Selective antagonists of the GluN2B subunit of the N-methyl-D-aspartate receptor with enhanced potency at acidic pH

Abstract

Disclosed are compounds that selectively inhibit GluN2B-containing N-methyl-D-aspartate receptors (NMDARs). In some cases, the compounds selectively target GluN2B over GluN2A, GluN2C, and / or GluN2D. In general, the compounds have enhanced potency against GluN2B at more acidic pH compared to physiological pH. Also disclosed are pharmaceutical formulations that contain one or more of the compounds. In addition, disclosed are methods of treating conditions, disorders, or diseases using the compounds or their pharmaceutical formulations.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 240,125, filed September 2, 2021, the entire contents of which are incorporated herein by reference.

[0002] Technical Field The present disclosure relates to N-methyl-D-aspartate receptor (NMDAR) modulators, in particular, to GluN2B subunit-selective allosteric modulators of NMDAR that have enhanced potency for GluN2B at acidic pH higher than physiological pH.The present disclosure also relates to pharmaceutical preparations containing such NMDAR modulators, and methods for treating conditions, disorders, and diseases using such NMDAR modulators. [Background technology]

[0003] background Cerebral ischemia, stroke, subarachnoid hemorrhage (SAH), and traumatic brain injury (TBI) all result in substantial neuronal death, and even if they are not fatal, they can result in long-term disability with significant social impact. Aside from clot lysis in a subset of patients or clot removal when occlusion occurs in the aorta, few therapeutic options are currently available for stroke. SAH can be treated with calcium channel blockers, but a significant proportion of patients progress to subsequent ischemic episodes and death, leaving significant opportunities for improved treatment. There are currently no approved pharmacological strategies for neuroprotection in TBI.

[0004] Extracellular glutamate concentrations are elevated in injured CNS tissues of animal models and human patients with acute injury (see Appendix Table S1 in Yuan, et al., Neuron, 2015, 85(6):1305-1318). One consequence of increased extracellular glutamate is overactivation of NMDARs, which can be neurotoxic (Choi, et al., J Neurosci, 1988, 8:185-196). Logically, inhibiting NMDARs during stress that elevates glutamate should provide neuroprotection, and the efficacy of several NMDAR antagonists has been confirmed in animal models of injury. However, promising preclinical results have yet to translate into clinical success, as multiple clinical trials using NMDAR antagonists for stroke or TBI have either failed to improve patient outcomes or have been associated with unacceptable side effects (Yuan, et al., Neuron, 2015, 85(6):1305-1318). Since the discovery of GluN2B-selective antagonists, various scaffolds of highly selective GluN2B-selective antagonists have been reported, and they have been shown to be effective in treating stroke (Yuan, et al., Neuron, 2015, 85(6):1305-1318), TBI (Yurkewicz, et al., J Neurotrauma, 2005, 22:1428-1443), Parkinson's disease (Michel, et al., PLoS One, 2014, 9(12):e114086, Michel, et al., PLoS One, 2015,10(8):e0135949), depression (Bristow, et al., J Pharmacol Exp Ther, 2017, 363(3):377-93), and pain (Swartjes, et al., Anesthesiology, 2011). 115(1):165-74; Labas, et al., Eur J Med Chem, 2011, 46(6):2295-309) have been tested in preclinical and clinical studies. Despite achieving apparent preclinical efficacy, no GluN2B-selective inhibitors have been approved for clinical use.

[0005] In some CNS conditions, disorders, and diseases, pH plays an important role in physiology. Neuronal action potential firing consumes energy by using ion gradients, which is associated with the movement of multiple organic and inorganic ions across the cell membrane. High firing rates of neurons are known to change extracellular pH (Kraig et al., J Neurophysiol, 1983, 49(3):831-50; Sykova et al., Ciba Found Symp, 1988, 139:220-35; Tong and Chesler, Brain Res., 1999, 815(2):373-81), and high-frequency firing releases substantial proton load (Theparambil, et al., Nat Commun, 2020, 11(1):5073). These protons are buffered by extracellular bicarbonate ions, but when firing rates increase or increased extracellular potassium is released (Kraig et al., J Neurophysiol, 1983, 49(3):831-50), as occurs during stroke, ischemia, hypoxia, and TBI (e.g., Mutch and Hansen, J Cereb Blood Flow Metab, 1984, 4(1):17-27), compensatory mechanisms to boost buffering capacity fail, resulting in substantial acidification (Theparambil, et al., Nat Commun, 2020, 11(1):5073). Repetitive stimulation of small primary afferent pain fibers can result in a progressive increase in action potential discharge, often referred to as windup (Woolf and Thompson, Pain, 1991, 44(3):293-299), and a sustained increase in spinal cord neuronal excitability. Such conditions, which result in high levels of action potential firing along pain pathways, are predicted to result in proton transfer to the extracellular space, as previously described. In some pathological conditions, such as chronic pain, firing rates can become substantial, resulting in local acidification, which sensitizes NMDARs to inhibitors with increased potency at low pH. In summary, there is an urgent need for GluN2B-selective NMDAR antagonists with improved preclinical and / or clinical outcomes, particularly for CNS conditions, disorders, and diseases. Furthermore, there is an urgent need for GluN2B-selective NMDAR antagonists with enhanced potency against GluN2B at acidic pH (higher than physiological pH). [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Yuan, et al., Neuron, 2015, 85(6):1305-1318 [Non-patent document 2] Choi, et al., J Neurosci, 1988, 8:185-196 [Non-patent document 3] Yurkewicz, et al., J Neurotrauma, 2005, 22:1428-1443 [Non-patent document 4] Michel, et al., PLoS One, 2014, 9(12):e114086 [Non-Patent Document 5] Michel, et al., PLoS One, 2015,10(8):e0135949 [Non-patent document 6] Swartjes, et al., Anesthesiology, 2011 115(1):165-74 [Non-Patent Document 7] Labas, et al., Eur J Med Chem, 2011, 46(6):2295-309 [Non-patent document 8] Kraig et al., J Neurophysiol, 1983, 49(3):831-50 [Non-Patent Document 9] Sykova et al., Ciba Found Symp, 1988, 139:220-35 [Non-Patent Document 10] Tong and Chesler, Brain Res., 1999, 815(2):373-81 [Non-Patent Document 11] Theparambil, et al., Nat Commun, 2020, 11(1):5073 [Non-Patent Document 12] Mutch and Hansen, J Cereb Blood Flow Metab, 1984, 4(1):17-27 [Non-Patent Document 13] Theparambil, et al., Nat Commun, 2020, 11(1):5073 [Non-Patent Document 14] Woolf and Thompson, Pain, 1991, 44(3):293-299 Summary of the Invention [Means for solving the problem]

[0007] overview The present disclosure describes a negative allosteric modulator that selectively inhibits NMDARs containing GluN2B subunits.In some cases, the negative allosteric modulator selectively targets GluN2B over GluN2A, GluN2C, and / or GluN2D.Generally, the negative allosteric modulator has enhanced potency against GluN2B at more acidic pH compared to physiological pH.

[0008] In some embodiments, the compounds disclosed herein have the structure of Formula I, or a pharmaceutically acceptable salt, hydrate, or hydrate salt of Formula I: [ka] [In the formula, R 1 teeth, [ka] Selected from R A , R B , R C , and R D are independently selected from hydrogen, methyl, and halomethyl; R 2 and R 3 are independently selected from hydrogen, methyl, and halomethyl.

[0009] In some embodiments, R 1 teeth, [ka] is.

[0010] In some embodiments, R 1 teeth, [ka] is.

[0011] In some embodiments, R 2 and R 3 are both hydrogen.

[0012] Exemplary compounds include: [ka] and their corresponding pharmaceutically acceptable salts, hydrates and hydrated salts.

[0013] In some embodiments, the compounds disclosed herein have the structure of Formula II, or a pharmaceutically acceptable salt, hydrate, or hydrate salt of Formula II: [ka] [In the formula, R 4 is selected from hydrogen, methyl, halomethyl, ethyl, haloethyl, isopropyl, and haloisopropyl; R 5 and R 6are independently selected from hydrogen, methyl, and halomethyl.

[0014] In some embodiments, R 4 is selected from methyl and halomethyl.

[0015] In some embodiments, R 5 and R 6 are both hydrogen.

[0016] Also disclosed are compositions containing the compounds described herein, wherein the compounds are present in greater than 80%, greater than 85%, greater than 90%, or greater than 95% enantiomeric excess relative to the stereocenter labeled with an "*" in the corresponding formulae disclosed herein. In some embodiments, the compounds in the compositions are present in greater than 95% enantiomeric excess relative to the stereocenter labeled with an "*" in the corresponding formulae disclosed herein.

[0017] In some embodiments, the composition contains a compound having the structure of Formula I, or a pharmaceutically acceptable salt, hydrate, or hydrated salt of Formula I, wherein the compound is present in an enantiomeric excess of greater than 80%, greater than 85%, greater than 90%, or greater than 95% in the R configuration with respect to the stereocenter labeled with an * as depicted in Formula I.

[0018] In some embodiments, the composition contains a compound having the structure of Formula II, or a pharmaceutically acceptable salt, hydrate, or hydrated salt of Formula II, wherein the compound is present in an enantiomeric excess of greater than 80%, greater than 85%, greater than 90%, or greater than 95% in the R configuration with respect to the stereocenter labeled with an * as depicted in Formula II.

[0019] Also disclosed are pharmaceutical formulations of the disclosed compounds or compositions. Typically, pharmaceutical formulations also contain pharmaceutically acceptable excipients. In some embodiments, the pharmaceutical formulation is in a form selected from tablets, capsules, caplets, pills, beads, granules, fine granules, powders, gels, creams, liquids, suspensions, emulsions, and nanoparticle formulations. In some embodiments, the pharmaceutical formulation is an oral formulation. In some embodiments, the pharmaceutical formulation is an intravenous formulation. In some embodiments, the pharmaceutical formulation is in the form of a lyophilized powder. In some embodiments, the pharmaceutical formulation is in the form of a sterile aqueous solution.

[0020] The present disclosure also relates to (1) the compounds, compositions, and pharmaceutical formulations disclosed herein for the treatment of a condition, disorder, or disease disclosed herein or for use as a medicament, (2) the compounds, compositions, and pharmaceutical formulations disclosed herein for use in the treatment of a condition, disorder, or disease disclosed herein, or (3) the compounds, compositions, and pharmaceutical formulations disclosed herein for the manufacture of a medicament for the treatment of a condition, disorder, or disease disclosed herein.

[0021] The present disclosure also provides a method for treating a condition, disorder, or disease in a subject in need thereof. The method comprises administering to the subject an effective amount of the compounds, compositions, and pharmaceutical formulations disclosed herein. In some embodiments, the compounds, compositions, or pharmaceutical formulations are administered orally or intravenously.

[0022] Exemplary conditions, disorders, and diseases relevant to the present disclosure include, but are not limited to, stroke, subarachnoid hemorrhage, cerebral ischemia, cerebral vasospasm, hypoxia, acute CNS injury, spinal cord injury, traumatic brain injury, coronary artery bypass graft, persistent or chronic cough, substance abuse disorders, opiate withdrawal, opiate tolerance, bipolar disorder, suicidal ideation, pain, fibromyalgia, depression, postpartum depression, resting tremor, dementia, epilepsy, seizure disorders, movement disorders, and neurodegenerative diseases.

[0023] In some embodiments, the condition, disorder or disease is pain, depression, stroke, or subarachnoid hemorrhage. [Brief explanation of the drawings]

[0024] [Figure 1] Figure 1 is a graph showing infarct volume (mm3) plotted against IP dose (mg / kg) of an exemplary compound (NP10679) in the MCAO model of transient ischemia in mice. Plots represent pooled data from three independent experiments. Data are shown as mean ± SEM for n = 9 (0.2 mg / kg), 13 (0.5 mg / kg), 21 (1 mg / kg), 12 (2 mg / kg), 12 (5 mg / kg), 24 (10 mg / kg), and 34 (Veh) mice. **p<0.01 from vehicle control (ANOVA, Dunnett).

[0025] [Figure 2A] Figure 2A is a graph showing the total plasma levels (ng / mL) of an exemplary compound (NP10679) plotted against time (hours) after oral administration of 10 mg / kg (black symbols) or IV administration of 3 mg / kg (white symbols) in mice. Data are presented as mean ± SEM (n=3 per data point).

[0026] [Figure 2B] Figure 2B is a graph showing the free plasma levels (nM) of an exemplary compound (NP10679) plotted against time (hours) after IP administration of 2 mg / kg (open symbols) or 5 mg / kg (closed symbols) in mice. Data are shown as mean ± SEM (n = 3 per data point). The IC50 of NP10679 for GluN2B at pH 6.9, the functional IC50 for the H1 histamine receptor, and the functional IC50 for hERG are shown as dotted lines on the graph.

[0027] [Figure 3]Figure 3 is a graph showing the latency (seconds) of mice to fall on the rotarod plotted against time (days). Mice were trained on the rotarod for two consecutive days (days 1 and 2), with four trials per day, with a 25-minute interval between trials. On day 3, mice were randomized into groups and administered vehicle control (open circle), 30 mg / kg of ifenprodil (open inverted triangle), or 2 mg / kg (open triangle), 5 mg / kg (black inverted triangle), or 10 mg / kg (cross) of an exemplary compound (NP10679). The latency to fall was calculated for each group and is shown as the mean ± SEM (n = 8). *p < 0.01 from vehicle control for individual trials on day 3 (ANOVA, Dunnett).

[0028] [Figure 4] Figure 4 is a bar graph showing the horizontal activity (within 2 hours) of mice among the three treatment groups: vehicle control, MK-801, and an exemplary compound (NP10679). Mice were habituated for 1 hour in a closed locomotor activity box, then removed and administered vehicle (Veh, n = 6), MK-801 (0.3 mg / kg, n = 4), or NP10679 (20 mg / kg, n = 6) by IP injection, and then placed back into the box. Horizontal locomotor activity was measured for 2 hours. *p<0.01 from vehicle control (ANOVA, Dunnett). The total number of beam breaks, representing horizontal movement during the sample period, is reported on the abscissa.

[0029] [Figure 5] Figures 5 and 6 are graphs showing the plasma exposure of an exemplary compound (NP10679) after a single intravenous administration in human subjects. Plasma collection and quantification were performed as described herein. Data expressed in ng / mL represent the average of 6 subjects per dose, except for the 150 mg group, which was the average of 5 subjects. [Figure 6]Figures 5 and 6 are graphs showing the plasma exposure of an exemplary compound (NP10679) after a single intravenous administration in human subjects. Plasma collection and quantification were performed as described herein. Data expressed in ng / mL represent the average of 6 subjects per dose, except for the 150 mg group, which was the average of 5 subjects. DETAILED DESCRIPTION OF THE INVENTION

[0030] Detailed Description The present disclosure describes a negative allosteric modulator that selectively inhibits NMDAR that contains GluN2B subunit.In some embodiments, the negative allosteric modulator selectively targets GluN2B over GluN2A, GluN2C, and / or GluN2D.Generally, the negative allosteric modulator has enhanced potency against GluN2B at more acidic pH compared to physiological pH.

[0031] Before describing the present disclosure in more detail, it should be understood that the present disclosure is not limited to particular embodiments described herein, and as such may, of course, vary according to the scope of the present disclosure. 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.

[0032] All publications and patents cited herein are incorporated by reference to the same extent as if each individual publication or patent was specifically and individually indicated to be incorporated by reference, and are incorporated by reference to disclose and describe the methods and / or materials in connection with which the publications and patents are cited.

[0033] As will be apparent to those skilled in the art upon reading this disclosure, each of the specific embodiments described and illustrated herein has distinct components and / or features which can be readily separated from or combined with one or more components and / or features of any of the other embodiments described herein without departing from the scope or spirit of the disclosure. Any recited method can be carried out in the order of events recited herein or in any other order which is logically possible.

[0034] Embodiments of the present disclosure employ, unless otherwise indicated, techniques of medicine, organic chemistry, medicinal chemistry, biochemistry, molecular biology, pharmacology, neurology, and the like, which are within the skill of the art and are fully explained in the literature, such as those cited herein.

[0035] I. Definition

[0036] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0037] As used herein, the term "subject" refers to animals, including humans and non-human animals. Non-human animals can include domestic pets, livestock and farm animals, and zoo animals. In some cases, non-human animals can be non-human primates.

[0038] As used herein, the terms "prevent" and "preventing" include prevention of occurrence, onset, spread, and / or recurrence. The present disclosure is not intended to be limited to complete prevention. For example, prevention is considered to be achieved if occurrence is delayed, the severity of onset is reduced, or both.

[0039] As used herein, the terms "treat" and "treating" include the medical management of a subject's condition, disorder, or disease, as would be understood by a person skilled in the art (see, for example, Stedman's Medical Dictionary). Generally, treatment is not limited to cases where the subject is cured and the condition, disorder, or disease is eradicated. Rather, treatment also contemplates cases where a treatment regimen containing one of the disclosed compounds, compositions, or pharmaceutical preparations results in an improved clinical outcome. Improved clinical outcomes may include one or more of the following: attenuation, reduction, and / or alleviation of one or more symptoms resulting from or associated with the condition, disorder or disease being treated; reduction in the occurrence of one or more symptoms; improvement in quality of life; reduction in the severity of the condition, disorder or disease; achieving or achieving a stabilized state of the condition, disorder or disease (i.e., not worsening); delay or slowing of the progression of the condition, disorder or disease; remission or alleviation of the condition, disorder or disease state; partial or total relief (whether detectable or undetectable); and improved survival (whether overall survival is prolonged or survival is prolonged compared to the expected survival of the subject if not treated). For example, the present disclosure encompasses treatments that reduce one or more symptoms of the neurological condition, disorder or disease described herein and / or cognitive impairment associated therewith.

[0040] As used herein, the term "physiological pH" refers to the pH normally found in the human body in the absence of pathological conditions. Typically, physiological pH ranges between 7.35 and 7.45, with an average of 7.40.

[0041] As used herein, the terms "halogen" and "halo" refer to fluorine, chlorine, bromine, and iodine.

[0042] As used herein, the term "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or formulations that are suitable, within the scope of sound medical judgment, for use in contact with the tissues of human and non-human animals without undue toxicity, irritation, allergic response, or other problem or complication, in accordance with the guidelines of a regulatory agency in a particular country, e.g., the Food and Drug Administration (FDA) in the United States, or its counterpart in a country outside the United States (e.g., the European Medicines Agency (EMA)).

[0043] As used herein, the term "salt" refers to an acid or base salt of the original compound. In some cases, salts are formed in situ during the preparation of the original compound, i.e., a salt is produced in place of the original compound by a specified synthetic chemical procedure. In some cases, salts are obtained by modifying the original compound. In some cases, salts are obtained by ion exchange with an existing salt of the original compound. Examples of salts include, but are not limited to, inorganic or organic acid salts of basic residues, such as amines, and alkali or organic salts of acidic residues, such as carboxylic acids and phosphoric acids. For original compounds containing basic residues, salts can be prepared by treating the compound with an appropriate amount of a non-toxic inorganic or organic acid; alternatively, salts can be formed in situ during the preparation of the original compound. Exemplary salts of basic residues include salts with inorganic acids selected from hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, and nitric acid, or salts with organic acids selected from acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, naphthalenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, and isethionic acid. For parent compounds containing acidic residues, salts can be prepared by treating the compound with an appropriate amount of a non-toxic base; alternatively, salts can be formed in situ during the preparation of the parent compound. Exemplary salts of acidic residues include salts with bases selected from ammonium hydroxide, sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, magnesium hydroxide, ferrous hydroxide, zinc hydroxide, copper hydroxide, aluminum hydroxide, ferric hydroxide, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, lysine, arginine, and histidine.If desired, salts can be prepared by reacting the free acid or base form of the parent compound with a stoichiometric or greater amount of an appropriate base or acid in water (including aqueous solutions), an organic solvent (including organic solutions), or a mixture thereof. Lists of exemplary pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 20th Ed., Lippincott Williams & Wilkins, Baltimore, MD, 2000, and Handbook of Pharmaceutical Salts: Properties, Selection, and Use, Stahl and Wermuth, Eds., Wiley-VCH, Weinheim, 2002.

[0044] As used herein, the term "excipient" refers to any component present in a pharmaceutical formulation disclosed herein other than the active ingredient (i.e., a compound or composition of the present disclosure).

[0045] As used herein, the term "effective amount" of a material refers to a non-toxic amount of a material sufficient to produce a desired result. The exact amount required may vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the condition, disorder, or disease being treated, the active ingredient or therapy used, etc.

[0046] II. Compounds

[0047] The present disclosure describes negative allosteric modulators that selectively inhibit GluN2B-containing NMDARs. In some embodiments, the negative allosteric modulators selectively target the GluN2B subunit of NMDARs over the GluN2A, GluN2C, and / or GluN2D subunits.

[0048] In some embodiments, the potency of the negative allosteric modulator for GluN2B increases as the environmental pH decreases to a pH range of 5.0-9.0, 6.0-8.0, 6.5-8.0, or 6.9-7.6. For example, the negative allosteric modulator has enhanced potency for GluN2B at more acidic pH compared to physiological pH. Potency for GluN2B is measured by the IC of the negative allosteric modulator for GluN2B. 50 It can be evaluated by its IC 50 The IC value can be easily determined by the method described in the Examples. 50 Lower values ​​correspond to higher potency.

[0049] To the extent that the chemical formulas described herein contain one or more unspecified chiral centers, the formulas are intended to encompass all stable stereoisomers, enantiomers, and diastereomers. Such compounds can exist as single enantiomers, racemic mixtures, diastereomeric mixtures, or combinations thereof. It is also understood that the chemical formulas encompass all tautomeric forms, where tautomers are possible.

[0050] Methods for making exemplary compounds are disclosed in the Examples. The methods are compatible with a wide variety of functional groups and compounds, and therefore a wide variety of derivatives can be obtained from the disclosed methods.

[0051] A. General structure

[0052] Formula I In some embodiments, the compound has the structure of Formula I, or a pharmaceutically acceptable salt, hydrate, or hydrate salt of Formula I: [ka] [In the formula, R 1 teeth, [ka] Selected from RA , R B , R C , and R D are independently selected from hydrogen, methyl, and halomethyl (e.g., fluoromethyl, e.g., mono-, di-, and trifluoromethyl); R 2 and R 3 are independently selected from hydrogen, methyl, and halomethyl (e.g., fluoromethyl, e.g., mono-, di-, and trifluoromethyl).

[0053] In some embodiments, the compound is in the free base form shown in Formula I. In some embodiments, the compound is a pharmaceutically acceptable salt of Formula I.

[0054] In some embodiments, R 1 teeth, [ka] is.

[0055] In some embodiments, R 1 teeth, [ka] is.

[0056] In some embodiments, R 1 teeth, [ka] is.

[0057] In some embodiments, R 1 teeth, [ka] is.

[0058] In some embodiments, R 1 teeth, [ka] is. In some embodiments, R 1 teeth,

[0059] [ka] is.

[0060] In some embodiments, R 1 teeth, [ka] is.

[0061] In some embodiments, R 1 teeth, [ka] is.

[0062] In some embodiments, R 1 teeth, [ka] is.

[0063] In some embodiments, R 1 teeth, [ka] is.

[0064] In some embodiments, R 1 teeth, [ka] is.

[0065] In some embodiments, R 1 teeth, [ka] is.

[0066] In some embodiments, R 1 teeth, [ka] is.

[0067] In some embodiments, R A is hydrogen. In some embodiments, R B is hydrogen. In some embodiments, R A and R B are hydrogen atoms.

[0068] In some embodiments, R C is hydrogen. In some embodiments, R D is hydrogen. In some embodiments, R C and R D are hydrogen atoms.

[0069] In some embodiments, R 2 is hydrogen. In some embodiments, R 2 is methyl. In some embodiments, R 2 is halomethyl, for example, fluoromethyl, for example, mono-, di-, and trifluoromethyl.

[0070] In some embodiments, R 3 is hydrogen. In some embodiments, R 3 is methyl. In some embodiments, R 3 is halomethyl, for example, fluoromethyl, for example, mono-, di-, and trifluoromethyl.

[0071] In some embodiments, R 2 and R 3 are each hydrogen. 2 is hydrogen and R 3 is methyl or halomethyl. In some embodiments, R 2 is methyl or halomethyl, and R 3 is hydrogen. In some embodiments, R 2and R 3 is independently methyl or halomethyl.

[0072] In some embodiments, the compound has the structure of formula IA, or a pharmaceutically acceptable salt, hydrate, or hydrate salt of formula IA: [ka] [In the formula, R 1 are the same as those described above for Formula I].

[0073] Exemplary compounds include: [ka] and their corresponding pharmaceutically acceptable salts, hydrates and hydrated salts.

[0074] Formula II In some embodiments, the compound has the structure of Formula II, or a pharmaceutically acceptable salt, hydrate, or hydrate salt of Formula II: [ka] [In the formula, R 4 is selected from hydrogen, methyl, halomethyl (e.g., fluoromethyl, for example, mono-, di-, and trifluoromethyl), ethyl, haloethyl (e.g., fluoroethyl, for example, mono-, di-, and trifluoroethyl), isopropyl, and haloisopropyl (e.g., fluoroisopropyl, for example, mono-, di-, and trifluoroisopropyl); R 5 and R 6 are independently selected from hydrogen, methyl, and halomethyl (e.g., fluoromethyl, e.g., mono-, di-, and trifluoromethyl).

[0075] In some embodiments, the compound is in the free base form shown in Formula II: In some embodiments, the compound is a pharmaceutically acceptable salt of Formula II.

[0076] In some embodiments, R 4 is hydrogen. In some embodiments, R 4 is methyl. In some embodiments, R 4 is halomethyl, e.g., fluoromethyl, e.g., mono-, di-, and trifluoromethyl. In some embodiments, R 4 is ethyl. In some embodiments, R 4 is haloethyl, e.g., fluoroethyl, e.g., mono-, di-, and trifluoroethyl. In some embodiments, R 4 is isopropyl. In some embodiments, R 4 is haloisopropyl, for example, fluoroisopropyl, for example, mono-, di-, and trifluoroisopropyl.

[0077] In some embodiments, R 4 is selected from methyl and halomethyl (eg, fluoromethyl, eg, mono-, di-, and trifluoromethyl).

[0078] In some embodiments, R 5 is hydrogen. In some embodiments, R 5 is methyl. In some embodiments, R 5 is halomethyl, for example, fluoromethyl, for example, mono-, di-, and trifluoromethyl.

[0079] In some embodiments, R 6 is hydrogen. In some embodiments, R 6 is methyl. In some embodiments, R 6 is halomethyl, for example, fluoromethyl, for example, mono-, di-, and trifluoromethyl.

[0080] In some embodiments, R 5 and R 6 are each hydrogen. 5 is hydrogen and R 6 is methyl or halomethyl. In some embodiments, R 5is methyl or halomethyl, and R 6 is hydrogen. In some embodiments, R 5 and R 6 is independently methyl or halomethyl.

[0081] In some embodiments, the compound has the structure of Formula IIA, or a pharmaceutically acceptable salt, hydrate, or hydrate salt of Formula IIA: [ka] [In the formula, R 4 is the same as previously described for Formula II].

[0082] B. Stereochemistry and pH Sensitivity

[0083] The foregoing compounds exist in the R configuration with respect to the chiral centers labeled with an "*" symbol in the formula.

[0084] In certain embodiments, compounds have higher potency at GluN2B than their corresponding S enantiomers. Potency at GluN2B is determined by the IC of the compound at GluN2B. 50 This IC can be evaluated by the value 50 The IC value can be easily determined by the method described in the Examples. 50 Lower values ​​correspond to higher potency.

[0085] In some embodiments, the potency of a compound against GluN2B increases when the environmental pH is decreased to a pH range of 5.0-9.0, 6.0-8.0, 6.5-8.0, or 6.9-7.6. For example, a compound has enhanced potency against GluN2B at a more acidic pH compared to physiological pH. Here, the IC determined at pH 6.9 for a particular compound is 50 IC values ​​determined at pH 7.6 50 The ratio of the values ​​is defined as the "pH boost" of the compound.

[0086] In some embodiments, the compounds have a pH boost that is equal to or greater than that of their corresponding S enantiomers. As used herein, "equivalent" refers to a value within 25% variation of the comparative value. In some embodiments, the compounds have a pH boost that is equal to or greater than 75% of that of their corresponding S enantiomers. In some embodiments, the compounds have a pH boost that is equal to or greater than 80% of that of their corresponding S enantiomers. In some embodiments, the compounds have a pH boost that is equal to or greater than 85% of that of their corresponding S enantiomers. In some embodiments, the compounds have a pH boost that is equal to or greater than 90% of that of their corresponding S enantiomers. In some embodiments, the compounds have a pH boost that is equal to or greater than 95% of that of their corresponding S enantiomers.

[0087] III. Composition

[0088] Disclosed herein is a composition comprising the compound disclosed herein. In some embodiments, the compound in the composition is present with an enantiomeric excess of more than 80%, more than 85%, more than 90%, or more than 95% with respect to the stereocenter marked with "*" in any one of formulas I, IA, II, and IIA. In some embodiments, the compound in the composition is present with an enantiomeric excess of more than 95% with respect to the stereocenter marked with "*" in any one of formulas I, IA, II, and IIA.

[0089] In some embodiments, the composition contains a compound having the structure of Formula I, or a pharmaceutically acceptable salt, hydrate, or hydrated salt of Formula I, wherein the compound in the composition is present in an enantiomeric excess of greater than 80%, greater than 85%, greater than 90%, or greater than 95% of the R configuration as depicted by Formula I with respect to the stereocenter labeled with an *. In some embodiments, the compound in the composition is present in an enantiomeric excess of greater than 95% of the R configuration as depicted by Formula I with respect to the stereocenter labeled with an *.

[0090] In some embodiments, the composition contains a compound having the structure of formula IA, or a pharmaceutically acceptable salt, hydrate, or hydrated salt of formula IA, wherein the compound in the composition is present in an enantiomeric excess of greater than 80%, greater than 85%, greater than 90%, or greater than 95% of the R configuration as depicted by formula IA with respect to the stereocenter labeled with an *. In some embodiments, the compound in the composition is present in an enantiomeric excess of greater than 95% of the R configuration as depicted by formula IA with respect to the stereocenter labeled with an *.

[0091] In some embodiments, the composition contains a compound having the structure of Formula II, or a pharmaceutically acceptable salt, hydrate, or hydrated salt of Formula II, wherein the compound in the composition is present in an enantiomeric excess of greater than 80%, greater than 85%, greater than 90%, or greater than 95% of the R configuration as depicted by Formula II with respect to the stereocenter labeled with an *. In some embodiments, the compound in the composition is present in an enantiomeric excess of greater than 95% of the R configuration as depicted by Formula II with respect to the stereocenter labeled with an *.

[0092] In some embodiments, the composition comprises a compound having the structure of Formula IIA, or a pharmaceutically acceptable salt, hydrate, or hydrated salt of Formula IIA, wherein the compound in the composition is present in an enantiomeric excess of greater than 80%, greater than 85%, greater than 90%, or greater than 95% of the R configuration as depicted by Formula IIA with respect to the stereocenter labeled with an *. In some embodiments, the compound in the composition is present in an enantiomeric excess of greater than 95% of the R configuration as depicted by Formula IIA with respect to the stereocenter labeled with an *.

[0093] The disclosed compounds may exist in a mixture of salt and non-salt forms. In some embodiments, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 95%, or more than 98% of the compounds in the mixture may be in non-salt form, calculated as the ratio of the weight of the non-salt form to the total weight of the salt and non-salt forms. In some embodiments, more than 90% of the compounds in the mixture may be in non-salt form. In some embodiments, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 95%, or more than 98% of the compounds in the mixture may be in salt form, calculated as the ratio of the weight of the salt form to the total weight of the salt and non-salt forms. In some embodiments, more than 90% of the compounds in the mixture may be in salt form.

[0094] IV. Formulations

[0095] Pharmaceutical formulations containing the compounds or compositions described herein are disclosed. Generally, pharmaceutical formulations also contain one or more pharmaceutically acceptable excipients.

[0096] The pharmaceutical formulation may be in a form selected from tablets, capsules, caplets, pills, powders, beads, granules, fine granules, creams, gels, liquids (e.g., aqueous solutions, e.g., saline and buffered saline), emulsions, suspensions (including nano- and microsuspensions), nanoparticle formulations, and the like. In some embodiments, the pharmaceutical formulation is an oral formulation. In some embodiments, the pharmaceutical formulation is an intravenous formulation. In some embodiments, the pharmaceutical formulation is a topical formulation.

[0097] In some embodiments, pharmaceutical preparations are in the form of lyophilized powder.In some embodiments, lyophilized powder is prepared by dissolving active ingredient (i.e., the compound or composition disclosed herein) in aqueous solution, followed by lyophilization.For example, lyophilized powder can be prepared by dissolving active ingredient in phosphate buffered hydroxy-beta cyclodextrin solution, followed by lyophilization.

[0098] In some embodiments, the pharmaceutical preparation is in the form of a sterile aqueous solution. In some embodiments, the sterile aqueous solution is sterile PBS. In some embodiments, the sterile aqueous solution is prepared by dissolving a lyophilized powder containing the active ingredient (i.e., the compound or composition disclosed herein) in an aqueous solution. For example, the sterile aqueous solution can be prepared by dissolving a lyophilized powder containing the active ingredient in a volume appropriate for dosage of sterile PBS. In some embodiments, the lyophilized powder containing the active ingredient is the same as that described in the above paragraph.

[0099] As used herein, "emulsion" refers to a mixture of immiscible components that are uniformly blended together. In some forms, the immiscible components include a lipophilic component and an aqueous component. For example, an emulsion can be a preparation in which one liquid is distributed in small globules throughout a second liquid. The dispersion is the discontinuous phase, and the dispersion medium is the continuous phase. When oil or oily substance is the dispersion and water or an aqueous solution is the continuous phase, it is known as an oil-in-water emulsion; when water or an aqueous solution is the dispersed phase and oil or oily substance is the continuous phase, it is known as a water-in-oil emulsion.

[0100] As used herein, "biocompatible" refers to materials that, in and of themselves, are not toxic to the host (e.g., non-human animals or humans) and do not degrade at a rate that produces toxic concentrations of monomeric or oligomeric subunits or other by-products in the host (if the material degrades).

[0101] As used herein, "biodegradable" refers to the breakdown or disintegration of a polymeric material into smaller (eg, non-polymeric) subunits, or the digestion of the material into smaller subunits.

[0102] As used herein, "enteric polymer" refers to a polymer that becomes soluble in the higher pH environment of the lower gastrointestinal tract or that slowly erodes as the dosage form passes through the gastrointestinal tract.

[0103] As used herein, a "nanoparticle formulation" generally refers to "nanoparticles," which are particles having a diameter of about 1 nm to 1000 nm, about 10 nm to 1000 nm, about 100 nm to 1000 nm, or about 250 nm to 1000 nm. In some embodiments, a "nanoparticle formulation" can also refer to "microparticles," which are particles having a diameter of about 1 micron to about 100 microns, about 1 to about 50 microns, about 1 to about 30 microns, or about 1 micron to about 10 microns. In some embodiments, a nanoparticle formulation can be a mixture of nanoparticles, as defined above, and microparticles, as defined above.

[0104] As used herein, "surfactant" refers to any agent that preferentially absorbs at the interface between two immiscible phases, such as the interface between water (or an aqueous solution) and an organic solvent (or an organic solution), the water / air interface, and the organic solvent / air interface. Surfactants generally have a hydrophilic portion and a lipophilic portion.

[0105] As used herein, a "gel" is a semi-solid system containing a dispersion of an active ingredient, i.e., a compound or composition according to the present disclosure, in a liquid vehicle that has been made semi-solid by the action of a thickening agent or polymeric material dissolved or suspended in the liquid vehicle. The liquid vehicle may contain a lipophilic component, an aqueous component, or both.

[0106] As used herein, "hydrogel" refers to a water-containing, swollen network of water-insoluble, finely dispersed polymer chains, in which the polymer molecules reside in the external or dispersed phase and water (or aqueous solution) forms the internal or dispersed phase. The polymer chains can be chemically crosslinked (chemical gels) or physically crosslinked (physical gels). Chemical gels have polymer chains that are joined through covalent bonds, while physical gels have polymer chains connected by non-covalent interactions, such as van der Waals interactions, ionic interactions, hydrogen-bonding interactions, and hydrophobic interactions.

[0107] As used herein, "beads" refers to beads made using an active ingredient (i.e., a compound or composition according to the present disclosure) and one or more pharmaceutically acceptable excipients. Beads can be produced by applying the active ingredient to an inert support, for example, by coating an inert sugar core with the active ingredient. Alternatively, beads can be produced by fabricating a "core" containing both the active ingredient and at least one of one or more pharmaceutically acceptable excipients. As used herein, "granules" refers to products made by processing particles of the active ingredient (i.e., a compound or composition according to the present disclosure), which may or may not contain one or more pharmaceutically acceptable excipients. Typically, granules do not contain an inert support and are larger in size than the particles used to produce granules. While beads, granules, and fine granules can be formulated to provide immediate release, beads and granules are usually used to provide delayed release.

[0108] As used herein, "enzymatically degradable polymer" refers to a polymer that is degraded by bacterial enzymes present in the intestine and / or lower gastrointestinal tract.

[0109] A. Physical Form and Unit Dosage

[0110] The compounds or compositions described herein can be formulated in various ways depending on the mode of introduction. Pharmaceutical preparations can be prepared in various forms, such as tablets, capsules, caplets, pills, granules, powders, nanoparticle preparations, liquids (e.g., aqueous solutions, such as saline and buffered saline), suspensions (including nano- and micro-suspensions), emulsions, creams, gels, etc.

[0111] In some embodiments, pharmaceutical preparations are present in a solid dosage form suitable for easy, preferably oral administration with precise dosage.Solid dosage forms for oral administration include, but are not limited to, tablets, soft or hard gelatin or non-gelatin capsules, and caplets.However, liquid dosage forms, such as solutions, suspensions (including nano- and micro-suspensions), and emulsions, can also be used.Intravenous preparations are usually present in liquid dosage forms, including solutions, emulsions, and suspensions.Suitable topical preparations include, but are not limited to, creams and gels.

[0112] In some embodiments, the pharmaceutical formulations are present in unit dosage form and may be suitably packaged, for example, in a box, blister, vial, bottle, sachet, ampoule, or any other suitable single-dose or multi-dose holder or container, optionally along with one or more leaflets containing product information and / or instructions for use.

[0113] In certain embodiments, the amount of a compound disclosed herein in a unit dose is an amount suitable for single daily administration. In certain embodiments, multiple unit doses are required to reach the desired total daily dose.

[0114] In certain embodiments, a unit dose can contain between 5 and 300 mg of a compound disclosed herein. In certain embodiments, the amount of a compound disclosed herein in a unit dose can be between about 5 and about 300 mg, about 15 and about 300 mg, about 25 and about 300 mg, about 50 and about 300 mg, about 75 and about 300 mg, about 5 and about 250 mg, about 15 and about 250 mg, about 25 and about 250 mg, about 50 and about 250 mg, about 75 and about 250 mg, about 5 and about 200 mg, about 15 and about 200 mg, or about 200 mg. mg, about 25 to about 200 mg, about 50 to about 200 mg, about 75 to about 200 mg, about 5 to about 175 mg, about 15 to about 175 mg, about 25 to about 175 mg, about 50 to about 175 mg, about 75 to about 175 mg, about 5 to about 150 mg, about 15 to about 150 mg, about 25 to about 150 mg, about 50 to about 150 mg, about 75 to about 150 mg, or about 100 to about 150 mg.

[0115] In some embodiments, the unit dosage contains between 5 and 200 mg of a compound disclosed herein.

[0116] In some embodiments, the unit dosage contains between 25 and 200 mg of a compound disclosed herein.

[0117] In some embodiments, the unit dose contains between 25 and 175 mg of a compound disclosed herein. In some embodiments, the unit dose contains between 25 and 150 mg of a compound disclosed herein. In some embodiments, the unit dose contains between 50 and 200 mg of a compound disclosed herein. In some embodiments, the unit dose contains between 75 and 200 mg of a compound disclosed herein. In some embodiments, the unit dose contains between 50 and 175 mg of a compound disclosed herein. In some embodiments, the unit dose contains between 75 and 150 mg of a compound disclosed herein.

[0118] In certain embodiments, the amount of the compound disclosed herein in unit dosage is about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, or about 300 mg.In a specific embodiment, the amount of the compound disclosed herein in unit dosage is about 100 mg.In a specific embodiment, the amount of the compound disclosed herein in unit dosage is about 150 mg.

[0119] Generally, the total daily dosage to be administered in one or more doses to a human subject in need thereof is between 5 and 300 mg of a compound disclosed herein. In certain embodiments, the total daily dosage of a compound disclosed herein is between about 5 and about 300 mg, about 15 and about 300 mg, about 25 and about 300 mg, about 50 and about 300 mg, about 75 and about 300 mg, about 5 and about 250 mg, about 15 and about 250 mg, about 25 and about 250 mg, about 50 and about 250 mg, about 75 and about 250 mg, about 5 and about 200 mg, or about 15 and about 200 mg. , about 25 to about 200 mg, about 50 to about 200 mg, about 75 to about 200 mg, about 5 to about 175 mg, about 15 to about 175 mg, about 25 to about 175 mg, about 50 to about 175 mg, about 75 to about 175 mg, about 5 to about 150 mg, about 15 to about 150 mg, about 25 to about 150 mg, about 50 to about 150 mg, about 75 to about 150 mg, or about 100 to about 150 mg.

[0120] Generally, the total daily dosage to be administered to a human subject in one or more doses is between about 11 and about 667 mmol of a compound disclosed herein. In certain embodiments, the total daily dosage of a compound disclosed herein is between about 11 and about 667 mmol, about 33 and about 667 mmol, about 56 and about 667 mmol, about 111 and about 667 mmol, about 167 and about 667 mmol, about 11 and about 556 mmol, about 33 and about 556 mmol, about 56 and about 556 mmol, about 111 and about 556 mmol, about 167 and about 556 mmol, about 11 and about 445 mmol, about 33 and about 445 mmol, about 5 and about 667 ... The range is from about 6 to about 445 mmol, from about 111 to about 445 mmol, from about 167 to about 445 mmol, from about 11 to about 389 mmol, from about 33 to about 389 mmol, from about 56 to about 389 mmol, from about 111 to about 389 mmol, from about 167 to about 389 mmol, from about 11 to about 334 mmol, from about 33 to about 334 mmol, from about 56 to about 334 mmol, from about 111 to about 334 mmol, from about 167 to about 334 mmol, or from about 222 to about 334 mmol.

[0121] In certain embodiments, the treatment course includes a loading dose per day for one or several days, followed by a reduced or normal dose per day for one or several days.For example, the treatment course can include a loading dose on the first day, followed by a reduced or normal dose per day for the remainder of the course.A suitable loading dose can be selected from the above-mentioned exemplary total daily doses.A suitable reduced or normal dose can also be selected from the above-mentioned exemplary total daily doses.In certain embodiments, the loading dose is about 150 mg, and the reduced or normal dose is 100 mg.For example, the treatment course can include a loading dose of 150 mg on the first day, followed by a reduced or normal dose of 100 mg per day for the remainder of the course.

[0122] B. Pharmaceutically Acceptable Excipients

[0123] Exemplary pharmaceutically acceptable excipients include, but are not limited to, diluents (fillers), binders, lubricants, disintegrants, pH correctors or buffering agents, preservatives, antioxidants, solubility enhancers, wetting agents or emulsifiers, plasticizers, colorants (e.g., pigments and dyes), flavoring or sweetening agents, thickeners, emollients, moisturizers, stabilizers, glidants, solvents or dispersion media, surfactants, pore formers, and coating or matrix materials.

[0124] In some embodiments, the tablets, beads, granules, and fine granules described herein contain one or more of the following pharmaceutically acceptable excipients: diluents, binders, lubricants, disintegrants, pigments, stabilizers, and surfactants. If desired, the tablets, beads, granules, and fine granules can also contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, dyes, pH buffering agents, and preservatives.

[0125] Examples of coating or matrix materials include, but are not limited to, cellulose polymers (e.g., methylcellulose, ethylcellulose, cellulose acetate, cellulose acetate phthalate, hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxymethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate, cellulose acetate trimellitate, and sodium carboxymethylcellulose), vinyl polymers and copolymers (e.g., polyvinylpyrrolidone, polyvinyl acetate, polyvinyl acetate phthalate, vinyl acetate-crotonic acid copolymer, and ethylene-vinyl acetate copolymer), acrylic acid polymers and copolymers (e.g., those formed from acrylic acid, methacrylic acid, methyl acrylate, ethyl acrylate, methyl methacrylate, ethyl methacrylate, and other methacrylic resins commercially available under the trademark EUDRAGIT®), enzymatically degradable polymers (e.g., azopolymers, pectin, chitosan, amylose, and guar gum), zein, shellac, and polysaccharides. In some embodiments, the coating or matrix material may contain one or more excipients, such as plasticizers, colorants, glidants, stabilizers, pore formers, and surfactants.

[0126] In some embodiments, the coating or matrix material is a pH-sensitive or pH-responsive polymer, such as the enteric polymers commercially available under the trademark EUDRAGIT®. For example, EUDRAGIT® L30D-55 and L100-55 are soluble at pH 5.5 and above, EUDRAGIT® L100 is soluble at pH 6.0 and above, and EUDRAGIT® S, as a result of its higher degree of esterification, is soluble at pH 7.0 and above.

[0127] In some embodiments, the coating or matrix material is a water-insoluble polymer with different degrees of permeability and extensibility, such as EUDRAGIT® NE, RL, and RS.

[0128] Depending on the coating or matrix material, disintegration / degradation or structural changes of the pharmaceutical formulation may occur at different locations in the gastrointestinal tract, hi some embodiments, the coating or matrix material is selected such that the pharmaceutical formulation can survive exposure to gastric acid after oral administration and release the active ingredient in the intestine.

[0129] Diluents, also called "fillers," can increase the bulk of a solid dosage formulation to provide a practical size for tablet compression or formation of beads, granules, or fine granules. Suitable diluents include, but are not limited to, dicalcium phosphate dihydrate, calcium sulfate, lactose, sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, kaolin, sodium chloride, dry starch, hydrolyzed starch, pregelatinized starch, silicon dioxide, titanium oxide, magnesium aluminum silicate, powdered sugar, and combinations thereof.

[0130] Binders impart cohesive properties to solid formulations and are therefore used to ensure that tablets, beads, granules, or fine granules remain intact after formation of the solid formulation. Suitable binder materials include, but are not limited to, starch, pregelatinized starch, gelatin, sugars (e.g., sucrose, glucose, dextrose, lactose, and sorbitol), polyethylene glycol, waxes, natural and synthetic gums (e.g., acacia, tragacanth, and sodium alginate), celluloses (e.g., hydroxypropylmethylcellulose, hydroxypropylcellulose, and ethylcellulose), veegum, and synthetic polymers (e.g., acrylic acid copolymers, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, polyacrylic acid, polymethacrylic acid, and polyvinylpyrrolidone), and combinations thereof.

[0131] Lubricants are used to facilitate tablet manufacture. Suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, glyceryl behenate, polyethylene glycol, talc, and mineral oil.

[0132] Disintegrants are used to facilitate the disintegration or "breakup" of a solid formulation after administration and generally include, but are not limited to, starch, sodium starch glycolate, sodium carboxymethyl starch, sodium carboxymethylcellulose, hydroxypropyl cellulose, pregelatinized starch, clays, cellulose, gums, and cross-linked polymers such as cross-linked polyvinylpyrrolidone (e.g., POLYPLASDONE® XL from GAF Chemical Corp.).

[0133] Plasticizers are usually present to create or promote plasticity and flexibility and reduce brittleness. Examples of plasticizers include polyethylene glycol, propylene glycol, triacetin, dimethyl phthalate, diethyl phthalate, dibutyl phthalate, dibutyl sebacate, triethyl citrate, tributyl citrate, acetyl triethyl citrate, castor oil, and acetylated monoglycerides.

[0134] Stabilizers are used to inhibit or delay the decomposition reaction of active ingredients in pharmaceutical formulations, or to stabilize particles in dispersions.For example, when the decomposition reaction involves the oxidation reaction of active ingredients in pharmaceutical formulations, stabilizers can be antioxidants or reducing agents.Stabilizers also include nonionic emulsifiers, such as sorbitan esters, polysorbates, and polyvinylpyrrolidone.

[0135] Glidants are used to reduce sticking effects during film formation and drying. Exemplary glidants include, but are not limited to, talc, magnesium stearate, and glyceryl monostearate.

[0136] Preservatives can inhibit deterioration and / or degradation of pharmaceutical formulations. Deterioration or degradation can be caused by one or more of microbial growth, fungal growth, and undesirable chemical or physical changes. Suitable preservatives include benzoates (e.g., sodium benzoate), ascorbic acid, methyl hydroxybenzoate, ethyl p-hydroxybenzoate, n-propyl p-hydroxybenzoate, n-butyl p-hydroxybenzoate, potassium sorbate, sorbic acid, propionates (e.g., sodium propionate), chlorobutanol, benzyl alcohol, and combinations thereof.

[0137] The surfactant may be anionic, cationic, amphoteric, or nonionic. Exemplary anionic surfactants include, but are not limited to, those containing carboxylate, sulfonate, or sulfate ions. Examples of anionic surfactants include sodium, potassium, and ammonium long-chain (e.g., 13-21) alkyl sulfonates (e.g., sodium lauryl sulfate), sodium, potassium, and ammonium aryl alkyl sulfonates (e.g., sodium dodecylbenzenesulfonate), and sodium dialkyl sulfosuccinates (e.g., sodium bis-(2-ethylthioxyl)-sulfosuccinate). Cationic surfactants include, but are not limited to, quaternary ammonium compounds such as benzalkonium chloride, benzethonium chloride, cetrimonium bromide, stearyldimethylbenzylammonium chloride, polyoxyethylene, and coconut amine. Examples of nonionic surfactants include ethylene glycol monostearate, propylene glycol myristate, glyceryl monostearate, glyceryl stearate, polyglyceryl-4-oleate, sorbitan acylate, sucrose acylate, PEG-150 laurate, PEG-400 monolaurate, polyoxyethylene monolaurate, polysorbate, polyoxyethylene octylphenyl ether, PEG-1000 cetyl ether, polyoxyethylene tridecyl ether, polypropylene glycol butyl ether, poloxamer (e.g., poloxamer 401), stearoyl monoisopropanolamide, and polyoxyethylene hydrogenated tallow amide. Examples of amphoteric surfactants include, but are not limited to, sodium N-dodecyl-β-alanine, sodium N-lauryl-β-iminodipropionate, myristoamphoacetate, lauryl betaine, and lauryl sulfobetaine.

[0138] Liquid pharmaceutical preparations typically contain solvents or dispersion media, such as water, aqueous solutions (e.g., saline, buffered saline, etc.), ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), oils (e.g., vegetable oils, such as peanut oil, corn oil, sesame oil, etc.), and combinations thereof.In some embodiments, liquid pharmaceutical preparations are aqueous preparations.Suitable solvents or dispersion media for intravenous preparations include, but are not limited to, water, saline, buffered saline (e.g., phosphate buffered saline), and Ringer's solution.

[0139] C. Pharmaceutically Acceptable Carriers

[0140] In some embodiments, pharmaceutical formulations are prepared using pharmaceutically acceptable carriers that encapsulate, embed, entrap, dissolve, disperse, absorb, and / or bind the compounds or compositions disclosed herein. Pharmaceutically acceptable carriers are composed of materials that are considered safe and can be administered to a subject without causing undesired biological side effects or undesired interactions. Preferably, pharmaceutically acceptable carriers do not interfere with the effectiveness of the compounds or compositions performing their functions. Pharmaceutically acceptable carriers can be formed from biodegradable materials, non-biodegradable materials, or a combination thereof. The aforementioned pharmaceutically acceptable excipients can be present partially or entirely in the pharmaceutically acceptable carrier.

[0141] In some embodiments, the pharmaceutically acceptable carrier is a controlled release carrier, for example, a delayed release carrier, a sustained release (extended release) carrier, and a pulsatile release carrier.

[0142] In some embodiments, the pharmaceutically acceptable carrier is pH sensitive or pH responsive.In some embodiments, the pharmaceutically acceptable carrier can be decomposed or degraded in a certain pH range.In some embodiments, the pharmaceutically acceptable carrier can undergo structural changes when subjected to pH changes.

[0143] Exemplary pharmaceutically acceptable carriers include, but are not limited to, nanoparticles, microparticles and combinations thereof, liposomes, hydrogels, polymer matrices, and solvent systems.

[0144] In some embodiments, the pharmaceutically acceptable carrier is a nanoparticle, a microparticle, or a combination thereof, hi some embodiments, the compound or composition is embedded in a matrix formed by the nanoparticle, microparticle, or combination thereof.

[0145] The nanoparticles, microparticles, or combinations thereof may be biodegradable and can be biodegraded at a controlled rate as needed for delivery of the compound or composition. The nanoparticles, microparticles, or combinations thereof may be made from a variety of materials. Both inorganic and organic materials may be used. Both polymeric and non-polymeric materials may be used.

[0146] For example, nanoparticles, microparticles, or combinations thereof are formed from one or more biocompatible polymers. In some embodiments, the biocompatible polymer is biodegradable. In some embodiments, the biocompatible polymer is non-biodegradable. In some embodiments, nanoparticles, microparticles, or combinations thereof are formed from a mixture of biodegradable and non-biodegradable polymers. The polymers used to form nanoparticles, microparticles, or combinations thereof can be adjusted to optimize various characteristics of the nanoparticles, microparticles, or combinations thereof, including (i) the interaction between the compound and the polymer to stabilize the compound and maintain activity during delivery, (ii) the rate of polymer degradation and therefore the release rate, (iii) the surface characteristics and targeting ability through chemical modification, and (iv) the porosity of the particle.

[0147] Exemplary polymers include polymers prepared from lactones, such as poly(caprolactone) (PCL), polyhydroxy acids and their copolymers, such as poly(lactic acid) (PLA), poly(glycolic acid) (PGA), poly(lactic-co-glycolic acid) (PLGA), and blends thereof, polyalkyl cyanoacrylates, polyurethanes, polyamino acids, such as poly-L-lysine (PLL), poly(valeric acid), and poly-L-glutamic acid, hydroxypropyl methacrylate, and the like. Polyvinyl ethers (HPMA), polyanhydrides, polyorthoesters, poly(ester amides), polyamides, poly(ester ethers), polycarbonates, ethylene vinyl acetate polymers (EVA), polyvinyl alcohol (PVA), polyvinyl ethers, polyvinyl esters, e.g., poly(vinyl acetate), polyvinyl halides, e.g., poly(vinyl chloride) (PVC), polyvinylpyrrolidone, polysiloxanes, polystyrene (PS), celluloses, including derivatized celluloses, e.g., alkyl celluloses, hydroxyalkyl celluloses, Examples of suitable polymeric materials include, but are not limited to, cellulose, cellulose ethers, cellulose esters, nitrocellulose, hydroxypropyl cellulose, and carboxymethyl cellulose, polymers of acrylic acid such as poly(methyl(meth)acrylate) (PMMA), poly(ethyl(meth)acrylate), poly(butyl(meth)acrylate), poly(isobutyl(meth)acrylate), poly(hexyl(meth)acrylate), poly(isodecyl(meth)acrylate), poly(lauryl(meth)acrylate), poly(phenyl(meth)acrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate) (together referred to herein as "polyacrylic acid"), polydioxanone and its copolymers, polyhydroxyalkanoates, polypropylene fumarate, polyoxymethylene, poloxamer, poly(butyric acid), trimethylene carbonate, polyphosphazenes, polysaccharides, peptides or proteins, and blends thereof.

[0148] In some embodiments, the one or more biocompatible polymers forming the nanoparticles, microparticles, or combinations thereof include FDA-approved biodegradable polymers, such as polyhydroxy acids (e.g., PLA, PLGA, and PGA), polyanhydrides, and polyhydroxyalkanoates, such as poly(3-butyrate) and poly(4-butyrate).

[0149] Materials other than polymers can be used to form nanoparticles, microparticles, or combinations thereof. Suitable materials include surfactants. The use of surfactants in nanoparticles, microparticles, or combinations thereof can improve surface properties, for example, by reducing particle-to-particle interactions, making the particle surface less adhesive. Both naturally occurring surfactants and synthetic surfactants can be incorporated into nanoparticles, microparticles, or combinations thereof. Exemplary surfactants include, but are not limited to, phosphoglycerides such as phosphatidylcholine (e.g., L-α-phosphatidylcholine dipalmitoyl), diphosphatidylglycerol, hexadecanol, fatty alcohols, polyoxyethylene-9-lauryl ether, fatty acids such as palmitic acid and oleic acid, sorbitan trioleate, glycocholate, surfactin, poloxamers, sorbitan fatty acid esters such as sorbitan trioleate, tyloxapol, and phospholipids.

[0150] Nanoparticles, microparticles, or a combination thereof may contain multiple layers. These layers may have similar or different release kinetic profiles for the active ingredient. For example, nanoparticles, microparticles, or a combination thereof may have a controlled release core surrounded by one or more additional layers. The one or more additional layers may preferably include an immediate release layer on the surface of the nanoparticles, microparticles, or a combination thereof. The immediate release layer may provide a bolus of active ingredient immediately after administration.

[0151] The composition and structure of the nanoparticles, microparticles, or combinations thereof can be selected so that the nanoparticles, microparticles, or combinations thereof are pH-sensitive or pH-responsive. In some embodiments, the nanoparticles, microparticles, or combinations thereof are formed from pH-sensitive or pH-responsive polymers, such as enteric polymers commercially available under the trademark EUDRAGIT®, as described above. Depending on the particle material, the nanoparticles, microparticles, or combinations thereof may undergo disintegration / degradation or structural changes at different locations in the gastrointestinal tract. In some embodiments, the particle material is selected so that the nanoparticles, microparticles, or combinations thereof can survive exposure to gastric acid after oral administration and release the active ingredient in the intestine.

[0152] D. Controlled Release

[0153] In some embodiments, the pharmaceutical formulation may be a controlled release formulation. Examples of controlled release formulations include extended release formulations, delayed release formulations, and pulsatile release formulations.

[0154] 1. Extended release

[0155] In some embodiments, extended release formulations are prepared as diffusion or osmotic systems, for example, as described in "Remington - The science and practice of pharmacy" (20th Ed., Lippincott Williams & Wilkins, 2000).

[0156] Diffusion systems are typically in the form of a matrix, generally prepared by combining active ingredients with a slowly dissolving carrier to form a tablet as needed.Suitable types of materials used in the preparation of the matrix include plastics, hydrophilic polymers, and fatty compounds.Suitable plastics include, but are not limited to, methyl acrylate-methyl methacrylate copolymer, polyvinyl chloride, and polyethylene.Suitable hydrophilic polymers include, but are not limited to, cellulose polymers such as methylethyl cellulose, hydroxyalkyl cellulose (e.g., hydroxypropyl cellulose and hydroxypropylmethyl cellulose), sodium carboxymethyl cellulose, CARBOPOL® 934, polyethylene oxide, and combinations thereof.Suitable fatty compounds include, but are not limited to, various waxes, such as carnauba wax and glyceryl tristearate, wax-type substances such as hydrogenated castor oil and hydrogenated vegetable oil, and combinations thereof.

[0157] In some embodiments, the plastic is a pharmaceutically acceptable acrylic polymer selected from acrylic acid and methacrylic acid copolymers, methyl methacrylate copolymers, ethoxyethyl methacrylate copolymers, cyanoethyl methacrylate copolymers, aminoalkyl methacrylate copolymers, poly(acrylic acid), poly(methacrylic acid), methacrylic acid alkylamine copolymers, poly(methyl methacrylate), poly(methacrylic acid), polymethacrylate, polyacrylamide, poly(methacrylic anhydride), and glycidyl methacrylate copolymers.

[0158] In some embodiments, the pharmaceutically acceptable acrylic polymer can be an ammonio methacrylate copolymer, which is well known in the art and is described as a fully polymerized copolymer of acrylic and methacrylic acid esters with a low content of quaternary ammonium groups.

[0159] In some embodiments, the pharmaceutically acceptable acrylic polymer is an acrylic lacquer, such as one commercially available under the trademark EUDRAGIT®. In some embodiments, the pharmaceutically acceptable acrylic polymer contains a mixture of two acrylic lacquers: EUDRAGIT® RL (e.g., EUDRAGIT® RL30D) and EUDRAGIT® RS (EUDRAGIT® RS30D). EUDRAGIT® RL30D and EUDRAGIT® RS30D are copolymers of acrylic and methacrylic acid esters with a low content of quaternary ammonium groups, with the molar ratio of ammonium groups to remaining neutral methacrylic acid esters being 1:20 for EUDRAGIT® RL30D and 1:40 for EUDRAGIT® RS30D. The code designations RL (high permeability) and RS (low permeability) refer to the permeability properties of these polymers. The EUDRAGIT® RL / RS mixture is insoluble in water and digestive fluids. However, multiparticulate systems formed containing the mixture are swellable and permeable to aqueous solutions and digestive fluids. The EUDRAGIT® RL / RS mixture can be prepared in any desired ratio to ultimately obtain a sustained-release formulation with a desired release profile. Suitable sustained-release multiparticulate systems can be obtained, for example, from 90% EUDRAGIT® RL + 10% EUDRAGIT® RS, to 50% EUDRAGIT® RL + 50% EUDRAGIT® RS, and 10% EUDRAGIT® RL + 90% EUDRAGIT® RS. In some embodiments, the pharmaceutically acceptable acrylic polymer can also be or include other acrylic resin lacquers, such as EUDRAGIT® S-100, EUDRAGIT® L-100, and mixtures thereof.

[0160] Matrices with different release mechanisms or profiles can be combined into a final dosage form containing a single unit or multiple units.Examples of multiple units include, but are not limited to, multi-layer tablets and capsules containing beads, granules, and / or fine granules of active ingredients.The immediate-release portion can be added to the extended-release system by applying an immediate-release layer on top of the extended-release core using a coating process or compression process, or in a multiple-unit system, such as a capsule containing both extended-release beads and immediate-release beads.

[0161] Extended-release tablets containing one or more hydrophilic polymers can be prepared by techniques commonly known in the art, such as direct compression, wet granulation, and dry granulation processes.

[0162] Extended-release tablets containing one or more fatty compounds can be prepared using methods known in the art, such as direct blending, coagulation, and aqueous dispersion methods. In the coagulation method, the active ingredient is mixed with the fatty compound, and either spray-coagulated or coagulated, sieved, and processed.

[0163] Alternatively, extended-release formulations can be prepared using osmotic systems or by applying semi-permeable coatings to solid dosage forms, in which case the desired release profile can be achieved by combining low-permeability and high-permeability coating materials in appropriate proportions.

[0164] 2. Delayed release

[0165] Delayed-release formulations can be prepared by coating a solid dosage form with a coating. In some embodiments, the coating is insoluble and impermeable in the acidic environment of the stomach and soluble or permeable in the less acidic environment of the intestine and / or lower gastrointestinal tract. In some embodiments, the solid dosage form is a tablet for incorporation into a capsule, a tablet for use as the inner core of a "coated core" dosage form, or a plurality of beads, granules, and / or fine granules containing the active ingredient for incorporation into either a tablet or a capsule.

[0166] Suitable coating materials include biodegradable polymers, gradually hydrolyzable polymers, gradually water-soluble polymers, and enzymatically degradable polymers, such as those described above. In some embodiments, the coating material is or contains an enteric polymer. A combination of different coating materials can also be used. Multi-layer coatings using different coating materials can also be applied.

[0167] Suitable weights for the coating or coating material can be readily determined by one skilled in the art by evaluating the individual release profiles of formulations prepared with different amounts of coating material.

[0168] The coating material may also contain one or more conventional additives, such as plasticizers (optionally about 10 wt% to 50 wt% based on the dry weight of the coating material), colorants, stabilizers, glidants, etc., such as those previously described.

[0169] 3. Pulsed release

[0170] Pulsed release formulations release multiple doses of active ingredient at spaced time intervals.Generally, when pulsated release formulations are administered, for example, orally, the initial dose is released substantially immediately, and for example, the first release "pulse" occurs within about 3 hours, 2 hours, or 1 hour after administration.This initial pulse can be followed by a first time interval (lag time) during which the active ingredient is released very little or not at all from the formulation, and then the second dose can be released.Similarly, a second lag time (almost no release interval) can be designed between the second release pulse and the third release pulse.The duration of the lag time varies depending on the formulation design, particularly depending on the length of the administration interval, for example, twice-daily administration profile, three times-daily administration profile, etc.

[0171] For pulsed release formulations that provide a twice-daily dosing profile, the formulations deliver two release pulses of the active ingredient. In some embodiments, the near-zero release interval between the first and second release pulses can have a duration of between 3 hours and 14 hours.

[0172] For pulsed release formulations that provide a three times daily dosing profile, the formulations deliver three release pulses of the active ingredient. In some embodiments, the negligible release interval between two adjacent pulses can have a duration of between 2 and 8 hours.

[0173] In some embodiments, the pulsatile release formulation contains multiple pharmaceutically acceptable carriers with different release kinetics.

[0174] In some embodiments, the pulsed release formulation contains a pharmaceutically acceptable carrier having multiple layers loaded with active ingredients. In some embodiments, the layers can have different release kinetics. In some embodiments, the layers can be separated by a delayed-release coating. For example, the pulsed release formulation can have a first layer for the first release pulse, with the active ingredient loaded on the surface, and a second layer, such as a core, for the second release pulse, with the active ingredient loaded on the second layer, and the second layer can be surrounded by a delayed-release coating, thereby creating a delay between the two release pulses.

[0175] In some embodiments, a pulsatile release profile is achieved using a formulation that is a closed, optionally sealed, capsule containing at least two "dosage units," where each dosage unit in the capsule provides a different release profile. In some embodiments, at least one of the dosage units is a delayed-release dosage unit. Control of the delayed-release dosage unit can be achieved by a controlled-release polymer coating on the dosage unit or by incorporating the active ingredient into a controlled-release polymer matrix. In some embodiments, each dosage unit can comprise a compressed or molded tablet, where each tablet in the capsule provides a different release profile.

[0176] E. Exemplary Formulations for Different Routes of Administration

[0177] Subjects suffering from the conditions, disorders, or diseases described herein can be treated by either targeted or systemic administration of pharmaceutical formulations containing the compounds or compositions described herein, via oral, inhalation, topical, transmucosal or submucosal, subcutaneous, parenteral, intramuscular, intravenous, or transdermal administration. In some embodiments, the pharmaceutical formulation is suitable for oral administration. In some embodiments, the pharmaceutical formulation is suitable for inhalation or intranasal administration. In some embodiments, the pharmaceutical formulation is suitable for transdermal or topical administration. In some embodiments, the pharmaceutical formulation is suitable for subcutaneous, intravenous, intraperitoneal, intramuscular, parenteral, or submucosal administration.

[0178] In some embodiments, the pharmaceutical preparation is an oral pharmaceutical preparation. In some embodiments, the active ingredient is incorporated with one or more of the aforementioned pharmaceutically acceptable excipients and can be used in the form of tablets, pills, caplets, or capsules. For example, the corresponding oral pharmaceutical preparation can contain one or more of the following pharmaceutically acceptable excipients, or those of similar nature: the aforementioned binders, disintegrants, lubricants, glidants, sweeteners (e.g., sucrose and saccharin), and flavoring agents (e.g., methyl salicylate and fruit flavors). In some embodiments, when the oral pharmaceutical preparation is in the form of a capsule, the preparation can contain a liquid carrier (e.g., fatty oil) in addition to the materials listed above. In some embodiments, when the oral pharmaceutical preparation is in the form of a capsule, each capsule can contain multiple beads, granules, and / or fine particles of the active ingredient. In some embodiments, the oral pharmaceutical formulation may contain one or more other materials that modify the physical form or one or more pharmaceutical properties of the dosage unit, for example, a coating of polysaccharides, shellac, or enteric polymers described in the previous section.

[0179] In some embodiments, oral pharmaceutical formulations may be in the form of an elixir, suspension, syrup, wafer, chewing gum, etc. A syrup may contain, in addition to the active ingredient, one or more sweetening agents (e.g., sucrose and saccharin), one or more flavoring agents, one or more preservatives, and / or one or more dyes or coloring agents.

[0180] In some embodiments, the pharmaceutical preparation is a parenteral pharmaceutical preparation. In some embodiments, the parenteral pharmaceutical preparation can be enclosed in an ampoule, a syringe, or a single- or multi-dose vial made of glass or plastic. In some embodiments, the parenteral pharmaceutical preparation is an intravenous pharmaceutical preparation. In some embodiments, the intravenous pharmaceutical preparation contains a pharmaceutically acceptable liquid carrier for the active ingredient. Suitable pharmaceutically acceptable liquid carriers include, but are not limited to, saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, NJ), phosphate buffered saline (PBS), and combinations thereof.

[0181] In some embodiments, the pharmaceutical preparation is a topical pharmaceutical preparation.Suitable forms of topical pharmaceutical preparations include lotions, suspensions, ointments, creams, gels, tinctures, sprays, powders, pastes, slow-release transdermal patches, and suppositories for application to the mucous membranes of the rectum, vagina, nose, or mouth.In some embodiments, thickeners, emollients (e.g., mineral oil, lanolin and its derivatives, and squalene), moisturizers (e.g., sorbitol), and / or stabilizers can be used to prepare topical pharmaceutical preparations.Examples of thickeners include petrolatum, beeswax, xanthan gum, and polyethylene.

[0182] In some embodiments, the pharmaceutical preparation is an intranasal pharmaceutical preparation.In some embodiments, the intranasal pharmaceutical preparation is in the form of an aqueous suspension, which can be placed in a pump spray bottle if necessary.Aqueous suspension can contain, in addition to water, one or more pharmaceutically acceptable excipients, such as suspending agents (e.g., microcrystalline cellulose, sodium carboxymethylcellulose, hydroxypropyl-methylcellulose), humectants (e.g., glycerol and propylene glycol), acid, base, and / or pH adjusting pH buffers (e.g., citric acid, sodium citrate, phosphoric acid, sodium phosphate, and combinations thereof), surfactants (e.g., Polysorbate 80), and preservatives (e.g., benzalkonium chloride, phenylethyl alcohol, and potassium sorbate).

[0183] In some embodiments, the pharmaceutical formulation is an inhalation pharmaceutical formulation. In some embodiments, the inhalation pharmaceutical formulation can be in the form of an aerosol suspension, a dry powder, or a liquid suspension. The inhalation pharmaceutical formulation can be prepared for delivery as a nasal spray or inhaler, for example, a metered dose inhaler (MDI). In some embodiments, the MDI can deliver aerosolized particles suspended in chlorofluorocarbon propellants, such as CFC-11 and CFC-12, or non-chlorofluorocarbon or alternative propellants, such as fluorocarbons (e.g., HFC-134A, HFC-227, etc.), with or without surfactants or suitable crosslinkers. Dry powder inhalers may also be used, and can be either breath-activated or pressure-driven for delivery.

[0184] In some embodiments, the active ingredients are prepared with pharmaceutically acceptable carriers that protect the active ingredients against rapid degradation or elimination from the subject's body after administration, such as controlled-release formulations described in the previous section.

[0185] V. How to use

[0186] Disclosed are methods for treating a condition, disorder, or disease in a subject in need thereof, comprising administering to the subject an effective amount of a compound, composition, or pharmaceutical formulation disclosed herein.

[0187] The compound, composition, or pharmaceutical formulation can be administered in various ways depending on whether local or systemic administration is desired. In some embodiments, the compound, composition, or pharmaceutical formulation is administered directly to a specific body location of a subject, for example, by topical administration and intranasal administration. In some embodiments, the compound, composition, or pharmaceutical formulation is administered systemically, for example, by enteral administration (e.g., oral administration) and parenteral administration (e.g., injection, infusion, and implantation). Exemplary administration routes include oral administration, intravenous administration, for example, intravenous injection or infusion, intranasal administration, and topical administration. In some embodiments, the compound, composition, or pharmaceutical formulation is administered orally. In some embodiments, the compound, composition, or pharmaceutical formulation is administered intravenously. In some embodiments, the compound, composition, or pharmaceutical formulation is administered intranasally.

[0188] In some embodiments, the subject is a human. In some embodiments, the subject is a human under the age of 18. In some embodiments, the subject is a non-human animal, such as a domestic pet, livestock and domestic animals, and zoo animals. In some embodiments, the non-human animal may be a non-human primate.

[0189] A. Indications

[0190] Normal synaptic transmission does not produce detectable acidification.Rather, excitatory synaptic transmission typically produces short-term alkalinization (Tong, et al., J Neurophysiol, 2006,95:3686-97; Makani and Chesler, J Neurosci, 2007,27:7438-7446).Therefore, under normal excitatory synaptic transmission, the compounds disclosed herein do not exhibit their pH sensitivity to any appreciable extent.In addition, the reduction of extracellular pH does not usually occur in the extrasynaptic NMDAR in normal brain.Therefore, the compounds disclosed herein are less effective in inhibiting GluN2B-containing NMDAR under normal conditions.

[0191] The pH sensitivity and high efficacy of the compound disclosed herein are suitable for the condition, disorder and disease that involves the acidification of the extracellular environment of GluN2B-containing NMDAR.In particular, the pH sensitivity of compound can be useful in a range of indications that can cause local acidification in the brain, such as stroke and subarachnoid hemorrhage.

[0192] The enhanced potency of compounds at GluN2B-containing NMDARs in an acidified extracellular environment can facilitate their neuroprotective effects after acute injury (e.g., ischemia). - and H + Ischemia, driven by both elevated CO2 and a shift to anaerobic metabolism with lactate production, typically decreases pH throughout the extracellular space. These mechanisms, which are potent drivers of infarct and penumbra acidification during ischemia, can affect both synaptic and nonsynaptic GluN2B-containing NMDARs.

[0193] The utility of the compounds of the present disclosure may also be applied to conditions, disorders, and diseases involving high frequency firing of nerve cells resulting in metabolic pH changes and local acidification, such as inflammatory pain.

[0194] Exemplary conditions, disorders, and diseases that can be treated by the disclosed compounds, compositions, and formulations include, but are not limited to, stroke, subarachnoid hemorrhage, cerebral ischemia, cerebral vasospasm, hypoxia, acute CNS injury, spinal cord injury, traumatic brain injury, coronary artery bypass graft, persistent or chronic cough, substance abuse disorders, opiate withdrawal, opiate tolerance, bipolar disorder, suicidal ideation, pain, fibromyalgia, depression, postpartum depression, resting tremor, dementia, epilepsy, seizure disorders, movement disorders, and neurodegenerative diseases.

[0195] In some embodiments, the condition, disorder or disease is selected from pain, depression, stroke, and subarachnoid hemorrhage.

[0196] In some embodiments, the condition, disorder or disease is stroke.In some embodiments, the compound, composition or pharmaceutical preparation is used to treat or prevent stroke-related damage.In some embodiments, the compound, composition or pharmaceutical preparation is administered under emergency care for stroke, stroke maintenance treatment, and / or stroke rehabilitation.

[0197] In some embodiments, the condition, disorder or disease is subarachnoid hemorrhage (SAH), for example, aneurysmal SAH. In some embodiments, the compound, composition or pharmaceutical preparation is used to treat or prevent SAH-related damage. In some embodiments, the compound, composition or pharmaceutical preparation is administered under emergency care for SAH, maintenance treatment of SAH, and / or rehabilitation of SAH.

[0198] SAH refers to an abnormal condition in which blood accumulates under the arachnoid membrane, the membrane that covers the brain. This area is called the subarachnoid space and normally contains cerebrospinal fluid. Blood accumulation in the subarachnoid space and the resulting vasospasm of blood vessels can lead to stroke, seizures, and other complications. SAH can occur naturally or be caused by head injury. Compounds, compositions, or pharmaceutical preparations can be used to treat subjects experiencing SAH. For example, compounds, compositions, or pharmaceutical preparations can be used to prevent or limit one or more of the toxic effects of SAH, including stroke and ischemia, which can result from SAH. Alternatively, compounds, compositions, or pharmaceutical preparations can be used to treat subjects with traumatic subarachnoid hemorrhage caused by head injury.

[0199] In certain embodiments, the compounds, compositions, or pharmaceutical preparations can be used to ameliorate neurological deficits resulting from SAH, e.g., aneurysmal SAH. In certain embodiments, the compounds, compositions, or pharmaceutical preparations are administered early in the treatment of the condition, e.g., approximately simultaneously with surgery to stop cranial bleeding. Delayed cerebral ischemia (DCI) occurs in approximately 30% of cases after aneurysmal SAH. In certain embodiments, the compounds, compositions, or pharmaceutical preparations are administered to prevent DCI associated with SAH. In certain embodiments, the compounds, compositions, or pharmaceutical preparations are administered during the period when the risk of DCI is highest, e.g., 3 to 14 days after the initial hemorrhage.

[0200] In some embodiments, the condition, disorder, or disease is pain. In some embodiments, the pain is chronic pain. In some embodiments, the pain is cancer pain. In some embodiments, the pain is neuropathic pain. Examples of neuropathic pain include diabetic peripheral neuropathy, postherpetic neuralgia, complex regional pain syndrome, peripheral neuropathy, rheumatoid arthritis, chemotherapy-induced neuropathic pain, cancer neuropathic pain, neuropathic lower back pain, HIV neuropathic pain, trigeminal neuralgia, and central post-stroke pain.

[0201] In some embodiments, neuropathic pain can be caused by trauma, ischemia, infection (e.g., HIV infection, shingles, and postherpetic neuralgia), metabolic and endocrinological disorders (e.g., diabetes mellitus, diabetic neuropathy, amyloidosis, and amyloid polyneuropathy (primary and familial)), vasculitic neuropathy, neuropathy associated with Guillain-Barré syndrome, neuropathy associated with Fabry disease, entrapment due to anatomical abnormalities, trigeminal neuralgia and other CNS neuralgias, malignant neuropathy, and rheumatoid arthritis. They may arise from peripheral or CNS pathological events, including, but not limited to, primary tumors, cryptic causes (e.g., idiopathic peripheral small fiber neuropathies), inflammatory conditions or autoimmune disorders (e.g., demyelinating inflammatory disorders, rheumatoid arthritis, systemic lupus erythematosus, and Sjogren's syndrome), compression of nerve fibers (e.g., radiculopathy and carpal tunnel syndrome), exposure to toxins or drugs, dietary or absorption abnormalities, gammopathy, and genetic abnormalities and amputations (including mastectomy).

[0202] In some embodiments, the condition, disorder, or disease is depression or postpartum depression, hi some embodiments, the depression is treatment-resistant depression.

[0203] In some embodiments, the condition, disorder or disease is a neurodegenerative disease. In some embodiments, the neurodegenerative disease is Huntington's disease, Alzheimer's disease, or Parkinson's disease. In some embodiments, the compound, composition, or pharmaceutical preparation is used to reduce one or more symptoms of a neurodegenerative disease. Exemplary symptoms include dementia (related to Alzheimer's disease) and dystonia and related movement disorders (related to Parkinson's disease). In some embodiments, the compound, composition, or pharmaceutical preparation is used to enhance cognition in subjects suffering from a neurodegenerative disease.

[0204] In some embodiments, the condition, disorder, or disease is epilepsy or a seizure disorder. In some embodiments, the subject's epilepsy or seizure disorder in need of treatment may include epilepsy that is not adequately controlled by existing pharmaceuticals (i.e., treatment-resistant epilepsy), infantile spasticity, and epilepsy or seizure disorders caused by rare diseases or genetic conditions (e.g., genetic mutations) that result in epilepsy, seizures, spasticity, abnormal hypersynchronous brain activity, and / or other conditions associated with enhanced synchrony of neural activity. In some embodiments, the subject may be a pediatric patient suffering from epilepsy or a seizure disorder. In some embodiments, the compound, composition, or pharmaceutical formulation is used to reduce the severity and / or intensity of the subject's epilepsy or seizure disorder. In some embodiments, the compound, composition, or pharmaceutical formulation is used to reduce the frequency of the subject's epilepsy or seizure disorder.

[0205] In some embodiments, the condition, disorder, or disease is dementia, hi some embodiments, the dementia is AIDS-induced dementia.

[0206] In some embodiments, the condition, disorder, or disease is hypoxia. In some embodiments, the compound, composition, or pharmaceutical preparation is used to treat or prevent hypoxia-related damage. In some embodiments, the compound, composition, or pharmaceutical preparation is administered in emergency care for hypoxic events, maintenance treatment of hypoxia, and / or rehabilitation of hypoxia. In some embodiments, the hypoxia is induced by respiratory failure, prolonged use of a ventilator, or both. In some embodiments, the respiratory failure, prolonged use of a ventilator, or both, are associated with COVID-19, including hospitalization caused by COVID-19.

[0207] In some embodiments, the condition, disorder or disease is cerebral ischemia.In some embodiments, the compound, composition or pharmaceutical preparation is used to treat or prevent cerebral ischemia-related damage.In some embodiments, the compound, composition or pharmaceutical preparation is administered under emergency care for cerebral ischemic events, maintenance treatment of cerebral ischemia, and / or rehabilitation of cerebral ischemia.In some embodiments, the cerebral ischemia is caused by traumatic brain injury, coronary artery bypass graft, carotid artery angioplasty, or neonatal ischemia after hypothermic circulatory arrest.

[0208] In some embodiments, the condition, disorder, or disease is cerebral vasospasm. In some embodiments, the cerebral vasospasm is caused or induced by SAH.

[0209] B. Dosage and Administration

[0210] In some embodiments, the compound, composition, or pharmaceutical formulation is administered for a period sufficient to alleviate one or more undesirable symptoms and / or one or more clinical signs associated with the condition, disorder, or disease being treated. In some embodiments, the compound, composition, or pharmaceutical formulation is administered less than three times per day. In some embodiments, the compound, composition, or pharmaceutical formulation is administered once or twice per day. In some embodiments, the compound, composition, or pharmaceutical formulation is administered once per day. In some embodiments, the compound, composition, or pharmaceutical formulation is administered in a single oral dosage once per day. In some embodiments, the compound, composition, or pharmaceutical formulation is administered in a single intravenous dosage once per day.

[0211] The dose of the compound per administration can be between 5 and 300 mg or as described above. In some embodiments, the dose of the compound per administration is between 25 and 200 mg. In some embodiments, the dose of the compound per administration is between 25 and 175 mg. In some embodiments, the dose of the compound per administration is between 25 and 150 mg. In some embodiments, the dose of the compound per administration is between 50 and 200 mg. In some embodiments, the dose of the compound per administration is between 75 and 200 mg. In some embodiments, the dose of the compound per administration is between 50 and 175 mg. In some embodiments, the dose of the compound per administration is between 75 and 150 mg.

[0212] In certain embodiments, the compound, composition or pharmaceutical preparation is administered at a loading dose of the compound per day for one or several days, and then at a reduced or normal dose of the compound per day for one or several days to complete the treatment course.For example, the compound, composition or pharmaceutical preparation is administered at a loading dose of the compound on the first day, and then at a reduced or normal dose per day for the remainder of the course.A suitable loading dose of the compound can be selected from the above-mentioned exemplary total daily doses.A suitable reduced or normal dose of the compound can also be selected from the above-mentioned exemplary total daily doses.In certain embodiments, the loading dose of the compound is about 150 mg, and the reduced or normal dose of the compound is 100 mg.For example, the compound, composition or pharmaceutical preparation is administered at a loading dose of 150 mg of the compound on the first day, and then at a reduced or normal dose of 100 mg of the compound per day for the remainder of the course. [Example]

[0213] The following examples describe studies to make and evaluate GluN2B-selective negative allosteric NMDAR modulators that have enhanced potency for GluN2B at pH 6.9 compared to pH 7.6.

[0214] Example 1 Synthesis and Characterization of Exemplary Compounds [ka]

[0215] A. Synthesis Procedure A suspension of (R)-6-(oxiran-2-ylmethoxy)-3,4-dihydroquinolin-2(1H)-one (100 g, 0.456 mol) and 1-(4-(trifluoromethyl)phenyl)piperazine (105 g, 0.456 mol) in ethanol (1 L) was stirred at 75° C. for 21 hours while monitoring by HPLC. The reaction became a clear solution within 15 minutes at 75° C. The reaction mixture was cooled to 50° C., and the precipitated solid was filtered and washed with ethanol (200 mL). The collected solid was dried under vacuum to give the crude product (175 g, 85.3%).

[0216] Crude NP10679 (260 g from multiple batches) was placed in a 5 L round-bottom flask and a premixed solution of methanol:acetone (1:1) was added with constant stirring. The suspension was heated to 50 °C with stirring until it became clear (approximately 30 min) and then filtered through a 2 μM filter. The clear solution was cooled to 30 °C over 15 min and added to water (13 L) with vigorous stirring over 10 min. The precipitated solid was stirred at 30 °C for 30 min, filtered, washed with water (7.8 L), and dried in a vacuum tray dryer at 70 °C for 48 h. This recrystallization yielded 255 g of a white solid (98% yield). The purity and chiral purity of the recrystallized product were determined to be >99% (by HPLC) and >98% (by chiral HPLC), respectively.

[0217] NP10309 was synthesized using an analogous method with (S)-6-(oxiran-2-ylmethoxy)-3,4-dihydroquinolin-2(1H)-one and 1-(4-(trifluoromethyl)phenyl)piperazine as starting materials.

[0218] Other compounds in Tables 1 and 2 were synthesized using methods similar to those described above, as well as those described in U.S. Patent No. 8,420,680 and Wang et al., Neurocrit Care, 2014, 20:119-131. In general, the chiral centers of the compounds were generated by ring-opening reactions of the corresponding epoxides.

[0219] For example, the synthesis of benzyl urea-containing compound 10075 was described in Wang et al., Neurocrit Care, 2014, 20:119-131. Other benzyl urea-containing compounds, including 10131, 10165, 10166, 10189, 10214, 10215, 10222, 10224, 10225, 10272, and 10294, were synthesized in the same manner.

[0220] The synthesis of phenol-containing compound 10045 was described in U.S. Patent No. 8,420,680. Other phenol-containing compounds, including NP10030, 10039, 10040, 10052, 10082, 10171, 10235, 10243, 10244, 10245, 10247, and 10249, were synthesized in the same manner.

[0221] The synthesis of 10146, a benzimidazolinone-containing compound, was described in U.S. Patent No. 8,420,680. Other fused ring (bicyclic) compounds, such as 10228, were synthesized in the same manner.

[0222] B. Chemical Characterization NP10679: 1 H NMR (400 MHz, DMSO-d6): δ 9.90 (brs, 1H), δ 7.50 (d, J=16Hz,1H), 7.05 (d, J = 16 Hz, 1H), 6.85 - 6.70 (m, 3H), 4.90 (brd, 1H), 4.00 - 3.80 (m, 3H), 3.30 - 3.20 (m, 4H), 2.90 - 2.75 (m, 2H), 2.70 - 2.30 (m, 8H). 13C NMR (75 MHz, CDCl3): δ 171.82, 154.63, 153.09, 131.21, 130.04, 126.36, 124.94, 122.87, 121.22, 120.79, 120.36, 119.92, 116.30, 114.52, 113.95, 113.24, 70.70, 65.80, 60.48, 53.02,47.98, 30.51, 25.58. 23 H 26 Calculated m / z for F3N3O3 500.47; found 500.30 [M+H].

[0223] Example 2 Measurement of GluN2B potency and pH dependence

[0224] A. Materials and Methods The GluN2B potency and pH dependence of NP10679, NP10309, and other compounds in Tables 1 and 2 were measured against the human GluN1-1a / GluN2B receptor (hereafter GluN1 / GluN2B) expressed in Xenopus oocytes, as measured by IC at pH 6.9 and 7.6, respectively. 50 The value was measured.

[0225] Two-electrode voltage-clamp recordings from Xenopus oocytes Stage V-VI unfertilized Xenopus oocytes were purchased from Ecocyte (Austin, Texas) and injected with 5 ng of GluN1 and 10 ng of GluN2B cRNA. Human GluN1 and GluN2B cDNAs encoding NCBI reference sequences NM_007327.3 and NM_000834.3, respectively, were linearized and cRNA was generated as previously described (Traynelis et al., J Neurosci 1998, 18(16):6163-75). After injection, oocytes were incubated in Barth's culture solution (88 mM NaCl, 1 mM KCl, 2.4 mM NaHCO3, 10 mM HEPES, 0.82 mM MgSO4, 0.33 mM Ca(NO3)2, 0.41 mM CaCl2, 10 U / mL PenStrep, and 0.1 mg / mL gentamicin, pH 7.4) at 18°C. HOLD Two-electrode voltage-clamp (TEVC) recordings were taken at 22–23°C 2–7 days after injection using a voltage (V) of −40 mV. Briefly, oocytes were perfused in recording solution (90 mM NaCl, 1 mM KCl, 10 mM HEPES, 0.01 mM EDTA, and 0.5 mM BaCl) adjusted to either pH 7.6 or 6.9 by the addition of NaOH or HCl, respectively (pH 6.9 solution was prepared by adding HCl to the pH 7.6 solution, resulting in equal concentrations of NaOH in both solutions). + Compound concentration-response curves were obtained by applying increasing concentrations of each individual compound until steady-state conditions were obtained in the presence of saturating agonist concentrations (i.e., 100 μM glutamate and 30 μM glycine). Typically, oocyte recordings were taken from 4–10 oocytes per experiment (i.e., oocyte injection cycle) from two or more experiments. The concentration-response relationship for each oocyte was fitted by equation (1): Percent response = (100 - min) / (1 + ([concentration] / IC 50 ) nH )+min (1) where min is the residual response at saturating concentrations of each individual compound (constrained to be ≥ 0), and IC50 is the concentration of compound that causes half-maximal inhibition and nH is the Hill slope.

[0226] NP10679 was also tested for activity at GluN2A (NM_000833), GluN2C (NM_000835), and GluN2D (NM_000836) NMDARs in a similar manner to GluN2B, except that NP10679 was tested at a single concentration of 3 μM.

[0227] B. Results The IC of compounds against GluN2B measured at pH 6.9 and 7.6 50 The values ​​are shown in Tables 1 and 2.

[0228] Table 1 shows the IC values ​​for GluN2B for nine pairs of enantiomers. 50 Among these compounds, the R enantiomers exhibited a much lower pH boost compared to their corresponding S enantiomers. Here, the pH boost of a particular compound is its IC determined at pH 6.9. 50 The IC determined at pH 7.6 for 50 It is defined as the ratio of the values

[0229] In Table 1, some R enantiomers, e.g., 10233, 10249, and 10228, exhibit lower potency (i.e., higher IC) against GluN2B compared to their corresponding S enantiomers. 50 ), while the other R enantiomers showed comparable or even higher potency against GluN2B compared to their corresponding S enantiomers. [Table 1-1] [Table 1-2] [Table 1-3]

[0230] Table 2 shows the IC values ​​for GluN2B for six pairs of enantiomers. 50 Among these compounds, the structure-activity relationships are quite different from those obtained from Table 1. In particular, the R enantiomers exhibited comparable or even higher pH boosts compared to their corresponding S enantiomers. Furthermore, all R enantiomers exhibited comparable or higher potency against GluN2B than their corresponding S enantiomers.

[0231] For example, NP10679 has an IC of 23 nM at pH 6.9. 50 value and an IC of 142 nM at pH 7.6 50 values, corresponding to a pH boost of 6.2. In comparison, its S enantiomer, NP10309, had an IC of 111 nM at pH 6.9. 50 value and an IC of 717 nM at pH 7.6 50 values, corresponding to a pH boost of 6.5. [Table 2-1] [Table 2-2]

[0232] Furthermore, the activity of NP10679 against GluN2A, GluN2C, and GluN2D was measured at pH 6.9. NP10679 is more selective for the GluN2B subunit than for GluN2A, GluN2C, and GluN2D. There was no notable off-target inhibition against GluN2A, GluN2C, and GluN2D at 3 μM (Table 3). [Table 3]

[0233] Example 3 In vitro drug profiling

[0234] A. Materials and Methods Liver microsomal stability, cytochrome P450 inhibition, and plasma protein binding

[0235] Metabolic stability was assessed using human and mouse liver microsomes (Xenotech, USA). The final assay composition contained 1 μM test compound or reference standard (imipramine and diclofenac sodium) prepared from DMSO or acetonitrile stocks to final concentrations of 0.2% and 0.8%, respectively. Test compounds were added to 0.5 mg / mL microsomal protein in 100 mM potassium phosphate buffer alone, pH 7.4, without cofactors (5.0 mM glucose-6-phosphate, 0.06 U glucose-6-phosphate dehydrogenase, 2.0 mM MgCl, 1.0 mM NADP). + / NADPH). Test compounds and standards were incubated with human and mouse liver microsomes at 37°C, and aliquots of the reaction mixture (100 μL) were removed at 0, 5, 15, 30, 60, and 120 minutes. The reaction in the aliquots was stopped by adding 2.5 mL of tert-butyl methyl ether, and the samples were shaken for 15 minutes. The samples were then spun at 4000 rpm at 10°C for 15 minutes, and the organic phase was evaporated to dryness and then reconstituted with solvent for LC-MS / MS analysis. The percentage of test compound remaining after the specified incubation period was calculated based on the peak area of ​​the test compound at time 0 minutes.

[0236] Inhibition of CYP2D6 and CYP3A4 was achieved using recombinant human isoforms and the Vivid CYP Blue Screening Kit (Invitrogen, USA) by incubating two-fold serial dilutions of test compounds (nine samples) with kit reagents and reaction buffer in a 96-well plate according to the manufacturer's protocol. The plate was then incubated at room temperature for 30 minutes, after which fluorescence was measured using a plate reader. For these studies, the reference standards ketoconazole (CYP3A4) and quinidine (CYP2D6) were used as controls.

[0237] Plasma protein binding was performed using a rapid equilibrium dialysis (RED) device containing a dialysis membrane with a molecular weight cutoff of 8,000 daltons according to the manufacturer's instructions (ThermoFisher, USA). Plasma sample (pH 7.4) and test compound solution (1 μM or 5 μM) or reference standard (warfarin and propranolol, 10 μM) were combined (final DMSO concentration 0.1%). 300 μL of this spiked plasma sample was added to the sample chamber, and 500 μL of blank PBS buffer (pH 7.4) was added to the buffer chamber. The RED device was sealed with adhesive film and then incubated at 37°C for 4 hours with shaking at 300 rpm. After incubation, aliquots (50 μL) were removed from each well (spiked plasma and buffer side), diluted with an equal volume of the corresponding matrix from the opposite side (blank buffer or blank plasma) to negate matrix effects, and then extracted for analysis by LC-MS / MS. The amount of free material was calculated using the following formula: Free % = (LC-MS / MS peak area of ​​test compound on buffer side / LC-MS / MS peak area of ​​test compound on plasma side) × 100% It was decided by.

[0238] Off-target screening The in vitro effects of NP10679 on hERG (human delayed rectifier potassium ion channel gene) potassium channel current (IKr, a surrogate for rapidly activating delayed rectifier cardiac potassium current) were evaluated at room temperature in HEK mammalian cells stably expressing hERG using a QPatch HT® (Sophion Bioscience A / S, Denmark) and an automated parallel patch clamp system (ChanTest, Cleveland, OH). NP10679 was diluted to 0.1 μM, 0.3 μM, 1 μM, and 3 μM in an HB-PS solution composed of (in mM): NaCl 137; KCl 4.0; CaCl 1.8; MgCl 1; HEPES 10; and glucose 10 (pH adjusted to 7.4). Each test concentration was tested in duplicate or more cells (n≧2). The duration of exposure to each test article concentration was 3 minutes. A positive control (0.5 μM E-4301) was used to confirm the sensitivity of the cells to the hERG inhibitor.

[0239] Off-target radioligand binding displacement studies of NP10679 were conducted at the National Institutes of Mental Health Psychoactive Drug Screening Program (NIMH PDSP) at the University of North Carolina at Chapel Hill. Briefly, compounds were submitted to the NIMH PDSP and screened at a single concentration (10 μM) of test article under equilibrium conditions for their ability to displace specific radioligand binding to targets expressed in mammalian cell membranes in vitro. Each receptor target was assayed in quadruplicate, and the % inhibition of radioligand binding at each target was determined at pH 7.4. If the % inhibition was >50%, a full competitive displacement binding study was performed to determine the IC 50 value and this IC 50 From the value, the Cheng-Prusoff equation (K i =IC 50 / [1+(L / K d )]) (L is the radioligand concentration used in the competitive binding assay, and K dis the radioligand equilibrium binding affinity determined in the saturation binding assay described above) using K i value was determined.

[0240] The following targets (radioligins in parentheses) were tested: 5-HT1A ([ 3 H]8-OH-DPAT), 5-HT1B([ 3 H]5-carboxyamidotryptamine), 5-HT1D ([ 3 H]5-carboxyamidotryptamine), 5-HT1E ([ 3 H]5HT), 5-HT2A ([ 3 H]ketanserin), 5-HT2B ([ 3 H]LSD), 5-HT2C ([ 3 H] mesulergine), 5-HT3 ([ 3 H]LY278584), 5-HT5A([ 3 H]LSD), 5-HT6([ 3 H]LSD), 5-HT7([ 3 H]LSD), Alpha 1A ([ 3 H]prazosin), alpha 1B ([ 3 H]prazosin), alpha 1D ([ 3 H]prazosin), alpha 2A ([ 3 H]-rauwolscine), alpha 2B ([ 3 H]-rauwolscine), alpha 2C ([ 3 H]-rauwolscine), beta 1([ 125 I] pindolol), beta 2 ([ 3 H]CGP12177), beta 3 ([ 3 H]CGP12177), BZP rat brain tissue ([ 3 H]flunitrazepam), D1([ 3 H]SCH23390), D2([ 3 H]N-methylspiperone), D3([ 3 H]N-methylspiperone), D4([ 3 H]N-methylspiperone), D5 ([ 3 H]SCH23390), DAT([ 3 H]WIN35428), DOR([ 3H]DADLE), GABAA([ 3 H]muscimol), H1([ 3 H]pyrilamine), H2([ 3 H] thiotidine), H3([ 3 H]alpha-methylhistamine), H4([ 3 H]histamine), KOR([ 3 H]U69593), M1([ 3 H]QNB), M2([ 3 H]QNB), M3([ 3 H]QNB), M4([ 3 H]QNB), M5([ 3 H]QNB), MOR([ 3 H]DAMGO), NET([ 3 H]nisoxetine), PBR([ 3 H]PK11195), SERT([ 3 H]citalopram), Sigma 1 ([ 3 H]pentazocine(+)), and Sigma 2 ([ 3 H]DTG).

[0241] Some receptor targets were also tested in functional studies to establish whether NP10679 acts as an agonist or antagonist. These receptor targets include 5-HT 2A To assess agonism, human 5-HT receptors were included and functional studies were performed at pH 7.4 (Porter, et al., Br J Pharmacol, 1999, 128:13-20; CEREP, France). 2AHEK293 cells transfected with NP10679 were incubated with increasing concentrations of NP10679 (duplicate wells / concentration) at 37°C for 30 minutes. Receptor activation was determined by changes in IP1 levels detected by the HTRF® method. Separate wells stimulated with 10 μM serotonin served as a positive control. To determine antagonism by NP10679, cells were incubated with increasing concentrations of the compound (duplicate wells per concentration) at 37°C for 30 minutes. Cells were stimulated with 100 nM serotonin. Receptor activation was determined by changes in IP1 levels detected by the HTRF® method. A control inhibitor, ketanserin, was run separately to confirm the accuracy and reliability of the assay data.

[0242] Human α-glucan was incubated with increasing concentrations of NP10679 (duplicate wells per concentration) at pH 7.4 at room temperature. 1A Similar studies were performed in CHO cells transfected with α-adrenergic receptors to assess agonism and antagonism (Vicentic, et al., J Pharmacol Exp Ther, 2002, 302:58-65). Receptor activation was measured by measuring intracellular changes [Ca] using a fura-2 fluorometric detection method (CEREP, France). 2+ As a positive control, separate wells were stimulated with 30 nM epinephrine. To assess antagonism, cells were incubated with increasing concentrations of NP10679 (duplicate wells / concentration) at room temperature, and then cells were stimulated with 3 nM epinephrine. Receptor activation was measured by fura-2 fluorimetric detection (CEREP, France) to measure intracellular changes [Ca]. 2+ ] was decided.

[0243] To assess agonism and antagonism at the human H1-histamine receptor, HEK293 cells transfected with H1 receptors were incubated with increasing concentrations of NP10679 (duplicate wells per concentration) at room temperature at pH 7.4 (Miller, et al., J Biomol Screen, 1999, 4(5):249-258). Receptor activation was measured by measuring intracellular changes [Ca] using fura-2 fluorimetric detection. 2+ ]. Separate wells were stimulated with 10 μM histamine as a positive control. To assess antagonism by NP10679, cells were incubated with increasing concentrations of the compound (duplicate wells per concentration) at room temperature, and then the cells were stimulated with 300 nM histamine. Receptor activation was measured by fura-2 fluorimetric detection of intracellular changes [Ca 2+ The control inhibitor, pyrilamine, was run separately to confirm the accuracy and reliability of the assay data (CEREP, France).

[0244] B. Results Metabolic stability was performed using human and mouse liver microsomes with 1 μM NP10679 prepared from a DMSO stock (final DMSO 0.2%). Compounds and standards were incubated with human and mouse liver microsomes with or without cofactors, and samples were extracted and analyzed using LC-MS / MS as described above. NP10679 showed excellent stability in both human and mouse liver microsomes; after 1 hour of incubation at 37°C in the presence of cofactors, 72% of NP10679 remained in the incubation with human microsomes and 54% remained in the incubation with mouse liver microsomes.

[0245] Furthermore, 1 μM NP10679 did not inhibit the human recombinant cytochrome 450 isoforms CYP3A4 or CYP2D6.

[0246] Furthermore, NP10679 bound to human, mouse, and dog plasma proteins at 97.7% (n=2), 98.2% (n=2), and 98.2% (n=1), respectively.

[0247] NP10679 was also tested at 10 μM for binding to 41 neurotransmitter receptors, enzymes, and channels by radioligand displacement in competitive receptor binding assays. Targets for which 10 μM NP10679 displaced more than 50% of the radioligand were followed up in full dose-effect displacement studies, thereby demonstrating that five of these targets, namely 5-HT 2A Serotonin receptor (0.638 μM), α 1A (0.603 μM) and α 1D Submicromolar K for adrenergic receptors (0.495 μM), H1 histamine receptors (0.040 μM), and the serotonin transporter SERT (0.135 μM) i The values ​​were determined. See Table 4. In addition, three receptors (5-HT 2A , α 1A The compounds (adrenaline, and H1 histamine) were tested for functional agonism and antagonism, and in all cases, the compounds behaved as antagonists (Table 4). Inhibition of the human delayed rectifier cardiac potassium current channel (hERG channel) was measured by patch clamp electrophysiology across four concentrations of NP10679 in mammalian HEK cells transfected with hERG potassium channel cDNA, thereby yielding an IC50 of 0.617 μM for inhibition. 50 was revealed (Table 4). [Table 4]

[0248] Example 4 In vivo efficacy and pharmacokinetic studies

[0249] A. Materials and Methods Formulation and drug administration

[0250] For MCAO, locomotor activity, and rotarod studies, NP10679, MK-801, and ifenprodil were formulated in 2% or 10% N',N'-dimethylacetamide, 10% propylene glycol, and 30% 2-hydroxypropyl-beta-cyclodextrin in water at a dose volume of 10 mL / kg and administered intraperitoneally (IP). For pharmacokinetic studies, formulations used 2% or 10% N',N'-dimethylacetamide, 10% propylene glycol, and 30% 2-hydroxypropyl-beta-cyclodextrin in water at a dose volume of 10 mL / kg (all routes of administration).

[0251] In vivo model of transient focal ischemia All animal protocols were approved by Georgia State University's IACUC, an AAALAC-accredited program, and performed under the supervision of a board-certified veterinarian. Mice were group-housed, provided with nesting materials and shelter, and given free access to food pellets and water under a 12-hour light / dark cycle. Mice were placed in individual rooms and allowed to rest for at least 30 minutes before the start of surgery.

[0252] Mice (C57B16, >90 days old, Jackson Labs) were subjected to transient (60 min) middle cerebral artery occlusion (MCAO), and infarct volume was measured 24 h after reperfusion as previously described (Yuan, et al., Neuron, 2015, 85(6):1305-1318). Male mice were used for this experiment to reduce potential confounding by progesterone fluctuations due to the estrous cycle, which may have neuroprotective effects. Briefly, transient ischemia was induced in anesthetized (2% isoflurane / 98% O2) mice by inserting an intraluminal suture into the MCA for 60 min (Junge, et al., Proc Natl Acad Sci USA, 2003, 100: 13019-13024). The body temperature of each mouse was monitored by rectal thermometer and maintained at 37 °C using a homeothermic blanket. Changes in regional cerebral blood flow were monitored with a laser Doppler flowmeter probe (Perimed) adhesively fixed to the skull 4–6 mm lateral and 2 mm posterior to bregma. A non-absorbable 11 mm 5-0 Dermalon or Look (SP185) black nylon suture with a flame-tipped rounded tip was introduced into the left internal carotid artery via the external carotid stump until the suture was inserted 10.5–11 mm. Only mice whose blood flow decreased to less than 20% within 60 min and recovered to more than 90% after suture removal were allowed to proceed to completion of the study. After the occlusion period, mice were returned to their cages on a heated blanket (37°C) for several hours and monitored for righting reflex and ability to ambulate when gently handled. At 24 hours after occlusion, mice were euthanized with an overdose of isoflurane, and brains were rapidly removed, cut into 2 mm slices, and incubated in 2% 2,3,5-triphenyltetrazolium chloride (TTC) in phosphate-buffered saline (pH 7.4) at 37°C for 20 minutes, then placed at 4°C for imaging. The infarct area was then measured using NIH IMAGE software (Scion Corporation, Beta 4.0.2 release). The lesion area of ​​each slice was determined by a digital threshold reduction of TTC staining to a level 20% lower than that observed in the contralateral cortex.The infarct area was then manually outlined with a cursor to determine the cubic volume of the infarct for each slice, and all four slices from each animal were then summed to obtain the total infarct volume. The ratio of the contralateral to ipsilateral hemispheric volume was multiplied by the corresponding infarct slice volume to correct for edema. Drugs were administered by IP injection 5 minutes before the start of surgery (approximately 15 minutes before vascular occlusion). All drug doses were randomized, and investigators were blinded throughout the study, from the surgical procedure through analysis of stained slices to measurement of infarct volume.

[0253] statistics Based on historical variability and an expected effect size of 45-50%, we estimated that n = 12 per group (4 groups per study) was adequate to detect significant effects (α = 0.05) with sufficient power (β = 0.90) (G*Power 3.1). Infarct volumes after administration of drug doses were compared with vehicle controls (p < 0.05) by one-way ANOVA and Dunnett's test.

[0254] Pharmacokinetic studies Pharmacokinetic studies with NP10679 were outsourced to Anthem Biosciences (Bangalore, India) and were performed in accordance with CPCSEA guidelines after obtaining approval from the Institutional Animal Ethics Committee (IAEC).

[0255] NP10679 properties were evaluated in male BALB / c mice (8-10 weeks old, 20-30 g). Briefly, mice were administered a 2 mg / kg or 5 mg / kg dose (n=3 each) via IP injection (dose volume 10 mL / kg). Blood samples were collected on ice into tubes containing sodium heparin at 0.08, 0.25, 0.5, 1, 2, 4, 8, and 24 hours post-dose. 100 μL of plasma was combined with 50 μL of internal standard (haloperidol, 10 μg / mL). The tubes were then spun at 4000 g for 10 minutes (4°C). The plasma was transferred to a clean tube and stored at -80°C until analysis. The analyte NP10679 was quantified by API 3200 Q-trap LC-MS / MS and compared to standards, and data were analyzed by WinNonlin 6.3 (Pharsight).

[0256] In a separate study, BALB / c mice were administered either an oral dose (10 mg / kg) or an intravenous dose (3 mg / kg) of NP10679 (injection volume 10 mL / kg). Blood samples were collected on ice in sodium heparin tubes at 0.08, 0.25, 0.5, 1, 2, 4, 8, and 24 hours post-dose. Samples were prepared and analyzed as described above, except that the internal standard was fluconazole (10 μg / mL).

[0257] NP10679 was also measured in the brain compartment relative to blood at 0.25 and 1 hour after an IV dose of 3 mg / kg in two separate studies using blood samples collected and prepared as described above. Here, brain samples were first washed with deionized water to remove blood, the weight recorded, then transferred to 1 mL of fresh water, homogenized, and stored at -80°C until analysis. The ratio of compound in brain (g) relative to blood (mL) was then calculated.

[0258] B. Results Previous development of nonselective NMDAR inhibitors, which blocked all NMDARs regardless of subunit composition, resulted in both off-target and on-target adverse effects that complicated or even halted clinical development. The most notable side effects reported included motor dysfunction, cognitive impairment, and psychotomimetic effects such as hallucinations and disorganized thought (Lees, et al., Lancet, 2000, 355:1949-1954; Sacco, et al., JAMA, 2001, 285:1719-1728; Diener, et al., J Neurol, 2002, 249:561-568; Rowland, Aviat Space Environ Med, 2005, 76:C52-C58; Blagrove, et al., Psychopharmacol, 2009, 203:109-120). GluN2B-selective NMDAR negative allosteric modulators appear to be better tolerated than competitive antagonists or channel blockers, but may still exhibit side effects (Chaperon, et al., Behav Pharmacol, 2003, 14:477-487; DeVry and Jentzsch, Behav Pharmacol, 2003, 14:229-235; Yurkewicz, et al., J Neurotrauma, 2005, 22:1428-1443; Nicholson, et al., Behav Pharmacol, 2007, 18:731-743; Preskorn, et al., J Clin Psychopharmacol, 2008, 28:631-637; Nutt, et al., Mov Disord, 2008, 23: 1860-1866).

[0259] In the MCAO experiments, transient ischemia was induced by occlusion of the middle cerebral artery after administration of NP10679. Vehicle-treated mice showed substantial neuronal death, and after 60 min of transient ischemia, the infarct volume was 101 ± 8.7 mm. 3In comparison, NP10679 reduced infarct volume in a dose-dependent manner, with an ED 50 At an IP dose of 1 mg / kg, the maximum infarct volume reduction was 52% (Figure 1). 3 ) and 10 mg / kg (49 ± 3.0 mm 3 ) doses significantly reduced infarct volume compared to vehicle controls (Figure 1).

[0260] In a pharmacokinetic study, mice were administered a solution orally (10 mg / kg) or by IV injection (3 mg / kg) to determine both oral bioavailability and plasma pharmacokinetics for NP10679 (Figure 2A, Table 5). The plasma terminal half-life for the oral route was 7.06 hours and for IV administration was 8.56 hours, with a high volume of distribution of 1.59 L / kg and clearance of 2.44 mL / min / kg, resulting in high oral bioavailability (75.7%). See Table 5.

[0261] In a separate study, mice were IP-administered 2 mg / kg and 5 mg / kg of NP10679 according to the same dose and route of administration used in the MCAO neuroprotection study to provide drug pharmacokinetic information in mice. Here, NP10679 demonstrated dose-dependence, with peak plasma levels 30 min after administration of 581 ng / mL and 1431 ng / mL, respectively, and a plasma half-life of 7.5–9.9 h (Table 5). Thus, a single IP administration of NP10679 provided sufficient exposure to promote neuroprotection for as long as 24 h after the ischemic period. Figure 2B shows the calculated free plasma levels (unbound drug) at both the 2 mg / kg and 5 mg / kg IP doses, indicating that the free drug levels after the 5 mg / kg dose corresponded to the IC value for GluN2B at pH 6.9. 50 The free plasma levels of NP10679 were calculated based on the free drug fraction determined by the plasma binding studies described above.

[0262] Furthermore, pharmacokinetic studies in the brain compartment demonstrated that NP10679 exhibited high brain penetration, with levels ranging from 1.3 to 2.6-fold higher in the brain compartment compared to plasma levels in mice 1 h after IV administration (Table 5). Based on these brain:plasma ratios, it is estimated that the free drug concentrations in the brain after the 5 mg / kg IP dose used in the MCAO study could reach 60-134 nM, 51-103 nM, 33-66 nM, and 28-56 nM at 1, 2, 4, and 8 h after administration, respectively. Considering that the potency of NP10679 for GluN2B at pH 6.9 is 23 nM, the occupancy of GluN2B receptors at pH 6.9 in the brain compartment is sufficiently high to drive significant GluN2B inhibition. [Table 5]

[0263] Example 5 Locomotor activity and rotarod performance

[0264] A. Materials and Methods Measurement of locomotor activity and rotarod performance

[0265] Locomotor activity and rotarod studies were approved by the IACUC at Georgia State University, an AAALAC-accredited facility, under the supervision of a board-certified veterinarian. Mice were group-housed, provided with nesting material and shelter, and given free access to food pellets and water under a 12-hour light / dark cycle.

[0266] For locomotor activity measurements, mice (C57Bl6, >90 days old, Jackson Labs) were placed in a closed (lighted) activity monitoring box for 1 hour prior to drug testing for acclimation. After 1 hour, the animals were removed, injected with drug (IP), and then returned to the activity monitoring box for 2 hours of total locomotor activity monitoring. The total number of light breaks (horizontal) in the cage was determined by computer, and the results were averaged for each drug. Results were analyzed by ANOVA and Dunnett's post-hoc test to compare horizontal activity of drug-treated groups to vehicle controls. Given that only male mice were used in the MCAO transient ischemia study, male animals were used in these behavioral studies.

[0267] For the rotarod experiment, male C57BL / 6 mice (>90 days old) were tested using a Rotamax 4 / 8 rotarod (Columbus Instruments, Columbus, Ohio). Prior to training and testing, mice were placed in the testing room and allowed to acclimate for 2 hours before any further handling. Mice were placed on a 3.8 cm diameter and 8 cm wide rotating rod (5 rpm) elevated 30 cm above the chamber floor. After 10 seconds of rotation at a fixed speed, rotation was slowly accelerated from 5 rpm to 35 rpm over 5 minutes. The duration that mice could remain on the rotarod without hanging or falling during a 360° rotation was recorded. Mice were trained four times daily for two days, with a 25-minute interval between tests. On the third day, mice were randomly assigned to treatment groups and tested 25 minutes after administration of the test drug or vehicle (IP administration) (four trials, with a 25-minute interval between tests). The individual conducting the experiment was blinded to the identity of each treatment group. Results were analyzed by ANOVA and Dunnett's post-hoc test to compare the duration on the rotorod of drug-treated groups with vehicle controls.

[0268] B. Results A study was conducted to assess whether NP10679 disrupts motor coordination or function. Mice were administered NP10679 and then tested in a rotarod challenge study. Here, mice were trained for two consecutive days, with four trials per day (with a 25-minute interval between trials), for their ability to remain on a rotating, accelerating bar. Mice demonstrated improved performance from day 1 to day 2 across trials conducted on a single day, as shown in Figure 3. On day 3, mice were randomly assigned to treatment groups, administered vehicle or drug, and then tested four times, starting 25 minutes after administration, and the mean latency to fall was established for each trial (Figure 3). Across all four trials, NP10679 did not significantly impair function when administered at 2 mg / kg or 5 mg / kg. The 10 mg / kg NP10679 dose group demonstrated a reduced latency to fall (87 ± 13 seconds) on the fourth trial compared to vehicle controls (168 ± 14 seconds). However, no statistically significant changes from vehicle controls were observed in this treatment group on the first, second, or third trials. In contrast, a 30 mg / kg dose of ifenprodil significantly reduced the latency to fall scores in all four trials tested (Figure 3). The higher dose was chosen because ifenprodil is less potent than NP10679 at GluN2B (Kew, et al., J Physiol, 1996, 497:761-772; Mott, et al., Nat Neurosci, 1998, 1(8):659-67) and higher concentrations are required to produce neuroprotection in vitro (Chenard, et al., J Med Chem, 1991, 34(10):3085-90).

[0269] The ability of a single dose of NP10679 to alter locomotor activity in mice was assessed in a closed, lighted chamber (Figure 4). After a 1-hour habituation period, mice were administered a 20 mg / kg dose of NP10679 or 0.3 mg / kg of MK-801, returned to the closed, lighted chamber, and horizontal activity was measured for 2 hours. This dose of NP10679 did not result in any statistically significant decrease in horizontal activity in mice compared to vehicle control (n = 6 each). In contrast, administration of 0.3 mg / kg of MK-801 resulted in a significant increase (p < 0.01) in horizontal activity (n = 4).

[0270] In summary, NP10679 exhibits enhanced inhibition of GluN2B at extracellular acidic pH values ​​(pH 6.9) relative to pH 7.6. In particular, NP10679 exhibits much higher potency against GluN2B compared to its S enantiomer (NP10309), while maintaining a pH boost.

[0271] Because glutamate-containing vesicles are acidic within their lumen, these properties make NP10679 a more effective inhibitor of NMDARs at synapses that respond to high-frequency action potentials compared with its S enantiomer. In addition, acidification of the border zone around ischemic tissue can also enhance the effects of NP10679 for improved neuroprotection.

[0272] When tested in cognitive tasks and learning paradigms after acute administration in non-human primates, two GluN2B inhibitors, namely traxoprodil and BMT-108908, produced dose-dependent cognitive impairment (Weed, et al., Neuropsychopharm, 2016, 46:568-577). Traxoprodil does not have significant pH sensitivity between pH 6.8 and pH 7.5 for receptor inhibition (Mott, et al., Nat Neurosci, 1998, 1(8):659-67). The high potency and significant pH-boosting effect of NP10679 on GluN2B offer advantages over existing GluN2B-targeted drugs and drug candidates in distinguishing side effects from desired on-target activity.

[0273] Furthermore, NP10679 has high oral bioavailability along with excellent brain penetration, making it suitable for both intravenous and oral administration for therapeutic use in humans.

[0274] Example 6 Human clinical studies

[0275] A. Materials and Methods Drug Substances and Products Synthesis of GMP-quality pharmacologically active ingredient (API) of NP10679 for use in the drug product was outsourced to DavosPharma (Saddle River, NJ). Drug product manufacturing was performed by the University of Iowa, Pharmaceuticals (UI-P) according to established procedures for lyophilized product fabrication. To formulate the drug product, the API was solubilized to a concentration of 5 mg / mL in a vehicle of 25% hydroxypropyl-beta-cyclodextrin (HPBCD) in 50 mM monobasic potassium phosphate buffer (pH 6.0). This solution was then filtered, sterilized, and lyophilized into sterile vials each containing 50 mg of API. The lyophilized API was formulated at the clinical site into a drug product for IV infusion by adding the appropriate amount of 2.5% HPBCD in 0.9% saline.

[0276] method The protocols for both the single ascending dose (SAD) and multiple ascending dose (MAD) studies were reviewed and approved by the U.S. Food and Drug Administration under an investigational new drug application. These protocols and subject informed consent packages were also reviewed and approved by the study's institutional review board (IRB), IntegReview IRB, Austin, TX. The clinical research organization (CRO) for both studies was Pharmaron CPC, Baltimore, Maryland. All subjects were informed of the nature and purpose of the studies, and written informed consent was obtained from subjects before any study-related procedures were performed. The studies were conducted in accordance with the principles outlined in the Declaration of Helsinki and the International Conference on Harmonization Tripartite guidelines, which are based on Good Clinical Practice.

[0277] Inclusion and Exclusion Criteria Healthy male and female subjects aged 18–55 years who were able to provide consent and adhere to the visit schedule and other protocol requirements were eligible for the study. If sexually active and potentially fertile (both males and females), volunteers had to agree to use two forms of contraception (one barrier method) for the duration of the study.

[0278] Exclusion criteria included inappropriate access to the forearm vein, pregnancy or breastfeeding, use of nicotine-containing products during the study, current or recent (within 12 months) history of alcohol or drug abuse, recent (within 90 days) blood donation, and involvement in a clinical trial within the past 90 days. Subjects with excessive somnolence and those who had used medications or drugs that could cause drowsiness within the past 7 days were also excluded. Volunteers were also excluded if their medical history, examination, or clinical laboratory tests indicated significant medical or psychiatric illness that could affect study results or interfere with informed consent and study compliance.

[0279] clinical research design The SAD study (NP10679-101) was a single-center, randomized, double-blind, placebo-controlled, single-dose, dose-escalation trial to investigate the safety, tolerability, and pharmacokinetics (PK) of NP10679 in healthy adult volunteers in six ascending dose cohorts. The primary objective of the study was to assess the safety, tolerability, and PK of a single dose of NP10679 delivered by IV infusion compared with placebo. Secondary objectives were to obtain the maximum tolerated dose of NP10679 in healthy adult volunteers and to establish a safe starting dose for the MAD study (NP10679-102).

[0280] The study consisted of a 30-day screening period, Day 1 (a single IV infusion of randomized NP10679 or placebo), an in-clinic / overnight assessment on Day 2, and an assessment on Day 3. Subjects were admitted to the clinic on Day 1 and stayed there 48 hours after administration until blood collection on Day 3, at which point they were discharged. Subjects returned to the clinic for a follow-up visit on Day 8 after discharge.

[0281] NP10679-101 studied six dose cohorts. Each cohort consisted of eight subjects. Six subjects in each cohort received NP10679 and two subjects received placebo. Doses were sequentially escalated before titrating to the next dose level. Doses included in the study were 5 mg, 15 mg, 50 mg, 100 mg, and 200 mg. Drug and placebo were administered over 30 minutes via IV infusion in 75 ml of dosing vehicle. A sentinel dosing adaptive design approach was used for all cohorts, with the first two subjects (one active, one placebo) dosed on Day 1 and observed for 48 hours or until sufficient time had passed to assess safety. If the safety committee (at a minimum, the principal investigator (PI) and medical monitor (MD in a subject area independent of the conduct of the study) agreed that it was safe to proceed, the remaining six subjects in that cohort (five active, one placebo) were dosed at the same dose level. Prior to dosing of subjects in the next cohort, safety / tolerability data and available PK data were reviewed. Acceptable results of the interim safety / tolerability review prompted enrollment in the next dose cohort.

[0282] The purpose of the MAD study was to evaluate the safety and pharmacokinetics of NP10679 when administered repeatedly until steady state was achieved. Based on the results from the SAD study, it was determined that steady state could be achieved with once-daily dosing for 5 days. Subjects in the MAD study (NP10679-102) were treated in the same manner as subjects in NP10679-101. The study consisted of a 30-day screening period, dosing on days 1–5 (a single IV infusion of 75 ml of randomized NP10679 or placebo over 30 minutes), an in-clinic / overnight assessment on day 6, and a pre-discharge assessment on day 7. Subjects were admitted to the clinic on day 1 and stayed there until 48 hours after dosing, when blood was drawn on day 7, at which point they were discharged. Subjects returned to the clinic for a follow-up visit on day 9. Three dose cohorts of eight subjects each (six drug and two placebo) were recruited, and dosing was performed similarly to NP10679-101. Dose levels included 25 mg, 50 mg, and 100 mg.

[0283] Safety evaluation Safety / tolerability parameters were assessed according to a protocol schedule and included physical examination, infusion site examination, laboratory findings, neuropsychiatric assessment, vital signs, and treatment-emergent adverse events based on subject-reported tolerability. Endpoints also included hematology, chemistry, urinalysis, and a 12-lead ECG. Standard assessments included the Hamilton Depression Rating Scale (HDRS), Mini-Mental State Examination (MMSE), Suicidal Behavior Questionnaire-Revised (SBQ-R), the 7-item Generalized Anxiety Disorder Scale (GAD-7), and the Clinician-Administered Dissociative States Scale (CADSS). The Modified Observer's Assessment of Alertness / Sedation (MOAA / S) and the Bond-Lader VAS sleepiness scale were also included.

[0284] All subjects who received at least one dose of study drug and safety follow-up were included in the safety analysis, regardless of whether they were prematurely discontinued. Data were summarized by reporting the number and percentage of subjects in each category for categorical and ordinal measures, and the mean, SD, median, and range for continuous measures. Safety endpoints included a summary of treatment-emergent clinic- and laboratory-based adverse events and their severity. All adverse events were coded by System Organ Class and Preferred Term according to the Medical Dictionary for Regulatory Affairs (MedDRA). Treatment-emergent adverse events were tabulated by dose level, system organ class, and preferred term.

[0285] Pharmacokinetic measurements For the SAD study, blood samples were collected (when possible) via a vein opposite the infused arm to determine systemic NP10679 levels at pre-dose and at the end of the infusion (20 min ± 5 min), as well as at 0.5, 1, 2, 4, 6, 8, 10, 12, 18, 24, 36, and 48 h post-dose. A 5 mL whole blood sample was collected at each time point using a collection tube containing K2EDTA. Immediately after collection, the tube was inverted to mix the anticoagulant with the blood sample. The tube was then centrifuged at approximately 3000 g for 10 minutes at 4°C. Within 5 minutes of centrifugation, the plasma fraction was transferred in two equal aliquots (1.25 mL each) to 2 mL cryovials, which were then frozen and stored at -70°C (± 10°C) until shipment. For the MAD study, blood was drawn (when possible) via a vein contralateral to the infused arm pre-dose and at the end of the infusion (30 min ± 5 min), as well as at 0.5, 1, 2, 4, 6, 8, 10, 12, and 18 hours on days 1-5, and at 24, 36, 48, and 96 hours after the final dose on day 5, again to determine systemic NP10679 levels.

[0286] A sensitive, specific, and reproducible bioanalytical method was developed and validated at MPI Research (Mattawan, MI) to quantify NP10679. Standards, controls, and test plasma samples containing NP10679 were quantified by a validated LC-MS / MS assay following protein precipitation. A structural analog of NP10679 (NP10767, structure shown below) was used as the internal standard (IS). The method was adapted and used to measure plasma samples by TMD Pharmaceutical Research (Newark, DE). Chromatographic retention of NP10679 and IS was obtained on an Agilent Poroshell 120 EC-C18, 2.1 x 30 mm, 2.7 μm column (Santa Clara, California) under gradient conditions at a flow rate of 0.3 mL / min. Analytes were detected by multiple reaction monitoring using an MDS Sciex API 4000 mass spectrometer (AB Sciex, Framingham, MA) in positive mode. Plasma concentrations from the resulting LC-MS / MS data were calculated using a 6- to 10-point calibration curve constructed from known concentrations of NP10679. The lower limit of quantification (LLOQ) for NP10679 was 2 ng / mL in diluted plasma. [ka]

[0287] Descriptive pharmacokinetic parameters were calculated based on the plasma concentrations of NP10679. Pharmacokinetic analysis was performed based on non-compartmental analytical methods (M. Garibaldi and D Perrier, Pharmacokinetics 2 nd Edition, Chapter 11, Marcel Dekker Inc., New York, 1982), and was performed using MS Excel®.

[0288] B. Results Forty-eight subjects were enrolled in six cohorts of the NP10679-101 study (Table 6), with 47 subjects completing. One subject voluntarily withdrew from the study for personal reasons unrelated to the study. The median age in the study was 33.5 years (youngest / oldest - 22 / 52 years). Thirty men and 18 women were enrolled in the study. Most subjects were black (35), followed by whites (13, including 10 non-Hispanic and 3 Hispanic), and one was Asian. [Table 6]

[0289] The NP10679-102 MAD study enrolled 24 subjects in four cohorts (Table 7). The median age in this study was 44.5 years (youngest / oldest - 20 / 54 years). The study enrolled 15 men and 9 women. Similar to the SAD study, most subjects were Black (15), followed by White (8) and Asian (1). [Table 7]

[0290] Table 8 summarizes treatment-emergent adverse events (TEAEs) by system organ class and dose of NP10679-101. At the highest dose tested, 200 mg, the most common treatment-emergent adverse event (TEAE) was somnolence. [Table 8-1] [Table 8-2]

[0291] There was a dose response for somnolence, the most common TEAE (see Table 9). The Modified Observer Assessment of Alertness / Sedation (MOAA / S) scale was scored from 0 to 5, with level 5 representing the lowest level of sedation. At level 5, the subject responded easily to normal speech, level 4 indicated a lethargic response to voice, and level 3 required a louder voice to elicit a response. A score below 3 required increasing levels of physical stimulation to awaken the subject. NP10679 elicited a moderate effect at higher dose levels on the MOAA / S scale. One of six subjects at the 5 mg and 15 mg doses and five of six subjects at the 50-200 mg doses exhibited somnolence. Two subjects in each of the 100 mg and 150 mg cohorts achieved a transient level 3 score on the MOAA / S scale; however, no subjects in the highest dose group (200 mg) achieved this score. There was also a consistent moderate increase in the Bond-Lader VAS scale, starting with the 50 mg dose and continuing up to the 200 mg dose. The somnolence observed in the study appeared phenomenologically different from that observed with typical sedative-hypnotic drugs. This reduced the overall reliability of the tool used to score somnolence. Even at the highest dose tested, subjects were presented with stimuli and quickly acclimatized to their environment within seconds, allowing them to perform relatively complex tasks, such as the Digit Symbol Substitution Test (DSST). [Table 9]

[0292] At doses of 100 mg or higher, the TEAE of dizziness may also be more frequent. This was reported in one subject at 100 mg and two at 150 mg and 200 mg. Headache and near-syncope were less common and did not appear to demonstrate a dose response. The TEAE of tremor occurred once in a subject receiving 100 mg. None of these TEAEs were considered to affect the safety of the subjects. Outside of nervous system disorders, conjunctival and scleral hyperemia were observed in three of six subjects receiving the 200 mg dose. This was considered related to the clinically insignificant hypotension (both systolic and diastolic) observed at the two highest doses. However, hypotension was more pronounced at 4 hours post-dose with the 150 mg dose of NP10679 (-23 mg Hg) than with the 200 mg dose (-7.2 mg Hg). There were no clinically significant changes in vital signs or ECGs during the study. Thus, the increase in QTc interval or hypertension observed with previous GluN2B inhibitors was not observed in the NP10679-101 study.

[0293] No serious adverse events (SAEs) were observed in the study. There were no patterns suggestive of dissociative symptoms or cognitive impairment associated with NP10679 in this study, as indicated by the Clinician Administered Dissociative Status Scale (CADSS) or Digit Symbol Substitution Test (DSST). At the 150 mg dose, one subject exhibited intrusive thoughts, but there were no patterns suggestive of dissociative symptoms associated with NP10679 in this study, as indicated by the Clinician Administered Dissociative Status Scale (CADSS).

[0294] Table 10 summarizes treatment-emergent adverse events (TEAEs) for NP10679-102 by system organ class and dose. As in the SAD study, there were no SAEs in the MAD study. The most frequently encountered adverse effect, also consistent with the SAD study, was somnolence. This side effect was observed in three subjects in both the 50 mg and 100 mg groups. However, it was also observed in three subjects in the placebo group. There were no signs of increased somnolence upon repeated dosing. Because this effect was observed for the most part only on the first and second days of dosing, there may have been some adaptation to the somnolence effect, but there was not enough pattern to support firm conclusions. There were no patterns suggestive of dissociative symptoms associated with NP10679 in this study, as indicated by the Clinician-Administered Dissociative Status Scale (CADSS). [Table 10-1] [Table 10-2]

[0295] In the NP10679-101 study, NP10679 plasma concentrations (see Figure 5 and Table 11) increased linearly with dose, with mean C max The plasma concentrations ranged from 30.0 ± 14.8 ng / mL at the 5 mg dose to 2066 ± 798 ng / mL at the 200 mg dose. Plasma concentrations then declined multiexponentially, with terminal half-lives ranging from 27.6 ± 12.0 hours to 17.4 ± 2.8 hours, respectively. Systemic clearance ranged from 9.82 ± 2.89 L / h to 10.4 ± 2.51 L / h across the doses studied. NP10679 was eliminated slowly from the body, with less than 12% of hepatic blood flow compared with 87 L / h in humans. NP10679 appeared to be distributed extensively throughout the body, with a volume of distribution exceeding 221 L, equivalent to 4.5 times the total body water space.

[0296] Based on the power model approach, AUC (0~inf)There was a dose-proportional linear increase with NP10679, suggesting that NP10679 follows linear kinetics between 5 and 200 mg. [Table 11]

[0297] In the MAD study of NP10679-102, all subjects except one subject in the 50 mg cohort had quantifiable concentrations of NP10679 in plasma by 24 hours after the first four doses (pre-dose time point on the following day) and by 96 hours after the fifth dose (day 5) (final PK time point). Mean C max increased with increasing dose (Figure 6). Over the doses studied, from 25 to 100 mg, C on days 1 and 5 max increased 5.9-fold and 2.7-fold, respectively (see Table 12 for pharmacokinetic parameters). The mean AUC also increased with increasing dose. At 25-100 mg, the AUC on day 1 0-24h increased 3.8-fold, and the AUC 0-24h and AUC 0-96h The increases were 4.0 and 3.6 times, respectively. max Both RI and AUC were approximately linear with increasing dose. Terminal half-lives were similar across all doses and days studied, with mean ranges of 15.4 to 36.2 hours, 15.5 to 25.6 hours, and 12.5 to 34.0 hours for the 25 mg, 50 mg, and 100 mg cohorts, respectively. Steady-state clearance was similar across doses studied, with means of 11.5 L / h, 11.8 L / h, and 11.2 L / h for the 25 mg, 50 mg, and 100 mg cohorts, respectively. Steady-state volume of distribution decreased slightly with increasing dose, with means of 360 L, 302 L, and 252 L for the 25 mg, 50 mg, and 100 mg cohorts, respectively. [Table 12]

[0298] Adverse events observed in clinical trials were mild and limited to mild somnolence. This appeared to be dose-related, beginning with the intermediate dose of 50 mg in the SAD study. The observed somnolence did not appear to worsen over the 5-day course of administration in the MAD study. The observed somnolence was not similar to that observed with classical sedatives. Even at the highest dose (200 mg) in the SAD study, subjects were easily awakened and able to complete complex tasks, such as the Digit Symbol Substitution Test (DSST).

[0299] Notably, no dissociative symptoms or cognitive decline were observed in either the SAD or MAD studies. Additionally, no clinically significant cardiovascular events were observed.

[0300] Pharmacokinetic data from the SAD and MAD studies indicate linear exposure with dose and a half-life (approximately 20 hours) suitable for once-daily dosing.

[0301] In conclusion, early human studies NP10679-101 and NP10679-102 demonstrate that NP10679 is safe at the doses tested.

Claims

1. Structure of Formula I 【Chemical Formula 28】 or a pharmaceutically acceptable salt, hydrate or hydrate salt thereof [In the formula, The stereocenters marked with an * have the R configuration; R １ teeth, 【Chemical 34】 is selected from R ２ and R ３ are independently selected from hydrogen, methyl, and halomethyl.

2. R １ but, 【Chemistry 30】 2. The compound of claim 1, wherein:

3. R １ but, 【Chemical 31】 2. The compound of claim 1, wherein:

4. R ２ and R ３ The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, hydrate or hydrated salt thereof, wherein

5. 【Chemical 32】 2. The compound of claim 1, or a pharmaceutically acceptable salt, hydrate or hydrate salt thereof, selected from:

6. Structure of Formula II 【Chemical 33】 or a pharmaceutically acceptable salt, hydrate or hydrate salt thereof [In the formula, The stereocenters marked with an * have the R configuration; R ４ is selected from hydrogen, methyl, halomethyl, ethyl, haloethyl, isopropyl, and haloisopropyl; R ５ and R ６ are independently selected from hydrogen, methyl, and halomethyl.

7. R ４ 7. The compound of claim 6, or a pharmaceutically acceptable salt, hydrate or hydrate salt thereof, wherein is selected from methyl and halomethyl.

8. R ５ and R ６ 8. The compound of claim 6 or 7, or a pharmaceutically acceptable salt, hydrate or hydrated salt thereof, wherein:

9. 10. A composition comprising the compound of claim 1, or a pharmaceutically acceptable salt, hydrate or hydrate salt thereof, wherein the R configuration is present in an enantiomeric excess of greater than 80%, greater than 85%, greater than 90%, or greater than 95% with respect to the stereocenter labeled with an * as depicted in Formulas I and II.

10. 10. The composition of claim 9, wherein the compound is present in the R configuration with respect to the stereocenter labeled with an * as depicted in Formulas I and II in an enantiomeric excess of greater than 95%.

11. 11. A pharmaceutical formulation comprising the compound of claim 1, or a pharmaceutically acceptable salt, hydrate or hydrated salt thereof, or the composition of claim 9 or 10, wherein the pharmaceutical formulation further comprises a pharmaceutically acceptable excipient.

12. 12. The pharmaceutical formulation of claim 11, wherein the pharmaceutical formulation is present in a form selected from a tablet, a capsule, a caplet, a pill, a bead, a granule, a fine granule, a powder, a gel, a cream, a liquid, a suspension, an emulsion, and a nanoparticle formulation.

13. 12. The pharmaceutical formulation of claim 11, wherein the pharmaceutical formulation is an oral or intravenous formulation.

14. 12. The pharmaceutical formulation of claim 11, wherein the pharmaceutical formulation is in the form of a lyophilized powder.

15. 12. The pharmaceutical formulation of claim 11, wherein the pharmaceutical formulation is in the form of a sterile aqueous solution.

16. 11. A composition comprising a compound of claim 1 or a pharmaceutically acceptable salt, hydrate or hydrate salt thereof, or a composition of claim 9 or 10, for treating a condition, disorder or disease in a subject in need thereof, The composition, wherein the condition, disorder or disease is selected from stroke, subarachnoid hemorrhage, cerebral ischemia, cerebral vasospasm, hypoxia, acute CNS injury, spinal cord injury, traumatic brain injury, coronary artery bypass graft, persistent or chronic cough, substance abuse disorder, opiate withdrawal, opiate tolerance, bipolar disorder, suicidal ideation, pain, fibromyalgia, depression, postpartum depression, resting tremor, dementia, epilepsy, seizure disorders, movement disorders, and neurodegenerative diseases.

17. 17. The composition of claim 16, wherein the condition, disorder or disease is pain, depression, stroke, or subarachnoid hemorrhage.

18. 17. The composition of claim 16, wherein the pain is neuropathic pain.

19. 17. The composition of claim 16, wherein the pain is chronic pain.

20. The composition of claim 16, wherein the pain is cancer pain.

21. 17. The composition of claim 16, wherein the depression is treatment-resistant depression.

22. 17. The composition of claim 16, wherein the neurodegenerative disease is Huntington's disease, Alzheimer's disease, or Parkinson's disease.

23. 17. The composition of claim 16, wherein the epilepsy is caused by a genetic mutation.

24. 17. The composition of claim 16, wherein the seizure disorder is infantile spasticity.

25. 17. The composition of claim 16, wherein the dementia is AIDS-induced dementia.

26. 17. The composition of claim 16, wherein the hypoxia is induced by respiratory failure, prolonged use of a ventilator, or both.

27. 27. The composition of claim 26, wherein the respiratory failure, prolonged use of a ventilator, or both, is associated with COVID-19.

28. 17. The composition of claim 16, wherein the composition is administered orally or intravenously.

29. The composition of claim 16 , wherein the subject is a human.

30. A composition comprising the compound of claim 6, or a pharmaceutically acceptable salt, hydrate or hydrated salt thereof, wherein the compound is in the R configuration with respect to the stereocenter labeled with an * as depicted in Formulas I and II, in an enantiomeric excess of greater than 80%, greater than 85%, greater than 90%, or greater than 95%.

31. The composition of claim 30, wherein the compound is present in the R configuration with respect to the stereocenter labeled with an * as depicted in Formulas I and II in an enantiomeric excess of greater than 95%.

32. A pharmaceutical formulation comprising the compound of claim 6, or a pharmaceutically acceptable salt, hydrate or hydrated salt thereof, or the composition of claim 30 or 31, wherein the pharmaceutical formulation further comprises a pharmaceutically acceptable excipient.

33. The pharmaceutical formulation of claim 32, wherein the pharmaceutical formulation is present in a form selected from tablets, capsules, caplets, pills, beads, granules, fine granules, powders, gels, creams, liquids, suspensions, emulsions, and nanoparticle formulations.

34. The pharmaceutical formulation of claim 32, wherein the pharmaceutical formulation is an oral or intravenous formulation.

35. The pharmaceutical formulation of claim 32, wherein the pharmaceutical formulation is in the form of a freeze-dried powder.

36. The pharmaceutical formulation of claim 32, wherein the pharmaceutical formulation is in the form of a sterile aqueous solution.

37. A composition comprising the compound of claim 6, or a pharmaceutically acceptable salt, hydrate, or hydrate salt thereof, or the composition of claim 30 or 31, for treating a condition, disorder, or disease in a subject in need thereof, comprising: The composition, wherein the condition, disorder or disease is selected from stroke, subarachnoid hemorrhage, cerebral ischemia, cerebral vasospasm, hypoxia, acute CNS injury, spinal cord injury, traumatic brain injury, coronary artery bypass graft, persistent or chronic cough, substance abuse disorder, opiate withdrawal, opiate tolerance, bipolar disorder, suicidal ideation, pain, fibromyalgia, depression, postpartum depression, resting tremor, dementia, epilepsy, seizure disorders, movement disorders, and neurodegenerative diseases.

38. The composition described in claim 37, wherein the condition, disorder or disease is pain, depression, stroke, or subarachnoid hemorrhage.

39. The composition described in claim 37, wherein the pain is neuropathic pain.

40. The composition described in claim 37, wherein the pain is chronic pain.

41. The composition described in claim 37, wherein the pain is cancer pain.

42. The composition described in claim 37, wherein the depression is treatment-resistant depression.

43. The composition described in claim 37, wherein the neurodegenerative disease is Huntington's disease, Alzheimer's disease, or Parkinson's disease.

44. The composition described in claim 37, wherein the epilepsy is caused by a genetic mutation.

45. The composition described in claim 37, wherein the seizure disorder is infantile spasticity.

46. The composition described in claim 37, wherein the dementia is AIDS-induced dementia.

47. The composition described in claim 37, wherein the hypoxia is induced by respiratory failure, prolonged use of a ventilator, or both.

48. The composition of claim 47, wherein the respiratory failure, prolonged use of a ventilator, or both, is associated with COVID-19.

49. The composition described in claim 37, wherein the composition is administered orally or intravenously.

50. The composition described in claim 37, wherein the subject is a human.

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