Purine compounds for treating disorders

Abstract

One objective is to provide an A2aR antagonist suitable for treating neurological disorders, fibrosis-related diseases (NASH and scleroderma), and cancer. [Solution] Substituted purine compounds of formulas (I) to (III) and salts thereof act as adenosine A2a receptor (A2aR) antagonists for cancer immunotherapy, depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's disease 5, and Parkinson's disease. Pharmaceutical compositions containing such compounds and methods of their use in treating depression are also taught. JPEG2026074001000043.jpg70170

Description

[Technical Field]

[0001] Field of the present invention The present invention relates, in general, to substituted purine compounds and salts thereof that act as adenosine A2a receptor (A2aR) antagonists for cancer, depression, anxiety, multiple sclerosis, NASH, scleroderma (scleroderma), ADHD, Alzheimer's disease, and Parkinson's disease, pharmaceutical compositions comprising such compounds, and methods of using them in the treatment of cancer, depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's disease, and Parkinson's disease. [Background technology]

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

[0003] A2aRs have been shown to play a regulatory role in the immune system. One A2aR antagonist, istradefylline, has been shown to reduce motor dysfunction and, in turn, improve 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 central nervous system disorders, neoplastic 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. [Overview of the project] [Problems that the invention aims to solve]

[0007] There is a need for an A2aR antagonist suitable for treating neurological disorders, fibrosis-related diseases (NASH and scleroderma), and cancer.

Means for Solving the Problem

[0008] Summary of the Invention The present inventors have found a certain purine compound useful as an A2aR antagonist for treating diseases such as depression.

[0009] In one aspect, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided.

Chemical Formula

[0010] In one aspect, a compound of formula (II) or a pharmaceutically acceptable salt thereof is provided.

Chemical Formula

[0011] In one aspect, a compound of formula (III) or a pharmaceutically acceptable salt thereof is provided.

Chemical Formula

[0012] In one aspect, 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 can be useful for treating cancer, depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's and Parkinson's. [ka]

[0013] In one embodiment, SCH 442416, Formula (V) 2-(2-furanyl)-7-[3-(4-methoxyphenyl)propyl]-7H-pyrazolo[4,3-e][1,2,4]triazolo[1,5-c]pyrimidine-5-amine, 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 disease, and Parkinson's disease. [ka]

[0014] The disclosure also includes pharmaceutical compositions comprising one or more of the compounds of formulas I, II, and III and pharmaceutically acceptable salts thereof in therapeutically effective amounts, as well as pharmaceutically acceptable excipients.

[0015] This disclosure further includes methods for treating depression using one or more compounds of formulas 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.

[0016] This disclosure further includes methods for treating cancer, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's disease, or Parkinson's disease using one or more compounds of formulas 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 disease, and Parkinson's disease.

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

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

[0019] This disclosure includes a method comprising the steps of preparing at least one such compound, measuring the inhibition of A2aR activity for the compound, and determining whether the inhibition is above an expected level. Brief explanation of the drawing [Brief explanation of the drawing]

[0020] [Figure 1] This graph shows the GPCR screening results for the compound of formula I. [Figure 2] This graph shows the GPCR screening results for the compound of formula II. [Figure 3] This graph shows the GPCR screening results for compound SD-007, represented by formula III. [Figure 4] This graph shows the GPCR screening results for the compound istradefyline, represented by formula IV. [Figure 5] This graph shows the GPCR screening results for 5'-N-ethylcarboxamide adenosine (NECA). [Figure 6] This graph shows the GPCR screening results for SCH 442416. [Figure 7] This graph shows the plasma and brain concentrations of the compound of formula I over time. [Figure 8] This graph shows the plasma concentration over time after administration of the compound of formula I at a dose of 1 mg / kg intravenously or 3 mg / kg orally. [Figure 9] This graph shows the plasma and brain concentrations of the compound of formula II over time. [Figure 10] This graph shows the plasma concentration over time after administration of the compound of formula II at a dose of 1 mg / kg intravenously or 3 mg / kg orally. [Figure 11] This graph shows the plasma and brain concentrations of the compound SD-007, represented by formula III, over time. [Figure 12] This graph shows the plasma concentration over time of the compound SD-007, of formula III, after intravenous administration of 1 mg / kg or oral administration of 3 mg / kg. [Figure 13] This graph shows the plasma and brain concentrations of the compound istradefylline (formula IV) over time. [Figure 14] This graph shows the plasma concentration over time after administration of the compound of formula IV at a dose of 1 mg / kg intravenously or 3 mg / kg orally. [Figure 15] This graph compares the time from incubation to immobility for various compounds. [Figure 16] This graph compares the immobilization times of various compounds. [Modes for carrying out the invention]

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

[0022] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which this disclosure pertains.

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

[0024] When used herein, the term "including" should be understood to mean that the following list is not exhaustive and may, as appropriate, include or exclude any other additional preferred items, such as one or more further features, ingredients and / or raw materials.

[0025] The terms "pharmaceutical effective dose," "therapeutic effective dose," or "therapeutic dose," and "effective dose" refer to the amount of a compound of interest that would elicit a biological or medical response in a tissue, system, animal, or human, as sought by researchers, veterinarians, physicians, or other clinicians. The term "therapeutic effective dose" includes the amount of a compound that, when administered, is sufficient to prevent or, to some extent, alleviate the occurrence of one or more symptoms of the condition or disorder being treated. The therapeutic effective dose will vary depending on the compound, the disorder or condition and its severity, as well as the age, weight, etc., of the mammal being treated.

[0026] In this disclosure, the term “pharmaceutically acceptable salt” includes salts of the compounds of this disclosure prepared with relatively non-toxic acids or bases, depending on the specific substituents found in the compounds described herein. If the compounds of this disclosure contain relatively acidic functional groups, the base addition salt may be obtained by contacting the neutral form of such compound with a sufficient amount of the desired base, either undiluted or in a suitable inert solvent. For example, the salt may be derived from pharmaceutically acceptable inorganic bases, including aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganese, manganese, potassium, sodium, zinc, etc. For example, the salts may be derived from pharmaceutically acceptable organic bases, including salts of primary, secondary, and tertiary amines, such as substituted amines, cyclic amines, and naturally occurring amines, including arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydravamin, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. If the compounds of this disclosure contain relatively basic functional groups, the acid addition salts may be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either undiluted or in a suitable inert solvent.Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carboxylic acid, monohydrogencarbonic, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid, or phosphorous acid, as well as salts derived from relatively non-toxic organic 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, and methanesulfonic acid. This also includes salts of amino acids, such as alginates, and salts of organic acids, such as glucuronic acid or galactunoric acid (see, for example, Berge, SM et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 66:1-19, 1977). Certain specific compounds in this disclosure contain both basic and acidic functional groups, allowing the compounds to be converted to either a base or an acid addition salt.

[0027] In some embodiments, the neutral form of the compound is regenerated by contacting the salt with a base or acid and isolating the parent compound in a 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 salt is otherwise equivalent to the parent form of the compound for the purposes of this disclosure.

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

[0029] The terms “to treat,” “to treat,” “treatment,” and their grammatical variations, as used in this disclosure, include partially or completely delaying, mitigating, or reducing the intensity, progression, or worsening of one or more accompanying symptoms of a disorder or condition, and / or mitigating, reducing, or preventing the cause of one or more of the disorders or conditions. Treatments pursuant to this disclosure may be applied preventively, defensively, pallatively, or remedially.

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

[0031] The inventors have developed compounds of formulas I and II, and have developed an efficient method for preparing compounds of formulas I and II. [ka] [ka]

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

[0033] The 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, is incorporated herein by reference in its entirety and discloses certain A2aR antagonists that may be useful in cancer immunotherapy. Therefore, the compounds described herein are useful for treating cancer. Domenici MR et al., "Adenosine A2A receptor as potential therapeutic target in neuropsychiatric disorders," Pharmacol Res 2019; 147: 104338, is incorporated herein by reference in its entirety and discloses certain A2aR antagonists that may be useful for treating Alzheimer's disease, Parkinson's disease, attention-deficit / hyperactivity disorder, fragility X syndrome, depression, and anxiety. Therefore, 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, which is incorporated herein by reference in its entirety, discloses certain A2aR antagonists that may be useful in treating fibrosis, particularly hepatic fibrosis (e.g., NASH) and cutaneous 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 disease, or Parkinson's disease.

[0036] In some embodiments, the compounds of the Disclosure are useful in their pure form. In some embodiments, the compounds of the Disclosure are useful as pharmaceutical compositions prepared with therapeutically effective amounts of the compounds as defined herein and pharmaceutically acceptable additives, such as carriers or excipients.

[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 absorbable edible carrier, or by inhalation or inhalation. They may be encapsulated in hard or soft-shelled gelatin capsules, compressed into tablets, or directly incorporated into the patient's diet. For oral therapeutic administration, the compound may be combined with one or more excipients and may be used in the form of ingestible tablets, oral tablets, lozenges, capsules, elixirs, suspensions, syrups, cachets, etc. The compound may be combined with a fine inert powder carrier and administered by inhalation or inhalation depending on the subject. The percentages of compositions and preparations may, of course, vary and may be preferred percentages 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, lozenges, pills, capsules, etc., also contain: binders, e.g., tragacanth gum, acacia, corn starch, or gelatin; additives, e.g., dicalcium phosphate; disintegrants, e.g., corn starch, potato starch, alginic acid, etc.; lubricants, e.g., magnesium stearate; and sweeteners, e.g., sucrose, fructose, lactose, or aspartame, or flavorings, e.g., peppermint, wintergreen oil, or cherry flavorings. Capsules may contain, in addition to the above types of materials, a liquid carrier, e.g., 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, or sugar, etc. The syrup or elixir may contain the active compound, sucrose or fructose as a sweetener, methyl and propylparaben as preservatives, dyes, and flavorings, such as cherry or orange flavorings. It is understood that any materials used in preparing any unit dosage form should be pharmaceutically acceptable and substantially nontoxic in the amounts used. In some embodiments, the compound is incorporated into sustained-release preparations and devices. For example, the compound may 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 injection or infusion. Solutions of the compound may be prepared in water mixed with a non-toxic surfactant, if applicable. In some embodiments, dispersions are prepared in glycerol, liquid polyethylene glycol, triacetin or a mixture thereof, or in oil. In some embodiments, these preparations contain preservatives to prevent microbial growth under normal storage and use conditions.

[0040] In some embodiments, the pharmaceutical dosage form for injection or infusion is a sterile aqueous solution or dispersion or sterile powder, which may contain compounds adapted for immediate preparation of sterile injection or infusion solutions or dispersions, sometimes encapsulated in liposomes. In all cases, the final dosage form should be sterile, fluid, and stable under manufacturing and storage conditions. In some embodiments, the liquid carrier or vehicle is a solvent or liquid dispersion medium, and includes, 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, reasonable fluidity is maintained by liposome formation, by maintaining 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 microbial activity. In some embodiments, isotonic agents, such as sugars, buffers, or sodium chloride, are included. In some embodiments, an absorption-delaying agent, such as aluminum monostearate or gelatin, is used to extend the absorption of the injectable composition.

[0041] In some embodiments, the sterile injection solution is prepared by combining the required amount of compound in a suitable solvent, optionally together with some of the other raw materials listed above, and subsequently by sterilization by filtration, if necessary. In the case of a sterile powder for the preparation of the sterile injection solution, the composition may be vacuum-dried and / or freeze-dried to produce a powder of the active raw material plus any additional desired raw materials present in the pre-sterilized filtered solution.

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

[0043] In some embodiments, the solid carrier comprises a pulverized solid, such as talc, clay, microcrystalline cellulose, silica, or alumina. In some embodiments, the solid carrier comprises non-toxic polymeric nanoparticles or fine particles. In some embodiments, the liquid carrier comprises water, alcohol, glycol, or a water / alcohol / glycol blend, in which the compound may be dissolved or dispersed at an effective level, optionally utilizing a non-toxic surfactant. In some embodiments, adjuvants, such as fragrances and additional antimicrobial agents, are added to optimize properties for a given use. The resulting liquid composition may be applied from an absorbent pad, used to impregnate plasters and other bandages, or sprayed onto the affected area using a pump or aerosol sprayer.

[0044] In some embodiments, a thickener, such as a synthetic polymer, fatty acid, fatty acid salt and ester, fatty alcohol, modified cellulose, or modified mineral material, is used together with a liquid carrier to form a paste, gel, ointment, soap, etc., that can be spread and applied directly 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 the addition of water or another 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 a buffered sodium acetate or ammonium solution. Pharmaceutically acceptable solid or liquid additives may be added to enhance or stabilize the composition, or to facilitate the preparation of the composition.

[0046] In some embodiments, the pharmaceutical composition may contain one or more other pharmacological agents in addition to the compounds described herein.

[0047] In some embodiments, the effective dosage of a compound can be determined by comparing its in vitro and in vivo activities in animal models. Methods for extrapolating effective dosages in mice and other animals to humans are known in the Art; see, for example, U.S. Patent No. 4,938,949, which is incorporated herein by reference.

[0048] The amount of compound required for use in a procedure will vary not only with the specific salt selected, but also with the route of administration, the nature of the condition being treated, and the patient's age and condition, and will ultimately be at the discretion of the attending physician or clinician.

[0049] In some embodiments, the effective dose and route of administration of the active ingredients of this disclosure are conventional. The exact amount (effective dose) of the compound may vary depending on the subject, for example, the species, age, weight and systemic or clinical condition of the subject, the severity or mechanism of any disorder being treated, the specific compound or vehicle used, the method and scheduling of administration, etc. In some embodiments, the therapeutic effective dose is determined empirically by conventional procedures known to those skilled 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 first estimated in either a cell culture assay or a suitable animal model. In some embodiments, the animal model is used to determine an appropriate concentration range and route of administration. In some embodiments, such information is then used to determine a useful dose and route for administration in humans. In some embodiments, the therapeutic dose is selected by analogy to the dosage for an equivalent therapeutic agent.

[0050] For example, the dosage may be in the range of approximately 0.001 to approximately 100 mg per kg of body weight per day, for example, in the range of approximately 0.01 to approximately 100 mg, for example, more than approximately 0.1 mg per kilogram of body weight of the recipient per day, or in the range of approximately 1 to approximately 10 mg per kilogram of body weight of the recipient per day. For example, a preferred dose may be approximately 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 forms containing, for example, 0.05 to 10,000 mg, 0.5 to 10,000 mg, 5 to 1,000 mg, or about 100 mg of active raw material per unit dosage form.

[0052] In some embodiments, the compound is administered to achieve peak plasma concentrations, 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, the plasma level may be up to about 1 to 100 micromolar concentrations or up to about 10 to about 25 micromolar concentrations. In some embodiments, this is achieved, optionally by intravenous injection of a 0.05 to 5% solution of the compound in physiological saline, or by oral administration as a bolus containing about 1 to 100 mg of the compound. In some embodiments, the desired blood level is maintained by continuous infusion to provide about 0.00005 to 5 mg per kg of body weight per hour, for example, 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 to 20 mg of the compound per kg of body weight, for example, at least 0.0002, 0.002, 0.02, 0.2, 2, 20, or 50 mg or less of the compound per kg of body weight.

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

[0054] In some embodiments, the compound is presented as a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more subdose doses per day. In some embodiments, the subdose itself is further divided into, for example, several separate doses administered at intervals apart, such as multiple inhalations from an inhaler.

[0055] In some embodiments, the pharmaceutical compositions of the present disclosure are packaged in containers with labels or instructions, or both, that indicate the use of the pharmaceutical compositions in the treatment of a disease being indicated.

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

[0057] Example 1 Compounds of formulas I, II, III, and IV were screened for GPCR activity using A2aR as a 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 frozen stocks following standard procedures. Cells were seeded in a total volume of 20 μL into 384-well microplates coated with poly-D-lysine and black-walled, clear-bottomed plates, and incubated at 37°C for an appropriate pre-test time.

[0059] Dye loading The assay was performed in a 1x dye loading buffer consisting of 1x dye, 1x additive A, and 2.5mM probenecid in HBSS / 20mM Hepes. Probenicid was prepared fresh. The cells were loaded with the dye before the test. The culture medium was aspirated from the cells and replaced with 20 μL of dye loading buffer. The cells were incubated at 37°C for 30–60 minutes.

[0060] Agonist format To determine the agonist, cells were incubated with the sample to induce a response. After dye loading, the cells were removed from the incubator and 10 μL of HBSS / 20 mM Hepes was added. A 3× vehicle was included in the buffer when running an agonist dose curve to define EC80 for the subsequent antagonist assay. The 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 produce 4× samples in the assay buffer. Compound agonist activity was measured using FLIPR Tetra (MDS). Calcium recruitment was monitored for 2 minutes, and 4× samples in 10 μL of HBSS / 20 mM Hepes were added to the cells 5 seconds after the start of the assay.

[0061] Allosteric Modulation Format For allosteric determination, cells were pre-incubated with the sample, followed by agonist induction at EC20 concentration. Intermediate dilution of the sample stock was performed to produce 3× sample in assay buffer. After dye loading, cells were removed from the incubator and 10 μL of 3× sample was added. Cells were incubated in the dark at room temperature for 30 minutes to equilibrate plate temperature. Vehicle concentration was 1%. Allosteric activity of the compound was measured by FLIPR tetra (MDS). Calcium recruitment was monitored for 2 minutes, and 4× EC20 agonist in 10 μL of HBSS / 20 mM Hepes was added to the cells 5 seconds after the start of the assay.

[0062] Antagonist format To determine the antagonist, cells were pre-incubated with the sample, followed by agonist loading at EC80 concentration. Intermediate dilution of the sample stock was performed to produce 3× sample in assay buffer. After dye loading, cells were removed from the incubator and 10 μL of 3× sample was added. Cells were incubated in the dark at room temperature for 30 minutes to equilibrate plate temperature. Vehicle concentration was 1%. Compound antagonist activity was measured by FLIPR tetra (MDS). Calcium recruitment was monitored for 2 minutes, and EC80 agonist in 10 μL of HBSS / 20 mM Hepes was added to cells 5 seconds after the start of the assay.

[0063] Data Analysis Compound activity was analyzed using the CBIS Data Analysis Suite (ChemInnovation, CA). For the agonist mode assay, the activity percentage was calculated using the following formula: Activity % = 100% × (Average RFU of test sample - Average RFU of vehicle control) / (Average MAX RFU of control ligand - Average RFU of vehicle control).

[0064] For a positive allosteric mode assay, the modulation percentage is calculated using the following formula: Modulation % = 100% × ((Average RFU of test sample - EC20 average RFU of control) / (Average RFU of MAX control ligand - EC20 average RFU of control)).

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

[0066] Figures 1 to 6 present the results of G protein-coupled receptor (GPCR) screening targeting A2aR by the GPCR screening and profiling service of Eurofins Discover X Corporation.

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

[0068] Figures 1 to 6 show the percent reacted (Y) against concentration in μM (X). Compounds were tested in antagonist mode using the required GPCR biosensor assay. For the antagonist assay, data were normalized against 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 the compound 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. The half-maximal effective concentration (EC 50 ) was 0.007637 μM.

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

[0075] Summary of Results [Table 1]

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

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

[0078] Plasma concentration after intravenous administration of a 1 mg / kg bolus of the compound of formula I. [Table 2]

[0079] Brain concentration after intravenous administration of a 1 mg / kg bolus of the compound of formula I. [Table 3]

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

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

[0082] Plasma concentration after oral administration of 3 mg / kg of the compound of formula I. [Table 5]

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

[0084] Summary of plasma PK parameters for compounds 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 drug solution concentrations were 0.203 mg / ml and 0.296 mg / ml for intravenous and oral formulations, respectively.

[0087] Here, Concentration extrapolated to time zero after C0 intravenous dose t max Time at which maximum concentration is observed C max Maximum observed concentration Appearance t 1 / 2 Apparent terminal half-life AUC 0~tlast Area under the concentration-time curve from time 0 to the time of the last measurable concentration AUC 0~inf Concentration vs. Area under the Time Curve from Time 0 to Infinity CL Full Body Clearance MRT 0~inf Average dwell time from zero to infinity V ss Steady-state distributed volume F Oral bioavailability = (dose) iv ×AUC po ) / (dose po ×AUC iv ) × 100

[0088] Plasma concentration after intravenous administration of a 1 mg / kg bolus of the compound of formula II. [Table 8]

[0089] Brain concentration after intravenous administration of a 1 mg / kg bolus of the compound of formula II. [Table 9]

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

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

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

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

[0094] Summary of plasma PK parameters for compounds 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 drug solution concentrations were 0.193 mg / ml and 0.299 mg / ml for intravenous and oral formulations, respectively.

[0097] Plasma concentration after intravenous administration of a 1 mg / kg bolus of Formula III (SD-007). [Table 14]

[0098] Brain concentration after intravenous administration of a 1 mg / kg bolus of the compound of formula III. [Table 15]

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

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

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

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

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

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

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

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

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

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

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

[0110] Plasma concentration after oral administration of istradefylline at a dose of 3 mg / kg. [Table 23]

[0111] Italicized values ​​are included in the calculation even though they are below the limit of quantification (BLQ, 0.5 ng / mL). BLQ indicates a value below the limit of quantification (0.5 ng / mL), and n / a indicates that the calculation is not applicable.

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

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

[0114] An overview of brain PK parameters related to istradefylline. [Table 25]

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

[0116] Example 3 Compounds of formulas I, II, III, and IV were tested in mice and in depression studies using the forced swimming test. 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 depressive-like behaviors. The test involves placing a rat or mouse in a cylinder filled with water. "Floating behavior" (when the animal remains nearly motionless with its head above the water) is used as a parameter to analyze "despair" and therefore depressive-like behaviors. Rodents given antidepressants swim longer than controls. Immobility time is reduced by various antidepressants.

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

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

[0120] The embodiments of this disclosure described above are intended to be illustrative only. This disclosure may be embodied in other specific forms. Modifications, alterations, and variations of this disclosure may be made without departing from the intended scope. The systems, devices, and processes disclosed and shown herein may include a specific number of elements / components, but the systems, devices, and assemblies may be modified to include additional or fewer such elements / components. For example, any of the disclosed elements / components may be referred to as singular, but the embodiments disclosed herein may be modified to include multiple such elements / components. Features selected from one or more of the embodiments described above may be combined to create alternative embodiments not expressly described. All values ​​and sub-scopes within the scope of disclosure are also disclosed. The subject matter described herein is intended to encompass and include all preferred modifications in the art. All references mentioned herein are incorporated herein in their entirety by reference.

Claims

1. formula 【Chemistry 1】 Compounds of or pharmaceutically acceptable salts thereof.

2. formula 【Chemistry 2】 Compounds of or pharmaceutically acceptable salts thereof.

3. formula 【Transformation 3】 Compounds of or pharmaceutically acceptable salts thereof.

4. A method for treating depression, wherein the method is a formula 【Chemistry 4】 A method comprising administration of one or more compounds and pharmaceutically acceptable salts thereof.

5. A method for treating cancer, wherein the method is a formula 【Transformation 5】 A method comprising administration of one or more compounds and pharmaceutically acceptable salts thereof.

6. A method for treating one or more of the following conditions: depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's disease, and Parkinson's disease, wherein the method is a formula 【Transformation 6】 A method comprising administration of one or more compounds and pharmaceutically acceptable salts thereof.

7. The method according to claims 4 to 6, wherein the administration is performed orally.

8. The method according to claims 4 to 6, wherein the administration is by injection.

9. The method according to claims 4 to 8, wherein the administration is accompanied by a pharmaceutically acceptable additive.

10. A pharmaceutical composition for treating depression, wherein the composition is of the formula 【Transformation 7】 A pharmaceutical composition comprising one or more compounds and pharmaceutically acceptable salts thereof.

11. formula 【Transformation 8】 Use of the compound or pharmaceutically acceptable salt thereof for the treatment of depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's disease, or Parkinson's disease.

12. A pharmaceutical composition for treating depression, anxiety, multiple sclerosis, NASH, scleroderma, ADHD, Alzheimer's disease, or Parkinson's disease, wherein the composition is of the formula 【Chemistry 9】 A pharmaceutical composition comprising one or more compounds and pharmaceutically acceptable salts thereof.

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