Prodrugs for nitroreductase based cancer therapy-4: Towards prostate cancer targeting: Synthesis of N-heterocyclic nitro prodrugs, Ssap-NtrB enzymatic activation and anticancer evaluation.

In this study, various N-heterocyclic nitro prodrugs (NHN1-16) containing pyrimidine, triazine and piperazine rings were designed and synthesized. The final compounds were identified using FT-IR, 1H NMR, 13C NMR as well as elemental analyses. Enzymatic activities of compounds were conducted by using HPLC analysis to investigate the interaction of substrates with Ssap-NtrB nitroreductase enzyme. MTT assay was performed to evaluate the toxic effect of compounds against Hep3B and PC3 cancer cell lines and healthy HUVEC cell. It was observed that synthesized compounds NHN1-16 exhibited different cytotoxic profiles. Pyrimidine derivative NHN3 and triazine derivative NHN5 can be good drug candidates for prostate cancer with IC50 values of 54.75 µM and 48.9 µM, respectively. Compounds NHN6, NHN10, NHN12, NHN14 and NHN16 were selected as prodrug candidates because of non-toxic properties against three different cell models. The NHN prodrugs and Ssap-NtrB combinations were applied to SRB assay to reveal the prodrug capabilities of these selected compounds. SRB screening results showed that the metabolites of all selected non-toxic compounds showed remarkable cytotoxicity with IC50 values in the range of 1.71-4.72 nM on prostate cancer. Among the tested compounds, especially piperazine derivatives NHN12 and NHN14 showed significant toxic effect with IC50 values of 1.75 nM and 1.79 nM against PC3 cell compared with standart prodrug CB1954 (IC50: 1.71 nM). Novel compounds NHN12 and NHN14 can be considered as promising prodrug candidates for nitroreductase-prodrug based prostate cancer therapy.

Lidocaine and tenoxicam effectiveness for pain relief during Pipelle: Non-randomised double-blind placebo-controlled trial.

OBJECTIVE: To compare the effectiveness of intrauterine lidocaine infusion with lidocaine and intravenous tenoxicam for decreasing the pain levels associated with endometrial biopsy. METHODS: This double-blind, placebo-controlled trial was conducted at Fatih Sultan Mehmet Training and Research Hospital, Istanbul, Turkey, from May to November 2015, and comprised patients undergoing endometrial biopsy with Pipelle. Intrauterine lidocaine infusion, paracervical block with lidocaine, intravenous tenoxicam or 4ml intravenous normal saline administered prior to biopsy. The main outcome measure was pain intensity immediately afterwards and 30minutes after biopsy, determined by a visual analogue scale score. Number Cruncher Statistical System 2007 was used for statistical analyses. RESULTS: Of the 232 participants, intrauterine lidocaine infusion group had 59(25.4%) patients, 57(24.6%) were controls while paracervical block group and intravenous tenoxicam group each had 58(25%) patients. Both visual analogue scale 0 and 30 scores of the control group were significantly higher than the other three groups (p<0.05). Also, the scores of intravenous tenoxicam group were significantly higher than both intrauterine lidocaine infusion and paracervical block with lidocaine groups (p<0.05 each). CONCLUSIONS: Intravenous tenoxicam had a significantly lower effect than intrauterine lidocaine infusion and paracervical block with lidocaine during the early period after the procedure.

Modification of existing antibiotics in the form of precursor prodrugs that can be subsequently activated by nitroreductases of the target pathogen.

The use of existing antibiotics in the form of prodrug followed by activation using enzymes of pathogenic origin could be a useful approach for antimicrobial therapy. To investigate this idea, a common antibiotic, sulfamethoxazole has been redesigned in the form of a prodrug by simple functional group replacement. Upon reductive activation by a type I nitroreductase from a pathogen, the drug displayed enhanced antimicrobial capacity. This strategy could improve the efficacy and selectively of antibiotics and reduce the incidence of resistance.

Iodine status of pregnant women in a metropolitan city which proved to be an iodine-sufficient area. Is mandatory salt iodisation enough for pregnant women?

The objective of this study was to assess the iodine status of pregnant women in a metropolitan city which was stated as iodine sufficient area after salt iodination program. This multicenter, cross-sectional study was carried out on 3543 pregnant women. Age, gestational weeks, smoking, consumption of iodized salt, dietary salt restriction, history of stillbirth, abortus and congenital malformations were questioned. Spot urine samples were analyzed for urine iodine concentration (UIC). The outcomes were: (a) median UIC in three trimesters of pregnancy and (b) frequency of ID among pregnant women. The median UIC was 73 µg/L. The median UIC was 77 µg/L (1-324), 73 µg/L (1-600) and 70 µg/L (1-1650) in three trimesters of pregnancy, respectively (p: 0.14). UIC <50 µg/L was observed in 36.6% (n: 1295) and UIC<150 µg/L was observed in 90.7% (n: 3214) of pregnant women. Only 1% (n: 34) of the pregnant women had UIC levels higher than 500 µg/L. This study showed that more than 90% of the pregnant women in this iodine-sufficient city are facing some degree of iodine deficiency during their pregnancy. A salt iodization program might be satisfactory for the non-pregnant population, but it seems to be insufficient for the pregnant population.

Antiproliferative activity of Humulus lupulus extracts on human hepatoma (Hep3B), colon (HT-29) cancer cells and proteases, tyrosinase, ß-lactamase enzyme inhibition studies.

The aims of this study were to examine the antiproliferation of Humulus lupulus extracts on human hepatoma carcinoma (Hep3B) and human colon carcinoma (HT-29) cell lines along with enzyme inhibitory effects of the crude extracts. Potential cell cytotoxicity of six different H. lupulus extracts were assayed on various cancer cells using MTT assay at 24, 48 and 72 h intervals. Methanol-1 extract has inhibited the cell proliferation with doses of 0.6-1 mg/mL in a time dependent (48 and 72 hours) manner in Hep3B cells with 70% inhibition, while inhibitory effect was not seen in colon cancer cells. Acetone extract has increased the cell proliferation at low doses of 0.1 mg/mL for 72 h in Hep3B cells and 0.1-0.2 mg/mL for 48 and 72 h in HT29 cells. The inhibitory effects of the extracts were compared by relative maximum activity values (V(max)) using proteases such as α-chymotrypsin, trypsin and papain, tyrosinase and ß-lactamase (penicillinase).

Highly stable and reusable immobilized formate dehydrogenases: Promising biocatalysts for in situ regeneration of NADH.

This study aimed to prepare robust immobilized formate dehydrogenase (FDH) preparations which can be used as effective biocatalysts along with functional oxidoreductases, in which in situ regeneration of NADH is required. For this purpose, Candida methylica FDH was covalently immobilized onto Immobead 150 support (FDHI150), Immobead 150 support modified with ethylenediamine and then activated with glutaraldehyde (FDHIGLU), and Immobead 150 support functionalized with aldehyde groups (FDHIALD). The highest immobilization yield and activity yield were obtained as 90% and 132%, respectively when Immobead 150 functionalized with aldehyde groups was used as support. The half-life times (t 1/2) of free FDH, FDHI150, FDHIGLU and FDHIALD were calculated as 10.6, 28.9, 22.4 and 38.5 h, respectively at 35 °C. FDHI150, FDHIGLU and FDHIALD retained 69, 38 and 51% of their initial activities, respectively after 10 reuses. The results show that the FDHI150, FDHIGLU and FDHIALD offer feasible potentials for in situ regeneration of NADH.

A new lipase as a pharmaceutical target for battling infections caused by Staphylococcus aureus: Gene cloning and biochemical characterization.

Staphylococcus aureus lipases along with other cell-wall-associated proteins and enzymes (i.e., catalase, coagulase, protease, hyaluronidase, and ß-lactamase) play important roles in the pathogenesis of S. aureus and are important subject of drug targeting. The appearance of antibiotic-resistant types of pathogenic S. aureus (e.g., methicillin-resistant S. aureus, MRSA) is a worldwide medical problem. In the present work, a novel lipase from a newly isolated MRSA strain from a cow with subclinical mastitis was cloned and biochemically characterized. The mature part of the lipase was expressed in Escherichia coli and purified by nickel affinity chromatography. It displays a high lipase activity at pH 8.0 and 25 °C using p-nitrophenyl palmitate and has a preference for medium-long-chain substrates of p-nitrophenyl esters (pNPC10-C16). Furthermore, in search of inhibitors, the effect of farnesol on the growth of S. aureus and the lipase activity was also studied. Farnesol inhibits the growth of S. aureus and is a mixed-type inhibitor with Ki and Ki (') values of 0.2 and 1.2 mmol L(-1), respectively. A lipase with known properties could not only serve as a template for developing inhibitors for S. aureus but also a valuable addition to enzyme toolbox of biocatalysis. The discovery of this lipase can be potentially important and could provide a new target for pharmaceutical intervention against S. aureus infection.

An unusually cold active nitroreductase for prodrug activations.

A set of PCR primers based on the genome sequence were used to clone a gene encoding a hypothetical nitroreductases (named as Ssap-NtrB) from uropathogenic staphylococcus, Staphylococcus saprophyticus strain ATCC 15305, an oxygen insensitive flavoenzyme. Activity studies of the translation product revealed that the nitroreductase catalyses two electron reduction of a nitroaromatic drug of nitrofurazone (NFZ), cancer prodrugs of CB1954 and SN23862 at optimum temperature of 20 °C together with retaining its maximum activity considerably at 3 °C. The required electrons for such reduction could be supplied by either NADH or NADPH with a small preference for the latter. The gene was engineered for heterologous expression in Escherichia coli, and conditions were found in which the enzyme was produced in a mostly soluble form. The recombinant enzyme was purified to homogeneity and physical, spectral and catalytical properties were determined. The findings lead us to propose that Ssap-NtrB represents a novel nitro reductase with an unusual cold active property, which has not been described previously for prodrug activating enzymes of nitroreductases.

Microneedles in Drug Delivery: Progress and Challenges.

In recent years, an innovative transdermal delivery technology has attracted great interest for its ability to distribute therapeutics and cosmeceuticals for several applications, including vaccines, drugs, and biomolecules for skin-related problems. The advantages of microneedle patch technology have been extensively evaluated in the latest literature; hence, the academic publications in this area are rising exponentially. Like all new technologies, the microneedle patch application has great potential but is not without limitations. In this review, we will discuss the possible limitations by highlighting the areas where a great deal of improvements are required. Emphasising these concerns early on should help scientists and technologists to address the matters in a timely fashion and to use their resources wisely.

Clinical efficacy of dissolvable microneedles armed with anti-melanogenic compounds to counter hyperpigmentation.

BACKGROUND: Hyperpigmentation is a complex physiological process associated with alterations of skin color due to melanin overproduction and distribution. Both intrinsic and extrinsic factors influence discoloration with the involvement of multiple pathways in cooperative manners. Restoring natural skin color requires a multi-targeted approach. Latest advances in anti-melanogenic compounds and microneedle delivery system have presented opportunities for tackling hyperpigmentationsuccessfully. AIMS: This work was aimed at assessing the dermal tolerability and efficacy of hyaluronic acid-based microneedle patches loaded with anti-melanogenic actives for improvements of skin discoloration. PATIENTS/METHODS: The test products consist of hyaluronic acid with niacinamide, ascorbic acid 2-glucoside, tranexamic acid, resveratrol, 4-n-butyl-resorcinol, and Halidrys siliquosa extract. In a monocentric 12 weeks clinical trial, the HA-MNs patches were applied to the affected areas of the faces on subjects with hyperpigmented skin. The color properties of the skin were analyzed spectrophotometrically. RESULTS: The products were tolerated remarkably; none of the subjects informed any primary or cumulative skin responses. Evaluation of color related measurable skin properties provided insight into the general effectiveness: The individual topology measurements showed 51.4% noticeable improvements in hyperpigmented zones, which were also in good agreement with the personal impressions of the subjects determined by questionnaires before and after the treatments. CONCLUSIONS: The concentrated blends of actives in microneedle patches act in a multi-targeted manner, and they could be worked in a complementary fashion for the improvement of skin color and appearance. The study has established the overall convenience of the HA-MNs with the formulation of anti-melogenic compounds against skin discoloration.

Efficacy of bioactive peptides loaded on hyaluronic acid microneedle patches: A monocentric clinical study.

BACKGROUND: Aging skin is a gradual physiological process associated with functional and structural changes of the skin. Both intrinsic and extrinsic factors influence skin aging by the involvement of multiple pathways. Restoring natural skin conditions requires a multi-targeted approach. Recent developments in both bioactive peptides and microneedle delivery system have presented an opportunity for tackling premature skin aging. AIMS: This study was aimed at evaluating the dermal tolerability and efficacy of hyaluronic acid-based microneedle patches loaded with bioactives for restoration of the skin properties including hydration, wrinkle reduction, density, and thickness. PATIENTS/METHODS: The test product of HA-MNs comprises arginine/lysine polypeptide, acetyl octapeptide-3, palmitoyl tripeptide-5, adenosine, and seaweed extracts. In the monocentric 12-week clinical trial, the HA-MNs patches were applied to the outer corner of the right and left eye and a defined area on the volar forearm on healthy subjects with aged skin. Instrumental analysis of skin properties was determined. RESULTS: The product was tolerated excellently; none of the subjects reported any primary or cumulative skin reactions. Assessment of measurable skin properties provides insight into the general effectiveness: The fine lines/wrinkles showed 25.8% noticeable decrease; the skin hydration measurements demonstrated 15.4% improvement; the skin density and thickness in the dermis increased 14.2% and 12.9%, respectively. CONCLUSIONS: The composition of the microneedle patches works in a multi-targeted manner and all ingredients might possibly be acted synergistically for the improvement of skin structure, function, and appearance. The study has demonstrated the overall usefulness of the HA-MNs with careful formulation for skin care applications.

Prodrugs for nitroreductase-based cancer therapy-3: Antitumor activity of the novel dinitroaniline prodrugs/Ssap-NtrB enzyme suicide gene system: Synthesis, in vitro and in silico evaluation in prostate cancer.

Prodrugs for targeted tumor therapies have been extensively studied in recent years due to not only maximising therapeutic effects on tumor cells but also reducing or eliminating serious side effects on healthy cells. This strategy uses prodrugs which are safe for normal cells and form toxic metabolites (drugs) after selective reduction by enzymes in tumor tissues. In this study, prodrug candidates (1-36) containing nitro were designed, synthesized and characterized within the scope of chemical experiments. Drug-likeness properties of prodrug candidates were analyzed using DS 2018 to investigate undesired toxicity effects. In vitro cytotoxic effects of prodrug canditates were performed with MTT assay for human hepatoma cells (Hep3B) and prostate cancer cells (PC3) and human umbilical vein endothelial cells (HUVEC) as healthy control. Non-toxic compounds (3, 5, 7, 10, 12, 15, 17, 19 and 21-23), and also compounds (1, 2, 5, 6, 9, 11, 14, 16, 20 and 24) which had low toxic effects, were selected to examine their suitability as prodrug canditates. The reduction profiles and kinetic studies of prodrug/Ssap-NtrB combinations were performed with biochemical analyses. Then, selected prodrug/Ssap-NtrB combinations were applied to prostate cancer cells to determine toxicity. The results of theoretical, in vitro cytotoxic and biochemical studies suggest 14/Ssap-NtrB, 22/Ssap-NtrB and 24/Ssap-NtrB may be potential prodrug/enzyme combinations for nitroreductase (Ntr)-based prostate cancer therapy.

Biomechanical analysis of spinal implants with different rod diameters under static and fatigue loads: an experimental study.

Spinal implants are commonly used in the treatment of spinal disorders or injuries. However, the biomechanical analyses of them are rarely investigated in terms of both biomechanical and clinical perspectives. Therefore, the main purpose of this study is to investigate the effects of rod diameter on the biomechanical behavior of spinal implants and to make a comparison among them. For this purpose, three spinal implants composed of pedicle screws, setscrews and rods, which were manufactured from Ti6Al4V, with diameters of 5.5 mm, 6 mm and 6.35 mm were used and a bilateral vertebrectomy model was applied to spinal systems. Then, the obtained spinal systems were tested under static tension-compression and fatigue (dynamic compression) conditions. Also, failure analyses were performed to investigate the fatigue behavior of spinal implants. After static tension-compression and fatigue tests, it was found that the yield loads, stiffness values, load carrying capacities and fatigue performances of spinal implants enhanced with increasing spinal rod diameter. In comparison to spinal implants with 5.5 mm rods, the fatigue limits of implants showed 13% and 33% improvements in spinal implants having 6 mm and 6.35 mm rods, respectively. The highest static and fatigue test results were obtained from spinal implants having 6.35 mm rods among the tested implants. Also, it was observed that the increasing yield load and stiffness values caused an increase in the fatigue limits of spinal implants.

PRODRUGS FOR NITROREDUCTASE BASED CANCER THERAPY- 2: Novel amide/Ntr combinations targeting PC3 cancer cells.

The use of nitroreductases (NTR) that catalyze the reduction of nitro compounds by using NAD(P)H in GDEPT (Gene-directed enzyme prodrug therapy) studies which minimize toxicity at healthy cells and increases concentration of drugs at cancer cells is remarkable. Discovery of new prodrug/NTR combinations is necessary to be an alternative to known prodrug candidates such as CB1954, SN23862, PR-104A. For this aim, nitro containing aromatic amides (A1-A23)2 were designed, synthesized, performed in silico ADMET and molecular docking techniques in this study. Prodrug candidates were studied on reduction potentials with Ssap-NtrB by HPLC system. Also, cyototoxic properties and prodrug ability of these amides were investigated using different cancer cell lines such as Hep3B and PC3. As a result of theoretical and biological studies, combinations of A5, A6 and A20 with Ssap-NtrB can be suggested as potential prodrugs/enzyme combinations at NTR based cancer therapy compared with CB1954/NfsB.

DMSO tolerant NAD(P)H recycler enzyme from a pathogenic bacterium, Burkholderia dolosa PC543: effect of N-/C-terminal His Tag extension on protein solubility and activity.

NAD(P)+ dependent formate dehydrogenase (FDH) is an oxidoreductase used as a biocatalyst to regenerate NAD(P)H in reductase-mediated chiral synthesis reactions. Solvent stability and the need to reduce NADP+ to NADPH, due to the high cost of NADPH, are required features in the industrial usage of FDHs. Therefore, we aimed to identify a novel, robust NADP+ dependent FDH and evaluate the effect of N- and C- terminus His tag extensions on protein solubility and activity. Herein, we report a novel, DMSO tolerant formate dehydrogenase (BdFDH), which has dual coenzyme specificity and tolerance to acidic pH, from Burkholderia dolosa PC543. N- and C-terminus His-tagged BdFDHs were expressed separately in Escherichia coli BL21 (DE3). The C-terminal His-tagged BdFDH was soluble and active whereas the N-terminal version was not. The enzyme displays dual coenzyme specificity and resistance to some organic solvents, particularly DMSO, and is able to tolerate acidic pH conditions. The apparent KM values for NADP+, NAD+ and sodium formate (with NADP+), are 1.17, 14.7 and 5.66 mM, respectively. As a result, due to its DMSO tolerance and coenzyme preference, this enzyme can be utilized as an NAD(P)H recycler in several biotransformations particularly when carried out under acidic conditions. Moreover, it can be said that the position of the His tag extension may affect the enzyme solubility and functionality.

Prodrugs for Nitroreductase Based Cancer Therapy- 1: Metabolite Profile, Cell Cytotoxicity and Molecular Modeling Interactions of Nitro Benzamides with Ssap-NtrB.

BACKGROUND: Directed Enzyme Prodrugs Therapy (DEPT) as an alternative method against conventional cancer treatments, in which the non-toxic prodrugs is converted to highly cytotoxic derivative, has attracted an ample attention in recent years for cancer therapy studies. OBJECTIVE: The metabolite profile, cell cytotoxicity and molecular modeling interactions of a series of nitro benzamides with Ssap-NtrB were investigated in this study. METHOD: A series of nitro-substituted benzamide prodrugs (1-4) were synthesized and firstly investigated their enzymatic reduction by Ssap-NtrB (S. saprophyticus Nitroreductase B) using HPLC analysis. Resulting metabolites were analyzed by LC-MS/MS. Molecular docking studies were performed with the aim of investigating the relationship between nitro benzamide structures (prodrugs 1-4) and Ssap-NtrB at the molecular level. Cell viability assay was conducted on two cancer cell lines, hepatoma (Hep3B) and colon (HT-29) cancer models and healthy cell model HUVEC. Upon reduction of benzamide prodrugs by Ssap-NtrB, the corresponding amine effectors were tested in a cell line panel comprising PC-3, Hep3B and HUVEC cells and were compared with the established NTR substrates, CB1954 (an aziridinyl dinitrobenzamide). RESULTS: Cell viability assay resulted in while prodrugs 1, 2 and 3 had no remarkable cytotoxic effects, prodrug 4 showed the differential effect, showing moderate cytotoxicity with Hep3B and HUVEC. The metabolites that obtained from the reduction of nitro benzamide prodrugs (1-4) by Ssap-NtrB, showed differential cytotoxic effects, with none toxic for HUVEC cells, moderate toxic for Hep3B cells, but highly toxic for PC3 cells. CONCLUSION: Amongst all metabolites of prodrugs after Ssap-NtrB reduction, N-(2,4- dinitrophenyl)-4-nitrobenzamide (3) was efficient and toxic in PC3 cells as comparable as CB1954. Kinetic parameters, molecular docking and HPLC results also confirm that prodrug 3 is better for Ssap-NtrB than 1, 2 and 4 or known cancer prodrugs of CB1954 and SN23862, demonstrating that prodrug 3 is an efficient candidate for NTR based cancer therapy.

Biological evaluation and molecular docking studies of nitro benzamide derivatives with respect to in vitro anti-inflammatory activity.

A series of nitro substituted benzamide derivatives were synthesized and evaluated for their potential anti-inflammatory activities in vitro. Firstly, all compounds (1-6) were screened for their inhibitory capacity on LPS induced nitric oxide (NO) production in RAW264.7 macrophages. Compounds 5 and 6 demonstrated significantly high inhibition capacities in a dose-dependent manner with IC50 values of 3.7 and 5.3µM, respectively. These two compounds were also accompanied by no cytotoxicity at the studied concentrations (max 50µM) in macrophages. Molecular docking analysis on iNOS revealed that compounds 5 and 6 bind to the enzyme more efficiently compared to other compounds due to having optimum number of nitro groups, orientations and polarizabilities. In addition, 5 and 6 demonstrated distinct regulatory mechanisms for the expression of the iNOS enzyme at the mRNA and protein levels. Specifically, both suppressed expressions of COX-2, IL-1ß and TNF-α significantly, at 10 and 20µM. However, only compound 6 significantly and considerably decreased LPS-induced secretion of IL-1ß and TNF-α. These results suggest that compound 6 may be a multi-potent promising lead compound for further optimization in structure and as well as for in vivo validation studies.

Probing the substrate specificity of the catalytically self-sufficient cytochrome P450 RhF from a Rhodococcus sp.

Analysis of the substrate specificity of the self-sufficient cytochrome P450 RhF revealed that the enzyme tends to catalyse the dealkylation of substituted alkyl-aryl ethers with shorter alkyl moieties more readily than equivalent compounds with longer alkyl groups.

Neutrophil-to-lymphocyte and platelet-to-lymphocyte ratios: are they useful for predicting gestational diabetes mellitus during pregnancy?

OBJECTIVE: We aimed to investigate whether the neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) could be utilized to screen for gestational diabetes mellitus (GDM). SUBJECTS AND METHODS: NLR and PLR were assessed by retrospective analysis of 762 healthy and pregnant women with GDM. The patients were stratified into four groups, as follows: GDM (n=144), impaired glucose tolerance (n=76), only screen positive (n=238), and control (n=304). RESULTS: The leukocyte, neutrophil, and lymphocyte counts were significantly higher in the study groups compared with the control group (P=0.001; P<0.01). There were no statistically significant differences between the groups with respect to the NLR and PLR (P>0.05). CONCLUSION: We do not recommend that blood NLR and PLR can be used to screen for GDM. However, increase in the leukocyte count is an important marker for GDM as it provides evidence of subclinical inflammation.

A Clinical Experience of Ectopic Pregnancies with Initial Free Intraperitoneal Fluid.

INTRODUCTION: Extra-uterine pregnancy or Ectopic Pregnancy (EP) is a major health problem for pregnant women, presenting as a potentially life-threatening emergency in the first trimester. There are three major options for the treatment of EP: expectant management, surgical treatment and medical management. The presence of free intraperitoneal fluid in EP-diagnosed patients is crucial for treatment planning and evaluation. AIM: To compare the outcomes of both the expectant man-agement and medical treatment with methotrexate (MTX) in ectopic pregnancies with free intraperitoneal fluid. MATERIALS AND METHODS: This retrospective cohort study inclu-ded a total of 91 ectopic pregnancies with or without rupture in which the women had initial free intraperitoneal fluid and were haemodynamically stable. Serial ß-HCG measurements were used to assess the outcome of expectant management and medical treatment with MTX. For the statistical analysis, the SPSS statistical software package, version 22.0 (Chicago, IL, USA), was used. For the quantitative variables that were not distributed normally, the Kruskal-Wallis test and the Mann-Whitney U test were performed for the evaluation of differences between the groups. RESULTS: It was observed that the success rate with expectant management was 81% (initial ß HCG concentration 626±443 mIU/mL). With a single dose of MTX, it was 76% (initial ß HCG concentration 2124±1647 mIU/mL) and with a total single or double dose of MTX, it was 88% (initial ß HCG concentration 2252±78 mIU/mL) from among EP with or without rupture in women with initial free intraperitoneal fluid during diagnosis. There was no significant difference between the groups with regard to ultrasonography findings. CONCLUSION: Expectant management or medical treatment with methotrexate should be the first line treatment for ectopic pregnancies with initial free intraperitoneal fluid, albeit with rupture, in patients who are haemodynamically stable, along with ß-HCG follow-up.