New immunomodulatory drugs have the potential to cause serious immune-related adverse events, including cytokine release syndrome (CRS). CRS is a systemic inflammatory response characterized by the release of
pro-inflammatory cytokines from immune cells, which results in fever, fatigue, and possibly multiple organ failure.

In Issue 44 of The Altascientist, we review strategies, including those implemented at Altasciences, for developing, qualifying, and implementing cytokine release assays (CRAs) to assess the safety of test articles in vitro to complement in vivo preclinical safety assessments, in addition to: 

• the importance of nonclinical cytokine release evaluation
• customized approaches to CRA use in drug development
• top five aspects to consider before running a CRA
• Altasciences’ case results
• how Altasciences can help 

Drugs that are likely to require CRAs  as part of derisking their development include, but are not limited to:

• immune checkpoint inhibitors
• gene therapies
• oncolytic viral therapies
• cytokine-based therapies
• nanoparticle-based therapies
• therapeutics targeting autoimmune diseases
• vaccines (traditional and mRNA)
• antibody-drug conjugates (ADCs)
• chimeric anti gen receptor cells (e.g., CAR T cells)

WHAT IS THE BENEFIT OF NONCLINICAL CYTOKINE RELEASE EVALUATION? 

Nonclinical safety assessment serves as a cornerstone in drug development, providing essential data to identify early signs of potential toxicity and inform subsequent clinical trial design. One important aspect of toxicity studies is determining whether the drug produces undesired immune effects. The focus is to ensure that the drug does not have any potential to induce an excessive immune response, particularly cytokine storm, which can lead to severe adverse effects and impede clinical development.

During nonclinical safety assessment, we investigate whether there is an increase or decrease in cytokine production in the serum of dosed animals compared to pre-dose and vehicle controls over time. If there are indications in vivo alluding to a trend, such as a dose-dependent increase of specific cytokines in several or all treated animals, a CRA may be conducted to confirm these results in vitro.

Alternatively, if a drug’s mechanism of action or the patient population being targeted poses a higher risk for undesired immune effects, the CRA may be included in the development pathway prior to starting first-in-human studies, even in the absence of any immunotoxicity trends during nonclinical safety testing.

Customized Approaches to Cytokine Release Assay Use in Drug Development

In a CRA, the test article or antibody therapeutic is co-cultured with human immune cells such as peripheral blood mononuclear cells (PBMCs), whole blood, or other appropriate cells . The amount of inflammatory cytokines produced is measured and provides critical data for hazard identification.

Although there is no published regulatory guidance specific to CRA development, validation, or use, the industry has developed several approaches in performing the CRA assay based on the drug type and its mechanism of action, including:

• PBMC  Assays or Whole Blood Assays
• Solid Phase (SP) or Aqueous Phase (AQ) Formats
• Choosing the Right Cytokines to Test
• Appropriate Timepoints for Cytokine Measurement
• Choosing the Appropriate Concentration of Drug Product
• Appropriate Controls for Cytokine Release Assays

Read Issue 44 to learn more about these approaches to performing CRAs.  

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Current and future development of glucagon-like peptide-1 receptor agonists (GLP-1 RAs), or simply GLP-1s, involves complexities not only in formulation but in all areas of early-phase drug development. Developing and refining preclinical models for early efficacy signals, developing and validating the bioanalytical assays necessary for quantitation, and designing the clinical studies that deliver the most robust data in this innovative therapeutic area are all key elements of a drug development program.

In Issue 42 of The Altascientist, we review the requirements for successful GLP-1 drug development, including:

• preclinical approaches 
• early-phase clinical study design 
• bioanalytical techniques 
• manufacturing 

Two case studies are also included. 

The Golden Era of GLP-1 Drugs

GLP-1 RAs are entero-pancreatic hormone-based treatments, first approved in 2005 for type 2 diabetes (T2D), and now commonly prescribed for weight loss and reduction of atherosclerotic cardiovascular (CV) risk in patients with T2D.

According to a MarketsandMarkets report from July 2024, the global GLP-1 market is expected to reach $471 billion worldwide by 2032. More than one billion people worldwide are living with obesity, which increases their risk of major health complications, including T2D, high blood pressure, heart disease, stroke, metabolic syndrome, fatty liver diseases, some cancers, kidney disease, breathing problems, and sleep apnea. Most of these diseases are interrelated, with glycemic control and weight loss being a major factor in improving the overall health of sufferers.

“As we learn more about GLP-1 RAs, we realize that they target many organ systems, including the pancreas, the stomach, brain, heart, kidneys, immune system (due to reduced inflammation), and skeletal muscle. They help control the metabolism of white and brown adipose tissue and show positive effect in fatty liver disease (nonalcoholic steatohepatitis, or NASH),” says Dr. Gaetano Morelli, Chief Medical Officer at Altasciences.

What We Know So Far About GLP-1 RAs

GLP-1 RA head-to-head clinical studies have demonstrated that all GLP-1 RAs are effective at reducing A1C (the average amount of glucose in a person’s blood over the past three months). The initial pivotal studies that led to the first wave of GLP-1 RAs demonstrated that they work in several ways, including stimulating insulin release, slowing digestion, reducing appetite, and inhibiting glucagon release. These actions have made GLP-1 RAs excellent candidates for obesity treatment, where much of the new development is now focused. As with most medical breakthroughs, the development and implementation of GLP-1 RA drugs is not straightforward. 

How Altasciences Can Help With Your GLP-1 Drug Development