Klotho Neurosciences begins manufacturing KLTO-202 gene therapy to advance clinical trials in ALS

How is Klotho Neurosciences preparing its KLTO-202 gene therapy for investigational ALS trials in 2025?

Klotho Neurosciences, Inc. (NASDAQ: KLTO) announced on June 30, 2025, that it has initiated manufacturing and process development for KLTO-202, its novel gene therapy designed to treat amyotrophic lateral sclerosis (ALS). The New York-based biogenetics innovator is moving toward an Investigational New Drug (IND) application as it prepares for Phase I/II trials slated for the third quarter of 2026. The decision follows two years of extensive preclinical validation involving transgene expression of a neuroprotective RNA splice variant derived from the alpha-Klotho gene.

KLTO-202 uses a proprietary delivery vector designed to introduce a shortened protein isoform known as secreted alpha-Klotho (s-KL) directly into spinal cord neurons. The candidate gene therapy aims to mitigate the oxidative stress and neuroinflammation commonly seen in motor neuron degeneration, the hallmark of ALS. Institutional investors have reacted positively to the move, with analysts pointing to the program’s scientific differentiation and efficient capital deployment strategy that avoids large staffing increases during early-stage trials.

The advancement of KLTO-202 into manufacturing represents a pivotal transition from laboratory findings to clinical application, a milestone closely watched by industry observers given the urgent need for disease-modifying ALS therapies.

What makes the alpha-Klotho splice variant a promising therapeutic target for ALS and neurodegeneration?

The investigational gene therapy KLTO-202 is based on a unique RNA splice variant of the human alpha-Klotho gene, which Klotho Neurosciences licensed from the Autonomous University of Barcelona. The alpha-Klotho gene is known for encoding two major protein isoforms: membrane-bound Klotho (m-KL), found primarily in kidney cells, and secreted Klotho (s-KL), a smaller isoform found in the brain and spinal cord.

The s-KL protein has drawn significant attention in the field of neurodegeneration for its role in modulating oxidative stress, regulating neuroinflammation, and maintaining cellular homeostasis. Overexpression of this variant in animal models has shown neuroprotective effects, especially in conditions characterized by accelerated neuronal loss such as ALS and Alzheimer’s disease. These effects have been documented in peer-reviewed studies involving murine and non-human primate models over the past two years.

By leveraging a gene therapy platform that delivers the s-KL sequence to spinal cord neurons, Klotho Neurosciences hopes to increase endogenous levels of the protein in tissues most impacted by ALS pathology. Preclinical results suggest that elevated s-KL expression may stabilize neuronal function and delay disease progression.

How does the KLTO-202 gene therapy work, and what are the manufacturing goals for 2025?

Klotho Neurosciences is focused on scaling the production of KLTO-202 with the goal of initiating first-in-human studies in late 2026. The product consists of a recombinant adeno-associated virus (AAV) vector encoding the s-KL transgene, optimized to target motor neurons in the spinal cord—cells that deteriorate rapidly in ALS patients.

CEO Dr. Joseph Sinkule emphasized the importance of efficiency in this phase, noting that the gene delivery system has been refined for more effective transduction of target cells. This includes producing higher titers of the AAV vector and improving transgene integration, which is essential for durable protein expression.

The manufacturing roadmap spans approximately eight months and will be conducted in partnership with contract research organizations (CROs), minimizing internal hiring and operating costs. Following this, Klotho Neurosciences will proceed with IND-enabling safety studies, FDA consultation meetings, and clinical site activation.

This lean operating model is designed to reduce the financial burden of scaling early trials, a point that analysts say could make Klotho Neurosciences an attractive partner or acquisition target for larger biotech firms pursuing central nervous system (CNS) pipelines.

What has institutional sentiment indicated regarding KLTO-202’s potential in the competitive ALS treatment landscape?

Investor sentiment toward Klotho Neurosciences has strengthened as the company continues to differentiate KLTO-202 in a field with limited therapeutic breakthroughs. ALS remains a devastating neurodegenerative disease, with most patients facing death within 2–3 years of diagnosis. While FDA-approved treatments such as riluzole and edaravone offer modest benefits, they do not halt or significantly reverse disease progression.

The entry of gene therapies into the ALS pipeline has renewed interest among institutional investors seeking scalable approaches to neurodegenerative disorders. Analysts suggest that KLTO-202’s specificity for spinal cord neurons and its mechanism of amplifying an endogenous protective protein give it a distinct scientific rationale.

Furthermore, Klotho Neurosciences’ capital-light approach to manufacturing and trials—leveraging external partners rather than internal expansion—has drawn attention for its risk mitigation potential. With fundraising efforts reportedly successful, the company is now positioned to meet critical milestones without diluting shareholder equity or compromising operational speed.

What is the historical development timeline of KLTO-202 leading up to 2025 manufacturing?

Klotho Neurosciences’ exploration of the alpha-Klotho gene began shortly after its 2022 licensing deal with the Autonomous University of Barcelona. That agreement gave the American biogenetics company exclusive access to patent families, research data, and platform technology around the s-KL splice variant. Over the next two years, the firm funded animal studies across three disease models: accelerated aging, Alzheimer’s disease, and ALS.

These studies demonstrated that transgene overexpression of s-KL improved neurological markers, reduced oxidative stress, and stabilized neuronal survival in both murine and primate systems. Peer-reviewed publications confirmed that AAV-mediated gene therapy with s-KL produced consistent and durable protein expression in CNS tissues.

By early 2025, Klotho Neurosciences had finalized its vector optimization strategy and began process development for scale-up manufacturing. The transition to clinical preparation was enabled by recent financing rounds and strategic partnerships with CROs experienced in gene therapy development.

What are the regulatory and clinical milestones expected for KLTO-202 in the coming year?

Following the current eight-month manufacturing timeline, Klotho Neurosciences plans to initiate FDA discussions and complete all mandatory preclinical safety studies. These efforts are projected to span four to six months, during which the company will also finalize its Investigational New Drug (IND) filing.

Clinical site identification and staff training will run in parallel, with the goal of enrolling ALS patients in a Phase I/II single-dose escalation study by Q3 2026. This first-in-human trial is expected to evaluate safety, tolerability, and preliminary efficacy in a small cohort of individuals with early-stage ALS.

The trial design will likely follow a standard gene therapy protocol, involving low-to-high dose escalation and extended post-treatment monitoring. While KLTO-202 remains in the preclinical stage, institutional expectations are high given the strong animal model data and streamlined development approach.

How does KLTO-202 compare to other emerging ALS therapies in the 2025 biotech pipeline?

KLTO-202 enters a gene therapy pipeline that is gradually gaining momentum, particularly after the FDA’s 2023 approval of Biogen’s QALSODY for SOD1-mutated ALS. While QALSODY targets a specific genetic subset, KLTO-202 is designed for broader application by enhancing s-KL protein levels across both familial and sporadic ALS cases.

Other companies in the ALS gene therapy space include Amylyx Pharmaceuticals and Wave Life Sciences, both of which are pursuing RNA- or DNA-based approaches. However, KLTO-202’s mechanism is unique in that it amplifies a naturally occurring neuroprotective protein rather than silencing or editing pathogenic genes.

Given that s-KL expression is reduced in aging and neurodegenerative tissues, the therapy may have potential applications beyond ALS, including in Alzheimer’s and Parkinson’s disease. Klotho Neurosciences’ broader pipeline includes genomics-based diagnostics and other gene-modified cell therapies targeting age-related neurological decline.

What is the long-term outlook for Klotho Neurosciences’ gene therapy strategy beyond ALS?

Beyond KLTO-202, Klotho Neurosciences has built a platform around alpha-Klotho biology, aiming to develop gene and cell therapies that address other forms of neurodegeneration. The company’s pipeline includes exploratory work in Alzheimer’s disease, where s-KL expression has been shown to protect against tau aggregation and amyloid toxicity in preclinical models.

With a focus on aging-related disorders, the American biogenetics developer seeks to expand into Parkinson’s disease and possibly rare leukodystrophies, leveraging the same gene delivery framework. Institutional analysts suggest that if KLTO-202 achieves safety validation and preliminary efficacy in ALS patients, it could pave the way for additional indications using the same transgene backbone.

Moreover, the scalability of the s-KL platform—combined with Klotho Neurosciences’ efficient use of external manufacturing and CROs—may allow the company to enter strategic partnerships or pursue fast-track regulatory pathways, especially under orphan disease designation programs.