Complement pathways are well characterized as a part of the larger immune system. Recent published data suggest larger roles for complement cascades, particularly in neurobiology. Complement pathways are upregulated in transcriptome databases generated by the ALS Therapy Development Institute. These data have informed a series of projects designed to exploit complement pathways as therapeutics targets for ALS.

ALS is a disease that results in the dysfunction of motor units and the eventual death of motor neurons. It is possible that for a variety of reasons motor neurons in ALS are less capable of responding to stress caused by over-activity, oxidative damage, or misfolded proteins, etc. A variety of therapeutic strategies are being tested to reduce stress or bolster the vulnerable cells' natural stress response mechanisms.

For decades, many researchers and clinicians have suspected that the immune system plays a role in the pathophysiology of ALS. Many hypotheses about this role have been proposed over the years. It is possible that specific components of the immune system are protective and compensatory in ALS while dysregulation of other immune system components are deleterious. Striking the right balance with treatments presents significant challenges. The ALS Therapy Development Institute's has created comprehensive transcriptome databases to help to guide the prioritization of immunomodulatory therapeutic strategies that are being tested internally.

Trophins are proteins that have been shown through in vitro or in vivo systems to help neurons survive, differentiate, or grow. Because ALS is a disease where motor neurons die, neurotrophins may be beneficial for therapeutic development. A variety of trophins and trophin delivery strategies are being tested at the ALS Therapy Development Institute for therapeutic benefit.

Unlike the other defined research programs, "Knockdown" does not have a single specific class of biological target pathways. Rather "Knockdown" is the name of a program designed to develop technologies that can best be used to suppress RNA expression of genes that are known or suspected to cause or propogate the ALS disease state. Genes like mutant SOD and TDP-43 may be targets for this technological development.

Recently, data have suggested that motor unit dysfunction in ALS may begin at the axon and at synaptic connections. Various strategies for protecting the myelin sheath, remyelination, and synaptic remodeling are being studied at The ALS Therapy Development Institute. Researchers at the ALS Therapy Development Institute have the imaging expertise to characterize anatomical changes at the neuromuscular junction and determine whether or not a treatment is protecting the site.

Researchers at the ALS Therapy Development Institute come from a variety of backgrounds and often have different and complementary areas of expertise. These different perspectives on the data generated internally, and work that is published by the external research community, leads to many different hypotheses about how to attack ALS. A new hypotheses presented internally is called a New Lead.

New Leads are presented regularly at Research Staff meetings. Staff scientists discuss and formulate hypotheses about each therapeutic strategy, focused on what could lead to treatments for ALS. Scientists are asked to present these ideas and any supporting data. The idea is then debated and potential strengths and pitfalls are addressed. When additional information is needed, the submitting scientists may be asked to gather more information and resubmit. In some case, the New Lead is rejected as a Project because there is a lack of supporting evidence to justify the high expense of initiating experiments.

Learn more about the science team at ALS TDI.

Generally speaking, formulation is the process by which different chemical substances are combined to produce a final, potential medicinal product which can be tested. For our purposes, “Formulation” is used more broadly as an umbrella term to characterize the stage of therapeutic development where the treatment is being manufactured, purified, and prepared for experimentation in cell culture and/or in vivo. There are several different types of medicinal products which can be created in order to deliver and affect the intended pathway; the formulation process for the four bring used at the ALS Therapy Development Institute are described here. Each has both advantages and disadvantages and it is not uncommon for more than one type of product to be created for a target pathway:

Small Molecule Drugs
In the cases of small molecule drugs, Formulation includes acquisition or synthesis of the active drug. In most cases, so the drug can be administered in vitro/in vivo, the small molecule drug combined with solvents so that precise doses can be delivered. At the ALS Therapy Development Institute, small molecule drugs can be delivered in vivo using various routes of administration:

• Orally in drinking water
• Orally in food
• Orally by gavage (syringe administration)
• Subcutaneous injection
• Intraperitoneal injection
• Subcutaneous mini-pump implantation
• Intrathecal infusion for delivery of the test compound directly to the spinal cord
• Intracerebral ventricular infusion for delivery of test compound directly to the cerebral spinal fluid around the brain

Biologics
This class of therapeutics must either be engineered and manufactured internally or acquired from a third party. Unlike small molecule drugs that can be synthesized chemically, biologics generally require biological systems to be generated. Because of this, close attention is directed to ensure the purity, integrity, and biological activity of the therapeutic.

Gene Therapy
Gene Therapies must be designed and engineered internally prior to being manufactured by third party vendors at a scale adequate to the needs of the rigorous in vitro and in vivo experimentation executed at that ALS Therapy Development Institute. Similar to biologics, gene therapies typically rely on biological systems to be generated. If a biological system is used, the resulting gene therapeutics must be further tested and monitored for purity and effectiveness of the “vector”; the biological or synthetic tool used to deliver the genetic material. The ALS Therapy Development Institute has established relationship with several vector producing organizations, amassing the ability to ramp up several different types of gene therapy products simultaneously.

Stem Cell Therapeutics
Stem Cell therapies and cell-based therapies in general present challenges different from each of the other categories because of the undefined nature of these emerging technologies. Purity and cellular viability must be monitored closely in order to produce interpretable results. Additionally, the nutrient rich fluid, or media, in which cells are cultured and delivered, plays a key role in defining how the cells differentiate. Additionally, each component of the nutrient rich fluid could have a positive or negative effect in an experiment. Therefore, the media also needs to be monitored for purity, sterility, and overall content in order to generate interpretable results. While little is known about the most likely therapeutic use for stem cells, researchers have used stem cells as an in vitro research tool to validate therapeutic activity in a specific cell type and as the biological delivery vehicles for gene therapies and other potential therapeutics which require a biological transfer medium.

In vitro validation refers to the process by which any therapeutic agent is tested in cell culture systems to confirm that the therapeutic actively impacts its biological target with a desired degree of specificity. In vitro systems reduce the number of animals used in therapeutic development and can increase the pace of research. These experiments are relatively quicker to execute than in vivo experiments and many different cell lines can be created in the lab and used to test a potential therapeutic. While it is a critical step in the drug development process, in vitro validation is limited in that it typically can only be used to characterize the therapeutic effect on one cell type at a time and often cannot model more complex systems where different cell types interact with each other in diseased or healthy states.

Pharmacokinetics is the study of what the body does to a drug or therapeutic over time. Pharmacokinetic studies characterize the absorption, distribution, metabolism, and elimination of a therapeutic by the body. Pharmacodynamics is the study of the effect a therapeutic has on the body's physiology; in particular in relation to dose and drug concentration. The execution and analysis of pharmacokinetic studies, in tandem with pharmacodynamic studies in vivo, typically defines the dosing regimen that will be used in survival efficacy studies.

Survival Efficacy studies are executed at the ALS Therapy Development Institute the animal model of disease. These in vivo experiments are designed to identify therapeutics that extend survival, delay paralysis, improve neurological symptoms, and allow for body weight to be maintained longer. The studies can also capture water and food consumption data.

The ALS Therapy Development Institute has used its industrial scale to identify and control for variables in the animal model that may contribute to false-positive or false-negative results. These variables include:

• Mouse litter
• Mouse gender
• Transgene copy number
• Data censoring criteria

Survival efficacy studies at ALS TDI are industrially scaled and adequately powered to identify gender independent or gender specific drug effects.

Because projects at the ALS Therapy Development Institute generate considerable data all the way from therapeutic engineering and production to formulation and in vitro validation to PK/PD to survival efficacy studies, the final results must be analyzed in a comprehensive fashion with input from many scientists and internal research teams. The results of the analysis often spawn new experiments and New Leads.

As a therapeutic moves through the research pipeline at ALS TDI, increasingly more information is collected on its potential efficacy in slowing or stopping ALS. This data package typically consists of information gained from in vitro experiments and in vivo PK, PD and survival efficacy studies. When the research staff believes that this data package would support further advancing the potential therapeutic toward the clinic, it is place in Development. This part of the process is includes the investigation of clinical development options for the specific therapeutic, which is now referred to as a lead candidate.

Every therapeutic developed is unique in several ways, such as the specific pathway(s) it targets, how it has been formulated, etc. These differences become critical components to determining the quickest and most likely successful route to the clinic. Some common clinical development options include: Partner, In-License, Self Generation, Release.

The ALS Therapy Development Institute has a single, profoundly important goal – develop effective therapeutics for ALS as quickly as possible. Currently, ALS TDI does not have a therapeutic in clinical trial, nor is it sponsoring or endorsing any current clinical trials for treating ALS.

In medical research, clinical trials are conducted to allow safety and efficacy data to be collected for potential new therapeutics for ALS. The approval process for a clinical trial requires a great deal of satisfactory information gathered through preclinical research, such as the proper dose range, potential side-effects, mechanism of action, etc. The clinical trial process in the United States is regulated by the Food & Drug Administration (FDA).

The FDA determines a unique and appropriate clinical trial process for each therapeutic. Depending on the type of product and the stage of its development, investigators enroll healthy volunteers and/or patients into small pilot (Phase 1) studies initially to examine tolerability and toxicity issues. However, in certain cases, especially for orphan diseases such as ALS, it is not uncommon for therapeutics to advance directly into a Phase 2 study, which are larger in scale and include actual people living with the disease in order to measure efficacy. Additional trials may be required by the FDA, to replicate or refine any positive safety and efficacy data are gathered. The number of patients is typically increased as the trials advance into the later 3 and 4 phases. Clinical trials can vary in size from a single center in one country to multicenter trials in multiple countries. In is not uncommon for the FDA regulators to require therapeutics to repeat clinical trial phases.

A Project at the ALS Therapy Development Institute is defined as any therapeutic strategy that has been approved by the Research Staff for experimentation. All Projects fall within an existing Research Program or result in the creation of new Research Program. Projects are submitted for discussion and approval during New Lead Meetings that take place on a regular basis at the Institute. Once the project is approved, the therapeutic strategy is advanced through the stages of experimentation and development. These stages include:

• Formulation
• In vitro validation
• In vivo study
• Clinical development

A typical Research Program at the ALS Therapy Development Institute encompasses a broad range of overlapping biological pathways that if modulated by a therapeutic could slow, arrest, or reverse the course of ALS disease progression. A Research Program usually includes the experimental study of more than one treatment strategy directed at one or more specific biological targets. These specific treatment strategies are defined as Projects and are each assigned a Project ID (ex: TDI 0014). ALS TDI currently has Research Program's which employ the following type of therapeutics:

• small molecule drugs
• biologics
• gene therapies
• stem cell therapeutics

Usually, within any Research Program, many experiments using multiple treatment strategies occur in parallel in order to reduce the time necessary to advance the most promising projects forward.

In the cases of small molecule drugs, Formulation includes acquisition or synthesis of the active drug. In most cases, so the drug can be administered in vitro/in vivo, the small molecule drug combined with solvents so that precise doses can be delivered. At the ALS Therapy Development Institute, small molecule drugs can be delivered in vivo using various routes of administration:

• Orally in drinking water
• Orally in food
• Orally by gavage (syringe administration)
• Subcutaneous injection
• Intraperitoneal injection
• Subcutaneous mini-pump implantation
• Intrathecal infusion for delivery of the test compound directly to the spinal cord
• Intracerebral ventricular infusion for delivery of test compound directly to the cerebral spinal fluid around the brain

This class of therapeutics must either be engineered and manufactured internally or acquired from a third party. Unlike small molecule drugs that can be synthesized chemically, biologics generally require biological systems to be generated. Because of this, close attention is directed to ensure the purity, integrity, and biological activity of the therapeutic.

Gene Therapies must be designed and engineered internally prior to being manufactured by third party vendors at a scale adequate to the needs of the rigorous in vitro and in vivo experimentation executed at that ALS Therapy Development Institute. Similar to biologics, gene therapies typically rely on biological systems to be generated. If a biological system is used, the resulting gene therapeutics must be further tested and monitored for purity and effectiveness of the “vector”; the biological or synthetic tool used to deliver the genetic material. The ALS Therapy Development Institute has established relationship with several vector producing organizations, amassing the ability to ramp up several different types of gene therapy products simultaneously.