Impurities in Oligonucleotide Drug Substances and Drug Products

Capaldi, Daniel C.; Teasdale, Andy; Henry, Scott P.; Akhtar, Nadim; Besten, C. den; Gao-Sheridan, Samantha; Kretschmer, Matthias; Sharpe, Neal; Andrews, B. J.; Burm, Brigitte E. A.; Foy, Jeffrey W.-D.

doi:10.1089/nat.2017.0691

Cited by 88 publications

(75 citation statements)

References 44 publications

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“…Native impurity levels in a representative lot of ISIS 304801 range from 0.26% to 2.5% of total full-length oligonucleotide. The categorization of impurities followed the principles put forward in [1]. In short, the phosphodiester, n -1, and n + 1 impurities were not considered to need any safety characterization because the impurities were either similar to native DNA, consistent with common metabolites, or structurally similar to the parent oligonucleotide.…”

Section: Discussionmentioning

confidence: 99%

“…The enriched impurities are noted in bold font in Table 1. The test article formulations were prepared by combining oligonucleotide materials with elevated levels of specific impurities, which were in turn manufactured using two techniques [1]. Impurities that could be made by incorporation of a modified phosphoramidite during synthesis [ie 2¢-O-CH 3 , 2¢-O-butyl, 2¢-O-(2-ethoxyethyl), CNET, Dithioate, 3¢-MOE, Inverted, Early Eluting Impurities, and Late Eluting Impurities] were generated by synthesizing ISIS 304801 with the appropriate mixture of the corresponding phosphoramidite impurities and the parent nucleoside phosphoramidites.…”

Section: Methodsmentioning

confidence: 99%

“…A s therapeutic oligonucleotides progress through the nonclinical and clinical development process, one important element is to evaluate the safety considerations and exposure coverage for potential drug impurities. Recent efforts by a diverse group of industry scientists published broad principles that inform the characterization and classification of oligonucleotide-related impurities [1]. The goal of this publication was to describe the chemical context of impurities for this class of drugs and establish ''best practices'' specific to oligonucleotides with the purpose of aligning processes, as relatively few programs have advanced to the review and approval stage.…”

Section: Introductionmentioning

confidence: 99%

“…The other implication is that impurities resulting from starting materials or the process may be incorporated at different positions within the oligonucleotide. A more detailed catalog of product-related oligonucleotide impurities by broad structural types is presented by Capaldi et al [1].…”

Section: Introductionmentioning

confidence: 99%

“…An article outlining principles in the safety assessment of oligonucleotide-related impurities, taking into consideration the chemical nature of the impurities and the impurity levels, was published recently [1]. This article includes categorization of certain classes of impurities to guide testing paradigms, as well as recommendations about how best to apply existing guidelines for testing of oligonucleotides.…”

Section: Introductionmentioning

confidence: 99%

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Impurity Qualification Toxicology Study for a 2′-O-Methoxyethyl-Modified Antisense Inhibitor in Mice

Kim

Papagiannis

Capaldi

et al. 2020

Nucleic Acid Therapeutics

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Safety assessment of drug impurities is a routine part of the drug development process. For oligonucleotide-based drugs, impurities can arise from impurities in starting materials, as by-products of the manufacturing process or from degradation, and are generally structurally similar to the parent oligonucleotide. To study the potential impact of impurities, a representative batch of a 2¢-O-methoxyethyl (MOE) antisense oligonucleotide (ASO) was compared to batches of drug that were enriched with nine of the common impurities encountered with the chemical class. Mice were treated for 3 months with weekly subcutaneous injection of 10 or 30 mg/kg. The impurity content of the parent batch was 0.25%-2.5% of total drug substance. The enriched impurity mixtures contained from 3% to 10% of the various impurities. The expected common class effects were observed at the 30 mg/kg/week dose level in hematology, serum chemistry, and histopathology. However, there were no differences between the representative batch of material and those enriched with impurities. Based on these data, common oligonucleotide impurity studies do not appear to contribute to the overall toxicology profile.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Impurity Qualification Toxicology Study for a 2′-O-Methoxyethyl-Modified Antisense Inhibitor in Mice

Kim

Papagiannis

Capaldi

et al. 2020

Nucleic Acid Therapeutics

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Highway to Success—Developing Advanced Polymer Therapeutics

England

Akhtar

2021

Advanced Therapeutics

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Polymer therapeutics are advancing as an important class of drugs. Polymers have already demonstrated their value in extending the half‐life of proteins. They show great potential as delivery systems for improving the therapeutic index of drugs, via biophysical targeting and more recently with more precision targeting. They are also important for intracellular delivery of nucleic acid based drugs. The same frameworks that have been successfully applied to improve the small molecule drug development can be adopted. This approach together with improved pathophysiological disease knowledge and critical developability considerations, imperative given the size and complexity of polymer therapeutics, provides a structured framework that should improve their clinical translation and exploit their functionality and potential. Progress in understanding the right target, gaining the right tissue and cell exposure, ensuring the right safety, selecting the right patient population is discussed. The right commercial considerations are outlined and the need for a multi‐disciplinary approach is emphasized. Crucial developability factors together with scientific and technical advancements to enable pharmaceutical development of a quality robust product are addressed. It is argued that by applying this structured approach to their design and development, polymer therapeutics will continue to grow and develop as important next generation medicines.

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Evaluation of micropillar array columns for chromatographic separation of phosphorothioated oligonucleotides and their diastereomers

Dillen

Deschrijver

Mol³

et al. 2021

Analytical Science Advances

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Chromatographic analysis of therapeutic oligonucleotides is challenging due to the presence of closely related impurities, degradants or metabolites and due to the presence of phosphorothioate bonds, which introduce chiral centers. In the present study, ion pair reversed phase chromatography of oligonucleotides on micropillar array columns was investigated. Two commonly used mobile phase conditions were included. With 16.3 mM triethylamine and 400 mM hexafluoroisopropanol, the separation of 16‐mer oligonucleotides differing in the number and positions of phosphorothioate linkages as well as some n‐1 and n‐2 truncations demonstrated complete suppression of diastereoselectivity. Although the positional phosphorothioate isomers evaluated could not be resolved, an increase in phosphorothioate bonds resulted in more retention. A therapeutic 19‐mer RNA sequence with 2′‐fluor and 2′‐O‐methyl modifications showed partial separation of some very close impurities. When using 15 mM triethyl ammonium acetate in the mobile phase, diastereomer selectivity was clearly observed for all analytes. The best result was obtained for the 19‐mer RNA therapeutic mimic with four phosphorothioate bonds, since all 16 theoretical diastereomers were clearly observed under the conditions tested. A limited benchmark exercise demonstrated the improved capability of the new micropillar array columns. Therefore, these columns can be positioned as a valuable alternative when challenging oligonucleotide separations are expected.

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Impurities in Oligonucleotide Drug Substances and Drug Products

Cited by 88 publications

References 44 publications

Impurity Qualification Toxicology Study for a 2′-O-Methoxyethyl-Modified Antisense Inhibitor in Mice

Impurity Qualification Toxicology Study for a 2′-O-Methoxyethyl-Modified Antisense Inhibitor in Mice

Highway to Success—Developing Advanced Polymer Therapeutics

Evaluation of micropillar array columns for chromatographic separation of phosphorothioated oligonucleotides and their diastereomers

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