Composition for Intermittent Administration of Calcineurin Inhibitor

Abstract

This specification provides compositions and dosing regimens of calcineurin inhibitors and cytochrome p450 inhibitors. The present invention provides, for example, a method of treating an allogeneic immune, autoimmune, inflammatory, or mitochondrial condition in a subject, the method comprising administering a calcineurin inhibitor and a cytochrome p450 inhibitor to the subject according to an intermittent dosing schedule. In one embodiment, the calcineurin inhibitor and the cytochrome p450 inhibitor are administered once a week.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims the priority of U.S. Provisional Application No. 63 / 359,350, filed on July 8, 2022, and U.S. Provisional Application No. 63 / 456,017, filed on March 31, 2023, the entire contents of each of which are incorporated herein by reference.

[0002] The present disclosure further provides a method of intermittently administering (e.g., once a week) a calcineurin inhibitor (e.g., cyclosporine) in combination with a cytochrome p450 enzyme inhibitor (e.g., ritonavir) to alleviate, prevent the onset of, or delay the onset of autoimmune, alloimmune, inflammatory, and / or mitochondrial conditions. In some embodiments, the methods and compositions described herein are useful for alleviating, delaying the onset of, or preventing the onset of autoimmune, alloimmune, and inflammatory conditions associated with atopy, perianal fistulas, canine exfoliative pemphigus, sebaceous adenitis, granulomatous meningoencephalitis, inflammatory bowel disease, or stomatitis.

Background Art

[0003] Calcineurin inhibitors are used as treatments for various alloimmune, autoimmune, and inflammatory conditions in both humans and non - human animals. Such agents have also been shown to be promising as treatments for mitochondrial dysfunction - related conditions through their action on mitochondrial mass flux [Fournier, et al.]. However, the use of such agents is limited by the frequency of their associated adverse effects and the frequency of administration of the dose required in patients [Azzi, et al.].

Summary of the Invention

Means for Solving the Problems

[0004] Calcineurin inhibitors are often used as treatments for conditions related to the immune system, such as, for example, cyclosporine for psoriasis [Ellis et al.] or tacrolimus for preventing rejection in liver transplantation [Haddad, et al.]. However, such treatments are limited by the adverse effects associated with calcineurin inhibitors and the frequency of dosing required for the patient. In cats, cyclosporine, for example, is associated with nausea and must be administered at least once a day [Roberts, et al.], while tacrolimus is associated with anemia and also must be administered twice a day [Kyles, et al.]. Thus, there is a need for new calcineurin inhibitor formulations that can alleviate such undesirable conditions.

[0005] Calcineurin inhibitors are metabolized via cytochrome p450 enzymes. Without being bound by theory, the present disclosure includes the insight that an inhibitor of cytochrome p450 can, when administered concurrently with a calcineurin inhibitor, increase the half-life of the calcineurin inhibitor in the body and decrease the rate of decline of the level of the calcineurin inhibitor in the bloodstream [Dresser et al.].

[0006] In one aspect, the present disclosure provides a method for alleviating autoimmune, alloimmune, inflammatory, and mitochondrial conditions, the method comprising administering to a subject or biological sample a calcineurin inhibitor or a pharmaceutically acceptable salt thereof and a cytochrome p450 inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the autoimmune, alloimmune, or inflammatory condition is associated with an increase in the level of a lymphokine or interleukin. In some embodiments, the mitochondrial condition is associated with the mitochondrial permeability transition pore (MPTP).

[0007] In another aspect, the present disclosure provides a method for preventing adverse effects associated with the metabolism of calcineurin inhibitors, the method comprising contacting calcineurin with a calcineurin inhibitor or a pharmaceutically acceptable salt thereof, and contacting cytochrome p450 with a cytochrome p450 inhibitor or a pharmaceutically acceptable salt thereof.

[0008] In some embodiments, the calcineurin inhibitor is selected from the group consisting of cyclosporine, tacrolimus, and pimecrolimus, and analogs or derivatives thereof. In some embodiments, the calcineurin inhibitor is cyclosporine.

[0009] In some embodiments, the calcineurin inhibitor is orally administered at a dose of about 0.2 mg / kg body weight / week.

[0010] In some embodiments, the cytochrome p450 inhibitor is amiodarone, chloroquine, cimetidine, clomipramine, diphenhydramine, fluoxetine, fluphenazine, haloperidol, paroxetine, perphenazine, propafenone, propoxyphene, quinacrine, quinidine, sertraline, terbinafine, thioridazine, amiodarone, amprenavir, clarithromycin, danazol, delavirdine, diltiazem, efavirenz, erythromycin, ethinyl estradiol, fluconazole, fluvoxamine, grapefruit juice, indinavir, itraconazole, ketoconazole, nefazodone, nelfinavir, quinine, ritonavir, saquinavir, sinacid, troleandomycin, verapamil, or zileuton. In some embodiments, the cytochrome p450 inhibitor is ritonavir.

[0011] In some embodiments, the cytochrome p450 inhibitor is administered at a dose of about 1 mg / kg / week.

[0012] In certain embodiments, the present disclosure provides a method of treating an autoimmune, alloimmune, inflammatory, or mitochondrial condition, the method comprising identifying a subject suffering from one or more of such conditions and administering to the subject a calcineurin inhibitor or a pharmaceutically acceptable salt thereof and a cytochrome p450 inhibitor or a pharmaceutically acceptable salt thereof once a week.

DETAILED DESCRIPTION OF THE INVENTION

[0013] Calcineurin inhibitor Calcineurin inhibitors are widely used in autoimmune, alloimmune, and inflammatory conditions. Cyclosporine, for example, is used to prevent transplant organ rejection (an alloimmune condition) and to treat chronic idiopathic urticaria (an inflammatory condition). Without being bound by theory, it is understood that the mechanism by which calcineurin inhibitors can treat such conditions is to bind to the cytosolic protein cyclophilin in lymphocytes, thereby inhibiting calcineurin in the calcineurin phosphatase pathway. This results in a decrease in the activity of T cells (an important type of white blood cell). This can also have a down-regulatory effect on the entire immune system [Reynolds et al.].

[0014] Calcineurin inhibitors are also used to treat conditions associated with mitochondrial dysfunction. Cyclosporine, for example, has been used to treat muscular dystrophy, a condition associated with mitochondrial dysfunction [Hicks et al.]. Without being bound by theory, it is understood that the mechanism by which calcineurin inhibitors treat mitochondrial conditions is to inhibit MPTP and improve mitochondrial viability [Halestrap et al.].

[0015] Unfortunately, high levels of calcineurin inhibitors can also cause harmful effects. For example, it has been shown that in severe ulcerative colitis, when the blood concentration of cyclosporine exceeds 250 ng / mL, harmful effects including hypertension and nephrotoxicity can occur over the long term [Pham et al.]. On the other hand, it can be difficult to maintain this because the blood concentration of cyclosporine rises rapidly in the first 2 hours after administration and then decreases rapidly 4 hours after administration [Gomez et al.]. Similar results are seen with other calcineurin inhibitors.

[0016] It has previously been shown that when a calcineurin inhibitor (e.g., cyclosporine) is sometimes administered after a cytochrome p450 inhibitor (e.g., itraconazole), the dosing schedule of cyclosporine can be extended by up to 4 hours. However, it has been stated that the only way to extend the dosing schedule of cyclosporine to a once-daily schedule is by co-administering the calcineurin inhibitor and the cytochrome p450 inhibitor with cola [Wimberley et al.].

[0017] Also, it has previously been stated that the only way to improve the linearity of the pharmacokinetics of calcineurin inhibitors is by changing the formulation of the calcineurin inhibitor (such as the microemulsion formulation of cyclosporine known as Neoral) [Mueller et al.].

[0018] This disclosure includes the insight that when a cytochrome p450 inhibitor is co-administered simultaneously or almost simultaneously with a calcineurin inhibitor, the dramatic concentration decrease over the first 4 hours after administration is suppressed and the maintenance of the calcineurin concentration in the bloodstream can be improved. Since calcineurin inhibitors are metabolized by cytochrome p450, inhibiting cytochrome p450 in combination with the active drug extends the half-life of such drugs.

[0019] The present disclosure also encompasses the insight that co - administering a cytochrome p450 inhibitor simultaneously or almost simultaneously with a calcineurin inhibitor would enable dosing frequencies that improve patient compliance, such as once a week, twice a week, etc.

[0020] The methods and compositions claimed herein alleviate various inflammatory, alloimmune, autoimmune, and / or mitochondrial conditions.

[0021] The present disclosure encompasses the insight that by combining inhibition of calcineurin and inhibition of cytochrome p450, inflammation, alloimmunity, and autoimmunity (associated with high lymphocyte levels in some embodiments) can be alleviated. The present disclosure also encompasses the insight that by combining inhibition of calcineurin and inhibition of cytochrome p450, mitochondrial dysfunction can be alleviated.

[0022] Composition In certain embodiments, the methods described herein include the manufacture and use of pharmaceutical compositions and agents comprising compounds identified as active ingredients by the methods described herein. The pharmaceutical compositions themselves are also included.

[0023] In one or more embodiments, the calcineurin inhibitor is selected from the group consisting of cyclosporine, tacrolimus, and pimecrolimus, and analogs or derivatives thereof. In some embodiments, the calcineurin inhibitor is cyclosporine.

[0024] In one or more embodiments, the cytochrome p450 inhibitor is amiodarone, chloroquine, cimetidine, clomipramine, diphenhydramine, fluoxetine, fluphenazine, haloperidol, paroxetine, perphenazine, propafenone, propoxyphene, quinacrine, quinidine, ritonavir, sertraline, terbinafine, thioridazine, amiodarone, amprenavir, clarithromycin, danazol, delavirdine, diltiazem, efavirenz, erythromycin, ethinyl estradiol, fluconazole, fluvoxamine, grapefruit juice, indinavir, itraconazole, ketoconazole, nefazodone, nelfinavir, quinine, ritonavir, saquinavir, sinacide, troleandomycin, verapamil, or zafirlukast. In some embodiments, the cytochrome p450 inhibitor is ritonavir.

[0025] In some embodiments, the compositions disclosed herein include other compounds, drugs, and / or agents used in the treatment of allogeneic immune, autoimmune, and inflammatory conditions. For example, in some cases, the compositions disclosed herein can be combined with one or more (e.g., one, two, three, four, five, or less than ten) compounds.

[0026] In some cases, the compositions disclosed herein are formulated for use as a pharmaceutical composition or for use in a pharmaceutical composition. Such compositions are formulated for or suitable for administration to a subject via any route (e.g., any route approved by the Food and Drug Administration (FDA)). Examples of methods are described in version number 004 of the FDA's CDER Data Standards Manual (available at fda.give / cder / dsm / DRG / drg00301.htm). The pharmaceutical compositions described herein can be formulated for oral delivery, parenteral delivery, or transdermal delivery. The compounds of the present disclosure can also be combined with other pharmaceuticals.

[0027] In some aspects, the present disclosure provides a kit comprising one or more compositions comprising cyclosporine and / or ritonavir and / or itraconazole (in separate compositions or in a single composition). The kit may also include instructions for physicians and / or patients, syringes, needles, boxes, bottles, vials, and the like.

[0028] In some cases, the methods described herein involve administering an effective amount of a composition(s) comprising a calcineurin inhibitor and a cytochrome p450 inhibitor (as part of a single composition or as separate compositions) as described above. As used herein, the terms "effective amount" and "therapeutically effective" refer to the amount or concentration of one or more drugs that is effective for a period of time (including acute or chronic administration and intermittent or continuous administration) in bringing about a desired effect or physiological outcome when administered in the context of administering one or more drugs.

[0029] In some cases, the composition comprises a calcineurin inhibitor (e.g., cyclosporine), a cytochrome p450 inhibitor (e.g., ritonavir), and a pharmaceutically acceptable carrier, adjuvant, and / or vehicle. In some cases, the compositions described herein further comprise one or more additional therapeutic agents in an amount effective to achieve modulation of a disease or disease symptom.

[0030] The term "pharmaceutically acceptable carrier or adjuvant" refers to a carrier or adjuvant that can be administered to a patient together with a compound of the present disclosure in a dosage sufficient to effect delivery of a therapeutically effective amount of the compound without impairing the pharmacological activity of the compound and that is non-toxic. As used herein, the term "pharmaceutically acceptable carrier" includes saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are compatible with pharmaceutical administration.

[0031] The composition can typically be formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral administration (e.g., intravenous administration, intradermal administration, subcutaneous administration), oral administration (e.g., inhalation administration), transdermal administration (topical administration), transmucosal administration, and rectal administration.

[0032] The composition can be in the form of a solution or powder for inhalation and / or nasal administration. Such compositions can be formulated according to techniques known in the art using suitable dispersing or wetting agents (e.g., Tween® 80, etc.) and suspending agents. A sterile preparation for injection can also be a sterile solution or suspension for injection in a parenterally acceptable non-toxic diluent or solvent (e.g., a solution in 1,3 - butanediol). Acceptable media and solvents that can be used include mannitol, water, Ringer's solution, and isotonic sodium chloride solution. Further, a sterile non-volatile oil is usually used as the solvent or suspending medium. For this purpose, any non-irritating non-volatile oil containing synthetic monoglycerides or diglycerides can be used. Fatty acids (such as oleic acid and its glyceride derivatives) are useful in the preparation of injectables because they are pharmaceutically acceptable natural oils (such as olive oil or castor oil, especially their polyoxyethylated versions). Such oily solutions or suspensions can also include long-chain alcohol diluents or dispersing agents, or carboxymethyl cellulose, or similar dispersing agents generally used in the formulation of pharmaceutically acceptable dosage forms (such as emulsions and / or suspensions). Other commonly used surfactants (such as Tween or Span®) and / or other similar emulsifying agents or bioavailability enhancing agents generally used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms can also be used for the purpose of formulation.

[0033] The composition can be administered orally in any orally acceptable dosage form, including but not limited to capsules, tablets, emulsions, and aqueous suspensions, dispersions, and solutions. For tablets for oral use, commonly used carriers include lactose and corn starch. Lubricants (such as magnesium stearate) are also typically added. For oral administration in capsule form, useful diluents include lactose and dried corn starch. When an aqueous suspension and / or emulsion is administered orally, the active ingredient can be suspended or dissolved in an oily phase in combination with an emulsifying agent and / or suspending agent. If desired, certain sweetening agents and / or flavoring agents and / or coloring agents can be added.

[0034] Alternatively or additionally, the pharmaceutical composition can be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the pharmaceutical art and can be prepared as solutions in physiological saline using benzyl alcohol or other suitable preservatives, absorption promoters for enhancing bioavailability, fluorocarbons, and / or other solubilizing or dispersing agents known in the art.

[0035] In some embodiments, the present disclosure provides a method for administering a composition comprising a calcineurin inhibitor (e.g., cyclosporine) and a composition comprising a cytochrome p450 inhibitor (e.g., ritonavir), each comprising a pharmaceutical composition disclosed herein (hereinafter referred to as "X") in the following method: a substance X for use as an agent in the treatment of one or more of the diseases or conditions disclosed herein (e.g., inflammation, referred to as "Y" in the examples below). Use of substance X for the manufacture of an agent for the treatment of Y, and use of substance X for use in the treatment of Y.

[0036] Optionally, the therapeutic compositions disclosed herein can be formulated for sale in the United States, import into the United States, and / or export from the United States.

[0037] The pharmaceutical composition can be included in a container, pack, or dispenser, together with instructions for administration.

[0038] Dosage In some embodiments, a method of treating an autoimmune, alloimmune, or inflammatory condition comprises administering a calcineurin inhibitor and a cytochrome p450 inhibitor.

[0039] In some embodiments, the calcineurin inhibitor and the cytochrome p450 inhibitor are administered simultaneously.

[0040] In some embodiments, the calcineurin inhibitor and the cytochrome p450 inhibitor are administered sequentially.

[0041] In some aspects of the present disclosure, the calcineurin inhibitor and the cytochrome p450 inhibitor are administered separately (i.e., in separate dosage forms).

[0042] In some embodiments, the calcineurin inhibitor is administered in an amount of about 0.1 mg / kg body weight / week, about 0.5 mg / kg body weight / week, about 1 mg / kg body weight / week, about 2 mg / kg body weight / week, or about 4 mg / kg body weight / week. In some embodiments, the cytochrome p450 inhibitor is administered in an amount of 1 mg / kg / week, 2 mg / kg / week, 5 mg / kg / week, or 10 mg / kg / week. In some embodiments, the cytochrome p450 inhibitor is administered in an amount of 5 mg / week, 10 mg / week, or 20 mg / week. These compounds can be administered separately or together, including as part of a treatment regimen.

[0043] In some aspects of the present disclosure, the calcineurin inhibitor is cyclosporine and the cytochrome p450 inhibitor is ritonavir. In some aspects, cyclosporine and ritonavir are administered separately (i.e., in separate dosage forms). In some embodiments, cyclosporine is administered in an amount of 0.1 mg / kg body weight / week, about 0.5 mg / kg body weight / week, about 1 mg / kg body weight / week, about 2 mg / kg body weight / week, or about 4 mg / kg body weight / week. In some embodiments, ritonavir is administered in an amount of 1 mg / kg / week, 2 mg / kg / week, 5 mg / kg / week, or 10 mg / kg / week. In some embodiments, ritonavir is administered in an amount of 5 mg / week, 10 mg / week, or 20 mg / week. The compounds described herein can be administered separately or together, including as part of a treatment regimen.

[0044] In some embodiments, cyclosporine is administered in an amount of about 1.0 - 2.5 mg / kg. In some embodiments, cyclosporine is administered in an amount of about 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, or 2.5 mg / kg. In some embodiments, cyclosporine is administered in an amount of about 5 - 15 mg. In some embodiments, cyclosporine is administered in an amount of about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 29 mg. In some embodiments, ritonavir is administered in an amount of 5 mg, 10 mg, or 20 mg. These compounds can be administered separately or together, including as part of a treatment regimen. In some embodiments, a single dose of cyclosporine and ritonavir is administered, followed by a different dose after a certain period (e.g., 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 1 week).

[0045] In some embodiments, the subject may be administered a specific dose over several days, weeks, months, or years. For example, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, etc. In some embodiments, the subject is administered a calcineurin inhibitor and a cytochrome inhibitor according to an intermittent dosing schedule (e.g., a frequency of less than once a day, e.g., once every other day, twice a week, once a week, twice a month, once a month, twice a year, once a year, etc.). In some embodiments, the subject is administered a calcineurin inhibitor and a cytochrome inhibitor every other day. In some embodiments, the subject is administered a calcineurin inhibitor and a cytochrome inhibitor twice a week. In some embodiments, the subject is administered a calcineurin inhibitor and a cytochrome inhibitor once a week. In some embodiments, the subject is administered a calcineurin inhibitor and a cytochrome inhibitor once every other week. In some embodiments, the subject is administered a calcineurin inhibitor and a cytochrome inhibitor twice a month. In some embodiments, the subject is administered a calcineurin inhibitor and a cytochrome inhibitor once a month. In some embodiments, the subject is administered a calcineurin inhibitor and a cytochrome inhibitor once every other month. In some embodiments, the subject is administered a calcineurin inhibitor and a cytochrome inhibitor twice a year. In some embodiments, the subject is administered a calcineurin inhibitor and a cytochrome inhibitor once a year.

[0046] In some embodiments, the calcineurin inhibitor is cyclosporine and the cytochrome p450 inhibitor is ritonavir. In some embodiments, cyclosporine and ritonavir or itraconazole are administered separately or together as a single daily dose, on a daily basis, on a weekly basis, or in some other unit, for several weeks, several months, or several years. For example, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, etc.

[0047] Treatment method The methods described herein include methods for treating disorders associated with elevated lymphocyte levels (e.g., transplant organ rejection) or mitochondrial dysfunction (e.g., muscular dystrophy). Generally, the method involves combining a therapeutically effective amount of a calcineurin inhibitor (e.g., cyclosporine) with a cytochrome p450 inhibitor (e.g., itraconazole) as described herein and administering it to a subject in need of such treatment or a subject identified as being in need of such treatment (e.g., a mammalian subject, e.g., a human subject).

[0048] In some cases, the method may include selecting a human subject who has or had a condition or disease. In some cases, suitable subjects include, for example, subjects who have or had a condition or disease, but the disease or an aspect thereof has resolved, the symptoms of the disease have been reduced (e.g., compared to other subjects with the same condition or disease (e.g., most subjects)), and / or the period of survival with the condition or disease (e.g., compared to other subjects with the same condition or disease (e.g., most subjects)) has been extended (e.g., asymptomatic).

[0049] The methods disclosed herein can be applied to a wide range of species (e.g., humans, non-human primates (e.g., monkeys), horses, cows, pigs, sheep, deer, elk, goats, dogs, cats, rabbits, guinea pigs, hamsters, rats, and mice).

[0050] The terms "treat," "treating," "treatment," etc., as applied to isolated cells, include subjecting the cells to any type of process or condition, or performing any type of operation or procedure on the cells. The term "treating" as applied to a subject refers to providing medical or surgical care, nursing, or management to an individual. An individual is typically one who has a disease or injury, or has an increased risk of getting a disease compared to the average member of a population, and who requires such care, nursing, or management.

[0051] In some embodiments, the terms "treating" and "treatment" refer to administering to a subject an effective amount of a composition (e.g., a composition comprising a calcineurin inhibitor and a composition comprising a cytochrome p450 inhibitor), as a result of which there is a reduction in at least one symptom of a disease or an improvement in the disease (e.g., a favorable clinical outcome or a desired clinical outcome) in the subject. For the purposes of the present disclosure, favorable clinical outcomes or desired clinical outcomes include, but are not limited to, alleviation of one or more symptoms, whether detectable or undetectable, reduction in the degree of the disease, stabilization of the disease state (i.e., no worsening), delay or blunting of the progression of the disease, amelioration or reduction of the disease state, and remission (partial or complete). Treating can refer to extending the survival period compared to the expected survival period if treatment were not given. Thus, one of ordinary skill in the art will recognize that treatment can improve the disease state but may not result in a cure of the disease. In some embodiments, treatment can be prophylactic treatment, in which a composition disclosed herein is administered to a subject at risk of developing an inflammation disclosed herein. In some embodiments, treatment is "effective" when the progression of the disease is blunted or stopped.

[0052] As used herein, the term "subject" refers to any animal. In some cases, the subject is a mammal. In some cases, the term "subject" as used herein refers to a human (e.g., male, female, or pediatric). In some embodiments, the "subject" is a non-human mammal, such as a non-human primate (e.g., monkey), horse, cow, pig, sheep, deer, elk, goat, dog, cat, rabbit, guinea pig, hamster, rat, and mouse.

[0053] In some cases, subject selection may include obtaining a sample from a subject (e.g., a candidate subject) and examining the sample for an indicator suitable for subject selection. In some cases, the subject may have had or have a condition or disease that can be confirmed or identified (e.g., by a healthcare provider). In some cases, showing a positive immune response to a condition or disease can be identified from patient records, family history, and / or detection of an indicator of a positive immune response. In some cases, multiple parties may be involved in subject selection. For example, a first party may obtain a sample from a candidate subject, and a second party may examine the sample. In some cases, the subject may be selected and / or designated by a physician (e.g., a general practitioner). In some cases, subject selection may include obtaining a sample from the selected subject, storing the sample, and / or using the methods disclosed herein. The sample may include, for example, cells or a cell population.

[0054] In some cases, the treatment method may include single-dose, multiple-dose, and repeated-dose administration depending on the prevention or treatment of the disease or condition that the subject is suffering from. In some cases, the treatment method may include evaluating the level of the disease in the subject before, during, and / or after treatment. In some cases, the treatment may be continued until a decrease in the level of the disease in the subject is detected.

[0055] As used herein, the terms "administer", "administering", or "administration" refer to implanting, absorbing, ingesting, injecting, or inhaling the drug of the present invention, regardless of form. In some cases, one or more of the compounds disclosed herein may be administered to a subject locally (e.g., nasally) and / or orally. For example, the methods described herein include administering an effective amount of a compound or compound composition to achieve the desired or described effect. The specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, weight, general health, sex, diet, time of administration, rate of excretion, drug combinations, disease, condition, or severity and course of symptoms, susceptibility of the patient to the disease, condition, or symptom, and the judgment of the physician treating the patient.

[0056] After administration, the subject can be examined so that the level of the disease is detected, evaluated, or determined. In some cases, treatment can be continued until a change (e.g., decrease) in the level of the disease in the subject is detected.

[0057] When an improvement in the patient's condition (e.g., a change (e.g., decrease) in the level of the disease in the subject) occurs, the compounds, compositions, or combinations of the present disclosure can be administered at a maintenance dose, if necessary. Thereafter, the dosage or frequency of administration or both can be reduced according to the symptoms to a level at which the improvement in the condition is maintained. On the other hand, intermittent treatment may be required for the patient in the long term upon recurrence of any of the disease symptoms.

[0058] Definitions Neurodegeneration refers to any condition in which nerve cells die progressively.

[0059] A mitochondrial condition refers to any condition resulting from mitochondrial dysfunction.

[0060] Inhibitory agent: As used herein, the term "inhibitory agent" refers to an entity, condition, or event whose presence, level, or degree correlates with a decrease in the level or activity of a target. In some embodiments, the inhibitory agent can act directly (in which case the inhibitory agent exerts its effect directly on its target, e.g., by binding to the target). In some embodiments, the inhibitory agent can act indirectly (in which case the inhibitory agent exerts its effect by interacting with a regulator of the target and / or changing the regulator of the target in some other manner such that the level and / or activity of the target is decreased). In some embodiments, the inhibitory agent is one whose presence or level correlates with a decrease in the target level or target activity as compared to a particular reference level or reference activity (e.g., that observed under appropriate reference conditions such as the presence of a known inhibitory agent or the absence of the inhibitory agent in question).

[0061] As used herein, inhibitor refers to an inhibitory agent, while inhibition refers to the activity of the inhibitor agent.

[0062] Controlling: Controlling refers to changing, enhancing, or decreasing the activity or an organelle or cell.

[0063] Antagonist: Those skilled in the art will understand that the term "antagonist" as used herein can be used to refer to an agent, condition, or event whose presence, level, degree, type, or form correlates with a decrease in the level or activity of another substance (i.e., the substance or target being inhibited). Generally, an antagonist can be, or can include, an agent of any chemical class, such agents including, for example, small molecules, polypeptides, nucleic acids, carbohydrates, lipids, metals, and / or any other entity exhibiting related inhibitory activity. In some embodiments, the antagonist can be direct (in which case the antagonist exerts its effect directly on its target). In some embodiments, the antagonist can be indirect (in which case the antagonist exerts its effect by something other than binding to its target (e.g., by interacting with a regulator of the target such that the level or activity of the target is altered)).

[0064] Agonist: Those skilled in the art will understand that the term "agonist" as used herein can be used to refer to an agent, condition, or event whose presence, level, degree, type, or form correlates with an increase in the level or activity of another substance (i.e., the substance or target being agonized). Generally, an agonist can be, or can include, an agent of any chemical class, such agents including, for example, small molecules, polypeptides, nucleic acids, carbohydrates, lipids, metals, and / or any other entity exhibiting related activating activity. In some embodiments, the agonist can be direct (in which case the agonist exerts its effect directly on its target). In some embodiments, the agonist can be indirect (in which case the agonist exerts its effect by something other than binding to its target (e.g., by interacting with a regulator of the target such that the level or activity of the target is altered)).

[0065] Administration: As used herein, the term "administration" typically refers to the application or delivery to a subject or system. Those skilled in the art, upon reading this disclosure, will understand that various routes are available for the administration of, for example, a composition. For example, depending on the composition, it can be administered by one or more routes (such as ophthalmic, oral, parenteral, topical, etc.). In some specific embodiments, the administration can be to the bronchus (e.g., by intratracheal instillation), oral cavity, skin (e.g., one or more of or including intradermal topical administration, intradermal topical administration, intradermal topical administration, transdermal topical administration, etc.), enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, subarachnoid, intravenous, intraventricular, within a specific organ (e.g., intrahepatic), mucosa, transnasal, oral, rectal, subcutaneous, sublingual, topical, trachea (e.g., by intratracheal instillation), vagina, vitreous body, etc. Further, in some embodiments, this disclosure describes administering behavioral therapy, in some embodiments, for example, via interaction with a counselor (e.g., a therapist) as described herein and / or using a device or computer system. In some embodiments, the administration includes intermittent dosing, application, or interaction (e.g., multiple doses separated in time, for example, in some embodiments, a frequency of less than once a day, for example, once every other day, twice a week, once a week, twice a month, once a month, twice a year, once a year, etc.). In some embodiments, the administration of a calcineurin inhibitor and a cytochrome p450 inhibitor to a subject follows an intermittent dosing schedule. In some embodiments, the administration of a calcineurin inhibitor and a cytochrome p450 inhibitor to a subject is performed at a frequency of less than once a day. In some embodiments, the calcineurin inhibitor and the cytochrome p450 inhibitor are each administered to the subject once a week. In some embodiments, the calcineurin inhibitor and the cytochrome p450 inhibitor are each administered to the subject twice a week. In some embodiments, the calcineurin inhibitor and the cytochrome p450 inhibitor are each administered to the subject once a month. In some embodiments, the calcineurin inhibitor and the cytochrome p450 inhibitor are each administered to the subject twice a month.In some embodiments, the calcineurin inhibitor and the cytochrome p450 inhibitor are each administered to the subject twice a year. In some embodiments, the calcineurin inhibitor and the cytochrome p450 inhibitor are each administered to the subject once a year.

[0066] Cyclosporine is a compound having the following structure.

Chemical formula

[0067] Ritonavir is a compound having the following structure.

Chemical formula

[0068] About: As used herein with respect to a value, the term "about" refers to a value being similar in the context of the reference value. In general, one of ordinary skill in the art will be familiar with the context and will understand the appropriate degree of variation subsumed by "about" in that context. For example, in some embodiments, the term "about" may encompass a range of values within 25%, within 20%, within 19%, within 18%, within 17%, within 16%, within 15%, within 14%, within 13%, within 12%, within 11%, within 10%, within 9%, within 8%, within 7%, within 6%, within 5%, within 4%, within 3%, within 2%, within 1%, or less than the reference value.

[0069] Pharmaceutically acceptable salts: The term "pharmaceutically acceptable salt(s)" refers to salts formed from acidic and basic groups of pharmaceutically active compounds. Examples of salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, metaphosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)).

[0070] Unless otherwise defined, 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 disclosure belongs. Methods and materials are described herein for use in this disclosure, and in addition, appropriate methods and materials known to those skilled in the art can be used. The materials, methods, and examples are illustrative only and not intended to be limiting. Publications, patent applications, patents, sequences, database entries, and other references mentioned herein are all incorporated by reference in their entirety. In case of conflict, this specification, including definitions, will control.

[0071] Other features and advantages of the present disclosure will become apparent from the following detailed description, figures, and claims.

[0072] The present disclosure is further illustrated in the following examples, which do not limit the scope of the present disclosure as claimed.

Examples

[0073] The present disclosure is further illustrated in the following examples, which do not limit the scope of the present disclosure described in the claims.

[0074] Example 1 Materials and Methods Test the compounds in the escalating dose study described herein.

[0075] Obtain modified cyclosporine (Neoral) and ritonavir.

[0076] Screen 18 cats for participation in the escalating dose study. The inclusion criteria are over 6 months of age and weighing 3.5 - 6 kg. The exclusion criteria are the presence of any clinically significant acute or chronic disease (in the opinion of the study investigator) after a detailed medical and surgical history and a complete physical examination.

[0077] After obtaining permission to participate, divide the subjects into 3 cohorts: Cohort 1, Cohort 2, and Cohort 3. Each subject participates in only one cohort. Each cohort will include 6 cats.

[0078] In Cohort 1, administer 1.2 mg of oral cyclosporine and a single oral dose of 5 mg of ritonavir simultaneously.

[0079] In Cohort 2, administer 1.2 mg of oral cyclosporine and a single oral dose of 10 mg of ritonavir simultaneously.

[0080] In Cohort 3, administer 1.2 mg of oral cyclosporine and a single oral dose of 20 mg of ritonavir simultaneously.

[0081] Collect samples from each cohort for 7 days from the oral dose of cyclosporine and the oral doses of cyclosporine and ritonavir administered simultaneously.

[0082] Results Determine the whole blood cyclosporine concentration for each sample. Also, for each sample, estimate the whole blood PK parameters using non-compartmental analysis as needed: Cmax, tmax, kel, t1 / 2, AUC0-last, AUC0-inf, CL / F, and Vz / F. Also, measure the plasma ritonavir concentration to confirm its presence after dosing on day 4.

[0083] Calculate descriptive statistics for all relevant PK parameters: n, mean, standard deviation, minimum, median, maximum, geometric mean, and coefficient of variation.

[0084] Use the natural logarithm of the dose-uncorrected parameters and an analysis of variance (ANOVA) model with the subject as a random effect and the day as a fixed effect to compare the PK parameters (Cmax, AUC0-last, AUC0-inf) between day 1 and day 4. Construct a confidence interval (CI) (90%) for the geometric mean ratio (GMR) of cyclosporine from day 4 to day 1 for all three parameters using the log-transformed data and a two one-sided t-test procedure. Reverse the GMR and 90% CI to the original scale. Evaluate the effect of co-administration of ritonavir with cyclosporine from the GMR and CI.

[0085] This analysis will show a statistically significant effect on the extension of the cyclosporine half-life suitable for a once-weekly dosing regimen.

[0086] Example 2 Materials and Methods The compounds were tested in the comparative dosing study described herein.

[0087] Modified cyclosporine (Atopica) and ritonavir were obtained.

[0088] Fourteen cats were screened for participation in the escalating dose study. Of these, twelve were selected. The inclusion criteria were over 6 months of age and weighing 3.5 - 6 kg. The exclusion criteria were the presence of any clinically significant evidence of acute or chronic disease (in the opinion of the study investigator) after a detailed medical and surgical history and a complete physical examination, and any other factors that, in the opinion of the study investigator, might affect the purpose of the study, such as recent medical treatment.

[0089] After obtaining permission to participate, the subjects were acclimated to the study conditions for 7 days prior to the start of the study. The subjects were administered 10 mg of modified cyclosporine in Torpac capsules, and their condition was monitored for 7 days.

[0090] Throughout the study, physical measurements and clinical observations were performed. Blood samples were also collected once before the 7-day period after administration, once after the study period, and 15 times throughout the 7-day period. The blood samples were analyzed.

[0091] After a 10-day washout period, Period 2 was initiated. The subjects were administered 10 mg of cyclosporine (Atopica) in Torpac capsules and 20 mg of ritonavir solution in Torpac capsules. The same procedures were performed for physical measurements, clinical observations, and blood samples.

[0092] Results The whole blood cyclosporine concentration was determined for each sample. For each sample, non-compartmental analysis was used to estimate the whole blood PK parameters as needed: Cmax, tmax, kel, t1 / 2, AUC0-last, AUC0-inf, CL / F, and Vz / F. Plasma ritonavir concentration was also measured at 11 time points within the first 24 hours during Period 2.

[0093] Descriptive statistics were calculated for all relevant PK parameters: n, mean, standard deviation, minimum, median, maximum, geometric mean, and coefficient of variation.

[0094] Use an analysis of variance (ANOVA) model with the variables being the effect and the number of days to compare the PK parameters (Cmax, AUC0 - last, AUC0 - inf) between Day 1 and Day 4.

[0095] The following table provides an overview of the pharmacokinetic parameters of cyclosporine after oral administration of 10 mg to cats, with and without ritonavir (20 mg). Due to dose variability, two cats were excluded from the study.

Table 1

Claims

1. 1. A method for treating an alloimmune, autoimmune, inflammatory, or mitochondrial condition in a subject, the method comprising administering to the subject a calcineurin inhibitor and a cytochrome p450 inhibitor according to an intermittent dosing schedule.

2. 2. The method of claim 1, wherein the calcineurin inhibitor and the cytochrome p450 inhibitor are administered once a week.

3. The method of claim 1 , wherein the calcineurin inhibitor and the cytochrome p450 inhibitor are administered simultaneously.

4. The method of claim 1 , wherein the calcineurin inhibitor and the cytochrome p450 inhibitor are administered sequentially.

5. The method of any one of claims 1 to 3, wherein the calcineurin inhibitor is selected from cyclosporine, tacrolimus, pimecrolimus, and analogues or derivatives thereof, or pharma- ceutically acceptable salts thereof.

6. The cytochrome p450 inhibitor is amiodarone, chloroquine, cimetidine, clomipramine, diphenhydramine, fluoxetine, fluphenazine, haloperidol, paroxetine, perphenazine, propafenone, propoxyphene, quinacrine, quinidine, ritonavir, sertraline, terbinafine, thioridazine, amiodarone, amprenavir, clarithromycin, danazol, delavirdine, diltiazem, efavirenz , erythromycin, ethinyl estradiol, fluconazole, fluvoxamine, grapefruit juice, indinavir, itraconazole, ketoconazole, nefazodone, nelfinavir, quinine, ritonavir, saquinavir, synacid, troleandomycin, verapamil, zafirlukast, and analogs or derivatives thereof, or pharma- ceutically acceptable salts thereof.

7. 2. The method of claim 1, wherein the calcineurin inhibitor is cyclosporine.

8. 2. The method of claim 1, wherein the cytochrome p450 inhibitor is ritonavir.

9. The method according to any one of claims 1 to 7, wherein the calcineurin inhibitor is administered in an amount of about 0.1 mg / kg body weight / week, about 0.2 mg / kg body weight / week, about 0.3 mg / kg body weight / week, about 0.4 mg / kg body weight / week, about 0.5 mg / kg body weight / week, about 1.0 mg / kg body weight / week, about 2.0 mg / kg body weight / week, or about 4.0 mg / kg body weight / week.

10. The method according to any one of claims 1 to 8, wherein the cytochrome p450 inhibitor is administered in an amount of about 0.1 mg / kg body weight / week, about 0.2 mg / kg body weight / week, about 0.3 mg / kg body weight / week, about 0.4 mg / kg body weight / week, about 0.5 mg / kg body weight / week, about 1.0 mg / kg body weight / week, about 2.0 mg / kg body weight / week, or about 4.0 mg / kg body weight / week.

11. The method according to any one of claims 1 to 8, wherein the cytochrome p450 inhibitor is administered in an amount of about 1 mg / week, about 2 mg / week, about 5 mg / week, about 10 mg / week, about 20 mg / week, or about 40 mg / week.

12. The method according to any one of claims 1 to 11, wherein the autoimmune, alloimmune, or inflammatory condition is associated with an elevated level of lymphocytes.

13. The method according to any one of claims 1 to 11, wherein the mitochondrial condition is associated with the mitochondrial permeability transition pore.

14. The method according to claim 12, wherein the autoimmune, alloimmune, or inflammatory condition is atopy, perianal fistula, canine exfoliative pemphigus, sebaceous adenitis, granulomatous meningoencephalitis, inflammatory bowel disease, or stomatitis.

15. The method according to any one of claims 1 to 14, wherein the calcineurin inhibitor is administered in an oral dosage form.

16. The method according to any one of claims 1 to 15, wherein the cytochrome p450 inhibitor is administered in an oral dosage form.

17. A method of intervening in a disease, disorder, or condition associated with an elevated level of lymphocytes in a subject, the method comprising administering a calcineurin inhibitor and a cytochrome p450 inhibitor to the subject according to an intermittent dosing schedule.

18. A method of intervening in a disease, disorder, or condition associated with the mitochondrial permeability transition pore in a subject, the method comprising administering a calcineurin inhibitor and a cytochrome p450 inhibitor to the subject according to an intermittent dosing schedule.

19. The method according to claim 17 or 18, wherein the calcineurin inhibitor and the cytochrome p450 inhibitor are administered simultaneously.

20. The method according to any one of claims 17 to 19, wherein the calcineurin inhibitor and the cytochrome p450 inhibitor are administered continuously.

21. The method according to any one of claims 17 to 20, wherein the calcineurin inhibitor is selected from cyclosporine, tacrolimus, and pimecrolimus, and their analogs or derivatives, or pharmaceutically acceptable salts thereof.

22. The method according to any one of claims 17 to 21, wherein the cytochrome p450 inhibitor is selected from amiodarone, chloroquine, cimetidine, clomipramine, diphenhydramine, fluoxetine, fluphenazine, haloperidol, paroxetine, perphenazine, propafenone, propoxyphene, quinacrine, quinidine, ritonavir, sertraline, terbinafine, thioridazine, amiodarone, amprenavir, clarithromycin, danazol, delavirdine, diltiazem, efavirenz, erythromycin, ethinyl estradiol, fluconazole, fluvoxamine, grapefruit juice, indinavir, itraconazole, ketoconazole, nefazodone, nelfinavir, quinine, ritonavir, saquinavir, sinacid, troleandomycin, verapamil, or zileuton, and their analogs or derivatives, or pharmaceutically acceptable salts thereof.

23. The method according to any one of claims 17 to 22, wherein the calcineurin inhibitor and the cytochrome p450 inhibitor are administered once a week.

24. The method according to any one of claims 1 to 23, wherein the subject is a non-human mammal.

25. The method according to claim 24, wherein the subject is a cat or a dog.

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