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Photoacoustic imaging reveals mechanisms of rapid-acting insulin formulations dynamics at the injection site

Khadria, Anjul and Paavola, Chad D. and Maslov, Konstantin and Valenzuela, Francisco A. and Sperry, Andrea E. and Cox, Amy L. and Cao, Rui and Shi, Junhui and Brown-Augsburger, Patricia L. and Lozano, Emmanuel and Blankenship, Ross L. and Majumdar, Ranajoy and Bradley, Scott A. and Beals, John M. and Oladipupo, Sunday S. and Wang, Lihong V. (2022) Photoacoustic imaging reveals mechanisms of rapid-acting insulin formulations dynamics at the injection site. Molecular Metabolism, 62 . Art. No. 101522. ISSN 2212-8778. PMCID PMC9207296. doi:10.1016/j.molmet.2022.101522. https://resolver.caltech.edu/CaltechAUTHORS:20220315-626283000

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Abstract

Objective: Ultra-rapid insulin formulations control postprandial hyperglycemia; however, inadequate understanding of injection site absorption mechanisms is limiting further advancement. We used photoacoustic imaging to investigate the injection site dynamics of dye-labeled insulin lispro in the Humalog® and Lyumjev® formulations using the murine ear cutaneous model and correlated it with results from unlabeled insulin lispro in pig subcutaneous injection model. Methods: We employed dual-wavelength optical-resolution photoacoustic microscopy to study the absorption and diffusion of the near-infrared dye-labeled insulin lispro in the Humalog and Lyumjev formulations in mouse ears. We mathematically modeled the experimental data to calculate the absorption rate constants and diffusion coefficients. We studied the pharmacokinetics of the unlabeled insulin lispro in both the Humalog and Lyumjev formulations as well as a formulation lacking both the zinc and phenolic preservative in pigs. The association state of insulin lispro in each of the formulations was characterized using SV-AUC and NMR spectroscopy. Results: Through experiments using murine and swine models, we show that the hexamer dissociation rate of insulin lispro is not the absorption rate-limiting step. We demonstrated that the excipients in the Lyumjev formulation produce local tissue expansion and speed both insulin diffusion and microvascular absorption. We also show that the diffusion of insulin lispro at the injection site drives its initial absorption; however, the rate at which the insulin lispro crosses the blood vessels is its overall absorption rate-limiting step. Conclusions: This study provides insights into injection site dynamics of insulin lispro and the impact of formulation excipients. It also demonstrates photoacoustic microscopy as a promising tool for studying protein therapeutics. The results from this study address critical questions around the subcutaneous behavior of insulin lispro and the formulation excipients, which could be useful to make faster and better controlled insulin formulations in the future.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.molmet.2022.101522DOIArticle
https://ars.els-cdn.com/content/image/1-s2.0-S2212877822000916-mmc1.zipPublisherSupplementary data
http://www.ncbi.nlm.nih.gov/pmc/articles/pmc9207296/PubMed CentralArticle
https://doi.org/10.1101/2022.03.10.483309DOIDiscussion Paper
ORCID:
AuthorORCID
Khadria, Anjul0000-0002-9771-3650
Maslov, Konstantin0000-0003-3408-8840
Cao, Rui0000-0003-4444-7528
Shi, Junhui0000-0002-5741-2781
Oladipupo, Sunday S.0000-0001-6555-0693
Wang, Lihong V.0000-0001-9783-4383
Additional Information:© 2022 The Author(s). Published by Elsevier GmbH. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). Received 10 March 2022, Revised 20 May 2022, Accepted 24 May 2022, Available online 4 June 2022, Version of Record 14 June 2022. We thank Steven Cottle and Linh Nguyen, Eli Lilly and Company for preparing the insulin formulation solutions. We also thank Jake Anderson, Eli Lilly and Company for useful discussions. This work was funded by Eli Lilly and Company. Data availability: The data that support the conclusions are present in the main draft or the supplementary information. The data processing and quantification procedures are described in detail in the Methods section. Author contributions: L.V.W., S.S.O., J.M.B., C.D.P., and A.K. conceived the project and the ideas. C.D.P. designed the chemistry for dye labeling and analyzed the chemistry data. F.A.V. labeled the insulin molecules with the sulfo-cyanine7.5 dye and characterized them. F.A.V. and A.K. prepared the insulin and dye buffer solutions. A.K. and K.M. designed and built the scanning photoacoustic microscope. J.S. and R.C. wrote the LabVIEW software for photoacoustic data acquisition. A.K. designed and performed all the photoacoustic experiments, wrote the MATLAB codes, and analyzed all the photoacoustic data. A.K., K.M., and L.V.W. designed the photoacoustic quantification algorithm. A.K. and K.M. devised the pharmacokinetics and diffusion methodologies. A.L.C. designed and supported the pharmacokinetics study in pigs and A.E.S. performed the pharmacokinetic experiments. R.M. prepared the unlabeled insulin lispro samples, and designed and performed the SV-AUC experiments. S.A.B. designed and performed the NMR spectroscopy analyses. A.K., C.D.P., K.M., J.M.B, and L.V.W. interpreted the final data. P.B.A., E.L., and R.L.B. were involved in active discussions. L.V.W., S.S.O., and J.M.B. supervised the project. A.K. wrote the manuscript. C.D.P., K.M., R.C., J.M.B, S.S.O., and L.V.W. contributed to writing the manuscript. Competing interests: A.K., R.C., and J.S. declare no competing interests. C.D.P., F.A.V, A.E.S., A.M.C., P.L.B.A., E.L., R.L.B, R.M., SA.B., J.M.B, and S.S.O. are employees and stockholders of Eli Lilly and Company. L.V.W. and K.M. have financial interests in Microphotoacoustics, Inc., CalPACT, LLC and Union Photoacoustic Technologies, Ltd, which did not support this work. L.V.W. received a contract from Eli Lilly and Company to conduct the studies reported herein.
Funders:
Funding AgencyGrant Number
Eli Lilly and CompanyUNSPECIFIED
Subject Keywords:Insulin; Photoacoustic imaging; Pharmacokinetic modeling; Diffusion; Diabetes; In vivo imaging
PubMed Central ID:PMC9207296
DOI:10.1016/j.molmet.2022.101522
Record Number:CaltechAUTHORS:20220315-626283000
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20220315-626283000
Official Citation:Anjul Khadria, Chad D. Paavola, Konstantin Maslov, Francisco A. Valenzuela, Andrea E. Sperry, Amy L. Cox, Rui Cao, Junhui Shi, Patricia L. Brown-Augsburger, Emmanuel Lozano, Ross L. Blankenship, Ranajoy Majumdar, Scott A. Bradley, John M. Beals, Sunday S. Oladipupo, Lihong V. Wang, Photoacoustic imaging reveals mechanisms of rapid-acting insulin formulations dynamics at the injection site, Molecular Metabolism, Volume 62, 2022, 101522, ISSN 2212-8778, https://doi.org/10.1016/j.molmet.2022.101522.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:113921
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:16 Mar 2022 14:35
Last Modified:28 Jun 2022 18:15

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