Iranian Journal of Pharmaceutical Research最新文献

{"title":"Investigating the Relationship Between the Market Orientation Approach of Pharmaceutical Companies and Their Innovative Performance: The Mediating Role of Dynamic Capabilities and Corporate Social Responsibility","authors":"J. Shirmohammadi, S. Azizi, H. Rasekh, F. Peiravian, Mahyar Polroudi Moghaddam","doi":"10.5812/ijpr-135094","DOIUrl":"https://doi.org/10.5812/ijpr-135094","url":null,"abstract":"Background: Given the intensifying competition, adapting to the market environment and meeting customer demands are crucial aspects of the evolving marketing process. Market orientation (MO) represents an organizational culture encompassing shared beliefs and values that prioritize the customer's role in business planning. Objectives: This study seeks to explore the impact of MO on innovative performance (IP) and the potential mediating role of dynamic capabilities (DC) and corporate social responsibility (CSR) in this relationship. Methods: For this study, a structured quantitative questionnaire was distributed to 100 local pharmaceutical companies, resulting in 300 completed questionnaires. Each questionnaire consisted of four main components, which were filled out by three managers from each company: Chief executive officer (CEO), marketing manager, and research and development manager. The collected data were analyzed using SPSS software and structural equation methods to examine the research questions and hypotheses. Results: According to the study findings, there was a positive correlation between employee age, organizational structure, sales volume, and the presence of private companies with IP. MO, DC, and CSR showed a direct and significant relationship with IP. Moreover, the CSR of the company influenced IP through the mediating role of DC. Market orientation was found to enhance explorative IP, leading to improvements in existing processes and services. Conclusions: Based on the study results, it was found that MO has a direct positive impact on IP, leading to improvements in the company's existing processes through its influence on exploratory performance.","PeriodicalId":14595,"journal":{"name":"Iranian Journal of Pharmaceutical Research","volume":"80 1","pages":""},"PeriodicalIF":1.6,"publicationDate":"2023-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79258614","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Interaction of SRL-2 Peptide with LRP-1 Receptor and Identification of Breast Cancer Related Biomarkers: An In-silico Approach","authors":"Yasamin Davatgaran-Taghipour, Yousof Saeedi-Honar, Roya Salehi, Amir Zarebkohan, Vladimir P.Torchilin","doi":"10.5812/ijpr-136624","DOIUrl":"https://doi.org/10.5812/ijpr-136624","url":null,"abstract":"Background: Breast cancer is a multifaceted disease characterized by genetic and epigenetic changes that lead to uncontrolled cell growth and metastasis. Early detection and treatment are crucial for managing diseases. Objectives: The objective of this study is to investigate the potential of chimeric peptides for drug delivery and to identify biomarkers associated with breast cancer. Recent studies have shown that the low-density lipoprotein receptor-related protein 1 (LRP-1) receptor has a significant impact on the development of breast cancer. In order to facilitate the identification of biomarkers, we have created a chimeric peptide that has been proven to bind successfully to the LRP-1 receptor. Methods: To identify biomarkers, we utilized advanced computational methods to conduct a meta-analysis of microarray data. Specifically, the g:Profiler and eXpression2Kinases (X2K) databases were utilized to identify gene ontologies and transcription factors. We then used the Human Protein Atlas to identify and assess crucial gene expressions. Results: Our results demonstrated that nucleolar and spindle-associated protein 1 (NUSAP1), melatonin receptor 1A (MELT), and cyclin-dependent kinase 1 (CDK1) are three hub genes that play pivotal roles in the pathogenesis of breast cancer. Conclusions: The research findings provide a deeper understanding of the molecular mechanisms involved in developing breast cancer. These findings have significant implications for developing novel therapies and diagnostics for this disease.","PeriodicalId":14595,"journal":{"name":"Iranian Journal of Pharmaceutical Research","volume":"40 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135004717","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Interaction of SRL-2 Peptide with LRP-1 Receptor and Identification of Breast Cancer Related Biomarkers: An In-silico Approach","authors":"Yasamin Davatgaran-Taghipour, Yousef Saeedi-Honar, R. Salehi, A. Zarebkohan, Vladimir P.Torchilin","doi":"10.5812/ijpharm-136624","DOIUrl":"https://doi.org/10.5812/ijpharm-136624","url":null,"abstract":"Background: Breast cancer is a multifaceted disease characterized by genetic and epigenetic changes that lead to uncontrolled cell growth and metastasis. Early detection and treatment are crucial for managing diseases. Objectives: The objective of this study is to investigate the potential of chimeric peptides for drug delivery and to identify biomarkers associated with breast cancer. Recent studies have shown that the low-density lipoprotein receptor-related protein 1 (LRP-1) receptor has a significant impact on the development of breast cancer. In order to facilitate the identification of biomarkers, we have created a chimeric peptide that has been proven to bind successfully to the LRP-1 receptor. Methods: To identify biomarkers, we utilized advanced computational methods to conduct a meta-analysis of microarray data. Specifically, the g:Profiler and eXpression2Kinases (X2K) databases were utilized to identify gene ontologies and transcription factors. We then used the Human Protein Atlas to identify and assess crucial gene expressions. Results: Our results demonstrated that nucleolar and spindle-associated protein 1 (NUSAP1), melatonin receptor 1A (MELT), and cyclin-dependent kinase 1 (CDK1) are three hub genes that play pivotal roles in the pathogenesis of breast cancer. Conclusions: The research findings provide a deeper understanding of the molecular mechanisms involved in developing breast cancer. These findings have significant implications for developing novel therapies and diagnostics for this disease.","PeriodicalId":14595,"journal":{"name":"Iranian Journal of Pharmaceutical Research","volume":"46 1","pages":""},"PeriodicalIF":1.6,"publicationDate":"2023-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82580362","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis, Antioxidant, Cytotoxicity, Induce Apoptosis Investigation and Docking Study of New Halogenated Dihydropyrano[3,2-b]Chromene-3-Carbonitrile Derivatives on MCF-7 Breast Cancer Cell Line","authors":"Touba Eslaminejad, Ehsan Faghih Mirzaei, Mehdi Abaszadeh","doi":"10.5812/ijpr-132932","DOIUrl":"https://doi.org/10.5812/ijpr-132932","url":null,"abstract":"Background: Chromene derivatives showed numerous biological activities. In the current study, the antioxidant, cytotoxicity, and apoptosis properties of halogenated dihydropyrano[3,2-b]chromene-3-carbonitrile derivatives (HDCCD) on MCF-7 cell line have been examined. Objectives: This study's principal point was synthesizing new halogenated pyranochromene derivatives and assessing their cytotoxic effects and apoptosis potential on MCF-7 breast cancer cell line by flow cytometry. Methods: Initially, 6-chloro- and 6-bromo-3-hydroxychromone compounds were prepared. In the next step, a series of HDCCD were synthesized by a one-pot three-component reaction of these two compounds, aromatic aldehydes, and malononitrile, in the presence of triethylamine in EtOH at reflux conditions. These compounds were fully characterized by standard spectroscopic techniques (IR, 1H, and 13C NMR) and elemental analyses. The potential of the antioxidant activity was determined by using ferric reducing antioxidant power assay (FRAP). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) were used to evaluate metabolic activity. The nitric oxide (NO) and malondialdehyde (MDA) biomarkers of the exposed cells were evaluated on the cells and their supernatant. To quantify apoptotic death of MCF-7 breast cancer cells treated by the compounds at their IC50 concentrations, Annexin V-FITC apoptosis detection kit was utilized. Molecular docking of compounds (6a-j) into the Cyclin-dependent kinase 6 (PDB code: 4EZ5) was carried out, and the probable binding mode of compounds 6e and 6j was determined. Results: A dose-response relationship was seen in all the compounds. Most of them induced cytotoxic effects on the cells. Nitrite concentration of the culture media of the cells was decreased compared to the control. Malondialdehyde levels of the cells were below the range of the control by the addition of 6b, 6d, 6e, 6f, and 6g compounds on the cells, while the addition of the 6a, 6c, 6h, 6i, and 6j compounds increased the MDA level compared to the control. Flow cytometric analysis showed that most of the exposed cells were in the early and late apoptotic stage, and a few of them were in the necrotic stage. Conclusions: It could be concluded that HDCCD (6a-j) was toxic and caused death in the cells by apoptosis. The compounds have lipophilic characteristics, so they can easily pass the cell membrane. As confirmed by LDH results, it can be concluded that the cytotoxicity is connected with apoptosis rather than necrosis, endorsed by flowcytometry analysis afterward.","PeriodicalId":14595,"journal":{"name":"Iranian Journal of Pharmaceutical Research","volume":"76 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135972030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hepatotoxicity Induced by Azole Antifungal Agents: A Review Study.","authors":"Amin Rakhshan, Bardia Rahmati Kamel, Ali Saffaei, Maria Tavakoli-Ardakani","doi":"10.5812/ijpr-130336","DOIUrl":"10.5812/ijpr-130336","url":null,"abstract":"<p><strong>Context: </strong>Fungal infections are very common, and several medications are used to treat them. Azoles are prescribed widely to treat fungal infections. In addition to therapeutic effects, any drug can be accompanied by side effects in patients. One of the most important complications in this regard is liver injury. Therefore, hepatotoxicity induced by azole antifungal drugs were reviewed in this study.</p><p><strong>Evidence acquisition: </strong>English scientific papers were evaluated to review the effects of hepatotoxicity by azole antifungal agents, and the related studies' results were summarized using a table. The systematic search was implemented on electronic databases, including PubMed, Google Scholar, and Science Direct. Original articles and review articles that were published before April 1, 2022, were included in the study. Those articles without available full text or non-English articles were excluded. Also, articles that reported pediatric data were excluded.</p><p><strong>Results: </strong>Most studies have reported the effects of hepatotoxicity by azole antifungal agents, and their mechanisms have been described.</p><p><strong>Conclusions: </strong>Clinical evaluations regarding the hepatotoxicity of antifungal agents provided in the literature were reviewed. Therefore, it is recommended to prescribe these drugs with caution in high-risk patients suffering from liver diseases, and patients should be monitored for hepatotoxicity. However, more research is needed to evaluate the hepatotoxicity of azole antifungal agents and select appropriate drugs according to cost-effectiveness and the side effects' profiles, relying on lower incidence of this liver complication.</p>","PeriodicalId":14595,"journal":{"name":"Iranian Journal of Pharmaceutical Research","volume":"1 1","pages":"e130336"},"PeriodicalIF":1.6,"publicationDate":"2023-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10728840/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87890036","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigating the Changes in Cream Properties Following Topical Application and Their Influence on the Product Efficiency.","authors":"Nadia Salehi,&nbsp;Seyedeh Maryam Mortazavi,&nbsp;Hamidreza Moghimi","doi":"10.5812/ijpr.123946","DOIUrl":"https://doi.org/10.5812/ijpr.123946","url":null,"abstract":"<p><p>Topical products are not stable following application to the skin due to the evaporation of volatile components. Such changes have been demonstrated in liquid emulsions, but there is almost no study available for creams in this respect. The aim of the present investigation is to evaluate the changes in cream properties following topical application and their influence on product efficiency. A method has also been designed and validated to mimic cream application to the skin. To perform this investigation, five different creams were prepared and alterations of type of creams, size of droplets of the dispersed phase, occlusivity, water content and rate of water loss were studied after application. These changes were then attributed to the type of cream, water content, presence of humectant, and time post application. The results demonstrated that creams changed intensely after application, including the phase inversion of O/W formulations, changes in the occlusivity of creams, reduction of water content, rate of water evaporation and droplet size. Such changes could be controlled partly by humectants. The present results suggest that formulators should be aware of such possible changes and required precautions should be taken in advance.</p>","PeriodicalId":14595,"journal":{"name":"Iranian Journal of Pharmaceutical Research","volume":"21 1","pages":"e123946"},"PeriodicalIF":1.6,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/60/6a/ijpr-21-1-123946.PMC10024334.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9155561","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Impact of Process and Formulation Parameters on the Fabrication of Efavirenz Nanosuspension to Improve Drug Solubility and Dissolution.","authors":"Mahtab Rashed,&nbsp;Simin Dadashzadeh,&nbsp;Noushin Bolourchian","doi":"10.5812/ijpr-129409","DOIUrl":"https://doi.org/10.5812/ijpr-129409","url":null,"abstract":"<p><strong>Background: </strong>Efavirenz nanosuspensions (EZ-NSs) were developed by the wet milling method as the most promising top-down nanosizing technique. Different process and formulation parameters were studied and optimized to produce appropriate EZ-NS in suitable conditions to enhance drug dissolution.</p><p><strong>Methods: </strong>In the preliminary studies, various polymeric stabilizers, including Pluronic F68, sodium carboxymethylcellulose (CMC), hydroxypropyl methylcellulose (HPMC), and polyvinyl alcohol (PVA), as well as different sizes and weight of milling beads were used to prepare NSs. The effect of sodium lauryl sulfate (SLS) concentration on the NS properties was also evaluated. The influence of other formulation and process parameters, including polymer concentration, milling speed, and milling time, on the particle size and distribution of NSs were investigated using Box-Behnken design. The optimized freeze-dried nanosuspension was characterized by redispersibility, physicochemical properties, and stability.</p><p><strong>Results: </strong>A combination of PVA and SLS was selected as steric and electrostatic stabilizers. The optimum EZ-NS displayed a uniform size distribution with a mean particle size and zeta potential of 254.4 nm and 21.1 mV, respectively. The solidified nanosuspension was well redispersed to the original nanoparticles. Significantly enhanced aqueous solubility (about 11-fold) and accelerated dissolution rate were observed for the optimized formulation. This could be attributed to the reduced particle size and partial amorphization of EZ during the preparation process, studied by X-ray diffraction. Accelerated studies confirmed the stability of the optimum freeze-dried formulation over the examined period of three months.</p><p><strong>Conclusions: </strong>Optimization of different variables led to the formation of EZ-NSs with desired properties through wet milling in a very short time compared to the previous study and therefore reduced production costs. This formulation seems to be a suitable approach for solubility and dissolution enhancement of EZ and may have a great potential to improve the drug's oral bioavailability.</p>","PeriodicalId":14595,"journal":{"name":"Iranian Journal of Pharmaceutical Research","volume":"21 1","pages":"e129409"},"PeriodicalIF":1.6,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/de/56/ijpr-21-1-129409.PMC10024318.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9155563","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improvement of Phenolic Compound Extraction by Using Ion Exchange Chromatography and Evaluation of Biological Activities of Polyphenol-enriched Fraction of <i>Rosa canina</i> Fruits.","authors":"Zahra Sabahi,&nbsp;Seyed Muhammad Farid Hasan,&nbsp;Seyed Abdulmajid Ayatollahi,&nbsp;Fatemeh Farmani,&nbsp;Afshin Afsari,&nbsp;Mahmoodreza Moein","doi":"10.5812/ijpr-126558","DOIUrl":"https://doi.org/10.5812/ijpr-126558","url":null,"abstract":"<p><strong>Background: </strong><i>Rosa canina</i> has been traditionally known as a medicinal plant. Different applications of fruits (Rose hip) comprise the food, perfume, and cosmetic industries.</p><p><strong>Objectives: </strong>This study aimed to prepare an enriched polyphenolic fraction from <i>Rosa canina</i> in addition to its biological activities.</p><p><strong>Methods: </strong>Poly phenolic enriched fraction was prepared using Amberlite XAD-7 for removing unwanted components. Phenols, flavonoids, and anthocyanins content analyses showed that they increased significantly compared to the extract. HPLC analysis showed that this fraction is a rich source of ascorbic acid.</p><p><strong>Results: </strong>The results of the DPPH, ferric-reducing antioxidant power (FRAP), ABTS, and nitric oxide assay confirmed that the antioxidant activities of the fraction had been increased compared to the extract. The oxygen radical absorbance capacity (ORAC) assay and cellular antioxidant activity of the fraction also confirmed its potential antioxidant activity. This fraction showed xanthine oxidase inhibitory activity at 100 µg/mL concentration. Comet assay analysis revealed that this fraction at 25 - 100 µg/mL concentrations inhibited H₂O₂ genotoxicity in human lymphocytes.</p><p><strong>Conclutions: </strong>This study suggests that the fruit of <i>Rosa canina</i> could be considered as a potential antioxidant, a xanthine oxidase inhibitor, and an antigenotoxic source, and the application of Amberlite XAD-7 improves extraction efficiencies through enrichment of phenolic compounds in this plant.</p>","PeriodicalId":14595,"journal":{"name":"Iranian Journal of Pharmaceutical Research","volume":"21 1","pages":"e126558"},"PeriodicalIF":1.6,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/a9/07/ijpr-21-1-126558.PMC10024319.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9155564","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation of the Anticoagulant and Procoagulant Fractions of the Venom of Iranian Endemic <i>Echis carinatus</i>.","authors":"Nafiseh Nasri Nasrabadi,&nbsp;Nasser Mohammadpour Dounighi,&nbsp;Minoo Ahmadinejad,&nbsp;Hadi Rabiei,&nbsp;Maryam Tabarzad,&nbsp;Mojtaba Najafi,&nbsp;Hossein Vatanpour","doi":"10.5812/ijpr-127240","DOIUrl":"https://doi.org/10.5812/ijpr-127240","url":null,"abstract":"<p><strong>Background: </strong>The venom of <i>Echis carinatus</i> contains both procoagulant and anticoagulant components that can either promote or block the blood coagulation cascade, and some of these components affect platelet function in different ways.</p><p><strong>Objectives: </strong>The present study focuses on setting up a procedure for the purification of crude venom and designing appropriate clotting tests in order to characterize the procoagulant and anticoagulant fractions of <i>E. carinatus</i> venom.</p><p><strong>Methods: </strong>Chromatographic methods, including gel filtration, ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC), were applied for purifying these fractions. Coagulant activity testing, prothrombin time (PT), and activated partial thromboplastin time (APTT) were used to determine procoagulant and anticoagulant properties. For measuring molecular weight, 15% SDS-PAGE electrophoresis with a molecular weight standard ranging from 6.5 to 200 kDa was used.</p><p><strong>Results: </strong>We obtained five fractions named F₁, F₂, F₃, F₄, and F₅. The F₁ and F₂ fractions showed procoagulant activity, and the F₅ fraction had anticoagulant activity. The molecular weight of F_2.4.2 from fraction F₂ and F_5.1 from fraction F₅ were analyzed by SDS-PAGE electrophoresis under the reducing condition. These factors were identified as a single protein band at the end of purification. The molecular weights of these purified fractions were estimated to be 7.5 kDa and 38 kDa for F_5.1(b) and F_2.4.2(b), respectively.</p><p><strong>Conclusions: </strong>Our findings suggest an efficient and suitable procedure for the identification and purification of the procoagulant and anticoagulant factors of the venom of Iranian <i>E. carinatus</i> using the PT and APTT assays.</p>","PeriodicalId":14595,"journal":{"name":"Iranian Journal of Pharmaceutical Research","volume":"21 1","pages":"e127240"},"PeriodicalIF":1.6,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/e8/fa/ijpr-21-1-127240.PMC10024320.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9163589","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Targeting Caspase-3 Gene in rCHO Cell Line by CRISPR/Cas9 Editing Tool and Its Effect on Protein Production in Manipulated Cell Line.","authors":"Amirabbas Rahimi,&nbsp;Morteza Karimipoor,&nbsp;Reza Mahdian,&nbsp;Atefeh Alipour,&nbsp;Sadi Hosseini,&nbsp;Hooman Kaghazian,&nbsp;Abdolrahim Abbasi,&nbsp;Hosein Shahsavarani,&nbsp;Mohammad Ali Shokrgozar","doi":"10.5812/ijpr-130236","DOIUrl":"https://doi.org/10.5812/ijpr-130236","url":null,"abstract":"<p><strong>Background: </strong>Chinese hamster ovary (CHO) cells are the widely used mammalian cell host for biopharmaceutical manufacturing. During cell cultures, CHO cells lose viability mainly from apoptosis. Inhibiting cell death is useful because prolonging cell lifespans can direct to more productive cell culture systems for biotechnology requests.</p><p><strong>Objectives: </strong>This study exploited a CRISPR/Cas9 technology to generate site-specific gene disruptions in the caspase-3 gene in the apoptosis pathway, which acts as an apoptotic regulator to extend cell viability in the CHO cell line.</p><p><strong>Methods: </strong>The STRING database was used to identify the key pro-apoptotic genes to be modified by CRISPR/Cas9 system. The guide RNAs targeting the caspase-3 gene were designed, and vectors containing sgRNA and Cas9 were transfected into CHO cells that expressed erythropoietin as a heterologous protein. Indel formation was investigated by DNA sequencing. Caspase-3 expression was quantified by real-time PCR and western blot. The effect of editing the caspase-3 gene on the inhibition of apoptosis was also investigated by induction of apoptosis in manipulated cell lines by oleuropein. Finally, the erythropoietin production in the edited cells was compared to the control cells.</p><p><strong>Results: </strong>The caspase-3 manipulation significantly prolongation of the cell viability and decreased the caspase-3 expression level of protein in manipulated CHO cells (more than 6-fold, P-value < 0.0001). Manipulated cells displayed higher threshold tolerance to apoptosis compared to the control cells when they were induced by oleuropein. They show a higher IC50 than the control ones (7271 µM/mL Vs. 5741 µM/mL). They also show a higher proliferation rate than the control cells in the presence of an apoptosis inducer (P-value < 0.0001). Furthermore, manipulated cell lines significantly produce more recombinant protein in the presence of 2,000 µM oleuropein compared to the control ones (P-value = 0.0021).</p><p><strong>Conclusions: </strong>We understood that CRISPR/Cas9 could be effectively applied to suppress the expression of the caspase-3 gene and rescue CHO cells from apoptosis induced by cell stress and metabolites. The CRISPR/Cas9 system-assisted caspase-3 gene ablation can potentially increase erythropoietin yield in CHO cells.</p>","PeriodicalId":14595,"journal":{"name":"Iranian Journal of Pharmaceutical Research","volume":"21 1","pages":"e130236"},"PeriodicalIF":1.6,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/c9/19/ijpr-21-1-130236.PMC10007989.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9106077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}