Potassium channel blockers or derivatives thereof for preventing, alleviating, and / or treating tissue dysfunction caused by toxic injury

Abstract

The present invention provides compositions and methods for preventing, alleviating, or treating a subject having toxicity agent-induced damage. The present invention also provides compositions and methods for preventing, alleviating, or treating tissue damage caused by exposure to a toxicity agent or for restoring at least a portion of normal tissue function. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker. In some embodiments, the potassium channel blocker comprises 4-aminopyridine, a derivative thereof, or a combination thereof. In some embodiments, the pharmaceutical composition can be administered together with an additional therapeutic agent, such as an anticonvulsant.

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims priority to U.S. Provisional Patent Application No. 63 / 367,934, filed on July 8, 2022; U.S. Provisional Patent Application No. 63 / 380,143, filed on October 19, 2022; and U.S. Provisional Patent Application No. 63 / 490,267, filed on March 15, 2023, the contents of each of which are hereby incorporated by reference in their entirety.

Background Art

[0002] Pathological changes caused by exposure to toxic injury can result in a devastating clinical condition for which there is currently no effective treatment. The problems caused by the onset of toxic injury thus represent a major unmet medical need. The injury can occur in many different tissues, with an impact on whether the onset is rapid, delayed, or a combination of the two, and if the condition remains untreated, it may ultimately recover or may ultimately not recover.

[0003] Particularly problematic are toxic injuries that do not occur naturally and require some kind of industrial process to occur. There are several different types of such toxic injuries (e.g., industrial chemicals, chemotherapeutic agents, wartime toxins). The most frequent exposures include non - biological chemicals that did not exist before the development of the chemical industry. The history of the chemical industry dates back thousands of years, but large - scale production of chemicals was first developed in the 19th century. The chemical industry began to emerge in connection with the Industrial Revolution in the late 18th century and began to accelerate in the 19th century when the production of synthetic dyes from coal tar became possible. Since then, the chemical industry has experienced explosive growth in the first half of the 20th century and has continued to grow at an accelerating pace ever since.

[0004] The development of the chemical industry is one of the factors that has exposed people to an increasing number of chemical substances that did not exist in the history of evolution, which is conceptually important in understanding the novelty of the present invention. The fact that these chemical substances were not part of the history of evolution means that there was no evolutionary selection for biological defense strategies. This is a completely different situation from traumatic injuries such as blunt or sharp trauma, or other examples of biological trauma such as myocardial infarction or ischemic injury, or injuries caused by natural biological processes or exposure to toxic chemicals already existing in nature.

[0005] Humans have created tens of millions of chemical substances, most of which have been generated in the past few decades. New substances are isolated or synthesized every few seconds, and it is estimated that recently 10 million new compounds are synthesized in a year. Artificial chemical substances are used for various purposes, and not all of them are toxic, but an important subset of these chemical substances is specially manufactured to utilize their toxic properties.

[0006] For example, chemical substances such as pesticides, avicides, and herbicides are specially developed to have toxic effects on various organisms. Therefore, it is not surprising that such chemical substances can also be harmful to humans. Other chemical substances involved in industrial processes, or developed for the purpose of enabling chemical reactions, or developed for the purpose of generating various types of materials, are incidentally toxic. Other toxic substances such as radiation may be developed for the sterilization of pathogens, cancer treatment, and military use, and have the ability to be intentionally toxic. Exposure to radiation can also incidentally be toxic, for example in the case of accidental exposure.

[0007] Perhaps the most prominent among the chemical substances that cause tissue damage are those that are used to kill cancer cells and are intentionally used on millions of people every year. These chemical substances are specifically produced for the purpose of killing mammalian cells. The desired goal is to selectively target cancer cells, but such selectivity rarely exists, and many normal cells can be damaged by most chemotherapeutic agents. This also applies to new drugs that have been developed to enhance the immune attack against cancer cells but have also been found to exhibit unexpected toxicity. Other examples of such chemical substances are agents developed for agricultural purposes such as pesticides and insecticides, or agents developed for military purposes (such as nerve gases and other incapacitating substances).

[0008] Regarding a specific category of drugs used in the treatment of cancer, an example (among many) of a clinically significant side effect due to exposure to toxic chemicals is chemotherapy-induced peripheral neuropathy (CIPN). In the United States, it is estimated that 30 - 70% of the 650,000 patients who receive chemotherapy each year develop symptoms of CIPN, ranging from numbness and tingling in the hands and feet to burning pain, muscle weakness, and impaired coordination. CIPN is a major cause of medication non-compliance, reduced quality of life, and decreased cancer survival rates. Patients who develop symptomatic CIPN also have an overall treatment cost increase of $20,000 compared to patients who do not develop symptoms and are also at a higher risk of cancer recurrence.

[0009] One of the cancers in which CIPN research is advancing is breast cancer, which was the most commonly seen cancer in women worldwide in 2018 and accounted for more than one - quarter of all cancers. Among the subtypes of breast cancer, hormone - resistant tumors have the worst prognosis. Surgery to remove the tumor and the affected lymph nodes is effective in the early stages of the disease, but the mainstay of treatment for resistant tumors is neoadjuvant chemotherapy. One of the preferred chemotherapeutic agents used in this patient group is taxane, among which paclitaxel (most commonly used in the form of Taxol®) is the most common. PTX is an effective agent in the treatment of hormone - resistant metastatic breast cancer. The problem is that PTX frequently induces CIPN and other toxic side effects on normal tissues (such as nephrotoxicity and damage to the central nervous system). Currently, there is no established treatment for CIPN patients to prevent the onset of CIPN without inhibiting the effect of the chemotherapeutic agent itself or to effectively treat CIPN after its onset (in that case, treatment can refer to treating the symptoms caused by the damage, preventing or repairing the damage, or a combination of any of these features). In pre - clinical studies, multiple compounds that prevent or treat CIPN by blocking ion channels, targeting inflammatory cytokines, and combating oxidative stress have been investigated, but the results are not promising.

[0010] In addition, there are multiple tissues other than the peripheral nerves that are damaged by the therapies used in cancer treatment. For example, chemotherapeutic agents and radiation can damage the heart, lungs, central nervous system, intestines, hair follicles, skin, immune system, cells in the mouth, cells in the genital system, general hematopoietic cells in the bone marrow, and multiple other tissues.

[0011] The lack of an effective treatment for any tissue damage caused by exposure to toxic insults such as chemotherapy, radiation, and / or toxic chemicals represents an as yet unmet medical need. These toxic insults can cause many different types of damage, and existing therapies are usually limited in their application. For example, chelation therapy for exposure to toxic metals (e.g., methyl mercury, lead, etc.) is not useful for treating damage induced by chemotherapy.

[0012] The medical needs represented by tissue damage caused by cancer treatment represent a significant unmet medical need. In many cases, patients face the difficult choice of accepting the potential for toxic reactions to treatment or allowing the cancer to grow without suppression, and damage occurs. In many cases, patients are treated by increasing the dose of chemotherapy agents, radiation, or both until the toxicity reaches an intolerable level. These are referred to as dose-limiting toxicities because they represent the point at which the dose of chemotherapy or radiation necessary to kill cancer cells causes damage to normal tissues at an unacceptable level, causing unacceptable physical symptoms, and thus preventing further escalation of the invasiveness of the anticancer therapy. This may require reducing or completely stopping the cancer treatment dose, and its effectiveness may be limited. Preventing the toxicity caused by the chemicals used in cancer treatment would provide a major advantage in curing cancer, but at present, the ability to provide such an advantage is insufficient to meet the medical need.

[0013] The medical needs represented by tissue damage caused by exposure to harmful chemicals in the environment represent another important unmet medical need. The damage can occur, for example, when an individual is exposed to individual chemicals used for various industrial purposes such as manufacturing or agricultural purposes. Examples of other exposures include, for example, contamination of water sources due to chemical spills or flaring, or other means of exposure in the environment. For example, when toxic substances are generated near a landfill, damage can occur to various tissues. Other examples can include side effects due to exposure to chemicals used for therapeutic purposes other than cancer treatment, for example, side effects due to exposure to fluoroquinolone antibiotics. In these and many other similar types of exposures, due to exposure to various types of toxic injuries generated artificially, an individual can exhibit multiple types of symptoms. If the toxicity caused by such toxic injuries can be prevented, recovered, alleviated, or treated in other ways, it will bring great benefits to medicine. However, at present, the ability to provide such benefits is insufficient for medical needs.

[0014] There are many challenges that hinder the development of treatments for the effects of exposure to toxic injuries. One particularly difficult question is when it is necessary to start treatment. After exposure, is it necessary to start in a short period of time? If so, this may be suitable for situations where the exposure time is known but no opportunity to treat existing damage is provided. In fact, the goal of treating existing damage is very important. Another important issue in the treatment of established damage is whether treatment can provide relief of symptoms that depends on continuous availability of the drug, or whether it can actually provide and promote a permanent recovery that can maintain the effect even after treatment is stopped. The ability to start treatment after the damage and / or symptoms become apparent, relieve the symptoms, and promote permanent recovery, all these goals are important. However, the development of treatments that can provide the benefit of permanent recovery and can be started even after side effects have already appeared is part of the greatest challenges in the development of treatments for the effects of exposure to toxic substances.

[0015] Another type of toxic injury is exposure to high doses of radiation at various locations along the electromagnetic spectrum at an intensity sufficient to cause tissue damage. Examples of such injuries include the heating wavelengths in the infrared region, radiation in the ultraviolet region, and most importantly, radiation toxicity, which represents ionizing radiation with wavelengths of α, β, γ, δ, and θ, that causes damage at the chromosomal and cellular levels.

[0016] Since radiation is used in the treatment of many different types of cancer, the most frequent cause of radiation toxicity is cancer treatment. Radiation can be precisely directed at specific areas of the body, or, up to and including the use of total body irradiation, can be used to irradiate a wider area. Radiation toxicity can also occur in other situations, such as accidents at nuclear power plants, and military situations, such as the use of weapons that emit toxic levels of radiation.

[0017] As already described, damage caused by non-biological chemical substances or radiation toxicity can occur in many different tissues and can manifest in various forms. For example, normal cells that can be damaged by chemotherapeutic agents include, but are not limited to, hematopoietic cells in the bone marrow, hair follicles, cells in the oral cavity, digestive tract, genital system, as well as cells in the heart, kidneys, bladder, salivary glands, auditory system, visual system, lungs, and nervous system. Side effects of treatment with chemotherapeutic agents include, but are not limited to, fatigue, hair loss, easy bruising and bleeding, infections, anemia (decrease in red blood cell count), problems in the mouth, tongue, and throat, such as sores and pain on swallowing, peripheral neuropathy or other nerve problems, such as numbness, tingling, and pain, changes in urine and bladder, as well as kidney problems, cachexia, atrophy of skeletal muscle, and weight changes caused by fibrotic changes, myocardial damage, chemo brain (which can affect concentration and focus), blood vessel damage, problems with reproductive ability (effects on germ cells), anemia, diarrhea, nausea, vomiting, infections, neurological changes (e.g., National Cancer Institute: Side Effects of Chemotherapy Sheet, cancer.gov / cancertopics / coping / physicaleffects / chemo-side-effects).

[0018] In addition to the complexity associated with damage to various tissues, the timing at which toxic side effects are seen can also occur over a wide range of times. Side effects of exposure to toxic injury are classified as acute toxicity (within a few days), early delayed toxicity (within a few weeks), and late delayed toxicity (within months to years). As an example of problems caused by exposure to toxic injury, a subset of pathological outcomes is presented. The selection of these exemplary outcomes is understood to provide non-limiting examples of problems in this general class, and the examples are selected from those that have been particularly well studied.

[0019] An example of the damage caused by toxic injury is a series of pathological conditions included in the broad classification of peripheral neuropathy, which includes CIPN as a subset. The broad class of changes classified as peripheral neuropathy is identified by the presented symptoms such as changes in sensation in the peripheral nerves, but the underlying causes and mechanisms vary widely. For example, some of the common symptoms of peripheral neuropathy occur in various situations, but there is little reason to think that the underlying causes and pathologies are the same, even in neuropathies caused by biological diseases such as diabetic neuropathy, neuropathic pain associated with spinal stenosis, peripheral neuropathy in autoimmune diseases such as Guillain - Barré syndrome, and neuropathies caused by neuropathic pain after spinal cord injury or stroke. The underlying etiologies and pathologies of neuropathies caused by toxic injury represent rather different broad categories of afflictions linked by symptom sharing, rather than being caused by a common mechanism or being treatable by a common approach.

[0020] The defect in using the term peripheral neuropathy as an indicator of a specific pathological process is even more profound in the case of the neuropathy syndromes caused by exposure to toxic injury. Among industrially manufactured chemicals, there are many that can cause symptoms described by the umbrella term peripheral neuropathy in exposed individuals, but the manifestation of such symptoms has little or no relation to the understanding of the etiology, pathophysiology, or treatment of clinical problems. Therefore, the term peripheral neuropathy is used only to indicate that a person is presenting symptoms included in this broad and diverse category, and it should be understood that this term is not associated with a specific etiology or type of injury. There are many types of injuries that result in an outcome collectively called neuropathy, but this does not mean that the detailed nature, etiology, or treatment of the neuropathies is the same.

[0021] Examples of neuropathies involving a wide range of diverse impairments, even when they are caused by exposure to toxic substances, also affect damage to other tissues. Toxic injury can cause various different types of damage to the central nervous system, immune system, gut, heart, lungs, and many other tissues. For example, damage to the central nervous system caused by exposure to chemotherapeutic agents, radiation, or industrial toxins can cause myelin damage, neuron damage, astrocyte activation, promotion of inflammation, as well as various cognitive and neurological symptoms. Given the wide variety of types of damage that occur, it has become apparent that the development of effective treatments for this class of problems is a challenge. The analysis of chemotherapeutic agents is a particularly powerful example of this problem, as the types of damage caused by chemotherapeutic agents are diverse and chemotherapeutic agents can be used to test the hypothesis that certain treatment strategies can be applied to multiple types of damage in multiple tissues.

[0022] Accordingly, there is a need in the art for new compositions and methods for preventing, alleviating, and / or treating tissue dysfunction caused by toxic injury. The present invention meets that need in the art. SUMMARY OF THE INVENTION

[0023] In one aspect, the present invention provides a method for preventing, alleviating, or treating tissue damage or tissue dysfunction caused by toxic injury in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof.

[0024] In one aspect, the present invention provides a method for reducing or restoring tissue damage, oxidative damage, scarring, or any combination thereof caused by toxic injury in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof.

[0025] In one aspect, the present invention provides a method for restoring, improving, or enhancing at least a portion of tissue function reduced by toxic injury in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof.

[0026] In one aspect, the present invention provides a method for restoring, improving, or enhancing demyelination reduced or regulated by toxic injury in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof.

[0027] In one aspect, the present invention provides a method for preventing, alleviating, or treating mitochondrial damage or mitochondrial dysfunction caused by toxic injury in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof.

[0028] In one aspect, the present invention provides a method for preventing, alleviating, or treating axonal injury or axonal dysfunction caused by toxic injury in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof.

[0029] In one aspect, the present invention provides a method for preventing, alleviating, or treating at least one gait disorder caused by toxic injury in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof.

[0030] In one aspect, the present invention provides a method for enhancing tissue regeneration, cell survival, or a combination thereof in a subject in need thereof exposed to at least one toxic injury, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof.

[0031] In various embodiments, the at least one potassium channel blocker comprises 4-aminopyridine, a derivative of 4-aminopyridine, or a combination thereof. In some embodiments, the derivative of 4-aminopyridine is a compound having the structure of formula (I)

Chemical formula

[0032] In some embodiments, the tissue is kidney tissue, liver tissue, heart tissue, lung tissue, brain tissue, central nervous system tissue, peripheral nerve tissue, gastrointestinal tract tissue, intestinal tissue, visual system tissue, auditory system tissue, skin tissue, bladder tissue, reproductive system tissue, hematopoietic system tissue, musculoskeletal tissue, or any combination thereof.

[0033] In some embodiments, the tissue function is motor function, sensory function, cognitive function, visual function, auditory function, renal function, hematopoietic system function, normal skin function, salivary gland function, liver function, gallbladder function, gastrointestinal (GI) function, sexual function, or any combination thereof.

[0034] In some embodiments, the tissue damage is kidney tissue damage, liver tissue damage, heart tissue damage, lung tissue damage, brain tissue damage, central nervous system damage, peripheral nerve tissue damage, peripheral neuropathy, nephropathy, chemotherapy-induced peripheral neuropathy (CIPN), radiation-induced peripheral neuropathy (RIPN), chemotherapy-induced nephrotoxicity (CINT), chemotherapy-induced neutropenia, radiation-induced neutropenia, gastrointestinal tissue damage, intestinal tissue damage, visual system tissue damage, auditory system tissue damage, skin tissue damage, bladder tissue damage, reproductive system tissue damage, hematopoietic system tissue damage, or any combination thereof. In some embodiments, the chemotherapy-induced peripheral neuropathy (CIPN) is CIPN caused by taxanes (P-CIPN), CIPN caused by cisplatin treatment (CisIPN), CIPN caused by anticancer agents, CIPN caused by platinum-based antitumor agents, CIPN caused by vinca alkaloids, CIPN caused by epothilones, CIPN caused by proteasome inhibitors, CIPN caused by immunomodulators, or any combination thereof.

[0035] Accordingly, in one aspect, the present invention provides a method for preventing, alleviating, or treating chemotherapy-induced peripheral neuropathy (CIPN) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof.

[0036] In some embodiments, the tissue damage is multi-tissue damage, multi-organ tissue damage, or any combination thereof.

[0037] In some embodiments, the toxic injury is acute toxic injury, chronic toxic injury, or any combination thereof.

[0038] In some embodiments, treatment can be initiated during the treatment of tissue damage that occurs late, at the onset of injury, during continued exposure to toxic injury, or after exposure to toxic injury has ended.

[0039] In some embodiments, toxic injury includes exposure to radiation at damaging levels along the electromagnetic spectrum.

[0040] In some embodiments, toxic injury is a non-biological substance, non-natural compound, toxin, agent used in the treatment of cancer, chemotherapeutic agent, biological response modifier, radiation, or any combination thereof. In some embodiments, chemotherapeutic agents include platinum-based anti-tumor agents, vinca alkaloid agents, epothilone agents, taxane agents, proteasome inhibitors, immunomodulators, or any combination thereof. In some embodiments, toxic injury includes at least one environmental toxicant. In some embodiments, toxic injury includes at least one industrially produced compound. In some embodiments, radiation includes radiation from radioactive cancer treatment, radiation from nuclear energy accidents, radiation from exposure to nuclear waste, radiation from the use of radioactive materials in military applications, or any combination thereof.

[0041] In some embodiments, toxic injury includes at least a first toxic injury and a second toxic injury. In some embodiments, the first toxic injury is a chemotherapeutic agent, radiation, or a combination thereof, and the second toxic injury is a chemotherapeutic agent, radiation, or a combination thereof.

[0042] In some embodiments, a therapeutically effective amount of a pharmaceutical composition is administered to a subject prior to toxic injury.

[0043] In some embodiments, a therapeutically effective amount of a pharmaceutical composition is administered to a subject at the time of toxic injury.

[0044] In some embodiments, a therapeutically effective amount of a pharmaceutical composition is administered to a subject after toxic injury.

[0045] In some embodiments, a therapeutically effective amount of the pharmaceutical composition is administered to the subject multiple times. In some embodiments, a therapeutically effective amount of the pharmaceutical composition is repeatedly administered to the subject for about 1 day to about 100 years.

[0046] In some embodiments, a therapeutically effective amount of the pharmaceutical composition is administered to the subject systemically, locally, or a combination thereof.

[0047] In some embodiments, a therapeutically effective amount of the pharmaceutical composition is administered to the subject by intraperitoneal injection, intravenous injection, intramuscular injection, intrathecal injection, subcutaneous injection, sublingual administration, inhalation, oral administration, transdermal administration, administration to the outside of the body in the form of a liquid, administration to the outside of the body in the form of an ointment, administration to the outside of the body in the form of a bandage, or any combination thereof.

[0048] In some embodiments, a therapeutically effective amount of the pharmaceutical composition is administered to the subject at a dose of a potassium channel blocker of about 1 mg / day to about 1,000 mg / day. In some embodiments, a therapeutically effective amount of the pharmaceutical composition is administered to the subject at a dose of a potassium channel blocker of about 2.5 mg / day to about 40 mg / day. In some embodiments, a therapeutically effective amount of the pharmaceutical composition is administered to the subject at a dose of a potassium channel blocker of about 40 mg / day to about 100 mg / day.

[0049] In some embodiments, a therapeutically effective amount of the pharmaceutical composition is co-administered with at least one anti-seizure agent or a composition thereof.

[0050] In some embodiments, the at least one anti-seizure agent is a barbiturate, benzodiazepine, bromide, carbamate, carboxamide, fatty acid, fructose or a derivative thereof, gamma-aminobutyric acid (GABA) or an analog thereof, hydantoin, oxazolidinedione, propionate, pyrimidinedione, pyrrolidine, succinimide, sulfonamide, triazine, urea, valproylamide, or any combination thereof.

[0051] In various embodiments, the method further results in enhancing cell survival, reducing scarring, or any combination thereof.

[0052] In various embodiments, the method further results in enhancing the repair or regeneration of endogenous stem cells, enhancing the repair or regeneration of transplanted stem cells, enhancing the repair or regeneration of progenitor cells, promoting neurogenesis, enhancing cell survival, reducing scarring, reducing the size of lesions, reducing oxidative damage, or any combination thereof.

[0053] In one aspect, the invention also provides a method of identifying a subject responsive to 4-aminopyridine administration for preventing, alleviating, or treating tissue damage or tissue dysfunction caused by toxic injury in a subject in need thereof.

[0054] In various embodiments, the method comprises: a) administering to the subject one to five times a therapeutically effective amount of a pharmaceutical composition comprising 4-aminopyridine, a derivative of 4-aminopyridine, or a combination thereof; b) assessing symptoms of tissue damage or tissue dysfunction caused by toxic injury in the subject; and c) identifying that the subject responds to 4-aminopyridine administration for preventing, alleviating, or treating tissue damage or tissue dysfunction caused by toxic injury if symptoms of tissue damage or tissue dysfunction caused by toxic injury in the subject are improved.

[0055] The following detailed description of the preferred embodiments of the invention will be better understood when read in conjunction with the accompanying drawings. For purposes of illustration of the invention, the presently preferred embodiments are shown in the drawings. However, it should be understood that the invention is not limited to the exact construction and means of the embodiments shown in the drawings.

Brief Description of the Drawings

[0056]

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Mode for Carrying Out the Invention

[0057] In the following description, chemotherapy-induced peripheral neuropathy (CIPN) is described as a particularly well-studied non-limiting example of tissue damage caused by exposure to various different types of toxic insults. However, since chemotherapeutic agents can have a wide range of targets and a wide range of molecular structures, lumping some of these toxic substances together as chemotherapeutic agents is itself an over-simplification. Some of these agents, such as taxanes and vinca alkaloids, are closely related to naturally occurring substances, despite being industrially produced for the purpose of treating cancer. In contrast, other agents, such as platinum-containing drugs (e.g., cisplatin and oxaliplatin), are much more similar to industrial chemicals produced for their toxicity.

[0058] One of the best-studied examples of a peripheral neuropathy-like syndrome induced by exposure to toxic chemicals is that induced by treatment with oncology drugs, termed chemotherapy-induced peripheral neuropathy (CIPN) (Hershman et al., 2014, J. Clin. Oncol., 32:1941-1967). CIPN is highly frequent, occurring in 19-85% of treated individuals, with frequencies varying depending on the components of the therapeutic agent (Fallon et al., 2013, Br. J. Anaesth., 111:105-111). Among the factors that modulate the prevalence of CIPN is the specific therapeutic agent used, with reported rates varying from 19% to over 85%, being highest for platinum-based drugs (70-100%), taxanes (11-87%), thalidomide and its analogs (20-60%), and ixabepilone (60-65%) (Banach et al., 2016, Brain Behav., 7:e00558).

[0059] Chemotherapeutic agents can cause various neuropathies, including, for example, large and small fiber, sensory, and / or motor, demyelinating and axonal, cranial and autonomic (e.g., Cioroiu et al., 2017, Curr. Neurol. Neurosci. Rep., 17:47). At a simple level, CIPN is often characterized as a predominantly sensory neuropathy that can often be accompanied by motor and autonomic changes (Seretny et al., 2014, Pain, 155:2461-2470). Additionally, sensations of pain such as spontaneous burning, stabbing or electric shock-like pain, mechanical or thermal allodynia, or hyperalgesia frequently occur (Bernardson et al., 2007, J. Pain Symptom Manag., 34:403-412). In severe cases, CIPN can lead to paralysis, complete immobilization of the patient, and severe physical disability (Mols et al., 2016, Eur. J. Cancer, 69:28-38). Generally, sensory disturbances occur more frequently than autonomic symptoms, which usually include orthostatic hypotension, constipation, and changes in sexual or urinary function (Mols et al., 2016, Eur. J. Cancer, 69:28-38).

[0060] Some compounds such as paclitaxel and oxaliplatin can cause acute neuropathy during or immediately after infusion (Argyriou et al. Clinical pattern and associations of oxaliplatin acute neurotoxicity: A prospective study in 170 patients with colorectal cancer (Cancer. 2013;119:438-444)). In contrast, in many other cases, the symptoms of CIPN can develop weeks or months after the completion of chemotherapy. Some patients experience paradoxical worsening and / or intensification of symptoms after treatment discontinuation (Starbova H et al., 2017, Front. Mol. Neurosci., 10:174). Affected individuals may experience worsening of mild neuropathy or the onset of new CIPN. The severity of symptoms generally correlates with the cumulative dose of the drug (Maestri et al., 2005, Tuori, 91:135-138), but there are also other factors that can influence the severity of symptoms, such as age, genetic factors, exposure to other chemicals or radiation, and various other factors.

[0061] Recent studies have shown that the prevalence of CIPN is as high as approximately 68.1% when measured in the first month after chemotherapy, 60.0% at 3 months, and 30.0% after 6 months (Seretny et al., Pain. 2014;155:2461-2470). Particularly in the case of platinum-based anticancer drugs and taxanes, CIPN can persist for several years after the completion of chemotherapy (Kerckhove et al., Front. Pharmacol. 2017;8:86). The damage that occurs can vary between different chemotherapy compounds, and the compounds that can cause CIPN have each been studied in some detail. Non-limiting examples of the complexity and severity of this problem are shown in the following examples.

[0062] The effects of chemotherapy on the nervous system vary widely among different classes of drugs, depending on the specific physical and chemical properties of the drugs used, as well as their single or cumulative doses (Banach et al., Brain Behav. 2016, 7: e00558). Toxicity can also occur after high single-dose treatments or after cumulative exposure. The symptoms of observed damage to the peripheral nervous system vary widely depending on intensity and duration. They range from acute transient heat sensations to permanent changes in peripheral nerves with chronic pain and irreversible nerve damage (Seretny et al., Pain. 2014, 155: 2461-2470). Six major categories of chemotherapeutic agents damage peripheral sensory neurons, motor neurons, and autonomic neurons, causing CIPN. These are platinum-based antineoplastic drugs (especially oxaliplatin and cisplatin), vinca alkaloids (especially vincristine and vinblastine), epothilones (ixabepilone), taxanes (paclitaxel, docetaxel), proteasome inhibitors (bortezomib), and immunomodulatory drugs (thalidomide) (Starbova H et al., Front. Mol. Neurosci. 2017, 10: 17). Among these agents, the most neurotoxic classes are platinum-based drugs, taxanes, ixabepilone, thalidomide, and their analogs. Vinca alkaloids have low neurotoxicity but can cause multiple other toxicities.

[0063] One of the challenges in developing means to prevent damage caused by chemotherapeutic agents and generally toxic chemicals is that multiple types of disruption occur at the cellular level. Multiple examples of these potential disruptions are shown in the following exemplary embodiments. At a general level, disruptions of normal cellular functions that have been suggested to be associated with the etiology of peripheral nervous system damage include microtubule disruption, oxidative stress, mitochondrial damage, changes in ion channel activity, myelin sheath damage, DNA damage, immune processes, and neuroinflammation. (Zajaczkowska et al., 2019, Int. J. Mol. Sci., 20:1451; ncbi.nlm.nih.gov / pmc / articles / PMC6471666 / ). All of these changes have been suggested to potentially be important in CIPN, but nothing has been shown as to which changes are most important with respect to etiology or treatment. Also, none of these proposals have been useful for the development of either preventive treatments or post-injury therapies, as discussed elsewhere in this application. In short, targeting these changes in a specific way has not led to the development of effective therapeutics, indicating that the discovery of such therapeutics is not easy and not predictable.

[0064] Taxane-induced CIPN provides an example of the complexity of these injuries and their differences from those due to trauma. Taxanes cause CIPN in 11 - 87% of patients (Banach et al., Brain Behav. 2016;7:e00558.). Taxanes have many different effects on cellular function. Paclitaxel is an excellent example of the complexity of the injuries caused by taxanes and other types of toxicants and their differences from those caused by physical trauma. Some of these may be considered direct effects, while others may be indirect effects.

[0065] Taxanes are best known for interfering with microtubule function, which is thought to be associated with the development of CIPN (Gornstein et al., Exp. Neurol. 2017, 288:153 - 166). Taxanes also damage mitochondria, leading to oxidative stress, damage to many cellular components by the production of reactive oxygen species, and axonal transport can be impaired due to these reasons and the effect of taxanes on microtubules. These changes can have multiple effects, such as disruption of signaling, inflammation, and damage to myelin. However, among these changes, myelin damage has been shown to have the least evidence of a causal relationship with CIPN (Areti et al., Redox Biol. 2014;2:289 - 295; Bulua et al., J. Exp. Med. 2011;208:519 - 533; Griffiths et al., J. Pain. 2015;16:981 - 994; Duggett et al., Pain. 2017;158:1499 - 1508). Additional other types of damage associated with paclitaxel treatment include damage to mitochondria, including mitochondrial swelling, vacuolization, and loss of mitochondrial structure (Flatters et al., Pain. 2006;122:245 - 257; Xiao et al., Pain. 2012;153:704 - 709).

[0066] Another possible cause of paclitaxel-induced CIPN is the dysregulation of Ca2+ homeostasis (Siau et al., Anesth. Analg. 2006;102:1485-1490; Yilmaz et al., Cell Calcium. 2017;62:16-28; Kidd et al., J. Biol. Chem. 2002;277:6504-6510; Mironov et al., J. Biol. Chem. 2005;280:715-721). Paclitaxel can cause the release of Ca2+ from mitochondria by opening the mitochondrial permeability transition pore and rapid mitochondrial depolarization, and can also cause the release of Ca2+ from the endoplasmic reticulum. (Kidd et al., J. Biol. Chem. 2002;277:6504-6510; Mironov et al., J. Biol. Chem. 2005;280:715-721; Boehmerle et al., Proc. Natl. Acad. Sci. USA. 2006;103:18356-18361; Li et al., Pain. 2017;158:417-429). Another indicator of the potential contribution of increased Ca2+ levels to CIPN is the increased expression of CaV3.2 channels in paclitaxel-treated rats, and the finding that the suppression of these specific Ca2+ channels restores hypersensitivity (Okubo et al., Neuroscience. 2011;188:148-156). These results indicate that the increase in Ca2+ is an important factor in the development of paclitaxel-induced CIPN. Since one of the properties of 4-AP is to activate high-voltage-activated Ca2+ channels and thus cause Ca2+ release, these studies would conversely indicate the usefulness of 4-AP.

[0067] In paclitaxel-induced CIPN, the expression and function of multiple other ion channels also change. The cation channels TRPV1 and TRPA1, which are important in pain signal transmission, are important in paclitaxel-induced CIPN (Hara et al., Pain. 2013;154:882-889, Materazzi et al., Pflugers Arch. 2012;463:561-569), and TRPA1 antagonists can alleviate the inflammation, cold allodynia, and hyperalgesia induced by paclitaxel (Chen et al., Neuroscience. 2011;193:440-451). Paclitaxel treatment also causes an increase in the number of NaV1.7 channels, and blockade of this channel can attenuate hyperalgesia in rats. (Li et al., J. Neurosci. 2018;38:1124-1136; Aromolaran et al., Mol Pain. 2017;13:1744806917714693; Gheraldin et al., Neuroscience. 2010;169:863-873). Furthermore, a decrease in the expression of K+ channels that cause spontaneous activity of nociceptors was observed in the DRG of a paclitaxel-induced CIPN model (Zhang H et al., Anesthesiology. 2014;120:1463-1475).

[0068] Another cause suggested as an etiology of paclitaxel-induced CIPN is inflammation. Exposure to paclitaxel increases the production of inflammatory cytokines (TNFα and IL-1β), decreases anti-inflammatory cytokines (IL-4 and IL-10), and leads to the attraction and activation of immune cells and the development of neuroinflammation. Paclitaxel can also result in the activation of microglia and astrocytes, as well as an increase in the number of macrophages in the DRG and peripheral nerves. The release of cytokines stimulates the TLR4 receptor within DRG cells, and blocking this receptor reduces pain behavior in mice. IL-10 can also attenuate paclitaxel-induced CIPN. Preventing the development of mechanical allodynia and epidermal nerve fiber loss can also be achieved by suppressing macrophages and microglia (Krukowski et al., J.Neurosci. 2016;36:11074-11083; Zhang et al., J.Pain. 2012;13:293-303; Ruiz-Medina et al., Eur. J.Pain. 2013;17:75-85; Zhang et al., J.Pain. 2016;17:775-786; Liu et al., Mol.Pain. 2010;6:76; Li et al., J.Pain. 2014;15:712-725). Yet another potential cause of paclitaxel-induced CIPN is the activation of astrocytes and the production of inflammatory cytokines associated with effective treatment with minocycline (Zhang et al., J pain. 2012 Mar;13(3):293-303).

[0069] Another drug that disrupts microtubules and causes multiple toxic reactions including CIPN is vincristine, a member of the vinca alkaloid family. Part of the mechanism by which vincristine induces axonal neuropathy is due to disruption of the microtubule axonal transport system. Vincristine binds to tubulin and blocks their polymerization into microtubules. As a result of this activity, vincristine inhibits axonal transport. Vincristine also causes other types of changes in the axonal cytoskeleton (Cioroiu et al., Curr. Neurol. Neurosci. Rep. 2017;17:47). Vincristine induces distal axonal degeneration, and vincristine-induced peripheral neuropathy is associated with pain (Boyette-Davis et al., Pain Manag. 2018;8:363-375). Generally, symptoms of peripheral numbness and tingling begin about 4 to 5 weeks after treatment. Vincristine-induced neuropathy tends to involve both motor and sensory fibers, with small fiber modalities and autonomic fibers being particularly affected (Topp et al., J. Comp. Neurol. 2000;424:563-576).

[0070] The family of platinum-based chemotherapeutic agents is also well known to cause multiple side effects including CIPN. Acute and chronic neurotoxicity after platinum-based chemotherapy are major issues and have contributed to extended infusion times, dose reduction, treatment delays, and even treatment discontinuation (Storey et al., Ann. Oncol. 2010;21:1657-1661). In addition to peripheral neuropathy, cisplatin can also induce ototoxicity, myelotoxicity, and nephrotoxicity.

[0071] Cisplatin is one of the most widely prescribed chemotherapeutic agents and is used in approximately 50% of all cancer chemotherapy regimens. Cisplatin is used in the treatment of a wide range of pediatric and adult malignancies, such as ovarian, testicular, bladder, head and neck, breast, and lung cancers (Galanski et al., Curr. Med. Chem. 2005;12:2075-2094). Cisplatin has multiple toxic side effects, and approximately 40 side effects have been reported to date (Qi et al., Chem. Res. Toxicol. 2019;32:1469-1486). The use of extensive supportive care for cancer patients receiving cisplatin treatment has led to the more common use of high-dose therapy, which includes problems such as acute kidney injury, gastrointestinal disorders such as persistent diarrhea, neuropathy, and hearing loss. Many of the side effects of cisplatin lead to treatment reduction or discontinuation or have a major impact on the quality of life of patients, increasing the level of negative states such as depression and anxiety (Crona et al., Oncologist. 2017;22:609-619, Grunberg et al., Cancer Chemother. Pharmacol. 1989;25:62-64; Van der Hoop et al., Cancer. 1990;66:1697-1702; Perse Cisplatin Mouse Models: Treatment, Toxicity and Translatability Biomedicines. 2021 Oct;9(10):1406). One of the toxic effects of cisplatin treatment is cisplatin-induced peripheral neuropathy (CisIPN), which occurs in a time- and dose-dependent manner. The onset of CysIPN appears to be independent of pretreatment, age, gender, tumor type, and co-treatment with other chemotherapeutic agents (Park et al., Cancer J. Clin. 2013;63:419-437, Schmoll et al., J. Clin. Oncol. 2003;21:4083-4091).

[0072] Furthermore, oxaliplatin treatment can also result in oxaliplatin-induced CIPN. Risk factors for oxaliplatin-induced CIPN, which can present both acutely and chronically, include cumulative oxaliplatin dose, infusion time, low body weight, young age, body surface area >2.0, and several genetic mutations such as mutations in the voltage-gated sodium channel genes SCN4A, SCN9A, and SCN10A (Velasco et al., J. Neurol. Neurosurg. Psychiatry. 2014;85:392-398; Alejandro et al., Am. J. Clin. Oncol. 2013;36:331-337; Palugulla et al., Asian Pac. J. Cancer Prev. 2017;18:3157-3165).

[0073] The anti-tumor mechanisms of platinum-based chemotherapeutic actions may be partially or fully related to CIPN. Other toxicities caused by exposure to these compounds include factors such as DNA binding and the formation of DNA-platinum adducts, resulting in the inhibition of DNA replication and RNA (ribonucleic acid) transcription, followed by the activation of the apoptosis pathway by DNA adducts, the disruption of mitochondrial function and subsequent disruption of the respiratory chain function and increased production of reactive oxygen species (ROS), changes in mitochondrial function and apoptosis activation due to the inhibition of mitochondrial DNA replication and transcription, activation of the immune system (macrophages, T cells, and monocytes) followed by the release of pro-inflammatory cytokines and apoptosis activation, and effects on the calcium signaling pathway and the functions of the protein kinase family (which can also lead to apoptosis) (Dasari et al., Eur. J. Pharmacol. 2014;740:364 - 378; Tesniere et al., Oncogene. 2010;29:482 - 491; Canta et al., Mitochondrial Dysfunction in Chemotherapy-Induced Peripheral Neuropathy (CIPN) Toxics. 2015;3:198 - 223; Ray et al., J. Biomol. Struct. Dyn. 2018 doi:10.1080 / 07391102.2018.1531059; Riddell Cisplatin and Oxaliplatin: Our Current Understanding of Their Actions. Met. Ions Life Sci. 2018 doi:10.1515 / 9783110470734-007; McKeage et al., Br. J. Cancer. 2001;85:1219 - 1225; Sharawy et al., Exp. Toxicol. Pathol. 2015;67:315 - 322; Kober et al., Mol Pain. 2018;14:1 - 16; Jaggi et al., Toxicology. 2012;291:1 - 9; Viatchenko-Karpinski et al., Mol. Pain. 2018;14:1 - 11).In CIPN induced by platinum-containing drugs, although the accumulation of platinum adducts in dorsal root ganglion and trigeminal ganglion neurons has been suggested to be particularly important, considering the breadth of other damaging effects on cell function and the ability of inhibition of other processes to bring about some signs of benefit, other processes also need to be considered for their potential importance.

[0074] Another class of anticancer drugs known to cause other types of damage to CIPN and normal tissues are protease inhibitors, among which bortezomib and carfilzomib have been most studied. Peripheral neuropathy is observed in about one-third of patients treated with these drugs and may persist for weeks, months, or even years after drug discontinuation. Patients receiving bortezomib develop chronic, distal, and symmetric sensory peripheral neuropathy, often accompanied by neuropathic pain syndromes, with the potential for inflammation, myelin damage, and other problems. (Saife et al., J. Peripher. Nerv. Syst. 2010;15:366 - 368; Peng et al., Support. Care Cancer. 2015;23:2813 - 2824; Farquhar-Smith et al., Curr. Opin. Support. Palliat. Care. 2011;5:1 - 7).

[0075] Thus, even in the limited context of CIPN, it is clear that there are multiple types of damage to be addressed regarding damage by toxic injury. Identifying therapeutic agents that can address these multiple types of damage is a difficult scientific challenge, and little has been shown regarding ways to successfully achieve such a goal. The goal of addressing multiple types of damage is contrasted with the more standard goal of treating each type of damage individually, for example, by attempts to identify a treatment targeting mitochondrial health and another targeting the repair of myelin damage (as a non-limiting example).

[0076] Radiation-induced peripheral neuropathy (RIPN) is also a problem in cancer patients. RIPN is often a chronic handicap, progressive, usually irreversible, and often appears several years after radiotherapy. RIPN is less common compared to CIPN but is increasing with the improvement of long-term cancer survival rates. Although the incidence of RIPN has decreased over the past few decades due to technological advancements in RT protocols, if it occurs, it can greatly impair the quality of life of patients (Delanian et al, Radiation-induced neuropathy in cancer survivors, 2012, 105(3):273-282). The pathophysiological mechanism of RIPN is not fully understood. Indirect nerve compression due to extensive radiation-induced fibrosis is thought to play a central role, but other potential factors include axonal injury, demyelination, and vascular damage due to ischemia after capillary network insufficiency. Generally, RIPN is associated with delayed local damage to mature nerve tissue. Delayed effects can include increased damage in irradiated tissue, direct axonal injury and demyelination, extensive fibrosis within and around the nerve trunk, and ischemia due to damage to the capillary network supplying the nerve compensated by angiogenesis (Delanian et al., Radiation-induced neuropathy in cancer survivors, 2012, 105(3):273-282).

[0077] RIPN is thought to be due, in part, to radiation-induced fibrosis (RIF) combined with initial microvascular damage followed by specific neurological damage. RIF itself is a complex process involving multiple elements including fibroblast proliferation, extracellular matrix deposition, amplified by cytokines such as TGFβ1 and CTGF, with inflammation, oxidative stress, damage to the capillary network, and changes in fibroblast function. (Delanian et al., radioher Oncol. 2004;73:119-131, Denham et al., radiother Oncol. 2002;63:129-145). In the acute phase after irradiation, temporary electrophysiological and biochemical changes may occur, combined with changes in vascular permeability of the irradiated nerves (Pradat et al., RevNeurol(Paris). 1994;150:664-677).

[0078] Since radiotherapy is frequently applied locally, a great deal of attention is paid to relatively local types of nerve damage. For example, intracranial nerve damage has been reported after radiotherapy for intracranial and extracranial tumors. Radiation can also cause optic neuropathy, hypoglossal paralysis, facial paralysis, and / or trigeminal neuropathy. When irradiating the upper limb, the treated individual may develop chronic brachial plexopathy, with the time to onset ranging from months to decades after treatment, and the annual average incidence rate being 1.8 - 2.9%, and early manifestation of RIBP being even rarer. In lower limb irradiation, radiation-induced fibrous compression may contribute to damage of the nerve trunk.

[0079] The challenges in the treatment of RIPN are that many problems are generally late-onset, which in itself can cause problems in diagnosis because the contribution of previous radiotherapy cannot be considered (Delanian et al., Radiation-induced neuropathy in cancer survivors, 2012, 105.(3):273-282). The treatment of RIPN is generally symptomatic, and there is a lack of curative strategies. Symptomatic treatments include non-opioid analgesics, benzodiazepines, tricyclic antidepressants, anti-epileptic drugs, and membrane stabilizers (e.g., carbamazepine). Surgical treatment has not been useful, but physical therapy may be useful. The etiology of RIPN initially involves a vascular mechanism, but fibrosis and atrophy are the main targets of treatment intervention. The combination of pentoxifylline-tocopherol (PE) significantly reduces radiation-induced fibrosis due to their synergistic clinical and biological properties (Delanian et al., J Clin Oncol. 2003;21:2545-2550, Hamama et al., radioher Oncol. 2012;105:305-312).

[0080] Clodronate, a bisphosphonate, appears to inhibit osteoclastic bone destruction using an anti-inflammatory effect and inhibit the destruction of rat macrophage myelin nerves (Delanian et al., Semin Radiat Oncol. 2007;17:99-107). Recently, when clodronate was combined with pentoxifylline tocopherol (PENTOCLO), 54 patients with refractory osteoradionecrosis were cured at a median of 9 months (Delanian et al., Int J radiat Oncol Biol Phys. 2011;80:832-839).

[0081] Damage to the nervous system is just one type of damage that can occur as a result of exposure to toxic agents such as chemotherapy, radiation, or industrial chemicals. Although damage to the nervous system has been particularly well studied due to the significant medical consequences of such damage, it is also important to identify treatment strategies that can be used to improve damage caused to other tissues, including but not limited to the kidney, lung, immune system, digestive system, endocrine system, and other tissues specified herein and known to those of ordinary skill in the art.

[0082] For example, the treatment of cancer with chemotherapeutic agents is known to damage the central nervous system and be associated with both neurological and cognitive changes. This is a phenomenon often referred to as "chemobrain" and has been observed in multiple different types of cancer and with multiple different types of chemotherapeutic agents. Changes in cognitive function may be observed during the course of treatment, and damage to the central nervous system can occur rapidly and persist for long periods. Damage to the central nervous system can involve multiple cell types, including myelinating oligodendrocytes, neurons, microglia endothelial cells, and astrocytes. Astrocytes, as the major supporting cells of the brain, appear to be particularly sensitive to such damage and respond by upregulating one of the major cytoskeletal proteins called glial fibrillary acidic protein (GFAP). Such upregulation is thought to be a sign of activation of astrocytes in response to damage.

[0083] Another example of an important and well - studied type of tissue damage caused by exposure to the toxic injury of chemotherapeutic agents is chemotherapy - induced nephrotoxicity (CINT). The damage can occur acutely or persist over a long period (Santos et al., World J Clin Oncol. 2020, 24;11(4):190 - 204; ncbi.nlm.nih.gov / pmc / articles / PMC7186234 / ; Rosner et al., N Engl J Med. 2017;376:1770 - 1781; Perazella et al., Clin J Am Soc Nephrol. 2012;7:1713 - 1721). CINT can be caused by multiple classes of anticancer treatments. As non - limiting examples, alkylating agents can cause acute kidney injury (AKI), hemorrhagic cystitis, inflammatory lesions, syndrome of inappropriate antidiuretic hormone secretion (SIADH), and can damage proximal and distal tubule structures through the action of metabolites and increased cellular oxidative stress. Antimetabolites can cause AKI, decreased glomerular filtration rate (GFR), interstitial edema, and tubular acidosis. Antimicrotubule inhibitors can cause SIADH, while antitumor antibiotics can cause nephrotic syndrome, focal segmental glomerulosclerosis, thrombotic microangiopathy (TMA), AKI, and hemolytic uremic syndrome. Platinum agents can cause AKI, anemia, hypomagnesemia, and proximal urinary tract dysfunction (Rabah et al., Saudi J Biol Sci. 2010;17:105 - 114; Carron et al., Hemodial Int. 2014;18:846 - 847; Shavit et al., Kidney Int. 2014;85:213; Cordonnier et al., Nephrologie. 1985;6:19 - 26; Giroux et al., Am J Kidney Dis. 1985;6:28 - 39; Bitran et al., Cancer. 1982;49:1784 - 1788; Crona et al., Oncologist. 2017;22:609 - 619; Raj et al., J Clin Oncol. 2006;24:3095 - 3100).

[0084] Regarding CIPN, since the causes of CINT are mostly not understood, the development of preventive or therapeutic strategies has become particularly difficult. (Santos et al., World J Clin Oncol. 2020 Apr 24;11(4):190 - 204; ncbi.nlm.nih.gov / pmc / articles / PMC7186234 / ). Similar to other types of toxicities caused by exposure to chemotherapeutic agents, various disorders of renal function have been reported and are included in the general categories of CINT (Santos et al., Nephrotoxicity in cancer treatment: An overview World J Clin Oncol. 2020 Apr 24;11(4):190 - 204; ncbi.nlm.nih.gov / pmc / articles / PMC7186234 / ). For example, treatment can cause AKI due to toxic acute tubular necrosis, TMA, and crystalline nephropathy, proteinuria / nephrotic syndrome due to TMA and glomerular abnormalities, tubular disorders due to electrolyte and acid - base disorders, and chronic kidney disease (CKD) due to glomerular disorders or interstitial nephritis.

[0085] There can also be intrinsic renal injury related to existing patient risk factors mainly associated with advanced age, hypertension, diabetes, congestive heart failure, cirrhosis, liver failure, hyperbilirubinemia, and hypoalbuminemia. There are also kidney - related risk factors such as nephrosis, a history of kidney injury, nephrotic syndrome, and hyper - electrolyte disorders due to vomiting, diarrhea, and the use of diuretics. (Perazella MA, Izzedine H. New drug toxicities in the onco - nephrology world. Kidney Int. 2015;87:909 - 917).

[0086] One of the particularly dangerous side effects of chemotherapy treatment can be the effect of these drugs on the hematopoietic system. Chemotherapy-induced hematotoxicity is a multifactorial problem that affects the treatment of cancer patients. Of these problems, neutropenia (i.e., a decrease in the number of neutrophils) is particularly dangerous because it can increase the risk of infection. However, neutropenia is just one of several hematotoxicities, including thrombocytopenia and anemia (pubmed.ncbi.nlm.nih.gov / 12166034 / ). Bleeding secondary to a decrease in platelets is the major risk posed by chemotherapy-induced thrombocytopenia. The frequency of cancer-related anemia depends on the type of disease, stage, and duration. Chemotherapy-induced anemia is affected by the type of drugs used, the drug administration schedule, and the intensity of the regimen. Fatigue is the most common symptom of anemia and is reported in 80 - 100% of patients receiving chemotherapy. Fatigue is a major factor in the quality of life of patients but is often not systematically and actively treated.

[0087] Despite extensive research on the side effects of chemotherapy agents, there are few drugs that have been proven to be useful, either in preventing or reversing this damage, or in treating the symptoms caused by such damage. Thus, while potential mechanisms of interest have been identified in existing research, these ideas have not been translated successfully, and as such, the results of previous studies cannot serve as predictors of medically useful outcomes.

[0088] This common problem is revealed by examining attempts to develop treatments for CIPN and other types of neurological damage. In this case, extensive research has been done not only on the effects of cancer treatment on the peripheral and central nervous systems. CIPN also has the advantage that it can define assessment criteria that can be measured non-invasively and often obtain quantitative information by using patient questionnaires. Given the high prevalence of CIPN in cancer patients, the improvement in the management of some cancer types, and the increasing number of cancer survivors, discovering new effective strategies to prevent and / or treat CIPN and its long-term consequences is considered an urgent issue. The percentage of cancer patients who experience neuropathic pain at some point during their lifespan ranges from 40% to 70%. However, improved CIPN treatment is needed, and a recent systematic review has shown that there is not enough evidence to confirm the effectiveness of central nervous system (CNS) drugs for CIPN. There are no known preventive methods, and opioids are required for pain control in many patients.

[0089] This problem exists at a general level and, as shown in a recent overview on the toxicity induced by cisplatin (Perse Cisplatin Mouse Models: Treatment, Toxicity and Translatability Biomedicines. 2021 Oct;9(10):1406), it also exists even when looking more specifically at single agents. The authors concluded that "there is no effective therapy to prevent these side effects, and current treatment strategies are symptomatic and of limited effectiveness." Therefore, there is broad agreement that preventing CIPN is a highly desirable goal, and much effort has been made to find agents that can achieve this, but without success (e.g., Hershman et al. Prevention and management of chemotherapy-induced peripheral neuropathy in survivors of adult cancers: American Society of Clinical Oncology clinical practice guideline. J.Clin.Oncol. 2014;32:1941-1967; Hou et al Treatment of chemotherapy-induced peripheral neuropathy: systematic review and recommendations. Pain Physician. 2018;21:571-592). Some compounds were initially promising, but currently there are no compounds that have reached application or are recommended. As described in a recent review on this topic (Zajaczkowska et al Mechanisms of Chemotherapy-Induced Peripheral Neuropathy Int J Mol Sci. 2019 Mar;20(6):1451), "The American Society of Clinical Oncology (ASCO) clinical practice guidelines do not recommend any agents for the prevention of CIPN. For the established treatment of CIPN, duloxetine has been given a moderate recommendation in the ASCO guidelines."There was also a weak recommendation for a topical gel containing baclofen, amitriptyline, and ketamine.

[0090] Looking at this issue in more detail, Cochrane systematic reviews of interventions and systematic reviews by expert groups of the American Society of Clinical Oncology (ASCO) did not recommend the use of a wide range of interventions that had been tried in the field, shown by laboratory research findings, or inferred from results in other fields. Treatments not recommended in this overview included acupuncture, cryotherapy, exercise therapy, ganglioside monosialic acid (GM-1), retinoic acid, amifostine, amitriptyline, calcium magnesium infusion (Ca / Mg), carmagnoside, cannabinoids, carbamazepine, L-carnosine, diethyldithiocarbamic acid (DDTC), gabapentin, pregabalin, glutamic acid, glutathione, Gu-Zhe-Shen-Qi Pills (GJG), metformin minocycline, N-acetylcysteine, nimodipine, omega-3 fatty acids, ORG2766, oxcarbazepine, recombinant human leukemia inhibitory factor, venlafaxine, vitamin B, or vitamin E. Furthermore, acetyl-L-carnitine is not strongly recommended due to high-quality evidence showing worsening of neurotoxic neuropathy. For example, alpha-lipoic acid, OPERA, curcumin, and Neuronorm did not show benefits in randomized controlled trials despite positive findings in preclinical studies. Similarly, despite the success of pilot trials, large-scale phase III trials did not show a significant neuroprotective effect of vitamin E and glutathione supplementation. Patients administered diethyldithiocarbamate (DDTC) with low cumulative doses of cisplatin were also at high risk of discontinuing treatment due to CisPN-related adverse events. Similarly, ORG2766, a hexapeptide analogue of ACTH, increased the incidence of CIPN in smaller cohort studies. Caution is needed with nutritional supplements and supplements whose effectiveness has not been proven. Therefore, previous trials were unable to predict treatment outcomes.

[0091] Furthermore, in two randomized standard-of-care control trials and one non-randomized standard-of-care control trial, glutamine was associated with a reduction in the incidence and severity of sensory impairment, nerve conduction impairment, and impairment of daily function. Amifostine showed clinically meaningful benefits for the prevention of sensory and auditory CIPN but was less likely to be used clinically because of associated worsening of nausea and vomiting. There is also some evidence of the effectiveness of duloxetine in reducing patient discomfort through symptom modification, but again, the average effect is a reduction in pain of 0.72 points on a 0- to 10-point scale.

[0092] In a November 2016 review of this issue by Majithia et al. (New Practical Approaches to Chemotherapy-Induced Neuropathic Pain: Prevention, Assessment, and Treatment. cancernetwork.com / view / new-practical-approaches-chemotherapy)-induced-neuropathic-pain-prevention-assessment-and-treatment), the authors also pointed out that one of the problems in identifying treatments for CIPN is that extrapolation from trials involving patients with other neuropathic pain syndromes must be done carefully because other syndromes may respond to agents to which CIPN does not. Established prevention and treatment of CIPN have generally not been successful. Anticonvulsants such as gabapentinoids and lamotrigine, which are used to treat other neuropathic pain, have not been proven effective for CIPN patients, and symptom severity did not change more than placebo. The level of complexity revealed in the analysis of CIPN is of particular interest because of its prevalence and contribution to the reduction or discontinuation of cancer treatment. Nevertheless, many other cellular and tissue targets affected by cancer therapy are similarly problematic and complex.

[0093] The novelty of the present invention is emphasized by the fact that none of the conventional therapies that are potentially of interest for improving damage caused by chemotherapy and radiation are potassium channel blockers. This applies to CIPN and also to damage in other tissues.

[0094] Another example of the toxicity of cancer therapy treatments and the need for treatments to improve such toxicity involves various types of kidney damage. Kidney damage, including both AKI and CKD, is a common and serious complication of cancer and / or its treatment. AKI is perhaps the most common form of renal insufficiency in cancer patients and has various adverse effects. AKI can also disrupt the bioavailability and / or safety profiles of many oncology drugs, leading to suboptimal treatment, potential for increased risk of drug-induced toxicity. Preventing kidney damage is widely recognized as a means to improve oncological outcomes and prevent unnecessary dose reduction or interruption of oncological treatments that could potentially extend lifespan. Cancer patients who develop AKI in addition to existing CKD also have a higher mortality rate compared to patients without kidney disease.

[0095] Cytotoxic chemotherapy, targeted agents, and immune checkpoint inhibitors are often nephrotoxic. Among cytotoxic chemotherapeutic agents, those most commonly associated with the development of AKI are cisplatin (CDDP), mitomycin C (MM-C), gemcitabine, methotrexate (MTX), ifosfamide, and pemetrexed. Paclitaxel can also cause kidney damage. Cisplatin-induced nephrotoxicity has been particularly well studied and is thought to be multifactorial (Yoo et al Cisplatin nephrotoxicity: a review. Am J Med Sci. 2007, 334:115-124). Cisplatin can cause extensive oxidative stress damage and apoptosis of tubular cells (Volarevic et al. Molecular mechanisms of cisplatin-induced nephrotoxicity: a balance on the knife edge between renoprotection and tumor toxicity. J Biomed Sci 2019;26:25). It can also cause mitochondrial dysfunction, decreased ATPase activity, impaired solute transport, and changes in cation balance. As a result of such changes, sodium and water reabsorption are often decreased, salt and water excretion are increased, and polyuria frequently ensues. In most patients, renal function improves, but some patients develop irreversible renal insufficiency.

[0096] Preventive measures for chemotherapy-induced kidney injury are limited. Excessive fluid replacement and forced diuresis (ultimately the use of mannitol) may reduce the incidence of AKI in patients receiving cisplatin. Intravenous (i.v.) magnesium supplementation (8 - 20 mEq) may also suppress kidney damage, but it is not clear whether this is superior to sufficient oral prehydration with diuresis. (Yamamoto et al. Hydration with 15 mEq magnesium is effective at reducing the risk for cisplatin-induced nephrotoxicity in patients receiving cisplatin(≧50mg / m2)combination chemotherapy. Anticancer Res.2016;36:1873-1877;Saito et al. Premedication with intravenous magnesium has a protective effect against cisplatin-induced nephrotoxicity. Support Care Cancer 2017;25:481-487).

[0097] The only agent approved by the FDA to prevent cumulative nephrotoxicity is amifostine, a ROS scavenger. However, concerns about side effects, cost, and reduced antitumor efficacy limit the use of amifostine in clinical practice. Multiple natural compounds and drugs (e.g., allopurinol and statins) have been proposed to prevent cisplatin- and other cytotoxic agent-related nephrotoxicity, but the level of evidence for all of them appears to be low (Heidari-Soreshjani et al. Phytotherapy of nephrotoxicity-induced by cancer drugs:an updated review. J Nephropathol 2017;6:254-263;Volarevic et al. Molecular mechanisms of cisplatin-induced nephrotoxicity:a balance on the knife edge between renoprotection and tumor toxicity. J Biomed Sci 2019;26:25). For gemcitabine, ifosfamide, and pemetrexed, except for common interventions used to prevent AKI in non-tumor patients, measures to prevent AKI by these agents have not been established at present.

[0098] Speculation continues regarding the etiology of chemotherapy-related thrombotic microangiopathy (TMA), another form of kidney injury caused by cancer treatment. In some cases, microvascular thrombosis is an important event, but it is unclear whether this results from direct endothelial toxicity or from immune-mediated effects on other targets. TMA is also thought to be a common cause of delayed acute kidney injury (AKI) in patients who have undergone hematopoietic stem cell transplantation (HSCT) after high-dose chemotherapy. The etiology of TMA after hematopoietic stem cell transplantation is not fully understood, but renal endothelial cell injury is thought to play an important role (Wanchoo et al. Acute kidney injury in hematopoietic stem cell transplantation. Curr Opin Crit Care 2019;25:531-538).

[0099] Some of the most extensive efforts to discover means of preventing damage caused by exposure to toxic insults have focused on the problem of CIPN. There are several reasons for focusing on this theme, including the large number of people affected by the problem, its importance as a dose-limiting toxicity that can potentially reduce the ability to effectively treat cancer, and the fact that the evolution of CIPN can be studied in a relatively straightforward manner. The only other toxicity that has received a comparable level of attention is chemotherapy-induced hematopoietic damage. However, attempts to develop treatments to overcome the effects of chemotherapy on the hematopoietic system have been far more successful than in the case of CIPN. The classification of CIPN as a peripheral neuropathy has led to a natural interest in determining whether approaches that have shown promise for the treatment of peripheral nerve damage are useful for the prevention or treatment of CIPN. Unfortunately, attempts in this regard have not been successful to date.

[0100] The approach that is the central concern of the present invention, namely the application of potassium channel blockers, preferably 4-AP, for the prevention and / or treatment of CIPN, has not been considered with respect to the problems associated with CIPN. Indeed, upon detailed investigation of the findings regarding 4-AP, significant evidence against the use of 4-AP has emerged even for the narrow purpose of preventing or treating CIPN. There are many reasons indicating that 4-AP is unlikely to be useful for the prevention or treatment of CIPN or other peripheral neuropathies, not to mention the prevention of other types of damage caused by exposure to toxic injury.

[0101] One of the initial suggestions that 4-AP might be useful for pain management was reported in studies involving patients with chronic spinal cord injury. In these studies, 4-AP was reported to reduce neuropathic pain in some individuals (Hansebout et al., 4-Aminopyridine in chronic spinal cord injury: a controlled, double-blind, crossover study in eight patients. J. Neurotrauma 1993;10(1):1-18). Despite multiple additional studies regarding 4-AP and the importance of reducing chronic pain in individuals with spinal cord injury, these studies did not progress to treatment. Instead, a Phase III trial of the effect of 4-AP in individuals with chronic spinal cord injury was unable to demonstrate a significant effect.

[0102] As another example of evidence against the hypothesis that 4-AP is useful for the treatment of CIPN, it was reported in 1988 that 4-AP could alter peripheral nerve conduction properties 3 months after induction of peripheral neuropathy by chronic high-dose exposure to pyridoxine (vitamin B6), a natural compound sometimes used to induce changes in peripheral nerve function (Bowe et al., 1988, Exp. Neurol., 100:448-458). Examination of the details of these studies revealed that they were performed on nerves exposed to 4-AP at a concentration (1 mM) three orders of magnitude higher than the maximum serum concentration considered clinically relevant, and tested in an ex vivo recording chamber at room temperature. Furthermore, it became clear that only the change in nerve sensitivity to 4-AP 3 months after pyridoxine treatment was examined in these studies, and pain analysis was not performed, nor were there any inferences in the paper regarding the understanding or treatment of sensory neuropathy.

[0103] In another study, it was revealed that 4-AP has the ability to promote recovery from acute peripheral nerve traumatic crush injury. As a result of the crush injury being located sufficiently accurately, treatment by local application of a sustained-release formulation of 4-AP, as well as treatment by systemic treatment, was possible (Tseng et al. 4-Aminopyridine promotes functional recovery and remyelination in acute peripheral nerve injury. EMBO Mol Med (2016) 8:1409-1420). Such injuries are qualitatively different from diffuse injuries associated with systemic exposure to chemotherapeutic agents or other toxic insults. In studies using 4-AP to promote recovery from local peripheral nerve crush injuries, it has also been reported that treatment with 4-AP promoted recovery of certain parameters or thermal hyperalgesia, but did not promote recovery of mechanical allodynia. In contrast, in the case of the present invention, 4-AP provides advantages for paclitaxel-induced CIPN with respect to mechanical allodynia, thermal hyperalgesia, and thermal hypoalgesia, which was an abnormal and unexpected outcome due to the opposite direction of symptom change in the response to temperature. Therefore, the different outcomes described above supported the fact that the current outcome could not be predicted from previous results.

[0104] It has also been suggested that electrical stimulation may bring benefits to CIPN patients (Smith et al., Pilot trial of a patient-specific cutaneous electrostimulation device (MC5-A Calmare®) for chemotherapy-induced peripheral neuropathy. J Pain Symptom Manage, 2010, 40:883-891). Despite previous suggestions that electrical stimulation may be useful for the treatment of CIPN, more recent randomized placebo-controlled double-blind trials have shown that "there were no differences between the active drug group and the placebo group with regard to pain, numbness / tingling, symptom frequency, or impact on daily life" (Tonezzer et al. Effects of transcutaneous electrical nerve stimulation on chemotherapy-induced peripheral neuropathy symptoms (CIPN): a preliminary case-control study. J Phys Ther Sci. 2017 Apr;29(4):685-692; ncbi.nlm.nih.gov / pmc / articles / PMC5430273 / ). These results suggest that transcutaneous electrical nerve stimulation applied in frequency-varying mode was unable to improve CIPN symptoms during chemotherapy cycles.

[0105] As another related example, there was no difference in pain scores evaluated by the McGill Pain Questionnaire and the Visual Analog Scale between patients with Guillain-Barre syndrome treated with 4-AP and those treated with placebo (p =.92) (Use of 4-amino pyridine for treatment of peripheral neuropathies (2003) U.S. Patent 6,503,931). Clinically significant changes were not observed in nerve conduction velocity either. (Meythaler et al. Phase IIB Randomized Trial on the Use of 4-Aminopyridine in Guillain-Barre Syndrome. Arch Rehabil Res Clin Transl. 2021 Jun;3(2):100123). Therefore, these studies also showed that 4-AP is not useful for pain treatment with respect to the types of neuropathy that may appear in patients with Guillain-Barre syndrome.

[0106] In this study, we revealed a hitherto unknown and unanticipated method of using the potassium channel blocker 4-AP to prevent and / or treat damage caused by toxic injury. In this study, we investigated the repeated administration of a clinically appropriate dose of 4-AP to individuals exposed to toxic injury. This treatment can be initiated before, during, or after toxic injury. This treatment also has the unexpected effect of being able to prevent damage and promote durable repair at the functional and structural levels. This treatment also has the unexpected effect of rescuing mitochondria from chemotherapy-induced damage. Therefore, this study further provides new findings regarding the novel use of 4-AP to address unmet medical needs. This study demonstrated unexpected advantages that could not be predicted from previous observations. This trial also showed the new finding that 4-AP treatment can be used to treat the effects of harmful exposure to toxic injury, which may include examples such as chemotherapy, radiation, and environmental toxins.

[0107] Furthermore, the present invention provides a novel treatment for any exposure to toxic injury. For example, the present invention provides treatment for tissue damage caused by cancer treatment with chemotherapy and / or radiation. The present invention provides treatment for exposure to toxic substances produced by humans, such as treatment for paclitaxel (PTX)-induced CIPN or cisplatin-induced CIPN (even though these two toxic compounds have different chemical structures and biological activities). The present invention also provides treatment for nephrotoxicity caused by chemotherapy, mitochondrial damage caused by chemotherapy, and effects on the central nervous system caused by chemotherapy.

[0108] Regarding the effects of chemotherapy, in this study, we first focused on CIPN as a specific example of a common problem. Although the mechanisms of damage involved in CIPN are mostly unknown, it is thought to be very different from traumatic injury both in terms of the mechanism of damage and the range of potential damage that can occur. Focusing on the example of paclitaxel-induced CIPN, this is a microtubule-stabilizing agent. Such activity has no obvious relationship with any effects of traumatic injury. A further difference is that the pathological changes that occur after traumatic injury follow a defined order involving local events such as local cell death, immune infiltration, local scarring, and other clearly defined pathological changes. In contrast, exposure to chemotherapy, radiation, or environmental toxins generally does not cause rapid pathological changes and also causes various pathological outcomes that take much longer to develop until onset. Therefore, the novelty of using 4-AP for the treatment of toxic injury lies in the cause of the injury (toxic injury) and the unexpected ability of 4-AP to be useful for the treatment of such injury.

[0109] Accordingly, the present invention is based in part on the unexpected result that 4-AP effectively treated tissue damage caused by toxic injury. Accordingly, the present invention relates in part to compositions and methods for treating a subject exposed to toxic injury such as exposure to a toxic compound or toxic levels of radiation. The method can include administering to the subject a pharmaceutical composition comprising a potassium channel blocker, preferably 4-AP, a derivative thereof, or a combination thereof. In some embodiments, the damage caused by toxic injury can be to multiple sites and / or multiple organs. In some embodiments, the method can include administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising 4-AP, a derivative thereof, or a combination thereof. In some embodiments, the pharmaceutical composition can be formulated to provide a sustained release of 4-AP, a derivative of 4-AP, or a combination thereof. In certain embodiments, 4-AP or a derivative thereof has the formula (I)

Chemical formula

[0110] In some embodiments, 4-AP may be replaced with different potassium channel blockers. 4-AP can bind to multiple potassium channels, and other potassium channel blockers such as 4-AP derivatives and tetraethylammonium can have similar effects in experimental situations. Also, in the case of 4-AP derivatives, 3,4-diaminopyridine is known to have similar effects in some clinical situations. Therefore, in the present invention, it is expected that other potassium channel blockers may also be effective for the purposes for which 4-AP was used in the experimental examples of the present invention.

[0111] In some embodiments, the pharmaceutical composition can be administered to a subject by injection, intraperitoneal injection, intravenous injection, intramuscular injection, intrathecal injection, subcutaneous injection, sublingual administration, inhalation, oral administration, transdermal administration, transplantation, insertion of a device into the subject, or any combination thereof.

[0112] In some embodiments, when the serum level exceeds a defined threshold, the pharmaceutical composition can be administered in combination with an additional therapeutic agent to provide protection against the rare ability of at least one potassium channel blocker to cause seizures in an individual. For example, the pharmaceutical composition can be administered together with an anti-seizure drug. The anti-seizure drug can be selected from lamotrigine, gabapentin, valproic acid, topiramate, famotidine, phenobarbital, diphenylhydantoin, phenytoin, mephenytoin, ethotoin, mephobarbital, primidone, carbamazepine, ethosuximide, methsuximide, phensuximide, trimethadione, benzodiazepine, phenacemide, acetazolamide, progabide, clonazepam, divalproex sodium, magnesium sulfate injection, metaldehyde, paramethadione, sodium phenytoin, sodium valproate, clobazam, sultiam, dilantin, diphenylan, and L-5-hydroxytryptophan, or any combination thereof.

[0113] The pharmaceutical composition may be administered at various times according to the treatment goal. To prevent the effects of toxic injury, prophylactic treatment may be administered before exposure, within one week after exposure, or before the symptoms of the injury become apparent. To slow or reverse the progression of the injury, or to overcome the symptoms of the injury, treatment may be administered at any time, for example, when it is determined that the injury has begun to become apparent due to the manifestation of clinically relevant symptoms.

[0114] If the aim is to prevent the manifestation of injury, the pharmaceutical composition can be repeatedly administered during the period of exposure to toxic injury. In some cases, particularly when treatment is used to alleviate symptoms, treatment can be continued as long as the symptoms persist.

[0115] A subject in need of treatment can be administered 4-AP, a 4-AP derivative, or a combination thereof, at a dosage of about 5 mg / day to about 100 mg / day. In certain embodiments, 4-AP, a 4-AP derivative, or a combination thereof, can be administered to the subject at a dosage of about 5 mg / day to about 40 mg / day, or about 40 mg / day to about 100 mg / day. In some embodiments, the dosage of 4-AP or its derivative can be increased or decreased from these levels according to the effective treatment dosage range from each specific agent.

[0116] The dosage and frequency of administration of a potassium channel blocker or its pharmaceutical composition depend on many factors including, but not limited to, the identity of the potassium channel blocker, the type and severity of the disease or disorder of the subject, the medical condition of the subject, the age of the subject, the sex of the subject, the overall health of the subject, and other factors. It will also be readily apparent to those skilled in the art that appropriate dosages can be determined by clinical trials. Accordingly, the present invention should not be construed as limited to any particular dosing regimen, and the exact dosage and composition to be administered to any subject will be determined by the attending physician, taking into account all other factors about the subject.

[0117] In some embodiments, the methods described herein can be used to treat damage caused by toxic injury in a subject. For example, the methods described herein can be used to treat the effects of exposure to chemotherapy and / or radiation therapy in an individual undergoing cancer treatment. In some embodiments, the methods described herein can be used to treat the effects of exposure to industrial chemicals or toxic radiation due to a nuclear power plant leak, or the effects of the release of toxic radiation in a military scenario. In other embodiments, the methods described herein can be used to treat the effects of exposure to environmental toxicants associated with, for example, air pollution, water pollution, exposure to chemicals released at incinerators, exposure to pesticides, and other environmental exposures well known to those of skill in the art.

[0118] In some embodiments, damage may be most prominent in one system of the body, such as chemotherapy-induced peripheral neuropathy (CIPN). In other embodiments, the damage may appear in multiple tissues, including, for example, the peripheral nervous system, central nervous system, visual system, auditory system, hematopoietic system, gastrointestinal system, bladder, heart, skeletal muscle, hair follicles, skin, vascular system, salivary glands, or any combination of these tissues.

[0119] In some embodiments, the toxic insult can be an industrial chemical used for various purposes such as in agriculture and manufacturing processes, and exposure thereto can cause damage to any of a variety of tissues including, for example, the peripheral nervous system, central nervous system, visual system, auditory system, hematopoietic system, gastrointestinal system, bladder, heart, skeletal muscle, hair follicles, skin, vascular system, salivary glands, or any combination of these tissues. With respect to the present invention, particular focus is placed on agents that are intentionally manufactured by industrial processes and have the ability to cause tissue damage. Whether the agent that causes tissue damage is designed to be intentionally toxic or whether it is accidentally toxic is not the issue. What is important is that the agent can cause tissue damage and / or tissue dysfunction, whether it is in the form of a chemical structure or electromagnetic radiation.

[0120] In some embodiments, the methods described herein can promote the repair and regeneration of nerve cells, such as by restoring at least a portion of the lost motor and / or sensory function of a subject compared to an untreated subject, promoting the generation of nerve cells, increasing cell viability, reducing scarring, or combinations thereof. In further embodiments, the methods described herein can promote the repair and regeneration of nerve cells, such as by restoring at least a portion of the lost motor and / or sensory function of a subject compared to an untreated subject, promoting the generation of nerve cells, increasing cell survival, reducing scarring, reducing other aspects of tissue damage, or combinations thereof. In further embodiments, the methods described herein can promote the repair and regeneration of nerve cells, such as by restoring at least a portion of the lost motor and / or sensory function of a subject compared to an untreated subject, promoting the generation of nerve cells, increasing cell survival, reducing scarring, or combinations thereof.

[0121] In some embodiments, the methods disclosed herein can be used to prevent or treat muscular dystrophy. In some embodiments, muscular dystrophy can result from exposure to chemotherapy, other toxic chemicals, or toxic radiation.

[0122] In some embodiments, the methods disclosed herein can be used to prevent, ameliorate, restore, or otherwise treat tissue dysfunction caused by exposure to artificially generated toxic insults. In some embodiments, tissue dysfunction can occur, for example, in cells of the hematopoietic system, hair follicles, oral cavity, gastrointestinal tract, reproductive system, and cells of the heart, kidney, bladder, salivary gland, auditory system, visual system, lung, nervous system, or any combination thereof.

[0123] In some embodiments, the methods disclosed herein can be used to identify individuals who would benefit from treatment with a potassium channel blocker (e.g., 4-AP, a derivative of 4-AP, or any combination thereof). For example, in some embodiments, the invention relates to a method of identifying a subject responsive to administration of 4-AP for preventing, alleviating, or treating tissue damage or tissue dysfunction caused by toxic injury in a subject in need thereof, the method comprising: a) administering to the subject one to five times a therapeutically effective amount of a pharmaceutical composition comprising 4-AP, a derivative of 4-AP, or a combination thereof; b) assessing symptoms of tissue damage or tissue dysfunction caused by toxic injury in the subject; and c) identifying that the subject responds to administration of 4-AP for preventing, alleviating, or treating tissue damage or tissue dysfunction caused by toxic injury when symptoms of tissue damage or tissue dysfunction caused by toxic injury in the subject improve.

[0124] In some embodiments, an individual exhibiting dysfunction in one or more tissues after exposure to artificially generated toxic injury may be treated with the methods disclosed herein for one to ten days to determine whether tissue function improves with treatment. Thus, in some embodiments, the methods disclosed herein can be used to provide individualized and targeted therapeutics.

[0125] In some embodiments, to prognostically identify individuals who need to continue treatment for a longer period, individuals exposed to a toxic insult and having one or more changes in tissue function caused by the exposure to the toxic insult are treated with 1 to 5 treatments with a potassium channel blocker (e.g., 4-AP, a derivative of 4-AP, or a combination thereof). In some embodiments, if the individual shows improvement in tissue function, the long-term methods disclosed herein are applied. In some embodiments, individuals damaged by a toxic insult and having symptoms in the areas of walking, pain, or nerve conduction velocity are treated with 1 to 5 treatments of 4-AP or a derivative thereof, and the symptoms are measured 1 to 8 hours after the start of treatment (i.e., within 2 times the half-life of 4-AP in serum). For example, in some embodiments, individuals suffering from CIPN or RIPN and having symptoms in the areas of walking, pain, or nerve conduction velocity are treated with 1 to 5 treatments of 4-AP or a derivative thereof, and the symptoms are measured 1 to 8 hours after the start of treatment (i.e., within 2 times the half-life of 4-AP in serum).

[0126] Additional advantages will be described in part in the following description, some will be apparent from the description, or may be learned by practicing the aspects described below. The advantages described below are realized and achieved by the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive.

[0127] Definitions Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are described.

[0128] As used herein, each of the following terms has the meaning associated with it in this section.

[0129] The articles "a" and "an" are used herein to refer to one or more (i.e., at least one) of the grammatical objects of the articles. As an example, "an element" means one element or more than one element.

[0130] As used herein, when referring to measurable values such as amounts, durations, etc., "about" is intended to encompass a variation of ±20%, ±10%, ±5%, ±1%, or ±0.1% from the specified value, when such variation is appropriate for carrying out the methods of the present disclosure.

[0131] As used herein, the term "compound" refers to any specific chemical compound disclosed herein, unless otherwise indicated. In one embodiment, the term also refers to stereoisomers and / or optical isomers (including racemic mixtures) of the disclosed compounds or enantiomerically enriched mixtures.

[0132] As used herein, the term "analog" or "analogue" means a chemical compound or molecule made from a parent compound or molecule by one or more chemical reactions. Thus, an analog may have a structure similar to that of a small molecule therapeutic agent described herein, or may be based on the backbone of a small molecule therapeutic agent described herein, but differ from the small molecule therapeutic agent described herein with respect to certain components or structural configurations that may have a metabolically similar or opposite effect. An analog or derivative can be a small molecule that has a different structure from a reference molecule but retains the essential properties of the reference molecule. An analog or derivative can have a changed interaction with other specific molecules compared to the reference molecule. An analog or derivative molecule can also include salts, adducts, tautomers, isomers, or other variants of the reference molecule.

[0133] The term "derivative" refers to a small molecule that has a different structure from the reference molecule but retains the essential properties of the reference molecule. A derivative may have a different interaction with other specific molecules compared to the reference molecule. A derivative molecule may also include salts, adducts, tautomers, isomers, or other variants of the reference molecule.

[0134] The term "tautomer" refers to a constitutional isomer of an organic compound that is readily interconverted (tautomerizes) by a chemical process.

[0135] The term "isomer" or "stereoisomer" refers to a compound that has the same chemical constitution but differs with respect to the arrangement of atoms or groups in space.

[0136] The term "prodrug" refers to a compound that has a different structure from the reference molecule but is chemically modified by a specific cellular process and is ultimately modified to retain the essential characteristics of the reference molecule or to become the reference molecule. As used herein, the terms "prodrug form" and its derivatives are used to refer to a drug that has been chemically modified to add and / or remove one or more substituents, and when introducing such a prodrug form, such modifications are restored by a naturally occurring process and the drug is reconstituted. Examples of prodrugs include esters, optionally substituted esters, branched esters, optionally substituted branched esters, carbonates, optionally substituted carbonates, carbamates, optionally substituted carbamates, thioesters, optionally substituted thioesters, branched thioesters, optionally substituted branched thioesters, thiocarbonates, optionally substituted thiocarbonates, sulfenylthiocarbonates, optionally substituted sulfenylthiocarbonates, 2-hydroxypropanoic acid esters, optionally substituted 2-hydroxypropanoic acid esters, S-thiocarbonates, optionally substituted S-thiocarbonates, dithiocarbonates, optionally substituted dithiocarbonates, thiocarbamates, optionally substituted thiocarbamates, oxymethoxycarbonyl, optionally substituted oxymethoxycarbonyl, oxymethoxycarbonates, optionally substituted oxymethoxycarbonates, oxymethoxythiocarbonyl, optionally substituted oxymethoxythiocarbonyl, oxymethylcarbonyl, optionally substituted oxymethylcarbonyl, oxymethylthiocarbonyl, optionally substituted oxymethylthiocarbonyl, oxymethoxythiocarbonates, optionally substituted oxymethoxythiocarbonates, L-amino acid esters, D-amino acid esters, oxymethoxyamino esters, N-substituted L-amino acid esters, N,N-disubstituted L-amino acid esters, N-substituted D-amino acid esters, Ν,N-disubstituted D-amino acid esters, sulfenyl, optionally substituted sulfenyl, sulfinyl, sulfonyl, sulfite, sulfate, sulfonamide, imidate, optionally substituted imidate, hydrazonate, optionally substituted hydrazonate, oxymyl, optionally substituted oxymyl, imidininyl, optionally substituted imidininyl, imidyl, optionally substituted imidyl, aminals, optionally substituted aminals, hemiaminals, optionally substituted hemiaminals, acetals, optionally substituted acetals, hemiacetals, optionally substituted hemiacetals, carbonimidates, optionally substituted carbonimidates, thiocarbonimidates, optionally substituted thiocarbonimidates, carbonimidyls, optionally substituted carbonimidyls, carbamimidates, optionally substituted carbamimidates, carbamimidyls, optionally substituted carbamimidyls, thioacetals, optionally substituted thioacetals, S-acyl-2-thioethyls, optionally substituted S-acyl-2-thioethyls, (acyloxybenzyl)ethers, (acyloxybenzyl)esters, PEG esters, PEG carbonates, bis-(acyloxybenzyl)esters, optionally substituted bis-(acyloxybenzyl)esters, (acyloxybenzyl)esters, optionally substituted (acyloxybenzyl)esters, or BAB esters, acetates, formates, and benzoate derivatives of alcohol functional groups in the compound, but are not limited thereto. Methods for structuring a compound as a prodrug can be found in the literature of Testa and Mayer, Hydrolysis in Drug and Prodrug Metabolism, Wiley (2006). Common prodrugs form active metabolites by conversion of the prodrug by hydrolytic enzymes, hydrolysis of amides, lactams, peptides, carboxylic acid esters, epoxides, or cleavage of inorganic acid esters.,

[0137] As used herein, the term "alkyl", by itself or as part of another substituent, unless otherwise indicated, means a straight or branched chain hydrocarbon having the number of carbon atoms indicated (i.e., C1-6 means 1 to 6 carbon atoms). Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, and homologs and isomers such as n-pentyl, n-hexyl, n-heptyl, n-octyl. The term "alkyl" is also intended to include derivatives of alkyl as defined in more detail below, such as "heteroalkyl", "haloalkyl", and "homoalkyl", unless otherwise noted.

[0138] As used herein, the term "substituted alkyl" means an alkyl as defined above, substituted with 1, 2, or 3 substituents selected from halogen, -OH, alkoxy, -NH2, -N(CH3)2, -C(=O)OH, trifluoromethyl, -C≡N, -C(=O)O(C1-C4)alkyl, -C(=O)NH2, -SO2NH2, -C(=NH)NH2, or -NO2, preferably 1 or 2 substituents selected from halogen, -OH, alkoxy, -NH2, trifluoromethyl, -N(CH3)2, or -C(=O)OH, more preferably 1 or 2 substituents selected from halogen, alkoxy or -OH. Examples of substituted alkyl include, but are not limited to, 2,2-difluoropropyl, 2-carboxycyclopentyl, and 3-chloropropyl.

[0139] As used herein, the term "alkylene", by itself or as part of another molecule, means (-CH2-) nAs exemplified by, it means a divalent radical derived from an alkane. As an example, such groups include, but are not limited to, groups having 24 or fewer carbon atoms such as the structures -CH2CH2- and -CH2CH2CH2CH2-. The term "alkylene" is intended to include, unless otherwise indicated, the groups described below as "heteroalkylene".

[0140] As used herein, the terms "alkoxy", "alkylamino", and "alkylthio" are used in their conventional meanings and refer to an alkyl group bonded to a molecule via an oxygen atom, an amino group, and a sulfur atom, respectively.

[0141] As used herein, the term "alkoxy", used alone or in combination with other terms, unless otherwise defined, means an alkyl group having the number of carbon atoms defined and denoted above, bonded to the rest of the molecule via an oxygen atom, and examples thereof include methoxy, ethoxy, 1-propoxy, 2-propoxy (isopropoxy), and higher homologs and isomers. Preferred are (C1-C3) alkoxy, especially ethoxy and methoxy.

[0142] As used herein, the term "halo" or "halogen", used alone or as part of another substituent, unless otherwise defined, means a fluorine, chlorine, bromine, or iodine atom, preferably fluorine, chlorine, or bromine, more preferably fluorine or chlorine.

[0143] As used herein, the term "cycloalkyl" refers to a monocyclic or polycyclic non-aromatic radical in which each of the atoms forming the ring (i.e., the skeletal atoms) is a carbon atom. In one embodiment, the cycloalkyl group is saturated or partially unsaturated. In another embodiment, the cycloalkyl group is fused to an aromatic ring. Cycloalkyl groups include groups having 3 to 10 ring atoms. Specific examples of cycloalkyl groups include, but are not limited to, the following moieties:

Chemical formula

[0144] Monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Bicyclic cycloalkyls include, but are not limited to, tetrahydronaphthyl, indanyl, and tetrahydropentalene. Polycyclic cycloalkyls include adamantane and norbornane. The term "cycloalkyl" includes "unsaturated non-aromatic carbocyclyl" or "non-aromatic unsaturated carbocyclyl" groups, both of which refer to non-aromatic carbocycles as defined herein that contain at least one carbon-carbon double bond or one carbon-carbon triple bond.

[0145] As used herein, the term "heteroalkyl," alone or in combination with other terms, means a stable straight-chain or branched-chain alkyl group consisting of the stated number of carbon atoms and one or two heteroatoms selected from O, N, Si, P, or S, unless otherwise specified, where the nitrogen and sulfur atoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) may be located at any position of the heteroalkyl group, including between the remainder of the heteroalkyl group and a moiety attached thereto, and may be attached to the most distal carbon atom of the heteroalkyl group. Examples include -O-CH2-CH2-CH3, -CH2-CH2-CH2-OH, -CH2-CH2-NH-CH3, -CH2-S-CH2-CH3, and -CH2CH2-S(=O)-CH3. For example, up to two heteroatoms may be consecutive, such as -CH2-NH-OCH3 or -CH2-CH2-S-S-CH3.

[0146] As used herein, the term "heterocycle" or "heterocyclyl" or "heterocyclic", either by itself or as part of another substituent, unless otherwise specified, consists of carbon atoms and at least one heteroatom selected from N, O, or S, and means an unsubstituted or substituted, stable monocyclic or polycyclic heterocyclic ring system in which nitrogen and sulfur heteroatoms are optionally oxidized and nitrogen atoms are optionally quaternized. The heterocyclic system may be attached at any heteroatom or carbon atom that provides a stable structure, unless otherwise specified. The heterocycle may be essentially aromatic or non-aromatic. Examples of 3-membered heterocycloalkyl groups include, but are not limited to, aziridine. Examples of 4-membered heterocycloalkyl groups include, but are not limited to, azetidine and β-lactam. Examples of 5-membered heterocycloalkyl groups include, but are not limited to, pyrrolidine, oxazolidine, and thiazolidinedione. Examples of 6-membered heterocycloalkyl groups include, but are not limited to, piperidine, morpholine, and piperazine. Other non-limiting examples of heterocycloalkyl groups include the following:

Chemical Structure

[0147] Examples of non-aromatic heterocycles include monocyclic groups such as aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, pyrroline, imidazoline, pyrazolidine, dioxolane, sulfolane, 2,3-dihydrofuran, 2,5-dihydrofuran, tetrahydrofuran, thiophene, piperidine, 1,2,3,6-tetrahydropyridine, 1,4-dihydropyridine, piperazine, morpholine, thiomorpholine, pyran, 2,3-dihydropyran, tetrahydropyran, 1,4-dioxane, 1,3-dioxane, homopiperazine, homopiperidine, 1,3-dioxepane, 4,7-dihydro-1,3-dioxepin, and hexamethylene oxide.

[0148] As used herein, the term "aromatic" refers to a carbocyclic or heterocyclic ring containing one or more polyunsaturated rings and having aromatic character, i.e., having (4n+2) delocalized p(π) electrons (where n is an integer).

[0149] As used herein, the term "aryl", used alone or in combination with other terms, unless otherwise defined, means a carbocyclic aromatic system containing one or more rings (generally 1, 2, or 3 rings), such rings may be joined together in a pendant fashion such as in biphenyl or may be fused such as in naphthalene. Examples include phenyl, anthracyl, and naphthyl. Preferred are phenyl and naphthyl, and most preferred is phenyl.

[0150] As used herein, the term "aryl-(C1-C3)alkyl" means a functional group in which a 1-3 carbon alkylene chain is bonded to an aryl group, e.g., -CH2CH2-phenyl. Preferred are aryl-CH2- and aryl-CH(CH3)-. The term "substituted aryl-(C1-C4)alkyl" means an aryl-(C1-C4)alkyl functional group, where the aryl group is substituted. Preferred is substituted aryl(CH2)-. Similarly, the term "heteroaryl-(C1-C4)alkyl" means a functional group in which a 1-3 carbon alkylene chain is bonded to a heteroaryl group, e.g., -CH2CH2-pyridyl. Preferred is heteroaryl-(CH2)-. The term "substituted heteroaryl-(C1-C4)alkyl" means a heteroaryl-(C1-C4)alkyl functional group, where the heteroaryl group is substituted. Preferred is substituted heteroaryl(CH2)-.

[0151] Examples of heteroaryl groups include pyridyl, pyrazinyl, pyrimidinyl (especially 2- and 4-pyrimidinyl), pyridazinyl, thienyl, furyl, pyrrolyl (especially 2-pyrrolyl), imidazolyl, thiazolyl, oxazolyl, pyrazolyl (especially 3- and 5-pyrazolyl), isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,3,4-thiadiazolyl, and 1,3,4-oxadiazolyl.

[0152] Examples of polycyclic heterocycles include indolyl (especially 3-, 4-, 5-, 6-, and 7-indolyl), indolinyl, quinolyl, tetrahydroquinolyl, isoquinolyl (especially 1- and 5-isoquinolyl), 1,2,3,4-tetrahydroisoquinolyl, cinnolinyl, quinoxalinyl (especially 2- and 5-quinoxalinyl), quinazolinyl, phthalazinyl, 1,8-naphthyridinyl, 1,4-benzodioxanyl, coumarin, dihydrocoumarin, 1,5-naphthyridinyl, benzofuryl (especially 3-, 4-, 5-, 6- and 7-benzofuryl), 2,3-dihydrobenzofuryl, 1,2-benzisoxazolyl, benzothienyl (especially 3-, 4-, 5-, 6-, and 7-benzothienyl), benzoxazolyl, benzothiazolyl (especially 2-benzothiazolyl and 5-benzothiazolyl), purinyl, benzimidazolyl (especially 2-benzimidazolyl), benzotriazolyl, thioxanthinyl, carbazolyl, carbolinyl, acridinyl, pyrrolidinizinyl, and quinolizinidinyl.

[0153] The foregoing lists of heterocyclyl and heteroaryl moieties are representative and not limiting.

[0154] As used herein, the term "aminoaryl" refers to an aryl moiety containing an amino moiety. Such amino moieties can include, but are not limited to, primary amines, secondary amines, tertiary amines, masked amines, or protected amines. Such tertiary amines, masked amines, or protected amines may be converted to primary or secondary amine moieties. Further, the amine moiety can include amine-like moieties having chemical properties similar to the amine moiety, including, but not limited to, chemical reactivity.

[0155] As used herein, the term "substituted" means that an atom or group of atoms has replaced hydrogen as a substituent that binds to another group. In the case of an aryl group, an aryl-(C1-C4)alkyl group, and a heterocyclyl group, the term "substituted" as applied to the rings of these groups includes any level of substitution, i.e., mono-, di-, tri-, tetra-, or penta-substitution (when such substitution is permitted). Substituents are independently selected and substitution can occur at any chemically accessible position. In one embodiment, the number of substituents varies between 1 and 4. In another embodiment, the number of substituents varies between 1 and 3. In yet another embodiment, the number of substituents varies between 1 and 2. In yet another embodiment, the substituents are independently selected from 1-6 alkyl, -OH, 1-6 alkoxy, halo, amino, acetamido, or nitro. In yet another embodiment, the substituents are independently selected from 1-6 alkyl, 1-6 alkoxy, halo, acetamido, or nitro. As used herein, the substituent is an alkyl or alkoxy group, and the carbon chain may be branched, straight-chain, or cyclic, with straight-chain being preferred.

[0156] As used herein, the term "optionally substituted" means that the group being referred to may be substituted or unsubstituted. In one embodiment, the group being referred to is optionally substituted with 0 substituents, i.e., the group being referred to is unsubstituted. In another embodiment, the group being referred to is individually optionally substituted with one or more additional groups (s) independently selected from the groups described herein.

[0157] In one embodiment, the substituents are independently oxo, halogen, -CN, -NH2, -OH, -NH(CH3), -N(CH3)2, alkyl (including straight-chain, branched, and / or unsaturated alkyl), substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, fluoroalkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted alkoxy, fluoroalkoxy, -S-alkyl, -S(=O)2alkyl, -C(=O)NH[substituted or unsubstituted alkyl, or substituted or unsubstituted phenyl], -C(=O)N[H or alkyl]2, -OC(=O)N[substituted or unsubstituted alkyl]2, -NHC(=O)NH[substituted or unsubstituted alkyl, or substituted or unsubstituted phenyl], -NHC(=O)alkyl, -N[substituted or unsubstituted alkyl]C(=O)[substituted or unsubstituted alkyl], -NHC(=O)[substituted or unsubstituted alkyl], -C(OH)[substituted or unsubstituted alkyl]2, and -C(NH2)[substituted or unsubstituted alkyl]2. In another embodiment, by way of example, any substituent is selected from oxo, fluorine, chlorine, bromine, iodine, -CN, -NH2, -OH, -NH(CH3), -N(CH3)2, -CH3, -CH2CH3, -CH(CH3)2, -CF3, -CH2CF3, -OCH3, -OCH2CH3, -OCH(CH3)2, -OCF3, -OCH2CF3, -S(=O)2-CH3, -C(=O)NH2, -C(=O)-NHCH3, -NHC(=O)NHCH3, -C(=O)CH3, -ON(O)2, or -C(=O)OH. In yet another embodiment, the substituents are independently C 1-6 alkyl, -OH, C 1-6It is selected from alkoxy, halo, amino, acetamido, oxo, or nitro. In yet another embodiment, the substituent is independently C 1-6 alkyl, C 1-6 selected from alkoxy, halo, acetamido, or nitro. As used herein, the substituent is an alkyl or alkoxy group, and the carbon chain may be branched, straight-chain, or cyclic.

[0158] The term "abnormal" when used with respect to an organism, tissue, cell, or component thereof means that at least one observable or detectable characteristic (e.g., age, treatment, time of day, etc.) is different from that of an organism, tissue, cell, or component thereof that exhibits "normal" (expected) characteristics for each. A characteristic that is normal or expected in one cell or tissue type may be abnormal in another cell or tissue type.

[0159] "Disease" is the health state of an animal impaired by a biologically initiated process. For example, such a biological process can be a pathogen such as a virus or bacterium. They can also be genetic or autoimmune diseases caused by an attack by the immune system on the cells of our own body.

[0160] The term "disorder" generally refers to any impairment of the normal function of the mind or body. For example, the disorders that are the subject of the present invention are those that disrupt the normal state of health and are not diseases as defined in the above description. The present invention focuses on the disruption of normal health and tissue function specifically caused by exposure to artificially generated toxic insults. Non-limiting examples of such toxic insults can be anticancer agents, industrial chemicals, radiation, as well as other disruptors of normal cell and tissue function. There is no single word that represents such disruption. As with all disorders, there are biological consequences. However, in the case of disruption caused by exposure to artificially generated toxic insults, the biological changes occur following the initial exposure to the toxic insult(s).

[0161] As used herein, the terms "cancer" or "neoplasm" include, but are not limited to, benign and malignant cancers of the oral cavity (e.g., mouth, tongue, pharynx, etc.), digestive system (e.g., esophagus, stomach, small intestine, colon, rectum, liver, bile duct, gallbladder, pancreas, etc.), respiratory system (e.g., larynx, lung, bronchus, etc.), bone, joint, skin (e.g., basal cell, squamous cell, melanoma, etc.), breast, genital system (e.g., uterus, ovary, prostate, testis, etc.), urinary system (e.g., bladder, kidney, ureter, etc.), eye, nervous system (e.g., brain), endocrine system (e.g., thyroid, etc.), and hematopoietic system (e.g., lymphoma, myeloma, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, etc.).

[0162] The terms "patient", "subject", "individual", etc. are used interchangeably herein and refer to any animal or its cells suitable for the methods described herein, whether in vitro or in situ. In certain non-limiting embodiments, the patient, subject, or individual is a human.

[0163] As used herein, the term "modulate" means to mediate a detectable increase or decrease in the level of a tissue function or response in a subject as compared to the level of the tissue function or response in the subject in the absence of treatment or compound and / or as compared to the level of the tissue function or response in an untreated but otherwise identical subject. This term encompasses disrupting and / or affecting the original signal or response, thereby mediating an effective therapeutic response in a subject, preferably a human.

[0164] A disease or disorder is "alleviated" if the severity of one or more symptoms or signs of the disease or disorder, the frequency with which such symptoms or signs are experienced by a patient, or both, are reduced.

[0165] As used herein, "treating" a disease or disorder means reducing the frequency or severity of at least one sign or symptom of the disease or disorder experienced by a subject.

[0166] The term "treatment" refers to administering a therapeutically effective amount of a therapeutic agent (e.g., 4-AP) to a subject known or suspected to be exposed to an artificially generated toxic insult.

[0167] The term "therapy" refers to the medical management of a patient intended to cure, ameliorate, stabilize, or prevent a pathologic condition or disorder. This term includes active therapy, i.e., therapy specifically directed toward the improvement of a pathologic condition or disorder. In general, it can include treatment of the cause aimed at removing the cause (i.e., toxic insult) of the associated pathologic condition or disorder. In contrast, the therapy as defined in this application also includes prophylactic therapy, i.e., therapy aimed at minimizing or partially or completely inhibiting the progression of damage that may be caused by exposure to a toxic insult. Such therapy can be used to complement another specific therapy aimed at durable improvement of the associated disease, pathologic condition, or disorder.

[0168] As used herein, "treatment of a disorder of normal health and tissue function specifically caused by exposure to an artificially generated toxic insult" means reducing the severity and / or frequency of signs or symptoms of the disorder of normal health and tissue function experienced by a subject. Such treatment includes prevention of destruction, delay or prevention of progressive worsening by stabilization of destruction, restoration of destruction, and / or partial or complete repair of normal tissue structure and / or function. Treatment can also include managing the symptoms of tissue destruction to reduce its severity, preventing, stabilizing, or restoring dysfunction, and / or partially or completely restoring normal tissue structure and / or function.

[0169] "Treatment for prognostic purposes" refers to using the method of the present invention to identify individuals likely to benefit from continuous treatment. Such treatment for prognostic purposes generally employs short-term treatment, e.g., treatment and analysis over 1 - 5 days, and analyzes any of the symptoms of tissue dysfunction caused by exposure to toxic injury(ies). In this way, responders to treatment can be identified early to further direct attention to the individuals most likely to benefit from the treatment of the present invention.

[0170] As used herein, the terms "treatment" or "treatment regimen" refer to activities carried out to alleviate or modify the disruption of normal health and tissue function, particularly caused by exposure to artificially generated toxic injury, e.g., a series of treatments aimed at reducing or eliminating at least one of the signs or symptoms of such disruption using pharmacological, surgical, dietary, and other techniques. Generally, a treatment regimen may include one or more prescribed doses of drugs or surgery. In the case of the present invention, the treatment regimen may consist of treatment with 4-AP or its derivatives, or different potassium channel blockers, or a combination of such agents. Such treatments may include other agents, such as anticonvulsants, to prevent possible side effects of 4-AP doses in individuals with increased susceptibility to the induction of seizure attacks (seizures are not generally caused by the doses commonly used in the treatment with these agents). Treatment is most often effective and reduces or eliminates at least one sign or symptom of the disruption of normal health and tissue function. In a more general usage of the term "treatment", some examples of the effects of treatment have undesirable effects or side effects, as often occur in cancer treatment, for example. The effects of treatment are also affected by the physiological state of the subject, e.g., age, gender, genetics, weight, the state of other diseases, etc.

[0171] A "therapeutic" treatment is a treatment administered to a subject showing signs of a lesion with the aim of reducing or eliminating those signs.

[0172] The term "nephropathy" refers to a broad category of situations in which kidney function is impaired in some way. With regard to neuropathy, the term "nephropathy" is widely used to describe the symptomatic consequences of various aspects of kidney dysfunction without inferring the cause of such dysfunction. Such tissue dysfunction can occur, for example, as a result of exposure to many different types of toxic injury that are artificially generated, and the cause can be different from nephropathy caused by biological causes such as kidney disease or ischemic reperfusion injury to various tissues that can occur, for example, in myocardial infarction. Nephropathy caused by exposure to artificially generated toxic injury represents another category of unmet medical needs that the present invention addresses.

[0173] One example of the damage caused by toxic injury is a series of pathological conditions included in the broad classification of "peripheral neuropathy." Peripheral neuropathy is a general description of a broad range of changes that are identified by the symptoms that appear, such as changes in the sensation of the peripheral nerves, but the underlying causes and mechanisms are diverse. Thus, the term neuropathy represents the symptoms but includes a wide variety of individual disruptions of normal tissue function. Thus, some of the common symptoms of peripheral neuropathy occur in various situations, but there is little reason to think that the underlying causes and pathologies are the same, even in neuropathies caused by biological diseases, such as diabetic neuropathy, neuropathic pain associated with spinal stenosis, peripheral neuropathy in autoimmune diseases such as Guillain-Barré syndrome, or neuropathies caused by neuropathic pain after spinal cord injury or stroke. The underlying etiologies and pathologies of neuropathies caused by toxic injury represent rather different broad categories of pain linked by the sharing of symptoms, rather than being caused by a common mechanism or being treatable by a common approach. The drawback of using the name peripheral neuropathy as an indicator of a specific pathological process is even more severe in the case of the neuropathy syndromes caused by exposure to toxic injury. Among the industrially manufactured chemicals, there are many that can cause symptoms described by the broad term peripheral neuropathy in exposed individuals, but the manifestation of such symptoms has little or no relation to the understanding of the etiology, pathophysiology, or treatment of clinical problems. Thus, the term peripheral neuropathy is used only to indicate that a person is presenting symptoms included in this broad and diverse category, and it should be understood that this term is not associated with a specific etiology or type of injury. There are many types of injuries that result in an outcome collectively referred to as neuropathy, but this does not mean that the detailed nature, etiology, or treatment of the neuropathy is the same.

[0174] Another example of damage associated with chemotherapy or radiation treatment is damage to the central nervous system. Such damage occurs in the treatment of various different types of cancer, regardless of whether the treatment is directed at the central nervous system. For example, patients undergoing treatment for breast cancer frequently experience cognitive changes associated with various different anticancer treatments and, similarly, often exhibit signs of neurological changes as shown by magnetic resonance imaging studies. As with other types of toxic responses observed in cancer treatment, there is also a lack of treatment strategies for toxic responses in the brain.

[0175] Concerns similar to those described in the two paragraphs above apply to all impairments of normal tissue function, regardless of the tissue in which they occur, i.e., even when specific categorizations of dysfunction are applied, information regarding either the cause or the treatment generally does not become apparent. As a non-limiting example, dysfunction of the visual, auditory, olfactory, respiratory, gastrointestinal, urogenital, musculoskeletal, peripheral nervous, central nervous, musculoskeletal, and other parts of the body can be caused by many different means, and generally, information regarding the cause or treatment does not become apparent from a specific type of dysfunction present in a particular tissue.

[0176] Thus, the observation that tissue dysfunction caused by exposure to artificially generated toxic injury can have characteristics that overlap with tissue dysfunction caused by other causes generally means that using similar terms to classify outcomes into specific functional categories does not imply anything regarding the cause, pathological basis, or treatment of the tissue dysfunction.

[0177] The "effective amount" or "pharmaceutically effective amount" of a compound is an amount of the compound sufficient to produce a beneficial effect in a subject to which it is administered. The "effective amount" of a delivery vehicle is an amount sufficient to effectively bind or deliver the compound. As used herein, the phrase "therapeutically effective amount" refers to an amount sufficient or effective to prevent or treat (delay onset or prevent, prevent progression, inhibit, reduce, or reverse) the disruption of normal health and tissue function caused by exposure to a human-generated toxic insult, including alleviation of symptoms of such disruption.

[0178] A "therapeutically effective amount" refers to an amount that provides a therapeutic effect for a given condition and dosing regimen. In particular, a "therapeutically effective amount" means an amount effective to prevent, alleviate, or improve the symptoms of disruption of normal health and / or tissue function caused by exposure to a human-generated toxic insult, or an amount effective to extend the survival of a subject (human or non-human animal) being treated. Determination of a therapeutically effective amount is within the skill of the art. A "therapeutically effective amount" refers to an amount of a therapeutic agent that, with respect to disruption of normal health and tissue function caused by exposure to a human-generated toxic insult, provides a potentially curative, lasting beneficial effect on the health and well-being of the subject. Beneficial effects on the health and well-being of the subject can include, but are not limited to, (1) cure of a disease state, (2) delay in progression of a disease state, (3) regression of a disease state, (4) reduction of a disease state caused by exposure to a toxic insult. Beneficial effects on the health and well-being of the subject can also include preventive outcomes, which can include (1) preventing or delaying the manifestation of damage to at least one tissue affected by exposure to a toxic insult, (2) maintaining damage at a recovery level after reaching that level by a therapeutically effective amount of a substance, (3) preventing or delaying recurrence of damage after a series of treatments, (4) reducing the likelihood of tissue damage after exposure to a toxic insult, or (5) alleviating any of the symptoms caused by exposure to a toxic insult, but are not limited thereto.

[0179] The terms "pharmacological composition", "therapeutic composition", "therapeutic formulation", or "pharmaceutically acceptable formulation" may mean a composition or formulation that enables effective distribution of the agent provided by the present invention, but is not limited thereto, and the composition or formulation is in a form suitable for administration to the physical site most suitable for its desired action, such as systemic administration. As used herein, the term "pharmaceutical composition" refers to a mixture of at least one compound of the present invention with other chemical components and entities such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients. Pharmaceutical compositions facilitate the administration of compounds to organisms. There are a plurality of techniques for administering compounds in the art, including, but not limited to, intravenous, topical, intraperitoneal, intramuscular, oral, aerosol, parenteral, intraocular, pulmonary, and topical administration. For example, non-limiting examples of agents suitable for formulating with the compounds provided by the present invention include cinnamoyl, PEG, phospholipids or lipophilic moieties, phosphorothioates, P-glycoprotein inhibitors (such as Pluronic P85) that can facilitate the entry of agents into various tissues, such as the CNS (Jolliet-Riant and Tillement, 1999, Fundam. Clin. Pharmacol., 13, 16-26), biodegradable polymers such as poly(DL-lactide-co-glycolide) microspheres for sustained release delivery after transplantation (Emerich, D F et al, 1999, Cell Transplant, 8, 47-58) Alkermes, Inc., Cambridge, Mass., loaded nanoparticles such as polybutylcyanoacrylate nanoparticles that can deliver agents across the blood-brain barrier and alter the uptake mechanism of nerve cells (Prog Neuropsychopharmacol Biol Psychiatry, 23, 941-949, 1999), and oil-based delivery systems (Kirtane et al. 2022, Sci. Adv. 8, eabm8478).

[0180] As used herein, the terms "pharmaceutically acceptable" or "pharmacologically acceptable" can mean, but are not limited to, entities and compositions that do not cause adverse reactions, allergic reactions, or other undesirable reactions when administered to animals or humans as needed. As used herein, "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that are suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reaction, or other problems or complications commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment, in accordance with the guidelines of agencies such as the Food and Drug Administration.

[0181] The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt that can (directly or indirectly) provide the compounds described herein when administered to a subject. Such salts are preferably acid addition salts with physiologically acceptable organic or inorganic acids. Examples of acid addition salts include, for example, mineral acid addition salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphate, and organic acid addition salts such as acetate, trifluoroacetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate, methanesulfonate, and p-toluenesulfonate. Examples of base addition salts include, for example, inorganic salts such as sodium salt, potassium salt, calcium salt, and ammonium salt, and organic base salts such as ethylenediamine, ethanolamine, N,N-dialkyleneethanolamine, triethanolamine, and basic amino acid salts. However, it will be understood that pharmaceutically unacceptable salts may also be useful in the preparation of pharmaceutically acceptable salts and are therefore within the scope of the present invention. Procedures for salt formation are conventional in the art.

[0182] As used herein, the term "pharmaceutically acceptable carrier" refers to a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickening agent, solvent, or encapsulating material, composition, or carrier of a pharmaceutically acceptable material that is involved in transporting or delivering a compound useful in the present invention within or to a subject in order to perform its intended function. Usually, such constructs are transported or delivered from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense that it is compatible with the other components of a formulation containing a compound useful within the present invention and is not harmful to the subject. Some examples of substances that can serve as pharmaceutically acceptable carriers include sugars such as lactose, glucose, and sucrose, starches such as corn starch and potato starch, cellulose, and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate, excipients such as powdered tragacanth, malt, gelatin, talc, cocoa butter, and suppository wax, oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil, glycols such as propylene glycol, polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol, esters such as ethyl oleate and ethyl laurate, buffers such as agar, magnesium hydroxide, and aluminum hydroxide, surfactants, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, phosphate buffer solution, and other non-toxic compatible materials used in pharmaceutical formulations. As used herein, "pharmaceutically acceptable carrier" also includes any coating, antibacterial and antifungal agents, and absorption delaying agents that are compatible with the activity of a compound useful within the present invention and are physiologically acceptable to the subject. Supplementary active compounds can also be incorporated into the composition. "Pharmaceutically acceptable carrier" can further include pharmaceutically acceptable salts of a compound useful within the present invention. Other additional components that can be included in the pharmaceutical compositions used in the practice of the present invention are known in the art.

[0183] The term "pharmaceutically acceptable excipient" generally refers to excipients that are safe, non-toxic and conventionally useful for preparing a desired pharmaceutical composition, and includes excipients acceptable for human pharmaceutical use in addition to veterinary use. Such excipients can be solid, liquid, semi-solid, or in the case of aerosol compositions, gaseous.

[0184] As used herein, the term "solvate" of the present invention means any form in which the active compound in the present invention is bound to another molecule (usually a polar solvent) by non-covalent bonds, and should be understood to particularly include hydrates and alcoholates.

[0185] As used herein, "parenteral administration" of a pharmaceutical composition includes any route of administration characterized by physically breaching the tissue of a subject and administering the pharmaceutical composition through the breach in the tissue. Thus, parenteral administration includes, but is not limited to, administration of a pharmaceutical composition by injection of the composition, application of the composition by surgical incision, application of the composition by non-surgical trauma penetrating the tissue, etc. In particular, parenteral administration is contemplated to include, but is not limited to, intraocular, intravitreal, subcutaneous, intraperitoneal, intramuscular, intradermal, intrasternal injection, intratumoral, intravenous, intraventricular, and renal dialysis infusion techniques.

[0186] Ranges: Throughout this disclosure, various aspects of the invention may be presented in range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Thus, the description of a range should be considered to specifically disclose all the possible sub-ranges and individual numerical values within that range. For example, a description of a range such as 1 to 6 is considered to specifically disclose sub-ranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as the individual numbers within that range, such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the width of the range.

[0187] Description The present invention is based in part on the unexpected result that 4-aminopyridine effectively treats tissue damage caused by toxic injury. Accordingly, the present invention relates in part to compositions and methods for treating a subject exposed to toxic injury, such as exposure to a toxic compound or toxic levels of radiation. The method can include administering to the subject a pharmaceutical composition comprising a potassium channel blocker. In some embodiments, the damage caused by toxic injury can be to multiple sites and / or multiple organs. In some embodiments, the method can include administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising 4-aminopyridine, a derivative thereof, or a combination thereof. In some embodiments, the pharmaceutical composition can be formulated to provide a sustained release of 4-aminopyridine, a derivative of 4-aminopyridine, or a combination thereof. In certain embodiments, 4-aminopyridine or a derivative thereof has the formula (I) [Chemical Formula] or can be represented by the structure of an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof. In one embodiment, R 1 , R 2 , R 3 , R 4 , and R 5 are each independently selected from hydrogen, halogen, C1-C6 alkyl, amine, hydroxyl, alkoxy, carboxyl, or any combination thereof, wherein R 1 , R 2 , R 3 , R 4 , and R 5 are optionally substituted. In some embodiments, the derivative of 4-aminopyridine includes 3,4-diaminopyridine, 3-hydroxy-4-aminopyridine, or a combination thereof.

[0188] In some embodiments, the methods described herein can be used to treat damage caused by toxic injury in a subject. For example, the methods described herein can be used to treat the effects of exposure to chemotherapy or other chemical cancer treatments and / or radiation therapy in an individual undergoing cancer treatment. In some embodiments, the methods described herein are used to treat the effects of exposure to industrial chemicals due to a nuclear power plant leak, chemicals produced from various chemicals by a chemical reaction (e.g., those that can occur in an incinerator), or exposure to toxic radiation, or the effects of the release of toxic radiation in a military scenario.

[0189] In some embodiments, damage can be most prominent in one system of the body, such as chemotherapy-induced peripheral neuropathy. In other embodiments, the damage can appear in multiple tissues, including, for example, the peripheral nervous system, central nervous system, visual system, auditory system, hematopoietic system, gastrointestinal system, bladder, urogenital system, heart, skeletal muscle, hair follicles, skin, vascular system, salivary glands, or any combination of these tissues.

[0190] In some embodiments, the methods described herein can promote nerve cell repair and regeneration, such as promoting the generation of nerve cells, enhancing cell survival rate, reducing scarring, or combinations thereof, by restoring at least a portion of the lost motor and / or sensory functions of the subject as compared to an untreated subject. In further embodiments, the methods described herein can promote nerve cell repair and regeneration, such as promoting the generation of nerve cells, enhancing cell survival, reducing scarring, reducing other aspects of tissue damage, or combinations thereof, by restoring at least a portion of the lost motor and / or sensory functions of the subject as compared to an untreated subject. In further embodiments, the methods described herein can promote nerve cell repair and regeneration, such as promoting the generation of nerve cells, enhancing cell survival, reducing scarring, or combinations thereof, by restoring at least a portion of the lost motor and / or sensory functions of the subject as compared to an untreated subject.

[0191] In some embodiments, the methods disclosed herein can be used to prevent or treat muscular dystrophy. In some embodiments, muscular dystrophy can be caused by chemotherapy, other toxic chemicals, or treatment with toxic radiation.

[0192] In other embodiments, the methods disclosed herein can be used to prevent or treat dysfunction in the peripheral nervous system, central nervous system, visual system, auditory system, hematopoietic system, gastrointestinal system, bladder, urogenital system, heart, skeletal muscle, hair follicles, skin, vascular system, salivary glands, or any combination thereof. In some embodiments, tissue dysfunction can be caused by chemotherapy treatment, or exposure to other toxic chemicals or toxic radiation.

[0193] Compounds and Compositions In one aspect, the present invention provides compounds effective to prevent, mitigate, and / or treat tissue damage caused by toxic injury, tissue dysfunction caused by toxic injury, mitochondrial dysfunction caused by toxic injury, muscular atrophy caused by toxic injury, neuropathy caused by toxic injury, or other types of dysfunction in other aspects of the body, or any combination thereof. By way of non-limiting example, such dysfunctions can include dysfunctions of the visual system, auditory system, olfactory system, respiratory system, gastrointestinal system, urogenital system, musculoskeletal system, peripheral nervous system, central nervous system, musculoskeletal system, and dysfunctions in other parts of the body can be caused by many different means, and specific types of dysfunctions are present in specific tissues.

[0194] In one aspect, the present invention provides a compound effective to reduce or restore tissue damage caused by toxic injury, oxidative damage caused by toxic injury, and / or scarring caused by toxic injury. In one aspect, the present invention provides a compound effective to restore, improve, and / or enhance at least a portion of a tissue function affected by toxic injury, myelination affected by toxic injury, tissue regeneration affected by toxic injury, cell survival affected by toxic injury, cell generation affected by toxic injury, repair or regeneration of endogenous stem cells or progenitor cells affected by toxic injury, repair or regeneration of transplanted stem cells or progenitor cells affected by toxic injury, repair or regeneration of stem cells or progenitor cells affected by toxic insult, or any combination thereof. In one aspect, the present invention provides a compound effective to reduce the size of a lesion caused by toxic injury, or any combination thereof. In one aspect, the present invention provides a compound effective to inhibit at least one ion channel affected by 4-AP or a derivative thereof.

[0195] Accordingly, in some embodiments, the compound is a potassium channel blocker having the structure of formula (I), or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof.

Chemical formula

[0196] In various embodiments, R 1is hydrogen, deuterium, halogen, alkyl, cycloalkyl, heterocycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxycarbonyl, amino, aminoalkyl, aminoaryl, aminoalkyl-aryl, aminoheteroaryl, aminoalkyl-heteroaryl, amide, aminoalkenyl, aminoalkynyl, aminoacetate, acyl, hydroxyl, hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxyaryl, alkoxy, carboxyl, carboxylate, ester, =O, -NO2, -CN, sulfoxy, sulfonyl, alkylsulfonyl, secondary amide, tertiary amide, amino acid, or any combination thereof. In some embodiments, R 1 is optionally substituted. For example, in some embodiments, R 1 is hydrogen, halogen, C1-C6 alkyl, amine, hydroxyl, alkoxy, carboxyl, or any combination thereof.

[0197] In various embodiments, R 2 is hydrogen, deuterium, halogen, alkyl, cycloalkyl, heterocycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxycarbonyl, amino, aminoalkyl, aminoaryl, aminoalkyl-aryl, aminoheteroaryl, aminoalkyl-heteroaryl, amide, aminoalkenyl, aminoalkynyl, aminoacetate, acyl, hydroxyl, hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxyaryl, alkoxy, carboxyl, carboxylate, ester, =O, -NO2, -CN, sulfoxy, sulfonyl, alkylsulfonyl, secondary amide, tertiary amide, amino acid, or any combination thereof. In some embodiments, R 2 is optionally substituted. For example, in some embodiments, R 2is hydrogen, halogen, C1-C6 alkyl, amine, hydroxyl, alkoxy, carboxyl, or any combination thereof.

[0198] In various embodiments, R 3 is hydrogen, deuterium, halogen, alkyl, cycloalkyl, heterocycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxycarbonyl, amino, aminoalkyl, aminoaryl, aminoalkyl-aryl, aminoheteroaryl, aminoalkyl-heteroaryl, amide, aminoalkenyl, aminoalkynyl, aminoacetate, acyl, hydroxyl, hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxyaryl, alkoxy, carboxyl, carboxylate, ester, =O, -NO2, -CN, sulfoxy, sulfonyl, alkylsulfonyl, secondary amide, tertiary amide, amino acid, or any combination thereof. In some embodiments, R 3 is optionally substituted. For example, in some embodiments, R 3 is hydrogen, halogen, C1-C6 alkyl, amine, hydroxyl, alkoxy, carboxyl, or any combination thereof.

[0199] In various embodiments, R 4is hydrogen, deuterium, halogen, alkyl, cycloalkyl, heterocycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxycarbonyl, amino, aminoalkyl, aminoaryl, aminoalkyl-aryl, aminoheteroaryl, aminoalkyl-heteroaryl, amide, aminoalkenyl, aminoalkynyl, aminoacetate, acyl, hydroxyl, hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxyaryl, alkoxy, carboxyl, carboxylate, ester, =O, -NO2, -CN, sulfoxy, sulfonyl, alkylsulfonyl, secondary amide, tertiary amide, amino acid, or any combination thereof. In some embodiments, R 4 is optionally substituted. For example, in some embodiments, R 4 is hydrogen, halogen, C1-C6 alkyl, amine, hydroxyl, alkoxy, carboxyl, or any combination thereof.

[0200] In various embodiments, R 5 is hydrogen, deuterium, halogen, alkyl, cycloalkyl, heterocycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxycarbonyl, amino, aminoalkyl, aminoaryl, aminoalkyl-aryl, aminoheteroaryl, aminoalkyl-heteroaryl, amide, aminoalkenyl, aminoalkynyl, aminoacetate, acyl, hydroxyl, hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxyaryl, alkoxy, carboxyl, carboxylate, ester, =O, -NO2, -CN, sulfoxy, sulfonyl, alkylsulfonyl, secondary amide, tertiary amide, amino acid, or any combination thereof. In some embodiments, R 5 is optionally substituted. For example, in some embodiments, R 5is hydrogen, halogen, C1-C6 alkyl, amine, hydroxyl, alkoxy, carboxyl, or any combination thereof.

[0201] For example, in one embodiment, the compound represented by formula (I) is 4-AP, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof, 3,4-diaminopyridine, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof, 3-hydroxy-4-aminopyridine, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof, N-(4-pyridyl)-t-butylcarbamate, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof, N-(4-pyridyl)ethylcarbamate, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof, N-(4-pyridyl)methylcarbamate, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof, N-(4-pyridyl)isopropylcarbamate, or an analog, racemate, tautomer, isomer, enantiomer, diastereomer, prodrug, pharmaceutically acceptable salt, or derivative thereof, or any combination thereof.

[0202] The compounds described herein can form salts with acids or bases, and such salts are included in the present invention. The term "salt" encompasses addition salts of the free acid or free base that are compounds of the present invention.

[0203] In one aspect, the present invention relates in part to a composition comprising one or more compounds of the present invention. In some embodiments, the composition comprises one or more compounds having the structure of formula (I), or analogs, racemates, tautomers, isomers, enantiomers, diastereomers, prodrugs, pharmaceutically acceptable salts, or derivatives thereof. In some embodiments, the composition is a pharmaceutical composition. Thus, in one aspect, the present invention provides a composition effective to prevent, mitigate, and / or treat tissue damage caused by toxic injury, tissue dysfunction caused by toxic injury, mitochondrial dysfunction caused by toxic injury, muscle atrophy caused by toxic injury, neuropathy caused by toxic injury, or any combination thereof.

[0204] In one aspect, the present invention provides a composition effective to reduce or reverse tissue damage or dysfunction caused by toxic injury, oxidative damage caused by toxic injury, and / or scarring caused by toxic injury. In one aspect, the present invention provides a composition effective to restore, improve, and / or enhance at least a portion of tissue function affected by toxic injury, myelination affected by toxic injury, tissue regeneration affected by toxic injury, cell survival affected by toxic injury, neurogenesis affected by toxic injury, repair or regeneration of endogenous stem or progenitor cells affected by toxic injury, repair or regeneration of transplanted stem or progenitor cells affected by toxic injury, repair or regeneration of stem or progenitor cells affected by toxic injury, or other types of functional injury in other aspects of the body, or any combination thereof. By way of non-limiting example, such dysfunctions can include dysfunctions of the visual system, auditory system, olfactory system, respiratory system, gastrointestinal system, urogenital system, musculoskeletal system, peripheral nervous system, central nervous system, musculoskeletal system, and / or other parts of the body. Tissue dysfunction can be caused by a variety of different means. Further, the presence of a particular type of dysfunction in a particular tissue does not distinguish between the various types of damage that can result in a symptomatically similar outcome, as in the case of peripheral neuropathy. In one aspect, the present invention also provides a composition effective to reduce the lesion size affected by toxic injury.

[0205] In some embodiments, the toxic injury is an acute toxic injury, a chronic toxic injury, or a combination thereof. Examples of such toxic injuries include non-biological substances such as chemotherapeutic agents, naturally occurring toxic non-biological substances (e.g., arsenic, lead, mercury), non-natural compounds, toxins, environmental poisons, drugs used in cancer treatment (e.g., chemotherapeutic agents), biological response modifiers, toxic industrial chemicals (e.g., chemicals used in manufacturing or agricultural settings), radiation (e.g., cancer treatment, industrial accidents, military exposure), naturally occurring toxic radiation (e.g., harmful amounts of heat generated by fire, extreme weather conditions, and / or exposure to sunlight-related ultraviolet light sufficient to cause tissue damage), or combinations thereof, but are not limited thereto.

[0206] In various embodiments, the toxic injury is an anti-cancer agent. The anti-cancer agent can be any anti-cancer agent known in the art. In certain embodiments, the anti-cancer agent can be effective in the treatment of one or more of pancreatic cancer, esophageal cancer, rectal cancer, colon cancer, prostate cancer, kidney cancer, liver cancer, breast cancer, ovarian cancer, and gastric cancer. Examples of anti-cancer agents include, but are not limited to, chemotherapeutic agents, anti-proliferative agents, anti-tumor agents, checkpoint inhibitors, and anti-angiogenic agents. For example, in one embodiment, the anti-cancer agent is gemcitabine, doxorubicin, 5-FU, a tyrosine kinase inhibitor, sorafenib, trametinib, rapamycin, fulvestrant, enzalutamide, or paclitaxel.

[0207] Examples of chemotherapeutic agents include cytotoxic agents (e.g., 5-fluorouracil, cisplatin, carboplatin, methotrexate, daunorubicin, doxorubicin, vincristine, vinblastine, oxorubicin, carmustine (BCNU), lomustine (CCNU), cytarabine USP, cyclophosphamide, estramustine phosphate sodium, altretamine, hydroxyurea, ifosfamide, procarbazine, mitomycin, busulfan, cyclophosphamide, mitoxantrone, carboplatin, cisplatin, recombinant interferon α-2a, paclitaxel, teniposide, and streptozocin), cytotoxic alkylating agents (e.g., busulfan, chlorambucil, cyclophosphamide, melphalan, or ethyl sulfonate), alkylating agents (e.g., azaleic acid, AZQ, BCNU, busulfan, bisulfan, carboxyphthalato platinum, CBDCA, CCNU, CHIP, chlorambucil, chloroazotocin, cisplatin, chromeson, cyanomorpholino doxorubicin, cyclodizone, cyclophosphamide, dianhydrogalactitol, fluorodopan, hepsulfam, hikanton, ifosfamide, melphalan, methyl CCNU, mitomycin C, mitozolomide, nitrogen mustard, PCNU, piperazine, piperazinedione, pipobroman, porfiromycin, spirohydantoin mustard, streptozotocin, teloxiron, tetraplatin, thiotepa, triethylenemelamine, uracil nitrogen mustard, Yoshi-864), antimitotic agents (e.g., allocolchicine, halicondrin M, colchicine), colchicine derivatives, dolastatin 10, maytansine, lysocine, paclitaxel derivatives, paclitaxel, thiocolchicine, tritylcysteine, vinblastine sulfate, and vincristine sulfate), plant alkaloids (e.g., actinomycin D, bleomycin, L-asparaginase, idarubicin, vinblastine sulfate, vincristine sulfate, mitramycin, mitomycin, daunorubicin, VP-16-213, VM-26, navelbine, and taxotere), biological agents (e.g., α interferon, BCG, G-CSF, GM-CSF, and interleukin-2).Topoisomerase I inhibitors (e.g., camptothecin, camptothecin derivatives, morpholinodoxorubicin), topoisomerase II inhibitors (e.g., mitoxantrone, amonafide, m-AMSA, anthrapyrazole derivatives, pyrazoloacridine, bisantrene HCL, daunorubicin, doxorubicin, menogaril, N,N-dibenzyl daunomycin, oxantrazole, rubidazone, VM-26, and VP-16), and compounds (e.g., hydroxyurea, procarbazine, o,p’-DDD, dacarbazine, CCNU, BCNU, cis-diaminedichloroplatinum, mitoxantrone, CBDCA, levamisole, hexamethylmelamine, all-trans retinoic acid, Gliadel, and porfimer sodium), or combinations thereof, but not limited to these.

[0208] An antiproliferative agent is a compound that reduces cell proliferation. Examples of antiproliferative agents include alkylating agents, antimetabolites, enzymes, biological response modifiers, other drugs, hormones, and antagonists, androgen inhibitors (e.g., flutamide and leuprolide acetate), antiestrogens (e.g., tamoxifen citrate and its analogs, toremifene, droloxifene, and raloxifene), levamisole, gallium nitrate, granisetron, strontium-89 chloride, filgrastim, pilocarpine, dexrazoxane, ondansetron, or any combination thereof, but not limited to these.

[0209] Examples of anti-tumor agents include, but are not limited to, cytotoxic / anti-tumor agents and anti-angiogenic agents. Cytotoxic / anti-tumor agents are defined as agents that attack and kill cancer cells. Some cytotoxic / anti-tumor agents are alkylating agents such as, for example, cis-platin, cyclophosphamide, nitrogen mustard, trimethiophosphoramide, carmustine, busulfan, chlorambucil, bendamustine, uracil mustard, chromafazine, and dacarbazine, which alkylate genetic material within tumor cells. Other cytotoxic / anti-tumor agents are antimetabolites of tumor cells, such as cytosine arabinoside, fluorouracil, methotrexate, mercaptopurine, azathioprine, and procarbazine. Other cytotoxic / anti-tumor agents are antibiotics such as doxorubicin, bleomycin, dactinomycin, daunorubicin, mitramycin, mitomycin, mitomycin C, and daunomycin. There are numerous commercially available liposomal formulations of these compounds. Still other cytotoxic / anti-tumor agents are mitotic inhibitors (vinca alkaloids). These include vincristine, vinblastine, and etoposide. Various cytotoxic / anti-tumor agents include taxol and its derivatives, L-asparaginase, anti-tumor antibodies, dacarbazine, azacitidine, amsacrine, melphalan, VM-26, ifosfamide, mitoxantrone, and vindesine.

[0210] Anti-angiogenic agents are well-known to those skilled in the art. Examples of anti-angiogenic agents include, but are not limited to, anti-VEGF antibodies (including humanized and chimeric antibodies), anti-VEGF aptamers, antisense oligonucleotides, angiostatin, endostatin, interferon, interleukin 1 (including α and β), interleukin 12, retinoic acid, and tissue inhibitors of metalloproteinases -1 and -2 (TIMP-1 and -2), small molecules including topoisomerase such as razoxane, topoisomerase II inhibitors having anti-angiogenic activity, or any combination thereof.

[0211] Examples of other anti-cancer agents include acivicin, aclarubicin, acodazole hydrochloride, acronine, adozelesin, aldosterone, altretamine, ambomycin, ametantrone acetate, aminoglutethimide, amsacrine, anastrozole, anthramycin, asparaginase, asperlin, azacitidine, azetepa, azotomycin, batimastat, benzodepa, bicalutamide, bisantrene hydrochloride, bisnafide dimesylate, bizelesin, bleomycin sulfate, brequinar sodium, broxuridine, busulfan, caracemide, carbetimer, carboplatin, carmustine, carboquone hydrochloride, carzelesin, cedefingol, chlorambucil, cirolemycin, cisplatin, cladribine, crisnatol mesylate, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin hydrochloride, decitabine, dexormaplatin, dezaguanine, dezaguanine mesylate, diaziquone, docetaxel, doxorubicin, doxorubicin hydrochloride, droloxifene, droloxifene citrate, drostanolone propionate, duazomycin, edatrexate, eflornithine hydrochloride, elsamitrucin, enloplatin, enpromate, epipropidine, epirubicin hydrochloride, erbuzole, esorubicin hydrochloride, estramustine, estramustine sodium phosphate, etanidazole, etoposide, etoposide phosphate etoprine, fadrozole hydrochloride, fazarabine, fenretinide, floxuridine, fludarabine phosphate, fluorouracil, fluorocytosine, fosquidone, fostriecin sodium, gemcitabine, gemcitabine hydrochloride, hydroxyurea, idarubicin hydrochloride, ifosfamide, ilmofosine, interleukin II (including recombinant interleukin II, or rIL2), interferon α-2a, interferon α-2b, interferon α-n1, interferon α-n3, interferon β-Ia, interferon γ-Ib, iplatin, irinotecan hydrochloride, lanreotide acetate, letrozole, leuprolide acetate, liarozole hydrochloride, lometrexol sodium, lomustine, losoxantrone hydrochloride, masoprocol, maytansine,Mecroteramine hydrochloride, Megestrol acetate, Melenegestrol acetate, Melphalan, Menogaril, Mercaptopurine, Methotrexate, Methotrexate sodium, Metoprine, Meturedepa, Mitindomide, Mitocarcin, Mitocromin, Mitogirin, Mitomalsin, Mitomycin, Mitosper, Mitotan, Mitoxantrone hydrochloride, Mycophenolic acid, Nocodazole, Nogalamycin, Ormaplatin, Oxisuran, Paclitaxel, Pegaspargase, Periomycin, Pentamustine, Pepromycin sulfate, Perfosfamide, Pipobroman, Piposulfan, Pyroxantrone hydrochloride, Plicamycin, Promestane, Porfimer sodium, Porfiromycin prednimustine, Procarbazine hydrochloride, Puromycin, Puromycin hydrochloride, Pyrazofurin, Riboprine, Logretimide, Safingol, Safingol hydrochloride, Semustine, Simtrazene, Sparfosate sodium, Sparsomycin, Spirogermanium hydrochloride, Spirothromycin, Spiroplatin, Streptozocin, Streptozocin, Slofenur, Talisomycin, Tegogalan sodium, Tegafur, Teloxantrone hydrochloride, Temoporfin, Teniposide, Teloxiron, Testolactone, Thiamiprine, Thioguanine, Thiotepa, Thiazofurin, Tiraparazamine, Toremifene citrate, Trestolone acetate, Trisiribine phosphate, Trimethoprim, Trimethoprim glucuronide, Triptorelin, Tubulozole hydrochloride, Uracil mustard, Uredepa, Bapreotide, Verteporfin, Vinblastine sulfate, Vincristine sulfate, Vindesine, Vindesine sulfate, Vinepidine sulfate, Vinglycinate sulfate, Vinroirocin sulfate, Vinorelbine tartrate, Vinrosidine sulfate, Vinzolidine sulfate, Borozole, Zeniplatin, Dinostatin, Zorubicin hydrochloride, but not limited to these. Other anticancer drugs include 20-epi-1,25-dihydroxyvitamin D3, 5-ethynyluracil, Abiraterone, Aclarubicin, Acylfulvene, Adesipenol, Adzelesin, Aldesleukin, ALL-TK antagonist, Altretamine, Ambamustine, Amidox, Amifostine, Aminolevulinic acid, Amrubicin, Amsacrine, Anagrelide,Anastrozole, Andrographolide, Angiogenesis inhibitor, Antagonist D, Antagonist G, Antarelix, Anti-dorsalizing morphogenetic protein 1, Anti-androgen drug, Prostate cancer, Anti-estrogen drug, Antineoplaston, Antisense oligonucleotide, Aphidicolin glycinate, Apoptosis gene regulator, Apoptosis regulator, Applinic acid, ara-CDP-DL-PTBA, Arginine deaminase, Asulacrine, Atamestane, Attrimustine, Axinasatatin 1, Axinasatatin 2, Axinasatatin 3, Azacetron, Azatoxin, Azatyrosine, Baccatin III derivative, Balanol, Batimastat, BCR / ABL antagonist, Benzochlorine, Benzoyl staurosporine, Beta-lactam derivative, Beta-aretin, Betaclamycin B, Betulinic acid, bFGF inhibitor, Bicalutamide, Bisantrene, Bisaziridinyl spermine, Bisnafide, Bistratene A, Bizelesin, Breflate, Broxuridine, Budotitane, Buthionine sulfoximine, Calcipotriol, Calphostin C, Camptothecin derivative, Canarypox IL-2, Capecitabine, Carboxamide-amino-triazole, Carboxamide triazole, CaRestM3, CARN700, Cartilage-derived inhibitor, Carzelesin, Casein kinase inhibitor (ICOS), Castanospermine, Cecropin B, Cetrorelix, Chlorine, Chlorquinoxaline sulfonamide, Cicaprost, Cis-porphyrin, Cladribine, Clomiphene analog, Clotrimazole, Colismycin A, Colismycin B, Combretastatin A4, Combretastatin analog, Conagenin, Crambescidin 816, Crisnatol, Cryptophycin 8, Cryptophycin A derivative, Clacin A, Cyclopentanequinone, Cycloplatam, Sipelamycin, Cytarabine ocfosfate, Cytolytic factor, Cytostatic, Daclizumab, Decitabine, Dehydrodidemnin B, Deslorelin, Dexamethasone, Dexifosfamide, Dexrazoxane, Dexverapamil, Diazicoumarin, Didemnin B, Dodox, Diethylnorspermine, Dihydro-5-azacytidine, Dihydrotaxol, 9-, Dioxamycin, Diphenylspirostaurosporine, Docetaxel, Docosanol, Dolasetron,Doxifluridine, Droloxifene, Dronabinol, Duocarmycin SA, Ebselen, Ecromycin, Edelfosine, Edrecolomab, Efrotomycin, Element, Emitefur, Epirubicin, Epristeride, Estramustine analog, Estrogen agonist, Estrogen antagonist, Ethanidazole, Etoposide phosphate, Exemestane, Fadrozole, Fludarabine phosphate, Fentretinide, Filgrastim, Finasteride, Flavopiridol, Flesinoxan, Fluasterone, Fludarabine, Fluorodoxorubicin hydrochloride, Formestane, Forodesine, Fotemustine, Gadolinium texaphyrin, Gallium nitrate, Galocitabine, Ganirelix, Gelatinase inhibitor, Gemcitabine, Glutathione inhibitor, Hepesulfam, Heregulin, Hexamethylenebisacetamide, Hypericin, Ibandronic acid, Idarubicin, Idoxifene, Idramantane, Ilmofosine, Irinotecan, Imidazoacridone, Imiquimod, Immunostimulatory peptide, Insulin-like growth factor 1 receptor inhibitor, Interferon agonist, Interferon, Interleukin, Yobengamine, Iododoxorubicin, Ipomoeanol, 4-, Iproplatin, Ilsogladine, Isobenzazole, Isohomohalicondrin B, Itasetron, Jasplakinolide, Kahalalide F, Laminarin-N triacetate, Lanreotide, Lomaiviticin, Lenograstim, Lentianan sulfate, Leptostatin, Letrozole, Leukemia inhibitory factor, Leukocyte alpha interferon, Leuprorelin + Estrogen + Progesterone, Leuprorelin, Levamisole, Rialoxifene, Linear polyamine analog, Lipophilic disaccharide peptide, Lipophilic platinum compound, Lissoclinamide 7, Lobaplatin, Lumbricin, Lomustine, Lonidamine, Losoxantrone, Lovastatin, Losoxoribine, Lurtotecan, Lutetium texaphyrin, Lysophospholipin, Soluble peptide, Myotanshinone, Mannostatin A, Marimastat, Masoprocol, Maspin, Matrix metalloproteinase inhibitor, Matrilysin inhibitor, Menogaril, Melvalonate, Meteclorelin, Methioninase, Metoclopramide, MIF inhibitor, Mifepristone, Miltefosine, Miriplostim,Mismatch double-stranded RNA, mitoguazone, mitolactol, mitomycin analogs, mitonafide, mitotoxin fibroblast growth factor-saporin, mitoxantrone, mofarotene, molgramostim, monoclonal antibodies, human chorionic gonadotropin, monophosphoryl lipid A + Mycobacterium cell wall streptokinase, mopidamol, multidrug resistance gene inhibitors, treatment based on multiple tumor suppressor 1, mustard anticancer agents, mycoperooxide B, Mycobacterium cell wall extracts, myriaporon, N-acetyl dinarine, N-substituted benzamides, naphthalene, nagrestip, naloxone + pentazocine, napabucasin, naphterpin, nartograstim, nedaplatin, nemorubicin, neridronic acid, neutral endopeptidase, nilutamide, nisamycin, nitric oxide regulators, nitric oxide antioxidants, nitrulin, O6-benzylguanine, octreotide, oxenone, oligonucleotides, onapristone, ondansetron, ondansetron, oracin, oral cytokine-inducing factors, ormaplatin, osaterone, oxaliplatin, oxaunomycin, paclitaxel, paclitaxel analogs, paclitaxel derivatives, paraureamine, palmitoyl lysophosphatidic acid, pamidronic acid, panaxytriol, panomifene, parabactin, pazelliptine, pegaspargase, perdicine, pentosan polysulfate sodium, pentostatin, pentrozole, perflubron, perfosfamide, perillyl alcohol, phenazinomycin, phenyl acetate, phosphatase inhibitors, picibanil, pilocarpine hydrochloride, pirarubicin, pirimethamine, placetin A, placetin B, plasminogen activator inhibitors, platinum complexes, platinum compounds, platinum-triamine complexes, porfimer sodium, porfiromycin, prednisone, propylbis-acridone, prostaglandin J2, proteasome inhibitors, protein A-based immunomodulators, protein kinase C inhibitors, protein kinase C inhibitors, microalgae, protein tyrosine phosphatase inhibitors, purine nucleoside phosphorylase inhibitors, purpurin, pyrazoloacridine, pyridoxylated hemoglobin polyoxyethylene conjugate, raf antagonists, raltitrexed, ramoseron, ras farnesyl protein transferase inhibitors,ras inhibitor, ras-GAP inhibitor, demethylated retinoic acid, rhenium Re 186 etidronate, lysokinase, ribozyme, RII retinamide, logretimide, rohitukine, romurtide, roquinimex, rubiginone B1, ruboxyl, safingol, sintopine, SarCNU, sarcophytol, ru A, sargramostim, Sdi 1 mimetic, semustine, aging-derived inhibitory factor 1, sense oligonucleotide, signal transduction inhibitor, signal transduction regulator, single-chain antigen-binding protein, sizofiran, sobuzoxane, sodium borocaptate, sodium phenylacetate, sorberol, somatomedin-binding protein, sonermin, sparfosic acid, spicamycin D, spiromustin, sprionpentine, spongistatin 1, squalamine, stem cell inhibitor, stem cell division inhibitor, stipiamide, stromelysin inhibitor, sulfinosine, superactive vasoactive intestinal peptide antagonist, suradista, suramin, swine sonin, synthetic glycosaminoglycan, talimustine, tamoxifen methiodide, tauromustine, tazarotene, tecogalan sodium, tegafur, terlapirillium, telomerase inhibitor, temoporfin, temozolomide, teniposide, tetrachlorodecaoxide, tetrazomine, salibrastine, thiocolchicine, thrombopoietin, thrombopoietin mimetic agent, timalphasin, thrombopoietin receptor agonist, timotrinan, thyroid-stimulating hormone, ethyl etiopurpurin tin, tirapazamine, titanocene dichloride, topsecentin, toremifene, totipotent stem cell factor, translation inhibitor, tretinoin, triacetyluridine, triciribine, trimethoprate, tryptoreline, tropisetron, turosteride, tyrosine kinase inhibitor, tilostatin, UBC inhibitor, ubenimex, urogenital sinus-derived growth inhibitor, urokinase receptor antagonist, vapreotide, variolin B, vector system, erythrocyte gene therapy, veraresol, veramine, bergin, verteporfin, vinorelbine, vincaltine, vitaxin, borozole, zanolteron, zeniplatin, dillascolb, and dinostatin stimalamer, or any combination thereof, but not limited thereto.

[0212] In some embodiments, the anti-cancer agent can be in the prodrug form of the anti-cancer agent. In certain embodiments, the anti-cancer agent can be chemically modified with an alkyl group or an acyl group, or some form of lipid.

[0213] For example, in some embodiments, the toxic injury is a platinum-based anti-tumor agent, a vinca alkaloid agent, an epothilone agent, a taxane agent, a proteasome inhibitor, an immunomodulatory agent, a taxane, cisplatin, radiotherapy for cancer, exposure to radiation at a damage level along the electromagnetic spectrum, a nuclear accident, and a nuclear war, an environmental toxin, or any combination thereof.

[0214] In one embodiment, the toxic injury is a hydrophobic agent. In one embodiment, the toxic injury is a hydrophilic agent. Examples of such toxic injuries include one or more drugs, antibiotics, small molecules, anti-cancer agents, chemotherapeutic agents, immunomodulatory agents, gene silencing agents, medical imaging agents, therapeutic moieties, poorly water-soluble drugs, anti-cancer agents, antibiotics, analgesics, anti-convulsants, anti-diabetic drugs, anti-fungal drugs, anti-tumor drugs, anti-Parkinson's drugs, anti-rheumatic drugs, biological response modifiers, cardiovascular drugs, contrast agents, diagnostic agents, gastrointestinal drugs, ophthalmic drugs, osteoporosis drugs, psychotherapeutic drugs, parasympathomimetics, parasympatholytics, respiratory drugs, sedative-hypnotics, dermatomucosal drugs, smoking cessation drugs, sympatholytics, urinary tract drugs, uterine relaxants, vaginal drugs, vasodilators, antihypertensive drugs, hyperthyroid drugs, anti-hyperthyroid drugs, anti-asthmatic drugs, and anti-dizziness drugs, or any combination thereof, but are not limited thereto.

[0215] In some embodiments, the toxic substance may be an industrial chemical. Toxic industrial chemicals are found in a variety of situations, including manufacturing, agricultural applications, packaging, food additives, and many others. The rate at which new chemicals are produced daily and the importance of developing treatments for the toxicity of various chemical structures underscore the importance of developing treatment strategies to mitigate such toxicity. Since 4-AP has the unexpected efficacy of preventing the toxicity of both paclitaxel and cisplatin (which have very different chemical structures, where cisplatin is chemically similar to industrial chemicals and is similar to taxanes, which are derivatives of industrially produced formulations and chemicals found in nature), and can prevent toxicity to the peripheral nervous system, central nervous system, kidneys, and mitochondria, it is suggested that 4-AP is likely to be useful in a variety of situations beyond those defined in the experimental examples provided by the present invention.

[0216] In some embodiments, the composition further comprises one or more therapeutic agents. In some embodiments, the therapeutic agent is any potassium channel blocker known in the art. Examples of such potassium channel blockers i include, but are not limited to, bretylium, crofilium, dalfampridine, dofetilide, E-4031, ebastine, glibencamide, ibutilide, nifekalant, sematilide, sotalol, sulfonylurea, tedisamil, or any combination thereof.

[0217] In some embodiments, the composition further comprises one antiepileptic drug. Such antiepileptic drugs include, but are not limited to, barbiturates, benzodiazepines, bromides, carbamates, carboxamides, fatty acids, fructose or its derivatives, gamma-aminobutyric acid (GABA) or its analogs, hydantoins, oxazolidinediones, propionates, pyrimidinediones, pyrrolidines, succinimides, sulfonamides, triazines, ureas, valproylamide, or any combination thereof.

[0218] Combination In one embodiment, the composition of the present invention comprises a combination of the agents described herein. In certain embodiments, a composition comprising a combination of the agents described herein has an additive effect, and the overall effect of the combination is approximately equal to the sum of the effects of each individual agent. In other embodiments, a composition comprising a combination of the agents described herein has a synergistic effect, and the overall effect of the combination is greater than the sum of the effects of each individual agent.

[0219] Compositions comprising a combination of agents include the individual agents in any suitable ratio. For example, in one embodiment, the composition comprises two individual agents in a ratio of 1:1. However, the combination is not limited to a particular ratio. Rather, any ratio that has been shown to be effective is encompassed.

[0220] Method of Use In one aspect, the present invention provides a method for preventing, alleviating, and / or treating tissue damage caused by toxic injury, tissue dysfunction caused by toxic injury, mitochondrial dysfunction caused by toxic injury, muscle atrophy caused by toxic injury, neuropathy caused by toxic injury, nephropathy caused by toxic injury, or any combination thereof.

[0221] In one aspect, the present invention provides a method for reducing or restoring tissue damage caused by toxic injury, oxidative damage caused by toxic injury, and / or scarring caused by toxic injury.

[0222] In some embodiments, the tissue damage is multi-tissue damage, multi-organ tissue damage, or any combination thereof. In some embodiments, the tissue damage is kidney tissue damage, liver tissue damage, heart tissue damage, lung tissue damage, brain tissue damage, central nervous system damage, peripheral nerve tissue damage, peripheral neuropathy, nephropathy, chemotherapy-induced peripheral neuropathy (CIPN), radiation-induced peripheral neuropathy (RIPN), chemotherapy-induced nephrotoxicity (CINT), gastrointestinal tract tissue damage, intestinal tissue damage, visual system tissue damage, auditory system tissue damage, skin tissue damage, bladder tissue damage, reproductive system tissue damage, hematopoietic system tissue damage, or any combination thereof. For example, in some embodiments, CIPN is CIPN caused by taxane treatment (P-CIPN), CIPN caused by cisplatin treatment (CisIPN), or any combination thereof.

[0223] In some embodiments, the tissue dysfunction is motor dysfunction, sensory dysfunction, cognitive dysfunction, visual dysfunction, auditory dysfunction, renal dysfunction, hematopoietic system dysfunction, normal skin function, salivary gland dysfunction, liver dysfunction, gallbladder dysfunction, gastrointestinal (GI) dysfunction, sexual dysfunction, or any combination thereof.

[0224] In one aspect, the present invention provides a method for restoring, improving, and / or enhancing at least a portion of tissue function affected by toxic injury, myelination affected by toxic injury, tissue regeneration affected by toxic injury, cell survival affected by toxic injury, neurogenesis affected by toxic injury, repair or regeneration of endogenous stem cells affected by toxic injury, repair or regeneration of transplanted stem cells affected by toxic injury, repair or regeneration of progenitor cells affected by toxic injury, or any combination thereof.

[0225] In some embodiments, the tissue function is a motor function, a sensory function, a cognitive function, a visual function, an auditory function, a renal function, a hematopoietic function, a normal skin function, a salivary gland function, a liver function, a gallbladder function, a gastrointestinal (GI) function, a sexual function, or any combination thereof. In some embodiments, the tissue is kidney tissue, liver tissue, heart tissue, lung tissue, brain tissue, central nervous system tissue, peripheral nerve tissue, gastrointestinal tract tissue, intestinal tissue, visual system tissue, auditory system tissue, skin tissue, bladder tissue, reproductive system tissue, hematopoietic tissue, musculoskeletal tissue, or any combination thereof.

[0226] In one aspect, the present invention provides a method for reducing the size of a lesion affected by toxic injury.

[0227] In one aspect, the present invention provides a method for inhibiting at least one potassium channel blocker for the purpose of treating tissue damage caused by exposure to toxic injury(ies).

[0228] In some embodiments, the method comprises administering to a subject an effective amount of a composition comprising at least one compound (e.g., at least one compound of formula (I)) or a composition of the present invention.

[0229] In one aspect, the present invention provides a method comprising administering to a subject exposed to and / or affected by at least one toxic injury at least one compound or a composition thereof of the present invention. The present invention also provides a method comprising administering to a subject having cancer at least one compound or a composition thereof of the present invention.

[0230] In some embodiments, the present invention provides a method of administering to a subject an effective amount of any of the compounds or pharmaceutical compositions disclosed herein. Accordingly, in some embodiments, the present invention also provides a method comprising administering to a subject exposed to and / or affected by at least one toxic insult an effective amount of any of the compounds or pharmaceutical compositions disclosed herein. In some embodiments, the present invention also provides a method comprising administering to a subject having cancer an effective amount of any of the compounds or pharmaceutical compositions disclosed herein.

[0231] In various embodiments, the toxic insult is any toxic insult described herein. For example, in some embodiments, the toxic insult is a platinum-based anti-tumor agent, a vinca alkaloid agent, an epothilone agent, a taxane agent, a proteasome inhibitor, and an immunomodulatory agent, a taxane, cisplatin, radiation cancer therapy, exposure to radiation at damaging levels along the electromagnetic spectrum, a nuclear accident, and a nuclear war, an environmental toxin, or any combination thereof.

[0232] In some embodiments, treatment can be initiated during treatment of tissue damage that occurs late, at the onset of the injury, during continued exposure to the toxic insult, or after exposure to the toxic insult has ended.

[0233] The compositions of the present invention may be administered systemically, locally, or a combination thereof, to a patient or subject in need thereof.

[0234] The composition of the present invention may be administered to a patient or subject in need thereof by a variety of methods, such as inhalation (e.g., aerosol inhalation), injection, ingestion, oral administration, transdermal administration, blood transfusion, transplantation, sublingual administration, or implantation. The compositions described herein may be administered to a patient subcutaneously, intradermally, intratumorally, intranodally, intramedullarily, intramuscularly, intrathecally, intravenously (i.v.), or intraperitoneally. In one embodiment, the composition is administered systemically to a subject. In one embodiment, the composition of the present invention is administered to a patient by intravenous injection. In one embodiment, the composition is administered locally to a subject. In one embodiment, the composition of the present invention is administered locally to a patient. Any administration may be a single application or multiple applications of the composition of the present invention. The administration may be to a single site or multiple sites of the individual being treated. Multiple administrations may be carried out essentially simultaneously or at intervals over time.

[0235] Subjects in which administration of the pharmaceutical composition of the present invention is contemplated include, but are not limited to, mammals such as humans and other primates, non-human primates, cattle, pigs, horses, sheep, cats, dogs, etc.

[0236] The pharmaceutical composition of the present invention may be administered by a method suitable for the disease to be treated (or prevented). The dosage and dosing frequency are determined by factors such as the condition of the subject, the type and severity of the disease of the subject, etc., but appropriate dosages may also be determined by clinical trials.

[0237] When a "therapeutic amount" is indicated, the exact amount of the composition of the present invention to be administered can be determined by a physician taking into account individual differences such as the age, weight, type of disease, degree of disease, and condition of the patient (subject).

[0238] Formulation / Pharmaceutical Composition The present invention also encompasses the use of a pharmaceutical composition comprising a compound of the present invention or a composition thereof. Such a pharmaceutical composition may comprise at least one compound of the present invention in a form suitable for administration to a subject, or the pharmaceutical composition may comprise at least one compound of the present invention and one or more pharmaceutically acceptable carriers, one or more additional ingredients, or combinations thereof. The compounds of the present invention may be present in the pharmaceutical composition in the form of physiologically acceptable salts, such as in combination with physiologically acceptable cations or anions, as is well known in the art.

[0239] Administration of a therapeutic agent according to the present invention may be continuous or intermittent, depending, for example, on the physiological state of the recipient, whether the purpose of administration is therapeutic or prophylactic, and other factors known to those of skill in the art. Administration of the agents of the present invention may be essentially continuous over a preselected period of time or may be a series of spaced doses. Both local and systemic administration are contemplated. The dosage will vary depending upon the selected composition, the particular disease, body weight, physical condition, and age of the subject, and whether prophylaxis or therapy is to be achieved, among other factors. Such factors can be readily determined by a clinician using animal models or other test systems well known in the art.

[0240] Administration of the composition of the present invention in a method of treatment can be accomplished in a variety of ways using methods known in the art. In one embodiment, the method of the present invention comprises systemic administration to a subject, including, for example, enteral or parenteral administration. In a particular embodiment, the method comprises intradermal delivery of the composition. In another embodiment, the method comprises intravenous delivery of the composition. In some embodiments, the method comprises intramuscular delivery of the composition. In one embodiment, the method comprises subcutaneous delivery of the composition. In one embodiment, the method comprises inhalation of the composition. In one embodiment, the method comprises intranasal delivery of the composition.

[0241] Depending on the intended mode of administration, the pharmaceutical composition is preferably in unit dosage form suitable for single administration of an exact dose, for example, in the form of solids, semi-solids, liquids, solutions, suspensions (e.g., those incorporated into microparticles, liposomes, etc.), emulsions, gels, etc. The pharmaceutical composition contains, as described above, an effective amount of a potassium channel blocker such as a 4-aminopyridine compound, its derivatives, or combinations thereof, in combination with a pharmaceutically acceptable carrier, and may further contain other carriers, adjuvants, diluents, thickeners, buffers, preservatives, surfactants, etc. The pharmaceutical composition may contain one or more additional active ingredients such as other drugs, pharmaceuticals, antibacterial agents, anti-inflammatory agents, anesthetics, antiepileptic agents, etc.

[0242] When appropriate, the formulation is conveniently provided in individual unit dosage form and can be prepared by any method well known in pharmacy. Such methods can include associating the therapeutic agent with a liquid carrier, solid matrix, semi-solid carrier, finely divided solid carrier, or combinations thereof, and then, if necessary, introducing or shaping the product into the desired delivery system.

[0243] Accordingly, the therapeutic and prophylactic methods of the present invention include the use of a pharmaceutical composition containing at least one compound of the present invention for carrying out the method of the present invention. The pharmaceutical composition useful for practicing the present invention may be administered to deliver a dose of from 0.001 ng / kg / day to 100 mg / kg / day. For example, in some embodiments, the pharmaceutical composition useful for practicing the present invention may be administered to deliver a dose of from 0.005 mg / kg / day to 5 mg / kg / day. In one embodiment, the present invention includes administration at a dose such that the concentration of the compound of the present invention in the serum of a mammal is from 10 nM to 10 μM.

[0244] Generally, the dosage that may be administered to a mammal, preferably a human, in the method of the present invention ranges from 0.01 μg to about 50 mg per kilogram of the mammal's body weight. However, the exact dosage to be administered varies depending on a number of factors including, but not limited to, the type of mammal and the type of disease state being treated, the age of the mammal, the route of administration, and the specific agent(s) being used. Preferably, the dosage of the compound varies from about 0.1 μg to about 10 mg per kilogram of the mammal's body weight. More preferably, the dosage varies from about 1 μg to about 5 mg per kilogram of the mammal's body weight. For example, in some embodiments, the dosage varies from about 0.005 mg to about 5 mg per kilogram of the mammal's body weight.

[0245] The relative amounts of the active ingredient, pharmaceutically acceptable carrier, and any additional ingredients in the pharmaceutical composition of the present invention vary depending on the identity, size, and condition of the subject being treated and, further, on the route by which the composition is to be administered. By way of example, the composition can contain from 0.1% to 100% (w / w) of the active ingredient.

[0246] The pharmaceutical composition can contain from about 0.01 to about 99 percent of a potassium channel blocker (e.g., 4-aminopyridine or a derivative or analog thereof) together with a carrier and / or excipient. For example, the amount of the potassium channel blocker (e.g., 4-aminopyridine, a derivative thereof, or a combination thereof) can be about 0.1% or more, about 1% or more, about 2% or more, about 5% or more, about 10% or more, about 15% or more, about 20% or more, about 25% or more, about 30% or more, about 50% or more, about 75% or more, or about 90% or more by weight of the pharmaceutical composition.

[0247] In some embodiments, the pharmaceutical composition is defined by its ability to achieve a therapeutically effective serum concentration of 4-aminopyridine or a derivative thereof. In some embodiments, such a concentration ranges from about 10 nM to about 1 μM of 4-aminopyridine, or a higher concentration when combined with an anti-seizure agent. The desired serum concentration of a 4-aminopyridine derivative is defined by the ability of such an agent to provide the desired therapeutic effect without causing unacceptable side effects.

[0248] The pharmaceutical compositions described herein are used with a "therapeutically effective amount" of a potassium channel blocker (e.g., 4-aminopyridine, a derivative thereof, or a combination thereof). In some embodiments, the pharmaceutical composition can be formulated to deliver a therapeutically effective amount of a potassium channel blocker (e.g., 4-aminopyridine, a derivative thereof, or a combination thereof) in an amount of 2.5 mg or more upon administration. For example, upon administration, the pharmaceutical composition can deliver 3 mg or more, 4 mg or more, 5 mg or more, 6 mg or more, 7 mg or more, 7.5 mg or more, 8 mg or more, 9 mg or more, 10 mg or more, 15 mg or more, 20 mg or more, 25 mg or more, 30 mg or more, 35 mg or more, 40 mg or more, 45 mg or more, 50 mg or more, 55 mg or more, 60 mg or more, 65 mg or more, 70 mg or more, 75 mg or more, 80 mg or more, 85 mg or more, 90 mg or more, or 95 mg or more of a potassium channel blocker (e.g., 4-aminopyridine, a derivative thereof, or a combination thereof).

[0249] The composition may be administered to a mammal several times daily, or at a lower frequency, such as once a day, once a week, once every two weeks, once a month, or even at a lower frequency, such as once every few months or even less than once a year. The dosing frequency will be readily apparent to those skilled in the art and depends on many factors, including but not limited to the type and severity of the disease being treated, the type and age of the mammal, whether the administration is used for the treatment of a dysfunction or for the identification of individuals for whom a complete course of treatment would be beneficial.

[0250] In another aspect of the present invention, using the methods disclosed herein, individuals who would benefit from treatment with a potassium channel blocker (e.g., 4-AP, a derivative of 4-AP, or any combination thereof) can be identified. In some embodiments, an individual who exhibits dysfunction in one or more tissues after exposure to an artificially generated toxic insult may be treated for 1 to 10 days using the methods disclosed herein, and it may be determined whether the tissue function is improved by the treatment. Thus, in some embodiments, the methods disclosed herein can be used to provide individualized and targeted therapy.

[0251] When preparing the therapeutic agent of the present invention for administration, preferably, it is combined with a pharmaceutically acceptable carrier, diluent, or excipient to form a pharmaceutical preparation or unit dosage form. The total active ingredient in such a preparation occupies 0.1 to 99.9% of the weight of the preparation. A "pharmaceutically acceptable" carrier, diluent, or excipient is a carrier, diluent, excipient, and / or salt that is compatible with the other components of the preparation and is not harmful to its recipient. The active ingredient for administration may be present in the form of a powder or granule, solution, suspension, or emulsion.

[0252] The pharmaceutical preparation containing the therapeutic agent of the present invention can be prepared by procedures known in the art using well-known and readily available ingredients. The therapeutic agent of the present invention can also be formulated, for example, as a solution suitable for parenteral administration by intramuscular, subcutaneous, or intravenous routes.

[0253] The pharmaceutical preparation of the therapeutic agent of the present invention can also take the form of an aqueous or anhydrous solution or dispersion, or alternatively, an emulsion or suspension.

[0254] Accordingly, the therapeutic agent may be formulated for parenteral administration (e.g., by injection, e.g., by bolus injection or continuous infusion), and may be provided in unit dosage forms in ampoules, prefilled syringes, small volume infusion containers, or multi-dose containers with added preservatives. The formulation of a pharmaceutical composition suitable for parenteral administration is a combination of an active ingredient and a pharmaceutically acceptable carrier such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or sold in a form suitable for bolus or continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage forms such as ampoules and multi-dose containers containing preservatives. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained release or biodegradable formulations. Such formulations may further contain one or more additional components including, but not limited to, suspending agents, stabilizers, or dispersing agents. In one embodiment of a formulation for parenteral administration, the active ingredient is provided in a dry (i.e., powder or granule) form for reconstitution with a suitable vehicle (e.g., sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition.

[0255] It will be understood that the unit content of the active ingredient contained in each aerosol dose of each dosage form need not itself constitute an effective amount for treating a particular indication or disease, since the required effective amount can be achieved by administering a plurality of dosage units. Further, the effective amount may be achieved by using an amount less than the dosage in the dosage form, either individually or in a series of administrations.

[0256] The pharmaceutical formulations of the present invention may include, as optional components, pharmaceutically acceptable carriers, diluents, solubilizers or emulsifiers, and salts of the types well known in the art. Specific non-limiting examples of carriers and / or diluents useful in the pharmaceutical formulations of the present invention include water and physiologically acceptable buffered saline, such as phosphate buffered saline at pH 7.0 to 8.0.

[0257] The compounds of the present invention can be formulated and administered for treating various disease states by any means that result in contact between the site of action of the agent in the organism and the active ingredient. These can be administered by any conventional means available for use in combination with pharmaceuticals, either as individual therapeutic active ingredients or as combinations of therapeutic active ingredients. They can be administered alone, but are usually administered together with a pharmaceutical carrier selected based on the chosen route of administration and standard pharmaceutical practice.

[0258] Generally, water, suitable oils, physiological saline, aqueous dextrose (glucose) solutions, and related sugar solutions, as well as glycols such as propylene glycol or polyethylene glycol, are carriers suitable for parenteral solutions. Solutions for parenteral administration contain the active ingredient, suitable stabilizers, and, if necessary, buffering substances. Antioxidants such as sodium bisulfate, sodium sulfite, or ascorbic acid are used as suitable stabilizers, either alone or in combination. Citric acid and its salts, and sodium ethylenediaminetetraacetate (EDTA) are also used. Additionally, parenteral solutions can contain preservatives such as benzalkonium chloride, methylparaben or propylparaben, and chlorobutanol. Suitable pharmaceutical carriers are described in Remington’s Pharmaceutical Sciences, a standard reference in the field.

[0259] Furthermore, the duration of action can be controlled using standard pharmaceutical methods. These are well known in the art and include controlled release formulations and may include suitable polymers such as polymers, polyesters, polyamino acids, polyvinyls, pyrrolidone, ethylene vinyl acetate, methyl cellulose, carboxymethyl cellulose, or protamine sulfate. To control the release, the concentration of the polymer as well as the incorporation method can be adjusted. Furthermore, the drug can be incorporated into particles of polymer materials such as polyesters, polyamino acids, hydrogels, poly(lactic acid), or ethylene vinyl acetate copolymers. In addition to incorporation, these drugs can also be used to trap compounds in microcapsules.

[0260] Accordingly, the pharmaceutical compositions of the present invention can be delivered to various sites within the body of a mammal via various routes to achieve specific effects (see, for example, Rosenfeld et al., 1991; Rosenfeld et al., 1991a; Jaffe et al., supra; Berkner, supra). One of ordinary skill in the art will recognize that multiple routes can be used for administration, but that a particular route may provide a more immediate and effective response than another. Local or systemic delivery can be achieved by administration including application or instillation of the formulation into a body cavity, inhalation or insufflation of an aerosol, or parenteral introduction including intramuscular, intravenous, peritoneal, subcutaneous, intradermal, and topical administration.

[0261] The active ingredient of the present invention can be provided in unit dosage forms, and each dosage unit, for example, a teaspoonful, tablet, solution, or suppository, contains a predetermined amount of the composition, either alone or in suitable combination with other active ingredients. As used herein, the term "unit dosage form" refers to physically discrete units suitable as unit doses for human and mammalian subjects, each unit containing, when appropriate, a pharmaceutically acceptable diluent, carrier, or vehicle, and a predetermined amount of the composition of the present invention calculated to produce the desired effect, alone or in combination with other active agents. The specifications of the unit dosage forms of the present invention depend on the particular effect to be achieved and the particular pharmacodynamics associated with the pharmaceutical composition in a particular host.

[0262] In one embodiment, the composition of the present invention is formulated using one or more pharmaceutically acceptable excipients or carriers. In one embodiment, the pharmaceutical composition of the present invention comprises a therapeutically effective amount of a compound or complex of the present invention and a pharmaceutically acceptable carrier. Useful pharmaceutically acceptable carriers include, but are not limited to, glycerol, water, saline, ethanol, and other pharmaceutically acceptable salt solutions, such as salts of phosphates and organic acids. Examples of these and other pharmaceutically acceptable carriers are described in Remington’s Pharmaceutical Sciences (1991, Mack Publication Co., New Jersey).

[0263] The carrier may be, for example, a solvent or dispersion medium containing water, ethanol, polyols (such as glycerol, propylene glycol, and liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. Appropriate fluidity can be maintained, for example, by using coatings such as lecithin, by maintaining the particle size required in the case of a dispersion, and by using surfactants. Prevention of microbial activity can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc. In many cases, it is preferable to include in the composition an isotonic agent, such as saccharides, sodium chloride, or polyhydric alcohols such as mannitol and sorbitol. Sustained absorption of the injectable composition can be brought about by including in the composition an agent that delays absorption, such as aluminum monostearate or gelatin. In one embodiment, the pharmaceutically acceptable carrier is not DMSO alone.

[0264] The present invention also provides a pharmaceutical composition comprising one or more of the compositions described herein. The formulation may be used in admixture with conventional excipients, i.e., pharmaceutically acceptable organic or inorganic carrier substances suitable for administration to the subject. The pharmaceutical composition can be sterilized and, if desired, mixed with adjuvants such as lubricants, preservatives, stabilizers, wetting agents, emulsifying agents, salts for influencing the osmotic buffer, coloring agents, and / or aromatic substances, etc. They may be combined, if desired, with other active agents, such as analgesics or anticonvulsants.

[0265] As used herein, "additional components" include, but are not limited to, one or more of the following: excipients, surfactants, dispersants, inert diluents, granulating agents and disintegrants, binders, lubricants, colorants, preservatives, physiologically degradable compositions such as gelatin, aqueous vehicles and solvents, oily vehicles and solvents, suspending agents, dispersing or wetting agents, emulsifying agents, viscous agents, buffering agents, salts, thickening agents, fillers, emulsifying agents, antioxidants, antibiotics, antifungal agents, stabilizers, and pharmaceutically acceptable polymeric or hydrophobic materials. Other "additional components" that may be included in the pharmaceutical compositions of the present invention are known in the art and are described, for example, in Genaro, ed. (1985, Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, PA), which is hereby incorporated by reference.

[0266] The compositions of the present invention may contain a preservative in an amount of from about 0.005% to 2.0% by weight, based on the total weight of the composition. Preservatives are used to prevent spoilage when the composition is exposed to contaminants in the environment. Examples of useful preservatives according to the present invention include, but are not limited to, those selected from benzyl alcohol, sorbic acid, parabens, imidurea, or any combination thereof. Particularly preferred preservatives are a combination of about 0.5% to 2.0% benzyl alcohol and 0.05% to 0.5% sorbic acid.

[0267] In one embodiment, the composition comprises an antioxidant and a chelating agent that inhibits the decomposition of one or more components of the composition. Preferred antioxidants for some compounds are BHT, BHA, α-tocopherol, and ascorbic acid in a preferred range of about 0.01% to 0.3% based on the total weight of the composition, and more preferably BHT in the range of 0.03% by weight to 0.1% by weight. Preferably, the chelating agent is present at 0.01% to 0.5% by weight based on the total weight of the composition. Particularly preferred chelating agents include edetates (e.g., disodium edetate) and citric acid in the range of about 0.01% to 0.20% by weight, more preferably in the range of 0.02% to 0.10% by weight, based on the total weight of the composition. Chelating agents are useful for chelating metal ions in the composition that can be detrimental to the shelf life of the formulation. BHT and disodium edetate are particularly preferred antioxidants and chelating agents, respectively, for some compounds, but other suitable and equivalent antioxidants and chelating agents may be substituted as known to those skilled in the art.

[0268] The pharmaceutical compositions useful in the methods of the present invention may be prepared, packaged, or sold in formulations suitable for administration by eye, oral, rectal, vaginal, parenteral, topical, pulmonary, intranasal, buccal, intravenous, intraventricular, intradermal, intramuscular, or another route of administration. Other contemplated formulations include intravaginal pessaries, liposome preparations, resealed erythrocytes containing the active ingredient, and immunogenic-based formulations.

[0269] The pharmaceutical compositions of the present invention may be prepared, packaged, or sold in bulk, as a single unit dose, or as multiple unit doses. As used herein, a "unit dose" is an individual quantity of a pharmaceutical composition containing a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient to be administered to a subject, or a convenient fraction of such a dose, e.g., one-half or one-third of such a dose.

[0270] The formulations of the pharmaceutical compositions described herein can be prepared by any method known in the field of pharmacology or developed in the future. Generally, such preparation methods include the step of associating the active ingredient with a carrier or one or more other auxiliary components, and then, if necessary or desirable, shaping or packaging the product into the desired single or multiple dosage units.

[0271] In an exemplary embodiment, the pharmaceutical composition includes a pharmaceutically acceptable excipient such as a pharmaceutically acceptable carrier, and an exemplary compound described herein.

[0272] In certain exemplary embodiments, the pharmaceutical composition generally includes a pharmaceutically acceptable salt as described below or is in the form of such a salt.

[0273] Exemplary compounds can be administered in the form of prodrugs. A prodrug can include a covalently bonded carrier that releases the active parent drug when administered to a mammalian subject. Prodrugs can be prepared by modifying the functional groups present in the compound such that routine manipulations or in vivo modifications are cleaved to yield the parent compound. Prodrugs include, for example, compounds in which a hydroxyl group is bonded to any group that, when administered to the subject, is cleaved to form a free hydroxyl group.

[0274] The description of the pharmaceutical compositions provided herein mainly relates to pharmaceutical compositions suitable for ethical administration to humans, but it will be understood by those skilled in the art that such compositions are generally suitable for administration to all kinds of animals. Modifications to make pharmaceutical compositions suitable for administration to humans suitable for administration to various animals are well understood, and an ordinary skilled veterinary pharmacologist can design and perform such modifications with only ordinary experimentation or by performing experiments. The subjects to which administration of the pharmaceutical compositions of the present invention is contemplated include mammals, including but not limited to humans and other primates, non-human primates, cattle, pigs, horses, sheep, cats, and dogs, and other commercially relevant mammals.

[0275] The controlled-release or sustained-release formulations of the pharmaceutical compositions of the present invention may be manufactured using conventional techniques.

[0276] The pharmaceutical composition may be prepared, packaged, or sold in the form of a sterile injectable aqueous or oily suspension or solution. This suspension or solution may be formulated according to known techniques and may contain additional components such as dispersants, wetting agents, or suspending agents described herein in addition to the active ingredient. Such sterile injectable formulations may be prepared using a non-toxic parenterally acceptable diluent or solvent, such as water or 1,3-butanediol. Other acceptable diluents and solvents include, but are not limited to, Ringer's solution, isotonic sodium chloride solution, and fixed oils such as synthetic monoglycerides or diglycerides. Other useful parenteral dosage forms include those containing the active ingredient in microcrystalline form, in liposome preparations, or as a component of a biodegradable polymer system. Sustained-release or implantable compositions may contain pharmaceutically acceptable polymeric or hydrophobic materials, such as emulsions, ion exchange resins, poorly soluble polymers, or poorly soluble salts.

[0277] Formulations of pharmaceutical compositions suitable for parenteral administration are combinations of the active ingredient with a pharmaceutically acceptable carrier such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or sold in a form suitable for bolus administration or for continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage form, such as in ampoules or in multi-dose containers containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable formulations. Such formulations may further contain one or more additional components including, but not limited to, suspending agents, stabilizers, or dispersants. In one embodiment of a formulation for parenteral administration, the active ingredient is provided in dry (i.e., powder or granular) form for reconstitution with a suitable vehicle (e.g., sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition.

[0278] Transdermal formulations can also be prepared in the form of creams, ointments, pastes, sprays, gels, lotions, emulsions, and transdermal patches. Such compositions can contain one or more chemical penetration enhancers, membrane permeability agents, membrane transport agents, emollients, surfactants, stabilizers, and combinations thereof.

[0279] A pharmaceutical composition may be prepared, packaged, or sold in the form of a sterile injectable aqueous or oily suspension or solution. This suspension or solution may be formulated according to known techniques and may contain, in addition to the active ingredient, additional ingredients such as dispersing agents, wetting agents, or suspending agents described herein. Such sterile injectable formulations may be prepared using a non-toxic parenterally acceptable diluent or solvent, such as water or 1,3-butanediol. Other acceptable diluents and solvents include, but are not limited to, Ringer's solution, isotonic sodium chloride solution, and fixed oils such as synthetic monoglycerides or diglycerides. Other useful parenteral dosage forms include those containing the active ingredient in microcrystalline form, in liposome preparations, or as a component of a biodegradable polymer system. Sustained release or implantable compositions may contain pharmaceutically acceptable polymeric or hydrophobic materials, such as emulsions, ion exchange resins, poorly soluble polymers, or poorly soluble salts.

[0280] Liquid pharmaceutically administrable compositions can be prepared, for example, by dissolving, dispersing, etc., a composition described herein and any pharmaceutical adjuvants in excipients such as water, physiological saline, aqueous dextrose solution, glycerol, ethanol, etc. to form a solution or suspension. Optionally, the pharmaceutical composition to be administered may contain trace amounts of non-toxic auxiliary substances such as wetting agents or emulsifying agents, pH buffering agents, etc., such as sodium acetate, sorbitan monolaurate, sodium triethanolamine acetate, sodium triethanolamine oleate, etc. The actual methods for preparing such dosage forms are known or apparent to those skilled in the art; see, for example, "Remington’s Pharmaceutical Sciences" mentioned above.

[0281] The liquid suspension may be prepared by suspending the HMW-HA or other compositions of the present invention in an aqueous or oily vehicle using conventional methods. Examples of aqueous vehicles include water and isotonic saline. Examples of oily vehicles include almond oil, oily esters, ethyl alcohol, vegetable oils such as peanut oil, olive oil, sesame oil, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin. The liquid suspension may further contain one or more additional components including, but not limited to, suspending agents, dispersing or wetting agents, emulsifying agents, lubricants, preservatives, buffers, salts, flavorings, colorants, and sweeteners. Oily suspensions may further contain thickening agents. Known suspending agents include, but are not limited to, sorbitol syrup, hydrogenated edible fats, sodium alginate, polyvinylpyrrolidone, tragacanth gum, acacia gum, and cellulose derivatives such as sodium carboxymethylcellulose, methylcellulose, and hydroxypropylmethylcellulose. Known dispersing or wetting agents include naturally occurring phospholipids such as lecithin, condensation products of alkylene oxides with fatty acids, long-chain aliphatic alcohols, partial esters derived from fatty acids and hexitols, or partial esters derived from fatty acids and hexitol anhydrides (e.g., polyoxyethylene stearate, heptadecaethyleneoxy cetanol, polyoxyethylene sorbitol monooleate, and polyoxyethylene sorbitan monooleate, respectively). Known emulsifying agents include, but are not limited to, lecithin and acacia. Known preservatives include, but are not limited to, methyl parahydroxybenzoate, ethyl parahydroxybenzoate, or n-propyl parahydroxybenzoate, ascorbic acid, and sorbic acid.

[0282] The powder and granule formulations of the pharmaceutical preparations of the present invention can be prepared using known methods. Such formulations can be administered directly to a subject or used, for example, to form tablets, fill capsules, or prepare aqueous or oily suspensions or solutions by adding an aqueous or oily vehicle thereto. Each of these formulations may further contain one or more of a dispersing or wetting agent, a suspending agent, and a preservative. Additional excipients, such as fillers and sweeteners, flavoring agents, or coloring agents, may also be included in these formulations.

[0283] Dry powder formulations (「DPF」) with large particle sizes have improved flowability characteristics, such as less aggregation, easy aerosolization, and potentially less phagocytosis. Dry powder aerosols for inhalation therapy are generally manufactured mainly with an average diameter in the range of less than 5 microns, but the preferred range is an aerodynamic diameter of 1 - 10 microns. Large 「carrier」 particles (drug-free) are co-delivered with the therapeutic aerosol to assist in achieving efficient aerosolization, among other possible advantages.

[0284] The pharmaceutical compositions of the present invention can also be prepared, packaged, or sold in the form of water-in-oil emulsions or oil-in-water emulsions. The oil phase can be a vegetable oil such as olive oil or peanut oil, a mineral oil such as liquid paraffin, or a combination thereof. Such compositions may further contain one or more emulsifying agents, such as natural gums like gum acacia or tragacanth gum, natural phosphatides like soy or lecithin phosphatide, esters or partial esters derived from combinations of fatty acids and hexitol anhydrides such as sorbitan monooleate, and condensation products of such partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. These emulsions may also contain additional ingredients, for example, sweetening or flavoring agents.

[0285] Methods for impregnating or coating a material with a chemical composition are known in the art and include methods of depositing or bonding a chemical composition onto a surface, incorporating a chemical composition into the structure of a material during synthesis (i.e., in a physiologically degradable material), and absorbing an aqueous or oily solution or suspension into an absorbent material, with or without subsequent drying, but are not limited thereto.

[0286] The dosing regimen can affect what constitutes an effective amount. The therapeutic agent may be administered to the subject either before or after diagnosis of the disease. Further, several divided doses, and alternating doses, may be administered daily or continuously, or the dose may be continuously infused, or may be a bolus injection. Further, the dose of the therapeutic agent may be proportionally increased or decreased as indicated by the urgency of the therapeutic or prophylactic situation.

[0287] Preferably, administration of the composition of the present invention to a subject, including a mammal, more preferably a human, may be carried out in an effective dose and for a period effective to prevent or treat a disease using known procedures. The effective amount of the therapeutic compound required to achieve a therapeutic effect will vary depending on factors such as the activity of the particular compound employed, the time of administration, the rate of excretion of the compound, the duration of the treatment, other drugs, compounds, or materials used in combination with the compound, the condition of the disease or disorder of the subject being treated, age, sex, weight, medical condition, general health status, and previous medical history, as well as similar factors well known in the medical arts. The dosing regimen may be adjusted to obtain an optimal therapeutic response. For example, several divided doses may be administered daily for several days, or the dose may be proportionally reduced when the urgency of the treatment situation is indicated. A non-limiting example of the effective dose range of the therapeutic compound of the present invention is about 1 to 5,000 mg / kg of body weight per day. One of ordinary skill in the art will be able to study the relevant factors and make a determination regarding the effective amount of the therapeutic compound without undue experimentation.

[0288] The compound may be administered to the subject several times a day, or at less frequent intervals such as once a day, once a week, once every two weeks, once a month, or even less frequently, such as once every few months, once a year or less. In non-limiting examples, it is understood that the daily dose of the compound may be administered daily, every other day, every two days, every three days, every four days, or every five days. For example, in the case of alternate-day administration, a dose of 5 mg / day may be started on Monday, the first subsequent dose of 5 mg / day may be administered on Wednesday, and the second subsequent dose of 5 mg / day may be administered on Friday, and so on. The dosing frequency will be readily apparent to those skilled in the art and will depend on several factors, such as, but not limited to, the type and severity of the disease being treated, the type and age of the animal, etc.

[0289] The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention can vary to obtain an amount of the active ingredient that is effective in achieving the desired therapeutic response for a particular subject, composition, and mode of administration without being toxic to the subject.

[0290] A physician having ordinary skill in the art, such as a medical doctor or veterinarian, can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian can start with a dose of the compound of the present invention employed in the pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0291] In certain embodiments, it is particularly advantageous to formulate the compounds into unit dosage forms for ease of administration and uniformity of dosage. As used herein, a unit dosage form refers to physically discrete units suitable as unit dosages for a subject to be treated, each unit containing a predetermined amount of a therapeutic compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical vehicle. The unit dosage forms of the present invention are dependent upon and directly determined by (a) the particular characteristics of the therapeutic compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in and peculiar to the art of compounding / formulating such therapeutic compounds for the treatment of diseases in a subject.

[0292] In one embodiment, the compositions of the present invention are administered to a subject at a dosage in the range of from once to more than five times per day.

[0293] In another embodiment, the compositions of the present invention are administered to a subject at dosages in the range including, but not limited to, once daily, once every two days, once every three days to once a week, and once every two weeks. In another embodiment, aiming to maintain a therapeutic serum concentration of a therapeutic agent that is therapeutically effective without inducing unacceptable side effects, depending on the exact sustained-release formulation used, the compositions of the present invention are administered to a subject at dosages in the range applicable to sustained-release formulations including, but not limited to, once every two days, once every three days to once a week, and once every two weeks.

[0294] It will be readily apparent to those skilled in the art that the frequency of administration of the various combination compositions of the present invention will vary for each subject depending on a number of factors including, but not limited to, age, the disease or disorder being treated, sex, general health, and other factors. Accordingly, the present invention should not be construed as being limited to any particular dosing regimen, and the exact dosage and composition to be administered to any subject will be determined by the attending physician taking into account all other factors relevant to the subject.

[0295] The compounds of the present invention for administration can be in the range of about 1 mg to about 10,000 mg, about 20 mg to about 9,500 mg, about 40 mg to about 9,000 mg, about 75 mg to about 8,500 mg, about 150 mg to about 7,500 mg, about 200 mg to about 7,000 mg, about 3050 mg to about 6,000 mg, about 500 mg to about 5,000 mg, about 750 mg to about 4,000 mg, about 1 mg to about 3,000 mg, about 10 mg to about 2,500 mg, about 20 mg to about 2,000 mg, about 25 mg to about 1,500 mg, about 50 mg to about 1,000 mg, about 75 mg to about 900 mg, about 100 mg to about 800 mg, about 250 mg to about 750 mg, about 300 mg to about 600 mg, about 400 mg to about 500 mg, and any whole or partial increments therebetween.

[0296] In some embodiments, the dosage of the compound of the present invention is from about 1 mg to about 2,500 mg. In some embodiments, the dosage of the compound of the present invention used in the compositions described herein is less than about 10,000 mg, or less than about 8,000 mg, or less than about 6,000 mg, or less than about 5,000 mg, or less than about 3,000 mg, or less than about 2,000 mg, or less than about 1,000 mg, or less than about 500 mg, or less than about 200 mg, or less than about 50 mg. Similarly, in some embodiments, the dosage of the second compound (i.e., the drug used to treat the same or another disease as the disease treated by the composition of the present invention) described herein is less than about 1,000 mg, less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 400 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 40 mg, or less than about 30 mg, or less than about 25 mg, or less than about 20 mg, or less than about 15 mg, or less than about 10 mg, or less than about 5 mg, or less than about 2 mg, or less than about 1 mg, or less than about 0.5 mg, and any whole or part of all such increments.

[0297] In one embodiment, the present invention relates to a packaged pharmaceutical composition comprising a container holding a therapeutically effective amount of a compound or complex of the present invention, alone or in combination with a second agent, and instructions for using the compound or complex to treat, prevent, or reduce one or more symptoms of a disease in a subject.

[0298] The term "container" includes any receptacle for containing the pharmaceutical composition. For example, in one embodiment, the container is the packaging containing the pharmaceutical composition. In other embodiments, the container is not the packaging containing the pharmaceutical composition, i.e., the container is a container such as a box or vial that contains the packaged pharmaceutical composition or the unpackaged pharmaceutical composition and instructions for use of the pharmaceutical composition. Further, packaging techniques are well known in the art. It should be understood that the instructions for use of the pharmaceutical composition may be included in the packaging containing the pharmaceutical composition, and thus the instructions form a strong functional relationship with the packaged product. However, it is necessary to understand that the instructions may also include information regarding the ability of the compound to perform its intended function, such as treating or preventing a disease in a subject, or delivering a contrast agent or diagnostic agent to a subject.

[0299] Routes of administration of the compositions of the present invention include oral, nasal, rectal, parenteral, sublingual, transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, vaginal (e.g., transvaginal and perivaginal), (intra)nasal, and (trans)rectal), intravesical, intralung, intraduodenal, intragastric, intrathecal, subcutaneous, intramuscular, intradermal, intraarterial, intravenous, intratracheal, inhalation, and topical administration.

[0300] Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gel capsules, lozenges, dispersants, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magma, pastilles, creams, pastes, plasters, lotions, disks, suppositories, liquid sprays for nasal or oral administration, dry powders or aerosolized formulations for inhalation, compositions and formulations for intravesical administration, and the like. It should be understood that the formulations and compositions that may be useful in the present invention are not limited to the specific formulations and compositions described herein.

[0301] In some examples, the composition can take the form of beads, films, or other shapes, as understood by those skilled in the art. The size of the individual beads, films, or other shapes is a size suitable for implantation or other administration forms, as understood by those skilled in the art. Further, the size of the individual beads, films, or other shapes may be substantially uniform or there may be a distribution of different sizes for each shape. Optionally, the beads are implanted into the body, ingested, or placed in the body in some way to administer the drug locally or systemically in a sustained-release manner.

[0302] It will be understood that the compositions of the present invention may be administered to a subject alone or in combination with another drug.

[0303] In certain embodiments, administration of the compositions of the present invention may be by a single administration or boosted by multiple administrations.

[0304] In one embodiment, the present invention includes a method comprising administering a combination of the compounds described herein. In certain embodiments, the combination has an additive effect and the total effect of administering the combination is approximately equal to the sum of the effects of administering each compound. In other embodiments, the combination has a synergistic effect and the total effect of administering the combination is greater than the sum of the effects of administering each compound.

[0305] Optionally, the composition may include a formulation suitable for delivering the treatment as a sustained release formulation that can release the therapeutic substance over a period of hours to weeks. Such sustained release formulations can consist of osmotic pumps, fibrin adhesives, biocompatible polymers or hydrogels, or other means of delivering the treatment in a formulation that allows for sustained release. The compound can be encapsulated within a polymer or hydrogel, whereby the drug is slowly released in the body to at least a portion of the tissue(s) damaged by the toxic insult. Optionally, the compound can be dispersed throughout the polymer or hydrogel in such a way as to provide a slow sustained release as the polymer or hydrogel degrades in the body. In some embodiments, the composition includes biodegradable biocompatible polymers such as polyglycolide or polyglycolic acid (PGA), polylactide or polylactic acid (PLA), poly-L-lactic acid (PLLA), poly-D / L-lactic acid and polyglycolic acid (PDLLA-co-PGA), poly-L-lactic acid-co-glycolic acid (PLGA), PDLLA containing bioactive glass, PLGA containing bioactive glass, poly-L-lactic acid containing β-tricalcium phosphate (PLLA-TCP), poly-L-lactic acid containing hydroxyapatite (PLLAHA), polydioxanone (PDS), polyethylene glycol (PEG), poly(8-caprolactone) (PCL), polycaprolactone containing alginic acid (PCL), polyhydroxybutyrate (PHB), polycarbonate (PC), N-vinylpyrrolidone copolymer, polyorthoester, chitosan, poly(2-hydroxyethyl-methacrylate) (PHEMA), hyaluronic acid, and hydrogels.

[0306] These methods described herein are not by any means exhaustive, and additional methods suitable for a particular application will be apparent to those of skill in the art. Further, the effective amount of the composition can be further approximated by analogy to compounds known to exert the desired effect.

Examples

[0307] The present invention will be described in more detail by reference to the following examples. These examples are provided for illustrative purposes only and are not intended to be limiting, unless otherwise specified. Accordingly, the present invention should in no way be construed as being limited to the following examples, but rather should be construed to encompass any and all variations that become apparent as a result of the teachings provided herein.

[0308] Without further elaboration, it is believed that one of ordinary skill in the art can, using the preceding description and the following illustrative examples, make and utilize the present invention and practice the claimed methods. Accordingly, the following examples are specifically pointed out as preferred embodiments of the present invention and should in no way be construed as limiting the remainder of the disclosure.

[0309] Example 1: 4-Aminopyridine for the Treatment of Tissue Dysfunction Caused by Toxic Injury This example shows an in vivo study of properties associated with multiple toxic injuries. Specifically, an initial study was conducted on a specific case of CIPN induced by exposure to PTX. Figure 1 (including Figures 1A to 1H) shows representative results indicating that 4-aminopyridine (4-AP) exerted a protective effect against the development of chemotherapy-induced peripheral neuropathy (CIPN) and mitochondrial damage caused by clinically relevant doses of PTX. Sixteen-week-old C7BL / 6 female mice were administered a single dose of 35 mg / kg PTX (intraperitoneally). The animals were divided into a control (saline, intraperitoneally, n = 4) or treatment group (2 mg / kg 4-AP daily, intraperitoneally, n = 4). The Catwalk score before treatment was compared 21 days after PTX injection.

[0310] Figure 1A shows a representative nerve conduction test that demonstrated a significant increase in latency in the control group over the course of the test. In the 4-AP treated mice, the increase in latency was significantly smaller. Figure 1B shows a representative nerve conduction test that demonstrated a significant decrease in velocity in the control group over the course of the test. In the 4-AP treated mice, the decrease in velocity was significantly smaller. Figure 1C shows a representative Catwalk analysis that showed a significant increase during the swing phase in control mice, but no significant change in the 4-AP treated group (p<0.05, p<0.01, p<0.001). Figure 1D shows a representative Catwalk analysis that showed a significant increase during the stance phase in control mice (an indicator of gait impairment), but no significant change in the 4-AP treated group (*p<0.05, **p<0.01, ***p<0.001).

[0311] For clinical treatment, mice were treated with a PTX dose of 35 mg / kg (corresponding to approximately 110 mg / m 2 and within the clinical treatment range) over 4 cycles at 3-week intervals. From the results of this preliminary study (n = 4), it was shown that the animals tolerated the dose and that the concentration was sufficient to induce clinically significant signs of CIPN (Figure 1). Next, different cohorts of mice were treated with 35 mg / kg of PTX for 1 cycle (3 weeks), and this was also found to cause CIPN.

[0312] This study also included a 4-AP treatment group (n = 4), which received 4-AP intraperitoneally at a daily dose of 2 mg / kg (as in most studies by MN and co-workers). In the 4-AP group, it was found that the signs of CIPN were significantly reduced compared to control mice that received an intraperitoneal injection of an equal volume of saline. In the study, a low dose of 4-AP corresponding to approximately 40% of the mouse body surface area was used, which is equivalent to a dose of 20 mg / day used in the treatment of multiple sclerosis and is less than the equivalent dose tested in patients with chronic spinal cord injury.

[0313] In these tests, it was found that 4-AP treatment improved the CIPN-related changes in motor function (by Catwalk analysis) and electrophysiological parameters caused by PTX treatment, and reduced the PTX-induced changes in mitochondrial size and health. To evaluate the detailed functional changes in walking, the CatWalk (trademark) (Noldus Information Technology, Wageningen, The Netherlands) method was implemented. Electrophysiological data related to nerve conduction tests were also recorded, and in animals treated with 4-AP, both latency and velocity were improved.

[0314] It is also shown herein that co-treatment with 4-AP prevents PTX-induced changes in peripheral nerve function associated with CIPN. Samples of the sciatic nerve were taken, embedded in resin, sectioned, and imaged using a transmission electron microscope. The images were analyzed to calculate markers of peripheral neuropathy, namely, axon number, fiber size, myelination (g-ratio), and circumferential irregularity (Figure 2). Mitochondria were evaluated to assess chemotherapy-induced damage, and the results showed that 4-AP improved myelination. Figure 2 (including Figures 2A to 2E) shows representative results indicating that 4-AP prevents chemotherapy-induced axonal damage. Figure 2A shows representative G-ratio (axon:myelin area) data indicating that axons within the 4-AP treatment are better myelinated than those in the control group. Figure 2B shows representative circularity data indicating that axons within the 4-AP treatment are also more regularly structured than those in the control group. Figure 2C shows a representative baseline appearance of the sciatic nerve. Figure 2D shows a representative appearance of control axons 3 weeks after a single treatment with 35 mg / kg of PTX. Figure 2E shows a representative appearance of axons treated with 4-AP (2 mg / kg daily), showing thicker myelin and a more regular structure.

[0315] Example 2 In a second test using a longer period of PTX administration, it further shows the ability to prevent the development of CIPN when 4-AP is co-administered.

[0316] All animals were intraperitoneally injected with 35 mg / kg of PTX four times at 3-week intervals (equivalent to approximately 110 mg / m 2 corresponding to, within the clinical treatment range, delivered in the form of a diluted solution of taxol used clinically). A subset of animals was simultaneously given 4-AP (1 mg / kg, daily), which was started at the same time as the first PTX treatment and continued overnight. The responses of functional outcomes were recorded at 3-week intervals throughout the study period and included von Frey monofilament testing, cold plate thermal sensitivity, CatWalk™ gait analysis, and nerve conduction testing. All animals had functionally demonstrable peripheral neuropathy at 6 weeks of age.

[0317] In these experiments, it was observed that co-treatment with 4-AP prevented the development of PTX-induced CIPN. 4-AP treatment prevented PTX-induced changes in multiple motor functions and electrophysiological parameters. Analysis of the response to thermal changes using cold plate analysis showed that 4-AP prevented the development of CIPN-like changes regardless of whether the changes indicated hyperalgesia or hypoalgesia. Analysis of mechanical allodynia using the von Frey filament test revealed an increase in sensitivity as indicated by a response to a lesser force. This change became apparent by 3 weeks, worsened at 6 weeks, and plateaued over the next 6 weeks. In contrast, mice treated with PTX+4-AP showed no change from the baseline of their response, and the differences from the PTX-alone group were statistically significant at all time points. A significant increase in sensitivity to cold, determined by the number of times the mice tried to remove their paws from the cold plate stimulus, was apparent at 6 weeks after the start of treatment and was maintained throughout the remaining 6-week analysis. In contrast, mice treated with PTX+4-AP showed no change from the baseline of this parameter. The differences between the two treatment groups were highly statistically significant at the 6, 9, and 12-week time points. Other responses to cold, determined by the number of times the mice lifted their paws from the cold plate or licked their hind paws, indicated a loss of sensitivity in PTX-treated mice. For these outcomes as well, co-treatment with 4-AP prevented the manifestation of these changes, and the differences between the PTX group and the PTX+4-AP group were statistically significant.

[0318] In these experiments, treatment with CIPN also decreased the amplitude and velocity of nerve impulse conduction, and large changes were seen 6 weeks after the start of PTX treatment and continued for the following 6 weeks. For both of these parameters, co-treatment with 4-AP prevented the manifestation of these changes, and the differences between the PTX group and the PTX+4-AP group were statistically significant.

[0319] Example 3: Treatment with 4-AP prevents PTX-induced CIPN caused by repeated PTX exposure, even when applied at very low levels of 4-AP. When repeated treatment with PTX is performed, it is important to know whether 4AP treatment can also prevent PTX-induced peripheral neuropathy. This is a situation that will occur clinically where cancer patients are not treated with a single dose of paclitaxel but instead receive repeated exposure. Whether 4AP is useful in preventing the effects of this abnormal type of injury, which has elements of repetitive damage due to repeated exposure to paclitaxel and elements of chronic damage due to the accumulation of the resulting damage, has not been shown heretofore. Thus, in this situation, 4-AP needs to prevent further damage in order to overcome existing damage and improve chemotherapy-induced changes.

[0320] To examine whether 4-AP also confers a protective effect against the development of CIPN when animals are repeatedly exposed to PTX, 16-week-old C7BL / 6 female mice were given 4 cycles of PTX treatment at a dose of 35 mg / kg PTX (intraperitoneally). The animals were divided into a control (saline, intraperitoneally) group used in Examples 1 and 2 or a treatment group treated with a 25% dose of 4AP (i.e., 0.5 mg / kg of 4-AP daily, intraperitoneally). The pre-treatment Catwalk score was compared 21 days after PTX injection.

[0321] As a result of the analysis, application of the von Frey filament test revealed that 4-AP treatment reduced PTX-induced mechanical allodynia, in which case the sensitivity to painful stimuli increased as the sensitivity to smaller filaments was shown. The outcomes were very significant at 3, 6, 9, and 12 weeks. 4-AP treatment also prevented the development of symptoms opposite to hypoalgesia using analysis of foot lifting and jumping behavior in response to cold plate stimulation. The outcome of foot lifting showed a significant trend at 6 and 9 weeks and was very significant at 12 weeks. The jumping behavior was very significant at 6, 9, and 12 weeks (Figures 3A - 3C).

[0322] 4-AP treatment also improved PTX-induced gait impairment detected by Catwalk analysis, resulting in a significant effect at multiple time points (Figures 3D - 3F).

[0323] The 4-AP treatment also improved PTX-induced nerve conduction abnormalities in multiple outcomes of latency (Figure 3G), amplitude (Figure 3H), and velocity (Figure 3I).

[0324] Example 4: The 4-AP treatment prevents changes in the structure of PTX-induced peripheral nerves caused by repeated PTX exposure, even when applied at very low levels of 4-AP. It is important to know whether 4AP treatment can also prevent PTX-induced peripheral neuropathy when repeated treatment with PTX is performed. This is a situation that will occur clinically where cancer patients are not treated with a single dose of paclitaxel but instead receive repeated exposure. Nothing has been shown so far regarding whether 4AP is useful in preventing the effects of this abnormal type of injury, which has elements of repetitive injury due to repeated exposure to paclitaxel and elements of chronic injury due to the accumulation of the resulting damage.

[0325] It is also particularly important to determine whether such treatment prevents the structural damage associated with the development of CIPN. The ability of 4-AP treatment to prevent damage caused by anticancer agents that affect cells in many different ways is not yet known. It is known that significant structural damage occurs with chemotherapy treatment, but from current information, it is impossible to predict whether 4AP has any effect in preventing this type of damage.

[0326] To determine whether 4-AP prevents chemotherapy-induced structural damage even when chemotherapy is applied multiple times, ultrastructural analysis of tissues from mice treated with 4 cycles of 35 mg / kg of PTX every 3 weeks was performed. A low dose of 4-AP, only 25% of the dose used in Examples 1 and 2, was used, and this was applied at 0.5 mg / kg per day instead of 2 mg / kg per day.

[0327] Superstructural analysis of the myelination performed revealed multiple advantages of 4-AP treatment (Figure 4). Analysis of the G ratio (axon:myelin area) data showed that axons in 4-AP treated mice were better myelinated than those in the control (PTX + saline) group (Figure 4C). Analysis of circularity also revealed that axons in 4-AP treatment were also more regularly structured than those in the control group (Figure 4D). In the 4-AP treatment group, the frequency of myelin contour degeneration also decreased.

[0328] These experiments also revealed that treatment with 4AP improves mitochondrial health as determined by mitochondrial structure even under conditions of repeated exposure to chemotherapy.

[0329] Example 5: 4-AP treatment is effective in recovering from PTX-induced CIPN even after peripheral neuropathy has been established, at both the functional and histological analysis levels Another important issue to consider is whether treatment with 4AP can be delayed after CIPN has been established. This is a situation relevant to ongoing chemotherapy treatment and is clinically very important because peripheral neuropathy can be a dose-limiting factor in cancer treatment. If CIPN develops, patients may defer necessary treatment due to the impact of CIPN on quality of life.

[0330] Treatment with chemotherapy constitutes repeated injury, which starts with the first treatment and continues to increase with subsequent treatments, and nothing has been shown beforehand about the effect of 4AP in this situation. Prior studies on the use of 4AP in the treatment of established syndromes such as multiple sclerosis and spinal cord injury have shown limited results in this regard. As discussed elsewhere, the effect of 4AP, especially on the relief of pain-related symptoms, is likely to be at best limited and more likely to be ineffective.

[0331] To investigate this problem, 16-week-old C7BL / 6 female mice were given 4 cycles of PTX treatment at a dose of 35 mg / kg PTX (intraperitoneally). The animals were divided into treatment groups treated with control (saline, ip), or a 25% dose of 4-AP used in Figures 1 and 2 (i.e., 0.5 mg / kg of 4-AP per day, intraperitoneally, starting 6 weeks after the first PTX treatment and at the time when CIPN-related changes were already apparent).

[0332] These experiments revealed that 4-AP treatment, which was initiated 6 weeks after the mice received the third exposure to PTX, also restored the mice from the effects of repeated PTX treatment on the peripheral nerves. For example, by applying the von Frey filament test (sensitivity to smaller filaments indicates an increase in sensitivity to painful stimuli), testing the 4-AP treatment for PTX-induced mechanical hypersensitivity revealed that the benefits of 4-AP treatment began to be observed at week 12 (Figure 5A). 4-AP treatment also restored the mice from hypersensitivity symptoms to heat stimuli using the analysis of jump behavior in response to cold plate stimuli (Figure 5B). The changes in jump behavior were significant at weeks 9 and 12.

[0333] 4-AP treatment also restored the mice from PTX-induced gait impairment detected by Catwalk analysis, and significant effects on swing time and regularity index were observed at week 9 (the first analysis after the start of 4-AP treatment) and week 12, and the outcome of stance time also showed a significant trend at these same time points (Figures 5C - 5E).

[0334] 4-AP treatment also restored 4-AP from PTX-induced nerve conduction abnormalities in multiple outcomes of latency (Figure 5F), amplitude (Figure 5G), and velocity (Figure 5H) after PTX-induced nerve conduction abnormalities were established by multiple PTX exposures.

[0335] 4-AP treatment also restored the mice from PTX-induced structural myelin abnormalities showing myelin degeneration (Figure 5I). This result reveals an unexpected ability to promote tissue repair after CIPN has already been established.

[0336] Example 6: 4-AP treatment continuously restores PTX-induced CIPN even after peripheral neuropathy has been established, and the effect persists after treatment is discontinued. One of the important problems in the treatment of injuries induced by exposure to toxic insults is the need for a treatment that can reverse already incurred damage. Many people are not diagnosed with an injury until after it has occurred. Further, in certain cases of peripheral neuropathy caused by exposure to chemotherapeutic agents, neuropathy can manifest after treatment has been discontinued. Additionally, it may be desirable to delay the initiation of neuropathy treatment until after cancer treatment is complete.

[0337] Regarding treatment of injury with 4-AP, there is no prior information that would enable prediction of the outcome of established injury. In fact, predictions based on years of research on chronic injury do not predict such an effect. This is because in syndromes such as multiple sclerosis, chronic spinal cord injury, spinocerebellar ataxia, and other established pathologies where 4AP has been reported to have symptomatic relief, the effects of 4AP treatment disappear when treatment is discontinued.

[0338] Therefore, the question of whether treatment of CIPN with 4-AP can promote recovery that is maintained after treatment completion is unpredictable. This unpredictability is further exacerbated by the fact that chemotherapy treatment itself is a repetitive insult with established chronic elements that worsen with each additional treatment. This is a situation that is very different from previous analyses of the effects of 4-AP treatment. Nevertheless, prior studies on established injury predict that no permanent recovery will occur if treatment with 4-AP is initiated more than a few weeks after the initial injury has occurred. Thus, not only is there insufficient data indicating whether 4-AP has any effect on the treatment of CIPN, but it is not possible to predict whether the effects that actually occur will persist after treatment is discontinued, i.e., in situations where tissue repair will be required for the treatment of established injury.

[0339] To investigate this problem, 16-week-old C7BL / 6 female mice were given 4 cycles of PTX treatment at a dose of 35 mg / kg PTX (intraperitoneally). All animals exhibited functionally demonstrable peripheral neuropathy at 6 weeks. The animals were then treated with the same dose and treatment regimen as used in Figure 5. At 12 weeks after the first exposure to PTX and 6 weeks after treatment with 4-AP, the 4-AP treatment was terminated, and the animals were observed for an additional 6 weeks (i.e., 6 weeks without 4-AP treatment) after this time point.

[0340] These experiments revealed the unexpected result that 4-AP treatment can restore and maintain the effects on peripheral nerve function caused by repeated exposure to PTX, even after treatment has ended. In contrast to the predicted 99.9% excretion of 4-AP in 24 hours (by renal clearance), the effect was still present 6 weeks after the end of treatment. Such persistent changes not only result in symptomatic relief that only exists during the treatment period but also indicate a restorative promoting effect of 4-AP treatment.

[0341] The results of these experiments show that 4-AP treatment can restore normal function several weeks after PTX treatment, regardless of whether the symptoms are hyperalgesia or hypoalgesia, which is the most promising among all outcomes in addressing clinical needs. Figure 6A shows the effect of 4-AP treatment on PTX-induced mechanical hypersensitivity tested by the application of von Frey filament tests, indicating that the sensitivity to painful stimuli increases as the sensitivity to smaller filaments is shown. The effect began to be observed at 12 weeks and was maintained at 15 weeks and 18 weeks. Figure 6B shows that 4-AP treatment can also cause a persistent recovery from the opposite symptoms of decreased sensitivity to stimuli using the analysis of foot withdrawal (Figure 6B) and jump behavior (Figure 6C) in response to cold plate stimuli.

[0342] Figure 6D shows the ability of 4-AP to continuously recover from PTX-induced gait impairment as determined by the Catwalk Regularity Index. The effect was observed at week 9 (i.e., 3 weeks after 4-AP treatment) and was maintained for at least 6 weeks after the end of 4-AP treatment. Figure 6E shows the ability of 4-AP to recover their latencies after PTX-induced nerve conduction abnormalities were established by multiple PTX exposures.

[0343] Figures 6F-6G show the ability of 4-AP to bring about persistent reparative changes when used to treat established PTX-induced CIPN and tissue damage. Changes in the G ratio (Figure 6F) and circularity (Figure 6G) caused by repeated PTX exposures recovered to normal at the 12-week time point, and these effects were maintained at the 18-week time point (i.e., 6 weeks after the end of treatment).

[0344] Example 7: 4-AP treatment restores cisplatin-induced CIPN Determine whether the effects provided by the 4-AP treatment experiments were demonstrated with another toxic substance having a different mechanism of action from paclitaxel. In these experiments, the toxic agent tested was cisplatin. In contrast to the microtubule-stabilizing activity of taxanes (including paclitaxel), the platinum-containing compound cisplatin is thought to kill rapidly dividing cells by causing DNA cross-links. Paclitaxel and cisplatin also differ greatly in their structures. The difference in the mechanisms of action of cisplatin and paclitaxel is clarified by the fact that paclitaxel is often used in the treatment of cancers that are resistant to cisplatin. Thus, this example describes in vivo tests of properties related to multiple toxic injuries.

[0345] In these experiments, 16-week-old C7BL / 6 female mice were treated with CIS (2.5 mg / kg, once a week, intraperitoneally) for 8 weeks. The animals were then treated with 4-AP (1 mg / kg per day) for the next 6 weeks (n = 4).

[0346] These experiments revealed that the 4-AP-treated mice that started the treatment 9 weeks after the start of CIS treatment recovered normal body weight. The CIS-treated mice developed CIPN, and 4-AP was effective in restoring PTX-induced CIPN, similar to restoring CIS-induced CIPN. Analysis of hyperalgesia by von Frey filament analysis showed that sensitivity increased dramatically in CIS-treated mice and recovered to normal levels with 4-AP treatment. The recovery of normal sensitivity was persistent and maintained for at least 2 weeks after the end of treatment, far exceeding the 12 - 16 hours when substantially all of the 4-AP was expected to be removed from the body, thus showing a sustained recovery of function and a repair-promoting effect of the treatment. Figure 7C shows representative results demonstrating the effect of 4-AP on nerve impulse amplitude, which also recovered with 4-AP treatment and the improvement was maintained for at least 2 weeks after the end of treatment with 4-AP. Figure 7D shows representative results demonstrating the effect of 4-AP on nerve impulse velocity, which also recovered with 4-AP treatment and the improvement was maintained for at least 2 weeks after the end of treatment with 4-AP (in contrast to the prediction that more than 99.9% of 4-AP would be excreted (by renal clearance) in 24 hours), thus showing a sustained recovery of function and a repair-promoting effect of the treatment. This outcome could only be explained by 4-AP-induced reparative changes in the damaged tissue.

[0347] Example 8: 4-AP treatment prevents chemotherapy-induced damage to the kidney Another tissue that is often damaged by treatment with chemotherapeutic agents is the kidney, and it is very important to provide protection from such nephrotoxicity. Combining focusing on the kidney with the fact that exposure to toxic injury such as chemotherapeutic agents is a very different type of injury from traumatic injury and the injuries that occur with other uses of 4-AP that have been tested so far, it cannot be predicted whether the kidney is also included in the effects of 4-AP with respect to the present invention.

[0348] To examine the effects on the kidney, histological tests were performed on this tissue. Mice were given PTX on day 0 of the experiment, and half of those mice were also injected with 4-AP at a dose of 2 mg / kg daily at this time point and thereafter. Mice not given 4-AP were injected with saline to control for any stress induced by daily injections. The experimental groups also included mice not receiving PTX treatment and mice treated only with daily injections of saline. After one week, the mice were sacrificed. The mice were perfused with 4% paraformaldehyde, the kidneys were sectioned, and stained with hematoxylin and eosin (H&E).

[0349] H&E staining of the kidneys treated with saline, PTX, and PTX + 4-AP revealed normal proximal and distal tubules in both the saline- and PTX + 4-AP-treated kidneys (Figure 8). In the PTX-treated kidneys, loss of the brush border (dark gray arrow), nuclear dropout (gray arrow), and vacuolization of the tubular epithelial cells (black arrow) were observed, but not in the kidneys isolated from animals treated with PTX + 4-AP.

[0350] The results of this experiment showed that co-treatment with 4-AP also prevented multiple aspects of histological kidney damage caused by PTX exposure. This surprising result indicates that protection against chemotherapy via 4-AP is also seen in tissues that are not part of the nervous system. These results further support the data shown in Table 1.

[0351] Example 9: 4-AP exposure prevents PTX-induced changes in the central nervous system Another tissue that is important in damage due to anticancer agents, radiation, and other forms of toxic injury is the central nervous system (CNS). The CNS is affected by many anticancer treatments, radiation, and many toxic injuries. Such damage can lead to changes in multiple neurological and / or cognitive functions, and preventing such damage is of great medical importance. Interventions to prevent such damage are far less available than for other parts of the body.

[0352] One of the first signs of damage to the CNS is an increase in the expression of glial fibrillary acidic protein (GFAP). The increase in GFAP indicates astrocyte activation and increases in response to chemical damage such as inflammation, various physical injuries, and exposure to chemotherapeutic agents (e.g., Liu, et al., Paclitaxel-activated astrocytes produce mechanical allodynia in mice by releasing tumor necrosis factor-α and stromal-derived cell factor 1. J.Neuroinflammation 2019;16:209; Masocha, Astrocyte activation in the anterior cingulate cortex and altered glutamatergic gene expression during paclitaxel-induced neuropathic pain in mice. PeerJ 2015 3:e1350).

[0353] Sixteen-week-old female C57BL / 6 mice were inoculated with the triple-negative mouse breast cancer cell line E0771 to determine whether 4-AP treatment could prevent the PTX-induced increase in the reactive inflammatory response in the brain detected by an increase in the expression of glial fibrillary acidic protein (GFAP). When the tumors were palpable, mice were injected with paclitaxel and water, 4-AP, both, or saline as a control. Paclitaxel (35 mg / kg) was injected on day 1, and 4-AP and water were injected daily. On day 9, the mice were sacrificed, and their brains were perfused and harvested. The brains were cryosectioned coronally.

[0354] Mouse brain sections were stained with anti-GFAP antibody, followed by fluorescence secondary staining and analyzed by immunofluorescence. Images of the corpus callosum, a major myelinated tract in the CNS, were obtained via a confocal microscope and analyzed with Image-J. In these experiments, it was revealed that PTX treatment was associated with an increase in GFAP expression (Figure 9). In mice co-treated with 4-AP, the increase in GFAP expression induced by PTX exposure was significantly reduced.

[0355] Example 10: The protection by 4-AP against the toxicity of chemotherapeutic agents is selective for normal cells One further unexpected property of 4-AP is that its ability to provide protection from chemotherapeutic agents with different mechanisms of action is selective for normal tissues (i.e., non-transformed cells), and 4-AP does not provide protection against cancer cells. Since many survival mechanisms are the same in normal and cancer cells, this outcome was also unpredictable. Protecting cancer cells would significantly reduce the value of 4-AP as a therapeutic agent in cancer treatment, so this surprising result is very important. Furthermore, many of the toxic insults relevant to this patent (including chemotherapeutic agents, radiation, and pesticides) increase the risk of cancer, so protection of cancer cells would also limit the use of 4-AP.

[0356] To examine the effect of 4-AP treatment on cancer cells, 4-AP was combined with paclitaxel or cisplatin, and dose-response curves spanning more than 20-fold ranges were drawn for the widely studied E0771 mouse breast cancer cell line and A549 human lung cancer cell line. Cells were exposed to paclitaxel or cisplatin for 5 days in the presence or absence of 1 mM 4-AP. This concentration of 4-AP is typically used in in vitro tests and far exceeds concentrations achievable in vivo. This higher dose would increase the likelihood of detecting protective activity.

[0357] Figure 10 shows that the presence of 4-AP does not protect cancer cells from cisplatin, but rather may enhance sensitivity to low doses of paclitaxel. Thus, this example reveals the surprising result that the ability of 4-AP to protect multiple types of normal tissues from the toxic effects of cisplatin and paclitaxel is abnormally selective and not applicable to cancer cells.

[0358] Example 11. 4-AP treatment prevents changes in renal function as revealed by analysis of serum 4-AP levels. Furthermore, Table 1 shows data indicating that circulating 4-AP levels analyzed 1 hour after intraperitoneal injection were not different between mice administered 4-AP alone and mice administered 4-AP + PTX, indicating that 4-AP prevented PTX-induced changes in renal function. After the start of treatment, 4-AP was intraperitoneally injected at the indicated time points. Mice were given PTX on day 0 of the experiment, and half of those mice were also injected with 4-AP at a dose of 1 g / kg daily at this time point and thereafter. Blood samples were taken 3 and 7 days after the start of the experiment and also 1 hour after injection of 4-AP. Circulating levels of 4-AP were measured by HPLC analysis. As shown, the amount of 4-AP detected in circulation was not different between mice treated with 4-AP alone or PTX + 4-AP, whether on day 3 or day 7. Since 4-AP is excreted by renal clearance, this further indicated that 4-AP prevented PTX-induced changes in renal function. [Table 1]

[0359] The disclosures of all patents, patent applications, and publications cited herein are hereby incorporated by reference in their entirety. Although the invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and variations of the invention may be devised by those skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

Claims

Claim 1 A method for preventing, alleviating, or treating tissue damage or tissue dysfunction caused by toxic injury in a subject in need thereof, wherein the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker or an analog thereof, a racemic compound, a tautomer, an isomer, an enantiomer, a diastereomer, a prodrug, a pharmaceutically acceptable salt, or a derivative thereof, said method for prevention, alleviation, or treatment. Claim 2 A method for reducing or recovering tissue damage, oxidative damage, scarring, or any combination thereof in a subject in need thereof, wherein the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker or an analog thereof, a racemic compound, a tautomer, an isomer, an enantiomer, a diastereomer, a prodrug, a pharmaceutically acceptable salt, or a derivative thereof, wherein the tissue damage, oxidative damage, scarring, or any combination thereof is caused by toxic injury, said method for reduction or recovery. Claim 3 A method for recovering, improving, or enhancing at least a part of tissue function in a subject in need thereof, wherein the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker or an analog thereof, a racemic compound, a tautomer, an isomer, an enantiomer, a diastereomer, a prodrug, a pharmaceutically acceptable salt, or a derivative thereof, wherein the tissue function is reduced by toxic injury, said method for recovery, improvement, or enhancement. Claim 4 The method according to claim 3, wherein the tissue function is motor function, sensory function, cognitive function, visual function, auditory function, renal function, hematopoietic function, normal skin function, salivary gland function, liver function, gallbladder function, gastrointestinal (GI) function, sexual function, or any combination thereof. Claim 5 A method for recovering, improving, or enhancing myelination in a subject in need thereof, wherein the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker or an analog thereof, a racemic compound, a tautomer, an isomer, an enantiomer, a diastereomer, a prodrug, a pharmaceutically acceptable salt, or a derivative thereof, wherein myelination is reduced or regulated by toxic injury, said method for recovery, improvement, or enhancement. Claim 6 A method for preventing, alleviating, or treating mitochondrial damage or mitochondrial dysfunction caused by toxic injury in a subject in need thereof, comprising: The method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker or an analog thereof, a racemic compound, a tautomer, an isomer, an enantiomer, a diastereomer, a prodrug, a pharmaceutically acceptable salt, or a derivative thereof, said method of prevention, alleviation, or treatment. **Claim 7** A method for preventing, alleviating, or treating axonal damage or axonal dysfunction caused by toxic injury in a subject in need thereof, comprising: The method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker or an analog thereof, a racemic compound, a tautomer, an isomer, an enantiomer, a diastereomer, a prodrug, a pharmaceutically acceptable salt, or a derivative thereof, said method of prevention, alleviation, or treatment. **Claim 8** A method for preventing, alleviating, or treating at least one gait disorder caused by toxic injury in a subject in need thereof, comprising: The method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker or an analog thereof, a racemic compound, a tautomer, an isomer, an enantiomer, a diastereomer, a prodrug, a pharmaceutically acceptable salt, or a derivative thereof, said method of prevention, alleviation, or treatment. **Claim 9** A method for enhancing tissue regeneration, cell survival, or a combination thereof in a subject in need thereof, comprising: The method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker or an analog thereof, a racemic compound, a tautomer, an isomer, an enantiomer, a diastereomer, a prodrug, a pharmaceutically acceptable salt, or a derivative thereof, wherein the subject has been exposed to at least one toxic injury, said method of enhancement. **Claim 10** The method according to claim 9, wherein the tissue is kidney tissue, liver tissue, heart tissue, lung tissue, brain tissue, central nervous system tissue, peripheral nerve tissue, gastrointestinal tract tissue, intestinal tissue, visual system tissue, auditory system tissue, skin tissue, bladder tissue, reproductive system tissue, hematopoietic system tissue, musculoskeletal tissue, or any combination thereof. **Claim 11** A method for preventing, alleviating, or treating chemotherapy-induced peripheral neuropathy (CIPN) in a subject in need thereof, wherein the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one potassium channel blocker or an analog thereof, a racemic compound, a tautomer, an isomer, an enantiomer, a diastereomer, a prodrug, a pharmaceutically acceptable salt, or a derivative thereof. **Claim 12** The method according to claim 11, wherein the chemotherapy-induced peripheral neuropathy (CIPN) is CIPN caused by a taxane agent (P-CIPN), CIPN caused by cisplatin treatment (CisIPN), CIPN caused by an anticancer agent, CIPN caused by a platinum-based antitumor agent, CIPN caused by a vinca alkaloid agent, CIPN caused by an epothilone agent, CIPN caused by a proteasome inhibitor, CIPN caused by an immunomodulator, or any combination thereof. **Claim 13** The method according to any one of claims 1 to 12, wherein the at least one potassium channel blocker comprises 4-aminopyridine, a derivative of 4-aminopyridine, or a combination thereof. **Claim 14** The derivative of 4-aminopyridine is of formula (I) 【Chemical 1】 or a compound having a structure of an analog thereof, a racemic compound, a tautomer, an isomer, an enantiomer, a diastereomer, a prodrug, a pharmaceutically acceptable salt, or a derivative. In the formula, R １ , R ２ , R ３ , R ４ , and R ５ are each independently selected from hydrogen, halogen, C １ -C ６ alkyl, amine, hydroxyl, alkoxy, carboxyl, or any combination thereof, wherein R １ , R ２ , R ３ , R ４ , and R ５ are optionally substituted, the method according to claim 13. **Claim 15** The tissue damage is kidney tissue damage, liver tissue damage, heart tissue damage, lung tissue damage, brain tissue damage, central nervous system damage, peripheral nerve tissue damage, peripheral neuropathy, nephrosis, chemotherapy-induced peripheral neuropathy (CIPN), radiation-induced peripheral neuropathy (RIPN), chemotherapy-induced nephrotoxicity (CINT), chemotherapy-induced neutropenia, radiation-induced neutropenia, gastrointestinal tissue damage, intestinal tissue damage, visual system tissue damage, auditory system tissue damage, skin tissue damage, bladder tissue damage, reproductive system tissue damage, hematopoietic system tissue damage, or any combination thereof. The method according to any one of claims 1, 2, or 14. **Claim 16** The method according to claim 15, wherein the chemotherapy-induced peripheral neuropathy (CIPN) is CIPN caused by a taxane agent (P-CIPN), CIPN caused by cisplatin treatment (CisIPN), CIPN caused by an anticancer agent, CIPN caused by a platinum-based antitumor agent, CIPN caused by a vinca alkaloid agent, CIPN caused by an epothilone agent, CIPN caused by a proteasome inhibitor, CIPN caused by an immunomodulator, or any combination thereof.

17. The method according to any one of claims 1, 2, or 14, wherein the tissue damage is multi-tissue damage, multi-organ tissue damage, or any combination thereof.

18. The method according to any one of claims 1 to 14, wherein the toxic injury is acute toxic injury, chronic toxic injury, or any combination thereof.

19. The method according to any one of claims 1 to 14, wherein the toxic injury includes exposure to radiation at a damage level along the electromagnetic spectrum.

20. The method according to any one of claims 1 to 14, wherein the toxic injury is a non-biological substance, a non-natural compound, a toxin, a drug used in the treatment of cancer, a chemotherapeutic agent, a biological response modifier, radiation, or any combination thereof.

21. The method according to claim 20, wherein the chemotherapeutic agent includes a platinum-based antitumor agent, a vinca alkaloid agent, an epothilone agent, a taxane agent, a proteasome inhibitor, an immunomodulator, or any combination thereof.

22. The method according to claim 20, wherein the toxic injury includes at least one environmental toxicant.

23. The method according to claim 20, wherein the radiation includes radiation from radioactive cancer treatment, radiation from a nuclear energy accident, radiation from exposure to nuclear waste, radiation from the use of radioactive substances in military applications, or any combination thereof.

24. The toxic injury includes at least a first toxic injury and a second toxic injury, the first toxic disorder is a chemotherapeutic agent, radiation, or a combination thereof, and the second toxic disorder is a chemotherapeutic agent, radiation, or a combination thereof. The method according to any one of claims 1 to 14.

25. The method according to any one of claims 1 to 14, wherein the therapeutically effective amount of the pharmaceutical composition is administered to the subject before the toxic injury.

26. The method according to any one of claims 1 to 14, wherein the therapeutically effective amount of the pharmaceutical composition is administered to the subject at the time of the toxic injury.

27. The method according to any one of claims 1 to 14, wherein the therapeutically effective amount of the pharmaceutical composition is administered to the subject after the toxic injury.

28. The method according to any one of claims 1 to 14, wherein the therapeutically effective amount of the pharmaceutical composition is administered to the subject in multiple doses.

29. The method according to any one of claims 1 to 14, wherein the therapeutically effective amount of the pharmaceutical composition is repeatedly administered to the subject over a period of about 1 day to about 100 years.

30. The method according to any one of claims 1 to 14, wherein the therapeutically effective amount of the pharmaceutical composition is administered to the subject systemically, locally, or in a combination thereof.

31. The method according to any one of claims 1 to 14, wherein the therapeutically effective amount of the pharmaceutical composition is administered to the subject by intraperitoneal injection, intravenous injection, intramuscular injection, intrathecal injection, subcutaneous injection, sublingual administration, inhalation, oral administration, transdermal administration, administration to the exterior of the body in the form of a liquid, administration to the exterior of the body in the form of an ointment, administration to the exterior of the body in the form of a dressing, or any combination thereof.

32. The method according to any one of claims 1 to 14, wherein the therapeutically effective amount of the pharmaceutical composition is administered to the subject at a dose of a potassium channel blocker of about 1 mg / day to about 1,000 mg / day.

33. The method according to claim 32, wherein the therapeutically effective amount of the pharmaceutical composition is administered to the subject at a dose of a potassium channel blocker of about 2.5 mg / day to about 40 mg / day.

34. The method according to claim 32, wherein the therapeutically effective amount of the pharmaceutical composition is administered to the subject at a dose of a potassium channel blocker of about 40 mg / day to about 100 mg / day.

35. The method according to any one of claims 1 to 14, wherein the therapeutically effective amount of the pharmaceutical composition is co-administered with at least one anti-seizure agent or a composition thereof.

36. The method according to claim 35, wherein the at least one anticonvulsant is barbiturate, benzodiazepine, bromide, carbamate, carboxamide, fatty acid, fructose or its derivative, γ-aminobutyric acid (GABA) or its analog, hydantoin, oxazolidinedione, propionate, pyrimidinedione, pyrrolidine, succinimide, sulfonamide, triazine, urea, valproylamide, or any combination thereof.

37. The method according to any one of claims 1 to 36, wherein the method further results in enhancing the repair or regeneration of endogenous stem cells, enhancing the repair or regeneration of transplanted stem cells, enhancing the repair or regeneration of progenitor cells, promoting neurogenesis, enhancing cell survival, reducing scarring, reducing the size of lesions, reducing oxidative damage, or any combination thereof.

38. A method for identifying a subject responsive to administration of 4-aminopyridine for preventing, alleviating, or treating tissue damage or tissue dysfunction caused by toxic injury in a subject in need thereof, wherein the method comprises a) administering to the subject 1 to 5 times a therapeutically effective amount of a pharmaceutical composition comprising 4-aminopyridine, a derivative of 4-aminopyridine, or a combination thereof; b) evaluating in the subject the symptoms of tissue damage or tissue dysfunction caused by the toxic injury; c) identifying the subject as responsive to administration of 4-aminopyridine for preventing, alleviating, or treating the tissue damage or the tissue dysfunction caused by the toxic injury when the symptoms of the tissue damage or the tissue dysfunction caused by the toxic injury in the subject are improved.

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