Purine Compounds for Treating Disorders - Patent application

Abstract

The substituted purine compounds of formulas (I)-(III) and their salts act as adenosine A2a receptor (A2aR) antagonists for cancer immunotherapy, depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's disease, and Parkinson's disease. Pharmaceutical compositions containing such compounds and methods for their use in treating depression are also taught.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates generally to substituted purine compounds and salts thereof that act as adenosine A2a receptor (A2aR) antagonists for the treatment of cancer, depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's, and Parkinson's, pharmaceutical compositions containing such compounds, and methods of their use in treating cancer, depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's, and Parkinson's. [Background technology]

[0002] Background of the Invention Adenosine exerts its biological effects through a class of membrane-specific receptors that belong to the superfamily of G protein-linked receptors. At least four subtypes of adenosine receptors have been identified: A1, A2a, A1b, and A3.

[0003] A2aR has been shown to play a regulatory role in the immune system. One A2aR antagonist, istradefylline, has been shown to reduce motor dysfunction, which in turn improves function in neurodegenerative diseases such as Parkinson's disease and related movement disorders (e.g., Huntington's disease).

[0004] WO2013058681A2 discloses the use of A2aR antagonists to treat diseases of the central nervous system, tumor diseases, and viral and bacterial diseases. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] WO2013058681A2 [Patent Document 2] U.S. Patent No. 4,938,949 [Non-patent literature]

[0006] [Non-Patent Document 1] Berge, SM et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 66:1-19, 1977 [Non-patent document 2] Robert D. Leone, Ying-Chun Lo and Jonathan D. Powell, “A2aR antagonists: Next generation checkpoint blockade for cancer immunotherapy”, Comput Struct Biotechnol J. 2015; 13: 265~272 [Non-patent document 3] Domenici MR et al., “Adenosine A2A receptor as potential therapeutic target in neuropsychiatric disorders”, Pharmacol Res 2019; 147: 104338 [Non-patent document 4] Cronstein B. “Adenosine receptors and fibrosis: a translational view”, F1000 Biol Rep. 2011; 3: 21 [Non-patent document 5] The Pharmacological Basis of Therapeutics, edited by Goodman and Gilman, Macmillan Publishing Co., New York Summary of the Invention [Problem to be solved by the invention]

[0007] There is a need for A2aR antagonists suitable for treating neurological disorders, fibrosis-related disorders (NASH and scleroderma) and cancer. [Means for solving the problem]

[0008] SUMMARY OF THE INVENTION The present inventors have discovered certain purine compounds that are useful as A2aR antagonists for treating diseases such as depression.

[0009] In one aspect, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof: [ka] 8-[(E)-2-[3,4-bis(difluoromethoxy)phenyl]vinyl]-1,3-diethyl-7-methyl-purine-2,6-dione; Hydrofluoride

[0010] In one aspect, there is provided a compound of formula (II) or a pharmaceutically acceptable salt thereof: [ka] 8-[(E)-2-[3,4-bis(fluoromethoxy)phenyl]vinyl]-1,3-diethyl-7-methyl-purine-2,6-dione

[0011] In one aspect, there is provided a compound of formula (III) or a pharmaceutically acceptable salt thereof: [ka] 8-[(E)-2-[3,4-dimethoxyphenyl]vinyl]-1,3-diethyl-7-methyl-6-thioxopurin-2-one

[0012] In one embodiment, istradefylline, E)-8-(3,4-dimethoxystyryl)-1,3-diethyl-7-methylxanthine of formula (IV), or a pharmaceutically acceptable salt thereof, is found to be an A2aR antagonist that may be useful for treating cancer, depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's and Parkinson's. [ka]

[0013] In one embodiment, SCH 442416, 2-(2-furanyl)-7-[3-(4-methoxyphenyl)propyl]-7H-pyrazolo[4,3-e][1,2,4]triazolo[1,5-c]pyrimidin-5-amine of formula (V), or a pharmaceutically acceptable salt thereof, is found to be an A2aR antagonist that may be useful for treating cancer, depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's and Parkinson's. [ka]

[0014] The present disclosure also includes pharmaceutical compositions comprising a therapeutically effective amount of one or more of the compounds of Formula I, II and III and pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable excipient.

[0015] The present disclosure further includes methods for treating depression using one or more of the compounds of Formula I, II, and III and pharmaceutically acceptable salts thereof, the methods comprising administering one or more compounds to a subject in need of such treatment, thereby treating depression.

[0016] The present disclosure further includes methods for treating cancer, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's, or Parkinson's using one or more of the compounds of Formula I, II, and III and pharmaceutically acceptable salts thereof. The methods include administering one or more compounds to a subject in need of such treatment, thereby treating depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's, or Parkinson's.

[0017] In some embodiments, the compound is administered by intravenous injection, by injection into a tissue, intraperitoneally, orally, or intranasally. In some embodiments, the composition has the form of a solution, dispersion, suspension, powder, capsule, tablet, pill, sustained release capsule, sustained release tablet, or sustained release pill.

[0018] Methods are provided for synthesizing compounds of Formulas I, II, and III.

[0019] The disclosure includes methods that include providing at least one such compound, measuring the inhibition of A2aR activity for the compound, and determining whether the inhibition is greater than expected. BRIEF DESCRIPTION OF THE DRAWINGS [Brief explanation of the drawings]

[0020] [Figure 1] 1 is a graph showing the results of a GPCR screening of compounds of formula I. [Figure 2] 1 is a graph showing the results of a GPCR screening of compounds of formula II. [Figure 3] 1 is a graph showing the results of a GPCR screening of SD-007, a compound of Formula III. [Figure 4] 1 is a graph showing the results of a GPCR screen of istradefyline, a compound of Formula IV. [Figure 5] 1 is a graph showing the results of a GPCR screening of 5′-N-ethylcarboxamidoadenosine (NECA). [Figure 6] 1 is a graph showing the results of GPCR screening of SCH 442416. [Figure 7] 1 is a graph showing plasma and brain concentrations of the compound of formula I over time. [Figure 8] 1 is a graph showing plasma concentrations of a compound of formula I over time following administration of 1 mg / kg intravenously or 3 mg / kg orally. [Figure 9] 1 is a graph showing plasma and brain concentrations of the compound of formula II over time. [Figure 10] 1 is a graph showing plasma concentrations over time following administration of 1 mg / kg intravenously or 3 mg / kg orally of a compound of Formula II. [Figure 11] 1 is a graph showing plasma and brain concentrations of the compound of formula III, SD-007, over time. [Figure 12] 1 is a graph showing the plasma concentration over time of the compound of formula III, SD-007, following administration of 1 mg / kg intravenously or 3 mg / kg orally. [Figure 13] 1 is a graph showing plasma and brain concentrations of the compound Istradefylline (Formula IV) over time. [Figure 14] 1 is a graph showing plasma concentrations over time following administration of 1 mg / kg intravenously or 3 mg / kg orally of the compound of formula IV. [Figure 15] 1 is a graph comparing latency to immobility times for various compounds. [Figure 16] 1 is a graph comparing immobility time for various compounds. DETAILED DESCRIPTION OF THE INVENTION

[0021] Detailed Description Embodiments of the present disclosure are discussed in detail below. In describing the embodiments, specific terminology is used for clarity. However, the present disclosure is not intended to be limited to the specific terminology so selected. Those skilled in the art will recognize that other equivalents can be used and other methods can be developed without departing from the spirit and scope of the present disclosure. All references cited herein are incorporated by reference as if each were individually incorporated.

[0022] 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.

[0023] As used in the specification and claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0024] The term "comprising," as used herein, will be understood to mean that the list that follows is not exhaustive and may or may not include any other additional suitable items, such as one or more further features, components and / or ingredients, as appropriate.

[0025] The terms "pharmaceutically effective amount," "therapeutically effective amount," or "therapeutically effective dose," "effective amount" refer to an amount of a subject compound that will elicit the biological or medical response in a tissue, system, animal, or human that is desired by a researcher, veterinarian, physician, or other clinician. The term "therapeutically effective amount" includes an amount of a compound that, when administered, is sufficient to prevent the occurrence of, or alleviate to some extent, one or more symptoms of the condition or disorder being treated. The therapeutically effective amount will vary depending on the compound, the disorder or condition and its severity, and the age, weight, etc., of the mammal being treated.

[0026] The term "pharmaceutically acceptable salts" in this disclosure includes salts of compounds of the present disclosure prepared using relatively non-toxic acids or bases, depending on the particular substituents found in the compounds described herein. When a compound of the present disclosure contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired base, either neat or in a suitable inert solvent. For example, salts can be derived from pharmaceutically acceptable inorganic bases, including aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, zinc, and the like. For example, salts may be derived from pharmaceutically acceptable organic bases, including salts of primary, secondary, and tertiary amines, including substituted amines, cyclic amines, naturally occurring amines, and the like, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When a compound of the present disclosure contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts are those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carboxylic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid, and the like, as well as relatively non-toxic acids such as acetic acid, propionic acid, isobutyric acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and the like. Also included are salts of amino acids, such as arginates, and salts of organic acids such as glucuronic acid or galactunoric acid (see, e.g., Berge, SM et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 66:1-19, 1977). Certain specific compounds of the present disclosure contain both basic and acidic functional groups, allowing the compounds to be converted into either base or acid addition salts.

[0027] In some embodiments, the neutral forms of the compounds are regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but the salts are otherwise equivalent to the parent form of the compound for purposes of this disclosure.

[0028] A "subject" is defined herein to include animals, such as mammals, including, but not limited to, primates (e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, etc. In some embodiments, the subject is a human.

[0029] The terms "treat," "treating," "treatment," and grammatical variations thereof, as used in this disclosure, include partially or completely delaying, alleviating, mitigating, or reducing the intensity, progression, or worsening of one or more associated symptoms of a disorder or condition, and / or alleviating, alleviating, or preventing one or more causes of a disorder or condition. Treatment in accordance with the present disclosure may be applied prophylactically, preventatively, palliatively, or curatively.

[0030] The compounds of the present disclosure may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may contain unnatural proportions of isotopes, such as deuterium ( 2 H), tritium (3 H), iodine-125( 125 I) or carbon-14 ( 14 C). All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are intended to be encompassed within the scope of the present disclosure.

[0031] The present inventors have developed compounds of formula I and II and have developed efficient methods for preparing compounds of formula I and II. [ka] [ka]

[0032] To determine whether the compounds are suitable A2aR antagonists, the compounds of Formulas I, II, III, and IV were subjected to GPCR screening using A2aR as the target. Screening was performed using the GPCR screening and profiling service provided by Eurofins Discover X Corporation. For comparison, NECA and SCH 442416 were also screened. The results are shown in Figures 1 to 6 and in the experiment.

[0033] Screening results showed that these compounds possess substantial A2aR antagonist properties.

[0034] Robert D. Leone, Ying-Chun Lo, and Jonathan D. Powell, "A2aR antagonists: Next generation checkpoint blockade for cancer immunotherapy," Comput Struct Biotechnol J. 2015;13:265-272, incorporated herein by reference in its entirety, discloses certain A2aR antagonists that may be useful in cancer immunotherapy. Thus, the compounds described herein are useful for treating cancer. Domenici MR et al., "Adenosine A2A receptor as a potential therapeutic target in neuropsychiatric disorders," Pharmacol Res 2019;147:104338, incorporated herein by reference in its entirety, discloses certain A2aR antagonists that may be useful for treating Alzheimer's disease, Parkinson's disease, attention-deficit hyperactivity disorder, fragile X syndrome, depression, and anxiety. Thus, the compounds described herein are useful for treating neurological disorders. Cronstein B. "Adenosine receptors and fibrosis: a translational view", F1000 Biol Rep. 2011;3:21, incorporated herein by reference in its entirety, discloses certain A2aR antagonists that may be useful in treating fibrosis, particularly liver fibrosis (e.g., NASH) and skin fibrosis (e.g., scleroderma).

[0035] One or more of the compounds described herein are useful for treating cancer, depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's or Parkinson's.

[0036] In some embodiments, the compounds of the present disclosure are useful in their pure form. In some embodiments, the compounds of the present disclosure are useful as pharmaceutical compositions prepared with a therapeutically effective amount of a compound as defined herein and a pharmaceutically acceptable excipient, such as a carrier or diluent.

[0037] In some embodiments, the compound is administered systemically, for example, orally in combination with a pharmaceutically acceptable vehicle, such as an inert excipient or an assimilable edible carrier, or by inhalation or insufflation. They may be enclosed in hard or soft-shelled gelatin capsules, compressed into tablets, or directly incorporated into the food of the patient's diet. For oral therapeutic administration, the compound may be combined with one or more additives and used in the form of ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, wafers, etc. The compound may be combined with a fine inert powder carrier and inhaled or insufflated by the subject. The percentage of compositions and preparations may, of course, vary and may be a suitable percentage of the mass of a given unit dosage form. The amount of the compound in such therapeutically useful compositions is such that an effective dosage level can be obtained.

[0038] In some embodiments, tablets, troches, pills, capsules, etc. may also contain: binders such as tragacanth gum, acacia, cornstarch, or gelatin; additives such as dicalcium phosphate; disintegrating agents such as cornstarch, potato starch, alginic acid, etc.; lubricants such as magnesium stearate; and sweeteners such as sucrose, fructose, lactose, or aspartame, or flavoring agents such as peppermint, wintergreen oil, or cherry flavoring. It is understood that capsules may contain, in addition to materials of the above type, a liquid carrier such as vegetable oil or polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For example, tablets, pills, or capsules may be coated with gelatin, wax, shellac, sugar, etc. Syrup or elixir may contain active compound, sucrose or fructose as sweetener, methyl and propylparaben as preservative, dye, and flavoring agent, such as cherry or orange flavor.It is understood that any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amount used.In some embodiments, compound is incorporated into sustained-release preparations and devices.For example, compound can be incorporated into sustained-release capsules, sustained-release tablets and sustained-release pills.

[0039] In some embodiments, the compound is administered intravenously or intraperitoneally by infusion or injection. A solution of the compound may be prepared in water, optionally mixed with a non-toxic surfactant. In some embodiments, dispersions are prepared in glycerol, liquid polyethylene glycol, triacetin, or mixtures thereof, or in oils. In some embodiments, these preparations contain a preservative to prevent the growth of microorganisms under normal storage and use conditions.

[0040] In some embodiments, pharmaceutical dosage forms for injection or infusion are sterile aqueous solutions or dispersions or sterile powders containing compounds, optionally encapsulated in liposomes, suitable for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions. In all cases, the ultimate dosage form should be sterile, fluid, and stable under the conditions of manufacture and storage. In some embodiments, the liquid carrier or vehicle is a solvent or liquid dispersion medium, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.), vegetable oils, non-toxic glyceryl esters, and suitable mixtures thereof. In some embodiments, proper fluidity is maintained by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions, or by the use of surfactants. In some embodiments, various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc., are used to prevent the action of microorganisms. In some embodiments, isotonic agents, such as sugars, buffers, or sodium chloride, are included. In some embodiments, agents delaying absorption, for example, aluminum monostearate or gelatin, are used to prolong absorption of the injectable compositions.

[0041] In some embodiments, sterile injectable solutions are prepared by incorporating the compound in the required amount in an appropriate solvent, optionally with some of the other ingredients enumerated above, as needed, followed by filtered sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the compositions may be vacuum-dried and / or freeze-dried to yield a powder of the active ingredient plus any additional desired ingredients present in the previously sterile-filtered solution.

[0042] In some embodiments, for topical administration, the compound is applied in pure form, hi some embodiments, the compound is administered to the skin as a composition or formulation in combination with a dermatologically acceptable carrier, which may be a solid or liquid.

[0043] In some embodiments, the solid carrier comprises a finely divided solid, such as talc, clay, microcrystalline cellulose, silica, alumina, etc. In some embodiments, the solid carrier comprises non-toxic polymeric nanoparticles or microparticles. In some embodiments, the liquid carrier comprises water, alcohol, or glycol, or a water / alcohol / glycol blend, in which the compound can be dissolved or dispersed at effective levels, optionally with the aid of a non-toxic surfactant. In some embodiments, adjuvants, such as fragrances and additional antimicrobial agents, are added to optimize the properties for a given use. The resulting liquid composition can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using a pump-action or aerosol sprayer.

[0044] In some embodiments, thickening agents such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified cellulose, or modified mineral materials are used with a liquid carrier to form spreadable pastes, gels, ointments, soaps, etc. for direct application to the user's skin.

[0045] In some embodiments, the compound is formulated in a lyophilized form for parenteral administration. In some embodiments, the lyophilized formulation is reconstituted by adding water or other aqueous medium, and then further diluted with a suitable excipient before use. In some embodiments, the liquid formulation is a buffered isotonic aqueous solution. In some embodiments, the excipient is isotonic saline, 5% dextrose in water, and buffered sodium or ammonium acetate solution. Pharmaceutically acceptable solid or liquid additives may be added to strengthen or stabilize the composition or to facilitate the preparation of the composition.

[0046] In some embodiments, the pharmaceutical compositions may additionally contain, in addition to the compounds described herein, one or more other pharmacologically active agents.

[0047] In some embodiments, useful dosages of the compounds can be determined by comparing their in vitro and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; see, for example, U.S. Patent No. 4,938,949, incorporated herein by reference.

[0048] It will be appreciated that the amount of compound required for use in treatment will vary not only with the particular salt selected, but also with the route of administration, the nature of the condition being treated and the age and condition of the patient, and will ultimately be at the discretion of the attending physician or clinician.

[0049] In some embodiments, the effective dosage and route of administration of the disclosed agents are conventional. The exact amount of the compound (effective dose) may vary from subject to subject, depending, for example, on the species, age, weight, and general or clinical condition of the subject, the severity or mechanism of any disorder being treated, the particular compound or vehicle used, the method and schedule of administration, etc. In some embodiments, the therapeutically effective dose is determined empirically by conventional procedures known to those of skill in the art. For example, those skilled in the art may refer to *The Pharmacological Basis of Therapeutics*, edited by Goodman and Gilman, Macmillan Publishing Co., New York. In some embodiments, the effective dose is initially estimated either in cell culture assays or in a suitable animal model. In some embodiments, animal models are used to determine appropriate concentration ranges and routes of administration. In some embodiments, such information is then used to determine useful doses and routes for administration in humans. In some embodiments, the therapeutic dose is selected by analogy to the dosage for a comparable therapeutic agent.

[0050] For example, dosages may be in the range of about 0.001 to about 100 mg per kg of body weight per day, e.g., about 0.01 to about 100 mg per kg of body weight per day, e.g., greater than about 0.1 mg per kilogram of recipient body weight per day, or in the range of about 1 to about 10 mg per kilogram of recipient body weight per day. For example, suitable doses may be about 0.3 mg, 0.7 mg, 1 mg, 10 mg, or 50 mg per kg of body weight per day.

[0051] In some embodiments, the compound is administered in unit dosage form, for example, containing 0.05 to 10000 mg, 0.5 to 10000 mg, 5 to 1000 mg, or about 100 mg of active ingredient per unit dosage form.

[0052] In some embodiments, the compound is administered to achieve a peak plasma concentration of, for example, about 0.5 to about 75 μM, about 1 to 50 μM, about 2 to about 30 μM, or about 5 to about 25 μM. Exemplary desirable plasma concentrations include at least 0.25, 0.5, 1, 5, 10, 25, 50, 75, 100, or 200 μM or less. For example, plasma levels may be about 1 to 100 micromolar or about 10 to about 25 micromolar. In some embodiments, this is achieved by intravenous injection of a 0.05 to 5% solution of the compound, optionally in saline, or orally administered as a bolus containing about 1-100 mg of the compound. In some embodiments, desirable blood levels are maintained by continuous infusion to provide about 0.00005-5 mg / kg body weight per hour, e.g., at least 0.00005, 0.0005, 0.005, 0.05, 0.5, or 5 mg / kg / hour or less. In some embodiments, such levels are obtained by intermittent infusion containing about 0.0002-20 mg / kg body weight, e.g., at least 0.0002, 0.002, 0.02, 0.2, 2, 20, or 50 mg / kg body weight or less of the compound.

[0053] In one embodiment, the amount of Purine Compounds used is between 0.1 and 5 mg per kg of subject per day. In a preferred embodiment, the amount of Purine Compounds used is between 0.2 and 1.3 mg per kg of subject per day. In a further preferred embodiment, the amount of Purine Compounds used is about 0.3 to 0.7 mg per kg of subject per day.

[0054] In some embodiments, the compounds are presented in a single dose or as divided doses administered at appropriate intervals, for example, two, three, four or more sub-doses per day, which in some embodiments are themselves further divided, for example, into several discrete, spaced administrations, such as multiple inhalations from an insufflator.

[0055] In some embodiments, pharmaceutical compositions of the present disclosure are packaged in a container with a label or instructions, or both, directing use of the pharmaceutical composition in treating an indicated disease.

[0056] It is understood that one or more compounds of the present disclosure may be used independently or in any combination thereof. [Example]

[0057] Example 1 The compounds of Formulas I, II, III, and IV were screened for GPCR activity using A2aR as the target. For comparison, NECA and SCH 442416 were also screened. NECA is an A2aR agonist. SCH 442416 is an A2aR antagonist.

[0058] Assay design: calcium mobilization Cell handling Cell lines were expanded from freezer stocks according to standard procedures. Cells were seeded in a total volume of 20 μL into black-walled, clear-bottom, poly-D-lysine-coated 384-well microplates and incubated at 37°C for the appropriate time before testing.

[0059] Dye loading Assays were performed in 1x dye loading buffer consisting of 1x dye, 1x additive A, and 2.5mM probenecid in HBSS / 20mM Hepes. Probenecid was freshly prepared. Cells were loaded with dye prior to testing. Media was aspirated from cells and replaced with 20µL of dye loading buffer. Cells were incubated at 37°C for 30-60 minutes.

[0060] Agonist Format For agonist determination, cells were incubated with the sample to induce a response. After dye loading, cells were removed from the incubator and 10 μL of HBSS / 20 mM Hepes was added. When performing an agonist dose curve to define the EC80 for subsequent antagonist assays, 3× vehicle was included in the buffer. Cells were incubated in the dark at room temperature for 30 minutes to equilibrate the plate temperature. Intermediate dilutions of the sample stock were performed to generate 4× samples in assay buffer. Compound agonist activity was measured using a FLIPR Tetra (MDS). Calcium mobilization was monitored over 2 minutes, and 10 μL of the 4× sample in HBSS / 20 mM Hepes was added to the cells 5 seconds into the assay.

[0061] Allosteric Modulation Format For allosteric determination, cells were preincubated with sample followed by agonist challenge at EC20 concentration. Intermediate dilutions of sample stock were performed to generate 3x samples in assay buffer. After dye loading, cells were removed from the incubator and 10 μL of 3x sample was added. Cells were incubated in the dark at room temperature for 30 minutes to allow plate temperature equilibration. Vehicle concentration was 1%. Compound allosteric activity was measured with a FLIPR Tetra (MDS). Calcium mobilization was monitored over 2 minutes, and 10 μL of 4x EC20 agonist in HBSS / 20 mM Hepes was added to the cells 5 seconds into the assay.

[0062] Antagonist Format For antagonist determination, cells were preincubated with sample followed by agonist challenge at an EC80 concentration. Intermediate dilutions of sample stock were performed to generate 3x samples in assay buffer. After dye loading, cells were removed from the incubator and 10 μL of 3x sample was added. Cells were incubated in the dark at room temperature for 30 minutes to allow plate temperature equilibration. Vehicle concentration was 1%. Compound antagonist activity was measured with a FLIPR Tetra (MDS). Calcium mobilization was monitored over 2 minutes, and 10 μL of EC80 agonist in HBSS / 20 mM Hepes was added to the cells 5 seconds into the assay.

[0063] Data analysis Compound activity was analyzed using the CBIS data analysis suite (ChemInnovation, CA). For agonist mode assays, percent activity is calculated using the following formula: % Activity = 100% x (mean RFU of test sample - mean RFU of vehicle control) / (mean MAX RFU control ligand - mean RFU of vehicle control).

[0064] For positive allosteric mode assays, the percent modulation is calculated using the following formula: % Modulation = 100% x ((mean RFU of test sample - mean RFU of EC20 control) / (mean RFU of MAX control ligand - mean RFU of EC20 control)).

[0065] For antagonist and negative allosteric modulation mode assays, the percentage of inhibition is calculated using the following formula: % Inhibition = 100% x (1 - (mean RFU of test sample - mean RFU of vehicle control) / (mean RFU of EC80 control - mean RFU of vehicle control)).

[0066] Figures 1 to 6 present the results of a G protein-coupled receptor (GPCR) screen using A2aR as a target by Eurofins Discover X Corporation's GPCR screening and profiling service.

[0067] In the screen, activation of A2aR led to calcium mobilization, which was monitored using a calcium-sensitive dye: upon calcium release, the fluorescence of the dye increased, and the increase was measured in real time.

[0068] Figures 1 to 6 show percent responded (Y) versus concentration in μM (X). Compounds were tested in antagonist mode using the required GPCR biosensor assay. For antagonist assays, data were normalized to the maximum and minimum responses observed in the presence of the EC80 ligand and vehicle. The following EC80 concentrations were used: ADORA2A calcium flux: 0.021 μM NECA.

[0069] Figure 1 shows the results for the compound of formula I. The half-maximal inhibitory concentration (IC 50 ) was 0.08851 μM.

[0070] Figure 2 shows the results for compounds of formula I. IC 50 was 0.07545 μM.

[0071] Figure 3 shows the results for the compound of formula III. IC 50 was 0.1438 μM.

[0072] Figure 4 shows the results for the compound of formula IV, istradefylline. IC 50 was 0.0667 μM.

[0073] Figure 5 shows the results for NECA. 50 ) was 0.007637 μM.

[0074] Figure 6 shows the results for SCH 442416. IC 50 was 0.0113 μM.

[0075] Summary of results [Table 1]

[0076] Example 2 Pharmacokinetic data for compounds of formula I, II, III and IV.

[0077] Male CD-I mice were given either a bolus of 1 mg / kg intravenously or 3 mg / kg orally of the indicated compounds and plasma or brain concentrations were measured.

[0078] Plasma concentrations of compounds of formula I following a 1 mg / kg bolus intravenous dose. [Table 2]

[0079] Brain concentrations of compounds of formula I following a 1 mg / kg bolus intravenous dose. [Table 3]

[0080] Brain / plasma concentration ratio after 1 mg / kg bolus intravenous administration of 1a. [Table 4]

[0081] FIG. 7 is a graph showing plasma and brain concentrations of the compound of formula I over time.

[0082] Plasma concentrations of compounds of formula I following oral administration of 3 mg / kg. [Table 5]

[0083] FIG. 8 is a graph showing the plasma concentration of a compound of formula I over time following administration of 1 mg / kg intravenously or 3 mg / kg orally.

[0084] Summary of plasma PK parameters for the compound of formula I. [Table 6]

[0085] Summary of brain PK parameters for the compound of formula I. [Table 7]

[0086] For the compound of Formula I, the measured dosing solution concentrations were 0.203 mg / ml and 0.296 for the intravenous and oral formulations, respectively.

[0087] where: C0 Concentration extrapolated to time zero after intravenous dose t max Time at which maximum concentration is observed C max Maximum observed concentration Apparent t 1 / 2 Apparent terminal half-life AUC 0~tlast Area under the concentration versus time curve from time 0 to the time of the last measurable concentration AUC 0~inf Area under the concentration versus time curve from time 0 to infinity CL Full Body Clearance MRT 0~inf Average residence time from time zero to infinity V ss Steady-state volume of distribution F oral bioavailability = (dose iv ×AUC po ) / (dose po ×AUC iv ) x 100

[0088] Plasma concentrations of the compound of formula II following a 1 mg / kg bolus intravenous dose. [Table 8]

[0089] Brain concentrations of the compound of formula II following a 1 mg / kg bolus intravenous dose. [Table 9]

[0090] Brain / plasma concentration ratio following 1 mg / kg bolus intravenous administration of the compound of formula II. [Table 10]

[0091] FIG. 9 is a graph showing the plasma and brain concentrations of the compound of formula II over time.

[0092] Plasma concentrations of the compound of formula II following oral administration of 3 mg / kg. [Table 11]

[0093] FIG. 10 is a graph showing the plasma concentration of a compound of formula II over time following administration of 1 mg / kg intravenously or 3 mg / kg orally.

[0094] Summary of plasma PK parameters for the compound of formula II. [Table 12]

[0095] Summary of brain PK parameters for the compound of formula II. [Table 13]

[0096] For the compound of Formula II, the measured dosing solution concentrations were 0.193 mg / ml and 0.299 for the intravenous and oral formulations, respectively.

[0097] Plasma concentrations of Formula III (SD-007) following a 1 mg / kg bolus intravenous dose. [Table 14]

[0098] Brain concentrations of the compound of formula III following a 1 mg / kg bolus intravenous dose. [Table 15]

[0099] Brain / plasma concentration ratio following 1 mg / kg bolus intravenous administration of the compound of formula III. [Table 16]

[0100] FIG. 11 is a graph showing the plasma and brain concentrations of the compound of formula III over time.

[0101] Plasma concentrations of the compound of formula III following oral administration of 3 mg / kg. [Table 17]

[0102] FIG. 12 is a graph showing the plasma concentration of the compound of formula III over time following administration of 1 mg / kg intravenously or 3 mg / kg orally.

[0103] Summary of plasma PK parameters for the compound of formula III. [Table 18]

[0104] Summary of brain PK parameters for the compound of formula III. [Table 19]

[0105] For the compound of formula III, the measured dosing solution concentrations were 0.196 mg / ml and 0.282 for the intravenous and oral formulations, respectively.

[0106] Plasma concentrations of istradefylline following a 1 mg / kg bolus intravenous administration. [Table 20]

[0107] Brain concentrations after a 1 mg / kg bolus intravenous administration of istradefylline. [Table 21]

[0108] Brain / plasma concentration ratio after a 1 mg / kg bolus intravenous administration of istradefylline. [Table 22]

[0109] FIG. 13 is a graph showing the plasma and brain concentrations of the compound of formula IV over time.

[0110] Plasma concentrations of istradefylline following oral administration of 3 mg / kg. [Table 23]

[0111] Values ​​in italics are below the lower limit of quantitation (BLQ, 0.5 ng / mL) but are included in the calculation; BLQ indicates below the lower limit of quantitation (0.5 ng / mL); n / a indicates not applicable.

[0112] FIG. 14 is a graph showing the plasma concentration over time of the compound of Formula IV, istradefylline, following administration of 1 mg / kg intravenously or 3 mg / kg orally.

[0113] Summary of plasma PK parameters for istradefylline. [Table 24]

[0114] Summary of brain PK parameters for istradefylline. [Table 25]

[0115] For the compound of formula IV, the measured dosing solution concentrations were 0.221 mg / ml and 0.325 for the intravenous and oral formulations, respectively.

[0116] Example 3 Compounds of Formula I, II, III and IV were tested in depression studies using mice and the forced swim test, and the results are shown in Figures 15 and 16.

[0117] The forced swim test, also known as the behavioral despair test, is used to test for depression-like behavior. The test involves placing rats or mice in a cylinder filled with water. "Floating behavior" (when the animal remains largely immobile and keeps its head above water) is used as a parameter to analyze "hopelessness" and therefore depression-like behavior. Rodents given antidepressants swim longer than controls. Immobility time is reduced by various antidepressants.

[0118] Forty male CD-I mice (N = 8 per group) were orally treated with either vehicle (0.3% Tween 80) or a test article (1 mg / kg) of Formula I (Target 1a), II (Target 1b), III (SD-007), or IV (Istradefylline, KW-6002) one hour prior to testing. At t = 0, mice were placed in water-filled glass cylinders. After a period of vigorous activity, mice would adopt a characteristic immobile posture that was easily scored. The latency to first immobility was recorded (in seconds) over the 6-minute test session. The duration of immobility (in seconds) during the final 4 minutes of testing was also measured.

[0119] Formula II statistically significantly delayed the time to immobility (one-way ANOVA with Dunnett's multiple comparison test). Similar effects of Formulas II and IV were seen on immobility time (2-6 minutes). Thus, the efficacy of these compounds in reducing depression-like behavior was demonstrated.

[0120] The embodiments of the present disclosure described above are intended to be examples only. The present disclosure may be embodied in other specific forms. Alterations, modifications, and variations to the present disclosure may be made without departing from the intended scope of the present disclosure. While the systems, devices, and processes disclosed and illustrated herein may include a specific number of elements / components, the systems, devices, and assemblies may be modified to include additional or fewer such elements / components. For example, while any of the disclosed elements / components may be referred to in the singular, the embodiments disclosed herein may be modified to include multiple such elements / components. Features selected from one or more of the above-described embodiments may be combined to create alternative embodiments not expressly described. All values ​​and subranges within the disclosed ranges are also disclosed. The subject matter described herein is intended to cover and encompass all suitable modifications in technology. All references cited are incorporated herein by reference in their entirety.

Claims

1. formula 【Chemistry 1】 or a pharmaceutically acceptable salt thereof.

2. formula 【Chemistry 2】 or a pharmaceutically acceptable salt thereof.

3. A pharmaceutical composition for treating depression, said composition comprising a compound of formula 【Transformation 3】 and pharmaceutically acceptable salts thereof.

4. A pharmaceutical composition for treating cancer, said composition comprising a compound of the formula 【Chemistry 4】 and pharmaceutically acceptable salts thereof.

5. 1. A pharmaceutical composition for treating one or more of depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's and Parkinson's, said composition comprising a compound of the formula 【Transformation 5】 and pharmaceutically acceptable salts thereof.

6. The pharmaceutical composition according to claims 3 to 5, wherein the administration of the composition is oral.

7. The pharmaceutical composition according to claims 3 to 5, wherein the administration of the composition is by injection.

8. 8. The pharmaceutical composition of claims 3 to 7, wherein the administration of the composition is accompanied by a pharmaceutically acceptable excipient.

9. A pharmaceutical composition for treating depression, said composition comprising a compound of formula 【Transformation 6】 and pharmaceutically acceptable salts thereof.

10. formula 【Transformation 7】 or a pharmaceutically acceptable salt thereof, for treating depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's or Parkinson's.

11. 1. A pharmaceutical composition for treating depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's or Parkinson's, said composition comprising a compound of formula 【Transformation 8】 and pharmaceutically acceptable salts thereof.

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