URL: https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631 TITLE: Effectiveness of Lyoprotectants in Protein Stabilization During ... ERROR: None [Skip to main content](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#main-content) ![Image 1](https://cdn.ncbi.nlm.nih.gov/pmc/pd-medc-pmc-cloudpmc-viewer/production/a2b04810/var/data/static/img/us_flag.svg) An official website of the United States government Here's how you know Here's how you know ![Image 2](https://cdn.ncbi.nlm.nih.gov/pmc/pd-medc-pmc-cloudpmc-viewer/production/a2b04810/var/data/static/img/icon-dot-gov.svg) **Official websites use .gov** A **.gov** website belongs to an official government organization in the United States. ![Image 3](https://cdn.ncbi.nlm.nih.gov/pmc/pd-medc-pmc-cloudpmc-viewer/production/a2b04810/var/data/static/img/icon-https.svg) **Secure .gov websites use HTTPS** A **lock** ( ) or **https://** means you've safely connected to the .gov website. 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Learn more: [PMC Disclaimer](https://pmc.ncbi.nlm.nih.gov/about/disclaimer/) | [PMC Copyright Notice](https://pmc.ncbi.nlm.nih.gov/about/copyright/) ![Image 14: Pharmaceutics logo](https://cdn.ncbi.nlm.nih.gov/pmc/banners/logo-pharmamdpi.png) Pharmaceutics . 2024 Oct 21;16(10):1346. doi: [10.3390/pharmaceutics16101346](https://doi.org/10.3390/pharmaceutics16101346) * [Search in PMC](https://pmc.ncbi.nlm.nih.gov/search/?term=%22Pharmaceutics%22[jour]) * [Search in PubMed](https://pubmed.ncbi.nlm.nih.gov/?term=%22Pharmaceutics%22[jour]) * [View in NLM Catalog](https://www.ncbi.nlm.nih.gov/nlmcatalog?term=%22Pharmaceutics%22[Title%20Abbreviation]) * [Add to search](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631?term=%22Pharmaceutics%22[jour]) # Effectiveness of Lyoprotectants in Protein Stabilization During Lyophilization [Vinoothini Karunnanithy](https://pubmed.ncbi.nlm.nih.gov/?term=%22Karunnanithy%20V%22[Author]) ### Vinoothini Karunnanithy 1 Department of Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia; vinoothini8205@gmail.com (V.K.); atiqah.haizum@ukm.edu.my (N.A.H.A.); fauzibusra@ukm.edu.my (M.B.F.); lyoges@ppukm.ukm.edu.my (Y.L.); angela@ppukm.ukm.edu.my (A.N.M.H.) Find articles by [Vinoothini Karunnanithy](https://pubmed.ncbi.nlm.nih.gov/?term=%22Karunnanithy%20V%22[Author]) 1, [Nur Hazirah Binti Abdul Rahman](https://pubmed.ncbi.nlm.nih.gov/?term=%22Abdul%20Rahman%20N%22[Author]) ### Nur Hazirah Binti Abdul Rahman 1 Department of Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia; vinoothini8205@gmail.com (V.K.); atiqah.haizum@ukm.edu.my (N.A.H.A.); fauzibusra@ukm.edu.my (M.B.F.); lyoges@ppukm.ukm.edu.my (Y.L.); angela@ppukm.ukm.edu.my (A.N.M.H.) Find articles by [Nur Hazirah Binti Abdul Rahman](https://pubmed.ncbi.nlm.nih.gov/?term=%22Abdul%20Rahman%20N%22[Author]) 1, [Nur Atiqah Haizum Abdullah](https://pubmed.ncbi.nlm.nih.gov/?term=%22Abdullah%20N%22[Author]) ### Nur Atiqah Haizum Abdullah 1 Department of Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia; vinoothini8205@gmail.com (V.K.); atiqah.haizum@ukm.edu.my (N.A.H.A.); fauzibusra@ukm.edu.my (M.B.F.); lyoges@ppukm.ukm.edu.my (Y.L.); angela@ppukm.ukm.edu.my (A.N.M.H.) Find articles by [Nur Atiqah Haizum Abdullah](https://pubmed.ncbi.nlm.nih.gov/?term=%22Abdullah%20N%22[Author]) 1, [Mh Busra Fauzi](https://pubmed.ncbi.nlm.nih.gov/?term=%22Fauzi%20M%22[Author]) ### Mh Busra Fauzi 1 Department of Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia; vinoothini8205@gmail.com (V.K.); atiqah.haizum@ukm.edu.my (N.A.H.A.); fauzibusra@ukm.edu.my (M.B.F.); lyoges@ppukm.ukm.edu.my (Y.L.); angela@ppukm.ukm.edu.my (A.N.M.H.) 2 Advance Bioactive Materials-Cells UKM Research Group, Universiti Kebangsaan Malaysia, Bangi 43600, Selangor, Malaysia Find articles by [Mh Busra Fauzi](https://pubmed.ncbi.nlm.nih.gov/?term=%22Fauzi%20M%22[Author]) 1 2, [Yogeswaran Lokanathan](https://pubmed.ncbi.nlm.nih.gov/?term=%22Lokanathan%20Y%22[Author]) ### Yogeswaran Lokanathan 1 Department of Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia; vinoothini8205@gmail.com (V.K.); atiqah.haizum@ukm.edu.my (N.A.H.A.); fauzibusra@ukm.edu.my (M.B.F.); lyoges@ppukm.ukm.edu.my (Y.L.); angela@ppukm.ukm.edu.my (A.N.M.H.) 2 Advance Bioactive Materials-Cells UKM Research Group, Universiti Kebangsaan Malaysia, Bangi 43600, Selangor, Malaysia Find articles by [Yogeswaran Lokanathan](https://pubmed.ncbi.nlm.nih.gov/?term=%22Lokanathan%20Y%22[Author]) 1 2, [Angela Ng Min Hwei](https://pubmed.ncbi.nlm.nih.gov/?term=%22Min%20Hwei%20A%22[Author]) ### Angela Ng Min Hwei 1 Department of Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia; vinoothini8205@gmail.com (V.K.); atiqah.haizum@ukm.edu.my (N.A.H.A.); fauzibusra@ukm.edu.my (M.B.F.); lyoges@ppukm.ukm.edu.my (Y.L.); angela@ppukm.ukm.edu.my (A.N.M.H.) 2 Advance Bioactive Materials-Cells UKM Research Group, Universiti Kebangsaan Malaysia, Bangi 43600, Selangor, Malaysia Find articles by [Angela Ng Min Hwei](https://pubmed.ncbi.nlm.nih.gov/?term=%22Min%20Hwei%20A%22[Author]) 1 2, [Manira Maarof](https://pubmed.ncbi.nlm.nih.gov/?term=%22Maarof%20M%22[Author]) ### Manira Maarof 1 Department of Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia; vinoothini8205@gmail.com (V.K.); atiqah.haizum@ukm.edu.my (N.A.H.A.); fauzibusra@ukm.edu.my (M.B.F.); lyoges@ppukm.ukm.edu.my (Y.L.); angela@ppukm.ukm.edu.my (A.N.M.H.) 2 Advance Bioactive Materials-Cells UKM Research Group, Universiti Kebangsaan Malaysia, Bangi 43600, Selangor, Malaysia 3 Ageing and Degenerative Disease UKM Research Group, Universiti Kebangsaan Malaysia, Bangi 43600, Selangor, Malaysia Find articles by [Manira Maarof](https://pubmed.ncbi.nlm.nih.gov/?term=%22Maarof%20M%22[Author]) 1 2 3* Editor: Kyriakos Kachrimanis * Author information * Article notes * Copyright and License information 1 Department of Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia; vinoothini8205@gmail.com (V.K.); atiqah.haizum@ukm.edu.my (N.A.H.A.); fauzibusra@ukm.edu.my (M.B.F.); lyoges@ppukm.ukm.edu.my (Y.L.); angela@ppukm.ukm.edu.my (A.N.M.H.) 2 Advance Bioactive Materials-Cells UKM Research Group, Universiti Kebangsaan Malaysia, Bangi 43600, Selangor, Malaysia 3 Ageing and Degenerative Disease UKM Research Group, Universiti Kebangsaan Malaysia, Bangi 43600, Selangor, Malaysia ✉ *Correspondence: manira@ppukm.ukm.edu.my; Tel.: +60-603-9145-8226 #### Roles **Kyriakos Kachrimanis**: Academic Editor Received 2024 Sep 11; Revised 2024 Oct 10; Accepted 2024 Oct 17; Collection date 2024. © 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ([https://creativecommons.org/licenses/by/4.0/](https://creativecommons.org/licenses/by/4.0/)). [PMC Copyright notice](https://pmc.ncbi.nlm.nih.gov/about/copyright/) PMCID: PMC11510631 PMID: [39458674](https://pubmed.ncbi.nlm.nih.gov/39458674/) ## [](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631)Abstract **Background:** Proteins are commonly used in the healthcare industry to treat various health conditions, and most proteins are sensitive to physical and chemical changes. Lyophilization, also known as freeze-drying, involves sublimating water in the form of ice from a substance at low pressure, forming a freeze-dried powder that increases its shelf life. Extreme pressure and varying temperatures in the freeze-drying process may damage the protein’s structural integrity. Lyoprotectants are commonly used to protect protein conformations. It is important to choose a suitable lyoprotectant to ensure optimal effectiveness. **Method:** Twenty articles screened from Scopus, Web of Science, and PubMed were included in this review that discussed potential lyoprotectants and their effectiveness with different protein models. **Results:** Lyoprotectants were categorized into sugars, polyols, surfactants, and amino acids. Lyoprotectants can exhibit significant protective effects towards proteins, either singularly or in combination with another lyoprotectant. They exert various interactions with the protein to stabilize it, such as hydrogen bonding, hydrophobic interactions, electrostatic interactions, and osmoprotection. **Conclusions:** This review concludes that disaccharides are the most effective lyoprotectants, while other groups of lyoprotectants are best used in combination with other lyoprotectants. **Keywords:**lyoprotectant, protein stabilization, freeze-drying, cryodesiccation ## [](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631)1. Introduction Freeze-drying, also known as lyophilization, is widely used to preserve proteins in a dry state for long-term storage and transportation. These frozen-dried bioproducts facilitate transportation to distant locations without the need for cold-chain storage and reduce point-of-care labor costs by eliminating the need for on-site mixing of reagents. Lyophilized protein powder can be stocked for future outbreaks, and this strategy can even be used in experiments with more extended time scales [[1](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B1-pharmaceutics-16-01346),[2](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B2-pharmaceutics-16-01346)]. Lyophilization is an ideal process to convert protein solution into a solid form. The freeze-dried protein powders will increase their shelf-life. However, protein integrity is compromised since the drying process exerts pressure and causes irreversible conformational changes in the protein structure, which leads to the inactivation of the protein functions. During reconstitution of the freeze-dried protein powder, moisture causes the freeze-dried proteins to undergo disulphide interchange and other reactions, which lead to inactivation [[1](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B1-pharmaceutics-16-01346),[2](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B2-pharmaceutics-16-01346)]. A FTIR spectroscopy study conducted by Ipsita Roy and team in 2004 exhibited that freeze-drying proteins leads to conformational changes in protein structure. They concluded that the drying stage decreased the α-helix and random structure and increased the β-sheet form in the protein. Freeze-dried fibroblast growth factor and γ-interferon proteins showed significant conformational changes and aggregation during freeze-drying [[3](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B3-pharmaceutics-16-01346)]. In order to prevent denaturation and aggregation of protein molecules, lyoprotectants need to be added to stabilize the protein conformation. When aqueous phase stability is a barrier, lyophilization is the most popular method to generate a freeze-dried product. It is crucial to preserve materials that require a sterile and delicate preservation technique and minimal moisture (less than 1%) to maintain stability [[4](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B4-pharmaceutics-16-01346),[5](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B5-pharmaceutics-16-01346)]. Freeze-drying is primarily used to safely eliminate moisture from sensitive products, typically those with biological origins, so they can be stored easily, kept in a state suitable for long-term storage, and reactivated by just adding water [[1](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B1-pharmaceutics-16-01346),[3](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B3-pharmaceutics-16-01346)]. However, additional stressors from the freeze-drying procedure may result in protein aggregation, fragmentation, and loss of activity. The process usually leads to an increase in the β-sheet formation and a decrease in the α-helix and random structure [[3](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B3-pharmaceutics-16-01346)]. It is commonly known that cryoprotectants can protect the protein from denaturation in the early phases, while lyoprotectants are necessary to prevent protein inactivation after drying [[6](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B6-pharmaceutics-16-01346),[7](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B7-pharmaceutics-16-01346)]. Adding cryoprotectants alone is not sufficient to ensure protein integrity, as the drying phase can significantly impact protein stability. Therefore, lyoprotectants are required to shield the proteins during the drying phase [[3](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B3-pharmaceutics-16-01346),[8](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B8-pharmaceutics-16-01346)]. Lyoprotectants are added to protein formulations to protect them from freeze-drying stresses. They achieve this by reducing the water content and maintaining proper protein structure and stability during the process. The choice of lyoprotectant depends on the protein characteristics, freezing conditions, and desired final product properties. Commonly used lyoprotectants include sugars, polyols, amino acids, and surfactants [[9](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B9-pharmaceutics-16-01346)]. Choosing the right lyoprotectant is crucial to preserving the stability and activity of proteins during the freeze-drying process. In protein formulations, sugars, polyols, amino acids, and surfactants are often utilized as lyoprotectants. To create novel and more potent lyoprotectants for protein stabilization during freeze-drying, more investigation is required. This systematic review discusses and identifies the most promising lyoprotectant to preserve protein structure during freeze-drying. ## [](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631)2. Methodology ### 2.1. Search Strategy A comprehensive search was performed in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The search was conducted in electronic databases such as PubMed, Web of Science (WOS), and Scopus. To ensure only current studies were used, the search was restricted to publications from 2018 to 2024. Three sets of keyword terms were used for the searching process: “lyoprotectants” or “protective agents” or “chemical additives”; “protein stabilization” or “conformational stability” or “protein activity”; and” lyophilization” or “freeze-drying” or “cryodesiccation”. The title and abstract of all articles were screened individually and duplicate articles were excluded. The references of all retrieved and relevant articles were also reviewed to ensure more thorough findings. The search was restricted to published articles and journal clinical trials in English language. [Figure 1](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#pharmaceutics-16-01346-f001) shows the comprehensive search results which were performed according to PRISMA guidelines. The details of search strategy and PRISMA checklist are recorded in the [Supplementary Tables](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#app1-pharmaceutics-16-01346) [[10](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#B10-pharmaceutics-16-01346)]. ### 2.2. Data Extraction and Synthesis Two authors (V.K. and N.A.H.A.) independently recorded data from every included study. Any disagreement between the two authors was resolved by discussion with other authors. Then, the following data were extracted: study design, type lyoprotectants, effect on protein models, and alternative freeze-drying method. The inclusion and exclusion criteria outlined are essential for determining the scope of the systematic review focused on lyoprotectants for protein stabilization during freeze-drying. The review considered studies published between 2018 and 2024. This time frame ensures that the review was based on the most recent and relevant research, reflecting the latest advancements in the field. Only full-text articles published in peer-reviewed scientific journals were included. This criterion was crucial for ensuring the credibility and scientific rigor of the studies being reviewed. This review included articles published in English. This limitation was set to facilitate comprehensive understanding and accessibility of the research findings. This review encompassed studies conducted globally, allowing for a wide range of data and insights from various geographical contexts. Only reviews of published original research articles in indexed journals were included. This criterion ensured that the review was grounded in established scientific evidence rather than anecdotal or unverified claims. For the exclusion criteria, articles that were not present in indexed journals, such as online articles, review articles, letters to editors, perspectives, commentaries, and news reports, were excluded. This helped maintain the quality and reliability of the sources included in the review. Articles published in languages other than English were not considered. This exclusion was necessary due to the challenges of accurately interpreting and translating scientific content, which could lead to misinterpretation of the findings. By adhering to these inclusion and exclusion criteria, the systematic review aimed to compile a comprehensive and credible body of evidence regarding the effectiveness of lyoprotectants for protein stabilization during the freeze-drying process. ## [](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631)3. Results and Discussion This systematic review was aimed to screen and identify potential lyoprotectants. Discovering new lyoprotectants is important for several reasons, primarily related to enhancing the effectiveness, stability, and safety of the lyophilization (freeze-drying) process, which is widely used for preserving sensitive biological materials, pharmaceuticals, and food products. As formulations become more complex, containing multiple active ingredients or excipients, discovering new lyoprotectants that are compatible with these formulations becomes crucial. Effective lyoprotectants facilitate faster and more complete reconstitution of the lyophilized product back into solution. This is particularly important in clinical settings, where quick preparation is often necessary. As new classes of therapeutics, such as mRNA-based drugs and gene therapies, are developed, they may require unique lyoprotectants to maintain their stability and effectiveness. The database search yielded 79 results in total. Out of these, 42 articles failed to match the inclusion criteria, and 17 were discarded as duplicates. Ultimately, 20 studies were included in this review. We discovered plausible lyoprotectants from multiple sources that can preserve various protein models during the freeze-drying process after reviewing and contrasting our databases. According to the data, we list the most commonly used lyoprotectant, new potential protein stabilizers, effectiveness of combining two or more lyoprotectants, and interactions between the lyoprotectants and protein. [Table 1](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#pharmaceutics-16-01346-t001) summarizes the identified lyoprotectants from the selected articles for further analysis. ### 3.1. Classification of Lyoprotectants [... middle omitted — see footer ...] * 58..Piszkiewicz S., Pielak G.J.. 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[[Google Scholar](https://scholar.google.com/scholar_lookup?journal=Pharm.%20Res.&title=Advances%20in%20the%20development%20of%20lyophilized%20formulations%20for%20monoclonal%20antibodies&author=Shalaeva%20Y.V.&author=Izmailov%20S.F.&volume=27&publication_year=2010&pages=393-400&)] # [](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631) Associated Data _This section collects any data citations, data availability statements, or supplementary materials included in this article._ ## Supplementary Materials [pharmaceutics-16-01346-s001.zip](https://pmc.ncbi.nlm.nih.gov/articles/instance/11510631/bin/pharmaceutics-16-01346-s001.zip) * * * Articles from Pharmaceutics are provided here courtesy of **Multidisciplinary Digital Publishing Institute (MDPI)** ![Image 15: Close](https://cdn.ncbi.nlm.nih.gov/pmc/pd-medc-pmc-cloudpmc-viewer/production/a2b04810/var/data/static/img/usa-icons/close.svg) ## ACTIONS * [![Image 16: View on publisher site icon](https://cdn.ncbi.nlm.nih.gov/pmc/pd-medc-pmc-cloudpmc-viewer/production/a2b04810/var/data/static/img/launch.svg)View on publisher site](https://doi.org/10.3390/pharmaceutics16101346) * [![Image 17: Download PDF icon](https://cdn.ncbi.nlm.nih.gov/pmc/pd-medc-pmc-cloudpmc-viewer/production/a2b04810/var/data/static/img/file_download.svg)PDF](https://pmc.ncbi.nlm.nih.gov/articles/pdf/pharmaceutics-16-01346.pdf) * ![Image 18: Cite icon](https://cdn.ncbi.nlm.nih.gov/pmc/pd-medc-pmc-cloudpmc-viewer/production/a2b04810/var/data/static/img/format_quote.svg)Cite * ![Image 19: Collections icon](https://cdn.ncbi.nlm.nih.gov/pmc/pd-medc-pmc-cloudpmc-viewer/production/a2b04810/var/data/static/img/action-bookmark-full.svg)![Image 20: Collections icon](https://cdn.ncbi.nlm.nih.gov/pmc/pd-medc-pmc-cloudpmc-viewer/production/a2b04810/var/data/static/img/action-bookmark-empty.svg)Collections * ![Image 21: Permalink icon](https://cdn.ncbi.nlm.nih.gov/pmc/pd-medc-pmc-cloudpmc-viewer/production/a2b04810/var/data/static/img/share.svg)Permalink## PERMALINK ![Image 22: Copy icon](https://cdn.ncbi.nlm.nih.gov/pmc/pd-medc-pmc-cloudpmc-viewer/production/a2b04810/var/data/static/img/content_copy.svg)Copy ## RESOURCES ### Similar articles ### Cited by other articles ### Links to NCBI Databases ## On this page * [Abstract](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#abstract1) * [1. Introduction](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#sec1-pharmaceutics-16-01346) * [2. Methodology](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#sec2-pharmaceutics-16-01346) * [3. Results and Discussion](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#sec3-pharmaceutics-16-01346) * [4. Limitation and Challenges](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#sec4-pharmaceutics-16-01346) * [5. Conclusions](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#sec5-pharmaceutics-16-01346) * [Funding Statement](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#funding-statement1) * [Acknowledgments](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#ack) * [Supplementary Materials](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#app1-pharmaceutics-16-01346) * [References](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#ref-list1) * [Associated Data](https://pmc.ncbi.nlm.nih.gov/articles/PMC11510631#_ad93_) Follow NCBI [NCBI on X (formerly known as Twitter)](https://twitter.com/ncbi)[NCBI on Facebook](https://www.facebook.com/ncbi.nlm)[NCBI on LinkedIn](https://www.linkedin.com/company/ncbinlm)[NCBI on GitHub](https://github.com/ncbi)[NCBI RSS feed](https://ncbiinsights.ncbi.nlm.nih.gov/) Connect with NLM [NLM on X (formerly known as Twitter)](https://twitter.com/nlm_nih)[NLM on Facebook](https://www.facebook.com/nationallibraryofmedicine)[NLM on YouTube](https://www.youtube.com/user/NLMNIH) [National Library of Medicine 8600 Rockville Pike Bethesda, MD 20894](https://www.google.com/maps/place/8600+Rockville+Pike,+Bethesda,+MD+20894/%4038.9959508,%20%20%20%20%20%20%20%20%20%20%20%20-77.101021,17z/data%3D!3m1!4b1!4m5!3m4!1s0x89b7c95e25765ddb%3A0x19156f88b27635b8!8m2!3d38.9959508!%20%20%20%20%20%20%20%20%20%20%20%204d-77.0988323) * [Web Policies](https://www.nlm.nih.gov/web_policies.html) * [FOIA](https://www.nih.gov/institutes-nih/nih-office-director/office-communications-public-liaison/freedom-information-act-office) * [HHS Vulnerability Disclosure](https://www.hhs.gov/vulnerability-disclosure-policy/index.html) * [Help](https://support.nlm.nih.gov/?pagename=cloudpmc-viewer%3Apmc%3Aarticle%3Ajournal) * [Accessibility](https://www.nlm.nih.gov/accessibility.html) * [Careers](https://www.nlm.nih.gov/careers/careers.html) * [NLM](https://www.nlm.nih.gov/) * [NIH](https://www.nih.gov/) * [HHS](https://www.hhs.gov/) * [USA.gov](https://www.usa.gov/) Back to Top ![Image 23: back to top icon](https://cdn.ncbi.nlm.nih.gov/pmc/pd-medc-pmc-cloudpmc-viewer/production/a2b04810/var/data/static/img/arrow_upward.svg) ![Image 24: Tell us what you think!](https://iad1.qualtrics.com/WRQualtricsSiteIntercept/Graphic.php?IM=IM_3xWqwNkIHc7cA0S) ──────── [TRUNCATED] ──────── Showing 22,037 chars (head) + 6,896 chars (tail) of 112,575 total clean characters. 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