تركيب وتقييم تحاميل مهبلية بروجيستيرون 200 ملغم المستعمل في عقم النساء

Abstract

This work has been carried out to develop pharmaceutical product contains containing progesterone (200 mg) as suppositories, which is used infertility treatment as a part of assist reproductive system for infertility women unable of pregnancy due to a lack of natural progesterone hormone Several formulations were prepared using different suppositories bases, preservatives and humectants. Selection of the best formula relied solely on physicochemical testing of sample exposed to thermal stresses. Stability study was conducted on the product for (6) months at different temperatures ( 2-8 ◦C,25◦C , 30 ◦C ) to determine the expiration date and the best storage conditions. The results revealed a promised suppositories of good homogeneity, smoothness and hardness . The expiry date calculated to be not less than (3) years.

Key words : Progesterone, Suppositories, infertility.

Introduction Normal ovulating women secrete progesterone during the second half of the menstrual cycle by the corpus luteum which forms from the dominant follicle from which the oocyte has been released. Since the corpus luteum dominates this part of the cycle it is known as the luteal phase. Progesterone induces a secretory transformation of the uterine glands, increases vascularity of the endometrial lining, and stabilizes the endometrium in preparation for embryo implantation. Progesterone is also important in interacting with progesterone receptors on gamma/delta T cells leading to the expression of a protein that interferes with natural killer cells especially at the maternal fetal interphase. [1]. Progesterone is a naturally occurring steroid hormone produced by the adrenals, gonads, nervous system, and placenta in pregnancy. It is a derivative of cholesterol. Cholesterol is converted into pregnenolone by enzyme cytochrome P450, and pregnenolone is converted to progesterone by3β-hydroxy-steroid dehydrogenase,isomerase in the smooth endoplasmic reticulum. Progesterone derives its name from ‘progestational steroid hormone’ due to its primary function of preparing and maintaining the uterine bed for conception.Its benefits in decreasing early pregnancy loss via providing luteal support, both in women with threatened abortion and those undergoing artificial reproduction cycles. [2] Progesterone is available in its natural form or synthetic form. Even though both forms are used for similar purposes, they are not chemically identical and their effect on target organs may not be similar. [2 ,3] Treatment with compounded vaginal suppositories has been found to greatly improve pregnancy rates in women who have a luteal phase defect despite having regular menses and attaining a mature follicle. [4] Progesterone is a white or almost white, crystalline powder or colorless crystals, practically insoluble in water, freely soluble in ethanol, sparingly soluble in acetone and in fatty oils.[5] Progesterone suppositories contains progesterone (200 mg) as an active ingredient and it is a generic drug not manufactured in the Iraqi factories. The aim of study is a necessity to have an a formula for this dosage form with its stability study compatible with specifications of United State pharmacopeia.

Experimental work Several formulations with different suppositories bases were prepared and the selected formula constituents are presented in table (1). The formula contains poly ethylene glycol 600 ( P.E.G), poly ethylene glycol 400 (P.E.G) ,preservative as inactive ingredients. In suitable pyrex beaker transfer the following materials ,P.E.G. 600,P.E.G.400, then heat at 60 ◦C,add the preservative ( methyl paraben and propyl paraben ) to above mixture, mix ( 15) minutes or until clear solution formed, cool to 50 ◦C, then add the stated amount of progesterone to above mixture, mix ( 15) minutes, fill in ( 2) gram plastic molds suppository.

Stability study Stability study was conducted on the product for ( 6 )months at different temperatures ( room temperature(R.T) ( 25 ) ◦C , refrigerator (Ref.) ( 2- 8 )◦C,30◦C) to determine the expiration date and the best storage conditions.

Results and Discussion A torpedo shaped smooth suppository was obtained and the physico – chemical properties are presented in table (1) .On the other hand , stability study is presented in tables (2) and (3). In this study, different formulas of suppostiry were prepared according to the specification of British Pharmacopoeia 2013 and United States pharmacopeia (30) . A torpedo shaped smooth suppository of acceptable consistency was produced and the physicochemical of our formula showed excellent stability at different temperatures (Ref. (2-8) ◦C , R.T.(25)◦C , 30 ◦C). According to this study, the expiration date has been estimated to be not less than (3) years from the date of manufacturing at room temperature ( 25 – 30 ) ◦C . Table -1-The physicochemical changes of progesterone suppository 200 mg with time at different temperatures. Storage time (month) Temp. ◦C % progesteron appearance Zero time R.T 25 103.5 torpedo shaped smooth suppository

1 Ref.(2-8) 103.5 torpedo shaped smooth suppository R.T 25 103.5 torpedo shaped smooth suppository 30 103.39 torpedo shaped smooth suppository

2 Ref.(2-8) 103.39 torpedo shaped smooth suppository R.T 25 103.38 torpedo shaped smooth suppository 30 103.17 torpedo shaped smooth suppository 3

Ref.(2-8) 103.28 torpedo shaped smooth suppository R.T 25 103.13 torpedo shaped smooth suppository 30 102.91 torpedo shaped smooth suppository 4

Ref.(2-8) 103.17 torpedo shaped smooth suppository R.T 25 102.88 torpedo shaped smooth suppository 30 102.66 torpedo shaped smooth suppository 5

Ref.(2-8) 103.06 torpedo shaped smooth suppository R.T 25 102.63 torpedo shaped smooth suppository 30 102.44 torpedo shaped smooth suppository 6

Ref.(2-8) 102.95 torpedo shaped smooth suppository R.T 25 102.38 torpedo shaped smooth suppository 30 102.2 torpedo shaped smooth suppository

Table - 2 -The percentage of Progesterone content suppository in 200 mg drug vaginal suppositories at different temperatures .

Progesterone percentage content Time /day Temperatures ◦C 30 25 2-8 103.39 103.5 103.5 30 103.14 103.38 103.39 60 102.91 103.13 103.28 90 102.66 102.88 103.17 120 102.44 102.63 103.06 150 102.2 102.38 102.95 180 Regarding the expiry date of the prepared formula we followed the Zero – order rate of reaction that is expected for kinetic of progesterone suppository particularly in the first stage of reduction of concentration. T95% =0.1C/K Where: C=concentration at zero time K= rate of reaction T= time

Table -3 -The effect of different temperature on progesterone remain in prepared vaginal suppositories . T95% ( Year) T95% ( day) Temp. ◦C 4 1460 2-8 3.8 1387 25 3.2 1168 30

References 1- Szekeres-Bartho J, Barakonyi I, Polgar B, et al. The role of gamma/delta T cells in progesterone-mediated immunomodulation during pregnancy: a review. Am J Reprod Immunol. 1999;42:44–48. [PubMed]. 2- Meena Khandelwal. Vaginal progesterone in risk reduction of preterm birth in women with short cervix in the midtrimester of pregnancy. Int J Womens Health. 2012; 4: 481–490. [PubMed]

3-Schumacher M, Sitruk-Ware R, De Nicola AF. Progesterone and progestins: neuroprotection and myelin repair. Curr Opin Pharmacol. 2008;8:740–746. [PubMed]. 4- Check JH, Nowroozi K, Wu CH, Adelson HG, Lauer C. Ovulation-inducing drugs versus progesterone therapy for infertility in patients with luteal phase defects. Int J Fertil. 1988;33(4):252–256. [PubMed] 5- British Pharmacopoeia press, 2013, British Pharmacopoeia, U.K.

6- United State Pharmacopoeia press, 2007, U.S.P 30, U.S.A.

الخلاصة نفذ هذا العمل لتطوير منتج دوائي وطني يحتوي على بروجيستيرون 200ملغم على شكل تحاميل مهبلية تستعمل لعلاج العقم عند النساء الغير قادرة على الحمل بسبب نقص هرمون البروجسترون الطبيعي. حضرت عدة تركيبات باستخدام قواعد تحاميل مختلفة (بولي اثيلين كلايكول 400 و 600 واحماض دهنية )، والمواد الحافظة (المثيل بارابين والبروبيل بارابين ) . اختيرت أفضل تركيبة بالاعتماد على الاختبارات الفيزيائية والكيميائية للعينات المحضرة. أجريت دراسة الثباتية على المنتج لمدة ستة أشهر عند درجات حرارة مختلفة (2-8 درجة مئوية، 25 درجة مئوية، 30 درجة مئوية) لتحديد تاريخ انتهاء الصلاحية وأفضل ظروف التخزين. من الدراسة حصلنا على تحاميل مهبلية جيدة التجانس , النعومة والصلابة . حسب تاريخ انتهاء الصلاحية على ان لا يقل عن ثلاث سنوات من تاريخ الإنتاج.