Industrial-Scale Manufacturing of a Possible Oligonucleotide Cargo CPP-Based Drug

Tedebark, Ulf; Scozzari, Anthony N.; Werbitzky, Oleg; Capaldi, Daniel C.; Holmberg, Lars

doi:10.1007/978-1-60761-919-2_36

Cited by 8 publications

(10 citation statements)

References 18 publications

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“…A four time excess of amidite was needed to obtain almost 100% coupling efficiency. In real oligonucleotide synthesis, amidite excesses between 1.5 and 2.0 are routinely used in flow through synthesizers . In batch synthesizers the excess is usually between 4.0 and 10.0 2 .…”

Section: Resultsmentioning

confidence: 99%

“…In real oligonucleotide synthesis, amidite excesses between 1.5 and 2.0 are routinely used in flow through synthesizers. 8 In batch synthesizers the excess is usually between 4.0 and 10.0 2 . The results of the simulation are therefore in a very realistic range.…”

Section: Scale-upmentioning

confidence: 99%

“…In the field of molecular biology enormous numbers of oligonucleotides are synthesized parallel in smallest amounts in situ on microarrays for gene analysis . On the other hand kilogram quantities are needed for therapeutic applications and nontherapeutic applications such as anticounterfeiting markers . Both approaches need specially engineered equipment, processes, and also raw materials.…”

Section: Introductionmentioning

confidence: 99%

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Optimization and scale‐up of oligonucleotide synthesis in packed bed reactors using computational fluid dynamics modeling

Wolfrum¹,

Josten

Götz

2014

Biotechnology Progress

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A computational fluid dynamics (CFD) model for the analysis of oligonucleotide synthesis in packed bed reactors was developed and used to optimize the scale up of the process. The model includes reaction kinetics data obtained under well defined conditions comparable to the situation in the packed bed. The model was validated in terms of flow conditions and reaction kinetics by comparison with experimental data. Experimental validation and the following model parameter studies by simulation were performed on the basis of a column with 0.3 g oligonucleotide capacity. The scale-up studies based on CFD modelling were calculated on a 440 g scale (oligonucleotide capacity).

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Section: Resultsmentioning

confidence: 99%

Section: Scale-upmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Optimization and scale‐up of oligonucleotide synthesis in packed bed reactors using computational fluid dynamics modeling

Wolfrum¹,

Josten

Götz

2014

Biotechnology Progress

View full text Add to dashboard Cite

show abstract

“…This is of the utmost importance in outbreak response and management efforts, as the speed at which a therapeutic can be Picornaviridae Positive-sense singlestranded RNA genome 3D protein and 3 0 untranslated region developed and produced in large quantities for treatment is often a major bottleneck to management strategies. Industry has also recognised this need and have now made the fast translation of large quantities of therapeutic product feasible [55,56]. For instance, in the case of the recent Ebola outbreak, a newly designed siRNA targeting the emerged Ebola virus Makona strain was produced and incorporated into a lipid NP system within 8 weeks [57], and was deployed for compassionate use during the outbreak.…”

Section: Sirna As An Antiviral Therapeuticmentioning

confidence: 99%

Polymers in the Delivery of siRNA for the Treatment of Virus Infections

2017

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Viral diseases remain a major cause of death worldwide. Despite advances in vaccine and antiviral drug technology, each year over three million people die from a range of viral infections. Predominant viruses include human immunodeficiency virus, hepatitis viruses, and gastrointestinal and respiratory viruses. Now more than ever, robust, easily mobilised and cost-effective antiviral strategies are needed to combat both known and emerging disease threats. RNA interference and small interfering (si)RNAs were initially hailed as a "magic bullet", due to their ability to inhibit the synthesis of any protein via the degradation of its complementary messenger RNA sequence. Of particular interest was the potential for attenuating viral mRNAs contributing to the pathogenesis of disease that were not able to be targeted by vaccines or antiviral drugs. However, it was soon discovered that delivery of active siRNA molecules to the infection site in vivo was considerably more difficult than anticipated, due to a number of physiological barriers in the body. This spurred a new wave of investigation into nucleic acid delivery vehicles which could facilitate safe, targeted and effective administration of the siRNA as therapy. Amongst these, cationic polymer delivery vehicles have emerged as a promising candidate as they are low-cost and easy to produce at an industrial scale, and bind to the siRNA by non-specific electrostatic interactions. These nanoparticles (NPs) can be functionally designed to target the infection site, improve uptake in infected cells, release the siRNA inside the endosome and facilitate delivery into the cell cytoplasm. They may also have the added benefit of acting as adjuvants. This chapter provides a background around problems associated with the translation of siRNA as antiviral treatments, reviews the progress made in nucleic acid therapeutics and discusses current methods and progress in overcoming these challenges. It also addresses the importance of combining physicochemical characterisation of the NPs with in vitro and in vivo data.

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“…Thirdly, the SPOS process is highly mass intensive at large scale 6 . For instance, to produce 1kg of 20-mer phosphorothioate oligos, approximately 4,000 kg of solvents, reagents, and water are needed 16 .…”

Section: Introductionmentioning

confidence: 99%

Organic Solvent Nanofiltration (OSN): A New Technology Platform for Liquid-Phase Oligonucleotide Synthesis (LPOS)

Kim

Gaffney

Valtcheva

et al. 2016

Org. Process Res. Dev.

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Organic Solvent Nanofiltration (OSN) technology is a membrane process for molecular separation in harsh organic media. However, despite having well-documented potential applications, development hurdles have hindered the widespread uptake of OSN technology. One of the most promising areas of application is as an iterative synthesis platform, for instance for oligonucleotides or peptides, where a thorough purification step is required after each synthesis cycle, preferably in the same working solvent. In this work, we report a process development study for liquid-phase oligonucleotide synthesis (LPOS) using OSN technology. Oligonucleotide (oligo) based drugs are being advanced as a new generation of therapeutics functioning at the protein expression level. Currently, over one hundred oligo based drugs are undergoing clinical trials, suggesting that it will soon be necessary to produce oligos at a scale of metric tons per year. However, there are as yet no synthesis platforms that can manufacture oligos at >10 kg batch-scale. With the process developed here, we have successfully carried out 8 iterative cycles of chain extension and synthesized 5-mer and 9-mer 2'-O-methyl oligoribonucleotide phosphorothioates, all in liquid phase media. This paper discusses the key challenges, both anticipated and unexpected, faced during development of this process, and suggests solutions to reduce the development period. An economic analysis has been carried out, highlighting the potential competitiveness of the LPOS-OSN process, and the necessity for a solvent recovery unit.

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Industrial-Scale Manufacturing of a Possible Oligonucleotide Cargo CPP-Based Drug

Cited by 8 publications

References 18 publications

Optimization and scale‐up of oligonucleotide synthesis in packed bed reactors using computational fluid dynamics modeling

Optimization and scale‐up of oligonucleotide synthesis in packed bed reactors using computational fluid dynamics modeling

Polymers in the Delivery of siRNA for the Treatment of Virus Infections

Organic Solvent Nanofiltration (OSN): A New Technology Platform for Liquid-Phase Oligonucleotide Synthesis (LPOS)

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