Therapeutically Effective Oral Administration of 2-Arylbenzimidazoles

Abstract

A method for reducing neuroinflammation and / or treating a neurodegenerative disease in a subject is provided, the method comprising administering to a subject having a neuroinflammation and / or neurodegenerative disease a compound of formula (I): The method includes orally administering a pharmaceutical composition comprising TIFF2024522820000066.tif20146 (TQS-168), or a pharma- ceutically acceptable salt thereof, in an amount that results in a predefined plasma and brain exposure of TQS-168 and / or an active metabolite after administration. In various embodiments, the TQS-168 or a salt thereof is administered in an amount that results in a mean plasma Cmax of TQS-168 of at least 1000 ng / mL, at least 1250 ng / mL, at least 1500 ng / mL, or at least 1750 ng / mL after administration.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of and priority to U.S. Provisional Application No. 63 / 211,636, filed June 17, 2021, and U.S. Provisional Application No. 63 / 300,551, filed January 18, 2022, each of which is incorporated by reference in its entirety for all purposes. [Background technology]

[0002] 1. Background of the Invention The 2-arylbenzimidazole compound TQS-168 (2-(4-tert-butylphenyl)-1H-benzimidazole), formerly known as ZLN-005, is an activator of Ppargc1α (PGC-1α) gene expression. Zhang et al., Diabetes 62:1297-1307 (2013). When administered orally to mice at 25-50 mg / kg, TQS-168 has been shown to suppress bone marrow-mediated inflammation and reduce disease severity in mouse models of neurodegenerative diseases including Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis (ALS). See U.S. Patent No. 10,272,070. When orally administered to mice at 25 mg / kg, TQS-168 has also been shown to suppress metabolic dysfunction in microglial cells in aged mice, inhibit inflammatory cytokine production in microglial cells in aged mice, suppress systemic inflammation in aged mice, and alleviate behavioral dysfunction in aged mice. See U.S. Patent No. 10,653,669. TQS-168 and structurally related 2-arylbenzimidazoles have also been shown to be effective in treating systemic immune activation. See WO2021 / 262617. TQS-168 is highly insoluble. In the animal model studies reported in U.S. Patents 10,272,070 and 10,653,669, TQS-168 was prepared as an oral suspension and administered to experimental animals by oral gavage. Plasma and brain concentrations of the compound after administration were not reported, and no pharmacokinetic (PK) information was provided. In order to establish an effective oral dosing regimen suitable for human patients, it is necessary to define the plasma and brain concentrations and total exposure of TQS-168 that result in pharmacodynamic benefit. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] U.S. Patent No. 10,272,070 [Patent Document 2] U.S. Pat. No. 10,653,669 [Patent Document 3] International Publication No. 2021 / 262617 [Non-patent literature]

[0004] [Non-Patent Document 1] Zhang et al., Diabetes 62:1297-1307 (2013). Summary of the Invention

[0005] 2. Summary of the Invention We now demonstrate that TQS-168 induces PGC-1α protein expression in vitro in a murine myeloid cell line, BV2, at concentrations ranging from 0.7 μM (175.21 ng / mL) to 20 μM (5006 ng / mL), and that TQS-168 inhibits LPS-induced secretion of proinflammatory cytokines from BV2 cells and human primary myeloid cells in vitro at concentrations ranging from 0.3 μM (75.09 ng / mL) to 20 μM (5006 ng / mL).

[0006] These in vitro experiments predict that plasma free drug and brain concentrations of TQS-168 in the range of 0.3-20 μM (75.09-5006 ng / mL) should suppress bone marrow-mediated neuroinflammation.

[0007] When administered orally at 25-50 mg / kg, TQS-168 has previously been shown to suppress bone marrow-mediated inflammation and reduce disease severity in mouse models of neurodegenerative diseases including Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis (ALS). U.S. Patent No. 10,272,070. We are now measuring plasma, liver, and brain concentrations of TQS-168 following administration of a single oral dose of 25 mg / kg to mice, a dose previously found to produce a therapeutic effect. At this previously established effective dose, the mean plasma C of TQS-168 was 1.2-fold higher than that of TQS-168 in mice. max was 93.4ng / ml, or 0.37μM, and the mean brain C max was higher at 542.0 ng / ml, or 2.16 μM. These concentrations have been shown to induce PGC-1α protein expression and reduce LPS-mediated inflammatory cytokine release in vitro within that concentration range. These in vivo data confirm that plasma concentrations of TQS-168 in the range of 0.3 μM to 20 μM should suppress bone marrow-mediated neuroinflammation. Evidence of accumulation in the brain suggests that plasma concentrations of TQS-168 below 0.37 μM may also be effective in treating neuroinflammation.

[0008] We also demonstrate that TQS-621, an early phase 1 metabolite of TQS-168, potently inhibits LPS-induced IL-6 and TNFα secretion from primary human PBMCs. These data demonstrate that at least some of the therapeutic effects observed after oral administration of TQS-168 may be attributable to the activity of the metabolite TQS-621.

[0009] The inventors have also conducted a Phase 1 human clinical trial measuring plasma concentrations of TQS-168 and the active metabolite TQS-621 in three different formulations to demonstrate that pharmacodynamically relevant plasma concentrations can be achieved in oral suspensions of several solid formulations of the API.

[0010] Thus, in a first aspect, a method is provided for reducing neuroinflammation and / or treating a neurodegenerative disease in a subject, the method comprising: A subject having a neuroinflammatory and / or neurogenerative disorder is administered a compound of formula (I) [ka] (TQS-168), or a pharma- ceutical acceptable salt thereof, A mean peak concentration (C) of TQS-168 in plasma of at least 750 ng / mL max ) This includes orally administering in an amount that results in

[0011] In various embodiments, TQS-168 or a salt thereof is administered in an amount that results in a mean plasma Cmax of TQS-168 after administration of at least 1000 ng / mL, at least 1250 ng / mL, at least 1500 ng / mL, or at least 1750 ng / mL.

[0012] In some embodiments, TQS-168 or a salt thereof is administered at a concentration of at least 3000 ng·hr / ml, at least 4000 ng·hr / ml, at least 5000 ng·hr / ml, at least 5500 ng·hr / ml, at least 6000 ng·hr / ml, or at least 7000 ng·hr / ml after administration. * hr / ml, or at least 7,000ng * AUC in hr / ml 0-t In certain embodiments, the TQS-168 or salt thereof is administered in an amount that results in an AUC of about 6000 ng·hr / ml after administration. 0-t The compound is administered in an amount that results in

[0013] In various embodiments, the time to plasma Cmax (Tmax) of TQS-168 is 2 hours or less, 90 minutes or less, or 75 minutes or less. In certain embodiments, the TQS-168 plasma Tmax is about 60 minutes.

[0014] In a second aspect, a method is provided for reducing neuroinflammation and / or treating a neurodegenerative disease in a human subject, the method comprising: A subject having a neuroinflammatory and / or neurogenerative disorder is administered a compound of formula (I) [ka] (TQS-168), or a pharma- ceutical composition comprising ... At least 1000 ng / mL of the compound of formula (II) [ka] The mean peak plasma concentration (C max ) This includes orally administering in an amount that results in

[0015] In some embodiments, TQS-168 or a salt thereof is administered to a subject in need of a medicament for which the plasma C of TQS-621 is between 200 and 2750 ng / mL, between 300 and 2200 ng / mL, or between 400 and 1800 ng / mL. max The compound is administered in an amount that results in

[0016] In another embodiment, a method is provided for reducing neuroinflammation and / or treating a neurodegenerative disease in a subject, the method comprising: A subject having a neuroinflammatory and / or neurogenerative disorder is administered a compound of formula (I) [ka] (TQS-168), or a pharma- ceutical composition comprising ... (a) A mean peak concentration (C) of TQS-168 in plasma of at least 750 ng / mL max )of, (b) C of TQS-168 in plasma at 75 min or less max Average time to max ); and (c) at least 1000 ng / mL of the compound of formula (II) in plasma [ka] (TQS-621) average peak concentration (C max ), (d) C of TQS-621 in plasma at 4 hours or less max Average time to max ) This includes orally administering in an amount that results in

[0017] In some embodiments of the methods herein, TQS-168, or a salt thereof, is administered in a daily oral dose of 200-800 mg, 300-700 mg, 400-600 mg, or 400-500 mg. In certain embodiments, TQS-168, or a salt thereof, is administered in a daily oral dose of 400 mg or 450 mg.

[0018] In various embodiments of the methods described herein, TQS-168 or a salt thereof is administered in a liquid suspension. In certain embodiments, TQS-168 or a salt thereof is administered in a liquid solution.

[0019] In certain embodiments, TQS-168 or its salt is administered in solid dosage form. In certain solid form embodiments, TQS-168 or its salt is crystalline. In certain solid form embodiments, TQS-168 or its salt is non-crystalline, and in particular non-crystalline embodiments, it is a spray-dried dispersion or a hot melt extrudate. In certain embodiments, the solid dosage form is a sachet, capsule, or tablet.

[0020] In various embodiments, the subject has a neurodegenerative disease selected from motor neuron disease, amyotrophic lateral sclerosis (ALS), Alzheimer's disease, vascular dementia, frontotemporal degeneration (frontotemporal dementia), dementia with Lewy bodies, Parkinson's disease, Huntington's disease, demyelinating disease, and multiple sclerosis (MS).In certain embodiments, the subject has a motor neuron disease.In specific embodiments, the subject has ALS.In certain embodiments, the subject has Alzheimer's disease.

[0021] In some embodiments, the subject is at least 40 years of age and has no previously diagnosed neurodegenerative disease. In certain embodiments, the subject is at least 60 years of age or at least 65 years of age.

[0022] 3. Brief description of the drawings These and other features, aspects, and advantages of the present invention will become better understood with regard to the following description and accompanying drawings: [Brief description of the drawings]

[0023] [Figure 1] FIG. 1 is a Western blot showing induction of PGC-1α protein expression in the BV2 mouse microglial cell line following incubation with TQS-168 at 20 μM in vitro.

[0024] [Diagram 2] FIG. 2 is a Western blot showing the dose response of increased PGC-1α protein expression in BV2 cells contacted with TQS-168 in vitro.

[0025] [Diagram 3] Figure 3 is a bar graph quantifying protein expression levels measured from scans of the Western blots shown in Figure 2. "PD" refers to PD169316, a p38 MAPK inhibitor. "Rosi" refers to rosiglitazone (AVANDIA), a PPARγ agonist.

[0026] [Figure 4a-4f] Figures 4a-4f show in vitro secretion of cytokines by BV2 cells after LPS activation in the presence of two positive controls, one negative control, and four different concentrations of TQS-168. The graphs show dose-response inhibition of LPS-stimulated TNFα and IL-6 cytokine release by various concentrations of TQS-168. Cytokine secretion was measured using a cytokine bead array (CBA) fluorescent activated cell sorting (FACS) assay.

[0027] [Figure 5a-5b] 5a-5b show inhibition of TNFα production by LPS-stimulated BV2 bone marrow cells in vitro by treatment with 5 μM (FIG. 5a) and 20 μM (FIG. 5b), respectively, of TQS-168.

[0028] [Figure 6a-6d] Figures 6a-6d show the dose response of TQS-168-mediated inhibition of the proinflammatory cytokine TNFα release from LPS-stimulated microglial BV2 cells at 24 hours using ELISA. Figures 6a and 6c show the absolute (Figure 6a) and relative (Figure 6c) inhibition of TNFα secretion by BV2 cells stimulated with 0.3 ng / mL of LPS. Figures 6b and 6d show the absolute (Figure 6b) and relative (Figure 6d) inhibition of TNFα secretion by BV2 cells stimulated with 1 ng / mL of LPS.

[0029] [Figure 7] FIG. 7 shows the inhibition of LPS-induced TNFα secretion in human PBMC cells treated in vitro with various concentrations of TQS-168.

[0030] [Figure 8a-8c] 8a-8c show the mean plasma (FIG. 8a), liver (FIG. 8b), and brain (FIG. 8c) concentrations of TQS-168 at various time points following a single oral dose of 25 mg / kg TQS-168 in mice.

[0031] [Figure 9a-9c] 9a-9c show the plasma concentrations of TQS-168 at various time points following a single oral dose of 50 mg / kg in wild type mice.

[0032] [Figure 10] FIG. 10 shows the plasma concentrations of TQS-168 at various time points following a single intravenous (IV) dose of 0.5 mg / kg in three individual mice.

[0033] [Figure 11] FIG. 11 shows the mean plasma concentrations of TQS-168 at various time points following a single intravenous (IV) dose of 0.5 mg / kg in three mice.

[0034] [Figure 12A-12B] Figure 12a shows the plasma concentration of TQS-168 at various time points after a single oral dose of 50, 150, or 500 mg / kg in rats, and Figure 12b shows the mean brain concentration of TQS-168 at various time points after a single oral dose of 500 mg / kg in rats.

[0035] [Figure 13] FIG. 13 shows the dose-dependent Cmax (ng / mL) values ​​of TQS-168 following oral administration of 50 mg / kg, 150 mg / kg, and 500 mg / kg in rats.

[0036] [Figure 14] FIG. 14 shows the dose-dependent AUC (ng*min / mL) of TQS-168 following oral administration of 50 mg / kg, 150 mg / kg, and 500 mg / kg in rats.

[0037] [Figure 15] FIG. 15 shows the plasma concentrations of TQS-168 at various time points following a single intravenous (IV) dose of 0.5 mg / kg in three rats.

[0038] [Figure 16]FIG. 16 shows the mean plasma concentrations of TQS-168 at various time points following a single intravenous (IV) dose of 0.5 mg / kg in rats.

[0039] [Figure 17] FIG. 17 shows the plasma concentrations of TQS-168 at various time points following a single intravenous (IV) dose of 0.5 mg / kg in three dogs.

[0040] [Figure 18] FIG. 18 shows the mean plasma concentrations of TQS-168 at various time points following a single intravenous (IV) dose of 0.5 mg / kg in dogs.

[0041] [Figure 19A] FIG. 19A shows the mean plasma concentrations of TQS-168 following oral dosing of 45 mg / kg TQS-168 in mice. [Figure 19B] FIG. 19B shows the mean brain concentrations of TQS-168 following oral administration of 45 mg / kg TQS-168 in mice.

[0042] [Figure 20] FIG. 20 illustrates the phase 1 metabolites from the hepatic metabolism of TQS-168 following oral administration.

[0043] [Figure 21A] FIG. 21A shows absolute inhibition of LPS-stimulated IL-6 secretion by the TQS-168 metabolite TQS-621 from previously frozen PBMC obtained from the first healthy human volunteer donor. [Figure 21B] FIG. 21B shows the relative inhibition of IL-6 inhibition expressed as percentage activity.

[0044] [Figure 22A] FIG. 22A shows absolute inhibition of LPS-stimulated IL-6 secretion by the TQS-168 metabolite TQS-621 from previously frozen PBMC obtained from a second healthy human donor. [Figure 22B]FIG. 22B shows the relative inhibition of IL-6 inhibition expressed as percentage activity.

[0045] [Figure 23A] FIG. 23A shows absolute inhibition of LPS-stimulated TNFα secretion by TQS-168 and metabolite TQS-621 from previously frozen PBMC of the first donor. [Figure 23B] FIG. 23B shows the relative inhibition of IL-6 inhibition expressed as percentage activity.

[0046] [Figure 24A] FIG. 24A shows absolute inhibition of LPS-stimulated TNFα secretion by TQS-168 and metabolite TQS-621 from previously frozen PBMC obtained from a second donor. [Figure 24B] FIG. 24B shows the relative inhibition of TNFα inhibition expressed as percentage activity.

[0047] [Figure 25A] 25A-25C plot the plasma concentration of TQS-168 over time following a single oral dose of TQS-168 in mice administered 50 mg / mL TQS-168 in different formulations. [Figure 25B] 25A-25C plot the plasma concentration of TQS-168 over time following a single oral dose of TQS-168 in mice administered 50 mg / mL TQS-168 in different formulations. [Figure 25C] 25A-25C plot the plasma concentration of TQS-168 over time following a single oral dose of TQS-168 in mice administered 50 mg / mL TQS-168 in different formulations.

[0048] [Figure 26A] Figures 26A-26C plot the plasma concentration of TQS-621 over time after a single oral dose in mice dosed with different amounts of TQS-168 50 mg / mL. [Figure 26B]Figures 26A-26C plot the plasma concentration of TQS-621 over time after a single oral dose in mice dosed with different amounts of TQS-168 50 mg / mL. [Figure 26C] Figures 26A-26C plot the plasma concentration of TQS-621 over time after a single oral dose in mice dosed with different amounts of TQS-168 50 mg / mL.

[0049] [Figure 27A-27B] Figures 27A-B plot the plasma concentration of TQS-168 over time following a single oral dose of 60 mg, 180 mg, or 540 mg TQS-168 methylcellulose powder suspension formulation in humans. Figure 27A is a linear plot. Figure 27B is a logarithmic plot.

[0050] [Fig. 28A-28B] Figures 28A-B plot the plasma concentration of metabolite TQS-621 over time following a single oral dose of 60 mg, 180 mg, or 540 mg suspension formulations of TQS-168 methylcellulose powder in humans. Figure 28A is a linear plot. Figure 28B is a logarithmic plot.

[0051] [Figure 29A-29B] Figures 29A-B plot the plasma concentrations of TQS-168 and metabolite TQS-621 over time following a single oral dose of a 60 mg TQS-168 methylcellulose powder suspension formulation in humans. Figure 29A is a linear plot. Figure 29B is a logarithmic plot.

[0052] [Fig. 30A-30B] Figures 30A-B plot the plasma concentrations of TQS-168 and metabolite TQS-621 over time following a single oral dose of a 180 mg suspension formulation of TQS-168 methylcellulose powder in humans. Figure 30A is a linear plot. Figure 30B is a logarithmic plot.

[0053] [Fig. 31A-31B]Figures 31A-B plot the plasma concentrations of TQS-168 and metabolite TQS-621 over time following a single oral dose of a 540 mg suspension oral formulation of TQS-168 methylcellulose powder in humans. Figure 31A is a linear plot. Figure 31B is a logarithmic plot.

[0054] [Fig. 32A-32B] Figures 32A-B plot the plasma concentration of TQS-168 over time following a single dose of 60 mg, 180 mg, or 540 mg TQS-168 methylcellulose (MC) powder for oral suspension in the fasted state, 90 mg spray dried dispersion (SDD) powder for oral suspension in the fed state, 180 mg SDD powder for oral suspension in the fasted state, or 180 mg hot melt extrudate (HME) powder in the fasted state. Figure 32A is a linear plot. Figure 32B is a logarithmic plot.

[0055] [Fig. 33A-33B] Figures 33A-B plot the plasma concentration of metabolite TQS-621 over time following a single dose of 60 mg, 180 mg, or 540 mg TQS-168 methylcellulose (MC) powder for oral suspension in the fasted state, 90 mg spray dried dispersion (SDD) powder for oral suspension in the fed state, 180 mg SDD powder for oral suspension in the fasted state, or 180 mg hot melt extrudate (HME) powder in the fasted state. Figure 33A is a linear plot. Figure 33B is a logarithmic plot.

[0056] [Fig. 34A-34B] Figures 34A-B plot the plasma concentrations of TQS-168 and metabolite TQS-621 over time following a single dose of a 90 mg suspension of TQS-168 spray-dried dispersion (SDD) powder (oral formulation) in fed-state humans. Figure 34A is a linear plot. Figure 34B is a logarithmic plot.

[0057] [Fig. 35A-35B]Figures 35A-B plot the plasma concentration of TQS-168 over time following a single dose of a 90 mg suspension of spray-dried dispersion (SDD) powder (oral formulation) in humans in fed and fasted states. Figure 35A is a linear plot. Figure 35B is a logarithmic plot.

[0058] [Fig. 36A-36B] Figures 36A-B plot the plasma concentration of metabolite TQS-621 over time following a single dose of a 90 mg suspension of spray-dried dispersion (SDD) powder (oral formulation) in humans in fed and fasted states. Figure 35A is a linear plot. Figure 35B is a logarithmic plot.

[0059] [Fig. 37A-37B] Figures 37A-B plot the plasma concentrations of TQS-168 and metabolite TQS-621 over time following a single oral dose of a 90 mg suspension of TQS-168 spray-dried dispersion (SDD) powder (oral formulation) in fasted humans. Figure 37A is a linear plot. Figure 37B is a logarithmic plot.

[0060] [Fig. 38A-38B] Figures 38A-B plot the plasma concentration of TQS-168 over time following a single dose of 90 mg, 180 mg, or 270 mg suspension of TQS-168 spray dried dispersion (SDD) powder (oral formulation) in fasted humans. Figure 38A is a linear plot. Figure 38B is a logarithmic plot.

[0061] [Figure 39A-39B] Figures 39A-B plot the plasma concentration of metabolite TQS-621 over time following a single dose of 90 mg, 180 mg, or 270 mg suspension of TQS-168 spray dried dispersion (SDD) powder (oral formulation) in fasted humans. Figure 39A illustrates a linear plot. Figure 39B illustrates a logarithmic plot.

[0062] [Fig. 40A-40B]Figures 40A-B plot the plasma concentration of TQS-168 over time following a single dose of 90 mg spray dried dispersion (SDD) powder or 120 mg suspension (oral formulation) in humans in the fed and fasted states, respectively (Day 1). Figure 40A is a linear plot. Figure 40B is a logarithmic plot.

[0063] [Figure 41A-41B] Figures 41A-B plot the plasma concentrations of TQS-168 over time following once daily administration of 90 mg spray dried dispersion (SDD) powder or 120 mg suspension (oral formulation) for 7 consecutive days in humans in fed and fasted states, respectively. Figure 41A is a linear plot. Figure 41B is a logarithmic plot.

[0064] [Fig. 42A-42B] Figures 42A-B plot the plasma concentration of metabolite TQS-621 over time following a single dose of TQS-168 spray dried dispersion (SDD) powder 90 mg or 120 mg suspension (oral formulation) in humans in the fed and fasted states, respectively. Figure 42A is a linear plot. Figure 42B is a logarithmic plot.

[0065] [Fig. 43A-43B] Figures 43A-B plot the plasma concentration of metabolite TQS-621 over time following once daily administration of 90 mg or 120 mg suspension of TQS-621 spray-dried dispersion powder (oral formulation) for 7 consecutive days in humans in the fed and fasted states, respectively. Figure 43A is a linear plot. Figure 43B is a logarithmic plot.

[0066] [Fig. 44A-44B] Figures 44A-B plot the plasma concentration of TQS-168 over time following single administration of various doses of TQS-168 methylcellulose (MC), spray dried dispersion (SDD), or hot melt extrudate (HME) powder suspension formulations in humans. Figure 44A is a linear plot. Figure 44B is a logarithmic plot.

[0067] [Fig. 45A-45B] Figures 45A-B plot the plasma concentration of TQS-621 over time following single dose administration of TQS-168 methylcellulose (MC), spray dried dispersion (SDD), or hot melt extrudate (HME) powder suspension formulations at various doses. Figure 45A is a linear plot. Figure 45B is a logarithmic plot.

[0068] [Fig. 46A-46B] Figures 46A-B plot the plasma concentrations of TQS-168 and metabolite TQS-621 over time following a single dose of a 180 mg suspension of TQS-168 spray dried dispersion (SDD) powder (oral formulation) in fasted humans. Figure 46A is a linear plot. Figure 46B is a logarithmic plot.

[0069] [Fig. 47A-47B] Figures 47A-B plot the plasma concentrations of TQS-168 and metabolite TQS-621 over time following a single dose of a 180 mg suspension of TQS-168 hot melt extrudate (HME) powder (oral formulation) in fasted humans. Figure 34A is a linear plot. Figure 34B is a logarithmic plot.

[0070] [Fig. 48A-48B] Figures 48A-B plot the plasma concentration of TQS-621 over time following single dose administration of TQS-168 methylcellulose (MC), spray dried dispersion (SDD), or hot melt extrudate (HME) powder suspension formulations at various doses in fasted humans. Figure 48A is a linear plot. Figure 48B is a logarithmic plot.

[0071] [Fig. 49A-49B] Figures 49A-B plot the plasma concentration of TQS-168 over time following successive once-daily administration of a 120 mg suspension of the spray-dried dispersion (SDD) powder (oral formulation) in fasted humans. Figure 49A is a linear plot. Figure 49B is a logarithmic plot.

[0072] [Fig. 50A-50B] Figures 50A-B plot the plasma concentration of metabolite TQS-621 over time following successive once-daily administration of a 120 mg suspension of TQS-168 spray-dried dispersion (SDD) powder (oral formulation) in fasted humans. Figure 50A is a linear plot. Figure 50B is a logarithmic plot.

[0073] [Fig. 51A-51B] Figures 51A-B plot the plasma concentration of TQS-168 over time following a single dose of 90 mg, 120 mg, or 300 mg suspension of spray dried dispersion (SDD) powder (oral formulation) in fed / fasted humans (Day 1). Figure 51A is a linear plot. Figure 51B is a logarithmic plot.

[0074] [Fig. 52A-52B] Figures 52A-B plot the plasma concentrations of TQS-168 over time following once daily administration of 90 mg, 120 mg, or 300 mg suspension of spray dried dispersion (SDD) powder (oral formulation) in fed / fasted humans, respectively, for 7 consecutive days. Figure 52A is a linear plot. Figure 52B is a logarithmic plot.

[0075] [Fig. 53A-53B] Figures 53A-B plot the plasma concentration of metabolite TQS-621 over time following a single dose of 90 mg, 120 mg, or 300 mg suspension of spray dried dispersion (SDD) powder (oral formulation) in fed / fasted humans (Day 1). Figure 53A is a linear plot. Figure 53B is a logarithmic plot.

[0076] [Fig. 54A-54B]Figures 54A-B plot the plasma concentration of metabolite TQS-621 over time following single dose administration of 90 mg, 120 mg or 300 mg suspension of TQS-168 spray dried dispersion (SDD) powder (oral formulation) on 7 consecutive days in humans in the fed / fasted state (Day 1). Figure 54A is a linear plot. Figure 54B is a logarithmic plot.

[0077] [Fig. 55A-55B] Figures 55A-B plot the plasma concentration of TQS-168 over time following single administration of various doses of TQS-168 methylcellulose (MC), spray dried dispersion (SDD), or hot melt extrudate (HME) powder suspension formulations. Figure 55A is a linear plot. Figure 55B is a logarithmic plot. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0078] 4. Detailed Description of the Invention 4.1. Definition 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 invention belongs.

[0079] The terms "individual", "host" and "subject" are used interchangeably and refer to the animal to be treated, including, but not limited to, humans and non-human primates; rodents, including rats and mice; cattle; horses; sheep; cats; and dogs. "Mammal" refers to one or more members of any mammalian species. Non-human animal models, i.e., mammals, non-human primates, mice, rabbits, etc., can be used for experimental investigations.

[0080] "Patient" refers to a human subject, including a healthy human donor.

[0081] The terms "treating", "treatment" and grammatical variations thereof are used in the broadest sense as understood in the clinical field. Thus, these terms do not require a cure or complete remission of a disease, but encompass obtaining any clinically desired pharmacological and / or physiological effect. Unless otherwise specified, "treating" and "treatment" do not encompass prophylaxis.

[0082] The phrase "therapeutically effective amount" refers to the amount of a compound that, when administered to a mammal or other subject for treating a disease, condition, or disorder, is sufficient to effect treatment of the disease, condition, or disorder. A "therapeutically effective amount" can vary depending on the compound, the disease and its severity, and the age, weight, etc., of the subject being treated.

[0083] The term "pharmaceutically acceptable salt" refers to a salt that is acceptable for administration to a subject. Examples of pharma- ceutically acceptable salts include, but are not limited to, acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, palmoate, pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, and undecanoate.

[0084] Other examples of pharmaceutical salts include N + , NH4 + , and NW4 -(wherein W can be a C1-C8 alkyl group), and anions of the compounds of the present disclosure in complex with the same. For therapeutic applications, salts of the compounds of the present disclosure may be pharma-ceutically acceptable. However, salts of acids and bases that are not pharma-ceutically acceptable may also be used, for example, in the preparation or purification of a pharma-ceutically acceptable compound.

[0085] Compounds included in the present compositions and methods that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharma- ceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, including, but not limited to, malate, oxalate, chloride, bromide, iodide, nitrate, sulfate, hydrogen sulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts.

[0086] Compounds included in the present compositions and methods that are acidic in nature can form base salts with various pharmacologically acceptable cations. Examples of such salts include the alkali metal or alkaline earth metal salts, particularly the calcium, magnesium, sodium, lithium, zinc, potassium, and iron salts.

[0087] The compounds included in the compositions and methods that contain basic or acidic moieties can also form pharmaceutically acceptable salts with various amino acids.The compounds of the present disclosure can contain both acidic and basic groups; for example, one amino and one carboxylic acid group.In such cases, the compounds can exist as acid addition salts, zwitterions, or base salts.

[0088] Ranges: Throughout this disclosure, various aspects of the invention are expressed in range format. The ranges include the recited endpoints. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as a rigid limitation on the scope of the invention. Thus, the description of a range should be interpreted as having all possible subranges specifically disclosed, as well as individual numerical values ​​within that range. For example, the description of a range such as 1-6 should be interpreted as having specifically disclosed subranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, as well as individual numerical values ​​within that range, e.g., 1, 2, 3, 4, 5, 5.3, and 6. This is true regardless of the breadth of the range.

[0089] In this disclosure, the words "comprises," "comprising," "containing," "having," "includes," "including," and linguistic variants thereof have the meanings given to them in U.S. patent law and permit the presence of additional ingredients beyond those specifically recited.

[0090] Unless specifically stated otherwise or clear from the context, the term "or" as used herein is understood to be inclusive.

[0091] Unless specifically stated otherwise or clear from the context, as used herein, the terms "a," "an," and "the" are understood to be singular or plural. That is, the articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0092] Unless specifically stated or clear from the context, the term "about" as used herein is understood to be within normal tolerances in the art, e.g., within 2 standard deviations from the mean, and is meant to encompass variations of ±20% or ±10%, more preferably ±5%, even more preferably ±1%, and even more preferably ±0.1% from the stated value. When a percentage is presented in relation to the amount of an ingredient or material in a composition, the percentage should be understood to be a percentage by weight unless otherwise stated or understood from the context.

[0093] It should be understood that the order of steps or order for performing certain actions is immaterial so long as the disclosure remains operable. Moreover, two or more steps or actions may be performed simultaneously.

[0094] The terms "pharmaceutically acceptable excipient," "pharmaceutically acceptable diluent," "pharmaceutically acceptable carrier," and "pharmaceutically acceptable adjuvant" are used interchangeably and refer to an excipient, diluent, carrier, or adjuvant that is generally safe, non-toxic, and not biologically or otherwise undesirable and is useful in the preparation of a pharmaceutical composition, and include excipients, diluents, carriers, and adjuvants that are acceptable for veterinary use, as well as human pharmaceutical use. The phrase "pharmaceutically acceptable excipient" includes both one and more than one such excipient, diluent, carrier, and / or adjuvant.

[0095] As used herein, the terms "extended release," "delayed release," and "controlled release" refer to an extended or sustained release of a therapeutic agent or API of a pharmaceutical formulation. These terms may further refer to compositions that provide an extended or prolonged duration of action, such as the pharmacokinetic (PK) parameters of a pharmaceutical composition comprising a therapeutically effective amount of an active pharmaceutical ingredient as described herein.

[0096] In general, a reference or description of a particular element, such as hydrogen or H, is intended to include all isotopes of that element. For example, if an R group is defined to include hydrogen or H, it also includes deuterium and tritium. Thus, tritium, 14 C. 32 P and 35 Compounds containing radioisotopes such as S are within the scope of the present technology. Procedures for incorporating such labels into the compounds of the present technology will be readily apparent to those of skill in the art based on the disclosure herein.

[0097] Unless specific stereochemistry is explicitly stated, all chiral, diastereomeric and racemic forms of compound are intended.Therefore, the compounds described herein include the optical isomers enriched or resolved at any or all asymmetric atoms as is clear from the description.The racemic mixture of R-enantiomer and S-enantiomer, as well as the enantioenriched stereoisomeric mixture containing R- and S-enantiomer, as well as individual optical isomers can be isolated or synthesized so as to be substantially free of their enantiomeric or diastereomeric partners, and all of these stereoisomers are within the scope of the present technology.

[0098] The compounds described herein may exist as solvates, particularly hydrates, and all such solvates and hydrates are intended unless otherwise specified.Hydrates may form during the preparation of the compounds or compositions containing the compounds, or hydrates may form over time due to the hygroscopic nature of the compounds.The compounds of the present technology may also exist as organic solvent solvates, including DMF, ether, and alcohol solvates, among others.The identification and preparation of any particular solvate is within the skill of a person skilled in the art of synthetic organic or medicinal chemistry.

[0099] As described herein, this text relates to various embodiments of the compounds, compositions and methods.The various embodiments described are meant to provide various examples and should not be interpreted as descriptions of alternative species.Rather, it should be noted that the descriptions of the various embodiments provided herein may be of overlapping scope.The embodiments discussed herein are merely illustrative and are not meant to limit the scope of the present technology.

[0100] 4.2. Summary of experimental observations We demonstrate that TQS-168 induces PGC-1α gene and protein expression in vitro in a murine myeloid cell line, BV2, at concentrations ranging from 0.7 μM to 20 μM, and that TQS-168 inhibits LPS-induced secretion of proinflammatory cytokines from BV2 cells and human primary myeloid cells in vitro at concentrations ranging from 0.3 μM to 20 μM.

[0101] These in vitro experiments predict that plasma and brain concentrations of TQS-168 in the range of 0.3–20 μM should suppress bone marrow-mediated neuroinflammation.

[0102] When administered orally at 25-50 mg / kg, TQS-168 has previously been shown to suppress bone marrow-mediated inflammation and reduce disease severity in mouse models of neurodegenerative diseases including Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis (ALS). U.S. Patent No. 10,272,070. The inventors have measured plasma, liver, and brain concentrations of TQS-168 following administration of a single oral dose of 25 mg / kg to mice, a dose that has previously been found to provide a therapeutic effect. At this previously established effective dose, the mean plasma C of TQS-168 was 1.2-fold higher than that of TQS-168 in mice. max was 93.4ng / ml, or 0.37μM, and the brain C max The higher concentration, 542.0 ng / ml, or 2.16 μM, has been shown to induce PGC-1α protein expression in vitro and to reduce LPS-mediated inflammatory cytokine release within that concentration range. These in vivo data confirm that plasma concentrations of TQS-168 in the range of 0.3 μM to 20 μM should suppress bone marrow-mediated neuroinflammation. Evidence of preferential accumulation in the brain suggests that plasma concentrations of TQS-168 below 0.37 μM may also be effective in treating neuroinflammation.

[0103] We also demonstrate that TQS-621, an early phase 1 metabolite of TQS-168, potently inhibits LPS-induced IL-6 and TNFα secretion from primary human PBMCs. These data demonstrate that at least a portion of the therapeutic effects observed following oral administration of TQS-168 may be attributable to the activity of the metabolite TQS-621.

[0104] The data show that oral administration of TQS-168 in solution results in higher TQS-168 and TQS-621C efficacy than that seen with two different suspension formulations. max and total drug exposure.

[0105] The inventors have also conducted a Phase 1 human clinical trial measuring plasma concentrations of TQS-168 and the active metabolite TQS-621 in three different formulations to demonstrate that pharmacodynamically relevant plasma concentrations can be achieved in oral suspensions of several solid formulations of the API.

[0106] 4.3. Methods of Reducing Neuroinflammation and / or Treating Neurodegenerative Diseases 4.3.1. Pharmacokinetic Dosage Regimen Thus, in a first aspect, there is provided a method for reducing neuroinflammation and / or treating a neurodegenerative disease in a subject, the method comprising administering to a subject having a neuroinflammation and / or neurodegenerative disease a compound of formula (I) [ka] (TQS-168) (MW250.3), or a pharma- ceutical acceptable salt thereof, is administered to a subject in need thereof, the subject being evaluated for the following: (a) a mean peak blood or plasma TQS-168 concentration (C max In certain embodiments, the amount is: (a) an average peak blood or plasma TQS-168 concentration (C) of at least 50 nM (12.515 ng / mL). max ) and (b) the C of TQS-168 in blood or plasma for 360 min or less. max Average time to max In some embodiments, C max and T max is measured in plasma.

[0107] TQS-168 contains 100nM(25.03ng / mL) and 150nM(37). 545ng / mL)、200nM(50.06ng / mL)、250nM(62,575ng / mL)、300nM(75.09ng / mL). L)、350nM(87,605ng / mL)、400nM(100.12ng / mL)、450nM(112,635ng / mL)、5 00nM(125.15ng / mL)、550nM(137.665ng / mL)、600nM(150.18ng / mL)、650nM (162,695ng / mL)、700nM(175.21ng / mL)、750nM(187,725ng / mL)、800nM(2 00.24ng / mL, 850nM(212,755ng / mL), 900nM(225.27ng / mL), 950nM(237.7). 85ng / mL)、1µM(250.3ng / mL)、2µM(500.6ng / mL)、2.5µM(625.75ng / mL)、3µ M(750.9ng / mL), 3.5µM(876.05ng / mL), 4µM(1001.2ng / mL), 4.5µM(1126.3). 5ng / mL)、5µM(1151.5ng / mL)、5.5µM(1376.65ng / mL)、6µM(1501.8ng / mL) 6.5µM(1626.95ng / mL) 7µM(1752.1ng / mL) 7.5µM(1877.25ng / mL) 8µM( 2002.4ng / mL)、8.5µM(2127.55ng / mL)、9µM(2252.7ng / mL)、9.5µM(2377.8). 5ng / mL)、10µM(2503ng / mL)、10.5µM(2628.15ng / mL)、11µM(2753.3ng / mL) 11.5µM(2878.45ng / mL) 12µM(3003.6ng / mL) 12.5µM(3128.75ng / mL) 13µM(3253.9ng / mL)、13.5µM(3379.05ng / mL)、14µM(3504.2ng / mL)、14.5µ M(3629.35ng / mL)、15µM(3754.5ng / mL)、15.5µM(3879.65ng / mL)、16µM(40 04.8ng / mL)、16.5µM(4129.95ng / mL)、17µM(4255.1ng / mL)、17.5µM(4380.25ng / mL), 18μM (4505.4ng / mL), 18.5μM (4630.55ng / mL), 19μM (4755.7ng / mL), 19.5μM (4880.85 ng / mL), 20μM (5006ng / mL), 20.5μM (5131.15ng / mL), 21μM (5256.3ng / mL), 21.5μM (5381.45ng / mL L), 22 μM (5506.6 ng / mL), 22.5 μM (5631.75 ng / mL), 23 μM (5756.9 ng / mL), 23.5 μM (5882.05 ng / mL), 24 μM (6007.2 ng / mL), 24.5 μM (6132.35 ng / mL), or 25 μM (6275.5 ng / mL) of TQS-168 in mean blood or plasma C. max The compound is administered in an amount that results in

[0108] In certain embodiments, TQS-168 or a salt thereof is administered in an amount that results in a mean blood or plasma Cmax of TQS-168 of at least 3.5 μM (876.05 ng / mL), 4 μM (1001.2 ng / mL), 4.5 μM (1126.35 ng / mL), 5 μM (1151.5 ng / mL), 5.5 μM (1376.65 ng / mL), 6 μM (1501.8 ng / mL), 6.5 μM (1626.95 ng / mL), 7 μM (1752.1 ng / mL), 7.5 μM (18778.25 ng / mL), or 8 μM (2002.4 ng / mL) following administration.

[0109] In specific embodiments, TQS-168 or a salt thereof is administered in an amount that results in a mean blood or plasma Cmax of TQS-168 of at least 4 μM (1001.2 ng / mL), 4.5 μM (1126.35 ng / mL), 5 μM (1151.5 ng / mL), or 5.5 μM (1376.65 ng / mL) following administration.

[0110] In some embodiments, TQS-168 or a salt thereof is administered in an amount that results in a mean blood or plasma Cmax of TQS-168 of between 2 μM (500.6 ng / mL) and 8 μM (2002.4 ng / mL), between 2.5 μM (625.75 ng / mL) and 7.5 μM (1877.25 ng / mL), between 3 μM (750.9 ng / mL) and 7 μM (1752.1 ng / mL), 3.5 μM (876.05 ng / mL), 6.5 μM (1626.95 ng / mL), or 4 μM (1001.2 ng / mL) and 6 μM (1501.8 ng / mL) following administration. In a specific embodiment, TQS-168 or a salt thereof is administered in an amount that results in a mean blood or plasma Cmax of TQS-168 of between 4 μM (1001.2 ng / mL) and 5 μM (1151.5 ng / mL) following administration.

[0111] In certain embodiments, TQS-168 or a salt thereof is administered in an amount that results in a mean blood or plasma Cmax of TQS-168 of about 4.5 μM (1126.35 ng / mL) following administration.

[0112] In some embodiments, TQS-168 or a salt thereof is administered in an amount that results in a mean blood or plasma Cmax of TQS-168 of at least 700ng / mL, 750ng / mL, 800ng / mL, 850ng / mL, 900ng / mL, 950ng / mL, 1000ng / mL, 1500ng / mL, or 2000ng / mL after administration. In certain embodiments, TQS-168 or a salt thereof is administered in an amount that results in a mean blood or plasma Cmax of TQS-168 of at least 900ng / mL, 950ng / mL, 1000ng / mL, 1500ng / mL, or 2000ng / mL after administration. In certain embodiments, TQS-168 or a salt thereof is administered in an amount that results in a mean blood or plasma Cmax of TQS-168 of at least 1000ng / mL, 1100ng / mL, 1200ng / mL, 1300ng / mL, 1400ng / mL, 1500ng / mL, 1600ng / mL, 1700ng / mL, 1800ng / mL, 1900ng / mL, or 2000ng / mL after administration. In certain embodiments, TQS-168 or a salt thereof is administered in an amount that results in a mean blood or plasma Cmax of TQS-168 of between 900ng / mL and 1300ng / mL or between 1000ng / mL and 1200ng / mL after administration.

[0113] In some embodiments, TQS-168 or a salt thereof is administered in an amount that results in a mean blood or plasma Cmax of TQS-168 after administration of about 1100 ng / mL.

[0114] In some embodiments, TQS-168 or a salt thereof is administered at a concentration of at least 50 nM, 100 nM, 150 nM, 200 nM, 250 nM, 300 nM, 350 nM, 400 nM, 450 nM, 500 nM, 550 nM, 600 nM, 650 nM, 700 nM, 750 nM, 800 nM, 850 nM, 900 nM, 950 nM, 1 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5μM, 9μM, 9.5μM, 10μM, 10.5μM, 11μM, 11.5μM, 2μM, 12.5μM, 13μM, 13.5μM, 14μM, 14.5μM, 15μM, 15.5μM, 16μM, 16.5μM, 17μM, 17.5 Average brain C of TQS-168 at μM, 18 μM, 18.5 μM, 19 μM, 19.5 μM, 20 μM, 20.5 μM, 21 μM, 21.5 μM, 22 μM, 22.5 μM, 23 μM, 23.5 μM, 24 μM, 24.5 μM, or 25 μM max The compound is administered in an amount that results in

[0115] In some embodiments, TQS-168 or a salt thereof is administered in an amount that results in a brain-to-plasma ratio of TQS-168 of 0.5 to 10 after administration. In certain embodiments, TQS-168 is administered in an amount that results in a brain-to-plasma ratio of TQS-168 of 0.5, 0.75, 1.0, 1.25, 1.5, 1.75, 2.0, 2.25, 2.5, 2.75, 3.0, 3.25, 3.5, 3.75, 4.0, 4.25, 4.5, 4.75, or 5 after administration. In certain embodiments, TQS-168 is administered in an amount that results in a brain-to-plasma ratio of TQS-168 of at least 1.0, at least 1.5, at least 2.0, at least 2.5, at least 3.0, at least 3.5, at least 4.0, at least 4.5, or at least 5.0 after administration.

[0116] In some embodiments, after administration of TQS-168 or a salt thereof, an AUC of TQS-168 measured in plasma of at least 2000ng·hr / ml, 2500ng·hr / ml, 3000ng·hr / ml, 3500ng·hr / ml, 4000ng·hr / ml, 4500ng·hr / ml, 5000ng·hr / ml, 5500ng·hr / ml, 6000ng·hr / ml, 6500ng·hr / ml, 7000ng·hr / ml, 7500ng·hr / ml, 8000ng·hr / ml, 8500ng·hr / ml, or 9000ng·hr / ml. 0-t In certain embodiments, the TQS-168 or a salt thereof is administered in an amount that results in an AUC of TQS-168 in plasma of at least 4000 ng·hr / ml, 4500 ng·hr / ml, 5000 ng·hr / ml, 5500 ng·hr / ml, 6000 ng·hr / ml, 6500 ng·hr / ml, or 7000 ng·hr / ml after administration. 0-t The compound is administered in an amount that results in

[0117] In certain embodiments, TQS-168 or a salt thereof is administered to a subject in need thereof, the subject being administered an AUC of TQS-168 in plasma of between 4000 ng·hr / ml and 8000 ng·hr / ml after administration. 0-t In certain embodiments, the TQS-168 or salt thereof is administered in an amount that results in an AUC of TQS-168 in plasma of between 5000 ng·hr / ml and 7000 ng·hr / ml after administration. 0-t In a specific embodiment, the TQS-168 or salt thereof is administered in an amount that results in an AUC of TQS-168 in plasma of about 6000 ng·hr / ml after administration. 0-t The compound is administered in an amount that results in

[0118] In some embodiments, TQS-168 or a salt thereof is administered to a subject in need of treatment with an immunization assay to determine the mean T of TQS-168 in blood or plasma for up to 360 minutes after administration. maxIn certain embodiments, the TQS-168 or salt thereof is administered in an amount that results in a mean T in blood or plasma of less than or equal to 360 minutes, 350 minutes, 340 minutes, 330 minutes, 320 minutes, 310 minutes, 300 minutes, 290 minutes, 280 minutes, 270 minutes, 260 minutes, 250 minutes, 225 minutes, 200 minutes, or 180 minutes. max In certain embodiments, the TQS-168 or a salt thereof is administered in a formulation that results in a mean T in blood or plasma of 90 minutes, 60 minutes, or 45 minutes or less. max The compound is administered in a formulation which produces

[0119] In certain embodiments, TQS-168 or a salt thereof is administered to a subject at or below 120 minutes, 90 minutes, or 60 minutes after administration to determine the mean T of TQS-168 in blood or plasma. max In a specific embodiment, the TQS-168 or a salt thereof is administered in an amount that results in a mean T of about 60 minutes after administration. max The compound is administered in an amount that results in

[0120] In another embodiment, a method is provided for treating neuroinflammation and / or treating a neurodegenerative disease in a subject, comprising administering to a subject having a neuroinflammation and / or neurodegenerative disease a pharmaceutical composition comprising TQS-168, or a pharma- ceutical acceptable salt thereof, followed by: (a) administering at least 50 nM of a compound of formula (II) to the subject; [ka] The mean peak plasma concentration (C max ) and (b) the C max Average time to max ) in an amount that provides

[0121] In some embodiments, TQS-168 or a salt thereof is administered at a concentration of at least 100 nM, 150 nM, 200 nM, 250 nM, 300 nM, 350 nM, 400 nM, 450 nM, 500 nM, 550 nM, 600 nM, 650 nM, 700 nM, 750 nM, 800 nM, 850 nM, 900 nM, 950 nM, 1 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, Mean blood or plasma C of TQS-621 at 9 μM, 9.5 μM, 10 μM, 10.5 μM, 11 μM, 11.5 μM, 12 μM, 12.5 μM, 13 μM, 13.5 μM, 14 μM, 14.5 μM, 15 μM, 15.5 μM, 16 μM, 16.5 μM, 17 μM, 17.5 μM, 18 μM, 18.5 μM, 19 μM, 19.5 μM, 20 μM, 20.5 μM, 21 μM, 21.5 μM, 22 μM, 22.5 μM, 23 μM, 23.5 μM, 24 μM, 24.5 μM, or 25 μM max The compound is administered in an amount that results in

[0122] In some embodiments, TQS-168 or a salt thereof is administered to a subject with a mean blood or plasma C of at least 75 ng / mL, 100 ng / mL, 125 ng / mL, 150 ng / mL, 175 ng / mL, 200 ng / mL, 225 ng / mL, 250 ng / mL, 250 ng / mL, 300 ng / mL, 350 ng / mL, 400 ng / mL, 450 ng / mL, 500 ng / mL, 550 ng / mL, or 600 ng / mL of TQS-621 after administration. max The compound is administered in an amount that results in

[0123] In some embodiments, TQS-168 or a salt thereof is administered to a subject with a mean blood or plasma C of 100-700 ng / mL, 200-600 ng / mL, or 300-500 ng / mL of TQS-621 after administration. max The compound is administered in an amount that results in

[0124] In some embodiments, TQS-168 or a salt thereof is administered at least 50 nM, 100 nM, 150 nM, 200 nM, 250 nM, 300 nM, 350 nM, 400 nM, 450 nM, 500 nM, 550 nM, 600 nM, 650 nM, 700 nM, 750 nM, 800 nM, 850 nM, 900 nM, 950 nM, 1 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 21 μM, 22 μM, 23 μM, 24 μM, 25 μM, 26 μM, 27 μM, 28 μM, 29 μM, 30 μM, 31 μM, 32 μM, 33 μM, 34 μM, 35 μM, 36 μM, 37 μM, 38 μM, 39 μM, 40 μM, 41 μM, 42 μM, 43 μM, 44 μM, 45 μM, 46 μM, 47 μM, 48 μM, 49 μM, 50 μM, 51 μM, 52 μM, 53 μM, 54 μ .5μM, 9μM, 9.5μM, 10μM, 10.5μM, 11μM, 11.5μM, 12μM, 12.5μM, 13μM, 13.5μM, 14μM, 14.5μM, 15μM, 15.5μM, 16μM, 16.5μM, 17μM, 17.5 Average brain C of TQS-621 at μM, 18 μM, 18.5 μM, 19 μM, 19.5 μM, 20 μM, 20.5 μM, 21 μM, 21.5 μM, 22 μM, 22.5 μM, 23 μM, 23.5 μM, 24 μM, 24.5 μM, or 25 μM max The compound is administered in an amount that results in

[0125] In some embodiments, TQS-168 or a salt thereof is administered in an amount that results in a brain-to-plasma ratio of TQS-621 after administration of 0.5 to 10. In certain embodiments, TQS-168 is administered in an amount that results in a brain-to-plasma ratio of TQS-168 after administration of 0.5, 0.75, 1.0, 1.25, 1.5, 1.75, 2.0, 2.25, 2.5, 2.75, 3.0, 3.25, 3.5, 3.75, 4.0, 4.25, 4.5, 4.75, or 5. In certain embodiments, TQS-168 is administered in an amount that results in a brain-to-plasma ratio of TQS-168 after administration of at least 1.0, at least 1.5, at least 2.0, at least 2.5, at least 3.0, at least 3.5, at least 4.0, at least 4.5, or at least 5.0.

[0126] In some embodiments, TQS-168 or a salt thereof is administered to a subject in need of treatment with an immunization assay to determine the mean T of TQS-621 in blood or plasma for up to 360 minutes after administration. maxIn certain embodiments, the TQS-168 or salt thereof is administered in an amount that results in a mean T in blood or plasma of less than or equal to 360 minutes, 350 minutes, 340 minutes, 330 minutes, 320 minutes, 310 minutes, 300 minutes, 290 minutes, 280 minutes, 270 minutes, 260 minutes, 250 minutes, 225 minutes, 200 minutes, or 180 minutes. max In certain embodiments, the TQS-168 or a salt thereof is administered in a formulation that results in a mean T in blood or plasma of 90 minutes, 60 minutes, or 45 minutes or less. max The compound is administered in a formulation which produces

[0127] In another embodiment, a method of treating a neuroinflammatory and / or neurodegenerative disease in a subject is provided, the method comprising administering to a subject having a neuroinflammatory and / or neurodegenerative disease a pharmaceutical composition comprising TQS-168 or a pharma- ceutical acceptable salt thereof, (a) achieving a mean peak plasma concentration (C max ) and (b) the C max Average time to max ) with (c) a mean peak concentration (C max ) and (d) the C max Average time to max ) in an amount that provides

[0128] In some embodiments, TQS-168 or a salt thereof is administered at a concentration of at least 100 nM, 150 nM, 200 nM, 250 nM, 300 nM, 350 nM, 400 nM, 450 nM, 500 nM, 550 nM, 600 nM, 650 nM, 700 nM, 750 nM, 800 nM, 850 nM, 900 nM, 950 nM, 1 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, Mean blood or plasma C of TQS-168 at 9 μM, 9.5 μM, 10 μM, 10.5 μM, 11 μM, 11.5 μM, 12 μM, 12.5 μM, 13 μM, 13.5 μM, 14 μM, 14.5 μM, 15 μM, 15.5 μM, 16 μM, 16.5 μM, 17 μM, 17.5 μM, 18 μM, 18.5 μM, 19 μM, 19.5 μM, 20 μM, 20.5 μM, 21 μM, 21.5 μM, 22 μM, 22.5 μM, 23 μM, 23.5 μM, 24 μM, 24.5 μM, or 25 μM max The compound is administered in an amount that results in

[0129] In some embodiments, TQS-168 or a salt thereof is administered at least 50 nM, 100 nM, 150 nM, 200 nM, 250 nM, 300 nM, 350 nM, 400 nM, 450 nM, 500 nM, 550 nM, 600 nM, 650 nM, 700 nM, 750 nM, 800 nM, 850 nM, 900 nM, 950 nM, 1 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 21 μM, 22 μM, 23 μM, 24 μM, 25 μM, 26 μM, 27 μM, 28 μM, 29 μM, 30 μM, 31 μM, 32 μM, 33 μM, 34 μM, 35 μM, 36 μM, 37 μM, 38 μM, 39 μM, 40 μM, 41 μM, 42 μM, 43 μM, 44 μM, 45 μM, 46 μM, 47 μM, 48 μM, 49 μM, 50 μM, 51 μM, 52 μM, 53 μM, 54 μ .5μM, 9μM, 9.5μM, 10μM, 10.5μM, 11μM, 11.5μM, 12μM, 12.5μM, 13μM, 13.5μM, 14μM, 14.5μM, 15μM, 15.5μM, 16μM, 16.5μM, 17μM, 17.5 Average brain C of TQS-168 at μM, 18 μM, 18.5 μM, 19 μM, 19.5 μM, 20 μM, 20.5 μM, 21 μM, 21.5 μM, 22 μM, 22.5 μM, 23 μM, 23.5 μM, 24 μM, 24.5 μM, or 25 μM max The compound is administered in an amount that results in

[0130] In some embodiments, TQS-168 or a salt thereof is administered in an amount that results in a brain-to-plasma ratio of TQS-168 of 0.5 to 10 after administration. In certain embodiments, TQS-168 is administered in an amount that results in a brain-to-plasma ratio of TQS-168 of 0.5, 0.75, 1.0, 1.25, 1.5, 1.75, 2.0, 2.25, 2.5, 2.75, 3.0, 3.25, 3.5, 3.75, 4.0, 4.25, 4.5, 4.75, or 5 after administration. In certain embodiments, TQS-168 is administered in an amount that results in a brain-to-plasma ratio of TQS-168 of at least 1.0, at least 1.5, at least 2.0, at least 2.5, at least 3.0, at least 3.5, at least 4.0, at least 4.5, or at least 5.0 after administration.

[0131] In some embodiments, TQS-168 or a salt thereof is administered at a concentration of at least 100 nM, 150 nM, 200 nM, 250 nM, 300 nM, 350 nM, 400 nM, 450 nM, 500 nM, 550 nM, 600 nM, 650 nM, 700 nM, 750 nM, 800 nM, 850 nM, 900 nM, 950 nM, 1 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, Mean blood or plasma C of TQS-621 at 9 μM, 9.5 μM, 10 μM, 10.5 μM, 11 μM, 11.5 μM, 12 μM, 12.5 μM, 13 μM, 13.5 μM, 14 μM, 14.5 μM, 15 μM, 15.5 μM, 16 μM, 16.5 μM, 17 μM, 17.5 μM, 18 μM, 18.5 μM, 19 μM, 19.5 μM, 20 μM, 20.5 μM, 21 μM, 21.5 μM, 22 μM, 22.5 μM, 23 μM, 23.5 μM, 24 μM, 24.5 μM, or 25 μM max The compound is administered in an amount that results in

[0132] In some embodiments, TQS-168 or a salt thereof is administered at least 50 nM, 100 nM, 150 nM, 200 nM, 250 nM, 300 nM, 350 nM, 400 nM, 450 nM, 500 nM, 550 nM, 600 nM, 650 nM, 700 nM, 750 nM, 800 nM, 850 nM, 900 nM, 950 nM, 1 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 21 μM, 22 μM, 23 μM, 24 μM, 25 μM, 26 μM, 27 μM, 28 μM, 29 μM, 30 μM, 31 μM, 32 μM, 33 μM, 34 μM, 35 μM, 36 μM, 37 μM, 38 μM, 39 μM, 40 μM, 41 μM, 42 μM, 43 μM, 44 μM, 45 μM, 46 μM, 47 μM, 48 μM, 49 μM, 50 μM, 51 μM, 52 μM, 53 μM, 54 μ .5μM, 9μM, 9.5μM, 10μM, 10.5μM, 11μM, 11.5μM, 12μM, 12.5μM, 13μM, 13.5μM, 14μM, 14.5μM, 15μM, 15.5μM, 16μM, 16.5μM, 17μM, 17.5 Average brain C of TQS-621 at μM, 18 μM, 18.5 μM, 19 μM, 19.5 μM, 20 μM, 20.5 μM, 21 μM, 21.5 μM, 22 μM, 22.5 μM, 23 μM, 23.5 μM, 24 μM, 24.5 μM, or 25 μM max The compound is administered in an amount that results in

[0133] In some embodiments, TQS-168 or a salt thereof is administered in an amount that results in a brain-to-plasma ratio of TQS-621 after administration of 0.5 to 10. In certain embodiments, TQS-168 is administered in an amount that results in a brain-to-plasma ratio of TQS-168 after administration of 0.5, 0.75, 1.0, 1.25, 1.5, 1.75, 2.0, 2.25, 2.5, 2.75, 3.0, 3.25, 3.5, 3.75, 4.0, 4.25, 4.5, 4.75, or 5. In certain embodiments, TQS-168 is administered in an amount that results in a brain-to-plasma ratio of TQS-168 after administration of at least 1.0, at least 1.5, at least 2.0, at least 2.5, at least 3.0, at least 3.5, at least 4.0, at least 4.5, or at least 5.0.

[0134] Inhibition of inflammation in the periphery can be beneficial in the treatment of neuroinflammation.For example, fingolimod, which is currently approved for treating relapsing-remitting multiple sclerosis (MS), acts to reduce the lymphocyte escape from lymph nodes and thereby alleviate MS pathology.Therefore, in some embodiments, TQS-168 is administered in an amount that produces optimal concentrations of TQS-168 and metabolite TQS-621 in both peripheral and central compartments. [Table 1-1] In various embodiments, TQS-168 is Plasma TQS-168: Brain TQS-168 Plasma TQS-621: Brain TQS-621 Plasma TQS-168: Plasma TQS-621 TQS-168 in the brain: TQS-621 in the brain The compound is administered in an amount that provides an optimal concentration ratio of one or more of the following:

[0135] Oral Dosage In various embodiments, the daily oral dose of TQS-168 is at least 0.5 mg / kg. In various embodiments, the oral dose of TQS-168 is at least 1 mg / kg. In certain embodiments, the dose is at least 2 mg / kg, at least 3 mg / kg, at least 4 mg / kg, at least 5 mg / kg, at least 6 mg / kg, at least 7 mg / kg, at least 8 mg / kg, at least 9 mg / kg, or at least 10 mg / kg.

[0136] In various embodiments, the daily oral dose of TQS-168 is at least 10 mg / kg. In certain embodiments, the dose is at least 15 mg / kg, at least 20 mg / kg, at least 25 mg / kg, 30 mg / kg, at least 35 mg / kg, at least 40 mg / kg, at least 45 mg / kg, at least 50 mg / kg, at least 55 mg / kg, at least 60 mg / kg, at least 65 mg / kg, at least 70 mg / kg, at least 75 mg / kg, at least 80 mg / kg, at least 85 mg / kg, at least 90 mg / kg, at least 95 mg / kg, at least 100 mg / kg, at least 150 mg / kg, at least 175 mg / kg, or at least 200 mg / kg. In certain embodiments, the dose is 250 mg / kg, 300 mg / kg, 350 mg / kg, 400 mg / kg, 450 mg / kg, 500 mg / kg, 600 mg / kg, 650 mg / kg, 700 mg / kg, 750 mg / kg, 800 mg / kg, 850 mg / kg, 900 mg / kg, 950 mg / kg, or 1000 mg / kg. In certain embodiments, the oral dose is 0.5 mg / kg to 100 mg / kg per day. In certain embodiments, the oral dose is 2 mg / kg to 100 mg / kg per day. In certain embodiments, the oral dose is 25 mg / kg to 1000 mg / kg per day.

[0137] In various embodiments, the oral daily dose of TQS-168 is 25 mg / kg. In certain embodiments, the dose is at least 25 mg / kg. In certain embodiments, the dose is at least 50 mg / kg, at least 100 mg / kg, at least 150 mg / kg, at least 175 mg / kg, or at least 200 mg / kg. In certain embodiments, the dose is 250 mg / kg, 500 mg / kg, 750 mg / kg, or 1000 mg / kg. In certain embodiments, the oral dose is between 25 mg / kg and 1,000 mg / kg per day.

[0138] In various embodiments, the oral daily dose is 10-5000mg. In certain embodiments, the dose is 10mg, 15mg, 20mg, 25mg, 50mg, 75mg, 100mg, 125mg, 150mg, 175mg, 200mg, 225mg, 250mg, 275mg, 300mg, 325mg, 350mg, 375mg, 400mg, 450mg, 500mg, 550mg, 600mg, 650mg, 700mg, 750mg, 800mg, 850mg, 900mg, 950mg, or 1000mg. In certain embodiments, the dose is 1500mg, 2000mg, 2500mg, 3000mg, 3500mg, 4000mg, 4500mg, or 5000mg.

[0139] In various embodiments, the daily dose is between 25 and 2000 mg. In certain embodiments, the dose is 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 350 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 825 mg, 900 mg, 925 mg, 950 mg, 975 mg, or 1000 mg.

[0140] In certain embodiments, the oral daily dose is 200-800mg. In certain embodiments, the oral daily dose is 350mg, 360mg, 370mg, 380mg, 390mg, 400mg, 410mg, 420mg, 430mg, 440mg, 450mg, 460mg, 470mg, 480mg, 490mg, or 500mg. In certain embodiments, the dose is 400mg or 450mg. In certain embodiments, the oral daily dose is 400mg or 450mg for spray-dried dispersion formulations.

[0141] 4.3.3. Oral dosage forms In some embodiments, TQS-168 or a salt thereof is administered in a suspension. In other embodiments, TQS-168 or a salt thereof is administered in a solution. In some embodiments, TQS-168 or a salt thereof is administered in a solid dosage form. In certain embodiments, the solid dosage form is a capsule. In certain embodiments, the solid dosage form is a tablet. In specific embodiments, TQS-168 is in crystalline or amorphous form. In certain embodiments, TQS-168 is in amorphous form.

[0142] 4.3.4. Patient In various embodiments, the subject has neuroinflammation.In certain embodiments, the subject does not have a diagnosed neurodegenerative disease.In certain embodiments, the subject does not have a diagnosed neurodegenerative disease and is at least 40, 45, 50, 55, 60, 65, 70, or 75 years old.In certain embodiments, the subject does not have a diagnosed neurodegenerative disease but has one or more signs or symptoms of cognitive impairment.In a specific embodiment, the subject has mild cognitive impairment (MCI).

[0143] In various embodiments, the subject has a neurodegenerative disease. In certain embodiments, the neurodegenerative disease is selected from motor neuron disease, amyotrophic lateral sclerosis (ALS), Alzheimer's disease, vascular dementia, frontotemporal degeneration (frontotemporal dementia), dementia with Lewy bodies, Parkinson's disease, Huntington's disease, demyelinating disease, and multiple sclerosis (MS).

[0144] In certain embodiments, the subject has a motor neuron disease. In specific embodiments, the subject has ALS. In certain embodiments, the subject has Alzheimer's disease. In certain embodiments, the subject has vascular dementia. In certain embodiments, the subject has frontotemporal dementia (FTD). In certain embodiments, the subject has Lewy body dementia (Lewy body disease). In certain embodiments, the subject has Parkinson's disease. In certain embodiments, the subject has Huntington's disease. In yet additional embodiments, the subject has a demyelinating disease. In one embodiment, the subject has MS.

[0145] 4.3.5. Further embodiments Further embodiments are provided in the following numbered clauses: 1. A method of treating a neuroinflammatory and / or neurodegenerative disease in a subject, comprising: A subject having a neuroinflammatory and / or neurogenerative disorder is administered a compound of formula (I) [ka] (TQS-168) or a pharma- ceutical composition comprising such a pharma- ceutical composition, (a) Mean peak plasma TQS-168 concentration (C) of at least 50 nM max )of, (b) Mean time to Cmax of TQS-168 in plasma (T max ) together with orally administering to the subject a therapeutically effective amount of 2. TQS-168 or its salts are administered (a) a mean plasma TQS-168C of at least 100 nM, 250 nM, or 500 nM max The method of claim 1, wherein the compound is administered in an amount that results in 3. TQS-168 or its salts are administered (a) a mean plasma TQS-168C of at least 750 nM max The method of claim 2, wherein the administering is in an amount that results in 4. TQS-168 or its salts are administered (a) Mean plasma TQS-168C of at least 1 μM max The method of claim 3, wherein the compound is administered in an amount that results in 5. TQS-168 or its salts are administered (a) Mean plasma TQS-168C of at least 5 μM max The method of claim 4, wherein the compound is administered in an amount that results in 6. After administration of TQS-168 or its salt, (a) a mean plasma TQS-168C of at least 7.5 μMmax The method of claim 5, wherein the compound is administered in an amount that results in 7. TQS-168 or its salts are administered (a) Mean plasma TQS-168C of at least 10 μM max The method of claim 6, wherein the compound is administered in an amount that results in 8. TQS-168 or a salt thereof, (b) Mean plasma TQS-168T within 275 min max 8. The method according to any one of clauses 1 to 7, wherein the compound is administered in a formulation which results in 9. TQS-168 or a salt thereof (b) Mean plasma TQS-168T within 250 min max The method according to clause 8, wherein the compound is administered in a formulation which results in 10. TQS-168 or a salt thereof, (b) Mean plasma TQS-168T within 225 min max 10. The method of claim 9, wherein the compound is administered in a formulation which results in 11. TQS-168 or a salt thereof, (b) Mean plasma TQS-168T at 180, 90, 60, or 45 min max 11. The method of claim 10, wherein the compound is administered in a formulation which results in 12. A method of treating a neuroinflammatory and / or neurodegenerative disease in a subject, comprising: A subject having a neuroinflammatory and / or neurogenerative disorder is administered a compound of formula (I) [ka] (TQS-168), or a pharma- ceutical composition comprising ... (a) at least 50 nM of a compound of formula (II) [ka] The mean peak plasma concentration (C max)of, (b) C of TQS-621 in plasma at 360 min or less max Average time to max ) together with orally administering to the subject a therapeutically effective amount of 13. TQS-168 or its salts are administered (a) a mean plasma TQS-621C of at least 100 nM, 250 nM, or 500 nM max 13. The method of claim 12, wherein the compound is administered in an amount that results in 14. TQS-168 or its salts are administered (a) a mean plasma TQS-621C concentration of at least 750 nM max 14. The method of claim 13, wherein the compound is administered in an amount that results in 15. TQS-168 or its salts are administered (a) Mean plasma TQS-621C of at least 1 μM max 15. The method of claim 14, wherein the compound is administered in an amount that results in 16. TQS-168 or its salts are administered (a) Mean plasma TQS-621C of at least 5 μM max 16. The method of claim 15, wherein the compound is administered in an amount that results in 17. TQS-168 or its salts are administered (a) a mean plasma TQS-621C concentration of at least 7.5 μM max 17. The method of claim 16, wherein the compound is administered in an amount that results in 18. TQS-168 or its salts are administered (a) Mean plasma TQS-621C of at least 10 μM max 18. The method of claim 17, wherein the compound is administered in an amount that results in 19. TQS-168 or a salt thereof, (b) Mean plasma TQS-621T within 275 min max 19. The method according to any one of clauses 12 to 18, wherein the composition is administered in a formulation which results in 20. TQS-168 or a salt thereof, (b) Mean plasma T of TQS-168 at 250 min or less max 20. The method of claim 19, wherein the compound is administered in a formulation which results in 21. TQS-168 or a salt thereof, (b) Mean plasma T of TQS-168 at ≤225 min max 21. The method of claim 20, wherein the compound is administered in a formulation which results in 22. TQS-168 or a salt thereof, (b) Mean plasma T of TQS-168 at 180, 90, 60, or 45 min max 22. The method of claim 21, wherein the compound is administered in a formulation which results in 23. A method of treating a neuroinflammatory and / or neurodegenerative disease in a subject, comprising: A subject having a neuroinflammatory and / or neurogenerative disorder is administered a compound of formula (I) [ka] (TQS-168), or a pharma- ceutical composition comprising a medicament for treating a patient with a pulmonary artery disease, After administration (a) Mean peak plasma TQS-168 concentration (C) of at least 50 nM max )of, (b) C of TQS-168 in plasma at 360 min or less max Average time to max ) together with; and (c) at least 50 nM of the compound of formula (II) in plasma [ka] (TQS-621) average peak concentration (C max )of, (d) C of TQS-621 in plasma at 360 min or less max Average time to max ) together with orally administering to the subject a therapeutically effective amount of 24. The method according to any one of clauses 1 to 23, wherein TQS-168 or a salt thereof is administered in a suspension. 25. The method according to any one of clauses 1 to 23, wherein the TQS-168 or a salt thereof is administered in a solution. 26. The method of any one of clauses 1 to 23, wherein TQS-168 or a salt thereof is administered in a solid dosage form. 27. The method according to clause 26, wherein the solid dosage form is a capsule. 28. The method according to clause 26, wherein the solid dosage form is a tablet. 29. The method of any one of clauses 1-24 or 26-28, wherein the TQS-168 is in crystalline or amorphous form. 30. The method of any one of clauses 1-24 or 26-28, wherein the TQS-168 is in amorphous form. 31. The method of any one of clauses 1-30, wherein the subject has a neurodegenerative disease selected from motor neuron disease, amyotrophic lateral sclerosis (ALS), Alzheimer's disease, vascular dementia, frontotemporal degeneration (frontotemporal dementia), dementia with Lewy bodies, Parkinson's disease, Huntington's disease, demyelinating diseases, and multiple sclerosis (MS). 32. The method of clause 31, wherein the subject has a motor neuron disease. 33. The method of clause 31, wherein the subject has ALS. 34. The method of clause 31, wherein the subject has Alzheimer's disease. 35. The method of clause 31, wherein the subject has vascular dementia. 36. The method of clause 31, wherein the subject has frontotemporal dementia (FTD). 37. The method of clause 31, wherein the subject has Lewy body dementia (Lewy body disease). 38. The method of clause 31, wherein the subject has Parkinson's disease. 39. The method of clause 31, wherein the subject has Huntington's disease. 40. The method of clause 31, wherein the subject has a demyelinating disease. 41. The method of clause 31, wherein the subject has MS. 42. The method of any one of clauses 1 to 41, wherein the dose of TQS-168 is at least 0.5 mg / kg. 43. The method of claim 42, wherein the dose of TQS-168 is at least 2 mg / kg. 44. The method of claim 43, wherein the dose of TQS-168 is at least 4 mg / kg. 45. The method of claim 44, wherein the dose of TQS-168 is at least 8 mg / kg. 46. ​​The method according to clause 45, wherein the dose of TQS-168 is at least 12 mg / kg. 47. The method according to clause 46, wherein the dose of TQS-168 is at least 14 mg / kg. 48. The method according to clause 47, wherein the dose of TQS-168 is at least 16 mg / kg. 49. The method according to clause 48, wherein the dose is 2 mg / kg. 50. The method according to clause 49, wherein the dose is 4 mg / kg. 51. The method according to clause 50, wherein the dose is 8 mg / kg. EXAMPLES

[0146] 5. Experimental Example

[0147] Below are examples of specific embodiments for carrying out the present invention. The examples are provided for illustrative purposes only and are not intended to limit the scope of the present invention in any way. Efforts have been made to ensure accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should, of course, be allowed for.

[0148] The practice of the present invention will employ, unless otherwise indicated, conventional methods of protein chemistry, biochemistry, recombinant DNA technology and pharmacology, which are within the skill of the art, and such techniques are fully explained in the literature.

[0149] 5.1. Example 1 TQS-168 at 20 μM induces PGC-1α protein expression in mouse myeloid cell lines in vitro Frozen BV2 mouse microglial cells were thawed and grown in complete medium (RPMI, 10% heat-inactivated FBS, 1% L-glutamine, 1% Pen-Strep) until the proliferation rate reached log phase.

[0150] For protein expression analysis, DMSO (final dilution 1:1000) or 20 μM TQS-168 in DMSO (final dilution 1:1000) was added to cell cultures. After 24 h of stimulation, the supernatant was discarded and cell lysis buffer (Cell Signal) was added to the adherent cells to extract proteins. Total protein was quantified and normalized for all samples by BCA assay.

[0151] Two replicates of untreated cultures and three replicates of TQS-168 treated cultures were examined, with each replicate containing lysates from 5 million BV2 bone marrow cells.

[0152] PGC1α expression was detected by Western blot using anti-PGC-1α antibody (SC13067, Santa Cruz Biotechnology, 1:500 dilution). Anti-β-actin antibody (SC8432, Santa Cruz Biotechnology, 1:2000 dilution) was used to quantify beta-actin, a housekeeping gene whose expression level is not known to be affected by TQS-168. A representative Western blot is shown in FIG. 1.

[0153] As shown, TQS-168 at 20 μM induces PGC-1α protein expression in mouse BV2 microglial cells in vitro.

[0154] 5.2. Example 2 TQS-168 induces PGC-1α protein expression in mouse myeloid cell lines in vitro at concentrations between 0.7 and 20 μM For protein expression analysis, DMSO (final dilution 1:1000) or 20 μM, 6.8 μM, 2.2 μM, and 0.7 μM TQS-168 in DMSO (final dilution 1:1000) were added to BV2 cell cultures. After 24 h of stimulation, the supernatant was discarded and cell lysis buffer (Cell Signal) was added to the adherent cells to extract proteins. Total protein was quantified and normalized for all samples by BCA assay. Subsequently, PGC-1α was detected by Western blot using anti-PGC-1α antibody (SC13067, Santa Cruz Biotechnology, dilution 1:500). β-actin was detected with anti-β-actin antibody (SC8432, Santa Cruz Biotechnology, dilution 1:2000). The results are shown in Figure 2. All procedures were performed with standard molecular biology protocols for protein expression studies.

[0155] Figure 3 is a bar graph quantifying protein expression levels measured from scans of the Western blots shown in Figure 2. PD refers to PD169316, a p38 MAPK inhibitor.

[0156] Figures 2 and 3 demonstrate that TQS-168 induces PGC-1α protein expression in mouse BV2 cells in vitro at concentrations ranging from 0.7 μM to 20 μM.

[0157] 5.3. Example 3 TQS-168 inhibits inflammatory cytokine secretion from LPS-stimulated BV2 cells in vitro (CBA assay) at concentrations ranging from 1 μM to 20 μM. Lipopolysaccharide (LPS) is the natural ligand for the TLR4 / CD14 complex, which is highly expressed on myeloid cells.

[0158] LPS was used to induce cytokine secretion from BV-2 cells. Cells were incubated with various concentrations of TQS-168 to evaluate whether TQS-168 could suppress LPS-promoted release of various cytokines from in vitro cell cultures. Cytokine secretion was measured by cytometric bead array (CBA) fluorescence-activated cell sorting (FACS).

[0159] Briefly, a vial of BV2 frozen stock (1 million cells per ml in complete medium) was thawed into 10 mL of complete medium per vial for a total of 4 vials. The vials were then centrifuged at 1800 rpm for 3 minutes to wash off the freezing medium. The 4 vials were then pooled in 25 mL of complete medium. TQS-168 was prepared in DMSO.

[0160] Cells were incubated for 24 hours in the presence of medium (negative control), LPS (positive control), DMSO+LPS (positive control, control with the presence of DMSO in the TQS-168 stock solution), or LPS+TQS-168 at final concentrations of 1 μM, 5 μM, 10 μM, and 20 μM.

[0161] After incubating the well plates overnight for 24 hours, they were centrifuged at 1800 rpm for 5 minutes. Then, 150 μl from each well was transferred to a new set of plates, of which 50 μl was used for CBA. The plates were then centrifuged with the cells, followed by the addition of 100 μL DAPI (50 ml PBS + 10 μL DAPI stock at 1:5000 dilution). The plates were then incubated in the dark for 5 minutes, followed by the addition of 100 μL PBS and centrifugation at 1800 rpm for 5 minutes. Finally, the plates were resuspended in 200 μL PBS and tested.

[0162] Multiplexed cytometric bead array (CBA) assays were performed by standard techniques to detect the presence in cell culture medium of secreted murine TNFα, IL-6, IFNγ, IL-12, monocyte chemotactic protein 1 (MCP-1), and IL-10.

[0163] conclusion As shown in Figure 4, BV2 cells treated with 100 ng / ml LPS showed a strong release of TNFα and IL-6 proinflammatory cytokines, whereas the secretion of INF-γ, IL-10, IL-12 and MCP-1 was not significantly induced. TQS-168 could reduce the LPS-induced secretion of TNFα and IL-6 from BV2 cells at concentrations ranging from 1 μM to 20 μM. The reduction of both TNFα (Figure 4a) and IL-6 (Figure 4b) by TQS-168 was concentration-dependent.

[0164] 5.4. Example 4 TQS-168 inhibits LPS-induced TNFα secretion from BV2 myeloid cells in vitro at 5 and 20 μM (CBA assay) A total of 4–11 replicates of BV2 cultures in different conditions were investigated. Frozen BV2 microglial cell lines were thawed and grown in complete medium (RPMI, 10% heat-inactivated FBS, 1% L-glutamine, 1% Pen-Strep) until the growth rate reached logarithmic phase.

[0165] For TNFα stimulation, cells were stimulated with LPS 100ng / mL for 24 hours. DMSO (final dilution 1:1000) or TQS-168 in DMSO at 5μM or 20μM (final dilution 1:1000) was added to the cell cultures. After 24 hours of stimulation, supernatants were collected for cytokine analysis with the CBA assay (BD Biosciences) according to the manufacturer's protocol. TNFα expression was normalized to the DMSO treatment condition.

[0166] ANOVA was used for statistical analysis with a significance threshold of p-value <0.05.

[0167] conclusion As shown in Figures 5a and 5b, TQS-168 at 5 μM and 20 μM, respectively, can suppress TNFα production by LPS-stimulated BV-2 bone marrow cells compared to the control.

[0168] 5.5. Example 5 TQS-168 inhibits TNFα release by LPS-stimulated BV2 cells in vitro at concentrations between 0.3 μM and 10 μM (ELISA). TNFα ELISA procedure First, 100 μL of supernatant was removed from the cell plate and transferred to the dilution plate, which was then centrifuged at 216×g for 10 minutes to remove particulates. The dilution plate was either assayed immediately or aliquoted and stored at −20° C. or below; repeated freeze-thaw cycles were avoided.

[0169] A standard curve was then generated by first pipetting 900 ul of Calibration Diluent RD5K into the 700 pg / mL tube, followed by 200 μl of the appropriate Calibration Diluent into the remaining tubes. Stock solutions were used to produce the dilutions. The resulting tubes were then mixed thoroughly. The mouse TNFα standard (700 pg / mL) served as the high standard, and Calibration Diluent RD5T served as the zero standard at 0 pg / ml.

[0170] Assay diluent RD1-63 (50 uL) was then added to the center of each well and mixed before and during its use. 50 ul of either standard, control, or sample was then added to the center of each well and covered with an adhesive strip. The plate was then mixed for 1 minute and incubated at room temperature for 2 hours.

[0171] Each well was then aspirated and washed by filling each well with wash buffer (400 ul) five times. After the final wash, the remaining wash buffer was removed by aspirating or decanting. TNFα IL-6 conjugate (100 ul) was then added to each well, covered with an adhesive strip, incubated for 2 hours at room temperature, and washed and / or aspirated five times.

[0172] Substrate solution (100 uL) was then added to each well and incubated at room temperature in the dark for 30 min, followed by addition of 100 μl of stop solution and mixing. The optical density of each well was determined within 30 min by using a microplate reader set at 450 nm.

[0173] Measurement settings and parameters and data processing Plates were read at 450 nM and 570 nM on a Spectrostar Nano machine with built-in MARS data analysis.

[0174] Optimization parameters LPS stimulation was titrated to optimize the assay dynamic range.

[0175] In the TNFα ELISA, BV-2 cells were highly responsive to low concentrations of LPS. A concentration range from 0.1 ng / mL to 1,000 ng / mL was tested. LPS 0.3 ng / mL resulted in sufficient TNFα release from these cells (8-10 times background) after 22 h of stimulation without saturating the linear range of the ELISA detection system. If higher LPS concentrations are used to stimulate BV-2 cells, it is advised to keep the sample solution within the linear range of the detection system. The current TNFα protocol uses 10,000 cells per well of a 96-well plate. Cell counting titration may optimize the S / B ratio. Miniaturization from 96-well to 384-well is also feasible with this assay.

[0176] conclusion Figures 6a-6d show the dose response of TQS-168-mediated reduction of the proinflammatory cytokine TNFα release from microglial BV2 cells at 24 hours using ELISA. Figures 6a and 6c show the absolute (Figure 6a) and relative (Figure 6c) inhibition of TNFα secretion by BV2 cells stimulated with 0.3 ng / mL LPS. Figures 6b and 6d show the absolute (Figure 6b) and relative (Figure 6d) inhibition of TNFα secretion by BV2 cells stimulated with 1 ng / mL LPS.

[0177] In BV-2 cells treated with 0.3 ng / mL LPS, administration of TQS-168 inhibited TNFα production in a concentration-dependent manner, and in cells treated with 10 μM TQS-168, inhibition of up to approximately 25% was observed.

[0178] Similarly, in BV-2 cells treated with 1 ng / mL LPS, administration of TQS-168 suppressed TNFα production in a concentration-dependent manner, with the production being inhibited by approximately 35% in cells treated with 10 μM TQS-168.

[0179] 5.6. Example 6 TQS-168 inhibits LPS-induced TNF-α production by primary human bone marrow cells Research method Peripheral blood mononuclear cells (PBMCs) from four different healthy volunteers were used in this study. Fresh blood samples were collected at the Stanford Blood Center and processed for PBMC isolation using a Ficoll gradient. PBMC samples were stored in liquid nitrogen at -80°C for subsequent analysis of TNF-α production.

[0180] For TNF-α stimulation, frozen PBMC samples were thawed and rested at 37°C, after which the cells were stimulated with LPS 100ng / ml for 24 hours. DMSO (final dilution 1:1000) or various concentrations of TQS-168 in DMSO (final dilution 1:1000) were added to cell cultures of LPS-stimulated PBMC to evaluate the effect of T-168 on TNF-α production by human primary bone marrow cells. After 24 hours of stimulation, supernatant samples from the various conditions were collected and TNF-α concentrations in the supernatants were analyzed by cytometric bead array (CBA) assay according to the protocol from BD Biosciences. TNF-α was quantified by median fluorescence intensity reading (MFI). Samples were analyzed on an LSRII flow cytometer immediately after staining.

[0181] As shown in FIG. 7, in human PMBC cells treated with 100 ng / mL LPS, administration of TQS-168 at 5 μM and 20 μM resulted in a statistically significant inhibition of TNFα production compared to no treatment.

[0182] 5.7. Example 7 Tissue concentrations of TQS-168 after a single oral dose in mice Research method In the first experiment, a total of 3-4 male C57BL6 / J mice were administered a single dose of TQS-168 at 25 mg / kg by oral gavage. TQS-168 was prepared for oral gavage as a suspension in 0.5% methylcellulose in PBS.

[0183] Tissues were collected at various time points after administration and processed for LC-MS analysis of TQS-168 concentration. Tissues analyzed included plasma, brain, and liver. Animals were perfused extensively with 20 mL of ice-cold PBS prior to brain and liver collection to remove contaminating blood. Data are shown in Table 1 (TQS-168 concentration in ng / mL). [Table 1]

[0184] conclusion mean plasma C max The mean brain C max was 542.0 ng / ml, or 2.16 μM. The mean TQS-168 concentrations (ng / ml) are graphed in Figure 8 for plasma (Figure 8a), liver homogenate (Figure 8b), and brain homogenate (Figure 8c).

[0185] In the second experiment, mice were administered 50 mg / kg TQS-168 by oral gavage. In group 1, TQS-168 was prepared as a suspension in 0.5% methylcellulose in PBS at a concentration of 5.0 mg / mL. In group 2, TQS-168 was prepared as a solution in 10% polyethylene glycol (PEG) 400 / 30% Kleptose (Roquette) / 60% water at a concentration of 5.0 mg / mL.

[0186] Plasma exposure (ng / mL) is shown in Table 2 for Group 1 mice and Table 3 for Group 2 mice. [Table 2] [Table 3]

[0187] 9a-9c show the plasma concentrations of TQS-168 over time in Group 1 and Group 2 mice.

[0188] conclusion Plasma C was significantly increased after a single oral dose of 50 mg / kg, twice the dose previously demonstrated to suppress bone marrow-mediated inflammation and reduce disease severity in animal models of neurodegenerative diseases, including Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis (ALS). max 50 minutes C max Time to max ) was 2137ng / mL, or 8.54μM.

[0189] 5.8. Example 8 TQS-168 plasma and brain concentrations after a single oral dose in mice Mice were administered a single oral dose of 45 mg / kg of TQS-168, which was prepared as a solution in 10% polyethylene glycol (PEG) 400 / 30% Kleptose (Roquette) / 60% water at a concentration of 5.0 mg / mL.

[0190] As shown in Tables 4 and 5 below and in Figures 19A and 19B, concentrations in plasma and brain were measured at various time points and various pharmacokinetic parameters were calculated. [Table 4] [Table 5]

[0191] Brain-versus-plasma C of TQS-168 max The ratio was 1.842. The brain-to-plasma AUC ratio for TQS-168 was 1.814.

[0192] 5.9. Example 9 The metabolite TQS-621 inhibits LPS-stimulated release of proinflammatory cytokines from human PBMCs The major metabolites from the hepatic metabolism of TQS-168 after oral administration have been described. See Sun et al., Rapid Commun. Mass Spectrom. 32:480-488 (2018), incorporated herein by reference. The inventors synthesized the phase 1 metabolites from the hepatic metabolism of TQS-168 shown in Figure 20 and tested them for their ability to inhibit LPS-stimulated secretion of proinflammatory cytokines in vitro.

[0193] Briefly, 20,000 human PBMCs were aliquoted into each well. LPS was added at 1 ng / mL and cells were incubated for 24 hours in the presence of LPS and TQS-168 or one of its metabolites. Readouts were human IL-6 ELISA and human TNFα ELISA. For the assays, supernatants were diluted in culture medium. Supernatant dilution for IL-6 ELISA was 1:8 and for human TNFα ELISA was 1:2. Results are shown in Figures 21A and 21B.

[0194] Figure 21A shows the absolute inhibition of LPS-stimulated IL-6 secretion from PBMCs obtained from the first donor by the TQS-168 metabolite TQS-621. Figure 21B shows the relative inhibition of IL-6 inhibition expressed as percentage activity. The structure of TQS-621 is shown below in Formula II: [ka] It is shown as:

[0195] Figure 22A shows the absolute inhibition of LPS-stimulated IL-6 secretion from PBMCs obtained from a second donor by the TQS-168 metabolite TQS-621. Figure 22B shows the relative inhibition of IL-6 inhibition expressed as percentage activity.

[0196] Figure 23A shows the absolute inhibition of LPS-stimulated TNFα secretion from PBMCs of a first donor by TQS-168 and metabolite TQS-621. Figure 23B shows the relative inhibition of TNFα inhibition expressed as percentage activity. Figure 24A shows the absolute inhibition of LPS-stimulated TNFα secretion from PBMCs obtained from a second donor by TQS-168 and metabolite TQS-621. Figure 24B shows the relative inhibition of TNFα inhibition expressed as percentage activity.

[0197] The data indicate that the phase 1 metabolite TQS-621 is a potent inhibitor of LPS-stimulated IL-6 and TNFα secretion from human PBMCs, demonstrating that at least a portion of the therapeutic effects observed following oral administration of TQS-168 may be attributable to the activity of the active metabolite TQS-621.

[0198] 5.10. Example 10 Plasma concentrations of TQS-168 and its active metabolite TQS-621 following a single oral dose of TQS-168 in mice Male C57BL / 6 mice were administered a single dose of TQS-168 at 50 mg / kg using one of three formulations: Group 1 - 5.0 mg / ml suspension in 0.5% methylcellulose in PBS; Group 2 - 5.0 mg / ml solution in PEG400 (Fluka) 10% / Kleptose (Roquette) 30% / sterile water 60%; Group 3 - 5.0 mg / ml suspension in Labrafil M 1944 CS (Gattefosse) 30% / Masine CC (Gattefosse) 5% / Miglyol 812N 10% / sterile water 55%.

[0199] Plasma concentrations of TQS-168 and TQS-621 were measured over time.

[0200] Table 6 presents the TQS-168 plasma concentration data for Group 1 mice; Table 7 presents the TQS-168 plasma concentration data for Group 2 mice; Table 8 shows the TQS-168 plasma concentration data for Group 3 mice. [Table 6-1] [Table 6-2] [Table 7] [Table 8] The solution formulation, Group 2, had a higher TQS-168C than the suspension formulations, Group 1 (1121 ng / mL) (4.5 μM) and Group 3 (611 ng / mL) (2.4 μM). max (1950 ng / mL) (7.9 μM) and had a higher exposure (AUC inf =11404hr * The results are plotted in Figures 25A (group 1), 25B (group 2), and 25C (group 3).

[0201] Table 9 presents the TQS-621 plasma concentration data for Group 1 mice; Table 10 presents the TQS-621 plasma concentration data for Group 2 mice; Table 11 shows the TQS-168 plasma concentration data for Group 3 mice. [Table 9] [Table 10] [Table 11-1] [Table 11-2]

[0202] TQS-621 plasma C in mice in Group 2 given the solution formulation max was 449 ng / mL, and the C of metabolite TQS-621 relative to parent TQS-168 was 0.152. max ratio, and AUC of 0.136 last yielded the ratio.

[0203] The results are plotted in Figures 26A (group 1), 26B (group 2), and 26C (group 3).

[0204] 5.11. Example 11 Brain TQS-621 concentrations after a single oral dose of TQS-168 in mice TQS-168 was prepared as a 4.5mg / ml solution in PEG400 10% / Kleptos 30% / sterile water 60%. A single oral dose of 45mg / kg was administered. Concentrations of TQS-621 were measured over time in plasma and brain. The data are shown in Tables 12 and 13 below. [Table 12] [Table 13]

[0205] 5.12. Example 12 Plasma and brain concentrations of TQS-168 after a single oral dose in rats Research method Both male (n=3) and female (n=3) Sprague-Dawley rats were administered a single dose of TQS-168 by oral gavage at 50, 150, or 500 mg / kg TQS-168. TQS-168 was prepared as a suspension in 0.5% methylcellulose in PBS.

[0206] Plasma samples were collected at 30, 60, 120, 240, and 1440 minutes after dosing. Brain samples were collected in some animals at 60, 240, and 1440 minutes after a single oral dose of 500 mg / kg TQS-168. Samples were processed for LC-MS analysis of TQS-168 concentration.

[0207] The data are expressed as the concentration (ng / ml) of TQS-168 in plasma (Figure 12a) and brain homogenates (Figure 12b). Figures 12a and 12b show that TQS-168 was detected in both plasma and brain tissue of treated rats. The highest concentration of TQS-168 (C max ) was detected in both plasma and brain 240 min after oral administration of TQS-168.

[0208] Using PKSolver, C from plasma tissue max and AUC 0-t The values ​​were calculated (Comput Methods Programs Biomed. 2010 Sep;99(3):306-14. doi: 10.1016 / j.cmpb.2010.01.007. Epub 2010 Feb 21) and the underlying data are plotted in Fig. 12a. The results are shown in Fig. 13 (C max ) and 14 (AUC), and the values ​​are summarized in Table 14. [Table 14]

[0209] conclusion Following single oral doses of 50, 150, and 500 mg / kg in rats, TQS-168 exhibited dose-dependent C max and AUC were detected.

[0210] 5.13. (Example 13) Plasma concentrations of TQS-168 after a single intravenous dose in mice Research method Three male 7-9 week old CD-1 mice from Lingchang were treated intravenously with 0.5mg / kg TQS-168. TQS-168 was prepared in a solution of DMAC 31.6% + ethanol 36.8% + propylene glycol 31.6%. Blood was collected at 0.5, 3, 10, 30, 60, 120, 240, 480 and 720 minutes after a single dose of TQS-168 and then processed. Blood samples were collected into K2EDTA tubes via saphenous vein puncture, centrifuged at 4600 rpm for 5 minutes at 4°C, and plasma was collected and stored below -20°C before being analyzed by LC-MS for TQS-168 concentration. Data are expressed as the concentration of TQS-168 per volume of plasma and the average concentration (ng / mL) as shown in Figure 10 (individual mice) and Figure 11 (average), respectively.

[0211] conclusion Following intravenous administration of 0.5 mg / kg TQS-168 in mice, TQS-168 was detected in plasma with a terminal elimination half-life of 0.14 hours (8.4 minutes).

[0212] 5.14. Example 14 Detection of TQS-168 in plasma after a single intravenous administration in rats Research method A total of three male 7-9 week old Sprague Dawley rats from Vital River were administered a single intravenous dose of TQS-168 at 0.5 mg / kg in a volume of 0.5 mL / kg. TQS-168 was prepared in a solution of DMAC 31.6% + ethanol 36.8% + propylene glycol 31.6%.

[0213] Blood samples were collected at 0.5, 3, 10, 30, 60, 120, 240, 480 and 720 minutes after a single dose of TQS-168 and then processed. Blood samples were collected via saphenous vein puncture into K2EDTA tubes, centrifuged at 4600 rpm for 5 minutes at 4°C, and plasma was collected and stored below -20°C before being analyzed by LC-MS for TQS-168 concentration. Data are presented as plasma TQS-168 concentration (ng / mL) and are graphed in Figure 15 (individual rats) and Figure 16 (average).

[0214] conclusion TQS-168 was detected in plasma after administration of a single intravenous dose of 0.5 mg / kg in rats. The half-life was 0.162 hours (9.72 minutes) with a clearance rate of 9.29 (L / hr / kg).

[0215] 5.15. Example 15 Detection of TQS-168 in plasma after a single intravenous administration in dogs Research method A total of three male 1- to 3-year-old beagle dogs from Beijing Marshall Biotechnology, Ltd. were treated intravenously with 0.5 mg / kg TQS-168 at 0.5 ml / kg. TQS-168 was prepared in a solution containing 31.6% DMAC, 36.8% ethanol, and 31.6% propylene glycol.

[0216] Blood samples were collected at 0.5, 3, 10, 30, 60, 120, 240, 360, 480, 720 and 1440 minutes after a single dose of TQS-168 and then processed. Blood samples were collected into K2EDTA tubes, centrifuged at 4600 rpm for 5 minutes at 4°C, and plasma was collected and stored below -20°C before being analyzed by LC-MS for TQS-168 concentration. Data are expressed as concentration of TQS-168 per plasma volume (ng / mL) and are shown in Figures 17 and 18.

[0217] 5.16. Example 16 Part 1: SAD Phase 1 Study - A double-blind, randomized study of subjects receiving single ascending doses of TQS-168 or placebo. 5.16.1. Study Design We conducted a double-blind, randomized, placebo-controlled clinical trial to characterize and compare the pharmacokinetic (PK) profiles of TQS-168 and its metabolite TQS-621 following single ascending doses of TQS-168 delivered in three different formulations, or placebo, in healthy subjects.

[0218] Subjects: This randomized, double-blind, placebo-controlled, Phase 1 single ascending dose [SAD] study enrolled subjects with a body mass index (BMI) of 18.0 to 32.0 kg / m2 as measured at screening. 2 The study was conducted on healthy male subjects aged 18-55 years with pulmonary embolism and chronic obstructive pulmonary disease (COPD). All subjects weighed at least 55 kg at screening. Key criteria for exclusion were subjects with current evidence of SARS-CoV-2 infection, clinical manifestations of significant cardiovascular, renal, hepatic, skin, chronic respiratory or gastrointestinal disease, or aspartate aminotransferase (AST) or alanine aminotransferase (ALT) >1.5 times the upper limit of normal (ULN). Subjects were recruited at a single site in the UK. Each subject provided written informed consent.

[0219] Study Design: The study was conducted in multiple cohorts, each containing a minimum of seven subjects. In cohorts 1-3 of part 1, subjects received a single oral dose of TQS-168 methylcellulose (MC) suspension, or spray-dried dispersion (SDD) suspension, or hot melt extrudate (HME) suspension in the fasted state, or placebo. Subjects were assigned to study treatment in a ratio of 6 TQS-168 to 2 placebo per cohort. Subjects in cohort 1 received regimen A, TQS-168 MC 60 mg. Subjects in cohort 2 received regimen B, TQS-168 180 mg. Cohort 3 was divided into three separate periods, whose subjects were single-regimen recipients. Subjects in cohort 3, period 1, received regimen C, TQS-168 MC 540 mg suspension on day 1. The same subjects in Cohort 3, Period 2 received Regimen D, TQS-168 SDD 180 mg suspension on Day 1, and in Part 1, Cohort 3, Period 2, the same subjects received Regimen E, TQS-168 HME 180 mg suspension on Day 3. See Table 15. [Table 15-1] [Table 15-2]

[0220] For cohorts 1-3, period 1, the screening period was up to 4 weeks. After confirming eligibility, subjects within each cohort were randomly assigned to receive either active (TQS-168) or placebo treatment. Note that this is the first time TQS-168 is administered to humans, and therefore, a sentinel dosing design is followed. Each cohort was divided into a sentinel group and a main group. The sentinel group consisted of the first two subjects in each cohort. They were administered before the remaining subjects, the main group. Only after a positive review of the sentinel group's safety data up to 24 hours after dosing were the main group subjects within a cohort dosed. The randomization schedule was performed such that one of the subjects dosed on day 1 was given the TQS-168 suspension and one was given the placebo. The protocol ensured that all treatments were taken once daily after an overnight fast (fasting for ≥10 hours) except for the cohort in which food effects were evaluated. In cohorts 1-3, period 1, subjects were admitted the morning before dosing (day -1) and remained in the facility until 48 hours after dosing (day 3) at which time they were discharged. In cohort 3, period 2, subjects were admitted the morning before dosing (day -1) and remained in the facility until 48 hours after dosing (day 5) at which time they were discharged. Depending on the assigned regimen, subjects were dosed on the morning of day 1 either in the fasted state after an overnight fast (≥10 hours) or in the fed state after a high-fat breakfast given 30 minutes before dosing.

[0221] A safety study was conducted after dosing in cohorts 1-3, period 1 and before dosing subjects in cohort 3, period 2. The study concluded that it was safe to administer the SDD and HME formulations to subjects in similar dosages as the MC formulation. As a result, Sentinel was not required in cohort 3, period 2.

[0222] In cohorts 4-6, each subject received a single oral dose of TQS-168 SDD suspension or placebo in a fed or fasted state. Subjects were randomly assigned to study treatment vs. placebo in a 5:1 ratio per cohort. Cohort 4 received regimen F, in which 90 mg of TQS-168 was administered to fed subjects. Cohort 5 received regimen G, in which 90 mg of TQS-168 was administered to fasted subjects. Cohort 6 received regimen H, in which 270 mg of TQS-168 was administered to fasted subjects. Per protocol, subjects were dosed on the morning of Day 1 either in the fasted state after an overnight fast (≥10 hours) or in the fed state after a high-fat breakfast given 30 minutes prior to dosing. See Table 16.

[0223] Safety was assessed continuously throughout the study by monitoring adverse events and use of concomitant medications, electrocardiograms (ECGs), vital signs, laboratory safety assessments, and physical examinations. Blood samples for pharmacokinetic assessments were collected from each subject beginning on Day -1, prior to (within 1 hour) each dose, and at intervals throughout the study up to 48 hours post-dose, if applicable. In cohorts 1-5, no individual subject achieved the prestudy designed Cmax of 305 ng / mL and 2750 ng / mL. * AUC in h / mL (0-24) No doses were used that were predicted to exceed the exposure cap of 112.1 ng / mL and 596.9 ng / mL, respectively. * h / mL. [Table 16]

[0224] Safety was assessed continuously throughout the study by monitoring adverse events and use of concomitant medications, electrocardiograms (ECGs), vital signs, laboratory safety assessments, and physical examinations in all subjects in cohorts 1 to 3. Blood samples for pharmacokinetic assessments were collected from each subject beginning on day -1, prior to each dose (within 1 hour), and at intervals throughout the study up to 48 hours after dosing, if applicable.

[0225] Pharmacokinetic Evaluation: Blood samples for plasma PK analysis were collected at regular time intervals. Venous blood samples were collected from subjects by trained members of the clinical team. Pre-dose samples were collected within 1 hour pre-dose. Timestamp 0 to 1 hour post-dose samples were collected within ±2 minutes from the reference post-dose sampling time. Timestamp 1.5 to 12 hours post-dose samples were collected within ±10 minutes from the reference post-dose sampling time. Timestamp 16 to 48 hours post-dose samples were collected within ±30 minutes from the reference post-dose sampling time. Samples were collected in appropriate containers and processed to isolate plasma. PK analysis was performed on plasma samples using validated biochemical analytical methods.

[0226] Statistical Analysis: The sample size for the study was selected based on practical considerations and experience from previous studies of similar design. The number of subjects in each cohort was deemed adequate to evaluate the primary objective of each study. Pharmacokinetic parameters were determined by non-compartmental techniques using WinNonlin software version 8.0 or higher (Certara USA. Inc., USA). All data were enumerated and summarized by subject group using descriptive statistics. All statistical analyses were performed using SAS version 9.4 or higher. 5.16.2. TQS-168 Methylcellulose (MC) Powder Suspension Formulation A methylcellulose (MC) powder suspension formulation of 2-(4-tert-butylphenyl)-1H-benzimidazole (TQS-168; compound of formula I) was prepared by reconstitution as a suspension in the methylcellulose vehicle formulation of Table 17 ("Vehicle Formulation"). [Table 17-1] [Table 17-2]

[0227] A vehicle formulation was prepared by heating water (1986 g) to 80° C. (+5° C.) and then adding methylcellulose (10 g) and stirring for 30 minutes or more until the methylcellulose was completely dispersed. Sodium dodecyl sulfate (2 g) and 30% simethicone emulsion (2 g) were then added and the mixture was stirred until a semi-transparent white / off-white, slightly viscous suspension, free-form particulates were formed. The pH of the resulting vehicle formulation was 5.3 (target pH was 6.0+ / -3.0).

[0228] General Procedure for Reconstituting Compounds of Formula I in Vehicle Formulations of Table 17 The required amount (e.g., 60 mg to 1000 mg) of 2-(4-tert-butylphenyl)-1H-benzimidazole (compound 1; TQS-168) was weighed into a vial. A vehicle formulation (100 mL) was added to the vial containing the compound of formula I to obtain a compound of formula I methylcellulose (MC) powder suspension formulation.

[0229] 5.16.3. Methylcellulose (MC) Results Healthy male subjects were administered a single oral dose of TQS-168 methylcellulose (MC) powder in the fasted state at one of the following doses: 60 mg, 180 mg, or 540 mg. Plasma concentrations of TQS-168 and metabolite TQS-621 were measured over time, and key pharmacokinetic parameters were determined.

[0230] Tables 18 and 19 present the geometric means of important pharmacokinetic parameters of TQS-168 and metabolite TQS-621 in subjects following oral administration of TQS-168. [Table 18] [Table 19-1] [Table 19-2]

[0231] TQS-168 Single Ascending Dose (SAD) PK Profile: Cohort 3, Period 1 received TQS-168 540 mg (Regimen C) with a peak TQS-168C of 323 ng / mL (1.29 μM) max Cohort 2 received 180 mg of TQS-168 (regimen B) and 53.1 ng / mL (0.21 μM) of TQS-168C. max Cohort 1 received TQS-168 60 mg (regimen A) and 26.7 ng / mL (0.11 μM) TQS-168C max The results are plotted in Figure 27. Cohort 3, Period 1 showed a significantly higher mean mean mean than Cohort 2 (AUC 0-24 =163hr * ng / m, AUC 0-inf =180hr * ng / mL) and cohort 1 (AUC 0-24 =71.2hr * ng / mL, AUC 0-inf =72.7hr * ng / mL) than 0-24 =1440hr * ng / mL, AUC 0-inf =1600hr * The concentrations (ng / mL) are also shown.

[0232] Metabolite TQS-621 PK profile: Cohort 3, Period 1 had higher metabolite TQS-621C than Cohort 2 (199 ng / mL) (0.75 μM) and Cohort 1 (65.3 ng / mL) (0.25 μM) max (1110 ng / mL) (4.17 μM). The results are plotted in FIG. 28. Similarly, cohort 3 showed a significantly higher AUC 0-24 =1210hr * ng / mL, AUC inf =1350hr * ng / mL) and cohort 1 (AUC 0-24 =383hr * ng / mL, AUC inf =410hr * ng / mL) than metabolite exposure (AUC 0-24=10800hr * ng / m, AUC inf =12700hr * The concentrations (ng / m) are also shown.

[0233] In the study, TQS-168(C max ) and TQS-168 exposure (AUC (0-24) and AUC (0-inf) ) appeared to increase somewhat inversely proportionally with dose after single doses of 60 and 180 mg of TQS-168. However, at doses of TQS-168 between 180 mg and 540 mg, the C max , AUC (0-24) and AUC (0-inf) Note that C increased in a supra-proportional manner, 6.1, 8.8, and 9.5-fold with a 3-fold increase in dose, respectively. Looking at the entire dose range from 60 mg to 540 mg, C max , AUC (0-24) and AUC (0-inf) were all increased super-proportionally by 12.1-fold, 20.2-fold, and 21.2-fold, respectively. See Figures 27, 29-31.

[0234] For the metabolite TQS-621, plasma C max , AUC (0-24) and AUC (0-inf) appeared to increase in a dose-proportional manner after single doses of 60–180 mg of TQS-168. At doses between 180 and 540 mg, C max , AUC (0-24) and AUC (0-inf) increased superproportionately with 5.6, 8.9, and 9.4-fold increases with each 3-fold increase in dose, respectively. Furthermore, plasma C increased significantly with each 9-fold increase in dose from 60 mg to 540 mg across the entire dose range. max , AUC (0-24) and AUC (0-inf) increased super-proportionally by 17.0, 28.2 and 31.0 fold, respectively. See Figures 28-31.

[0235]

[0236] After administration of 60 mg, the maximum plasma TQS-168 concentration was 1.0 mg / kg, the median T max Maximum plasma metabolite TQS-621 concentrations occurred between 0.5 and 2 hours after dosing at a median T of 2.5 hours after dosing. max occurred between 1 and 4 hours after administration.

[0237] After administration of 180 mg of TQS-168, maximum plasma concentrations of TQS-168 occurred between 1 and 4 hours post-dose. Median T max The maximum plasma concentration of the metabolite TQS-621 was observed at a median of 1.5 hours after administration. max occurred between 1 and 4 hours after administration.

[0238] After administration of 540 mg of TQS-168, the maximum plasma concentration of TQS-168 was observed at a median of 2.25 hours post-dose. max The metabolite TQS-621 had a median T of 4 hours after dosing. max The maximum plasma concentration was observed between 2 and 12 hours after administration.

[0239] At all administered doses of TQS-168, concentrations of the metabolite TQS-621 exceeded those of TQS-168 after a short lag.

[0240] The geometric mean terminal half-lives of TQS-168 after 60 mg, 180 mg, and 540 mg doses were dose-dependent at 3.06, 7.36, and 10.1 hours, respectively. The geometric mean terminal half-lives of the metabolite TQS-621 after 60 mg, 180 mg, and 540 mg doses were 4.89, 9.10, and 11.3 hours, respectively. It is noted that for all doses of TQS-168, plasma concentrations of TQS-168 were quantifiable from 0.5 hours post-dose and remained quantifiable until the last sampling time of 48 hours post-dose. Concentrations of TQS-621 were also quantifiable from 0.5 hours post-dose and remained quantifiable until the last sampling time of 48 hours post-dose.

[0241] 5.16.4. Spray-dried dispersion (SDD) formulations and oral suspensions 5.16.4.1 Preparation of SDD formulations of 2-(4-tert-butylphenyl)-1H-benzimidazole (compound of formula I) A spray-dried dispersion (SDD) of 2-(4-tert-butylphenyl)-1H-benzimidazole (compound of formula I; TQS-168) having the composition shown in Table 20 was prepared by spray drying the feedstock formulation shown in Table 21. [Table 20] [Table 21]

[0242] 5.16.4.2 Manufacturing Procedures for Spray-Dried Feedstock Blends The compound of formula I (45.0 g) was slowly added to 2-propanol (1791.1 g) with stirring, placed under a homogenizer (Silverson SL2 homogenizer) and stirred for 5 minutes or more until the compound of formula I was completely dissolved. The reaction mixture was then removed from the homogenizer, polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (Soluplus) (60.0 g) was slowly added with stirring, placed back in the homogenizer and stirred for 10 minutes or more until the Soluplus was completely dissolved. The reaction mixture was then removed from the homogenizer, amorphous silicon dioxide (Syloid® 244 FP) was slowly added with stirring, placed back in the homogenizer and stirred for an additional 15 minutes or more until the amorphous silicon dioxide was completely dispersed. The resulting suspension is referred to herein as the "feedstock blend."

[0243] 5.16.4.3 Manufacturing Procedures for Spray-Dried Dispersion (SDD) Formulations of Compounds of Formula I A spray dryer unit (ProCepT 4M8 Spray Dryer) was equipped with a compressed air supply. Once the outlet temperature had stabilized, the feed pump was started and 2-propanol (blank solution) was sprayed as a fine spray through the nozzle into the collection chamber. The spray dryer parameters were adjusted to achieve a feed rate within the ranges shown in Table 22 below.

[0244] The feed blend was stirred under the homogenizer at an appropriate speed to maintain a uniform dispersion without creating air bubbles.The feed blend was then sprayed as a fine spray through the nozzle into the collection chamber of the spray dryer unit (ProCepT 4M8 SprayDryer, prepared with blank solution and using parameters as outlined in Table 3), where the solvent quickly evaporated to produce particles containing the compound of formula I, polyvinylcaprolactam-polyvinylacetate-polyethyleneglycol graft copolymer (Soluplus) and silicon dioxide (Syloid® 244 FP) (SDD formulation of compound 1).Once all the feed blend was sprayed and collected, the feed blend was replaced with 2-propanol (blank solution) and sprayed through the nozzle of the spray dryer for 5 minutes or more to allow any remaining "feed blend" in the air stream to be collected. [Table 22] 5.16.4.4 Oral suspension of a spray-dried dispersion (SDD) formulation of 2-(4-tert-butylphenyl)-1H-benzimidazole (Compound 1) A spray dried dispersion (SDD) of 2-(4-tert-butylphenyl)-1H-benzimidazole (Compound 1) (e.g., 60-1000 mg) having the composition shown in Table 20 was reconstituted as an oral suspension in 100 g of vehicle consisting of PEG300 (10 g), glycerol monocaprylocaprate (Capmul MCM, 0.40 mg) in sterile water for irrigation (qs to 100 g).

[0245] 5.16.5. SDD results Healthy male subjects were administered a single oral dose of 90 mg, 180 mg, or 270 mg of TQS-168 SDD powder oral suspension in a fed or fasted state. Plasma concentrations of TQS-168 and metabolite TQS-621 were measured over time, and key pharmacokinetic parameters were determined.

[0246] Tables 23 and 24 present key pharmacokinetic parameters of TQS-168 and metabolite TQS-621 in subjects following oral administration of the TQS-168 SDD formulation. [Table 23] [Table 24-1] [Table 24-2]

[0247] TQS-168 Single Ascending Dose (SAD) PK Profile: Cohort 3, part 2 received regimen D, 180 mg TQS-168 SDD powder for oral suspension in the fasted state, resulting in a TQS-168 Cmax of 218 ng / mL (0.87 μM). Cohort 4 received regimen F, 90 mg TQS-168 SDD powder for oral suspension in the fed state, resulting in a TQS-168 Cmax of 47.1 ng / mL (0.19 μM). Cohort 5 received regimen G, 90 mg TQS-168 SDD powder for oral suspension in the fasted state, resulting in a TQS-168 Cmax of 111 ng / mL (0.44 μM). Cohort 6 received Regimen H, 270 mg of TQS-168 SDD powder for oral suspension in the fasted state, which resulted in the highest TQS-168 Cmax of 237 ng / mL (0.95 μM). Results are plotted in Figure 44. Recipients of Regimen H had a significantly higher TQS-168 Cmax than recipients of Regimen D (AUC 0-24 =608hr * ng / mL, AUC 0-inf =621hr * ng / mL), Regimen G (AUC 0-24=192hr * ng / mL, AUC 0-inf =195hr * ng / mL) and Regimen F (AUC 0-24 =168hr * ng / mL, AUC 0-inf =171hr * ng / mL) recipients with higher exposure (AUC 0-24 =736hr * ng / mL, AUC 0-inf =836hr * The concentrations (ng / mL) are also shown.

[0248] Metabolite TQS-621 PK Profile: Regimen D (180 mg TQS168) resulted in a higher metabolite TQS-621 Cmax (742 ng / mL, 2.79 μM) than Regimen H (621 ng / mL, 2.33 μM), Regimen G (214 ng / mL, 0.80 μM) and Regimen F (122 ng / mL, 0.46 μM). Results are plotted in Figure 45. Similarly, recipients of Regimen D had a higher Cmax (AUC 0-24 =4250hr * ng / mL, AUC 0-inf =4950hr * ng / mL), Regimen G (AUC 0-24 =1170hr * ng / mL, AUC 0-inf =1200hr * ng / mL) and Regimen F (AUC 0-24 =848hr * ng / mL, AUC 0-inf =901hr * ng / mL) recipients had higher metabolite exposure (AUC 0-24 =5110hr * ng / mL, AUC 0-inf =5170hr * The concentrations (ng / mL) are also shown.

[0249] Comparing the administration of 270 mg of TQS-168 SDD powder for oral suspension in the fasted state (Regimen H) to the administration of 90 mg of TQS-168 SDD powder for oral suspension in the fasted state (Regimen G) revealed that a 3-fold increase in dose resulted in a 2.14- and 4.29-fold increase in TQS-168 Cmax and AUC(0-inf), and a 2.90- and 4.13-fold increase in TQS-621 Cmax and AUC(0-inf). See Figures 38-39.

[0250] In a notable comparison of administration of 90 mg of TQS-168 SDD powder for oral suspension in the fasted state (Regimen G) versus the same administration in the fed state (Regimen F), administration in the fasted state demonstrated an increase in TQS-168 Cmax and AUC(0-inf) of approximately 136% and 14%, and TQS-621 Cmax and AUC(0-inf) of 75% and 33%. See Figures 35-36.

[0251] Comparing 270 mg of TQS-168 SDD powder for oral suspension administered in the fasted state (Regimen H) with 180 mg of TQS-168 administered in the fasted state (Regimen D) revealed an increase in TQS-168 Cmax and AUC(0-inf) of approximately 8.7% and 26%, but a decrease in TQS-621 Cmax and AUC(0-inf) of approximately 16% and 4.3%. See Figures 38 and 46.

[0252] Following oral administration of 180 mg of TQS168SDD in the fasting state, maximum plasma TQS-168 concentrations occurred between 0.5 and 2 hours post-dose, with a median Tmax of 1 hour. Maximum plasma metabolite TQS-621 concentrations occurred between 0.5 and 2 hours post-dose, with a median Tmax of 2 hours post-dose. max This occurred between 1.5 and 3.0 hours post-dose. See Figures 46A-B.

[0253] After oral administration of 90 mg of TQS-168 SDD in the fed state in Regimen F, the maximum plasma TQS-168 concentration was 1.5 hours post-dose at a median of 1. maxThe maximum plasma concentration of the metabolite TQS-621 was 0.5 to 3 hours after dosing, with a median T of 3.5 hours after dosing. max and occurred between 1.5 and 6.0 hours post-dose. See Figures 34A-B.

[0254] Following oral administration of 90 mg of TQS-168 SDD in the fasted state in Regimen G, maximum plasma concentrations of TQS-168 occurred between 0.5 and 2.0 hours post-dose. Median T max The maximum plasma concentration of the metabolite TQS-621 was observed at a median of 1.25 hours post-dose. max and occurred between 1.0 and 3.0 hours post-dose. See Figures 37A-B.

[0255] Regimen H, following oral administration of 270 mg of TQS-168 SDD in the fasting state. Maximum plasma concentrations of TQS-168 were observed at a median T of 1.50 hours post-dose. max The metabolite TQS-621 had a median T of 2 hours after dosing. max and showed maximum plasma concentrations between 1.5 and 4 hours after administration. See Figures 38-39.

[0256] Terminal half-life (T 1 / 2 ) were dose-dependent at 3.52, 4.85, 5.64, and 10.4 hours, respectively. The terminal half-lives of the TQS-168 metabolite TQS-621 at doses of 90 mg fed, 90 mg fasted, 180 mg fasted, and 270 mg fasted were 5.17, 4.79, 7.12, and 10.4 hours, respectively. It is noted that for all doses of TQS-168, plasma concentrations of TQS-168 were quantifiable from 0.5 hours post-dose and remained quantifiable until the final sampling time of 48 hours post-dose. Concentrations of TQS-621 were also quantifiable from 0.5 hours post-dose and remained quantifiable until the final sampling time of 48 hours post-dose.

[0257] 5.16.6. HME preparations and oral suspensions 5.16.6.1 Hot Melt Extrudate (HME) Formulation of 2-(4-tert-butylphenyl)-1H-benzimidazole (Compound 1) HME Preparation A hot melt extrudate (HME) formulation of 2-(4-tert-butylphenyl)-1H-benzimidazole (compound of formula I; TQS-168) having the composition shown in Table 24 was prepared as shown below. [Table 25]

[0258] The required amount of compound 1, polyvinylcaprolactam-polyvinylacetate-polyethyleneglycol graft copolymer (Soluplus), povidone (Kollidon 17PF) and copovidone (Kollidon VA64) from Table 25 was weighed, passed through an 850 μm sieve and transferred to a 2L blender shell. The resulting blender shell was secured to a blender (Pharmatech blender) and blended for 20 minutes, then added to a double polyethylene (PE) bag ("Formula I compound blend") and transferred to the HME containment system. The chiller unit was connected to the HME, and once the chiller temperature reached 15°C, the extrusion process was started using the parameters outlined in Table 26. The Formula I compound blend was added to the feeder to fill it approximately 3 / 4, and the extrudate was collected and discarded for approximately the first 5 minutes of the extrusion process. The feeder was refilled to maintain approximately 50% volume of the feeder throughout the process, and extrusion continued until all of the compound of Formula I blend had been extruded and collected ("compound of Formula I HME extrudate"). [Table 26-1] [Table 26-2]

[0259] The collected compound of formula I HME extrudate was added to a U5 Quadro mill (set at 457 (mm) screen size and 5000 RPM impeller speed) until all extrudate passed through the 457 mm screen to obtain finely divided granules of compound of formula I. The finely divided granules of compound 1 were then sieved using a 300 micron sieve and transferred to a blender shell (Pharmatech 2L blender shell). The resulting blender shell was secured to the blender (Pharmatech blender), blended for 5 minutes, and collected.

[0260] 5.16.6.2 Oral Suspension of Hot Melt Extrudate (HME) Formulation of 2-(4-tert-butylphenyl)-1H-benzimidazole (Compound of Formula I) A hot melt extrudate (HME) of 2-(4-tert-butylphenyl)-1H-benzimidazole (compound of formula I) (e.g., 60-1000 mg) having the composition shown in Table 25 was reconstituted as an oral suspension in 100 mL of the vehicle Ora-Blend SF® (purified water, sucrose, glycerin, sorbitol, flavorings, microcrystalline cellulose, sodium carboxymethylcellulose, xanthan gum, carrageenan, citric acid, sodium phosphate, simethicone, potassium sorbate, and methylparaben), a commercially available oral suspension vehicle manufactured by Perrigo Pharmaceuticals.

[0261] 5.16.7. HME results Healthy male subjects were administered a single oral dose of 180 mg of TQS-168 HME powder oral suspension in the fasted state (Regimen E). Plasma concentrations of TQS-168 and metabolite TQS-621 were measured over time, and key pharmacokinetic parameters were determined.

[0262] Tables 27 and 28 present the geometric means of key pharmacokinetic parameters of TQS-168 and metabolite TQS-621 in subjects following oral administration of the TQS-168 SDD formulation. [Table 27] [Table 28]

[0263] TQS-168 Single Ascending Dose (SAD) PK Profile: Cohort 3, Period 2 received 180 mg of TQS-168 HME powder in oral suspension in the fasted state in Regimen E. The single dose resulted in a TQS-168 Cmax of 123 ng / mL (0.49 μM) and a PK profile of 358 hr * The AUC in ng / mL was obtained. The data are plotted in Figures 47A-B.

[0264] Metabolite TQS-621 PK profile: Regimen E achieved metabolite TQS-621 Cmax and 3090hr of 481ng / mL (1.81μM). * AUC in ng / mL 0-24 This is illustrated in Figures 47A-B.

[0265] 5.16.8. Comparison of PK outcomes for MC vs. SDD vs. HME In part 1 of this treatment, subjects were given 180 mg of TQS-168 in the fasted state in the following formulations: methylcellulose (MC), spray dried dispersion (SDD) powder in oral suspension, or hot melt extrudate (HME) powder in oral suspension. Plasma concentrations of TQS-168 and metabolite TQS-621 were measured over time and key pharmacokinetic parameters were determined as previously shown. For convenience, results are compared in Tables 29-31. See Figures 45A-B. [Table 29] [Table 30-1] [Table 30-2] [Table 31]

[0266] Subjects receiving 180 mg of TQS-168 MC powder for oral suspension (Regimen B) demonstrated quantifiable plasma concentrations of TQS-168 beginning 0.5 hours post-dose and remaining quantifiable from 10 to 48 hours post-dose. TQS-621 concentrations were also quantifiable beginning 0.5 hours post-dose and remaining quantifiable from 24 to 48 hours post-dose.

[0267] Maximum plasma TQS-168 concentrations occurred between 1 and 4 hours post-dose, with a median Tmax of 1 hour. 1 / 2 The geometric mean (CV%) Cmax and AUC(0-inf) values ​​were 53.1 ng / mL (55%) and 180 ng / mL, respectively. * h / mL (53.1%).

[0268] Maximum plasma TQS-621 concentrations occurred between 1 and 4 hours post-dose, with a median Tmax of 1.5 hours post-dose. 1 / 2 The geometric mean (CV%) Cmax and AUC(0-inf) values ​​were 199 ng / mL, 10.2 ng / mL, and 10.3 ng / mL, respectively. * hr / mL (33.5%) and 1350ng * h / mL (45.1).

[0269] Following administration of 180 mg of TQS-168 SDD powder for oral suspension (Regimen D), plasma concentrations of TQS-168 were quantifiable beginning at 0.5 hours post-dose and remained quantifiable between 24 and 36 hours post-dose. It is noted that one subject exhibited a quantifiable TQS-168 plasma concentration pre-dose as a result of some carryover from the prior dosing regimen. TQS-621 concentrations were quantifiable from pre-dose in all subjects as a result of some carryover from the prior dosing regimen. This remained quantifiable until the last sampling time point 48 hours post-dose (Day 5). It is noted that all quantifiable pre-dose concentrations were less than 5% of Cmax.

[0270] Maximum plasma TQS-168 concentrations occurred between 0.5 and 2 hours post-dose, with a median Tmax of 1 hour. 1 / 2 The maximum plasma TQS-621 concentrations occurred between 1.5 and 3 hours after dosing, with a median Tmax of 2 hours. The resulting geometric mean T 1 / 2 was 7.12 hours.

[0271] Geometric mean Cmax, AUC(0-last) and AUC(0-inf) of TQS-168 after administration of 180 mg of SDD powder for oral suspension were increased by 4.11, 3.64 and 3.45 fold, respectively, when compared to its MC counterpart (Regimen B). Geometric mean Cmax, AUC(0-last) and AUC(0-inf) of TQS-621 after administration of 180 mg of SDD powder for oral suspension were increased by 3.73, 3.87 and 3.83 fold, respectively, when compared to its MC counterpart (Regimen B).

[0272] Following administration of 180 mg of TQS-168 HME powder for oral suspension (Regimen E), subjects demonstrated quantifiable plasma concentrations of TQS-168 beginning 0.5 hours post-dose and remaining quantifiable between the last sampling time of 16 and 48 hours post-dose (Day 5). TQS-621 concentrations were also quantifiable beginning 0.5 hours post-dose in all subjects and remained quantifiable until the last sampling time of 48 hours post-dose.

[0273] Maximum plasma TQS-168 concentrations occurred between 1 and 1.5 hours post-dose, with a median Tmax of 1.5 hours post-dose. 1 / 2 The maximum plasma TQS-621 concentrations occurred between 1.5 and 4 hours after dosing, with a median Tmax of 2 hours. The resulting geometric mean T 1 / 2 was 9.17 hours.

[0274] The geometric means (synergistic CV%) of the relative bioavailability of TQS-168 after administration of 180 mg HME (regimen E) based on Cmax, AUC(0-last) and AUC(0-inf) were 56.6% (33.9%), 58.9% (30.6) and 59.6% (30.0%) compared to administration of 180 mg SDD (regimen D). The geometric means (synergistic CV%) of the relative bioavailability of the metabolite TQS-621 after administration of 180 mg TQS-168HME based on Cmax, AUC(0-last) and AUC(0-inf) were 64.9% (16.4%), 63.5% (13.8) and 63.6% (13.9%) compared to administration of 180 mg SDD. The geometric mean Cmax, AUC(0-last) and AUC(0-inf) of TQS-168 following administration of 180 mg HME (regimen E) resulted in a 2.32-, 2.14- and 2.06-fold increase, respectively, when compared with its MC counterpart (regimen B).

[0275] The geometric mean Cmax, AUC(0-last) and AUC(0-inf) of TQS-621 following administration of 180 mg HME resulted in a 2.42-, 2.46- and 2.44-fold increase, respectively, compared to its MC counterpart.

[0276] In summary, the SDD formulation showed significant improvement in exposure over the MC and HME formulations at the same TQS-168 dosage, as well as in exposure of the metabolite TQS-621. See Figures 48A-B.

[0277] 5.17. (Example 17) Part 2: Multiple-Dose Phase 1 Study - A double-blind, randomized study of subjects receiving multiple doses of TQS-168 or placebo 5.17.1. Study Design Part 2 was a double-blind, randomized, placebo-controlled clinical trial conducted to characterize and compare the pharmacokinetic (PK) profiles of TQS-168 and its metabolite TQS-621 following multiple doses of TQS-168 spray-dried dispersion (SDD) powder for oral suspension formulation in healthy subjects. See Table 32 for description of dosing regimens. [Table 32]

[0278] This randomized, double-blind, placebo-controlled, Phase 1, multiple-dose study was conducted in subjects with a body mass index (BMI) of 18.0 to 32.0 kg / m2 as measured at screening. 2 The study was conducted on healthy male subjects aged 18-55 years with pulmonary embolism. All subjects weighed at least 55 kg at screening. Key criteria for exclusion were subjects with current evidence of SARS-CoV-2 infection, clinical manifestations of significant cardiovascular, renal, hepatic, skin, chronic respiratory or gastrointestinal disease, or aspartate aminotransferase (AST) or alanine aminotransferase (ALT) >1.5 times the upper limit of normal (ULN). Subjects were recruited at a single site in the UK. Each patient provided written informed consent.

[0279] The study was conducted in three cohorts. All subjects were admitted the morning before dosing (day -1) and stayed in the facility until 48 hours after the last dose (day 9). The screening period was 4 weeks. After confirming eligibility, subjects were randomly assigned to receive either IMP (TQS-168) or placebo treatment. Subjects received IMP or placebo on the mornings of days 1-7 (approximately 24-hour intervals). Dosing was performed in the fasted state (regimen I) after an overnight fast (minimum 10 hours) or in the fed state (after a standard pre-dose or high-fat meal given 30 minutes before dosing). Safety was assessed continuously throughout the study by monitoring adverse events and use of concomitant medications, electrocardiograms (ECGs), vital signs, laboratory safety assessments, and physical examinations. Blood samples for pharmacokinetic assessments were collected from each subject from Day -1, prior to each dose (within 1 hour), and at intervals throughout the study, up to 48 hours after the final dose, if applicable.

[0280] Pharmacokinetic Evaluation: Blood samples for plasma PK analysis were collected at regular time intervals. Venous blood samples were collected from subjects by trained members of the clinical team. Pre-dose samples were collected within 1 hour prior to dosing. Timestamp 0-1 hour post-dose samples were collected within ±2 minutes of the baseline post-dose sampling time. Timestamp 1.5-12 hour post-dose samples were collected within ±10 minutes of the baseline post-dose sampling time. Timestamp 16-48 hour post-dose samples were collected within ±30 minutes of the baseline post-dose sampling time. Samples were collected in appropriate containers and processed to isolate plasma. PK analysis was performed on plasma samples using validated biochemical analytical methods.

[0281] Statistical Analysis: The sample size for the study was selected based on practical considerations and experience from previous studies of similar design. The number of subjects in each cohort was deemed adequate to evaluate the primary objective of each study. Pharmacokinetic parameters were determined by non-compartmental techniques using WinNonlin software version 8.0 or higher (Certara USA. Inc., USA). All data were enumerated and summarized by subject group using descriptive statistics. All statistical analyses were performed using SAS version 9.4 or higher.

[0282] 5.17.2. Spray-dried dispersion (SDD) formulations and oral suspensions 5.17.2.1 Preparation of SDD formulations of 2-(4-tert-butylphenyl)-1H-benzimidazole (compound of formula I) Spray dried dispersions (SDD) of 2-(4-tert-butylphenyl)-1H-benzimidazole (compound of formula I) having the compositions shown in Table 20 were prepared by spray drying the feedstock formulations shown in Table 21. [Table 20-2] [Table 21-2]

[0283] 5.17.2.2 Manufacturing Procedure for Spray-Dried Feedstock Blends The compound of formula I (45.0 g) was slowly added to 2-propanol (1791.1 g) with stirring, placed under a homogenizer (Silverson SL2 homogenizer) and stirred for 5 minutes or more until the compound of formula I was completely dissolved. The reaction mixture was then removed from the homogenizer, polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (Soluplus) (60.0 g) was slowly added with stirring, placed back in the homogenizer and stirred for 10 minutes or more until the Soluplus was completely dissolved. The reaction mixture was then removed from the homogenizer, amorphous silicon dioxide (Syloid® 244 FP) was slowly added with stirring, placed back in the homogenizer and stirred for an additional 15 minutes or more until the amorphous silicon dioxide was completely dispersed. The resulting suspension is referred to herein as the "feedstock blend."

[0284] 5.17.2.3 Manufacturing Procedures for Spray Dried Dispersion (SDD) Formulations of Compounds of Formula I A spray dryer unit (ProCepT 4M8 Spray Dryer) was equipped with a compressed air supply. Once the outlet temperature had stabilized, the feed pump was started and 2-propanol (blank solution) was sprayed as a fine spray through the nozzle into the collection chamber. The spray dryer parameters were adjusted to achieve a feed rate within the ranges shown in Table 22 below.

[0285] The feed blend was stirred under the homogenizer at an appropriate speed to maintain a uniform dispersion without creating air bubbles.The feed blend was then sprayed as a fine spray through the nozzle into the collection chamber of the spray dryer unit (ProCepT 4M8 SprayDryer, prepared with blank solution and using parameters as outlined in Table 3), where the solvent quickly evaporated to produce particles (SDD formulation of the compound of formula I) containing the compound of formula I, polyvinylcaprolactam-polyvinylacetate-polyethyleneglycol graft copolymer (Soluplus) and silicon dioxide (Syloid® 244 FP).Once all the feed blend was sprayed and collected, the feed blend was replaced with 2-propanol (blank solution) and sprayed through the nozzle of the spray dryer for 5 minutes or more to collect any remaining "feed blend" in the air stream. [Table 22-2] 5.17.2.1 Oral suspension of a spray-dried dispersion (SDD) formulation of 2-(4-tert-butylphenyl)-1H-benzimidazole (compound of formula I) A spray dried dispersion (SDD) of 2-(4-tert-butylphenyl)-1H-benzimidazole (compound of formula I) (e.g., 60-1000 mg) having the composition shown in Table 20 was reconstituted as an oral suspension in 100 g of vehicle consisting of PEG 300 (10 g), glycerol monocaprylocaprate (Capmul MCM, 0.40 mg) in sterile water for irrigation (qs to 100 g).

[0286] Regimen I: Subjects were given an oral dose of 120 mg of TQS-168 spray-dried dispersion (SDD) powder in oral suspension or placebo once daily for 7 consecutive days in the fasted state.

[0287] Regimen J: Subjects received an oral dose of 90 mg of TQS-168 spray-dried dispersion (SDD) powder in oral suspension or placebo once daily for 7 consecutive days in the fed state. Subjects were given a high-fat breakfast on days 1 and 7 and a standard breakfast on days 2-6.

[0288] Regimen K: Subjects received an oral dose of 300 mg of TQS-168 spray-dried dispersion (SDD) powder in oral suspension or placebo once daily for 7 consecutive days in the fed state. Subjects were given a high-fat breakfast on days 1 and 7 and a standard breakfast on days 2-6.

[0289] 5.17.3. Results Healthy male subjects received multiple oral doses of either 120 mg TQS-168 spray-dried dispersion (SDD) powder in the fasted state (Regimen I) or 90 mg TQS-168 SDD in the fed state (Regimen J). Plasma concentrations of TQS-168 and metabolite TQS-621 were measured over time, and key pharmacokinetic parameters were determined.

[0290] Table 33 presents the geometric means of important pharmacokinetic parameters of TQS-168 and metabolite TQS-621 in subjects following oral administration of TQS-168. [Table 33-1] [Table 33-2]

[0291] TQS-168 Regimen I PK Profile: Subjects in Cohort 1 received a single dose of TQS-168 SDD 120 mg orally QD in the fasted state for 7 consecutive days.

[0292] On Day 1, after a single dose of Regimen I, plasma concentrations of TQS-168 were measurable in all subjects beginning 0.5 hours post-dose and remained measurable between 16 and 24 hours post-dose. TQS-621 concentrations on Day 1 were also measurable in all subjects beginning 0.5 hours post-dose and remained measurable up to 24 hours post-dose.

[0293] Maximum plasma TQS-168 concentrations on day 1 occurred between 0.5 and 2.0 hours post-dose with a median Tmax of 1.0 hour post-dose. Geometric mean (CV%) Cmax and AUC(0-tau) values ​​were 172 ng / mL (49.3%) and 438 ng / mL, respectively. * h / mL (55.1%). Maximum plasma TQS-621 concentrations on Day 1 occurred between 1.0 and 4.0 hours post-dose with a median Tmax of 1.5 hours post-dose. Geometric mean (CV%) Cmax and AUC(0-tau) values ​​were 331 ng / mL (40.3%) and 2270 ng / mL (40.3%), respectively. * h / mL (35.7%). See Figures 49-50.

[0294] After multiple dosing, plasma concentrations of TQS-168 and metabolite TQS-621 were quantifiable at pre-dose on day 7 in all but one subject who was quantifiable at 0.5 hours post-dose, and remained quantifiable in all subjects between 16 and 48 hours post-dose for the final sampling time point.

[0295] Maximum plasma TQS-168 concentrations on day 7 occurred between 0.5 and 1.5 hours post-dose with a median Tmax of 0.75 hours post-dose. Concentrations then declined, resulting in a mean elimination half-life of 7.7 hours. Geometric mean (CV%) Cmax and AUC(0-tau) values ​​were 273 ng / mL (105.8%) and 692 ng / mL, respectively. * h / mL (89.8%). The geometric mean (CV%) accumulation ratios were 1.59 (88.1%) and 1.58 (51.6%) based on Cmax and AUC(0-tau), respectively. See Figures 49-50.

[0296] Maximum plasma TQS-621 concentrations on day 7 occurred between 1.0 and 4.0 hours post-dose with a median Tmax of 1.75 hours post-dose. Concentrations then declined, resulting in a mean elimination half-life of 10.2 hours. Geometric mean (CV%) Cmax and AUC(0-tau) values ​​were 340 ng / mL (39.5%) and 3320 ng / mL (1.2%), respectively. * h / mL (42.1%). The geometric mean (CV%) accumulation ratios were 1.30 (38.7%) and 1.46 (30.8%) based on Cmax and AUC(0-tau), respectively. See Figures 49-50.

[0297] Regimen I PK Overview. Cohort 1 subjects received Regimen I, TQS-168 SDD 120 mg QD for 7 consecutive days in the fasted state. On Day 1, a TQS-168 Cmax of 172 ng / mL (0.69 μM) was achieved. On Day 7 of continuous dosing, a significantly higher TQS-168 Cmax of 273 ng / mL (1.09 μM) was achieved. Results are plotted in Figures 40-41. A similar increase was observed in AUC measured exposure (Day 1 AUC(0-tau)=438 hr * ng / mL, 7th day AUC(0-tau)=692hr * Metabolite TQS-621 PK profile: Cohort 1 demonstrated a Day 1 metabolite TQS-621 Cmax of 331 ng / mL (1.24 μM) and a Day 7 Cmax of 340 ng / mL (1.27 μM). Results are plotted in Figures 42-43. AUC(0-tau)=2270hr * Metabolite exposure in ng / mL was recorded on day 1. On day 7, AUC(0-tau)=3220hr * Metabolite exposure in ng / m was obtained, see Figures 49-50.

[0298] TQS-168 Regimen J PK Profile: Cohort 2 subjects received a single dose of TQS-168 SDD 90 mg orally QD under fed conditions for 7 consecutive days.

[0299] Following a single dose of 90 mg of TQS-168 spray dried dispersion (SDD) powder for oral suspension on Day 1, plasma concentrations of TQS-168 were measurable beginning 0.5 hours post-dose in all subjects and remained measurable between 16 and 24 hours post-dose. TQS-621 concentrations on Day 1 were also measurable beginning 0.5 hours post-dose in all subjects and remained measurable until 24 hours post-dose.

[0300] Maximum plasma TQS-168 concentrations on Day 1 occurred between 1.5 and 4.0 hours post-dose with a median Tmax of 3.0 hours post-dose. Geometric mean (CV%) Cmax and AUC(0-tau) values ​​were 47.4 ng / mL (27.4%) and 223 ng.h / mL (30.9%), respectively. Comparison with Regimen I is illustrated in Figures 40A-B.

[0301] Maximum plasma TQS-621 concentrations on Day 1 occurred between 4.0 and 6.0 hours post-dose with a median Tmax of 4.0 hours post-dose. Geometric mean (CV%) Cmax and AUC(0-tau) values ​​were 189 ng / mL (40.1%) and 1400 ng / mL (10.2%), respectively. * h / mL (42.6%). Comparison with Regimen I is shown in Figures 42A-B.

[0302] On Day 1, no individual subjects exceeded the maximum tolerated Cmax or maximum tolerated AUC(0-24) (based on AUC(0-tau), tau = 24 hours) values. The maximum individual TQS-168 Cmax on Day 1 was 65.8 ng / mL, which was 21.6% of the Cmax exposure limit. The maximum individual TQS-168 AUC(0-tau) was 299 ng / mL. * h / mL, which was 10.9% of the AUC(0-24) exposure limit.

[0303] Following multiple doses of 90 mg of TQS-168 spray-dried dispersion (SDD) powder for oral suspension to healthy subjects in a fed state for 7 days, plasma concentrations of TQS-168 were quantifiable at pre-dose and remained quantifiable between 16 and 36 hours post-dose in all but three subjects who were quantifiable at 0.5 hours post-dose. TQS-621 concentrations were quantifiable at pre-dose in all subjects and remained quantifiable until the final sampling time point 48 hours post-dose.

[0304] Maximum plasma TQS-168 concentrations on Day 7 occurred between 1.0 and 4.0 hours post-dose with a median Tmax of 3.0 hours post-dose and a geometric mean elimination half-life of 4.46 hours. Geometric mean (CV%) Cmax and AUC(0-tau) values ​​were 48.8 ng / mL (48.3%) and 227 ng.h / mL (46.7%), respectively. Geometric mean (CV%) accumulation ratios were 1.03 (37.5%) and 1.24 (22.8%) based on Cmax and AUC(0-tau), respectively. Comparison with Regimen I is illustrated in Figures 41A-B.

[0305] Maximum plasma TQS-621 concentrations on Day 7 occurred between 4.0 and 6.0 hours post-dose with a median Tmax of 5.0 hours post-dose. Concentrations then declined generally biphasically resulting in a geometric mean elimination half-life of 8.65 hours. Geometric mean (CV%) Cmax and AUC(0-tau) values ​​were 218 ng / mL (37.6%) and 2060 ng.h / mL (54.8%), respectively. Geometric mean (CV%) accumulation ratios were 1.15 (12.4%) and 1.47 (18.0%) based on Cmax and AUC(0-tau), respectively. Comparison with Regimen I is illustrated in Figures 43A-B.

[0306] On Day 7, no individual subjects exceeded the maximum tolerated Cmax or maximum tolerated AUC(0-24) (based on AUC(0-tau), tau=24 hours) values ​​for TQS-168. The maximum individual TQS-168 Cmax on Day 7 was 94.1 ng / mL, which was 30.9% of the Cmax exposure limit. The maximum individual TQS-168 AUC(0-tau) was 535 ng.h / mL, which was 19.5% of the AUC(0-24) exposure limit.

[0307] Regimen J PK Overview. Cohort 2 subjects received Regimen J, given a once daily dose of TQS-168 SDD 90 mg in a fed state, resulting in a Day 1 TQS-168 Cmax of 47.4 ng / mL (0.19 μM). On Day 7 of continuous dosing of TQS-168 SDD 90 mg in a fed state, a TQS-168 Cmax of 48.8 ng / mL (0.19 μM) was observed. Results are plotted in Figures 40-41. A similar picture was observed in the comparison of exposure profiles (Day 1 AUC(0-tau)=223 hr * ng / mL, 7th day AUC(0-tau)=227hr * ng / mL. Regarding the TQS-621 PK profile, the cohort demonstrated a day 1 metabolite TQS-621 Cmax of 189 ng / mL (0.71 μM) and a day 7 Cmax of 218 ng / mL (0.82 μM). The results are plotted in Figures 42-43. AUC(0-tau)=1400hr * Metabolite exposure in ng / mL was recorded on day 1. On day 7, AUC(0-tau)=2060hr * This resulted in metabolite exposures of ng / m.

[0308] TQS-168 Regimen K PK Profile: Subjects in Cohort 3 received a single dose of TQS-168 SDD 300 mg QD under fed conditions for 7 consecutive days.

[0309] Following a single dose of 300 mg of TQS-168 spray dried dispersion (SDD) powder for oral suspension on Day 1, plasma concentrations of TQS-168 were quantifiable from 0.5 hours post-dose in all subjects and remained quantifiable in all subjects until the final sampling time point of 24 hours post-dose.

[0310] Maximum plasma TQS-168 concentrations on Day 1 occurred between 1.5 and 4.0 hours post-dose with a median Tmax of 2.0 hours post-dose. Geometric mean (CV%) Cmax and AUC(0-tau) values ​​were 229 ng / mL (38.3%) and 1210 ng / mL (12.2%), respectively. * h / mL (55.1). For a graphical comparison with Regimen I and J, see Figures 51A-B.

[0311] Maximum plasma TQS-621 concentrations on Day 1 occurred between 4.0 and 10.0 hours post-dose with a median Tmax of 4.0 hours post-dose. Geometric mean (CV%) Cmax and AUC(0-tau) values ​​were 1000 ng / mL (24.6%) and 9730 ng / mL, respectively. * h / mL (39.9%). For a graphical comparison with Regimen I and J, see Figures 53A-B.

[0312] After multiple dosing, day 7 plasma concentrations of TQS-168 and metabolite TQS-621 were quantifiable at pre-dose in all subjects and remained quantifiable between 24 and 48 hours post-dose. Concentrations of TQS were also quantifiable at the pre-dose time point in all subjects and remained quantifiable until the final sampling time point 48 hours post-dose.

[0313] Maximum plasma TQS-168 concentrations on day 7 occurred between 0.50 and 4.0 hours post-dose with a median Tmax of 4.0 hours post-dose. Concentrations then declined, resulting in a mean elimination half-life of 5.67 hours. Geometric mean (CV%) Cmax and AUC(0-tau) values ​​were 400 ng / mL (79.3%) and 2010 ng / mL (10.1%), respectively. *h / mL (88.3%). The geometric mean (CV%) accumulation ratios were 1.74 (54.6%) and 1.66 (29.5%) based on Cmax and AUC(0-tau), respectively. For a graphical comparison with regimens I and J, see Figures 52A-B.

[0314] Maximum plasma TQS-621 concentrations on day 7 occurred between 4.0 and 6.0 hours post-dose with a median Tmax of 5.0 hours post-dose. Concentrations then declined, resulting in a mean elimination half-life of 7.29 hours. Geometric mean (CV%) Cmax and AUC(0-tau) values ​​were 1300 ng / mL (36.7%) and 14,900 ng / mL (36.7%), respectively. * h / mL (61.2%). The geometric mean (CV%) accumulation ratios were 1.30 (32.9%) and 1.53 (26.1%) based on Cmax and AUC(0-tau), respectively. For graphical comparison with regimens I and J, see Figures 54A-B.

[0315] Regimen K PK Summary: Cohort 1 subjects received Regimen K, TQS-168 SDD 300 mg QD for 7 consecutive days in a fed state. On day 1, a TQS-168 Cmax of 229 ng / mL (0.91 μM) was achieved. On day 7 of continuous dosing, a significantly higher TQS-168 Cmax of 400 ng / mL (1.60 μM) was achieved. Results are plotted in Figures 40-41. A similar increase was observed with AUC measured exposure (Day 1 AUC(0-tau)=1210 hr * ng / mL, 7th day AUC(0-tau)=2010hr * Cohort 1 showed a day 1 metabolite TQS-621 Cmax of 1000 ng / mL (3.76 μM) and a day 7 Cmax of 1300 ng / mL (4.88 μM). The results are plotted in Figures 42-43. AUC(0-tau)=9730hr * Metabolite exposure in ng / mL was recorded on day 1. On day 7, AUC(0-tau)=14900hr * Metabolite exposures in ng / m were obtained.

[0316] In summary, part 2 of this study administered seven consecutive QD doses of TQS-168 in fed or fasted states to healthy male subjects at different dosages of TQS-168 SDD powder for oral suspension. Data reveal that increasing doses correspond with increasing plasma concentrations of TQS-168 and the TQS-168 metabolite TQS-621.

[0317] 6. Equivalents and Incorporation by Reference While the present invention has been shown and described in detail with reference to preferred and various alternative embodiments, it will be understood by those skilled in the relevant art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.

[0318] All references, issued patents, and patent applications cited within the body of this specification are hereby incorporated by reference in their entirety for all purposes.

Claims

1. A pharmaceutical composition for reducing neuroinflammation and / or treating neurodegenerative diseases in a human subject, wherein the pharmaceutical composition comprises a compound of formula (I) 【Chemical Formula 14】 Comprising (TQS-168) or a pharmaceutically acceptable salt thereof, the pharmaceutical composition is administered to a subject having neuroinflammation and / or neurodegenerative disease at at least one dose of the pharmaceutical composition, and after administration, the average peak concentration of TQS-168 in plasma is at least 750 ng / mL (C ｍａｘ ) and / or an amount and / or dosage that results in an oral administration, characterized in that it is a pharmaceutical composition.

2. The pharmaceutical composition according to claim 1, characterized in that the pharmaceutical composition is administered in an amount that results in an average plasma Cmax of at least 1000 ng / mL of TQS-168 after administration.

3. The pharmaceutical composition according to claim 2, characterized in that the pharmaceutical composition is administered in an amount that results in an average plasma Cmax of at least 1250 ng / mL of TQS-168 after administration.

4. The pharmaceutical composition according to claim 3, characterized in that the pharmaceutical composition is administered in an amount that results in an average plasma Cmax of at least 1500 ng / mL of TQS-168 after administration.

5. The pharmaceutical composition according to claim 4, characterized in that the pharmaceutical composition is administered in an amount that results in an average plasma Cmax of at least 1750 ng / mL of TQS-168 after administration. **Claim 6**: The pharmaceutical composition according to any one of claims 1 to 5, wherein the pharmaceutical composition is administered in an amount that provides at least 3000 ng·hr / ml of TQS-168 AUC after administration. ０－ｔ ​

7. The pharmaceutical composition according to claim 6, wherein the pharmaceutical composition is administered in an amount that provides at least 4000 ng·hr / ml of TQS-168 AUC after administration. ０－ｔ ​

8. The pharmaceutical composition according to claim 7, wherein the pharmaceutical composition is administered in an amount that provides at least 5000 ng·hr / ml of TQS-168 AUC after administration. ０－ｔ ​

9. The pharmaceutical composition is administered in an amount that provides at least 5500 ng·hr / ml of TQS-168 AUC after administration ０－ｔ The pharmaceutical composition according to claim 8, characterized in that it is administered in an amount that brings about

10. The pharmaceutical composition is administered in an amount that provides at least 6000 ng·hr / ml of TQS-168 AUC after administration ０－ｔ The pharmaceutical composition according to claim 9, characterized in that it is administered in an amount that brings about the same.

11. The pharmaceutical composition is administered in an amount that provides a TQS-168 AUC of about 7000 ng·hr / ml after administration ０－ｔ The pharmaceutical composition according to claim 10, characterized in that it is administered in an amount that provides the same

12. The pharmaceutical composition according to claim 1, wherein the time (Tmax) to the plasma Cmax of TQS-168 is 2 hours or less.

13. The pharmaceutical composition according to claim 12, wherein the Tmax is 90 minutes or less.

14. The pharmaceutical composition according to claim 13, wherein the Tmax is 75 minutes or less.

15. The pharmaceutical composition according to claim 14, wherein the Tmax is about 60 minutes.

16. A pharmaceutical composition for reducing neuroinflammation and / or treating neurodegenerative diseases in a human subject, wherein the pharmaceutical composition comprises a compound of formula (I) 【Chemical Formula 15】 (TQS-168), or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition comprises at least 1000 ng / mL of the compound of formula (II) in a subject having neuroinflammation and / or neurodegenerative diseases after administration 【Chemical 16】 The pharmaceutical composition is characterized by being orally administered in an amount that provides an average peak plasma concentration (C ｍａｘ ) of (TQS-621).

17. The pharmaceutical composition is administered in an amount that results in a plasma C of TQS-621 of 200 to 2750 ng / mL after administration ｍａｘ The pharmaceutical composition according to claim 16, characterized in that it is administered in an amount that brings about

18. The pharmaceutical composition according to claim 17, characterized in that the pharmaceutical composition is administered in an amount that results in a plasma Cmax of 300-2200 ng / mL of TQS-621 after administration.

19. The pharmaceutical composition according to claim 18, characterized in that the pharmaceutical composition is administered in an amount that results in a plasma Cmax of 400-1800 ng / mL of TQS-621 after administration.

20. A pharmaceutical composition for reducing neuroinflammation and / or treating neurodegenerative diseases in a human subject, wherein the pharmaceutical composition comprises a compound of formula (I) 【Chemical 17】 (TQS-168), or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is administered orally in an amount that, after administration, results in (a) The average peak concentration (C ｍａｘ ) of TQS-168 in plasma of at least 750 ng / mL (b) The average time (T ｍａｘ up to C of TQS-168 in plasma below 75 minutes ｍａｘ ) together with; and (c) a compound of formula (II) in plasma of at least 1000 ng / mL 【Chemical Formula 18】 The average peak concentration (C ｍａｘ ) of (TQS-621) was (d) Average time (T ｍａｘ to C of TQS-621 in plasma for 4 hours or less ｍａｘ ) in a subject having neuroinflammation and / or neurodegenerative disease. A pharmaceutical composition, characterized in that it is administered orally in an amount that results in

21. The pharmaceutical composition according to any one of Claims 1, 16, and 20, characterized in that the pharmaceutical composition is administered at a daily oral dose of 200 to 800 mg of TQS-168 or a salt thereof.

22. The pharmaceutical composition according to Claim 21, characterized in that the pharmaceutical composition is administered at a daily oral dose of 300 to 700 mg of TQS-168 or a salt thereof.

23. The pharmaceutical composition according to Claim 22, characterized in that the pharmaceutical composition is administered at a daily oral dose of 400 to 600 mg of TQS-168 or a salt thereof.

24. The pharmaceutical composition according to Claim 23, characterized in that the pharmaceutical composition is administered at a daily oral dose of 400 to 500 mg of TQS-168 or a salt thereof.

25. The pharmaceutical composition according to Claim 24, characterized in that the pharmaceutical composition is administered at a daily oral dose of 400 mg or 450 mg of TQS-168 or a salt thereof.

26. The pharmaceutical composition according to any one of Claims 1, 16, and 20, characterized in that the pharmaceutical composition is a liquid suspension.

27. The pharmaceutical composition according to any one of Claims 1, 16, and 20, characterized in that the pharmaceutical composition is a liquid solution.

28. The pharmaceutical composition according to any one of Claims 1, 16, and 20, characterized in that the pharmaceutical composition is in a solid dosage form.

29. The pharmaceutical composition according to Claim 28, wherein TQS-168 or a salt thereof is crystalline.

30. The pharmaceutical composition according to Claim 28, wherein TQS-168 or a salt thereof is amorphous.

31. The pharmaceutical composition according to Claim 30, characterized in that the pharmaceutical composition is in the form of a spray-dried dispersion.

32. The pharmaceutical composition according to Claim 30, characterized in that the pharmaceutical composition is in the form of a hot-melt extrudate.

33. The pharmaceutical composition according to Claim 28, wherein the solid dosage form is a sachet.

34. The pharmaceutical composition according to Claim 28, wherein the solid dosage form is a capsule.

35. The pharmaceutical composition according to claim 28, wherein the solid dosage form is a tablet.

36. The pharmaceutical composition according to any one of claims 1, 16, and 20, wherein the subject has a neurodegenerative disease selected from motor neuron disease, amyotrophic lateral sclerosis (ALS), Alzheimer's disease, vascular dementia, frontotemporal lobar degeneration (frontotemporal dementia), Lewy body dementia, Parkinson's disease, Huntington's disease, demyelinating diseases, and multiple sclerosis (MS).

37. The pharmaceutical composition according to claim 36, wherein the subject has motor neuron disease.

38. The pharmaceutical composition according to claim 37, wherein the subject has ALS.

39. The pharmaceutical composition according to claim 36, wherein the subject has Alzheimer's disease.

40. The pharmaceutical composition according to any one of claims 1, 16, and 20, wherein the subject is at least 40 years old and does not have a previously diagnosed neurodegenerative disease.

41. The pharmaceutical composition according to claim 40, wherein the subject is at least 60 years old.

42. The pharmaceutical composition according to claim 41, wherein the subject is at least 65 years old.

Images