Inhibitory serotonin transporter protein (5-HTT)

Abstract

Disclosed are compositions containing mesembranol, such as (-) mesembranol or (-)6-epi-mesembrananol. The disclosed compositions can be used in methods for inhibiting SERT. Also provided are methods for treating anxiety and depression, including methods comprising orally administering a therapeutically effective amount of a particular pharmaceutical composition to a subject in need thereof.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 434,726, filed December 22, 2022, the contents of which are incorporated herein by reference in their entirety. [Technical Field]

[0002] The present disclosure relates to the pharmaceutical field, including the discovery of alkaloid compounds useful for inhibiting the serotonin transporter protein (5-HTT). [Background technology]

[0003] Plants in the Sceletium genus contain indole alkaloids with biological activity useful in the treatment of mental health conditions, such as mild to moderate depression. Natural extracts of Sceletium tortuosum, an indigenous herb from South Africa also known as "kougoed," "channa," or "kanna," may contain pharmacologically active alkaloids. Among the alkaloids listed below, mesembrine, mesembrenone, and mesembrenol are present in Sceletium tortuosum extracts used to treat anxiety, stress, and mental health conditions. [ka]

[0004] Mesembranol has not been well characterized pharmacologically, and its concentration in plant extracts is relatively low compared to other bioactive alkaloids of Sceletium tortuosum. Mesembranol is present in little or no extracts from Sceletium tortuosum plants. Figures 1A–1C show the liquid chromatography–mass spectrometry (LC-MS) chemical fingerprints of alkaloids in Sceletium tortuosum plants collected from three different regions of South Africa where Sceletium tortuosum grows (Non-Patent Document 1). In each chromatogram, the peak at 3.77 min is attributed to mesembranol. The presence of mesembranol was detected only in trace amounts in samples from Northern Cape (Figure 1A) and Western Cape 1 (Figure 1B). Mesembranol was not a major alkaloid, and its abundance in Western Cape 2 samples was moderate (it was the fourth most abundant alkaloid in Western Cape 2 samples; Figure 1C). These data indicate that extraction of the Sceletium tortuosum plant is unlikely to yield mesembranol in sufficient quantities to provide a therapeutic effect.

[0005] Additionally, extracts derived from Sceletium tortuosum contain numerous biologically active alkaloids that confer various pharmacological effects. For example, reported pharmacological activities of Sceletium tortuosum extracts include serotonin transporter inhibition (5-HTT), phosphodiesterase-4 inhibition (PDE4), monoamine oxidase A (MAO-A) inhibitory activity, acetylcholinesterase (AChE) inhibitory activity, GABA receptor binding activity, opioid receptor binding activity, dopamine transporter inhibition, AMPA receptor modulation, and effects on vesicular monoamine transporter-2 (VMAT-2) and subsequent monoamine release.

[0006] Analysis of a standardized commercially available extract of Sceletium tortuosum (obtained under the trade name Zembrin®) was reported in 2011 to have 0.35% to 0.45% total alkaloids, with mesembrenone and mesembrenol accounting for more than 60%, with mesembrine contributing less than 20% (see Non-Patent Documents 2 and 3). The extract demonstrated greater than 80% inhibition of the serotonin (5-HT) transporter, demonstrating the potency of isolated alkaloids at the 5-HT transporter as shown in Table A below (Non-Patent Document 2). Referring to the data in Table A, concentration-dependent inhibition was observed, with mesembrine being the more active compound in the 5-HT transporter assay (i.e., 20-fold more potent than mesembrenone and 87-fold more potent than mesembrenol). A toxicological safety evaluation of this standardized extract was subsequently reported in 2014 (Non-Patent Document 3). TIFF2025542199000002.tif55165

[0007] However, bioactive plant extracts for therapeutic consumption can vary widely both seasonally and among different Sceletium tortuosum plants, making it impossible to provide a sufficiently reproducible and stable phytochemical profile of the desired bioactive components. Sceletium plants and their extracts can vary significantly in total alkaloid content, as well as the chemical nature and relative concentrations of alkaloids from individual Sceletium plants. Furthermore, mesembranol concentrations in Sceletium tortuosum can vary across different regions of South Africa, and mesembranol content has been reported to be relatively low in most plant extracts tested. Finally, Sceletium alkaloids can be unstable under various conditions that may occur during extraction from plant materials and during storage and formulation of the extracts.

[0008] In Sceletium tortuosum extracts, mesembranol and its isomer 6-epi-mesembrananol are present in low concentrations compared to other major alkaloids. Therefore, mesembranol and 6-epi-mesembrananol have not been well characterized in the scientific literature. The pharmacological activity and selectivity of mesembranol or 6-epi-mesembrananol have not been reported. Their therapeutic use has been limited by the low content, variability, and instability of these alkaloids in natural extracts, as well as the instability and pharmacokinetic profiles of these compounds obtained from natural products. Naturally occurring mesembranol and 6-epi-mesembrananol are serotonin transporter inhibitors with high specificity compared to other aforementioned pharmacological targets. Sceletium tortuosum alkaloid extracts do not produce sufficient levels of mesembranol and 6-epi-mesembrananol compared to other components, and the extracts may also exert other pharmacological effects that are undesirable for the treatment of anxiety or depression.

[0009] There remains an unmet need for pharmaceutical compositions containing highly pure, predictable, stable, and reproducible forms of therapeutic alkaloid compounds, such as mesembranol and 6-epi-mesembrananol. Additionally, there is a need for oral pharmaceutical compositions that provide pure therapeutic alkaloid compositions with desirable pharmacokinetic properties upon administration. Finally, there is an unmet need for pharmaceutical compositions that contain properties significantly different from naturally derived compositions obtained from plant extracts. [Prior art documents] [Non-patent literature]

[0010] [Non-Patent Document 1] J. Zhao et al., Phytochemistry, 2018 [Non-patent document 2] Harvey et al., “Pharmacological actions of the South African medicinal and functional food plant Sceletium tortuosum and its principal alkaloids,” Journal of Ethnopharmacology 137 (2011) 1124-1129 [Non-patent document 3] Murbach et. al., “A toxicological safety assessment of a standardized extract of Sceletium tortuosum (Zembrin®) in rats,” Food and Chemical Toxicology 74 (2014) 190-199 Summary of the Invention [Means for solving the problem]

[0011] Applicants have discovered new and useful methods of using mesembranol (i.e., (-) mesembranol or (-)6-epi-mesembrananol). In some embodiments, mesembranol (Compound 18) or (-)6-epi-mesembrananol (Compound 19) can be used to selectively inhibit the serotonin transporter (5-HTT) while avoiding inhibition of phosphodiesterase-4 (PDE4) at levels at or near those observed with other pharmacologically active alkaloids identified in Sceletium tortuosum extracts. [ka]

[0012] In some embodiments, methods of using (-) mesembranol or (-)6-epi-mesembranol are based, in part, on the discovery that (-) mesembranol or (-)6-epi-mesembranol can be used to provide a mechanism of therapeutic action similar to selective serotonin reuptake inhibitors (SSRIs).

[0013] In some embodiments, methods of using (-)6-epi-mesembranol are based in part on the discovery of different pharmacokinetic data for (-)6-epi-mesembranol (Compound 019) and (-)mesembranol (Compound 018), specifically, the half-life of Compound 019 is about three times longer than that of Compound 018 and about two times longer than that of mesembrine (Compound 001) (measured after intravenous administration to dogs according to Example 7).

[0014] In some embodiments, (-)6-epi-mesembranol (compound 19) can be used to selectively inhibit the serotonin transporter (5-HTT) while avoiding inhibition of phosphodiesterase-4 (PDE4) at or near levels observed for other pharmacologically active alkaloids identified in Sceletium tortuosum extracts. In some embodiments, (-)6-epi-mesembranol (compound 19) can be used to treat anxiety or depression. In certain embodiments, methods for treating anxiety and depression include: Compound 019, [ka] Orally administer a therapeutically effective amount of a pharmaceutical composition comprising the compound or its pharmaceutically acceptable salt. In certain embodiments, the method is a method for treating major depressive disorder (MDD). In certain embodiments, the method is a method for treating generalized anxiety disorder (GAD).

[0015] In certain embodiments, Compound 019 or a pharmaceutically acceptable salt thereof is administered as a capsule or tablet. In certain embodiments, Compound 019 or a pharmaceutically acceptable salt thereof is administered once or twice daily. In certain embodiments, the pharmaceutical composition contains about 1% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0016] In certain embodiments, the method of inhibiting SERT comprises administering to a mammal in need thereof a therapeutically effective amount of [ka] or a pharmaceutically acceptable salt thereof.

[0017] In certain embodiments, the pharmaceutical composition [ka] or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition contains about 3% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof, and Compound 019 is at least 90% of the total alkaloid content in the pharmaceutical composition. In certain embodiments, the pharmaceutical composition has a PDE4 / SERT inhibition ratio of at least 10:1. In certain embodiments, the pharmaceutical composition has a half-life of Compound 019 of at least 30 minutes in a human hepatocyte assay. In certain embodiments, the pharmaceutical composition has an IC 50 is less than about 30 nM. In certain embodiments, the pharmaceutical composition has an inhibition rate of at least one of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 of less than about 6%. In certain embodiments, the pharmaceutical composition has a PDE4 / SERT inhibition ratio of at least 10:1.

[0018] In certain embodiments, a method of inhibiting SERT without inhibiting PDE4 comprises administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising Compound 019 or a pharmaceutically acceptable salt thereof. In certain embodiments, a composition comprising Compound 019 or a pharmaceutically acceptable salt thereof and less than about 0.5% mesembrine or mesembrenone as measured by HPLC.

[0019] 1. A method for treating anxiety or depression, comprising administering to a subject in need thereof Compound 019, [ka] or a pharmaceutically acceptable salt thereof. In some embodiments, the method is a method for treating major depressive disorder (MDD) or generalized anxiety disorder (GAD).

[0020] In some embodiments, Compound 019 or a pharmaceutically acceptable salt thereof is administered as a capsule or tablet. In some embodiments, Compound 019 or a pharmaceutically acceptable salt thereof is administered once or twice daily. In some embodiments, the pharmaceutical composition contains about 1% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0021] In some embodiments, the method of inhibiting SERT comprises administering to a mammal in need thereof a therapeutically effective amount of [ka] or a pharmaceutically acceptable salt thereof.

[0022] In some embodiments, the pharmaceutical composition comprises: [ka] or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition contains about 3% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof, and Compound 019 is at least 90% of the total alkaloid content in the pharmaceutical composition.

[0023] In some embodiments, the pharmaceutical composition has a PDE4 / SERT inhibition ratio of at least 10:1. In some embodiments, Compound 019 has a half-life of at least 30 minutes in a human hepatocyte assay of the pharmaceutical composition. In some embodiments, the pharmaceutical composition has an IC 50 is less than about 30 nM. In some embodiments, the pharmaceutical composition has an inhibition rate of at least one of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 of less than about 6%. In some embodiments, the pharmaceutical composition has a PDE4 / SERT inhibition ratio of at least 10:1.

[0024] In some embodiments, the method for inhibiting SERT without inhibiting PDE4 comprises administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising Compound 019 or a pharmaceutically acceptable salt thereof.

[0025] In some embodiments, a composition is provided comprising Compound 019 or a pharmaceutically acceptable salt thereof, wherein the composition contains less than about 0.5% mesembrine or mesembrenone as measured by HPLC.

[0026] In some embodiments, the method for treating anxiety and depression comprises administering to a subject in need thereof Compound 018 [ka] or a pharmaceutically acceptable salt thereof.

[0027] Numerous other embodiments are further provided that may be applied to any aspect of the invention described herein. [Brief explanation of the drawings]

[0028] [Figure 1A] Chromatogram of the Northern Cape sample, reproduced from J. Zhao et al., Phytochemistry, 2018. [Figure 1B] Chromatogram of Northern Cape sample 1, reproduced from J. Zhao et al., Phytochemistry, 2018. [Figure 1C] Chromatogram of Northern Cape sample 2, reproduced from J. Zhao et al., Phytochemistry, 2018. [Figure 2A] 1 is a graph comparing the compound stability of (-) mesembrine (triangles) and (-) mesembranol (squares) in human hepatocytes. [Figure 2B] 1 is a graph comparing the compound stability of (-) mesembrine (triangles) and (-) 6-epi-mesembranol (circles) in human hepatocytes. [Figure 3] 1 is a graph showing the pharmacokinetic profiles of Compound 018 and Compound 019 evaluated in dogs, showing the plasma concentrations (nM) of Compound 018, Compound 019, and Compound 001 following IV administration of 2 mg / kg of the compounds. [Figure 4] 1 is a graph showing the plasma concentrations (nM) of Compound 018, Compound 019, and Compound 001 following PO administration of 10 mg / kg of the compounds. DETAILED DESCRIPTION OF THE INVENTION

[0029] Described herein are compositions comprising a compound selected from mesembranol or 6-epimepimesembranol, or a pharmaceutically acceptable salt thereof, and methods of using (-) mesembranol and (-) 6-epi-mesembranol.

[0030] Applicants have discovered new and useful methods of using mesembranol (i.e., (-) mesembranol or (-)6-epi-mesembrananol). In some embodiments, mesembranol (Compound 18) or (-)6-epi-mesembrananol (Compound 19) can be used to selectively inhibit the serotonin transporter (5-HTT) while avoiding inhibition of phosphodiesterase-4 (PDE4) at levels at or near those observed with other pharmacologically active alkaloids identified in Sceletium tortuosum extracts. [ka]

[0031] In some embodiments, methods of using (-) mesembranol or (-)6-epi-mesembrananol are based, in part, on the discovery that (-) mesembranol or (-)6-epi-mesembrananol can be used to provide a mechanism of therapeutic action similar to selective serotonin reuptake inhibitors (SSRIs).

[0032] In some embodiments, a method for treating a psychiatric disorder comprises administering to a mammal in need thereof an effective amount of a compound selected from (-) mesembranol and (-)6-epi-mesembrananol, or salts thereof. In certain embodiments, the compound is (-) mesembranol. In certain embodiments, the compound is (-)6-epi-mesembrananol.

[0033] In certain embodiments, the present disclosure provides a method for inhibiting SERT, comprising therapeutically administering to a mammal in need thereof a compound selected from (-) mesembranol and (-)6-epi-mesembrananol, or a salt thereof. In certain embodiments, the mesembranol is (-) mesembranol. In certain embodiments, the compound is (-) mesembranol. In certain embodiments, the compound is (-)6-epi-mesembrananol.

[0034] In certain embodiments, the present disclosure provides a method of inhibiting SERT, comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound selected from (-) mesembranol and (-)6-epi-mesembrananol, or a salt thereof. In certain embodiments, the compound is (-) mesembranol. In certain embodiments, the compound is (-)6-epi-mesembrananol.

[0035] In certain embodiments, the pharmaceutical composition comprises (-)6-epi-mesembrananol and a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises mesembranol and a pharmaceutically acceptable excipient. In certain embodiments, the present disclosure provides a method for treating a psychiatric disorder, comprising administering a compound of the present disclosure to a subject. In certain embodiments, a method for inhibiting SERT without inhibiting PDE4 comprises administering a therapeutically effective amount of a pharmaceutical composition to a mammal in need thereof.

[0036] In certain embodiments, the composition comprises mesembranol (compound 18) or a pharmaceutically acceptable salt thereof and less than about 0.5% mesembrine or mesembrenone as measured by HPLC, hi certain embodiments, the composition comprises (-)6-epi-mesembranol (compound 19) or a pharmaceutically acceptable salt thereof and less than about 0.5% mesembrine or mesembrenone as measured by HPLC.

[0037] In certain embodiments, there is provided a method for treating anxiety, the method comprising administering to a subject in need thereof a pharmaceutical composition comprising mesembranol (or a pharmaceutically acceptable salt thereof) and / or (-)6-epi-mesembrananol (or a pharmaceutically acceptable salt thereof). In certain embodiments, there is provided a method for inhibiting serotonin transporter (5-HTT) in the central nervous system of a subject, the method comprising administering to a subject in need thereof a pharmaceutical composition comprising mesembranol (or a pharmaceutically acceptable salt thereof) and / or (-)6-epi-mesembrananol (or a pharmaceutically acceptable salt thereof), wherein the pharmaceutical composition does not inhibit PDE4A1A, PDE4B2, PDE4C1, or PDE4D2 by greater than 5% at 10 micromolar.

[0038] In some embodiments, mesembranol (Compound 18) or (-)6-epi-mesembrananol (Compound 19) can be used to selectively inhibit the serotonin transporter (5-HTT) while avoiding inhibition of phosphodiesterase-4 (PDE4) at or near levels observed with other pharmacologically active alkaloids identified in Sceletium tortuosum extract. Applicant has discovered that mesembranol (Compound 18) and (-)6-epi-mesembrananol (Compound 19) are potent inhibitors of the serotonin transporter protein (5-HTT), inhibiting PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar concentrations. Compounds 18 and 19 unexpectedly demonstrated significantly lower activity against PDE4 compared with other pharmacologically active alkaloids identified in Sceletium tortuosum extract. For example, the alkaloid (+) mesembrine (Compound 2) exhibited approximately 6- to 23-fold greater PDE4 activity as measured at 10 micromolar inhibition of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 compared to Compound 18 or Compound 19. Isolated mesembrenol stereoisomers (Compounds 20, 21, 23, and 24) exhibited approximately 3- to 13-fold greater PDE4A1A activity and approximately 3- to 12-fold greater PDE4B2 activity as measured at 10 micromolar inhibition of PDE4A1A and PDE4B2 compared to Compound 18. In some embodiments, the pharmaceutical composition includes an active ingredient (API) consisting of Compound 18 (or a pharmaceutically acceptable salt thereof) and / or Compound 19 (or a pharmaceutically acceptable salt thereof), wherein the API does not contain mesembrine or mesembrenone.

[0039] In certain embodiments, the pharmaceutical composition contains (-) mesembranol and / or (-) 6-epi-mesembrananol. In certain embodiments, the pharmaceutical composition contains about 10% or less of an alkaloid selected from (-) mesembrine, mesembrenone, and mesembrenol, alone or in combination together. For example, the pharmaceutical composition may contain less than about 10% by weight, less than about 5% by weight, less than about 3% by weight, less than about 2% by weight, or less than about 1% by weight of the following alkaloid combinations: (-) mesembrine and mesembrenone, or mesembrenone and mesembrenol, or (-) mesembrine, mesembrenone, and mesembrenol. In certain embodiments, the composition is substantially free of (-) mesembrine, mesembrenone, and mesembrenol. For example, the pharmaceutical composition can contain an alkaloid and a combination of the following alkaloids at less than about 10%, less than about 5%, less than about 3%, less than about 2%, or less than about 1% by weight of the total alkaloids: (-) mesembrine and mesembrenone, or mesembrenone and mesembrenol, or (-) mesembrine, mesembrenone, and mesembrenol. In certain embodiments, the pharmaceutical composition can contain, for example, about 0.3% to about 0.5% by weight of alkaloids, with mesembrenone and mesembrenol representing less than about 60% by weight of the total alkaloid content. In certain embodiments, the pharmaceutical composition can contain, for example, about 0.3% to about 0.5% by weight of alkaloids, with mesembrenone representing less than about 20% by weight of the total alkaloid content. In certain embodiments, the pharmaceutical composition may contain, for example, about 0.3% to about 0.5% by weight of alkaloids, and (-) mesembranol and / or (-)6-epimesembranol comprise at least about 70%, at least 80%, or at least about 90% of the total amount of alkaloids in the composition. In certain embodiments, the composition contains less than about 70 micrograms of a combination of mesembrenone and mesembrenol. In certain embodiments, the composition contains less than about 23 micrograms of (-) mesembrine.

[0040] In certain embodiments, the composition contains less than about 70 micrograms of mesembrenone and mesembrenol per 11.3 mg of total alkaloid content. In certain embodiments, the composition contains less than about 23 micrograms of (-) mesembrine per 11.3 mg of total alkaloid content.

[0041] In some embodiments, a method for treating a patient suffering from a disorder comprises administering to the patient a composition comprising a compound disclosed herein for the treatment or prevention of a psychiatric disorder. In some embodiments, a method for treating a patient suffering from a disorder comprises administering to the patient a composition comprising a compound disclosed herein for the treatment or prevention of a diagnosed condition selected from anxiety, stress, and depression. In some embodiments, the method comprises administering to a patient in need thereof a therapeutically effective amount of a compound selected from (-) mesembranol and / or (-)6-epi-mesembrananol for the treatment of depression. In some embodiments, the method comprises administering to a patient in need thereof a therapeutically effective amount of a compound selected from (-) mesembranol and / or (-)6-epi-mesembrananol for the treatment of a condition selected from the group consisting of anxiety associated with depression, anxiety with depression, and mixed anxiety and depression. In some embodiments, the method comprises administering to a patient in need thereof a therapeutically effective amount of a compound selected from (-) mesembranol and / or (-)6-epi-mesembrananol for the treatment of anxiety and hysteria, or anxiety and depression.

[0042] In certain embodiments, the present disclosure provides methods for treating a psychiatric disorder, comprising administering to a subject a compound of the present disclosure (e.g., a compound selected from (-) mesembranol and / or (-) 6-epi-mesembrananol).

[0043] In certain embodiments, the present disclosure provides a method of inhibiting SERT, comprising administering to a mammal in need thereof a therapeutically effective amount of a compound selected from (-) mesembranol and (-)6-epi-mesembrananol. In certain embodiments, the compound is (-) mesembranol. In certain embodiments, the compound is (-)6-epi-mesembrananol. In certain embodiments, the compound (e.g., (-) mesembranol or (-)6-epi-mesembrananol) is in the form of a composition according to the present disclosure.

[0044] In certain embodiments, the present disclosure provides a method for inhibiting SERT, comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound selected from (-) mesembranol and (-) 6-epi-mesembrananol, or a salt thereof. In certain embodiments, the compound is (-) mesembranol. In certain embodiments, the compound is (-) 6-epi-mesembrananol. In certain embodiments, the pharmaceutical composition contains about 10% or less of an alkaloid selected from (-) mesembrine, mesembrenone, and mesembrenol, alone or in combination together. For example, the pharmaceutical composition may contain less than about 10% by weight, less than about 5% by weight, less than about 3% by weight, less than about 2% by weight, or less than about 1% by weight of the following alkaloid combinations: (-) mesembrine and mesembrenone, or mesembrenone and mesembrenol, or (-) mesembrine, mesembrenone, and mesembrenol. In certain embodiments, the composition is substantially free of (-) mesembrine, mesembrenone, and mesembrenol.

[0045] In some embodiments, a compound disclosed herein is administered to a patient in a unit dose. In some embodiments, a compound disclosed herein is prescribed to a patient in an oral unit dose, such as a capsule or tablet, one or more times daily. In some embodiments, a compound disclosed herein is administered to a patient for the treatment of a disease or condition for which mesembranol (e.g., (-) mesembranol or (-)6-epi-mesembrananol) is safe and effective. In some embodiments, a method comprises administering to a patient in need thereof a therapeutically effective amount of (-) mesembranol (e.g., (-) mesembranol or (-)6-epi-mesembrananol) for the treatment of anxiety. In some embodiments, a method comprises administering to a patient in need thereof a therapeutically effective amount of mesembranol (e.g., (-) mesembranol or (-)6-epi-mesembrananol) for the treatment of a disease selected from the group consisting of mild to moderate depression and major depressive episodes. In some embodiments, the method comprises administering to a patient in need thereof a therapeutically effective amount of mesembranol (e.g., (-) mesembranol or (-)6-epi-mesembrananol) for the treatment of a disorder selected from the group consisting of psychological and psychiatric disorders in which anxiety is present. In some embodiments, the method comprises administering to a patient in need thereof a therapeutically effective amount of (-) mesembranol (e.g., (-) mesembranol or (-)6-epi-mesembrananol) for the treatment of a major depressive episode. In some embodiments, the method comprises administering to a patient in need thereof a therapeutically effective amount of mesembranol (e.g., (-) mesembranol or (-)6-epi-mesembrananol) for the treatment of a disorder selected from the group consisting of alcohol and drug addiction, bulimia nervosa, and obsessive-compulsive disorder. In some embodiments, an amount of mesembranol (e.g., (-) mesembranol or (-)6-epi-mesembrananol) between 20 micrograms and 2 milligrams is orally administered to the patient to treat the patient in need thereof.In some embodiments, an amount of mesembranol (e.g., (-) mesembranol or (-)6-epi-mesembrananol) in an amount of 20 micrograms to 2 milligrams is orally administered to a patient to treat a patient in need thereof.

[0046] In some embodiments, the method comprises administering to a patient in need thereof a therapeutically effective amount of mesembranol (e.g., (-) mesembranol or (-)6-epi-mesembrananol) for the treatment of a disorder selected from the group consisting of acute and maintenance treatment of major depressive disorder (MDD), acute and maintenance treatment of obsessive-compulsive disorder (OCD), acute and maintenance treatment of anorexia nervosa, and acute treatment of panic disorder (with or without agoraphobia).

[0047] In some embodiments, the method comprises administering to a patient in need thereof a therapeutically effective amount of mesembranol (e.g., (-) mesembranol or (-)6-epi-mesembrananol) for the treatment of a disorder selected from the group consisting of major depressive disorder (MDD), obsessive-compulsive disorder (OCD), panic disorder (PD), social anxiety disorder (SAD), generalized anxiety disorder (GAD), and post-traumatic stress disorder (PTSD).

[0048] In some embodiments, the method comprises administering to a patient in need thereof a therapeutically effective amount of mesembranol (e.g., (-) mesembranol or (-)6-epi-mesembranol) for the treatment of a disorder selected from the group consisting of major depressive disorder (MDD), obsessive-compulsive disorder (OCD), panic disorder (PD), post-traumatic stress disorder (PTSD), social anxiety disorder (SAD), and premenstrual dysphoric disorder (PMDD).

[0049] In certain embodiments, the (-)6-epi-mesembrananol in the pharmaceutical composition has a half-life of at least 30 minutes, at least about 60 minutes, at least about 90 minutes, or at least about 120 minutes in a human hepatocyte assay performed, e.g., according to Example 4.

[0050] In certain embodiments, the AUC of (-)6-epi-mesembrananol last to (-) mesembrine is about 2 to about 2.5, for example, in an assay performed according to Example 5. In certain embodiments, the AUC of (-) mesembranol last The ratio of (-) mesembrine to (-) mesembrine is, for example, from about 4 to about 4.5 in an assay performed according to Example 5.

[0051] In certain embodiments, compound 018 has about a 3-fold longer plasma half-life, about a 9-fold higher Cmax, and about a 4-fold greater AUC(last) than mesembrine (compound 001), e.g., in an assay performed according to Example 5. In certain embodiments, compound 019 has about a 2-fold longer plasma half-life, about a 2-fold higher Cmax, and about a 2-fold greater AUC(last) than mesembrine (compound 001), e.g., in an assay performed according to Example 5.

[0052] In certain embodiments, a method for inhibiting SERT without inhibiting PDE4 comprises administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound selected from (-) mesembranol or (-)6-epi-mesembrananol. In some embodiments, the pharmaceutical composition has an IC 50(e.g., performed according to Example 3A) is less than about 30 nM or less than about 15 nM. In certain embodiments, the pharmaceutical composition has an inhibition rate (e.g., performed according to Example 3B) of less than about 6% against at least one of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2. In some embodiments, the pharmaceutical composition has an inhibition rate (e.g., performed according to Example 3B) of less than about 6% against at least two of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2. In some embodiments, the pharmaceutical composition has an inhibition rate (e.g., performed according to Example 3B) of less than about 6% against at least three of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2. In some embodiments, the pharmaceutical composition has an inhibition rate (e.g., performed according to Example 3B) of less than about 6% against each of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0053] Pharmaceutical Composition In certain embodiments, the present application relates to a pharmaceutical composition comprising an active ingredient. In certain embodiments, the pharmaceutical composition comprises a compound disclosed herein as an active ingredient (API) and a pharmaceutically acceptable carrier comprising one or more excipients. In some embodiments, the pharmaceutical composition optionally further comprises a therapeutic compound (i.e., a drug) and a pharmaceutically acceptable carrier. The pharmaceutical composition may be a medicament.

[0054] In some embodiments, pharmaceutical compositions comprise Compound 18 and / or Compound 19, or pharmaceutically acceptable salts thereof, in the absence of one or more pharmacologically active alkaloid compounds in Sceletium tortuosum extract. Pharmaceutical compositions comprising Compound 18, Compound 19, or mixtures thereof (including pharmaceutically acceptable salts of Compound 18 and Compound 19) may be used to inhibit the serotonin transporter (5-HTT) while avoiding inhibition of phosphodiesterase-4 (PDE4) at levels observed with other pharmacologically active alkaloids identified in Sceletium tortuosum extract, such as mesembrine and mesembrenone.

[0055] In some embodiments, pharmaceutical compositions may contain mesembranol (Compound 18) and / or (-)6-epi-mesembranol (Compound 19), including mixtures of Compound 18 and Compound 19, or a pharmaceutically acceptable salt thereof, and contain less than 0.5% mesembrine or mesembrenone detectable by HPLC. In some embodiments, pharmaceutical compositions may contain mesembranol (Compound 18), or a pharmaceutically acceptable salt thereof, and contain less than 0.5% mesembrine or mesembrenone detectable by HPLC. In some embodiments, pharmaceutical compositions may contain (-)6-epi-mesembranol (Compound 19), or a pharmaceutically acceptable salt thereof, and contain less than 0.5% mesembrine or mesembrenone detectable by HPLC.

[0056] In some embodiments, the pharmaceutical composition comprises an API consisting of Compound 18 (or a pharmaceutically acceptable salt thereof) and / or Compound 19 (or a pharmaceutically acceptable salt thereof) for treating symptoms of anxiety in a subject in need thereof. In some embodiments, the pharmaceutical composition comprises an API consisting of Compound 18 (or a pharmaceutically acceptable salt thereof), which is a selective serotonin reuptake inhibitor for treating symptoms of anxiety in a subject in need thereof. In some embodiments, the pharmaceutical composition comprises an API consisting of Compound 19 (or a pharmaceutically acceptable salt thereof) for treating symptoms of anxiety in a subject in need thereof. In some embodiments, the pharmaceutical composition comprises an API consisting of Compound 18 (or a pharmaceutically acceptable salt thereof) and / or Compound 19 (or a pharmaceutically acceptable salt thereof) for treating symptoms of anxiety in a subject in need thereof, in the absence of mesembrine or mesembrenone.

[0057] In some embodiments, the pharmaceutical composition comprises an API consisting of compound 18 (or a pharmaceutically acceptable salt thereof) and / or compound 19 (or a pharmaceutically acceptable salt thereof) for treating generalized anxiety disorder (GAD) in a subject in need thereof. In some embodiments, the pharmaceutical composition comprises an API consisting of compound 18 (or a pharmaceutically acceptable salt thereof) for treating generalized anxiety disorder (GAD). In some embodiments, the pharmaceutical composition comprises an API consisting of compound 19 (or a pharmaceutically acceptable salt thereof) for treating generalized anxiety disorder (GAD). In some embodiments, the pharmaceutical composition comprises an API consisting of compound 18 (or a pharmaceutically acceptable salt thereof) and / or compound 19 (or a pharmaceutically acceptable salt thereof) for treating generalized anxiety disorder (GAD) in the absence of mesembrine or mesembrenone.

[0058] Pharmaceutically acceptable carriers include those known in the art. The choice of a pharmaceutically acceptable carrier can depend, for example, on the desired route of administration of the composition. Pharmaceutical compositions (preparations) can be administered to a subject by any of a number of routes, including, for example, parenteral administration (e.g., intravenous, subcutaneous, or intramuscular), oral administration (e.g., tablets and capsules); absorption through oral mucosa (e.g., sublingual) or transdermal (e.g., as a patch applied to the skin), or topical (e.g., as a cream, ointment, or spray applied to the skin).

[0059] In some embodiments, pharmaceutical compositions containing Compound 18 and / or Compound 19, or pharmaceutically acceptable salts thereof, can be formulated for oral administration. For example, the compounds provided herein can be combined with suitable defined excipients to form an oral unit dosage form, such as a capsule or tablet, containing a target dose of mesembranol. The pharmaceutical product can be prepared by first manufacturing mesembranol as the active ingredient (API), followed by roller compaction / milling with intragranular excipients and blending with extragranular excipients. The pharmaceutical product can contain mesembranol as the API and excipient components in a tablet at the desired dosage strength. The blended material can be compressed to form a tablet and then film-coated. Excipients can be selected from materials appropriate for inclusion in a pharmaceutical composition for the intended purpose and delivery route, including those that provide the pharmaceutical composition with desired manufacturing and stability characteristics, and / or desired in vivo or other properties. In some embodiments, the pharmaceutical composition may include mesembranol as the API in combination with a filler (e.g., a form of microcrystalline cellulose), a dry binder, or disintegrant (e.g., a cross-linked polymer), a flow agent (e.g., colloidal silicon dioxide), and / or a lubricant (e.g., magnesium stearate). In some embodiments, the pharmaceutical composition may include materials, such as sustained-release or disintegrants, that are involved in carrying or transporting the API pharmaceutical from one organ or body part of a subject to another, including materials that desirably control absorption of the API in the intestine.

[0060] The formulations can be conveniently presented in unit dosage form and prepared by any method well known in the art of pharmacy. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the host being treated and the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound that produces a therapeutic effect. For use in the methods of the present invention, the active compound can be provided per se, or can be provided as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably 0.5 to 90%) of the active ingredient in combination with a pharmaceutically acceptable carrier.

[0061] Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound of the present invention, with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0062] To prepare solid dosage forms for oral administration, the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, (2) binders, (3) wetting agents, (4) disintegrants, (5) dissolution retarders, (6) absorption enhancers, (7) wetting agents, (8) absorbing agents, (9) lubricants, (10) complexing agents, and (11) colorants. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets, and pills, the pharmaceutical composition may also contain a buffering agent. Similar types of solid compositions may also be used as fillers in soft- and hard-fill gelatin capsules using appropriate excipients. Pharmaceutical compositions according to the present invention may contain conventional pharmaceutical carriers and / or adjuvants. In some embodiments, pharmaceutical compositions according to the present invention may contain conventional carrier agents, including binders, lubricants, and / or flow agents, selected from products and materials commonly used in the pharmaceutical industry to prepare pharmaceutical compositions for the intended route of administration.

[0063] Tablets may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surface active agents, or dispersing agents. Molded tablets may be made by molding a mixture of the powdered compound moistened with an inert liquid diluent in a suitable machine.

[0064] Liquid dosage forms useful for oral administration comprise a pharmaceutically acceptable carrier and an active ingredient, and are provided as a solid form for reconstitution before administration, or as a liquid (e.g., solution, suspension, or emulsion).In addition to the active ingredient, liquid dosage forms may contain an inert diluent commonly used in the art.For example, the formulation of a pharmaceutically acceptable composition for injection may contain water or an aqueous solution such as physiologically buffered saline, or other solvents or vehicles suitable for the intended route of administration.In some embodiments, the pharmaceutical composition is formulated for parenteral administration.

[0065] The therapeutically effective amount of the pharmaceutical composition can be determined by human clinical trials to determine a safe and effective dose for patients with relevant diagnoses. It is generally understood that the effective amount of the compound may vary depending on the subject's weight, sex, age, and medical history. Other factors that affect the effective amount include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, other types of therapeutic agents administered together with the compound of the present invention. A larger total dose can be delivered by multiple administrations of the pharmaceutical composition at a dose and dose interval determined to be safe and effective for the patient.

[0066] In certain embodiments, the pharmaceutical composition may contain less than about 10%, less than about 5%, less than about 3%, less than about 2%, or less than about 1% by weight of the following alkaloid combinations: (-) mesembrine and mesembrenone, or mesembrenone and mesembrenol, or (-) mesembrine, mesembrenone, and mesembrenol. In certain embodiments, the composition is substantially free of (-) mesembrine, mesembrenone, and mesembrenol.

[0067] In certain embodiments, the pharmaceutical composition comprises a therapeutically effective amount of (-) mesembranol or (-) 6-epi-mesembrananol.

[0068] The present disclosure includes the use of pharmaceutically acceptable salts of the compounds of the present invention in the compositions and methods of the present invention. Pharmaceutically acceptable salts include, for example, acid addition salts and base addition salts. The acid added to a compound to form an acid addition salt can be an organic acid or an inorganic acid. The base added to a compound to form a base addition salt can be an organic base or an inorganic base. In some embodiments, the pharmaceutically acceptable salt is a metal salt, and in some embodiments, the pharmaceutically acceptable salt is an ammonium salt. For example, pharmaceutically acceptable acid addition salts may exist as various solvates with water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates may also be prepared. The source of such solvates may be derived from the solvent of crystallization, may be inherent in the solvent of preparation or crystallization, or may be associated with such solvent.

[0069] In some embodiments, the pharmaceutical composition comprises Compound 018 or a pharmaceutically acceptable salt thereof, and includes excipients containing starch, gelatin, silicone, titanium dioxide, iron oxide, and other inactive ingredients such as color dyes. In some embodiments, a tablet or capsule contains Compound 018, or a pharmaceutical composition thereof, and inactive ingredients.

[0070] In some embodiments, the pharmaceutical composition comprises Compound 019 or a pharmaceutically acceptable salt thereof, and includes excipients containing starch, gelatin, silicone, titanium dioxide, iron oxide, and other inactive ingredients such as color dyes. In some embodiments, a tablet or capsule contains Compound 019, or a pharmaceutical composition thereof, and inactive ingredients.

[0071] Selective inhibition of 5-HTT In some embodiments, mesembranol (compound 18) or a pharmaceutically acceptable salt thereof may be used to inhibit the serotonin transporter (5-HTT) in the absence of mesembrenone and mesembrine without inhibiting phosphodiesterase-4 (PDE4). In some embodiments, (-)6-epi-mesembranol (compound 19) or a pharmaceutically acceptable salt thereof may be used to inhibit the serotonin transporter (5-HTT) in the absence of mesembrine and mesembrenone without inhibiting phosphodiesterase-4 (PDE4).

[0072] For certain compositions described herein, the ratio of PDE4 / SERT inhibition values ​​is calculated by (i) the SERT inhibition value (e.g., IC) of the composition as determined via the assay described in Example 3A. 50 ), and (ii) PDE4 inhibition values ​​(e.g., IC ) for the same compositions determined via the assay described in Example 3B. 50 In certain embodiments, the ratio of PDE4 / SERT inhibition values ​​is at least 10:1, at least 100:1, or at least 500:1. In some embodiments, the pharmaceutical composition has an IC 50 is less than about 30 nM or less than about 15 nM.

[0073] In some embodiments, methods are provided for inhibiting serotonin transporter (5-HTT) in the central nervous system (CNS) of a subject using an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar, the methods comprising administering an API consisting of mesembranol (Compound 18) or a pharmaceutically acceptable salt thereof, (-)6-epi-mesembranol (Compound 19) or a pharmaceutically acceptable salt thereof, or a mixture of Compound 18 and Compound 19 or a pharmaceutically acceptable salt thereof. In some embodiments, the methods for inhibiting serotonin transporter (5-HTT) in the central nervous system (CNS) of a subject using an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar comprise administering an API consisting of mesembranol (Compound 18) or a pharmaceutically acceptable salt thereof. In some embodiments, a method for inhibiting serotonin transporter (5-HTT) in a subject's central nervous system (CNS) using an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar comprises administering an API consisting of (-)6-epi-mesembranol (Compound 19) or a pharmaceutically acceptable salt thereof. In certain embodiments, the API has an inhibition rate of less than about 6% against at least one of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2, at least two of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2, at least three of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2, or against each of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0074] In some embodiments, methods of treatment include administering mesembranol (Compound 18), (-)6-epi-mesembranol (Compound 19), or a pharmaceutically acceptable salt thereof, in the absence of mesembrine and mesembrenone, to inhibit serotonin transporter (5-HTT) without inhibiting phosphodiesterase-4 (PDE4) in a subject in need thereof. In some embodiments, methods of treatment include administering mesembranol (Compound 18), or a pharmaceutically acceptable salt thereof, in the absence of mesembrine and mesembrenone, to inhibit serotonin transporter (5-HTT) without inhibiting phosphodiesterase-4 (PDE4) in a subject in need thereof. In some embodiments, the method of treatment comprises administering (-)6-epi-mesembranol (compound 19) or a pharmaceutically acceptable salt thereof, in the absence of mesembrine and mesembrenone, to inhibit the serotonin transporter (5-HTT) in a subject in need of such inhibition without inhibiting phosphodiesterase-4 (PDE4). In certain embodiments, mesembranol (compound 18), (-)6-epi-mesembranol (compound 19), or a pharmaceutically acceptable salt thereof, has an inhibition rate of less than about 6% against at least one of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2, and against at least two of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2, against at least three of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2, or against each of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0075] In some embodiments, a method for inhibiting serotonin transporter (5-HTT) in a subject in need thereof using an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar comprises administering to the subject an API comprising mesembranol (Compound 18), (-)6-epi-mesembranol (Compound 19), or a pharmaceutically acceptable salt thereof. In some embodiments, a method for inhibiting serotonin transporter (5-HTT) in a subject in need thereof using an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar comprises administering to the subject an API comprising mesembranol (Compound 18) or a pharmaceutically acceptable salt thereof. In some embodiments, a method for inhibiting serotonin transporter (5-HTT) in a subject in need thereof with an API that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar comprises administering to the subject an API comprising mesembranol (Compound 18) or a pharmaceutically acceptable salt thereof. In some embodiments, a method for inhibiting serotonin transporter (5-HTT) in a subject in need thereof with an API that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar comprises administering to the subject an API comprising (-)6-epi-mesembranol (Compound 19) or a pharmaceutically acceptable salt thereof.In some embodiments, a method for inhibiting the serotonin transporter (5-HTT) in a subject in need thereof using an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar, the method comprising administering to the subject an API comprising (-)6-epi-mesembranol (Compound 19) or a pharmaceutically acceptable salt thereof. In certain embodiments, the API has an inhibition rate of less than about 6% against at least one of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2, against at least two of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2, against at least three of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2, or against each of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0076] In some embodiments, a method for inhibiting serotonin transporter (5-HTT) in a subject in need thereof with an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar comprises administering to the subject mesembranol (compound 18), (-)6-epi-mesembranol (compound 19), or a pharmaceutically acceptable salt thereof, in the absence of mesembrine or mesembrenone. In some embodiments, a method for inhibiting serotonin transporter (5-HTT) in a subject in need thereof using an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar comprises administering mesembranol (Compound 18), or a pharmaceutically acceptable salt thereof, to the subject in the absence of mesembrine or mesembrenone. In some embodiments, a method for inhibiting serotonin transporter (5-HTT) in a subject in need thereof using an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar comprises administering mesembranol (Compound 18), or a pharmaceutically acceptable salt thereof, to the subject in the absence of mesembrine. In some embodiments, a method for inhibiting serotonin transporter (5-HTT) in a subject in need of inhibition of the serotonin transporter (5-HTT) using an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar, the method comprising administering to the subject mesembranol (compound 18), or a pharmaceutically acceptable salt thereof, in the absence of mesembrenone.In some embodiments, a method for inhibiting serotonin transporter (5-HTT) in a subject in need thereof using an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar, the method comprising administering to the subject (-)6-epi-mesembranol (compound 19), or a pharmaceutically acceptable salt thereof, in the absence of mesembrine and mesembrenone. In some embodiments, a method for inhibiting serotonin transporter (5-HTT) in a subject in need thereof using an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar, the method comprising administering to the subject (-)6-epi-mesembranol (compound 19), or a pharmaceutically acceptable salt thereof, in the absence of mesembrine. In some embodiments, a method for inhibiting serotonin transporter (5-HTT) in a subject in need of inhibition of serotonin transporter (5-HTT) using an active ingredient (API) that inhibits PDE4A1A, PDE4B2, PDE4C1, and PDE4D2 by about 5% or less at 10 micromolar comprises administering to the subject (-)6-epi-mesembranol (compound 19), or a pharmaceutically acceptable salt thereof, in the absence of mesembrenone. In certain embodiments, the API has an inhibition rate of less than about 6% against at least one of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2, against at least two of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2, against at least three of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2, or against each of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0077] In some embodiments, methods of treatment are provided that include administering a pharmaceutical composition comprising an API consisting of compound 18 (or a pharmaceutically acceptable salt thereof) and / or compound 19 (or a pharmaceutically acceptable salt thereof).

[0078] In some embodiments, methods of treating generalized anxiety disorder are provided. Generalized anxiety disorder (DSM-IV) is characterized by excessive anxiety or worry (anticipatory anxiety) that persists for at least six months and that the individual finds difficult to control. It must be associated with at least three of the following symptoms: restlessness, tension or apprehension, feeling tired easily, difficulty concentrating or disorganized thinking, irritability, muscle tension, and sleep disturbances. In some embodiments, a method of treating generalized anxiety disorder (GAD) comprises administering a pharmaceutical composition comprising compound 18 (or a pharmaceutically acceptable salt thereof) as an API. In some embodiments, a method of treating generalized anxiety disorder (GAD) comprises administering a pharmaceutical composition comprising compound 19 (or a pharmaceutically acceptable salt thereof) as an API. In some embodiments, a method of treating generalized anxiety disorder (GAD) comprises administering a pharmaceutical composition comprising a mixture of compound 18 (or a pharmaceutically acceptable salt thereof) and compound 19 (or a pharmaceutically acceptable salt thereof) as APIs. In some embodiments, a method for treating generalized anxiety disorder (GAD) comprises administering a pharmaceutical composition comprising compound 18 (or a pharmaceutically acceptable salt thereof) as an API in the absence of mesembrine or mesembrenone. In some embodiments, a method for treating generalized anxiety disorder (GAD) comprises administering a pharmaceutical composition comprising compound 18 (or a pharmaceutically acceptable salt thereof) as an API, wherein the pharmaceutical composition comprises 0-0.5% mesembrine or mesembrenone as measured by HPLC. In some embodiments, a method for treating generalized anxiety disorder (GAD) comprises administering a pharmaceutical composition comprising compound 19 (or a pharmaceutically acceptable salt thereof) as an API, wherein the pharmaceutical composition comprises 0-0.5% mesembrine or mesembrenone as measured by HPLC. In some embodiments, a method for treating generalized anxiety disorder (GAD) comprises administering a pharmaceutical composition comprising a mixture of compound 18 (or a pharmaceutically acceptable salt thereof) and compound 19 (or a pharmaceutically acceptable salt thereof) as active ingredients (APIs), wherein the pharmaceutical composition comprises 0-0.5% mesembrine or mesembrenone as measured by HPLC.

[0079] definition Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings commonly understood by those of ordinary skill in the art. Generally, the nomenclature used in connection with, and techniques of, chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry described herein are those well known and commonly used in the art.

[0080] The methods and techniques of the present disclosure, unless otherwise indicated, are generally performed according to conventional methods well known in the art and as described in various general and more specific references cited and discussed throughout this specification. See, for example, "Principles of Neural Science," McGraw-Hill Medical, New York, NY (2000); Motulsky, "Intuitive Biostatistics," Oxford University Press, Inc. (1995); Lodish et al., "Molecular Cell Biology, 4th ed.", W.H. Freeman & Co., New York (2000); Griffiths et al., "Introduction to Genetic Analysis, 7th ed.", W.H. Freeman & Co., NY (1999); and Gilbert et al., "Developmental Biology, 6th ed.", Sinauer Associates, Inc., Sunderland, MA (2000).

[0081] All of the above, as well as any other publications, patents, and published patent applications mentioned in this application, are specifically incorporated herein by reference. In case of conflict, the present specification, including specific definitions, will control.

[0082] The term "agent" is used herein to refer to a chemical compound (e.g., an organic or inorganic compound, a mixture of compounds), a biological macromolecule (e.g., nucleic acids, antibodies containing portions thereof, as well as humanized, chimeric, and human antibodies, and monoclonal antibodies, proteins or portions thereof, e.g., peptides, lipids, carbohydrates), or an extract prepared from biological materials such as bacteria, plants, fungi, or animal (especially mammalian) cells or tissues. Agents include, for example, agents of known structure and agents of unknown structure.

[0083] The terms "patient," "subject," or "individual" are used interchangeably and refer to either a human or non-human animal. These terms include mammals, such as humans, primates, livestock animals (including cows and pigs), companion animals (e.g., dogs and cats), and rodents (e.g., mice and rats).

[0084] "Treating" a condition or patient refers to taking measures to obtain beneficial or desired results, including clinical results. As used herein, and as well understood in the art, "treatment" is an approach for obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, whether detectable or undetectable, reduction in the extent of disease, stabilization of the disease state (i.e., not worsening), prevention of disease spread, delay or slowing of disease progression, improvement or palliation of the disease state, and remission (whether partial or total). "Treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment.

[0085] The term "prevention," when used in reference to a condition such as local recurrence (e.g., pain), a disease such as cancer, a complex syndrome such as heart failure, or any other medical condition, is art-recognized and well understood in the art and includes administration of a composition that reduces the frequency of or delays the onset of symptoms of a medical condition in a subject compared to subjects not administered the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous growths in a population of patients receiving prophylactic treatment compared to an untreated control population, and / or delaying the appearance of detectable cancerous growths in a treated population relative to an untreated control population by a statistically and / or clinically significant amount.

[0086] "Administering" or "administration" of a substance, compound, or agent to a subject can be accomplished using one of a variety of methods known in the art. For example, the compound or agent can be administered intravenously, intraarterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ophthalmically, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, and transdermally (by absorption, e.g., through the skin tract). The compound or agent can also be suitably introduced by refillable or biodegradable polymeric or other devices, e.g., patches and pumps, or formulations that provide sustained, slow, or controlled release of the compound or agent. Administration can also be, for example, once, multiple times, and / or over one or more extended periods of time.

[0087] Suitable methods of administering a substance, compound, or agent to a subject also depend, for example, on the age and / or physical condition of the subject, as well as the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability, and toxicity). In some embodiments, the compound or agent is administered to the subject orally, e.g., by ingestion. In some embodiments, the orally administered compound or agent is in a sustained- or slow-release formulation or is administered using such a slow- or sustained-release device.

[0088] As used herein, the term "co-administration" refers to any administration form of two or more different therapeutic agents in which a second agent is administered while a previously administered therapeutic agent is still effective in the body (e.g., the two agents are effective in a patient simultaneously, which may include a synergistic effect of the two agents). For example, different therapeutic compounds can be administered simultaneously or sequentially in the same formulation or in separate formulations. Thus, an individual receiving such treatment can benefit from the combined effects of the different therapeutic agents.

[0089] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or agent is an amount of drug or agent that has the intended therapeutic effect when administered to a subject. The full therapeutic effect does not necessarily occur in a single administration, but may occur only after a series of administrations. Thus, a therapeutically effective amount can be administered in one or more administrations. The exact effective amount required by a subject will depend, for example, on the subject's size, health, and age, as well as the nature and extent of the condition being treated, such as cancer or MDS. Those skilled in the art can easily determine the effective amount for a given situation by routine experimentation.

[0090] As used herein, the term "optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes examples of when the event or circumstance occurs and examples of when it does not occur. For example, "optionally substituted alkyl" refers to cases where alkyl can be substituted, as well as cases where alkyl is not substituted.

[0091] It is understood that the substituents and substitution patterns on the compounds of the present invention can be selected by one skilled in the art to result in chemically stable compounds that can be readily synthesized from readily available starting materials by techniques known in the art, as well as those methods described below. When a substituent itself is substituted with two or more groups, it is understood that these multiple groups can be on the same carbon or on different carbons, so long as a stable structure results.

[0092] As used herein, the term "modulate" includes inhibiting or suppressing a function or activity (eg, cell proliferation) and enhancing a function or activity.

[0093] "Pharmaceutically acceptable salt" or "salt" is used herein to refer to an acid addition salt or a base addition salt that is suitable or compatible for the treatment of patients.

[0094] As used herein, the term "pharmaceutically acceptable acid addition salt" refers to any non-toxic organic or inorganic salt of any of the principal ingredients of the compounds described herein. Exemplary inorganic acids that form suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, as well as metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Exemplary organic acids that form suitable salts include mono-, di-, and tricarboxylic acids such as glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, benzoic acid, phenylacetic acid, cinnamic acid, and salicylic acid, as well as sulfonic acids such as p-toluenesulfonic acid and methanesulfonic acid. Mono- or di-acid salts may be formed, and such salts may exist in either hydrated, solvated, or substantially anhydrous form. In general, acid addition salts of the compounds described herein are more soluble in water and various hydrophilic organic solvents and generally exhibit higher melting points than their free base forms. The selection of appropriate salts is known to those skilled in the art. Other pharmaceutically unacceptable salts, such as oxalates, may be used, for example, for laboratory use in isolating the compounds described herein or for subsequent conversion to a pharmaceutically acceptable acid addition salt.

[0095] As used herein, the term "pharmaceutically acceptable base addition salt" refers to any non-toxic organic or inorganic base addition salt of any acid compound described herein or any of their intermediates. Exemplary inorganic bases that form suitable salts include lithium, sodium, potassium, calcium, magnesium, or barium hydroxide. Exemplary organic bases that form suitable salts include aliphatic, alicyclic, or aromatic organic amines such as methylamine, trimethylamine, and picoline or ammonia. The selection of a suitable salt is known to those skilled in the art.

[0096] As used herein, the phrase "pharmaceutically acceptable" is used to refer to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0097] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.

[0098] As used herein, the phrases "parenteral administration" and "administered parenterally" refer to modes of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravenous, intraocular (e.g., intravitreal), intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration include one or more active compounds in combination with one or more pharmaceutically acceptable sterile, isotonic aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, which may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents, or sterile powders that can be reconstituted into a sterile injectable solution or dispersion immediately before use.

[0099] Many of the compounds useful in the methods and compositions of the present disclosure have at least one asymmetric center in their structure. This stereocenter may exist in the R or S configuration, and the R and S designations are used in accordance with the conventions set forth in Pure Appl. Chem. (1976), 45, 11-30. The present disclosure contemplates all stereoisomeric forms, such as enantiomeric and diastereomeric forms, of the compounds, salts, or mixtures thereof, including all possible mixtures of stereoisomers. See, e.g., WO 01 / 062726.

[0100] Furthermore, certain compounds containing alkenyl groups can exist as Z (Zusammen) or E (Entgegen) isomers, and in each case, the present disclosure includes both mixtures and separate individual isomers.

[0101] Some compounds may also exist in tautomeric forms, and such forms, although not explicitly shown in the formulae given herein, are intended to be included within the scope of the present disclosure.

[0102] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, useful in formulating a drug for pharmaceutical or therapeutic use.

[0103] As used herein, the terms "logarithm of solubility," "LogS," or "logS" are used in the art to quantify the aqueous solubility of a compound. A compound's water solubility significantly affects its absorption and distribution characteristics. Poor solubility often results in poor absorption. The LogS value is the unitless logarithm (base 10) of solubility measured in mol / liter.

[0104] Additional Embodiments 1. A method for treating a psychiatric disorder, comprising administering to a mammal in need thereof an effective amount of a compound selected from (-) mesembranol and (-)6-epi-mesembrananol, or a pharmaceutically acceptable salt thereof.

[0105] 2. The method of embodiment 1, wherein said compound is (-) mesembranol.

[0106] 3. The method of embodiment 1, wherein the compound is (-)6-epi-mesembrananol.

[0107] 4. The method of any one of embodiments 1 to 3, wherein the psychiatric disorder is anxiety, stress, or depression.

[0108] 5. The method of embodiment 4, wherein the psychiatric disorder is an anxiety disorder.

[0109] 6. The method of embodiment 4, wherein the psychiatric disorder is stress.

[0110] 7. The method of embodiment 4, wherein the psychiatric disorder is depression.

[0111] 8. The method of any one of embodiments 1 to 7, wherein the mammal is a human.

[0112] 9. A method for inhibiting SERT, comprising administering to a mammal in need thereof a therapeutically effective amount of a compound selected from (-) mesembranol and (-)6-epi-mesembrananol, or a pharmaceutically acceptable salt thereof.

[0113] 10. The method of embodiment 9, wherein the compound is (-) mesembranol.

[0114] 11. The method of embodiment 9, wherein the compound is (-)6-epi-mesembrananol.

[0115] 12. A method for inhibiting SERT, comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound selected from (-) mesembranol and (-)6-epi-mesembrananol, or a pharmaceutically acceptable salt thereof.

[0116] 13. The method of embodiment 12, wherein the compound is (-) mesembranol.

[0117] 14. The method of embodiment 12, wherein the compound is (-)6-epi-mesembrananol.

[0118] 15. The method of any one of embodiments 12-14, wherein the pharmaceutical composition contains no more than about 10% of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0119] 16. The method of any one of embodiments 12-14, wherein the pharmaceutical composition contains about 5% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0120] 17. The method of any one of embodiments 12-14, wherein the pharmaceutical composition contains about 3% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0121] 18. The method of any one of embodiments 12-14, wherein the pharmaceutical composition contains about 2% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0122] 19. The method of any one of embodiments 12-14, wherein the pharmaceutical composition contains about 1% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0123] 20. The method of any one of embodiments 15-19, wherein the alkaloid is (-) mesembrine.

[0124] 21. The method of any one of embodiments 15-19, wherein the alkaloid is mesembrenone.

[0125] 22. The method of any one of embodiments 15-19, wherein the alkaloid is mesembrenol.

[0126] 23. The method of any one of embodiments 15-19, wherein the pharmaceutical composition is substantially free of (-) mesembrine, mesembrenone, and mesembrenol.

[0127] 24. A pharmaceutical composition comprising (-)6-epi-mesembranol and a pharmaceutically acceptable excipient.

[0128] 25. The pharmaceutical composition of embodiment 24, wherein the pharmaceutical composition contains no more than about 10% of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0129] 26. The pharmaceutical composition of embodiment 24, wherein the pharmaceutical composition contains about 5% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0130] 27. The pharmaceutical composition of embodiment 24, wherein the pharmaceutical composition contains about 3% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0131] 28. The pharmaceutical composition of embodiment 24, wherein less than about 60% of the total alkaloid content is mesembrenone and mesembrenol.

[0132] 29. The pharmaceutical composition of embodiment 24, wherein less than about 20% of the total alkaloid content is (-) mesembrine.

[0133] 30. The pharmaceutical composition of embodiment 24, wherein the composition comprises about 0.3% to about 0.5% by weight of the total alkaloid content, and wherein mesembrenone and mesembrenol are less than about 60% by weight of the total alkaloid content.

[0134] 31. The pharmaceutical composition of embodiment 24, wherein the composition comprises about 0.3% to about 0.6% by weight of the total alkaloid content, and wherein mesembrine is less than about 20% by weight of the total alkaloid content.

[0135] 32. The pharmaceutical composition of embodiment 30 or 31, wherein (-)6-epi-mesembrananol is at least 70% of the total alkaloid content in the composition.

[0136] 33. A pharmaceutical composition according to embodiment 30 or 31, wherein (-)6-epi-mesembrananol is at least 80% of the total alkaloid content in the composition.

[0137] 34. The pharmaceutical composition of embodiment 30 or 31, wherein (-)6-epi-mesembrananol is at least 90% of the total alkaloid content in the composition.

[0138] 35. The pharmaceutical composition of embodiment 24, wherein the composition comprises less than about 70 micrograms of mesembrenone and mesembrenol per 11.3 mg of total alkaloid content.

[0139] 36. The pharmaceutical composition of embodiment 24, wherein the composition comprises less than about 23 micrograms of (-) mesembrine per 11.3 mg of total alkaloid content.

[0140] 37. A pharmaceutical composition according to any one of embodiments 24 to 36, wherein the ratio of PDE4 / SERT inhibition values ​​is at least 10:1.

[0141] 38. A pharmaceutical composition according to any one of embodiments 24 to 36, wherein the ratio of PDE4 / SERT inhibition values ​​is at least 100:1.

[0142] 39. A pharmaceutical composition according to any one of embodiments 24 to 36, wherein the ratio of PDE4 / SERT inhibition values ​​is at least 500:1.

[0143] 40. The pharmaceutical composition of any one of embodiments 24-39, wherein the (-)6-epi-mesembrananol has a half-life of at least 30 minutes in a human hepatocyte assay of the pharmaceutical composition.

[0144] 41. The pharmaceutical composition of any one of embodiments 24-39, wherein the (-)6-epi-mesembrananol has a half-life of at least 60 minutes in a human hepatocyte assay of the pharmaceutical composition.

[0145] 42. The pharmaceutical composition of any one of embodiments 24-39, wherein the (-)6-epi-mesembrananol has a half-life of at least 90 minutes in a human hepatocyte assay of the pharmaceutical composition.

[0146] 43. The pharmaceutical composition of any one of embodiments 24-39, wherein the (-)6-epi-mesembrananol has a half-life of at least 120 minutes in a human hepatocyte assay of the pharmaceutical composition.

[0147] 44. AUC of (-)6-epi-mesembranor vs. (-)mesembrine last 44. The pharmaceutical composition according to any one of embodiments 24 to 43, wherein the ratio is from about 2 to about 2.5.

[0148] 45. A pharmaceutical composition comprising (-) mesembranol and a pharmaceutically acceptable excipient.

[0149] 46. ​​The pharmaceutical composition of embodiment 45, wherein the pharmaceutical composition contains about 10% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0150] 47. The pharmaceutical composition of embodiment 46, wherein the pharmaceutical composition contains about 5% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0151] 48. The pharmaceutical composition of embodiment 46, wherein the pharmaceutical composition contains about 3% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

[0152] 49. The pharmaceutical composition of embodiment 46, wherein the pharmaceutical composition contains less than about 60% mesembrenone and mesembrenol.

[0153] 50. The pharmaceutical composition of embodiment 46, wherein the pharmaceutical composition contains less than about 20% (-) mesembrine.

[0154] 51. The pharmaceutical composition of embodiment 46, wherein the composition comprises about 0.3% to about 0.5% by weight of the total alkaloid content, and wherein mesembrenone and mesembrenol are less than about 60% by weight of the total alkaloid content.

[0155] 52. The pharmaceutical composition of embodiment 46, wherein the composition comprises about 0.3% to about 0.6% by weight of the total alkaloid content, and wherein mesembrine is less than about 20% by weight of the total alkaloid content.

[0156] 53. A pharmaceutical composition according to embodiment 51 or 52, wherein (-) mesembranol is at least 70% of the total amount of alkaloids in the composition.

[0157] 54. A pharmaceutical composition according to embodiment 51 or 52, wherein (-) mesembranol is at least 80% of the total alkaloid content in the composition.

[0158] 55. A pharmaceutical composition according to embodiment 51 or 52, wherein (-) mesembranol is at least 90% of the total alkaloid content in the composition.

[0159] 56. The pharmaceutical composition of embodiment 46, wherein the composition comprises less than about 70 micrograms of mesembrenone and mesembrenol per 11.3 mg of total alkaloid content.

[0160] 57. The pharmaceutical composition of embodiment 46, wherein the composition comprises less than about 23 micrograms of (-) mesembrine per 11.3 mg of total alkaloid content.

[0161] 58. A pharmaceutical composition according to any one of embodiments 45 to 57, wherein the ratio of PDE4 / SERT inhibition values ​​is at least 10:1.

[0162] 59. A pharmaceutical composition according to any one of embodiments 45 to 57, wherein the ratio of PDE4 / SERT inhibition values ​​is at least 100:1.

[0163] 60. A pharmaceutical composition according to any one of embodiments 45 to 57, wherein the ratio of PDE4 / SERT inhibition values ​​is at least 500:1.

[0164] 61. The pharmaceutical composition according to any one of embodiments 45 to 60, wherein the (-)6-epi-mesembrananol has a half-life of at least 30 minutes in a human hepatocyte assay of the pharmaceutical composition.

[0165] 62. The pharmaceutical composition of any one of embodiments 45-60, wherein the (-)6-epi-mesembrananol has a half-life of at least 60 minutes in a human hepatocyte assay of the pharmaceutical composition.

[0166] 63. The pharmaceutical composition according to any one of embodiments 45 to 60, wherein the (-)6-epi-mesembrananol has a half-life of at least 90 minutes in a human hepatocyte assay of the pharmaceutical composition.

[0167] 64. The pharmaceutical composition according to any one of embodiments 45 to 60, wherein the (-)6-epi-mesembrananol has a half-life of at least 120 minutes in a human hepatocyte assay of the pharmaceutical composition.

[0168] 65. AUC of (-)6-epi-mesembranor vs. (-)mesembrine last The pharmaceutical composition according to any one of embodiments 45 to 64, wherein the ratio is from about 2 to about 2.5.

[0169] 66. A method for inhibiting SERT without inhibiting PDE4, comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition according to any one of embodiments 24 to 65.

[0170] 67. The method of embodiment 66, wherein the pharmaceutical composition comprises (-) mesembranol.

[0171] 68. The method of embodiment 66, wherein the pharmaceutical composition is (-)6-epi-mesembrananol.

[0172] 69. The method of any one of embodiments 66-68, wherein the pharmaceutical composition has an IC50 against SERT of less than about 30 nM.

[0173] 70. The pharmaceutical composition comprises an IC inhibitor against SERT. 50 69. The method of embodiment 66 or 68, wherein the

[0174] 71. The method of any one of embodiments 66-70, wherein the pharmaceutical composition has an inhibition rate of less than about 6% against at least one of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0175] 72. The method of any one of embodiments 66-70, wherein the pharmaceutical composition has an inhibition rate of less than about 6% against at least two of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0176] 73. The method of any one of embodiments 66-70, wherein the pharmaceutical composition has an inhibition rate of less than about 6% against at least three of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0177] 74. The method of any one of embodiments 66 to 70, wherein the pharmaceutical composition has an inhibition rate of less than about 6% against each of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0178] 75. The method according to any one of embodiments 66 to 74, wherein the ratio of PDE4 / SERT inhibition values ​​is at least 10:1.

[0179] 76. The method according to any one of embodiments 66 to 74, wherein the ratio of PDE4 / SERT inhibition values ​​is at least 100:1.

[0180] 77. The method according to any one of embodiments 66-74, wherein the ratio of PDE4 / SERT inhibition values ​​is at least 500:1.

[0181] 78. A method for inhibiting SERT without inhibiting PDE4, comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition.

[0182] 79. The method of embodiment 78, wherein the pharmaceutical composition is a composition described in any one of embodiments 24 to 65.

[0183] 80. The pharmaceutical composition comprises an IC inhibitor against SERT. 50 80. The method of embodiment 78 or 79, wherein the IL-10 is less than about 30 nM.

[0184] 81. The pharmaceutical composition comprises an IC inhibitor against SERT. 50 80. The method of embodiment 78 or 79, wherein the IL-10 is less than about 15 nM.

[0185] 82. The method of any one of embodiments 78-81, wherein the pharmaceutical composition has an inhibition rate of less than about 6% against at least one of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0186] 83. The method of any one of embodiments 78-81, wherein the pharmaceutical composition has an inhibition rate of less than about 6% against at least two of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0187] 84. The method of any one of embodiments 78-81, wherein the pharmaceutical composition has an inhibition rate of less than about 6% against at least three of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0188] 85. The method of any one of embodiments 78-81, wherein the pharmaceutical composition has an inhibition rate of less than about 6% against each of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

[0189] 86. The method according to any one of embodiments 78 to 85, wherein the ratio of PDE4 / SERT inhibition values ​​is at least 10:1.

[0190] 87. The method according to any one of embodiments 78 to 85, wherein the ratio of PDE4 / SERT inhibition values ​​is at least 100:1.

[0191] 88. The method according to any one of embodiments 78 to 85, wherein the ratio of PDE4 / SERT inhibition values ​​is at least 500:1.

[0192] 89. A composition comprising mesembranol (compound 18) or a pharmaceutically acceptable salt thereof, wherein the composition contains less than about 0.5% mesembrine or mesembrenone as measured by HPLC.

[0193] 90. The composition of embodiment 89, further comprising (-)6-epi-mesembranol (compound 19) or a pharmaceutically acceptable salt thereof.

[0194] 91. A composition comprising (-)6-epi-mesembranol (compound 19) or a pharmaceutically acceptable salt thereof, said composition containing less than about 0.5% mesembrine or mesembrenone as measured by HPLC.

[0195] 92. The composition of any one of embodiments 89-91, wherein the composition does not contain mesembrine or mesembrenone.

[0196] 93. The composition of any one of embodiments 89-92, wherein the composition does not contain any additional alkaloid compounds other than compound 18 or compound 19.

[0197] 94. The composition of embodiment 89, wherein the composition is a pharmaceutical composition comprising an active ingredient (API) consisting of compound 18 (or a pharmaceutically acceptable salt thereof).

[0198] 95. The composition of embodiment 91, wherein the composition is a pharmaceutical composition comprising an active ingredient (API) consisting of compound 19 (or a pharmaceutically acceptable salt thereof).

[0199] 96. The composition of any one of embodiments 89-93, wherein the composition is a pharmaceutical composition comprising an active ingredient (API) consisting of compound 18 (or a pharmaceutically acceptable salt thereof) and / or compound 19 (or a pharmaceutically acceptable salt thereof).

[0200] 97. A method for treating anxiety, said method comprising administering to a subject in need thereof a pharmaceutical composition comprising mesembranol (or a pharmaceutically acceptable salt thereof) and / or (-)6-epi-mesembrananol (or a pharmaceutically acceptable salt thereof).

[0201] 98. The method of embodiment 97, wherein the active ingredient (API) of the pharmaceutical composition consists of a mixture of mesembranol (or a pharmaceutically acceptable salt thereof) and / or (-)6-epi-mesembranol (or a pharmaceutically acceptable salt thereof).

[0202] 99. The method of embodiment 97, wherein the active ingredient (API) of the pharmaceutical composition consists of mesembranol (or a pharmaceutically acceptable salt thereof).

[0203] 100. The method of embodiment 97, wherein the active ingredient (API) of the pharmaceutical composition consists of a mixture of (-)6-epi-mesembranol (or a pharmaceutically acceptable salt thereof).

[0204] 101. The method of any one of embodiments 97-100, wherein the pharmaceutical composition does not contain mesembrine.

[0205] 102. The method of any one of embodiments 97-101, wherein the pharmaceutical composition does not contain mesembrenone.

[0206] 103. The method of any one of embodiments 97-100, wherein the pharmaceutical composition contains less than 0.5% mesembrine or mesembrenone in the pharmaceutical composition, as detected by HPLC.

[0207] 104. The method of any one of embodiments 97-103, wherein said method is a method for treating generalized anxiety disorder (GAD) in a subject diagnosed with GAD.

[0208] 105. A method for inhibiting the serotonin transporter (5-HTT) in the central nervous system of a subject, comprising the step of administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition comprising mesembranol (or a pharmaceutically acceptable salt thereof) and / or (-)6-epi-mesembrananol (or a pharmaceutically acceptable salt thereof), wherein the pharmaceutical composition does not inhibit PDE4A1A, PDE4B2, PDE4C1 or PDE4D2 by more than 5% at 10 micromolar.

[0209] 106. The method of embodiment 105, wherein the pharmaceutical composition does not contain mesembrine.

[0210] 107. The method of any one of embodiments 105-106, wherein the pharmaceutical composition does not contain mesembrenone.

[0211] 108. The method of embodiment 105, wherein the pharmaceutical composition contains less than 0.5% mesembrine or mesembrenone as detected by HPLC. [Example]

[0212] LC / MS spectra were obtained using an Agilent 1200 / G1956A or a SHIMADZU LCMS-2020. Standard LC / MS conditions were as follows (run time 1.55 min): Acidic conditions: Mobile phase A: 0.0375% TFA in water (v / v). Mobile phase B: 0.01875% TFA in acetonitrile (v / v). Column: Kinetex EVO C18 30*2.1mm, 5μm. Basic conditions: Mobile phase A: 0.025% NH₃·H₂O aqueous solution (v / v). Mobile phase B: acetonitrile. Column: Kinetex EVO C18 2.1 x 30 mm, 5 μm. TIFF2025542199000012.tif179165TIFF2025542199000013.tif97165

[0213] TIFF2025542199000014.tif219165

[0214] Overview of names of mesembranol compounds TIFF2025542199000015.tif39165

[0215] Example 1: Synthesis of (3aS,6R,7aS)-3a-(3,4-dimethoxyphenyl)-1-methyloctahydro-1H-indol-6-ol (018, (-)-mesembranol) [ka] A mixture of 001 (200 mg, 691 μmol) and PtO2 (20.0 mg, 88.0 μmol) in IPA (4 mL) was degassed and purged with N2 three times. The mixture was stirred at 25 °C under an atmosphere of N2 for 16 h. The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by recrystallization from EtOH (1 mL) at 25 °C to give (3aS,6R,7aS)-3a-(3,4-dimethoxyphenyl)-1-methyloctahydro-1H-indol-6-ol (018, (-)-mesembranol) (100 mg, 49%) as a white solid. Melting point: 145.5-146.5 o C. LC-MS (ESI + ) m / z 292.4 (M+H). 1H NMR (400 MHz, CDCl3) δ 6.86 - 6.78 (m, 2H), 6.77 - 6.71 (m, 1H), 3.86 (s, 1H), 3.81 (d, J = 6.8 Hz, 6H), 3.30 (dt, J = 6.8, 9.6 Hz, 1H), 2.83 (s, 1H), 2.40 (s, 3H), 2.33 - 2.20 (m, 1H), 2.09 (dd, J = 2.8, 14.8 Hz, 1H), 1.90 - 1.82 (m, 2H), 1.78 (dd, J = 6.8, 11.6 Hz, 1H), 1.67 - 1.62 (m, 2H), 1.57 (td, J = 2.8, 14.8 Hz, 1H), 1.39 - 1.30 (m, 2H).

[0216] Example 2: Synthesis of (3aS,6S,7aS)-3a-(3,4-dimethoxyphenyl)-1-methyloctahydro-1H-indol-6-ol (019, (-)-6-epi-mesembranol) [ka] To a solution of 001 (2.00 g, 6.91 mmol) and CeCl3·7H2O (3.09 g, 8.29 mmol, 788 μL) in MeOH (80 mL) was added NaBH4 (1.57 g, 41.4 mmol). The mixture was stirred at 0 °C for 2 h. The reaction mixture was added to 50 mL of aqueous NH4Cl solution, the organic and aqueous layers were separated, and the aqueous solution was extracted with DCM (50 mL × 3). The organic solutions were combined, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Welch Xtimate C18 150*25mm*5um; mobile phase: [water (NH3H2O)-ACN]; B%: 28%~58%, 8 min) to give (3aS,6S,7aS)-3a-(3,4-dimethoxyphenyl)-1-methyloctahydro-1H-indol-6-ol (019, (-)-6-epimecembranol) (730 mg, 37%) as a white oil. 1 H NMR(400MHz,CDCl3)δ6.95-6.88(m,2H),6.86-6.80(m,1H),3.95(s,1H),3.90(d,J=6.8Hz,6H),3.46-3.35(m,1H),2.93(s,1H),2.50(s,3H) ),2.45-2.29(m,2H),2.19(dd,J=2.4,14.9Hz,1H),2.01-1.82(m,2H),1.79-1.72(m,1H),1.70-1.59(m,3H),1.44(tt,J=2.8,13.6Hz,1H).

[0217] Assay procedure Example 3A: SERT Inhibition Assay SERT inhibition was measured using a neurotransmitter transport fluorescence assay. Briefly, stable 5HTT HEK293 cells were prepared in 384-microwell plates (20,000 cells per well, 20 μL). Compounds were prepared at a maximum concentration of 1 μM in assay buffer (20 mM HEPES, 0.1% BSA in HBSS). Ten doses of test compound (3-fold serial dilutions) were added to the plated cells and incubated at 37°C for 30 minutes. Dye solution (Molecular Devices Neurotransmitter Transporter Uptake Assay Kit) was added at 25 μL per well and incubated at 37°C for 30 minutes. The plates were then read on a plate reader, and the results are shown in Table 1 (n = 6 ± SD).

[0218] [Table 1]

[0219] Example 3B: PDE4 Inhibition Assay Assay conditions for determining PDE4 inhibition rate: A 100 μM dilution of test compound was prepared in assay buffer (10% DMSO concentration), and 5 μl of the dilution was added to a 50 μl reaction mixture, resulting in a final 1% DMSO concentration in all reactions. Enzyme reactions were performed for 60 minutes at room temperature in a 50 μl mixture containing PDE assay buffer, 100 nM FAM-cAMP, PDE enzyme (Table 2.3.1), and test compound (Section 2.2). After the enzymatic reaction, 100 μl of binding solution (1:100 dilution of binder in binder diluent) was added to each reaction, and the reaction was performed for 60 minutes at room temperature.

[0220] Fluorescence intensity was measured using a Tecan Infinite M1000 microplate reader at an excitation wavelength of 485 nm and an emission wavelength of 528 nm.

[0221] PDE4 assay kit (BPS Bioscience, San Diego, CA) as described below.

[0222] Step 1: 1) Dilute the 20 μM FAM-Cyclic-3′,5′-AMP stock solution 100-fold with PDE buffer to prepare a 200 nM solution. 2) Add 25 μl of FAM-Cyclic-3',5'-AMP (200 nM) to each well designated as "positive control," "test inhibitor," and "substrate control." 3) 20 μl of PDE assay buffer was added to each well designated as "substrate control" and 45 μl of PDE assay buffer was added to each well designated as "blank." 4) 5 μl of inhibitor solution was added to each well designated "Test Inhibitor." For wells labeled "Positive Control," "Substrate Control," and "Blank," 5 μl of the same solution without inhibitor (inhibitor buffer) was added. 5) Thaw the PDE on ice. Upon initial thawing, briefly spin the tube containing the enzyme to recover the entire contents of the tube. 6) PDE4 is diluted to 7.5 pg / μl (0.15 ng / reaction) in PDE buffer. The reaction was initiated by adding 20 μl of PDE4 (7.5 pg / μl) to wells designated "positive control" and "test inhibitor." 7) Incubate at room temperature for 1 hour.

[0223] Step 2: 1) Mix the binder thoroughly and dilute the binder 1:100 with the binder diluent. 2) Add 100 μl of diluted binding agent to each microwell. Incubate for 1 hour at room temperature with gentle shaking. 3) Read the fluorescence polarization of the samples using a microtiter plate reader equipped with a fluorescence polarization measurement function capable of excitation at wavelengths of 485 ± 5 nm and emission at wavelengths of 528 ± 10 nm. Subtract the blank value from all other values. 4) Data analysis: PDE activity assays were performed in duplicate at each concentration. Tecan Magellan 6 software was used to convert fluorescence intensity to fluorescence polarization. Fluorescence polarization data were analyzed using the computer software Graphpad Prism. Fluorescence polarization in the absence of compound (FP) for each data set was calculated. t ) was defined as 100% activity. In the absence of PDE and compound, the fluorescence polarization (FP b ) was defined as 0% activity. The activity rate in the presence of compound was calculated according to the following formula: Activity rate (%) = (FP - FP b ) / (FP t -FP b ) × 100% (where FP = fluorescence polarization in the presence of compound). The activity % and compound concentration series values ​​were calculated using the formula Y = B + (TB) / 1 + 10 ((LogEC50-X)×Hill Slope) The IC was plotted using nonlinear regression analysis for sigmoidal dose-response curves generated by the formula (where Y = % activity, B = minimum activity, T = maximum activity, X = logarithm of the compound, and Hill slope = slope coefficient or Hill coefficient). 50 The values ​​were determined by the concentration that caused half-maximal activity. The results are shown in Tables 2A and 2B.

[0224] [Table 2A] [Table 2B]

[0225] Example 4: Hepatocyte metabolism Figure 2A is a graph comparing the compound stability of (-) mesembrine (triangles) and (-) mesembranol (squares) in human hepatocytes as measured according to Example 4. Figure 2B is a graph comparing the compound stability of (-) mesembrine (triangles) and (-) 6-epi-mesembranol (circles) in human hepatocytes as measured according to Example 4.

[0226] A 2.5 μL aliquot of 100 μM test compound was incubated with 247.5 μL of cryopreserved hepatocytes suspended in serum-free culture medium at 1 million viable cells / mL. The mixture was incubated at 37°C and shaken at 500 rpm at designated time points (0.5, 5, 10, 15, 30, 60, 90, or 120 minutes). At each time point, a 25 μL aliquot of the mixed culture was transferred to 125 μL of cold acetonitrile containing an internal standard, followed by centrifugation at 3,220 g for 30 minutes. 100 μL of the supernatant was mixed with 100 μL of distilled water and analyzed by LC-MS / MS. Peak area ratios were calculated from extracted ion chromatograms, and the recovery rates were calculated. The in vitro half-life (t 1 / 2 ) was calculated from the concentration versus time regression slope and the in vitro t 1 / 2 =0.693 / k. in vitro t 1 / 2 In vitro intrinsic clearance (in vitro CL) int , μL / min / 10 6 For conversion to in vitro CL (cells), use the following formula: int = kV / N. The results are also shown in Figures 2A and 2B.

[0227] [Table 3]

[0228] In human hepatocytes, mesembrine (001) undergoes rapid metabolism (t 1 / 2 = 13 min). In comparison, both mesembranol (018) and 6-epi-mesembranol (019) are highly stable in human hepatocytes (t 1 / 2 These data indicate that oral doses of mesembrine undergo rapid first-pass hepatic metabolism in vivo, resulting in low drug concentrations in plasma and brain. Furthermore, residual mesembrine concentrations in plasma and brain are rapidly eliminated, limiting the duration of action. In contrast, oral delivery of mesembranol (018) and 6-epi-mesembranol (019) may result in low hepatic clearance in humans, allowing for stable drug concentrations and a longer duration of action suitable for therapeutic efficacy.

[0229] In vitro intrinsic clearance data in human hepatocytes are predictors of in vivo plasma clearance (drug stability). If a given compound has a high extraction efficiency (maximum theoretical value = 1.0), the drug will be rapidly excreted from the body by the liver. Compound 001 has a high extraction efficiency (E~0.9), while compounds 018 and 019 have low extraction efficiency (E~0.1). As predictors of in vivo drug clearance from plasma, these data indicate that 001 is rapidly eliminated in human plasma in vivo.

[0230] Example 5: Pharmacokinetic study of mesembranol, 6-epi-mesembranol, and mesembrine after PO administration in male SD rats Three separate pharmacokinetic studies were performed for each compound. Generally, 10 mg / kg of compound was administered PO to male SD rats (formulated in 1% NMP and 99% saline v / v). Plasma samples were collected 0 to 4 hours post-dose (n=3 per time point) to measure plasma drug concentrations. Plasma samples were mixed with acetonitrile containing an internal standard. Samples were vortexed and then centrifuged at 4°C for 15 minutes. The supernatant was diluted with water (1:2 V / V) and analyzed by LC / MS / MS for quantitative analysis. Plasma compound concentrations were calculated for each time point.

[0231] [Table 4]

[0232] The data presented in Table 4 are presented as the ratio of values ​​obtained for mesembranol (018): mesembrine (001) ("018:001 ratio"), or (-)-6-epimesembranol (019): (-) mesembrine (001) ("019:001 ratio"). Ratios greater than 1.0 indicate a higher t value in plasma compared to compound 001 (mesembrine). 1 / 2 , C max , or an increase in the area under the curve (AUC). Both Compound 018 and Compound 019 exhibited a shorter plasma half-life (t 1 / 2 ), maximum concentration (C max ), and significantly improved area under the curve (AUC). These results demonstrate improved pharmacokinetic profiles of Compounds 018 and 019 (compared to Compound 001), resulting in elevated plasma levels over a longer period of time. For example, Compound 018 unexpectedly exhibited an approximately three-fold longer plasma half-life, an approximately nine-fold higher Cmax, and an approximately four-fold higher AUC(last) compared to mesembrine (Compound 001). Furthermore, Compound 019 unexpectedly exhibited an approximately two-fold longer plasma half-life, an approximately two-fold higher Cmax, and an approximately two-fold higher AUC(last) compared to mesembrine (Compound 001).

[0233] Example 6: HPLC method for detecting mesembrenol or mesembrenone Gericke et al. (Journal of Ethnopharmacology, 2022, 284, 114550) performed alkaloid profiling and quantification on an ethanol extract of Zembrin® using a validated ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS-PDA) coupled with a photodiode array detector, according to a previously described method (Zhao et al., Phytochemistry, 2018, 152, 191-203). The sample contained mesembrenone (47.9%), mesembrenol (32%), mesembrine (13.3%), and mesembranol (6.8%), in descending order, contributing 3.84 μg / mg of total plant material. Using 25 mg of zebrin (0.4% w / w total alkaloids) as a model extract, this corresponds to an administered dose of approximately 7 micrograms of mesembranol. It is less than 200 nanograms / kg in humans, which is the IC 50 is several orders of magnitude lower than 。

[0234] Example 7: Pharmacokinetic profiles of compounds 018 and 019 in dogs Pharmacokinetic profiles of Compound 018 and Compound 019 evaluated in dogs. Figure 3 compares the plasma concentrations (nM) of Compound 018, Compound 019, and Compound 001 after IV administration of 2 mg / kg of the compounds. Figure 4 compares the plasma concentrations (nM) of Compound 018, Compound 019, and Compound 001 after PO administration of 10 mg / kg of the compounds.

[0235] HPLC equipment: Shimadzu (DGU-20A5R, serial number: L20705621435 IX; LC-30AD serial numbers: L20555611905 AE and L20555611816 AE; SIL-30AC, serial number: L20565605499 AE; Rack Changer II serial number: L20585601125 SS; CTO-20A: serial number: L20205620413 CD; CBM-20A: serial number: L20235636200 CD) MS: AB API 5500 LC / MS / MS instrument (serial number EF221221812) Column: Phenomenex Synergi 2.5 μm Polar-RP 100A (50 × 3 mm) Mobile phase: (A) 5% acetonitrile in water (0.1% formic acid), (B) 95% acetonitrile in water (0.1% formic acid) Sample: 50 μL plasma sample + 5 μL blank solution + 200 μL acetonitrile for PPE (protein precipitation extraction).

[0236] TIFF2025542199000023.tif78165TIFF2025542199000024.tif72165

[0237] Sample preparation The desired serial concentrations of the working solutions were obtained by diluting the analyte stock solution with 50% aqueous acetonitrile. Five µL of working solution (5, 10, 20, 50, 100, 500, 1000, 5000, and 10,000 ng / mL) was added to 50 µL of blank Beagle dog plasma to achieve calibration standards from 0.5 to 1000 ng / mL (0.5, 1, 2, 5, 10, 50, 100, 500, and 1000 ng / mL) in a total volume of 55 µL. Five quality control samples were prepared for plasma at concentrations of 1 ng / mL, 2 ng / mL, 5 ng / mL, 50 ng / mL, and 800 ng / mL, independent of those used for the calibration curve. These QC samples were prepared on the day of analysis using the same method as the calibration standards.

[0238] 55 μL of the standard, 55 μL of the QC sample, and 55 μL of the unknown sample (50 μL of plasma and 5 μL of blank solution) were each added to 200 μL of the acetonitrile-containing IS mixture to precipitate proteins. The samples were then vortexed for 30 seconds. After centrifugation at 3900 rpm for 15 minutes at 4°C, the supernatants were diluted 3-fold with water. 2 μL of the diluted supernatants for SNTX-004 and SNTX-005, and 4 μL for SNTX-001, were injected into the LC / MS / MS system for quantitative analysis.

[0239] Compound information Compound ID Compound 018 MW (free form) 291.39 FW (salt form) 291.39

[0240] Preparation of formulations Preparation of Compound 018 PO (2 mg / kg, 5 mL / kg) Dosing 0.4mg / mL solution of 1% NMP and 99% v / v saline 71.21 mg of SNTX-004 was dissolved in 1.780 mL of NMP using vortexing and sonication, and then 176.245 mL of saline was added using vortexing and sonication to obtain a solution.

[0241] Dog PK data (IV) from compounds 018, 019, and 001 (data plotted in Figure 3) TIFF2025542199000025.tif86165

[0242] Dog PK data (PO) from compounds 018, 019, and 001 (data plotted in Figure 4) TIFF2025542199000026.tif109165

[0243] Example 8: Antidepressant signatures of Compound 018 and Compound 019 in mice using the Smartcube system. To confirm the antidepressant effects of Compounds 018 and 019, the SmartCube® system (Psychogenics, Inc., Paramus, NJ) was used. This system classifies compounds with therapeutic effects for neuropsychiatric conditions using features derived from mouse behavioral data. These features are classified by comparing them with a reference database of unique behavioral feature sets associated with known classes of commercially available drugs used to treat neuropsychiatric conditions. Therefore, this system can be used as a model to identify the psychiatric effects of compounds by comparing their effects with drugs with known, validated effects. By comparing the animal's response to known drugs, test drugs can be classified according to their function, e.g., hallucinogens, anxiolytics, analgesics, cognition enhancers, psychostimulants, mood stabilizers, high-dose antipsychotics, antipsychotics, sedatives / hypnotics, anxiolytics, high-dose antidepressants, and antidepressants.

[0244] Once all features are extracted from the raw data through an automated pipeline, a proprietary bioinformatics algorithm is used to de-correlate groups of features and find the combination of values ​​that best separates different groups of subjects. For each compound, at each dose, the system provides the probability that the drug is active and classifies such estimated activity into different classes of interest.

[0245] The SmartCube® system is designed to successfully measure numerous spontaneous behaviors and responses to tasks in the same testing environment.

[0246] The hardware includes a force sensor and several aversive stimuli to elicit behavior. Three high-resolution video cameras continuously record 3D images of the mouse inside the SmartCube® device (SC) throughout the entire test period.

[0247] Mice were tested in the SmartCube® system and exposed to a series of challenges during a 45-minute test period, with the cube cleaned between each run.

[0248] For class and subclass analysis, a reference dataset was constructed from several hundred drug doses across multiple drug classes and control groups. Each reference drug was tested in mice at multiple doses appropriate for that drug. The best-performing classifier was selected from our evaluation tests, and two different types of classifiers were constructed that make independent predictions at the drug class and subclass levels. Classes consist of drugs currently on the market or drugs clinically validated for specific indications. Subclasses are a larger set than classes, consisting of both marketed drugs and other mechanistically validated compounds.

[0249] The data were processed using proprietary computer vision and data mining algorithms, and the results were compared to the signatures of reference compounds in our database. The data were subjected to multiple analyses to quantitatively and independently predict drug class and drug subclass. The behavioral signatures of test drugs were evaluated using these classifiers to predict potential therapeutic utility.

[0250] A variety of analytical tools are available for follow-up analysis to answer a variety of questions. For example, distinguishing between typical and atypical antipsychotics or similarity to mood stabilizers may be of interest as they broaden the therapeutic indications of lead compounds. While traditional class analysis used in compound screening evaluates 12 pharmacological classes at a time (class analysis), crowdsourcing analysis can narrow the focus to only 2–4 classes of interest. For example, a classifier can be trained using supervised training to separate typical and atypical antipsychotics, and then a novel drug can be used as a test set to classify corresponding samples within the space created by the training. In this way, similarity to one class or the other can be not only quantified but also visualized. As described above, the result of a SmartCube® run is a set of over 2,000 features. By creating independent combinations of the original features, it is possible to reduce the complexity of the data and extract derived features that maximally separate the groups of interest. Similar to principal component analysis, it is possible to find the most significant composite axis that best separates the groups of interest. To create a useful visualization tool, a Gaussian distribution representing the groups of interest can be plotted in the 2D space created by these major axes, where the width is the standard deviation given by the variability between mice. Separation between groups or classes is a measure of similarity or lack thereof (discrimination), and its statistical significance can be calculated by estimating the probability that the results are due to chance.

[0251] Compound 018 was behaviorally inactive (<40%) at 3 mg / kg. At higher doses (10, 30, and 100 mg / kg), the compound exhibited potent antidepressant-like signatures across both class and subclasses, as well as some psychostimulant-like signatures. Subclass analysis showed that the antidepressant signatures were similar to those of SSRIs.

[0252] Compound 019 showed low behavioral activity at 3 mg / kg. At higher doses (10, 30, and 100 mg / kg), the compound exhibited a mixed antidepressant-like and psychostimulant-like signature across both classes and subclasses. The antidepressant signature was similar to that of SSRIs at all doses, and a mixed signature of SSRIs and SNRIs at the 100 mg / kg dose.

Claims

1. 1. A method for treating anxiety or depression comprising administering to a subject in need thereof Compound 019, 【Chemistry 1】 or a pharmaceutically acceptable salt thereof.

2. 10. The method of claim 1, wherein the method is a method for treating major depressive disorder (MDD).

3. 10. The method of claim 1, wherein the method is a method for treating generalized anxiety disorder (GAD).

4. The method of any one of claims 1 to 3, wherein Compound 019, or the pharmaceutically acceptable salt thereof, is administered as a capsule or tablet.

5. 5. The method of claim 4, wherein Compound 019, or the pharmaceutically acceptable salt thereof, is administered once or twice daily.

6. 5. The method of claim 4, wherein the pharmaceutical composition contains about 1% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof.

7. A method for inhibiting SERT comprising administering to a mammal in need thereof a therapeutically effective amount of 【Chemistry 2】 or a pharmaceutically acceptable salt thereof.

8. 1. A pharmaceutical composition comprising: 【Transformation 3】 or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition contains about 3% or less of an alkaloid selected from (-) mesembrine, mesembrenone, mesembrenol, or a combination thereof; The pharmaceutical composition, wherein Compound 019 is at least 90% of the total alkaloid content in the pharmaceutical composition.

9. 9. The pharmaceutical composition of claim 8, wherein the ratio of PDE4 / SERT inhibition value of the pharmaceutical composition is at least 10:

1.

10. 9. The pharmaceutical composition of claim 8, wherein said compound 019 has a half-life of at least 30 minutes in a human hepatocyte assay of said pharmaceutical composition.

11. The pharmaceutical composition is an IC against SERT ５０ The pharmaceutical composition of claim 8, wherein the .alpha.-aspartate is less than about 30 nM.

12. The pharmaceutical composition according to claim 8 or 11, wherein the pharmaceutical composition has an inhibition rate of less than about 6% against at least one of PDE4A1A, PDE4B2, PDE4C1, and PDE4D2.

13. 13. The method of claim 12, wherein the ratio of PDE4 / SERT inhibition value of the pharmaceutical composition is at least 10:

1.

14. 10. A method for inhibiting SERT without inhibiting PDE4, comprising administering a therapeutically effective amount of the pharmaceutical composition of claim 8 to a mammal in need thereof.

15. A composition comprising (-)6-epi-mesembranol or a pharmaceutically acceptable salt thereof, said composition containing less than about 0.5% mesembrine or mesembrenone as measured by HPLC.

16. 16. The composition of claim 15, wherein the composition contains less than about 0.5% (-) mesembranol as measured by HPLC.

17. A composition comprising (-)6-epi-mesembranol, or a pharmaceutically acceptable salt thereof, wherein the composition contains about 0.5% or less of (-)mesembranol as measured by HPLC.

18. A pharmaceutical composition comprising (-)6-epi-mesembrananol or a pharmaceutically acceptable salt thereof, said composition containing less than about 0.5% mesembrine, mesembrenone, or (-)mesembrananol as measured by HPLC.

19. 20. The composition of claim 18 having no more than 0.5% of any other alkaloid compounds as measured by HPLC.

20. 20. A method for treating anxiety or depression, comprising orally administering to a subject in need thereof a therapeutically effective amount of the pharmaceutical composition of any one of claims 17 to 19.

Images