Research Title: Efficacy of Follitropin-A (Gonal-F) Versus Follitropin-B (Puregon) for Controlled Ovarian Stimulation in Women Undergoing in vitro fertilization (IVF)

Author: Yazan Batineh, Published Year: 2019

International Journal of Pharmaceutical Quality Assurance, 10

Faculty: Pharmacy

Abstract: The purpose of this study was to compare the efficacy of two recombinant follicle-stimulating hormone (FSH) on pregnancy rates in infertile patients. Material and Methods: between 2015-2017, 387 females intended to have in vitro fertilization (IVF) for infertility treatment (226 patients use Gonal-F and 161 using Puregon. Results: Serum E2 concentration at hCG time was higher with follitropin-a treated patients, and a larger number of retrieved oocyte result in a large number of the transferred embryo, and high pregnancy rate than Follitropin-b treated patients. Conclusions: Gonal-F (Follitropin-a) is associated with a potential stimulatory effect on ovaries. Puregon (Follitropin-b) was associated with a lower clinical pregnancy rate (PR). E2 level 5-7 days after stimulation can be used as an indicator of the success of IVF.

Keywords: COS, Fertilization, Follitropin-a, Follitropin-b, Gonal-F, IVF, Puregon.

Research Title: Efficacy and Safety of Canagliflozin Compared to Sitagliptin and Glimepride as Add- on Therapy in T2DM

Author: Yazan Batineh, Published Year: 2019

Journal of Global Pharma Technology, 11

Faculty: Pharmacy

Abstract: Diabetes type II is a progressive disease and associated with many complications. Metformin is the firstline pharmacological therapy for type II. However, there is a second anti-diabetic drug can be added for patients who do not achieve sufficient glycemic control with metformin. In this study, we assess the efficacy and safety of newly approved anti-diabetic drugs Canagliflozin compared to Sitagliptin and Glimepiride patient with type II diabetes. Basically, 210 diabetic patients have received Canagliflozin 300 mg or Sitagliptin 100 mg or Glimepiride 4 mg for at least 16 weeks. Previously, the patient's glycemic state was uncontrolled by Metformin monotherapy. Glycated hemoglobin and Fasting blood glucose changes were used as an efficacy assessment from the baseline after metformin discontinuation. The safety profile was determined by comparing patient’s lipid profile, renal function, BUN, renal function and uric acid. Our results had shown a significant reduction in HBA1c and FBS in the three groups after 4 months of treatment (p <0.05). However, there was no significant change in the reducing effect between the three drugs (p= 0.704, 0.521). Canagliflozin and Glimepiride produce more reduction in triglycerides than Sitagliptin through the treatment period, although it is not significant, the change in the effect was significantly higher in Canagliflozin and Glimepiride comparing to Sitagliptin. Unlike Canagliflozin, Glimepiride and Sitagliptin produce a significant reduction in LDL after 4 months of treatment (p = 0.0318, 0.047) and significant difference in the effect (p = 0.042). Canagliflozin cause a significant elevation in LDL after the initiation of therapy (p= 0.034). The effect of Canagliflozin was significantly higher on HDL comparing to Glimepiride and Sitagliptin throughout the treatment (p = 0.048) and as a difference in the effect (p =0.043). Canagliflozin and Glimepiride produce a significant elevation in BUN comparing to Sitagliptin (p = 0. 004, 003), whereas, Canagliflozin and Sitagliptin produce a higher reduction in GFR than Glimepiride. Glimepiride is significantly elevates serum uric acid comparing to Canagliflozin and Sitagliptin with p= 0.0406. Conclusion: Canagliflozin, Sitagliptin, and Glimepiride are effective add-on therapy with metformin for proper control of blood glucose in T2DM. Canagliflozin and Glimepiride improve TGs and LDL greater than Sitagliptin. Sitagliptin produces less elevation in BUN than Canagliflozin and Glimepiride. Canagliflozin and Sitagliptin required renal monitoring throughout treatment

Keywords: T2DM, Canagliflozin, Glimepiride, Sitagliptin

Research Title: Hypolipidemic Efficacy of Artemisia absinthium Extracts in Rabbits

Author: Yazan Batineh, Published Year: 2014

World Applied Sciences Journal, 31

Faculty: Pharmacy

Abstract: The hypolipidemic effects of 70% ethanol extract of Artemisia absinthium in hypercholesterolemic -fed rabbits. Hypercholesterolemia was induced in male rabbits by high cholesterol diet (HCD) (350 mg/kg) for 8 weeks. Hypercholesterolemic rabbits were allocated into groups, treated with simvastatin (SIM 5 mg/kg), different extracts of Artemisia absinthium at two doses of 500, 1000 mg/kg. A normal control group and an HCD control one were used for comparison. During the hall period of the experiment blood samples were collected and serum was analyzed for lipid profile. At the end of the experiment the animals were sacrificed; the heart and the liver were collected and stored at -20°C until assayed. Biochemical analysis of blood serum and tissue (liver and heart muscle) were performed for total cholesterol, total triglycerides, Phospholipid, LDL-C, HDL-C, VLDL, aspartate aminotransferase (AST), alanine aminotransferase (ALT), serum creatine kinase (CK) and total protein. The extract induces a significant decrease in serum cholesterol, triglycerides and CK levels. Blood levels of AST, ALT, triglycerides and total protein unchanged. The tissues lipids profiles of liver and heart muscle showed similar changes in those noticed in serum lipids. The results concluded that Artemisia absinthium ethanolic extract (500, 1000 mg/kg) have potent antihyperlipidemic activity in high cholesterol diet induced hyperlipidemia model and which is equipotent activity when compared with control group.

Keywords: Artemisia absinthium Anti-Hyperlipidemic Activity Triglyceride Cholesterol High Cholesterol Diet Induced Hyperlipidemia

Research Title: Effect of Stevia Consumption on Blood Pressure, Stress Hormone Levels and Anthropometrical Parameters in Healthy Persons

Author: Yazan Batineh, Published Year: 2017

American Journal of Pharmacology and Toxicology, 12

Faculty: Pharmacy

Abstract: Stevia is a natural sweetener containing steviol glycosides known to be several times sweeter than sucrose. It is thought to have several beneficial properties though some evidence state it may have detrimental effects. The aim of this study was to investigate the potential beneficial or harmful effects of stevia consumption by exploring its effects on blood pressure, stress hormone levels and anthropometrical markers in A crossover placebo controlled study was conducted on 16 volunteers randomly assigned to consume either stevia or a placebo (sugar) for one week. The measurements were attained on three different occasions and each volunteer was allowed a 3-day initiation period before baseline and in between interventions. The systolic BP increased following stevia intake from 114.5±12.7 to 119.9±12.9mmHg (p<0.001) and diastolic BP from 70.8±9.4 to 75.7±9.6mmHg (p<0.01). Systolic BP increased slightly after the sugar placebo to 115.3±13.6 mmHg (not significant). The mean free cortisol excreted in urine has increased from 91.8±49.1 to 125.7±60.5nmole/day (p<0.01) after the stevia and to 109.1±42.6nmole/day after the placebo (p = 0.210). The ratio of urinary free cortisol/cortisone showed a statistically significant increase from 1.73±0.78 to 2.65±1.03 after stevia (p<0.0001). Salivary cortisol levels have also increased (p<0.01 at AM) after stevia. Placebo intake did not produce a significant change in salivary cortisol. The ratio of salivary cortisol/cortisone during the stevia has increased only in the morning (from 1.22±0.65 to 1.75±0.72, p = 0.05) and a modest increase in the daily average of salivary cortisol/cortisone. There was small insignificant reduction in weight and BMI after stevia intervention (p = 0.246 and p = 0.249 respectively). In conclusion, we have shown that short term stevia intake produced a small but significant increase in BP and effect on body weight and BMI were not significant. The rise in BP might be due to the increase in cortisol levels and cortisol/cortisone ratio indicating that stevia may possibly inhibit 11β-HSD2 enzyme by reducing the conversion of cortisol into cortisone. Therefore caution should be taken by the public who want to consume stevia for longer period of time as a weight reducing sweetener.

Keywords: Stevia, Sweeteners, Blood Pressure, BMI, Glucocorticoids, 11β-HSD

Research Title: Formulation and evaluation of diclofenac controlled release matrix tablets made of HPMC and Poloxamer 188 polymer: An assessment on mechanism of drug release.

Author: Yazan Batineh, Published Year: 2018

Pak J Pharm Sci., 31

Faculty: Pharmacy

Abstract: In this study, hydrophilic hydroxypropyl methylcellulose matrices with various concentrations of Poloxamer 188 were used in the development of oral controlled release tablets containing diclofenac sodium. Four formulations of hydrophilic matrix tablets containing 16.7% w/w HPMC and 0, 6.7, 16.7 and 25.0% w/w Poloxamer 188, respectively, were developed. Tablets were prepared by direct compression and characterized for diameter, hardness, thickness, weight and uniformity of content. The influence of various blends of hydroxypropyl methylcellulose and Poloxamer 188 on the in vitro dissolution profile and mechanism of drug release of was investigated. In the four formulations, the rate of drug release decreased with increasing the concentration of Poloxamer 188 at the initial dissolution stages due to the increase in the apparent viscosity of the gel diffusion layer. However, in the late dissolution stages, the rate of drug release increased with increasing Poloxamer 188 concentration due to the increase in wettability and dissolution of the matrix. The kinetic of drug release from the tablets followed non-Fickian mechanism, as predicted by Korsmeyer-Peppas model, which involves diffusion through the gel layer and erosion of the matrix system.

Keywords: Poloxamer 188, iclofenac sodium, hydroxypropyl methylcellulose, dissolution rate, controlled release.

Research Title: A NOVEL BRAZILIAN SPIDER TOXIN ANALOGUE IS ANTINOCICEPTIVE ACTING VIA THE CANNABINOID SYSTEM

Author: Wafa Moh'd Khair Hourani, Published Year: 2018

The 28th Annual International Cannabinoid Research Society Symposium on the Cannabinoids, Leiden university, the Netherlands

Faculty: Pharmacy

Abstract: Peptide toxins produced by a variety of organisms have evolved with different targets, including GPCR, enzymes and ion channels, where there are many examples of ligands with high potency and selectivity. The Brazilian scientific team developed peptide analogues of an active component of the venom of the Brazilian wandering spider (Phoneutria nigriventer). The synthetic nontoxic peptide PnPP-19 (BR 10 2012 020800-8 A2) induced antinociception involving the inhibition of neutral endopeptidase and activation of CB1 cannabinoid receptors as well as µ and δ opioid receptors (Freitas, Br J Pharmacol PMID: 26947933). In this work, a novel synthetic nontoxic peptide, Pn1 (11 amino acid residues, MW= 1.35 kDa, in patent application) modeled from δCtenitoxin-Pn1a toxin was examined. Nociceptive thresholds were measured by the in vivo paw pressure test on male Wistar rats (180- 200 g). First, to investigate the role of Pn1 in nociception, the peptide was injected (1.5, 2.5, 5 and 10 µg per paw) or vehicle (50 µl) into rat paws-that were hyperalgesic following the administration of carrageenan (250 µg). Pn1 induced a dose-dependent antinociceptive response, with the maximal effects at 10 µg per paw, which peaked at 30 min post-administration. The role of the CB1 cannabinoid receptor in these effects was investigated with intraplantar co-administration of AM251 (40, 80 and 160 µg per paw). The highest dose totally inhibited the antinociceptive effect of Pn1 (10 µg per paw). Moreover, we investigated whether enzymatic degradation by FAAH, MAGL and anandamide endocannabinoid transporter pathways could be contributing to the antinociceptive activity using intraplantar administration of JZL184 (14 µg per paw); MAFP, (4 µg per paw); or VDM11 (40 µg per paw). The results showed that inhibition of MAGL or the transporter potentiated the antinociceptive activity of the Pn-1. In in vitro studies, we evaluated the selectivity of Pn1 using CHO-CB1 and CHO-CB2 human receptors and [35S]-GTPγS binding assays. The preliminary results indicated that Pn1 (10 µM) appears to exhibit selectivity and affinity for the CB1 receptor. Further studies of the downstream signalling pathways for the CB1 receptor (calcium elevation and ERK1/2 phosphorylation), however, failed to show significant effects of Pn1 (10 µM). Antinociception induced by Pn1 appears to involve the inhibition of MAGL and anandamide endocannabinoid transporter and the selective activation of CB1 receptors. This novel peptide could be useful as a new antinociceptive drug candidate

Keywords: cannabinoid receptors,Peptide toxins, Antinociception

Research Title: COUPLING OF THE HUMAN RECOMBINANT CB2 CANNABINOID RECEPTOR TO AKT/PROTEIN KINASE B ACTIVATION/PHOSPHORYLATION IN CHINESE HAMSTER OVARY CELLS

Author: Wafa Moh'd Khair Hourani, Published Year: 2018

The 28th Annual International Cannabinoid Research Society Symposium on the Cannabinoids, Leiden university, the Netherlands

Faculty: Pharmacy

Abstract: Background: Cannabinoids affect the proliferation and differentiation of neural precursor cells in rats and mice. In oligodendrocytes, cannabinoid receptors influence stem cell survival and differentiation via the phosphatidylinositol 3-kinase:protein kinase B (PI3K-Akt, http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=285) signalling pathway(Gomez, Sanchez-Rodriguez et al. 2011). Recruitment to the plasma membrane and binding to phosphatidylinositol 3,4,5-trisphosphate allows phosphorylation and activation of Akt. The main objective of this investigation was to quantify the coupling of CB2 receptors to activation of the Akt signalling pathway. Methods: Chinese hamster ovary cells were stably transfected with human recombinant CB2 cannabinoid receptors. Activation of the Akt signalling pathway was assessed using a phosphopan-Akt antibody and in cell immunocytochemistry in 96-well microtitre plates quantified with a Licor Odyssey using fetal bovine serum as a positive control. Results: Multiple CB2 cannabinoid receptor agonists were investigated including the endogenous ligands; 2-AG and anandamide, as well as the synthetic ligands; CP55940, fenofibrate, HU210 ,WIN55,212-2, JWH015 and JWH133. A rank order of potency was CP55490 (pEC50 7.9 ± 0.04) > WIN55,212-2 (7.6 ± 0.1), HU210 (7.4 ± 0.1) > fenofibrate (7.1 ± 0.1), JWH015 (7.0 ± 0.1) > 2- AG (6.7 ± 0.2), JWH133 (6.3 ± 0.1) > anandamide (5.8 ± 0.1). The maximal effect induced by the different agonists was indicated as the Emax relative to the FBS response with 2-AG producing a maximum response at 149 ± 4 %, HU210 (142 ± 10), CP55940 (139 ± 12), fenofibrate (139 ± 11), WIN55,212-2 (116 ± 10), JWH015 (100 ± 10), JWH133 (98 ± 7), anandamide (71 ± 13). The phosphorylation/activation of the Akt signalling pathway was significantly attenuated by pretreatment with the CB2 selective antagonist AM630 (1 µM) or pertussis toxin (100 ng/mL). Conclusion This is the first study to quantify coupling of the CB2 receptor to Akt phosphorylation using cell immunocytochemistry. In common with other signalling pathways, 2-AG appears to act as a full agonist, while anandamide is a partial agonist, in the phosphorylation of Akt.

Keywords: Cannabinoids, cannabinoid receptors,anandamide, protein kinase B

Research Title: Coupling of the human recombinant CB2 cannabinoid receptor to extracellular signal-regulated kinase and calcium elevation in Chinese hamster ovary cells

Author: Wafa Moh'd Khair Hourani, Published Year: 2017

8th European Workshop on Cannabinoid Research, University of Roehampton, London, UK

Faculty: Pharmacy

Abstract: Background: CB1 cannabinoid receptors are characterised as coupling via the Gi/o family of G proteins to the inhibition of adenylyl cyclase activity and the activation of extracellular signal-regulated kinase (ERK)1. CB1 receptors in neurones also inhibit voltage-gated calcium channels, activate potassium channels and stimulate calcium mobilisation1. CB2 cannabinoid receptors are described to inhibit adenylyl cyclase and activate ERK, but not to regulate ion channel function1. In this study, we have investigated the coupling of human recombinant CB2 cannabinoid receptors expressed in Chinese hamster ovary cells. Methods: Human recombinant CB2 cannabinoid receptors were expressed in Chinese hamster ovary cells. ERK activation was assessed using in cell immunocytochemistry quantified with a Li-Cor Odyssey, while intracellular calcium levels were assessed using fluo-4 with a FlexStation with ATP as a positive control. Results:A range of cannabinoid agonists both endogenous (anandamide and 2-arachidonoylglycerol), natural (THC, β-caryophyllene and caryophyllene N-oxide) and synthetic (CP55940, WIN55,212-2, fenofibrate,HU210, JWH133, and JWH015) were tested. The following agents evoked maximal responses similar to the ATP (78-90 %) with the rank order: WIN55212-2 (pEC50 7.7 ± 0.1)> HU210 (7.4 ± 0.2)> fenofibrate (7.2 ± 0.1), JWH015 (7.2 ± 0.1)>2-AG (6.8 ± 0.1)>JWH133 (6.5 ± 0.1). AEA evoked a reduced maximal response (51%, 5.7 ± 0.2), while β-caryophyllene and caryophyllene N-oxide were inactive. 1 µM AM630, a CB2-selective antagonist, or pretreatment with 100 ng/mL pertussis toxin blocked responses to all these agonists. Calcium ion levels were stimulated by cannabinoid agonists, albeit in a slower manner than ATP. These responses were quantified as area under the curve: WIN55212-2 (21 ± 5 % ATP response, 7.3 ± 0.1) , HU210 (24 ± 3, 6.8 ± 0.3), CP55940 (37 ± 5, 6.7 ± 0.3), JWH015 (24 ± 3, 6.4 ± 0.1), fenofibrate (26 ± 3, 6.3 ± 0.2), 2-AG (34 ± 6, 6.2 ± 0.2), JWH133 (29 ± 7, 6.0 ± 0.1) , AEA (11 ± 1, 5.7 ± 0.3). Responses to THC failed to reach statistical significance, while those to β-caryophyllene and caryophyllene N-oxide were not distinguishable from background. Calcium responses to cannabinoid ligands were blocked by pretreatment with 1 µM AM630 or 100 ng/mL pertussis toxin. Cannabinoid ligand-evoked calcium responses were unaltered in the presence of 10 µM U0126, a selective inhibitor of ERK activation. Conclusions: This is the first study to determine coupling of the CB2 cannabinoid receptor to calcium elevation in recombinant expression.

Keywords: cannabinoid receptors,extracellular signal-regulated kinase,calcium

Research Title: A Voltammetric Sensor Based on Iodine-Coated Platinum Electrode for Determination of Iron in Blood Serum

Author: Wafa Moh'd Khair Hourani, Published Year: 2018

ANALYTICAL & BIOANALYTICAL ELECTROCHEMISTRY , Volume 10 , Number 1

Faculty: Pharmacy

Abstract: A novel, simple, convenient, inexpensive and green voltammetric sensing for iron in human blood serum was developed. The method is based on linear sweep voltammetry at an iodine-coated polycrystalline platinum electrode. A miniaturized home-made small scale 0. 5-mL cell was used for the analysis. Oxidation of iron at the iodine-coated platinum electrode was manifested by a peak centered at 0. 5 V. The established calibration curve for the relationship between iron (II) concentration and current extracted from the linear sweep voltammograms revealed an excellent linearity(R2=0. 9923). The calculated lowest detection limit, LOD, is 0. 26 ppm and the limit of quantification is 0. 87 ppm. Recovery studies for a concentration of 1. 0 ppm yielded a value of 1. 02 ppm (102% percent recovery). No interference with other common essential elements in blood serum was observed which attests to the suitability and accuracy of the method. The developed method was applied to analysis of real blood samples. The results of analysis were compared with the results from conventional standard methods used in medical laboratories where an excellent agreement between the two analytical methods was observed.

Keywords: Voltammetric sensors,Iodine-coated platinum electrode,Serium analysis,Iron determination in blood

Research Title: n−3 polyunsaturated N -acylethanolamines are CB 2 cannabinoid receptor-preferring endocannabinoids

Author: Wafa Moh'd Khair Hourani, Published Year: 2018

Molecular and Cell Biology of Lipids , Volume 1863, Issue 1

Faculty: Pharmacy

Abstract: Anandamide, the first identified endogenous cannabinoid and TRPV1 agonist, is one of a series of endogenous N-acylethanolamines, NAEs. We have generated novel assays to quantify the levels of multiple NAEs in biological tissues and their rates of hydrolysis through fatty acid amide hydrolase. This range of NAEs was also tested in rapid response assays of CB1, CB2 cannabinoid and TRPV1 receptors. The data indicate that PEA, SEA and OEA are not endocannabinoids or endovanilloids, and that the higher endogenous levels of these metabolites compared to polyunsaturated analogues are a correlate of their slow rates of hydrolysis. The n−6 NAEs (AEA, docosatetraenoyl and docosapentaenoyl derivatives) activated both CB1 and CB2 receptors, as well as TRPV1 channels, suggesting them to be ‘genuine’ endocannabinoids and ‘endovanilloids’. The n−3 NAEs (eicosapentaenoyl, docosapentaenoyl and docosahexaenoyl derivatives) activated CB2 receptors and some n−3 NAEs (docosapentaenoyl and docosahexaenoyl derivatives) also activated TRPV1 channels, but failed to activate the CB1 receptor. We hypothesise that the preferential activation of CB2 receptors by n−3 PUFA NAEs contributes, at least in some part, to their broad anti-inflammatory profile.

Keywords: AnandamideEndocannabinoidsCannabinoid receptorsVanilloid receptorsFatty acid amide hydrolasePolyunsaturated fatty acids