Pub Date : 2025-02-01DOI: 10.1016/j.xphs.2024.11.008
Clara Hartmanshenn , Alexander Bechtold , Thomas Kwok , Jeff Mora , Nathan Contrella , Alex Confer , Rachel Bade , Teresa Andreani , Jonathan M.E. Hughes , Billy Chen , Eric Sirota , Lorenzo Codan , David J. Lamberto , Yingju Xu , Nastaran Salehi , Stephen Crowley
To make investigational drug candidates available to patients sooner, timelines for drug development are becoming shorter. Synthesis route scouting for active pharmaceutical ingredients (API) and drug product development often must occur simultaneously, requiring formulators to make decisions regarding drug product process selection before commercial API route finalization. Alternatively, the formulation strategy may be locked, thereby constraining drug substance processes with strict API attribute requirements. Critical quality attributes of the drug product can depend heavily on the API, yet final physical attributes may not be known early on in development. Furthermore, the desire to reduce pill burden means higher drug loading in formulations, leaving little room for excipients to compensate for suboptimal API performance. The opposing challenges of API synthetic route and drug product formulation development typically lead to elongated development timelines requiring an iterative approach. In this work, a coordinated strategy was designed and implemented to deliberately range API attributes via crystallization and milling techniques to enable robust assessment of downstream manufacturing and significantly reduce the time for final process selection. The study presented was conducted on a protease inhibitor targeted for treatment of Covid-19. Given the emergent need for treatment options, this dramatically accelerated approach was crucial for potential emergency use authorization (EUA).
{"title":"Attribute ranging as a coordinated strategy between drug substance and drug product to accelerate commercial process nomination","authors":"Clara Hartmanshenn , Alexander Bechtold , Thomas Kwok , Jeff Mora , Nathan Contrella , Alex Confer , Rachel Bade , Teresa Andreani , Jonathan M.E. Hughes , Billy Chen , Eric Sirota , Lorenzo Codan , David J. Lamberto , Yingju Xu , Nastaran Salehi , Stephen Crowley","doi":"10.1016/j.xphs.2024.11.008","DOIUrl":"10.1016/j.xphs.2024.11.008","url":null,"abstract":"<div><div>To make investigational drug candidates available to patients sooner, timelines for drug development are becoming shorter. Synthesis route scouting for active pharmaceutical ingredients (API) and drug product development often must occur simultaneously, requiring formulators to make decisions regarding drug product process selection before commercial API route finalization. Alternatively, the formulation strategy may be locked, thereby constraining drug substance processes with strict API attribute requirements. Critical quality attributes of the drug product can depend heavily on the API, yet final physical attributes may not be known early on in development. Furthermore, the desire to reduce pill burden means higher drug loading in formulations, leaving little room for excipients to compensate for suboptimal API performance. The opposing challenges of API synthetic route and drug product formulation development typically lead to elongated development timelines requiring an iterative approach. In this work, a coordinated strategy was designed and implemented to deliberately range API attributes via crystallization and milling techniques to enable robust assessment of downstream manufacturing and significantly reduce the time for final process selection. The study presented was conducted on a protease inhibitor targeted for treatment of Covid-19. Given the emergent need for treatment options, this dramatically accelerated approach was crucial for potential emergency use authorization (EUA).</div></div>","PeriodicalId":16741,"journal":{"name":"Journal of pharmaceutical sciences","volume":"114 2","pages":"Pages 1010-1016"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142791838","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.xphs.2024.10.012
Robbe Van Pottelberge , Roman Matthessen , Shauna Salem , Ben Goffin , Nancee Oien , Pratima Bharti , David Ripley
Ribonucleases (RNases) are ubiquitous in nature, being able to cleave a wide range of polyribonucleotides. While the presence of microbial and viral contamination in sterile manufacturing is highly studied and controlled, there are no standardized practices for evaluating RNase in the production facility. Since the COVID-19 pandemic, mRNA-LNP based vaccines have become part of routine large-scale manufacturing. The unstable nature of mRNA poses new challenges to safeguard the working efficacy of mRNA – Lipid nanoparticle (LNP) based vaccines or therapeutics, where the presence of RNase in the formulation process could have a profound impact on the mRNA integrity. In this article, lessons learned are presented with respect to the evaluation of RNase contamination during LNP drug product formulation and analysis. Using sensitive detection methods, the potential presence of RNase in the manufacturing of mRNA-LNPs was investigated. Additionally, capillary gel electrophoresis (CGE) data, used to measure mRNA integrity, demonstrate the quality of the active mRNA substance and importance of suitable RNase control strategies. The results and cases presented in this paper should pave the way forward for evaluation and control strategies dedicated to mRNA-LNP based vaccines and therapeutics.
{"title":"Importance of RNase monitoring during large-scale manufacturing and analysis of mRNA-LNP based vaccines","authors":"Robbe Van Pottelberge , Roman Matthessen , Shauna Salem , Ben Goffin , Nancee Oien , Pratima Bharti , David Ripley","doi":"10.1016/j.xphs.2024.10.012","DOIUrl":"10.1016/j.xphs.2024.10.012","url":null,"abstract":"<div><div>Ribonucleases (RNases) are ubiquitous in nature, being able to cleave a wide range of polyribonucleotides. While the presence of microbial and viral contamination in sterile manufacturing is highly studied and controlled, there are no standardized practices for evaluating RNase in the production facility. Since the COVID-19 pandemic, mRNA-LNP based vaccines have become part of routine large-scale manufacturing. The unstable nature of mRNA poses new challenges to safeguard the working efficacy of mRNA – Lipid nanoparticle (LNP) based vaccines or therapeutics, where the presence of RNase in the formulation process could have a profound impact on the mRNA integrity. In this article, lessons learned are presented with respect to the evaluation of RNase contamination during LNP drug product formulation and analysis. Using sensitive detection methods, the potential presence of RNase in the manufacturing of mRNA-LNPs was investigated. Additionally, capillary gel electrophoresis (CGE) data, used to measure mRNA integrity, demonstrate the quality of the active mRNA substance and importance of suitable RNase control strategies. The results and cases presented in this paper should pave the way forward for evaluation and control strategies dedicated to mRNA-LNP based vaccines and therapeutics.</div></div>","PeriodicalId":16741,"journal":{"name":"Journal of pharmaceutical sciences","volume":"114 2","pages":"Pages 1520-1528"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142468293","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Acyl glucuronide (AG) is a reactive metabolite that causes idiosyncratic drug toxicity (IDT). Although the instability of AG is used to predict the IDT risk of novel drug candidates, it sometimes overestimates the IDT risk. We investigated whether the rate of enzymatic AG hydrolysis in human liver microsomes (HLM) can predict the risk of IDT. We used 16 drugs classified into three categories in terms of IDT risk: drugs withdrawn from the market owing to severe IDT (withdrawn, WDN) and drugs still being on the market, regardless of IDT risk (warning, WA) or not (safe, SA). AG was incubated with HLM, and the resulting parent drugs for AG hydrolysis were quantified using HPLC. The rate of enzymatic AG hydrolysis in the HLM of WDN was higher than that in WA and SA, and no difference was observed between WA and SA. We categorized WA and SA as commercially available (CA) drugs and performed a logistic regression analysis. The rate of enzymatic AG hydrolysis in HLM significantly distinguished WDN drugs from CA drugs, with an estimated classification value of 0.189 nmol/min/mg protein. In conclusion, the rate of enzymatic AG hydrolysis in HLM may be useful for predicting the risk in drug development.
{"title":"Enzymatic hydrolysis of acyl glucuronide metabolites in human liver microsomes correlates to the risk of idiosyncratic drug toxicity","authors":"Hiroaki Shimada, Hiroyuki Ikuta, Yu Hashimoto, Yusuke Yabuuchi, Atsushi Kawase, Sumio Matzno, Masahiro Iwaki","doi":"10.1016/j.xphs.2025.01.014","DOIUrl":"10.1016/j.xphs.2025.01.014","url":null,"abstract":"<div><div>Acyl glucuronide (AG) is a reactive metabolite that causes idiosyncratic drug toxicity (IDT). Although the instability of AG is used to predict the IDT risk of novel drug candidates, it sometimes overestimates the IDT risk. We investigated whether the rate of enzymatic AG hydrolysis in human liver microsomes (HLM) can predict the risk of IDT. We used 16 drugs classified into three categories in terms of IDT risk: drugs withdrawn from the market owing to severe IDT (withdrawn, WDN) and drugs still being on the market, regardless of IDT risk (warning, WA) or not (safe, SA). AG was incubated with HLM, and the resulting parent drugs for AG hydrolysis were quantified using HPLC. The rate of enzymatic AG hydrolysis in the HLM of WDN was higher than that in WA and SA, and no difference was observed between WA and SA. We categorized WA and SA as commercially available (CA) drugs and performed a logistic regression analysis. The rate of enzymatic AG hydrolysis in HLM significantly distinguished WDN drugs from CA drugs, with an estimated classification value of 0.189 nmol/min/mg protein. In conclusion, the rate of enzymatic AG hydrolysis in HLM may be useful for predicting the risk in drug development.</div></div>","PeriodicalId":16741,"journal":{"name":"Journal of pharmaceutical sciences","volume":"114 2","pages":"Pages 1307-1314"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143052930","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The antigen-targeting to dendritic cells (DCs) has gained increasing attention as the potential approach for immunotherapy in recent years due to the ability of DCs to regulate innate and adaptive immunity. In the present study, the immunogenicity and protective efficiency of mannan-decorated PLGA nanoparticles (NPs) loaded with multi-epitopes mycobacterium tuberculosis antigen (HspX-Ppe44-EsxV) were evaluated as a targeted delivery system to DCs. For this purpose, PLGA nanoparticle formulations were prepared and subsequently decorated by mannan. The physicochemical properties and level of mannan incorporation, as well as encapsulation efficiency and antigen release, were assessed. The potential of formulated NPs for antigen targeting to DCs, and immunogenicity against tuberculosis (TB) were investigated using immunofluorescence assay and in-vivo experiments. Mannan incorporation enhanced the uptake of fusion-loaded PLGA by DCs. The cytokine and antibody assays demonstrated that mannosylation of NPs and BCG-primed mice boosted by mannan-PLGA could significantly elevate Th1-biased immune responses relative to the BCG and non-modified PLGA NPs. Our findings also proved that the mannosylated vaccine in the presence of CpG could evoke Th1 and Th17 responses with appropriate protective efficiency against TB in mice. This result illustrated that the active targeting of DCs by mannan-PLGA NPs could induce a proper anti-tuberculosis response, which is essential for protection against tuberculosis.
{"title":"Assessment of immunogenicity and protective efficiency of multi-epitope antigen-loaded in mannan decorated PLGA nanoparticles against tuberculosis","authors":"Yousef Amini , Mona Kabiri , Saeid Amel Jamehdar , Mojtaba Sankian , Zahra Meshkat , Sirwan Zare , Saman Soleimanpour , Hadi Farsiani , Bagher Moradi , Mohsen Tafaghodi","doi":"10.1016/j.xphs.2024.11.025","DOIUrl":"10.1016/j.xphs.2024.11.025","url":null,"abstract":"<div><div>The antigen-targeting to dendritic cells (DCs) has gained increasing attention as the potential approach for immunotherapy in recent years due to the ability of DCs to regulate innate and adaptive immunity. In the present study, the immunogenicity and protective efficiency of mannan-decorated PLGA nanoparticles (NPs) loaded with multi-epitopes mycobacterium tuberculosis antigen (HspX-Ppe44-EsxV) were evaluated as a targeted delivery system to DCs. For this purpose, PLGA nanoparticle formulations were prepared and subsequently decorated by mannan. The physicochemical properties and level of mannan incorporation, as well as encapsulation efficiency and antigen release, were assessed. The potential of formulated NPs for antigen targeting to DCs, and immunogenicity against tuberculosis (TB) were investigated using immunofluorescence assay and <em>in-vivo</em> experiments. Mannan incorporation enhanced the uptake of fusion-loaded PLGA by DCs. The cytokine and antibody assays demonstrated that mannosylation of NPs and BCG-primed mice boosted by mannan-PLGA could significantly elevate Th1-biased immune responses relative to the BCG and non-modified PLGA NPs. Our findings also proved that the mannosylated vaccine in the presence of CpG could evoke Th1 and Th17 responses with appropriate protective efficiency against TB in mice. This result illustrated that the active targeting of DCs by mannan-PLGA NPs could induce a proper anti-tuberculosis response, which is essential for protection against tuberculosis.</div></div>","PeriodicalId":16741,"journal":{"name":"Journal of pharmaceutical sciences","volume":"114 2","pages":"Pages 1133-1141"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142780484","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Freeze-drying is used to prolong the shelf life of pharmaceutical formulations stored in vials. To achieve this, formulations are first frozen and then dried, yielding a porous product that can in some cases be stored even at ambient conditions. In this work, the effect of different process parameters on the properties of the porous micro-structure obtained when freeze-drying dextran solutions was studied. To characterize the pore sizes, the samples were imaged with scanning electron microscopy (SEM) and the images were manually analyzed to determine the pore size distribution. To study the robustness of such manual pore characterization methodology, a reliability analysis was carried out, which showed that defining a set of guidelines leads to comparable pore size distributions among multiple participants conducting the analysis. The pore characterization methodology was then applied to products that were freeze-dried under different conditions. Higher dextran concentrations and higher cooling rates were found to lead to predominantly smaller pore sizes and longer primary drying. The conclusions of this work complement the existing literature in demonstrating the robustness of the manual pore size analysis and give valuable insight into the link between the micro-structure formed during the freezing of dextran solutions and the drying performance.
{"title":"The impact of process parameters on the lyophilized porous micro-structure: A case study of dextran","authors":"Andraž Košir , Fiora Artusio , Leif-Thore Deck , Roberto Pisano , Marco Mazzotti","doi":"10.1016/j.xphs.2024.12.020","DOIUrl":"10.1016/j.xphs.2024.12.020","url":null,"abstract":"<div><div>Freeze-drying is used to prolong the shelf life of pharmaceutical formulations stored in vials. To achieve this, formulations are first frozen and then dried, yielding a porous product that can in some cases be stored even at ambient conditions. In this work, the effect of different process parameters on the properties of the porous micro-structure obtained when freeze-drying dextran solutions was studied. To characterize the pore sizes, the samples were imaged with scanning electron microscopy (SEM) and the images were manually analyzed to determine the pore size distribution. To study the robustness of such manual pore characterization methodology, a reliability analysis was carried out, which showed that defining a set of guidelines leads to comparable pore size distributions among multiple participants conducting the analysis. The pore characterization methodology was then applied to products that were freeze-dried under different conditions. Higher dextran concentrations and higher cooling rates were found to lead to predominantly smaller pore sizes and longer primary drying. The conclusions of this work complement the existing literature in demonstrating the robustness of the manual pore size analysis and give valuable insight into the link between the micro-structure formed during the freezing of dextran solutions and the drying performance.</div></div>","PeriodicalId":16741,"journal":{"name":"Journal of pharmaceutical sciences","volume":"114 2","pages":"Pages 1434-1443"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142921668","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.xphs.2025.01.009
Syeda Yamna Zia , Sofia Ahmed , Hafiza Sumaiyya Jamal , Mehvish Perveen , Muhammad Ali Sheraz , Zubair Anwar , Syed Abid Ali
Collagenases are enzymes that break down collagen and are used in wound healing and treating various disorders. Currently, collagenase is commercially available in only ointment and injectable forms and is sensitive to various environmental factors. In the present study, different hydrogel formulations of collagenase have been prepared at pH 6.5 using carboxymethylcellulose sodium and zinc acetate with and without humectants such as propylene glycol (PG) and glycerin (GL) in varying concentrations. The formulated gels were stored at room temperature (25±2°C, 60±5% RH) and refrigerator temperature (5±3°C) for six months to evaluate their physical and up to six years for chemical stability. The gels were subjected to various tests, including organoleptic studies, spreadability, moisture content, swelling index, swelling/de-swelling, syneresis, viscosity, gelation time, and weight variation. The purity and molecular weight of collagenase have been determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS–PAGE). At the same time, its activity during the storage period was evaluated by gelatin zymography. Casein zymography was also performed to detect any caseinase contamination in the formulations. The release of the enzyme from different gel formulations was assessed using the Franz diffusion apparatus and analyzed by gelatin zymography. The results showed some physical changes that were more prominent in gels stored at room temperature than those kept refrigerated. The difference in humectant concentration was also found to affect the stability of gels. PG was found to be a better humectant than GL, particularly in a concentration of 25%. The zymography results indicated that collagenase was stable in all formulations kept in the refrigerator. In contrast, its complete degradation was noted in the preparations stored at room temperature within a month. The data generated in this study will help the formulators to commercialize a relatively economical gel formulation of collagenase that is highly stable for up to six years at refrigerator temperature (5±3°C).
{"title":"Formulation development of highly stable collagenase-containing hydrogels for wound healing","authors":"Syeda Yamna Zia , Sofia Ahmed , Hafiza Sumaiyya Jamal , Mehvish Perveen , Muhammad Ali Sheraz , Zubair Anwar , Syed Abid Ali","doi":"10.1016/j.xphs.2025.01.009","DOIUrl":"10.1016/j.xphs.2025.01.009","url":null,"abstract":"<div><div>Collagenases are enzymes that break down collagen and are used in wound healing and treating various disorders. Currently, collagenase is commercially available in only ointment and injectable forms and is sensitive to various environmental factors. In the present study, different hydrogel formulations of collagenase have been prepared at pH 6.5 using carboxymethylcellulose sodium and zinc acetate with and without humectants such as propylene glycol (PG) and glycerin (GL) in varying concentrations. The formulated gels were stored at room temperature (25±2°C, 60±5% RH) and refrigerator temperature (5±3°C) for six months to evaluate their physical and up to six years for chemical stability. The gels were subjected to various tests, including organoleptic studies, spreadability, moisture content, swelling index, swelling/de-swelling, syneresis, viscosity, gelation time, and weight variation. The purity and molecular weight of collagenase have been determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS–PAGE). At the same time, its activity during the storage period was evaluated by gelatin zymography. Casein zymography was also performed to detect any caseinase contamination in the formulations. The release of the enzyme from different gel formulations was assessed using the Franz diffusion apparatus and analyzed by gelatin zymography. The results showed some physical changes that were more prominent in gels stored at room temperature than those kept refrigerated. The difference in humectant concentration was also found to affect the stability of gels. PG was found to be a better humectant than GL, particularly in a concentration of 25%. The zymography results indicated that collagenase was stable in all formulations kept in the refrigerator. In contrast, its complete degradation was noted in the preparations stored at room temperature within a month. The data generated in this study will help the formulators to commercialize a relatively economical gel formulation of collagenase that is highly stable for up to six years at refrigerator temperature (5±3°C).</div></div>","PeriodicalId":16741,"journal":{"name":"Journal of pharmaceutical sciences","volume":"114 2","pages":"Pages 1264-1279"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143007134","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Formulation robustness study was performed for a biosimilar monoclonal antibody (IgG1) manufactured at Dr. Reddy's Laboratory, where the pH and concentration level of excipients in the drug product formulation were systematically varied from the target formulation. It was observed that the IgG1 formulation having relatively low pH and high citrate (buffer salt) concentration were predisposed to the formation of low molecular weight impurities. Mass spectrometry analysis of the mAb1 fragments detected the pyroglutamate species from LC-LC dimer and fragmentation in the –DKTH- amino acid sequence of the heavy chain. Blind docking indicated binding of citrate with Lysine 222 residue in the proximity of Cys224 could have potentially fragmented IgG1.
Dr. Reddy's实验室对一种生物仿制单克隆抗体（IgG1）进行了配方稳稳性研究，其中药物制剂制剂中的赋形剂的pH值和浓度水平与目标制剂有系统的变化。观察到，具有相对低pH值和高柠檬酸盐（缓冲盐）浓度的IgG1制剂易于形成低分子量杂质。MaB1片段的质谱分析发现LC-LC二聚体中存在焦谷氨酸，重链- dkth -氨基酸序列存在片段。盲对接表明，柠檬酸盐与Lysine 222残基在Cys224附近的结合可能导致IgG1断裂。
{"title":"Mechanism of low molecular weight impurity formation in an IgG1 monoclonal antibody formulation","authors":"Pinaki Basu , Nidhi Verma , Sigireddi Indra Kumar , Maya Nanath , Sireesha Goswamy Kaligatla , Giridhar Sivalanka , Veerabhadra Madurai Veeraraghavan , Lovisha Aggarwal , Sunil A Nankar , Ravi Kumar Marikanti , Murali Jayaraman","doi":"10.1016/j.xphs.2024.12.024","DOIUrl":"10.1016/j.xphs.2024.12.024","url":null,"abstract":"<div><div>Formulation robustness study was performed for a biosimilar monoclonal antibody (IgG1) manufactured at Dr. Reddy's Laboratory, where the pH and concentration level of excipients in the drug product formulation were systematically varied from the target formulation. It was observed that the IgG1 formulation having relatively low pH and high citrate (buffer salt) concentration were predisposed to the formation of low molecular weight impurities. Mass spectrometry analysis of the mAb1 fragments detected the pyroglutamate species from LC-LC dimer and fragmentation in the –DKTH- amino acid sequence of the heavy chain. Blind docking indicated binding of citrate with Lysine 222 residue in the proximity of Cys224 could have potentially fragmented IgG1.</div></div>","PeriodicalId":16741,"journal":{"name":"Journal of pharmaceutical sciences","volume":"114 2","pages":"Pages 1464-1471"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143007139","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.xphs.2025.01.011
Kirti Sawant , Rasha M. Elkanayati , Ahmed Almotairy , Michael A. Repka , Mashan Almutairi
Clotrimazole, an antifungal agent for treating vaginal candidiasis, faces challenges in localized delivery due to poor solubility, complexity of the vaginal environment, limited fluid for dissolution, and rapid self washout of the vagina. The study aimed to enhance clotrimazole solubility using hot-melt extrusion (HME) to develop vaginal films with adequate bioadhesion, mechanical strength, and extended-release properties. Different formulations were created by varying the ratios of polyethylene oxide (PEO) grades (N750 and N10) to adjust the films' properties. The films demonstrated extended-release profiles, prolonging clotrimazole release for up to eight hours, with a cumulative gradual and complete in- vitro release in 100 mL of simulated vaginal fluid with 0.5% sodium dodecyl sulfate. In contrast, the marketed vaginal ovules exhibited a rapid and complete release within 30 minutes of shell rupture. The release kinetics followed Krosmeyer-Peppas model, and zero-order release mechanism. Films containing 25% clotrimazole, 56.25% PEO N750, and 18.75% PEO N10 exhibited strength of 87.9 N, stiffness of 35 N/sec, and adhesive force of 3.85 N.mm. In conclusion, the novel clotrimazole-loaded vaginal films developed using HME technology enhanced the solubility and localized vaginal delivery of clotrimazole. The extended-release profile may reduce the dosing frequency, enhance patient adherence, and improve therapeutic outcomes.
{"title":"Clotrimazole mucoadhesive films with extended-release properties for vaginal candidiasis—A hot-melt extrusion application","authors":"Kirti Sawant , Rasha M. Elkanayati , Ahmed Almotairy , Michael A. Repka , Mashan Almutairi","doi":"10.1016/j.xphs.2025.01.011","DOIUrl":"10.1016/j.xphs.2025.01.011","url":null,"abstract":"<div><div>Clotrimazole, an antifungal agent for treating vaginal candidiasis, faces challenges in localized delivery due to poor solubility, complexity of the vaginal environment, limited fluid for dissolution, and rapid self washout of the vagina. The study aimed to enhance clotrimazole solubility using hot-melt extrusion (HME) to develop vaginal films with adequate bioadhesion, mechanical strength, and extended-release properties. Different formulations were created by varying the ratios of polyethylene oxide (PEO) grades (N750 and N10) to adjust the films' properties. The films demonstrated extended-release profiles, prolonging clotrimazole release for up to eight hours, with a cumulative gradual and complete <em>in- vitro</em> release in 100 mL of simulated vaginal fluid with 0.5% sodium dodecyl sulfate. In contrast, the marketed vaginal ovules exhibited a rapid and complete release within 30 minutes of shell rupture. The release kinetics followed Krosmeyer-Peppas model, and zero-order release mechanism. Films containing 25% clotrimazole, 56.25% PEO N750, and 18.75% PEO N10 exhibited strength of 87.9 N, stiffness of 35 N/sec, and adhesive force of 3.85 N.mm. In conclusion, the novel clotrimazole-loaded vaginal films developed using HME technology enhanced the solubility and localized vaginal delivery of clotrimazole. The extended-release profile may reduce the dosing frequency, enhance patient adherence, and improve therapeutic outcomes.</div></div>","PeriodicalId":16741,"journal":{"name":"Journal of pharmaceutical sciences","volume":"114 2","pages":"Pages 1296-1306"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143007284","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.xphs.2024.10.042
Negar Jafari , Justin T. Douglas , Sarah A. Neuenswander , Payam Kelich , Michael J. Hageman
Cyclodextrin complexation has a potential to modulate the physicochemical properties of peptide drugs. The ability of peptides to form an inclusion complex can be influenced by factors such as size, amino acid sequence of peptide, and the size and charge of the cyclodextrin cavity. In this study, the inclusion complexes of the cyclic peptide drug lanreotide acetate with two common β-cyclodextrin derivatives, Sulfobutyl ether β-CD (SBEβ-CD) and hydroxypropyl β-CD (HPβ-CD) were investigated. NMR spectroscopy was used to examine the interaction between β-cyclodextrin derivatives and specific residues of lanreotide. It was observed that the hydrophobic side chain of aromatic residues in the lanreotide sequence can fit into the cavities of both β-cyclodextrin derivatives. Additionally, NMR revealed a lower diffusion coefficient and higher hydrodynamic radius of complex, indicative of binding to the cavities. Each aromatic residue was individually studied by substituting alanine in lanreotide to measure its association binding with both β-cyclodextrin derivatives. The alanine-substitute study indicated a stronger binding of SBEβ-CD to Lanreotide compared to HPβ-CD. Docking studies suggested that the 1:1 inclusion complex is more favorable than higher-order complexes due to the steric hindrance and size considerations. Docking analysis indicated the stable conformation of all three aromatic side chains with both β-cyclodextrin derivatives, SBEβ-CD and HPβ-CD.
{"title":"β-Cyclodextrin derivatives bind aromatic side chains of the cyclic peptide lanreotide","authors":"Negar Jafari , Justin T. Douglas , Sarah A. Neuenswander , Payam Kelich , Michael J. Hageman","doi":"10.1016/j.xphs.2024.10.042","DOIUrl":"10.1016/j.xphs.2024.10.042","url":null,"abstract":"<div><div>Cyclodextrin complexation has a potential to modulate the physicochemical properties of peptide drugs. The ability of peptides to form an inclusion complex can be influenced by factors such as size, amino acid sequence of peptide, and the size and charge of the cyclodextrin cavity. In this study, the inclusion complexes of the cyclic peptide drug lanreotide acetate with two common β-cyclodextrin derivatives, Sulfobutyl ether β-CD (SBEβ-CD) and hydroxypropyl β-CD (HPβ-CD) were investigated. NMR spectroscopy was used to examine the interaction between β-cyclodextrin derivatives and specific residues of lanreotide. It was observed that the hydrophobic side chain of aromatic residues in the lanreotide sequence can fit into the cavities of both β-cyclodextrin derivatives. Additionally, NMR revealed a lower diffusion coefficient and higher hydrodynamic radius of complex, indicative of binding to the cavities. Each aromatic residue was individually studied by substituting alanine in lanreotide to measure its association binding with both β-cyclodextrin derivatives. The alanine-substitute study indicated a stronger binding of SBEβ-CD to Lanreotide compared to HPβ-CD. Docking studies suggested that the 1:1 inclusion complex is more favorable than higher-order complexes due to the steric hindrance and size considerations. Docking analysis indicated the stable conformation of all three aromatic side chains with both β-cyclodextrin derivatives, SBEβ-CD and HPβ-CD.</div></div>","PeriodicalId":16741,"journal":{"name":"Journal of pharmaceutical sciences","volume":"114 2","pages":"Pages 878-886"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142569096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.xphs.2024.10.038
Rong-Sheng Yang , Chengbei Li , Liliana Henriquez , Hongxia Wang , Jainik Panchal , Wendy Zhong , Hillary Schuessler
Polysorbate 80 (PS80), a widely used polymeric surfactant in biotherapeutic formulation, possesses a unique structural composition that effectively prevents protein aggregation in highly concentrated protein drug formulations. However, PS80 is susceptible to hydrolysis, due to the presence of fatty acid esters that can be enzymatically hydrolyzed, The unsaturated bonds in the fatty acids are prone to oxidative degradation when exposed to air, especially in the presence of transition metals such as iron and copper, which may be introduced during production and purification processes or from contamination in raw materials used in drug formulation. The degradation of PS80, particularly through metal-mediated oxidative degradation, poses a significant challenge for the industry. Among the identified trace metals, iron plays a crucial role as the redox reaction between ferrous ion (Fe(II)) and ferric ion (Fe(III)) generates radicals that initiate the degradation process. In order to investigate the impact of iron on PS80 degradation and understand the mechanism of iron-catalyzed oxidation, we utilized charge-reduction mass spectrometry and two-dimensional ion density mapping technologies to characterize the degradation of PS80. This method has proven to be a convenient and effective tool for the quick and detailed profiling of PS80, allowing for visual monitoring and examination of the changes that reflect the difficult-to-identify and easy-to-miss oxidized species of PS80. Additionally, a high-performance liquid chromatography coupled to inductively coupled plasma mass spectrometry method was developed for the separation and measurement of Fe(II) and Fe(III). Through this investigation, we determined that the involvement of Fe(II)/Fe(III) in PS80 degradation is a temperature dependent process. Furthermore, we found citrate not only promotes the conversion of Fe(II) to Fe(III), but it also chelates Fe(III) and prevents its reduction to Fe(II), thus inhibiting the initiation of the PS80 degradation. Therefore, the addition of citrate can be a crucial ingredient for controlling the degradation of PS80 in biologic drug substances and products. Overall, this investigation has provided valuable insights to enhance product stability, optimize processes, and ensure the quality of formulations containing PS80.
{"title":"Impact of citrate on mitigating iron mediated polysorbate 80 degradation in biotherapeutic formulation placebos","authors":"Rong-Sheng Yang , Chengbei Li , Liliana Henriquez , Hongxia Wang , Jainik Panchal , Wendy Zhong , Hillary Schuessler","doi":"10.1016/j.xphs.2024.10.038","DOIUrl":"10.1016/j.xphs.2024.10.038","url":null,"abstract":"<div><div>Polysorbate 80 (PS80), a widely used polymeric surfactant in biotherapeutic formulation, possesses a unique structural composition that effectively prevents protein aggregation in highly concentrated protein drug formulations. However, PS80 is susceptible to hydrolysis, due to the presence of fatty acid esters that can be enzymatically hydrolyzed, The unsaturated bonds in the fatty acids are prone to oxidative degradation when exposed to air, especially in the presence of transition metals such as iron and copper, which may be introduced during production and purification processes or from contamination in raw materials used in drug formulation. The degradation of PS80, particularly through metal-mediated oxidative degradation, poses a significant challenge for the industry. Among the identified trace metals, iron plays a crucial role as the redox reaction between ferrous ion (Fe(II)) and ferric ion (Fe(III)) generates radicals that initiate the degradation process. In order to investigate the impact of iron on PS80 degradation and understand the mechanism of iron-catalyzed oxidation, we utilized charge-reduction mass spectrometry and two-dimensional ion density mapping technologies to characterize the degradation of PS80. This method has proven to be a convenient and effective tool for the quick and detailed profiling of PS80, allowing for visual monitoring and examination of the changes that reflect the difficult-to-identify and easy-to-miss oxidized species of PS80. Additionally, a high-performance liquid chromatography coupled to inductively coupled plasma mass spectrometry method was developed for the separation and measurement of Fe(II) and Fe(III). Through this investigation, we determined that the involvement of Fe(II)/Fe(III) in PS80 degradation is a temperature dependent process. Furthermore, we found citrate not only promotes the conversion of Fe(II) to Fe(III), but it also chelates Fe(III) and prevents its reduction to Fe(II), thus inhibiting the initiation of the PS80 degradation. Therefore, the addition of citrate can be a crucial ingredient for controlling the degradation of PS80 in biologic drug substances and products. Overall, this investigation has provided valuable insights to enhance product stability, optimize processes, and ensure the quality of formulations containing PS80.</div></div>","PeriodicalId":16741,"journal":{"name":"Journal of pharmaceutical sciences","volume":"114 2","pages":"Pages 857-865"},"PeriodicalIF":3.7,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142639192","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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