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Feliza Mirasol

®, Matthew Lauber, senior director—Cell and Gene Therapy, Consumables & Lab Automation, Waters Corporation, had this to say when asked his thoughts on whether oligonucleotides (oligos) may be the next wave of biotherapeutics on the market in the near future: “The market for synthetic oligonucleotides is happening right now,” as he pointed out that oligos historically got their start in the late 1990s.

“What's interesting,” he notes, “is that the delivery mechanisms have been improving as of late.” Lauber points to some of the earliest oligo drugs using lipid nanoparticles as a delivery mechanism, sharing that “the basis of that technology is what [also] gave the formulation for the [messenger RNA] vaccines.”

However, he adds, today, the synthetic oligo market has moved on and is not using these nanoparticles anymore, which he characterizes as an amazing technology in and of themselves. Rather, he explains, whatever is needed for drug delivery is being put directly onto the synthetic oligo, so that, as in the case of a lipid modification, a direct conjugate is added directly onto the oligo molecule that helps the drug be internalized.

“All of this [new technology] has given [the] basis for this resurgence, and there are hundreds of new synthetic oligos in the market right now. It's an interesting subsegment to the overall wave coming up nucleic acid-based medicine,” Lauber states.

AAPS PharmSci 360 ran Oct. 22–25 in Orlando, Fl.

 industry’s focus on oligonucleotide therapeutics

Articles

Matthew Lauber, senior director—Cell and Gene Therapy, Consumables & Lab Automation, at Waters Corporation, considers that status of oligonucleotides on the market.

Oligonucleotides: The Next Therapeutic Wave on the Market? (AAPS PharmSci 360)

“You might be familiar with Waters. Waters is a specialty instrument company focused on making analytical measurements, and with this analytical science, our team has been studying what would be needed to help improve the quantitation reliability and robustness quantitation of synthetic oligonucleotides from biofluids,” says Matthew Lauber, senior director—Cell and Gene Therapy, Consumables & Lab Automation, Waters Corporation in an exclusive interview with 

Traditionally, Lauber says, improving such analytical techniques has been done by liquid–liquid extraction, which is often involved in the use of toxic chemicals such as chloroform and phenol, which are iteratively used for liquid–liquid extraction. These techniques involve laborious procedures, but, today, people in labs are wanting to have a safer environment and to move toward greener chemistries, he emphasizes.

“What we've devised here [with Waters’ new bioanalysis kit] is an approach that takes advantage of enzyme technology, or recombinant proteinase K, that will act upon the protein composition of any biofluid or tissue sample. So, this rapidly degrades protein composition, and this helps us work against what was designed in the oligonucleotide for its pharmacology effects,” Lauber states. He further explains that oligonucleotides with lipid modifications and backbone modifications are designed to bind to proteins, but that action is essentially an interference on the quantitation assay.

“Secondly, after a rapid digestion of all the proteome, we can take the sample and load to a mixed-mode SPE (solid phase extraction) procedure, and this is a specially tuned type of chemistry, where we bind the washing of the glue and the resulting sample; we produce something that gives high recovery, high yield of the oligonucleotide drugs and their metabolites,” Lauber continues. “This facilitates, directly, a direct injection into an LC–MS [liquid chromatography–mass spectrometry] experiment where triple quad mass spectrometry can be used for very sensitive analysis of the sample. This produces a PK/PD [pharmacokinetic/pharmacodynamic] curve. You can also give information on, of course, half-lives and the metabolites of disease,” he concludes.

nd the industry’s focus on oligonucleotide therapeutics

Matthew Lauber, senior director—Cell and Gene Therapy, Consumables & Lab Automation, at Waters Corporation highlights the company’s motivation behind their new bioanalysis kit for oligonucleotides.

The Need for New Bioanalysis Technology (AAPS PharmSci 360)

® at AAPS PharmSci 360, Matthew Lauber, senior director—Cell and Gene Therapy, Consumables & Lab Automation, Waters Corporation, discusses the analytical and downstream purification challenges of working with oligonucleotides (oligos). He notes that the pharmaceutical landscape is changing—diversifying into more types of molecules.

Of oligos, he says, “Synthetic oligonucleotide therapies represent a type of molecule that's a little bit in between both worlds, being that they’re synthetically produced, yet they are really large molecules in a way —they're 15- to 30-mer residue long oligonucleotides. But being that they're chemically synthesized, what is occurring right now in the landscape—in the pipeline—is they're being modified extensively with phosphorothioate backbones, with methyl group modifications, and, on top of this, they're being conjugated with new saccharide and lipid moieties so that they can be very carefully controlled for their absorption.”

Meanwhile, commenting on some key challenges in the downstream purification of oligo molecules, Lauber explains that “because they are so exquisitely modified, these chemically synthesized oligonucleotides also need to be carefully purified, and this is usually done through anion exchange or first-phase preparative chromatography.”

“With this,” he continues, “the impurities have to be analyzed to at least 0.1% relative abundances and thoroughly understood. Many companies are working very hard on characterizing those impurities.”

Lauber goes on to explain that, even with a purified drug substance, however, there is another type of analysis—quantitative bioanalysis—that is important. This quantitative bioanalysis enables researchers to “see what happens with the drug once it's dosed to an animal model, or first studied in 

 cell analysis, or later on in clinical study as the first-in-humans study.”

View Lauber’s video interviews about the 

industry’s focus on oligonucleotide therapeutics

Matthew Lauber, senior director—Cell and Gene Therapy, Consumables & Lab Automation, at Waters Corporation discusses the analytical and downstream purification challenges of oligonucleotides.

Discussing Oligonucleotide Challenges in Analytics and Purification (AAPS PharmSci 360)

® at AAPS PharmSci 360, Dujuan Lu, PhD, global leader—E&L, SGS Health Sciences, reviewed the difference between establishing extractables and leachables (E&L) testing for combination products and conventional single/simplified drug products.

“If you only have just a drug product,” she explains, “you only need to consider the extractables and leachables portions for a drug product or regulation. You know the pharmaceutical packaging perspective or maybe the manufacturing process material as well.”

“However,” she continues, “if you are dealing with drug­device combination products, you need to consider both aspects, you need to work on the drug perspective and also the device perspective. And this [will] really depend on the primary mode of action, [such as] whether you need to put more focus on the drug side or more focus on device side.”

A combination drug product is more complicated and definitely requires consideration for both aspects, she states. “In my talk, we're talking about different regulations for different types of study design in terms of the drug versus device. So, all aspects need to be consideredduring the study design for combination products.”

Lu was featured as a speaker at AAPS PharmSci 360, where she gave a talk, “

Regulatory Expectations in Extractables and Leachables Testing of Combination Products

.” In her talk, she discussed chemical characterization, per ISO 10993-18, which has become important in biocompatibility testing for medical devices. One of the key points in her talk was on the differences and similarities of E&L study designs between medical devices (based on ISO standards) and drug product ( based on USP standards). She also discussed the need for understanding the recent regulatory expectations on E&L testing of medical devices and combination products.

Dujuan Lu, PhD, global leader—E&L, SGS Health Sciences, talks about the differences in designing E&L testing for combination products vs. conventional pharmaceuticals.

Distinguishing E&L Testing for Combination Products (AAPS PharmSci 360)

® at AAPS PharmSci 360, Dujuan Lu, PhD, global leader—E&L, SGS Health Sciences, summarizes some of the points in her presentation at the conference, mainly about the differences and similarities between extractables and leachables (E&L) study design between ISO and USP standards.

For instance, she points out that for E&L studies being conducted on medical devices, then the standards from ISO 10993-18 for the chemical characterization of medical devices, whereas E&L studies being done on pharmaceuticals will typically follow USP <1663>/<1664> guidance for pharmaceutical packaging, such as the finished packaging/container closure system, and USP <665>/<1665> for the pharmaceutical manufacturing component.

The major difference between the two, she emphasizes, is that there are different extractables that may be tested. For instance, for a medical device, ”you might need to do an exhaust extraction for implants, while, for the pharmaceutical, you don't really need to do that,” Lu states. In addition, a different type of solvent may be used on a medical device, such as some harsher solvents (e.g., hexane, isopropanol, etc.).

However, in terms of analytical testing between ISO and USP, Lu says that they are similar, such as gas chromatography with mass spectrometry (GC–MS).“There is GC–MS for volatiles; there is direct injection GC–MS for semi volatiles, LC–MS [liquid chromatography with mass spectrometry] for the nonvolatiles, and the ICP–MS [inductively coupled plasma mass spectrometry] for elemental impurities,” she says.

 and the distinction between E&L testing for combination products vs. conventional drug products.

Dujuan Lu, PhD, global leader—E&L, SGS Health Sciences, discusses the differences in how E&L study design should be approached based on whether the study  is following ISO or USP standards. 

Discussing E&L Study Design Between ISO and USP (AAPS PharmSci 360) 

® at AAPS PharmSci 360, Dujuan Lu, PhD, global leader—E&L, SGS Health Sciences, gave her views on what key extractables and leachables (E&L) issues pharma manufacturers should keep in mind, from a regulatory standpoint, when formulating a pharmaceutical that requires a medical device for drug delivery.

She points out that, for any drug product that uses a device as a drug delivery system, a pharma manufacturer will need to consider extractable/leachable testing for both aspects. “You need to conduct your extractables and leachables study for your container closure system for the drug product, but, on the other hand, you need to work out your extractables and leachables for your drug delivery device,” she says.

That that end, a pharma manufacturer may need to consider the regulations in place for that device and then design its E&L study that is applicable to that portion. “You need to consider both aspects in that case,” Lu recommends.

Dujuan Lu, PhD, global leader—E&L, SGS Health Sciences, recommends some key issues that pharma manufacturers should consider when formulating products that require a medical device.