Micronutrient Supplementation for Women With PCO-syndrome - Influence of Nutrition and Physiology on the Development of PCOS-typical Parameters
Phase: NA
Conditions: Polycystic Ovary Syndrome, Infertility, Female, Micronutrient Deficiency
Sponsor: Medical University of Vienna

Ott Studienprotokoll – PROfertil female PCO 5.0 2017-11-16 
 
 
Study protocol 
 
Project title: Micronutrient Supplementation for Women with PCO-syndrome - 
Influence of Nutrition and Physiology on the Development of PCOS-typical 
Parameters   
 
Participants 
 
Investigator:  
Assoc.Prof. Priv.Doz. Dr. Johannes Ott  
 
Participating doctors 
Assoc.Prof. Priv.Doz. Dr. Johannes Ott  
 
Participating clinics 
Medical University of Vienna 
Department of Obstetrics and Gynecology 
Clinical Division of Gynecologic Endocrinology and  
Reproductive Medicine 
Währinger Gütel 18-20 
A-1090 Vienna 
Tel:  + 43 - 1 - 40400 – 28160 
Fax:  + 43 - 1 - 40400 – 28170 
 
 
 

Ott Studienprotokoll – PROfertil female PCO 5.0 2017-11-16 
 
 
Introduction and scientific background 
In the Western world, malnutrition and subfertility have become increasingly common problems 
affecting both men and women. A recent study showed that conforming to nutritional 
recommendations significantly increased the probability of a clinical pregnancy following IVF/ICSI 
(1). They included fruit, vegetables, certain types of meat, fish, whole-grain products and specific 
fats (according to the Netherlands Nutrition Centre Foundation). Another study specifically 
analysed preconceptional consumption of unsaturated fatty acids by women before IVF/ICSI. It 
discovered a positive effect on the embryo’s morphology (2). 
 
The effect of micronutrients on female fertility is currently under investigation. A 2012 review on 
the subject included 13 studies with more than 90,000 female participants and provided no 
generalised recommendations, but allowed the authors to deduce a certain positive effect of 
supplementation with certain micronutrients on female fertility (3). 
 
This current study will test the effect of a currently available micronutrient supplementation in 
women suffering from polycystic ovary (PCO)-syndrome and sterility. The product (PROfertil ® 
female, Lenus Pharma, Seeböckgasse 59, 1160 Vienna, Austria) is based on several studies that 
showed (i) an increased need for these micronutrients of women with a desire to have a child or 
currently pregnant and/or (ii) a positive effect on female fertility. They are collected in the attached 
PROfertil ® female study report (see attachment “PROfertil Studienreport final”. These documents 
summarise the evidence for the contents of the product in detail. PROfertil ® female is a 
micronutrient supplement containing selenium, vitamin E, catechin, glycyrrhizin, co-enzyme Q10, 
folic acid and omega-3 fatty acids. 
 
This study seems of impact from a scientific perspective. Clinical experience with PROfertil ® 
female suggests that patients suffering from PCO-syndrome experience an increase of anti-
müllerian hormone and androgen levels. The reductive effect of typical PCO-seroparameters can be 
explained pathophysiologically: A chronic low-level pro-inflammatory state – both locally in the 
ovary and systemically – appears to be among the central pathophysiological correlates of PCO-
syndrome (4). PROfertil ® female contains a mix of several antioxidative agents. A favourable 
effect on PCO syndrome is expected. Metformin is an anti-inflammatory substance commonly used 
to treat PCO-syndrome. The anti-inflammatory effect is seen as positive (5). This explains why 
women suffering from PCO-syndrome without insulin glucose balance problems react positively to 
treatment with Metformin (6). 
 
 

Ott Studienprotokoll – PROfertil female PCO 5.0 2017-11-16 
 
 
Study aims 
The primary aim of this study is to analyse the effects of three months of treatment with PROfertil 
® female compared to a supplementation with 400μg of folic acid for women suffering from PCO-
syndrome, sterility, anovulation. The study will examine the parameters typical for PCO-syndrome 
(anti-Mullerian hormone, total testosterone, androstendione). The secondary goal is to document 
the individual course of further fertility treatment. 
 
The products being administered to the patients for four weeks are:  
- 
PROfertil ® female (Lenus Pharma GesmbH, Seeböckgasse 59, 1160 Vienna; treatment 
group) – one soft capsule and one pill per day. 
- 
200µg folic acid (Folsäure Kapseln 400µg ®, OTC Produktion und Forschung GmbH, 
Fischergasse 17, 5020 Salzburg; control group) – two capsules per day. 
- 
 
The study will be performed in a double-blinded manner. The medications are unlabeled for both 
groups and will be dispensed in unlabeled blisteres. However, patients could search for the actual 
look of the PROfertil ® female soft capsules and the pills (for example on the internet). Since they 
differ from the folic acid capsules, patients could identify the control medications. The study team 
is aware of the fact that this kind of blinding is not according to standards and could introduce 
some kind of bias. 
 
PROfertil ® female composition overview 
 
Amount per day 
Contents 
New composition 
Soft capsule 
Omega-3 fatty acids 
500 mg 
Pill 
Folic acid  
800 µg 
Selenium 
70 µg 
Vitamin E 
30 mg 
Green-tea extract catechin  
4 mg 
Glycyrrhizin from licorice extract 
12 mg 
Coenzyme Q10 
30 mg 
 
Study hypothesis:  
First hypothesis: A three-month treatment with PROfertil ® female causes a drop of anti-müllerian 
hormone, total testosterone and androstenedione levels. This does not happen if treated with 
400μg folic acid. 
 
Outcome parameters: 
Primary outcome parameters: Primary outcome parameters: anti-müllerian hormone (AMH), total 
testosterone and androstenedione levels 
 

Ott Studienprotokoll – PROfertil female PCO 5.0 2017-11-16 
 
 
Study design:  
Monocentral, prospective randomized double-blinded trial 
 
Participants:  
The participants will be chosen or excluded using the following criteria:  
 
Inclusion criteria:  
1) The patient has been diagnosed with PCO-syndrome using revised 2004 Rotterdam-criteria (7). 
2) The patient is sterile, defined as being unable to become pregnant within a year despite 
unprotected sexual intercourse.  
3) The patient suffers either from oligomenorrhoea (defined as an interval of ≥60 days between 
the last three menstruations) or complete amenorrhoea for at least 90 days.  
4) The patient has given her written informed consent after detailed information on the study by 
medical professionals at the Department of Obstetrics and Gynecology of the Medical University of 
Vienna.  
5) Both partners are at least 19 years old and are younger than 35. The age limit was chosen to 
take physiologically reduced fertility beyond this age (8). This measure allowed the women to avoid 
a 100-day delay of fertily treatments due to this study.  
 
Exclusion criteria:  
1) No informed consent.  
2) At least one partner is younger than 19 years or older than 35 years.  
3) The patient has been subject to one of the following PCO-syndrome-related treatments within 
three months before inclusion: metformine, combined oral contraceptives, cortisol therapy, inositol, 
ovarian drilling, any kind of ovarian stimulation, in-vitro fertilisation. Promoting menstruation using 
gestagen products is acceptable.  
 
The inclusion and exclusion criteria are checked at the start of the study and must be fulfilled at 
that point. All inclusion criteria have to be fulfilled to participate. A single exclusion criterion is 
sufficient to prevent participation.  
 
Recruitment:  
The participants are recruited by medical professionals at the Department of Obstetrics and 
Gynecology of the Medical University of Vienna using the above mentioned criteria during routine 
examinations before any treatment for PCO syndrome and/or sterility is initiated. Potential 
participants are informed about the procedure, clinical relevance and possible additional effort 
caused by study participation. The patients willing to participate then have to sign the written 
informed consent. 
 
Sample size calculation:  
Assuming a reduction of the mean AMH levels by 2 ng/ml at a standard deviation of 3 ng/ml and 
an alpha value of 0.05 and a power of 0.90, the paired t-test requires 26 patients per group. Due 

Ott Studienprotokoll – PROfertil female PCO 5.0 2017-11-16 
 
 
to the fact that the participants wish to conceive a child and have been diagnosed with sterility, a 
low drop-out rate can be expected. There is no reliable data on this or similar treatments that allow 
prediction of the chance to conceive. Due to the favourable benefit-to-risk profile, a 15% drop-out 
rate is assumed. This corresponds to 4 patients per group. Thus, the final sample size is calculated 
as 30 patients per group, which leads to a total study population of 60. 
 
Statistical analysis:  
The bio-statistician will randomise the data using nQuery advisorTM Version 7.0. The information 
will then be packaged in an envelope by a person independent of the participating medical 
professionals and other scientific personnel. This will only be opened after the inclusion of the 
patient. Categorical variables will be presented as absolute numbers and percentages, numerical 
variables as median and interquartile range. 
 
The following statistical analyses are planned: The result parameters and the patient characteristics 
of the two groups will be compared using the Welch test (for numerical variables) and the Chi-
square-test or Fisher’s exact test (for categorical variables). A p-value of <0.05 is considered to be 
statistically significant. The analysis will be performed as an intention-to-treat analysis. The 
statistical analyses will be performed using SPSS 24.0 for Windows (SPSS Inc., 1989-2017). 
 
Overview on parameters to be analysed:  
Parameter 
Unit 
Type 
Planned statistical test 
Age 
Years 
Numerical 
Welch test 
Body mass index 
Kg/m2 
Numerical 
Welch test 
Sterility: Secondary 
(versus primary) 
n (%) 
Categorical 
Chi-square/Fisher’s exact test 
Duration of infertility 
Years 
Numerical 
Welch test 
Pre-treatment FSH 
mU/mL 
Numerical 
Welch test 
Pre-treatment LH 
mU/mL 
Numerical 
Welch test 
Pre-treatment AMH 
ng/mL 
Numerical 
Welch test 
Pre-treatment 
testosterone 
ng/mL 
Numerical 
Welch test 
Pre-treatment 
androstenedione 
ng/mL 
Numerical 
Welch test 
Post-treatment AMH 
ng/mL 
Numerical 
Welch test 
Post-treatment 
testosterone 
ng/mL 
Numerical 
Welch test 
Post-treatment 
androstenedione 
ng/mL 
Numerical 
Welch test 
Abbreviations used: n (%) = number (frequency) 
 
 
 

Ott Studienprotokoll – PROfertil female PCO 5.0 2017-11-16 
 
 
Further details on study design:  
After recruiting the participants at the outpatient clinics for Gynecologic Endocrinoloy and Infertility 
Treatment, they are randomly assigned to one of the groups. Participants will receive either 2 
unlabelled soft capsules containing 200μg folic acid each or 1 unlabelled soft capsule containing 
omega-3 fatty acids and 1 tablet containing folic acid, selenium, vitamin E, catechines, glyzyrrhicin, 
and co-enzyme Q10 (see Table above for details on doses). 
The planned examination of AMH, total testosterone and androstenedione levels will take place 
after a minimum of 90 days (and a maximum of 100 days) after beginning supplementation with 
PROfertil ® female or folic acid. The maximum duration of participation is thus 100 days. After 
taking the micronutrient supplementation (PROfertil ® female or folic acid) beyond the planned 
minimum treatment duration, an individualised fertility treatment can be performed. 
There are two study-specific consultations: After including the patient (consultation 1: 
randomisation, distribution of drugs and start of study) and after 90 to 100 days (consultation 2). 
During consultation 2, unused study blisters are collected and patients are questioned on 
peculiarities and possible side effects of the micronutrient supplementation used. 
It is expected that 30 minutes for consultation 1 and 15 minutes for consultation 2 will have to be 
spent by the patient. The consultations will be held during regular outpatient treatments and will be 
part of the planned routine examinations. They include discussions of examination results and 
recommendations for further treatment. 
 
Study duration:  
A total duration of about 12 months will not be exceeded.  
 
Benefits-to-risk assessment and preventive measures:  
Risks or complications due to the use of PROfertil ® female and additional venepuncture are not 
expected. A recent study using the same product showed no substance-related side effects 
(number of the ethics committee of the Medical University of: 1659/2013; accepted on 2013-09-
24) (9). A direct benefit for the treatment group is possible. A positive effect on PCO syndrome can 
be assumed, which might result in a higher chance of pregnancy. The substance administered to 
the control group is state of the art for female fertility treatment. 
• 
Fertility treatment can be postponed by a maximum of 100 days by this study. This 
information is communicated to the patients during informed consent. Should there be any 
unexpected complications or should the patient want to stop her participation for personal 
reasons, she will be excluded and will no longer be subject to further examinations.  
• 
It is possible for the patients to benefit financially from this study since Lenus Pharma 
GesmbH provides them with the study medication (PROfertil ® female and Folsäure 
Kapseln 400µg). In addition, the company will provide the assigned medication free of 
charge for further 6 months after the end of the study, should they wish to continue 
treatment for PCO-syndrome and fertility.  
 
Study-specific parameters:  
None. 

Ott Studienprotokoll – PROfertil female PCO 5.0 2017-11-16 
 
 
 
Parameters obtained during routine clinical examinations:  
All parameters on outcome (anti-müllerian hormone, total testosterone and androstendion levels) 
will be obtained during routine clinical examinations.  
Several additional parameters will also be collected and included in the database. They do not 
require additional effort on behalf of the patients. They include: general patient information (age, 
duration of infertility, height and body weight, additional medication); information pertaining to 
PCO-syndrome (menstruation history, Ferriman-Gallwey-score on hirsutism (if applicable), acne, 
scalp-hair effluvium); hormone parameters on days 2 to 5 of the menstrual cycle (androstendion, 
thyroid-stimulating hormone, luteal hormone, follicle-stimulating hormone, sex hormone binding 
globulin); as well as an oral glucose tolerance test, which is routinely performed by general 
practicioners.  
In addition, the individual development of the fertililty treatment will be documented for a 
maximum of 6 months following active study participation.  
 
 

Ott Studienprotokoll – PROfertil female PCO 5.0 2017-11-16 
 
 
Approval of PROfertil ® female: 
PROfertil ® female is approved in Austria by the Austrian Ministry of Health. A copy of the 
corresponding Free Sales Certificates can be found in the attachment.  
 
Approval of 400µg Folsäure 400: 
Folsäure Kapseln 200µg (OTC Produktion und Forschung GmbH ®) is a nutritional supplement. 
Contrary to drugs, these products do not require approval, registration or declaration in Austria.  
 
Study financing: 
Study medication and ongoing medication were provided by Lenus Pharma GesMbH (Seeböckgasse 
59, 1160 Vienna, Austria). Lenus Pharma GesMbH supports clinical trial expenditures with a total 
sum of EUR 9,000.00 for 60 participants. Lenus Pharma GesMbH is not a direct sponsor of the 
study. The data and usage rights remain with the Medical University of Vienna.  
 
Data access agreement: 
Assoc.Prof. Priv.Doz. Dr. Johannes Ott can access all data and statistics at any time.  

Ott Studienprotokoll – PROfertil female PCO 5.0 2017-11-16 
 
 
Literature: 
1. Twigt JM, Bolhuis ME, Steegers EA, Hammiche F, van Inzen WG, Laven JS, Steegers-Theunissen 
RP. The preconception diet is associated with the chance of ongoing pregnancy in women 
undergoing IVF/ICSI treatment. Hum Reprod. 2012;27:2526-31. 
2. Hammiche F, Vujkovic M, Wijburg W, de Vries JH, Macklon NS, Laven JS, Steegers-Theunissen 
RP. Increased preconception omega-3 polyunsaturated fatty acid intake improves embryo 
morphology. Fertil Steril. 2011;95:1820-3.  
3. Grajecki D, Zyriax BC, Buhling KJ. The effect of micronutrient supplements on female fertility: a 
systematic review. Arch Gynecol Obstet. 2012;285:1463-71. 
4. Spritzer PM, Lecke SB, Satler F, Morsch DM. Adipose tissue dysfunction, adipokines, and low-
grade chronic inflammation in polycystic ovary syndrome. Reproduction. 2015;149:R219-27. 
5. Sirmans SM, Weidman-Evans E, Everton V, Thompson D. Polycystic ovary syndrome and chronic 
inflammation: pharmacotherapeutic implications. Ann Pharmacother. 2012;46:403-18. 
6. Onalan G, Goktolga U, Ceyhan T, Bagis T, Onalan R, Pabuçcu R. Predictive value of glucose-
insulin ratio in PCOS and profile of women who will benefit from metformin therapy: obese, lean, 
hyper or normoinsulinemic? Eur J Obstet Gynecol Reprod Biol. 2005;123:204-11. 
7. Thessaloniki ESHRE/ASRM-Sponsored PCOS Consensus Workshop Group. Consensus on 
infertility treatment related to polycystic ovary syndrome. Fertility and Sterility 2008;89:505-22. 
8. See also: American Society for Reproductive Medicine. Age and Fertility – A Guide for Patients. 
Verfügbar 
online 
unter: 
https://www.asrm.org/uploadedFiles/ASRM_Content/Resources/Patient_Resources/Fact_Sheets_an
d_Info_Booklets/agefertility.pdf 
9. Nouri K, Walch K, Weghofer A, Imhof M, Egarter C, Ott J. The Impact of a Standardized Oral 
Multinutrient Supplementation on Embryo Quality in in vitro Fertilization/Intracytoplasmic Sperm 
Injection: A Prospective Randomized Trial. Gynecol Obstet Invest. 2017;82:8-14.