A critical assessment on stability behaviour of Vorinostat using LC-MS-QTOF with H/D exchange and NMR.

Vorinostat is the first USFDA-approved HDAC inhibitor for the treatment of cutaneous t-cell lymphoma. Vorinostat was exposed to ICH-recommended hydrolytic (acid, base, and neutral), oxidative, thermal, and photolytic stress conditions to understand the degradation behaviour. A Stability indicating LC method was developed and validated for separating and identifying forced degradation products. Under different stress conditions, six degradants were identified and characterized by LC-HRMS, MS/MS, and hydrogen-deuterium exchange mass studies. Vorinostat was found to be highly susceptible to the acidic and basic environment. In contrast, the drug substance was stable in the solid state under thermal and photolytic conditions whereas, it was found moderately stable when photolytic stress was provided to dissolved state of Vorinostat in acetonitrile-water. The degradants were identified as 7-amino-N-phenylheptanamide, 8-hydrazineyl-8-oxo-N-phenyloctanamide, 8-oxo-8-(phenylamino)octanoic acid, 8-oxo-8-(2-(7-oxo-7-(phenylamino)heptyl)hydrazineyl)-N-phenyloctanamide, 8,8'-(1-hydroxyhydrazine-1,2-diyl)bis(8-oxo-N-phenyloctanamide), and N1-((8-oxo-8-(phenylamino)octanoyl)oxy)-N8-phenyloctanediamide. The mechanistic explanation for the formation of each degradant in stability conditions has also been derived. The major degradants were also isolated/synthesized and characterized through 1H NMR for preparing impurity standards. Additionally, in-silico toxicity of the degradants was predicted in comparison to the drug, to identify whether any degradant has any specific type of toxicity and requires special focus to set specification limits during formulation development. The predicted toxicity indicated that the degradants have similar safety profile as that of the drug and specification can be set as per general impurity guideline.

Drug-Excipient Compatibility Study Through a Novel Vial-in-Vial Experimental Setup: A Benchmark Study.

Drug-excipient compatibility study (DECS) is one of the critical steps during pre-formulation studies to select the appropriate excipient to obtain a stable formulation/dosage form. As such, there is no recommended guideline for DECS. Further, the previously reported studies and protocols followed by various pharmaceutical industries are very lengthy and laborious. Therefore, to improve the existing study strategies and rapid screening of suitable excipients during formulation development, a novel vial-in-vial approach has been proposed. The devised approach was compared with the previously reported conventional approaches using six different drugs with multiple marketed formulations from different manufacturers for each drug. To validate the proposed novel approach, several reported strategies/methodologies have been executed such as exposure of formulations with and without primary packaging, crushed blend with and without water, and/or acetonitrile at accelerated stability condition of 40°C/75% RH for 3 to 6 months and compared with the novel approach. Eventually, all the samples were subjected to HPLC analysis to evaluate the degradation behaviour. The results suggested that the novel approach demonstrated discriminating results with significant degradation as compared to the conventional approaches. Consequently, exercising this methodology for screening the excipients is expected to shorten the drug development cycle by many folds. Moreover, it has also been anticipated that the developed novel approach would prevent the occurrence of late-stage surprises during stability studies.

Modified NAP test: A simple and Responsive Nitrosating Methodology for Risk Evaluation of NDSRIs.

N-Nitroso compounds have been listed as one of the cohorts of concern as per ICH M7. In recent years, the regulatory focus has shifted from common nitrosamines to nitroso-impurities of drug products. Thus, the detection and quantification of unacceptable levels of nitrosamine drug substance-related impurities are of great concern for analytical scientists during drug development. Moreover, risk assessment of nitrosamines is also an essential part of the regulatory filling. For risk assessment, the Nitrosation Assay Procedure suggested by WHO expert group in 1978 is being followed. However, it could not be adopted by the pharmaceutical industries due to the limitation of drug solubility and artefact formation in the test conditions. In this work, we have optimized an alternative nitrosation test to investigate the likelihood of direct nitrosation. The technique is simple, where the drug solubilized in an organic solvent is incubated at 37°C with a nitrosating agent named tertiary butyl nitrite in a 1:10 molar ratio. LC-UV/MS-based chromatographic method was developed to separate drug substances and respective nitrosamine impurities using the C18 analytical column. The methodology was successfully tested on five drugs with varying structural chemistry. The procedure is straightforward, effective, and quick for the nitrosation of secondary amines. This modified nitrosation test and WHO prescribed nitrosation test have been compared and found that the modified methodology is more effective and time-saving.

Hyphenated liquid chromatography - diode array detection - charged aerosol detection - high resolution - multistage mass spectrometry with online hydrogen/deuterium exchange: One stop solution for pharmaceutical impurity profiling.

Hyphenation of different analytical techniques has always been advantageous in structural characterization as it saves time, money and resources. In the pharmaceutical sector, chromatography-based impurity profiling, including identification, characterization, and quantification in drug substances or finished products, is of utmost importance to comply with quality, patient safety and regulatory requirements. These impurities are monitored using LC-UV/DAD and identified and/or characterized using HRMS and MS/MS. LC analysis usually yields the area percent purity of the targeted peak, however, this is not sufficient for pharmaceutical purposes; where the regulatory requirement is to report impurities in percent weight by weight. Unfortunately, the non-availability of impurity standards and relative response factors at an early stage of drug development, risks the product quality due to the inability of the method to differentiate percent purity, and percent weight by weight. Hence, there is a need for a distinctive way of determining the relative response factor. In the current study, a unique hyphenation has been employed by integrating LC with DAD, CAD, and HRMSn with hydrogen-deuterium exchange. The LC flow, post-DAD detection has been diverted to CAD with an inverse gradient for relative response factor determination and MS Orbitrap for exact mass, and MSn fragmentation. A separate infusion pump has been incorporated to infuse D2O on a need basis, which can perform partial hydrogen deuterium exchange for determining the number of labile hydrogens in the impurity structure. This hyphenation has been validated with four model compounds and a total of nineteen chromatographic peaks. The technique provides ample information for their qualitative analysis along with percent weight-by-weight values, which fulfils the regulatory requirements and can be used as one-stop solution for impurity profiling.

Quantification of Anti-Osteoporotic Anabolic Peptide in Stealth Lipid Nanovesicles Through Validated RP-HPLC Method.

BACKGROUND: Teriparatide is a recombinant fragment of human parathyroid hormone, a potent osteoanabolic agent used for osteoporosis. OBJECTIVE: The present study endeavored to develop a simple, rapid, and reliable reverse phase-high performance liquid chromatography (RP-HPLC) method for the determination of teriparatide in pegylated lipid nanovesicles for rapid formulation development/optimization. METHOD: A rapid RP-HPLC-based analytical method was developed for the quantification of teriparatide in pegylated lipid nanovesicles. The method was optimized on a Waters XBridge C18 (4.6 × 150 mm, 10 µm) column with a mobile phase consisting of 0.1% formic acid in water and acetonitrile both in a linear gradient program. In the method, a short run time of 9 min was achieved at a flow rate of 1.0 mL/min with an injection volume of 50 µL at a detection wavelength of 210 nm. The developed method was validated according to the ICH Q2 (R2) guideline. The method was applied for the quantification of teriparatide in prepared pegylated lipid nanovesicles. Teriparatide encapsulated pegylated lipid nanovesicles were prepared by the ethanol injection method. Further, these vesicles were characterized for % entrapment efficiency (%EE), particle size, zeta potential, and morphology by Cryo-SEM. RESULTS: The teriparatide was eluting at 4.8 min in the run. Further, for the method validation, the linear relationship between concentration and response was established over the concentration range of 50-250 µg/mL with the R2 > 0.999. The method sensitivity was shown with LOD and LOQ with the value of 100 ng/mL and 500 ng/mL, respectively. The method was found to be accurate and precise with the recovery ranging in 100 ± 2% and RSD <2%, respectively. Minor deliberate changes proved the robustness of the developed method. CONCLUSIONS: These results indicated that the developed and validated method is accurate, precise, rapid, reliable, and fit for the quantification of teriparatide in different formulations. HIGHLIGHTS: The RP-HPLC method was developed and validated for the quantification of teriparatide from novel pegylated lipid nanovesicles.

Carbamazepine mitigates parenteral nutrition-associated liver disease in a novel ambulatory piglet model.

BACKGROUND: Parenteral nutrition (PN) remains a critical therapeutic option in patients who cannot tolerate enteral feeding. However, although lifesaving, PN is associated with significant side effects, including liver injury, the etiology of which is multifactorial. Carbamazepine (CBZ), an antiepileptic medication, is known to modulate hepatic fibrosis and hepatocellular injury in a variety of liver diseases. We hypothesized that CBZ could prevent PN-associated liver disease (PNALD), which we tested by using our novel ambulatory PN piglet model. METHODS: Piglets were fitted with jugular catheters and infusion pumps for PN and randomized to enteral nutrition (n = 7), PN (n = 6), or PN with parenteral CBZ (n = 6) for 2 weeks. Serum and liver tissue were analyzed via light microscopy, quantification of serum liver injury markers, Ki67 and cytokeratin-7 indexing, and real-time quantitative polymerase chain reaction. RESULTS: PN-fed piglets in our model developed manifestations of PNALD-particularly, increased serum bilirubin, gamma-glutamyltransferase, liver cholestasis, and Ki67 expression compared with that of EN-fed animals (P < 0.03). CBZ therapy in PN-fed animals led to a significant reduction in these markers of injury (P < 0.05). Investigation into the mechanism of these therapeutic effects revealed increased expression of sterol regulatory element-binding protein 1 (SREBP-1), peroxisome proliferator-activated receptor alpha (PPAR-α), and fatty acid binding protein (FABP) in PN-fed animals receiving CBZ (P < 0.03). Further investigation revealed increased LC3 expression and decreased lysosomal-associated membrane protein (LAMP1) expression with CBZ (P < 0.03). CONCLUSION: CBZ administration mitigates PNALD severity, suggesting a novel therapeutic strategy targeting PN-associated side effects, and may present a paradigm change to current treatment options.

Effectiveness of social distancing interventions in containing COVID-19 incidence: International evidence using Kalman filter.

The epidemiological literature has widely documented the importance of social distancing interventions in containing the spread of the COVID-19 pandemic. However, the epidemiological measure of virus reproduction, R0, provides a myopic view of containment, especially when the absolute number of cases is still high. The paper investigates cross-country variations concerning the impact of social distancing interventions on COVID-19 incidence by employing a statistical measure of containment, which models the daily number of cases as a structural time-series, state-space vector. Countries that adopt strict lockdown policies and provide economic support in the form of income augmentations and debt relief improve the response towards the pandemic. Countries like China and South Korea have been most influential in containing the spread of infections. European nations of France, Italy, Spain and the UK are witnessing a second wave of the virus, indicating that re-opening the European economy perhaps has instigated an exponential spread.

Assessment of Bone Quality Using Radiogrammetric Parameters of Proximal Humerus in India: Defining the Osteoporotic Fracture Risk Limit Value and its Reliability.

BACKGROUND: World health organization (WHO) has defined osteoporosis clinically on the basis of bone mineral density (BMD) measurement by dual-energy X-ray absorptiometry (DXA) scan and the presence of fractures. This facility is expensive and not readily available in majority of the centers in India. The authors have attempted to study defined measurements on radiographs (radiogrammetric parameters) to diagnose osteoporosis in Indian population. PATIENTS AND METHODS: We prospectively studied 200 proximal humerus radiographs for measuring radiogrammetric parameters and divided into Group A and B (N = 100 in each group). Group A involved patients with age < 50 years and without any illness affecting bone quality. Group B involved patients with age > 50 years and sustained acute osteoporotic fractures of distal radius/ anterior wedge vertebral fracture/intertrochanteric fracture following trivial trauma. Three parameters (cortical thickness, cortical index and deltoid tuberosity index) were measured by 3 observers at 2 different occasions. RESULTS: The mean age of patients was 37.87 years and 58.38 years for group A and B, respectively. The 'cortical thickness' of the proximal humerus diaphysis had the mean value for Group A and B to be 0.4 ± 0.07 cm and 0.33 ± 0.06 cm respectively. The mean values for the 'cortical index' of proximal humerus came out to be 0.4 ± 0.07 for group A and 0.32 ± 0.06 for group B. The 'deltoid tuberosity index' measurements showed the mean values for group A and B were 1.81 ± 0.23 and 1.55 ± 0.16, respectively. Inter-observer reliability for single measures was excellent for deltoid tuberosity index (ICC 0.8077) and good for cortical thickness (ICC 0.7032) and cortical index (ICC 0.7357). Observer 1 had excellent intra-observer reliabilities for all the three parameters. Observer 2 and 3 had excellent reliability for deltoid tuberosity index and good intra-observer reliability for cortical thickness and cortical index. The cortical thickness had a cut off of ≤ 0.372 cm with a sensitivity of 86.02 and specificity of 82.12. The cortical index had a cut off of ≤ 0.378 with a sensitivity of 89.16 and specificity of 84.22. The deltoid tuberosity index had a cut off of ≤ 1.684 with a sensitivity of 96.61 and specificity of 84.08. CONCLUSION: The outcome of this study is likely to help in early diagnosis of osteoporosis at the community level in the absence of DXA scan as it identifies threshold values for radiogrammetric parameters which can be a predictor of the osteoporosis. The deltoid tuberosity index was found to be the most suitable of these parameters.

Role of Bile Acids and Gut Microbiota in Parenteral Nutrition Associated Injury.

Total Parenteral Nutrition (TPN) is a life-saving therapy where all nutritional requirements are provided intravenously. While this therapy is essential for individuals unable to process their nutritional needs enterically, significant complications arise such as intestinal failure associated liver injury (IFALD). IFALD includes hepatic steatosis, cholestasis, inflammation, ultimately progressing to cirrhosis and portal hypertension and some patients may need liver transplantation. The exact mechanism underlying this condition is not well understood, but studies have recently suggested that changes in gut microbiota and intraluminal bile acid signaling are known to play a role in the development of IFALD. In enterohepatic circulation with normal enteral nutrition, gut Farnesoid X Receptor (FXR) is activated by bile acids, which triggers the release of Fibroblast Growth Factor 19 (FGF19) into portal circulation. FGF19 serves to regulate intrahepatic bile acid synthesis with enteric nutrition. This signaling pathway is impaired in TPN as studies indicate decreased serum levels of FGF19 in subjects receiving TPN. Finally, gut microbiota is severely altered in TPN due to intestinal hypomobility. The shift in gut microbiota affects our immune response and promotes endotoxins that negatively affect liver function. Targeting the pathways affecting gut microbiota and bile acid signaling has promise in treating TPN associated injuries.

Target specific intracellular quantification of etoposide by quadrupole-time of flight based mass spectrometric method.

Etoposide (ETP), a widely used chemotherapeutic agent has an intracellular target site of action. Unfortunately, the concentration of ETP in plasma does not properly reflect the concentration in its intracellular site of action. As per our knowledge, no reported bioanalytical method is available for intracellular quantification of ETP. In this research, we developed an LC-MS/MS method to quantitate ETP in intracellular compartments of MCF-7 cells. The Abcam nuclear extraction kit was used for extracting the nuclear and cytosolic protein from MCF-7 cells. The method showed excellent linearity in the 20-1000 ng/mL range. The intra and inter-day precision (%CV) including LLOQ were found to be in the range of 2.19-16.96% and 6.71-11.21%, respectively, with an accuracy of 86.87 to 110.37% and 93.03 to 100.50%, respectively. The concentration of ETP in nuclear and cytosolic fraction was successfully quantitated using the developed method. The developed method can be applied to understand the efficacy of different formulations based on the intracellular ETP concentration in vitro. It can be considered as a model method for quantification of other similar categories of drugs in their actual intracellular site of action after required optimization in the methodology.

Impaired Gut-Systemic Signaling Drives Total Parenteral Nutrition-Associated Injury.

BACKGROUND: Total parenteral nutrition (TPN) provides all nutritional needs intravenously. Although lifesaving, enthusiasm is significantly tempered due to side effects of liver and gut injury, as well as lack of mechanistic understanding into drivers of TPN injury. We hypothesized that the state of luminal nutritional deprivation with TPN drives alterations in gut-systemic signaling, contributing to injury, and tested this hypothesis using our ambulatory TPN model. METHODS: A total of 16 one-week-old piglets were allocated randomly to TPN (n = 8) or enteral nutrition (EN, n = 8) for 3 weeks. Liver, gut, and serum were analyzed. All tests were two-sided, with a significance level of 0.05. RESULTS: TPN resulted in significant hyperbilirubinemia and cholestatic liver injury, p = 0.034. Hepatic inflammation (cluster of differentiation 3 (CD3) immunohistochemistry) was higher with TPN (p = 0.021). No significant differences in alanine aminotransferase (ALT) or bile ductular proliferation were noted. TPN resulted in reduction of muscularis mucosa thickness and marked gut atrophy. Median and interquartile range for gut mass was 0.46 (0.30-0.58) g/cm in EN, and 0.19 (0.11-0.29) g/cm in TPN (p = 0.024). Key gut-systemic signaling regulators, liver farnesoid X receptor (FXR; p = 0.021), liver constitutive androstane receptor (CAR; p = 0.014), gut FXR (p = 0.028), G-coupled bile acid receptor (TGR5) (p = 0.003), epidermal growth factor (EGF; p = 0.016), organic anion transporter (OAT; p = 0.028), Mitogen-activated protein kinases-1 (MAPK1) (p = 0.037), and sodium uptake transporter sodium glucose-linked transporter (SGLT-1; p = 0.010) were significantly downregulated in TPN animals, whereas liver cholesterol 7 alpha-hydroxylase (CyP7A1) was substantially higher with TPN (p = 0.011). CONCLUSION: We report significant alterations in key hepatobiliary receptors driving gut-systemic signaling in a TPN piglet model. This presents a major advancement to our understanding of TPN-associated injury and suggests opportunities for strategic targeting of the gut-systemic axis, specifically, FXR, TGR5, and EGF in developing ameliorative strategies.

Development and validation of an ambulatory piglet model for short bowel syndrome with ileo-colonic anastomosis.

IMPACT STATEMENT: Short bowel syndrome is associated with significant comorbidities and mortality. This study is important as unlike current systems, it provides a validated piglet model which mirrors anatomical, histological, and serological characteristics observed in human SBS. This model can be used to advance knowledge into mechanistic pathways and therapeutic modalities to improve outcomes for SBS patients. This study is novel in that in addition to significant reduction in the remnant bowel and noted liver disease, we also developed a method to emulate ileocecal valve resection and described gut adaptive responses which has important clinical implications in humans.

Is age just a number: Exploring fear, anxiety, and coping in individuals during COVID-19.

INTRODUCTION: The current pandemic coronavirus (COVID-19) is a novel disease with no standardized course of treatment and cure. This novelty, origin, and dispersion of COVID-19 have been reported to cause chaos, anxiety, and fear among the general population. AIM: The present study aimed to explore the correlation between different age groups, in relation to their general coping strategies, fear, and anxiety due to COVID-19. METHODOLOGY: The research study employed a total sample of 135 participants. The sample was divided into four age groups: 12-18, 19-25, 26-55, and 56-85. Participants were selected through convenience sampling. General coping strategies employed by these participants were assessed using a brief COPE scale, the COVID-19 fear scale was used to assess fear, and coronavirus anxiety scale was used to screen anxiety. RESULTS: Our results indicated that age has a negative correlation with adaptive coping and positive correlation with problem-focused coping, emotion-focused coping, and fear of COVID-19. The most used coping strategy among the age groups 12-18 and 19-25 age group was adaptive coping; for the 26-55 age group, it was problem-focused coping; and for the age group 56-85, emotion-focused coping was used the most. CONCLUSION: The findings of the current research could help to make the intervention procedure tailor-made for different age groups, thereby ensuring adherence and relatability to the precautionary directions.

Current Standing and Technical Guidance on Intracellular Drug Quantification: A New Site Specific Bioavailability Prediction Approach.

Determination of intracellular concentration becomes essential for the drugs having target receptors or bioactivation site inside the cells. Majority of the antiviral drugs are nucleoside analogs and their intracellular phosphorylated metabolites are active. The anticancer drugs of the cellular enzyme and nucleoside analog category are also required to be undergone intracellular drug level analysis. In this review, we have sequentially described the cell isolation protocols, cell lysis techniques and sample preparation approach to be followed for quantification of intracellular levels of selected antiviral and anticancer drugs. Major limitations for intracellular analyte quantification and their possible way out has been discussed. Currently, no literature is available summarizing these important aspects including bioanalysis of intracellular quantification of either antiviral or anticancer drugs. This review, thus, can be considered to be first of its kind and will be highly useful in providing guidance for intracellular drug analysis aiming determination of the site-specific bioavailability.

Mechanisms of Parenteral Nutrition-Associated Liver and Gut Injury.

Parenteral nutrition (PN) has revolutionized the care of patients with intestinal failure by providing nutrition intravenously. Worldwide, PN remains a standard tool of nutrition delivery in neonatal, pediatric, and adult patients. Though the benefits are evident, patients receiving PN can suffer serious cholestasis due to lack of enteral feeding and sometimes have fatal complications from liver injury and gut atrophy, including PN-associated liver disease or intestinal failure-associated liver disease. Recent studies into gut-systemic cross talk via the bile acid-regulated farnesoid X receptor (FXR)-fibroblast growth factor 19 (FGF19) axis, gut microbial control of the TGR5-glucagon-like peptide (GLP) axis, sepsis, and role of prematurity of hepatobiliary receptors are greatly broadening our understanding of PN-associated injury. It has also been shown that the composition of ω-6/ω-3 polyunsaturated fatty acids given parenterally as lipid emulsions can variably drive damage to hepatocytes and cell integrity. This manuscript reviews the mechanisms for the multifactorial pathogenesis of liver disease and gut injury with PN and discusses novel ameliorative strategies.

An Exploration of Advancement in Analytical Methodology for Quantification of Anticancer Drugs in Biomatrices.

Significant numbers of newer anticancer drugs are regularly entering into the market worldwide to fight against different types of cancers. Analytical methodologies are being developed to quantitate those molecules in a variety of matrices during their drug development stages. Selection of biological matrices for developing bioanalytical methods is based on the mechanism of action, site of action, site of metabolism and route of excretion of the drugs or their metabolites. In this review, we have described the current scenario and advancements in bioanalytical techniques for quantification of different anticancer drugs in a variety of biomatrices with a special emphasis on sample preparation techniques. We have discussed and summarized different bioanalytical aspects for anticancer drugs, which can give direction to the researcher for choosing appropriate techniques for their quantification needs.

An economic analysis of salmonella detection in fresh produce, poultry, and eggs using whole genome sequencing technology in Canada.

The study estimates the annual costs of nontyphoidal Salmonellosis (referred to as Salmonellosis from hereon) from fresh produce, poultry and eggs in Canada. It also estimates the economic benefits from introduction of Whole Genome Sequencing (WGS) in detection of Salmonellosis clusters and outbreaks. Monetary and non-monetary costs from Salmonellosis are estimated. Monetary costs are divided into direct healthcare, indirect, federal and producer costs. Probability models are used to account for uncertainty in the cost-of-illness estimates. Two types of non-monetary costs have been estimated: Disability-adjusted Live Years and Quality-adjusted Life Years. These estimates are then used to calculate the economic impact of WGS on detection of Salmonellosis. The estimated incidence of illnesses is 47,082 annually, which represents a cost of $287.78 million (total cases) and $166.28 million (reported cases) from the traditional technology. The total net benefit from introduction of WGS is estimated to range from $5.21 million-$90.25 million. All monetary values are in CAD unless stated otherwise. WGS will help in reducing the economic burden from Salmonellosis. These estimates help will aid policy related decision making.

Antimicrobial susceptibility pattern of extended-spectrum beta- lactamase producing Klebsiella pneumoniae clinical isolates in an Indian tertiary hospital.

OBJECTIVE: There is an increased prevalence of extended-spectrum beta-lactamase producing Klebsiella pneumoniae (ESBL-KP) worldwide including India, which is a major concern for the clinicians, especially in intensive care units and pediatric patients. This study aims to determine the prevalence of ESBL-KP and antimicrobial sensitivity profile to plan a proper hospital infection control program to prevent the spread of resistant strains. METHODS: KP isolates obtained from various clinical samples were evaluated to detect the production of ESBL by phenotypic methods. Antimicrobial susceptibility profile was also determined of all the isolates. FINDINGS: Of 223 nonduplicate isolates of K. pneumoniae, 114 (51.1%) were ESBL producer and antimicrobial susceptibility profile showed the isolates were uniformly sensitive to imipenem and highly susceptible to beta-lactamase inhibitor combination drugs (67-81%) and aminoglycosides (62-76%), but less susceptible to third generation cephalosporins (14-24%) and non-ß-lactam antibiotics such as nitrofurantoin (57%), fluoroquinolones (29-57%), piperacillin (19-23%), and aztreonam (15-24%). CONCLUSION: This study found that beta-lactamase inhibitor combinations are effective in treatment of such infections due to ESBL-KP thus these drugs should be a part of the empirical therapy and carbapenems should be used when the antimicrobial susceptibility tests report resistance against inhibitors combinations.