How the Program Took Shape
The work began several years ago when Dr. Sinkule founded Anew Medical, a holding company for in-licensed medical development assets. While reviewing preclinical work in ALS and Alzheimer’s models and monitoring the literature surrounding the gene of interest, the scientific direction became clear. He and his team then realigned their efforts around developing a gene-based therapeutic approach. They renamed the company, Klotho Neurosciences, Inc.
As he explained, “Our gene therapy approach is novel, it’s patented now and the preclinical data across several different neurodegenerative diseases is compelling.”
The program centered on a protein he described as “neuroprotective”, with activity relevant to several indications. Levels of the klotho protein naturally decline with age as the gene becomes increasingly silenced. This gene was called “klotho.” Restoring those levels through a gene-based construct — rather than repeated external infusions — became the foundation of the strategy.
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Why the Klotho Gene?
Interest in the gene grew as Dr. Sinkule reviewed preclinical work showing protective effects in models of neurodegenerative disease, alongside a large body of scientific literature describing its role in aging biology. Levels of the protein naturally fall over time as the gene and promoter become increasingly silenced due to hypermethylation, which creates a therapeutic opportunity.
— Dr. Joseph Sinkule, Founder and CEO, Director and Chairman of the Board, Klotho Neurosciences
Why Neurology Trials Remain Challenging
Before entering the neurodegenerative space, much of Dr. Sinkule’s early work centered on oncology. That background shaped how he viewed differences between the two fields.
Oncology trials traditionally rely on established, objective endpoints. Neurology trials, by contrast, often depend on observational tools or symptom-based assessments.
“Many of the clinical trials in neurologic diseases are, in my opinion, fairly subjective based on diary entries and someone’s interpretation,” he said.
“It was very challenging to quantify the endpoints of neurodegenerative diseases other than stopping the progression or somehow improving the lifespan,” he added.
ALS differs slightly because of its rapidly fatal course — patients often die within two to three years of diagnosis — which makes survival a more concrete endpoint.
Dr. Sinkule noted that biomarkers such as neurofilament light chains and certain PET imaging measures are now being used in trials and accepted by regulators as early indicators of efficacy. However, he cautioned that some therapies cleared on biomarker data later failed to show the same benefit in larger post-approval studies.
What Makes the Next Generation of Studies More Promising
When discussing why he feels optimistic about the next wave of ALS studies, Dr. Sinkule pointed to preclinical observations. In animal models carrying a human SOD1 mutation, a mutation that is rare in ALS but clinically relevant, his team saw signs of improved muscle function and other indicators of activity following a single dose of the gene-based construct.
He said, “We’ve been able to improve the survival endpoint by 400% in animal models treated with a single dose of the gene therapy… and we’re assuming that human klotho in humans with the SOD1 mutation again results in this increase in survival.”
He added that maintaining muscle strength and contractility are key early indicators to watch before longer-term endpoints, such as survival, are reached. The design of their upcoming clinical studies, he noted, reflects this stepwise approach. “I think we have a very interesting clinical trial approach,” he said.
How Biomarkers Are Being Used Today
Biomarkers are gaining traction in neurodegenerative research, though their role is still evolving. Dr. Sinkule noted that some programs advanced based on biomarker signals, only for larger, required Phase IV studies to show weaker-than-expected performance.
As he explained, “some of the biomarkers led to an approval and then the FDA required a larger Phase IV follow-up study… and the products failed to perform as they did with the biomarkers.”
Because of this uncertainty, his team plans to use an adaptive trial design. This approach allows them to incorporate biomarkers early but continue enrollment to reach a patient sample size large enough to evaluate endpoints such as non-inferiority or improvement over current care. Dr. Sinkule described this as a way to align early biomarker readouts with the longer-term clinical outcomes needed for ALS studies.
— Dr. Joseph Sinkule, Founder and CEO, Director and Chairman of the Board, Klotho Neurosciences
Delivery Barriers: Reaching the Brain and Spinal Cord
One of the largest obstacles in neurology is delivering therapies to the brain and spinal cord. The blood-brain barrier (BBB) acts as a protective system, limiting access to many biologics and gene-based constructs.
Researchers have explored a range of strategies to overcome this barrier, including ligand-receptor pathways such as transferrin or insulin to help move molecules from the bloodstream into the central nervous system.
Dr. Sinkule’s own program uses an adeno-associated virus (AAV) as the delivery platform. He explains that this vector can infect neurons and support local production of the Klotho protein within the nervous system. He also noted that traditional AAV serotypes can accumulate in the liver, creating safety challenges in some gene therapy trials.
To address this, his team began working with AAVnerGene, which developed a variant capable of binding and crossing the BBB. This variant is intended for their Alzheimer’s and Parkinson’s constructs, while a different, muscle-tropic version is being used for ALS. He noted that Klotho levels are normal early in life but become nearly undetectable by middle age as the gene is gradually silenced through hypermethylation. The company’s therapeutic strategy, he added, is not to cut or replace the gene, but to provide a genetic construct of the klotho gene that allows the body to make the protein again on its own.
Manufacturing and Scalability: A Complex Process
Manufacturing gene-based therapies remains resource-intensive. The process spans construct design, process development, production and extensive testing.
“Gene therapy to make a clinical batch… is around $5 million to $6.5 million… and that can take nine months to 12 months and is very labor intensive,” he explained.
To address these challenges, newer production platforms are emerging. Dr. Sinkule described a cell-engineering approach that embeds all components required to make the capsid directly into a 293-derived line, allowing developers to introduce only the gene of interest rather than multiple plasmids. This reduces the uncertainties associated with multi-plasmid transfection and the number of variables that can fail during production.
As he described it, “Instead of using three plasmids to transfect into a cell, you basically have a cell that is now just introducing a single plasmid containing your gene of interest.”
Dr. Sinkule also highlighted the importance of vector variants that concentrate in targeted organs rather than accumulating in the liver. These advances may further streamline the path to clinical material by enabling more precise, tissue-focused delivery strategies
Dr. Sinkule closed with a quick but conclusive note on gene therapy: “It starts with making DNA and all these other steps beyond just making the DNA plasmid that effectively expresses the protein in the body over long durations of time.” Dr. Sinkule claims “gene therapy is the future of medicine” and we will continue to be focused on DNA-based products that are safe and effective.”
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