpha 6 beta 1 integrin and increases sperm fertilizing ability, improving its movement's parameters . Added to insemination IVF media, it should increase fertilization rates .

Methodology:

Oocytes in each cohort will be divided into two groups, one of which will be inseminated with the control medium and the other group is inseminated in the presence of peptide. The fertilization rate and embryo quality will be compared between these two groups.

Statistical analysis:

In mice with healthy gamete, the observed fertilization rate increase in the presence of peptide was 100 %. The fertilization rate in human IVF is about 65%, male indications included. There is an improvement of the fertilization index of zona free human oocytes of about 90% . We can reasonably foresee an improvement of 15 % of the fertilization rate in normal IVF.

This is why we believe that the difference between fertilization rates of both treated and control groups must be meaningful for 160 attempts. In this calculation it was assumed that the intra cluster correlation (ie intra cohort oocyte ) resulted in a 'design effect' D equal to 4.

The analysis will include descriptive statistics for each quantitative parameter at each time : mean, standard deviation , minimum, maximum , median and quartiles , number of missing values . Quality parameters will be expressed by their frequency of distribution and associated 95% bilateral confidence intervals.The distributions of the numbers of blastomeres per embryo will be compared between the two groups by exact permutation test (stratified). This method is more suitable than "naive" comparison of these numbers, since it takes into account the possible correlation between results for embryos belonging to the same embryo cohort (Gibbons JD 1985 Non parametric statistical inference - Marcel Dekker New York ). The cellular index, the pregnancy rate, spontaneous abortions, therapeutic abortion, childbirth, will be compared using Student's t test and exact Fisher test. All tests will be bilateral, and look for possible negative effects, set at 5%. Calculations will be performed using SAS statistical software v9.13 (SAS Institute, Cary , NC) under the supervision of Professor Eric Vicaut URC Lariboisière St. Louis.

Primary endpoint : Rate of fertilization Expected Benefits : Improved fertilization rates of at least 15 %