阅读说明：本技术 短效选择性糖皮质激素受体调节剂 (Short acting selective glucocorticoid receptor modulators ) 是由 鲁斯·蒂尔洛夫-埃克尔特 于 2019-12-23 设计创作，主要内容包括：本发明提供了用于预防、缓解或治疗与急性应激或暂时性皮质醇增多症相关的症状、障碍和疾病的短效选择性糖皮质激素受体调节剂,以及包含所述调节剂的立即释放和多次延迟脉冲释放的组合物。(The present invention provides short acting selective glucocorticoid receptor modulators for use in the prevention, alleviation or treatment of symptoms, disorders and diseases associated with acute stress or transient hypercortisolism, as well as compositions comprising immediate release and multiple delayed pulse release of such modulators.)

1. A method of preventing, ameliorating, or treating symptoms, disorders, and diseases associated with acute stress or transient hypercortisolism comprising administering to a subject in need thereof an amount of SASGRM effective to prevent, ameliorate, or treat the symptoms, disorders, and diseases associated with acute stress or transient hypercortisolism in the subject.

2. The method of claim 1, wherein the SASGRM is selected from the group consisting of hydroxy-androst-4, 9(11) -dien-3-one, 21-hydroxy-6, 19-progesterone oxide, 16-hydroxy-11- (substituted phenyl) -estra-4, 9-diene, 17- β -carboxamide for dexamethasone, and Δ 1-11-oxa-11-deoxycorticosterol.

3. The method of claim 1, wherein the SASGRM is selected from the following compounds:

formula I

Formula II

Formula III

Formula IV

And

formula V

Or a combination thereof and pharmaceutically acceptable salts and solvates thereof.

4. The method of claim 1, wherein the symptoms, disorders and diseases associated with acute stress or transient hypercortisolism are selected from academic problems, educational problems, adaptation disorders, stress-related acute insomnia, hypercortisolism-related chronic insomnia accompanied by objective short sleep duration, circadian rhythm disorders, jet-lag circadian rhythm disorders or shift-type circadian rhythm disorders.

5. The method of claim 1, wherein the SASGRM comprises a compound of formula I

Or a pharmaceutically acceptable salt or solvate thereof.

6. The method of claim 1, wherein the SASGRM comprises a compound of formula II

Or a pharmaceutically acceptable salt or solvate thereof.

7. The method of claim 1, wherein the SASGRM comprises a compound of formula III

Or a pharmaceutically acceptable salt or solvate thereof.

8. The method of claim 1, wherein the SASGRM comprises a compound of formula IV

Or a pharmaceutically acceptable salt or solvate thereof.

9. The method of claim 1, wherein the SASGRM comprises a compound of formula V

Or a pharmaceutically acceptable salt or solvate thereof.

10. The method of any one of claims 1-9, wherein the SASGRM is comprised in a composition comprising a plurality of particles.

11. The method of claim 10, wherein the plurality of particles comprise immediate release particles, delayed release particles, pulsed release particles, or a combination thereof.

12. The method of claim 11, wherein the plurality of particles comprises pulsed release particles.

13. The method of claim 12, wherein the pulse-release particles comprise a first population of pulse-release particles that release the SASGRM after a lag period following release of the SASGRM from the immediate-release particles, and a second population of pulse-release particles that release the SASGRM after a lag period following release of the SASGRM from the first population of pulse-release particles.

14. The method of claim 13, wherein the pulse-released particles further comprise a third population of pulse-released particles that release the SASGRM after a lag period following release of the SASGRM from the second population of pulse-released particles.

15. The method of any one of claims 11-14, wherein the delayed release particles and the pulsed release particles comprise a coating.

16. The method of claim 15, wherein the coating comprises a water soluble polymer.

17. The method of claim 16, wherein the water soluble polymer is selected from the group consisting of methylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, cellulose acetate phthalate, polyvinylpyrrolidone/vinyl acetate copolymer, polyvinyl alcohol, polyethylene glycol, polyethylene oxide, hyaluronic acid, alginate, carrageenan, gelatin, and any combination thereof.

18. The method of claim 15, wherein the coating comprises a water insoluble polymer.

19. The method of claim 18, wherein the water insoluble polymer is selected from the group consisting of polyvinyl acetate, cellulose acetate, methyl cellulose, ethyl cellulose, cellulose acetate butyrate, cellulose acetate propionate, non-crystalline cellulose, polyethylene, polyvinyl alcohol, polyacrylates, methacrylates, polyvinyl acetate butyrate, polyvinyl acetate propionate, polyvinyl acetate butyrate, polyvinyl acetate, polyvinyl alcohol, polyvinyl acetate, polyvinyl alcohol, polyvinyl acetate,RS、RL、RS30D、RL30D、NE30D、K100M、K15M and any combination thereof。

20. The method of claim 15, wherein the coating further comprises a plasticizer.

21. The method of claim 15, wherein the coating further comprises a glidant.

22. The method of claim 21, wherein the glidant comprises talc.

23. The method of any one of claims 11-14, wherein the immediate release particles comprise a seal coating.

24. The method of any one of claims 11-14, wherein the delayed release particles and pulsed release particles comprise a seal coat.

25. The method of claim 13 wherein the first population of pulse-released particles releases the SASGRM after a lag period of about 0.5-3 hours after the SASGRM is released from the immediate-release particles, and the second population of pulse-released particles releases the SASGRM after a lag period of about 0.5-3 hours after the SASGRM is released from the first population of pulse-released particles.

26. The method of claim 25 wherein the first population of pulse-released particles releases the SASGRM after a lag period of about 1-2 hours after the SASGRM is released from the immediate-release particles, and the second population of pulse-released particles releases the SASGRM after a lag period of about 1-2 hours after the SASGRM is released from the first population of pulse-released particles.

27. The method of claim 14, wherein the third population of pulse-released particles releases the SASGRM after a lag period of about 1-3 hours after the SASGRM is released from the second population of pulse-released particles.

28. The method of claim 27, wherein the third population of pulse-released particles releases the sgrm after a lag period of about 1-2 hours after the sgrm is released from the second population of pulse-released particles.

29. The method of any one of claims 11-14, wherein the plurality of particles are contained in a tablet.

30. The method of any one of claims 11-14, wherein the plurality of particles are contained in a capsule.

31. The method of any one of claims 1-9, wherein the subject is a human having symptoms, disorders, or diseases associated with acute stress or transient hypercortisolism.

32. A composition comprising a plurality of particles, the plurality of particles comprising immediate release particles, delayed release particles, or pulsed release particles comprising an amount of a SASGRM.

33. The composition of claim 32, wherein the SASGRM is selected from the group consisting of hydroxy-androst-4, 9(11) -dien-3-one, 21-hydroxy-6, 19-progesterone oxide, 16-hydroxy-11- (substituted phenyl) -estra-4, 9-diene, 17- β -carboxamide for dexamethasone, and Δ 1-11-oxa-11-deoxycorticosterol.

34. The composition of claim 32, wherein the SASGRM is selected from the following compounds:

formula I

Formula II

Formula III

Formula IV

And

formula V

Or a combination thereof and pharmaceutically acceptable salts and solvates thereof.

35. The composition of claim 34, wherein the plurality of particles comprises pulsed release particles.

36. The composition of claim 35, wherein the pulse-release particles comprise a first population of pulse-release particles that release the sgrm after a lag period following release of the sgrm from the immediate-release particles, and a second population of pulse-release particles that release the sgrm after a lag period following release of the sgrm from the first population of pulse-release particles.

37. The composition of claim 36, wherein the pulsed release particles further comprise a third population of pulsed release particles that release the SASGRM after a lag period following release of the SASGRM from the second population of pulsed release particles.

38. The composition of any one of claims 32-37, wherein the delayed-release particles and pulsed-release particles comprise a coating.

39. The composition of claim 38, wherein the coating comprises a water soluble polymer.

40. The composition of claim 39, wherein the water soluble polymer is selected from the group consisting of methylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, cellulose acetate phthalate, polyvinylpyrrolidone/vinyl acetate copolymer, polyvinyl alcohol, polyethylene glycol, polyethylene oxide, hyaluronic acid, alginate, carrageenan, gelatin, and any combination thereof.

41. The composition of claim 38, wherein the coating comprises a water-insoluble polymer.

42. The composition of claim 41, wherein the water insoluble polymer is selected from the group consisting of polyvinyl acetate, cellulose acetate, methyl cellulose, ethyl cellulose, cellulose acetate butyrate, cellulose acetate propionate, non-crystalline cellulose, polyethylene, polyvinyl alcohol, polyacrylates, methacrylates, polyvinyl acetate butyrate, polyvinyl acetate propionate, polyvinyl acetate butyrate, polyvinyl acetate propionate, polyvinyl acetate butyrate, polyvinyl acetate, polyvinyl alcohol, polyvinyl acetate, polyvinyl alcohol, polyvinyl acetate,RS、RL、RS30D、RL30D、NE30D、K100M、K15M and any combination thereof.

43. The composition of claim 38, wherein the coating further comprises a plasticizer.

44. The composition of claim 38, wherein the coating further comprises a glidant.

45. The composition of claim 44, wherein the glidant comprises talc.

46. The composition of any one of claims 32-37, wherein the immediate release particles comprise a seal coating.

47. The composition of any one of claims 32-27, wherein the delayed-release particles and pulsed-release particles comprise a seal coating.

48. The composition of any one of claims 36-37, wherein the first population of pulse-released particles releases the SASGRM after a lag period of about 1-3 hours after the SASGRM is released from the immediate-release particles, and the second population of pulse-released particles releases the SASGRM after a lag period of about 1-3 hours after the SASGRM is released from the first population of pulse-released particles.

49. The composition of claim 48, wherein the first population of pulse-release particles releases the SASGRM after a lag period of about 1-2 hours after the SASGRM is released from the immediate-release particles, and the second population of pulse-release particles releases the SASGRM after a lag period of about 1-2 hours after the SASGRM is released from the first population of pulse-release particles.

50. The composition of claim 37, wherein the third population of pulse-released particles releases the SASGRM after a lag period of about 1-3 hours after the SASGRM is released from the second population of pulse-released particles.

51. The composition of claim 50, wherein the third population of pulse-released particles releases the SASGRM after a lag period of about 1-2 hours after the SASGRM is released from the second population of pulse-released particles.

52. The composition of any one of claims 32-37, wherein the plurality of particles are contained in a tablet.

53. The composition of any one of claims 32-37, wherein the plurality of particles are contained in a capsule.

54. The composition of any one of claims 32-37, wherein the composition comprises the pulsed release particles in an amount effective to prevent, alleviate or treat symptoms, disorders and diseases associated with acute stress or transient hypercortisolism.

55. The composition of any one of claims 32-37, comprising a compound of formula I

Or a pharmaceutically acceptable salt or solvate thereof.

56. The composition of any one of claims 32-37, comprising a compound of formula II

Or a pharmaceutically acceptable salt or solvate thereof.

57. The composition of any one of claims 32-37, comprising a compound of formula III

Or a pharmaceutically acceptable salt or solvate thereof.

58. The composition of any one of claims 32-37, comprising a compound of formula IV

Or a pharmaceutically acceptable salt or solvate thereof.

59. The composition of any one of claims 32-37, comprising a compound of formula V

Or a pharmaceutically acceptable salt or solvate thereof.

Technical Field

The present invention relates generally to the field of preventing, alleviating and treating symptoms, disorders and diseases associated with acute stress or transient hypercortisolism. More particularly, the present invention relates to the prevention, alleviation and treatment of the symptoms of stress-related academic and educational problems, accommodation disorders, insomnia disorders and circadian rhythm disorders by administering immediate release, delayed release and/or pulsatile release compositions of short acting selective glucocorticoid receptor modulators ("SASGRM") including, but not limited to, hydroxy-androst-4, 9(11) -dien-3-one (e.g., RU-43044), 21-hydroxy-6, 19-oxytrogesterone, 16-hydroxy-11- (substituted phenyl) -estra-4, 9-diene (e.g., ORG 36410), 17- β -carboxamide to dexamethasone, and Δ 1-11-oxa-11-deoxycorticosterol.

Background

Throughout this specification, various publications are referenced, including patents, published applications, technical articles and academic papers. Each of these cited publications is incorporated by reference herein in its entirety for all purposes.

All organisms must maintain homeostasis or homeostasis. Stress occurs when homeostasis is disturbed, or is considered to be disturbed, triggering an adaptive response to reestablish homeostasis.¹One of the major peripheral effectors of the stress system is the glucocorticoid, the most important glucocorticoid in humans is cortisol.²

Cortisol is a pleiotropic steroid hormone synthesized and secreted by the adrenal gland. It is one of the major mediators of stress and also of the sleep-wake cycle and acts on many body systems and organs including the cardiovascular system, energy metabolism, immunity and the brain, where cortisol levels affect cognition, memory, the sleep-wake system and other brain functions and neurotransmitters.⁴

Cortisol exerts its effects by binding to the glucocorticoid receptors ("GR") and the mineralocorticoid receptors ("MR") which are two members of the nuclear hormone superfamily. The GR functions as a ligand-activated transcription factor, via the various nonvariable regionsThe homo-mechanism activates or represses transcription of a target gene, including binding to GR response elements as a homo-or heterodimer, by protein-protein interaction with other transcription factors, or by sequestering other transcription factors to inhibit their binding to DNA⁵。

Cortisol levels have a circadian rhythm driven by a pacemaker in the suprachiasmatic nucleus of the hypothalamus, with daily peaks occurring near early morning sleep-wake transitions and lowest levels occurring in the evening and early midnight. In addition, cortisol levels are negatively fed-back regulated through the hypothalamic-pituitary-adrenal (PA) axis, where cortisol inhibition activates the hormones it synthesizes and secretes, namely Corticotropin Releasing Hormone (CRH) in the hypothalamus and adrenocorticotropic hormone (ACTH) in the pituitary gland.⁴In addition to the circadian rhythm of cortisol, the feedback loop also causes a sub-circadian pattern of cortisol secretion with a cortisol peak pulse almost every hour.⁶This pulse rhythm is important for the biological consequences of cortisol signaling. Finally, the enzymes 11- β -hydroxysteroid dehydrogenase (11 β HSD) type 1 and 2 modulate local cortisol concentrations and depend on whether the organ affects MR, GR or both are occupied by cortisol.⁷

Thus, the control of cortisol levels and the mode of action thereof is very complex, and the outcome of its binding to GR depends on the local cortisol concentration, its pulsatility, the level of GR expression, the co-expression of MR, and the presence of tissue and cell specific co-factors and co-repressors at the molecular level. This complexity greatly hinders the recognition of selective SASGREM that is active in vivo and shows a complete decoupling between the desired activity in a variety of different organs or biological systems and the metabolic and immunosuppressive effects on the HPA axis.

Pain that is not proportional to the expected response to the stimulus is defined as an accommodation disorder.⁸The symptoms are clinically significant, resulting in significant distress and dysfunction. The suffering and disorder are associated with stressors.

One of the most common accommodative hurdles with dysfunctions under stress is a discipline disruption. For example, many students do notThe laws deal with stress caused by learning workload or upcoming examinations. In a valid experimental setup of academic and Social pressures (Trier Social Stress Test), cortisol has been shown to rise in this case.⁹The timing of cortisol elevation can impair memory consolidation and memory retrieval, leading to disruption of the industry in addition to acute anxiety.¹⁰

Excessive arousals from a stressed day can severely delay the onset of sleep and disrupt sleep maintenance. The stress-related insomnia causes a decrease in performance and links stress and insomnia together, forming a vicious circle. Sleep is regulated by the interaction of two oscillatory processes, sleep homeostasis and a circadian pacemaker.¹¹The circadian sleep-wake cycle is regulated by a circadian pacemaker located in the anterior suprachiasmatic nucleus of the hypothalamus, whose timing is synchronized (accompanied) with light/dark and social activity. Sleep stages are closely associated with a pattern of pericentral cortisol secretion that reflects activity of the hypothalamic-pituitary-adrenal (HPA) axis. Cortisol peaks shortly after arousal (cortisol arousal response) and then declines during the day to reach a minimum around midnight (nadir). Cortisol changes coincide with sleep cycles where cortisol levels are low in the first half of the night when Slow Wave Sleep (SWS) is dominant and cortisol increases in the second half of the night when Rapid Eye Movement (REM) sleep is dominant.⁴

Certain sleep disorders are associated with hypercortisolism, disorders in the timing and amplitude of cortisol secretion, and hyperactivity of the HPA axis. It has been shown that the insomnia phenotype with subjective chronic primary insomnia and objective short sleep duration is associated with elevated cortisol plasma levels, in contrast to the phenotype with normal objective sleep duration.¹²

Changes in sleep patterns in healthy aging include decreased SWS, increased number of early awakenings and arousals, and have been associated with HPA axis hyperactivity. It was also found that plasma levels of cortisol increase significantly following a cross-time zone trip or experimental sudden time shifts, with profound changes in the temporal pattern of cortisol secretion.¹³It has been found that long-term destruction of the circadian rhythmIncreased salivary cortisol is associated with temporal lobe atrophy.¹⁴

Disclosure of Invention

The present invention describes methods of alleviating, preventing and treating symptoms, disorders and diseases associated with acute stress or transient hypercortisolism comprising administering to a subject in need thereof a short acting selective glucocorticoid receptor modulator ("SASGRM"), including but not limited to SASGRM compounds selected from the group consisting of:

hydroxy-androst-4, 9(11) -dien-3-ones, e.g. compounds of formula I RU-43044

21-hydroxy-6, 19-progesterone oxide, e.g. compounds of formula II

16-hydroxy-11- (substituted phenyl) -estra-4, 9-dienes, e.g. the compound of formula III ORG 36410of

17-beta-carboxamides of dexamethasone, e.g. of formula IV

And

delta 1-11-oxa-11-deoxycorticosterol, e.g. of formula V

Or a combination thereof and pharmaceutically acceptable salts and solvates thereof, in an amount effective to prevent, alleviate or treat symptoms, disorders and diseases associated with acute stress or transient hypercortisolism. States of acute stress or transient hypercortisolism include academic and educational problems associated with stress, adaptation disorders, insomnia disorders, and circadian rhythm disorders. In particular, distress disproportionate to the expected response to a stimulus is defined as a maladaptive disorder. The symptoms are clinically significant, resulting in significant distress and dysfunction. The suffering and disorder are associated with stressors.

In particular, insomnia disorders associated with transient hypercortisolism include stress-related acute insomnia and some forms of chronic insomnia including chronic insomnia with objectively short sleep duration. Circadian rhythm disorders include time-lag type and shift type circadian rhythm disorders.

The invention also describes a composition comprising a plurality of particles, including immediate release particles and/or delayed release particles, which comprise an amount of the SASGRM together with their pharmaceutically acceptable carrier and optionally other therapeutic agents. The SASGRM of the compositions include, but are not limited to: hydroxy-androst-4, 9(11) -dien-3-ones, such as compounds of formula I; 21-hydroxy-6, 19-progesterone oxide, such as a compound of formula II; 16-hydroxy-11- (substituted phenyl) -estra-4, 9-dienes, for example a compound of formula III; 17-beta-carboxamides of dexamethasone, such as the compounds of formula IV; and Δ 1-11-oxa-11-deoxycorticosterol, such as a compound of formula V; and pharmaceutically acceptable salts and solvates thereof, and any combination thereof. The compositions may preferably comprise, for example, immediate release and/or delayed release particles or combinations thereof comprising a SASGRM of formula I, formula II, formula III, formula IV or formula V and pharmaceutically acceptable salts and solvates thereof and any combination thereof, and a pharmaceutically acceptable carrier therefor and optionally other therapeutic agents. The delayed release particles may comprise pulsed release particles.

In certain instances, the composition comprises a population comprising immediate release particles, and first and second populations comprising pulsed release particles. The first population of pulse-release particles releases the SASGRM after a lag period following release of the SASGRM from the immediate-release particles, and the second population of pulse-release particles releases the SASGRM after a lag period following release of the SASGRM from the first population of pulse-release particles. In certain instances, the composition further comprises a third population of pulse-release particles that release the sgrm after a lag period of release of the sgrm from the second population of pulse-release particles. Each lag phase may range from about 0.5 hours to about 3 hours, preferably from about 0.5 hours to about 2.5 hours, more preferably from about 1 hour to about 2 hours.

Also described are compositions that elicit a delayed or pulsatile release profile comprising each of the SASGRMs of formula I, formula II, formula III, formula IV or formula V and pharmaceutically acceptable salts and solvates thereof, and any combination thereof, and a pharmaceutically acceptable carrier therefor and optionally other therapeutic agents. The compositions include an immediate release population comprising the sgrm particles and one or more delayed release or pulsed release populations comprising the sgrm particles, and pharmaceutically acceptable salts and solvates thereof, and any combination thereof, and pharmaceutically acceptable carriers thereof and optionally other therapeutic agents. The SASGRM in each particle may comprise formula I, formula II, formula III, formula IV, or formula V, and pharmaceutically acceptable salts and solvates thereof, or any combination thereof. The compositions include an amount of the SASGRM effective to prevent, alleviate or treat symptoms, disorders and diseases associated with acute stress or transient hypercortisolism.

The first population of pulse-release particles releases the SASGRM after a lag period following release of the SASGRM from the immediate-release particles. In certain instances, the second pulse-released population of particles releases the sgrm after a lag period following the release of the sgrm from the first pulse-released population of particles. In certain instances, the third pulse-released population of particles releases the sgrm after a lag period of time following the release of the sgrm from the second pulse-released population of particles. Each lag phase may be in the range of about 0.5 hours to about 24 hours, preferably 1 hour to about 18 hours. Each lag phase may be in the range of about 1 hour to about 6 hours, preferably about 1 hour to about 5 hours. Each lag phase may be in the range of about 0.5 hours to about 3 hours. Each lag phase may be in the range of about 0.5 hours to about 2.5 hours. Each lag phase may be in the range of about 1 hour to about 2 hours. Thus, the composition may comprise one, two, three or more populations of sgrm pulsed-release particles.

The immediate release, delayed release and/or pulsed release particles may comprise a coating. The coating may comprise a water soluble polymer. The water-soluble polymer may comprise methylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methacrylic acid copolymer, cellulose acetate phthalate, polyvinylpyrrolidone/vinyl acetate copolymer, poly (vinyl acetate),L、S, polyvinyl alcohol, polyethylene glycol, polyethylene oxide, hyaluronic acid, alginate, carrageenan, gelatin, or any combination thereof. The coating may comprise a water insoluble polymer. The water insoluble polymer may comprise polyvinyl acetate, cellulose acetate, methyl cellulose, ethyl cellulose, cellulose acetate butyrate, cellulose acetate propionate, non-crystalline cellulose, polyethylene, chitosan, polyvinyl alcohol, polyacrylates, methacrylates, polyvinyl acetate butyrate, polyvinyl acetate propionate, polyvinyl acetate butyrate, polyvinyl acetate, and polyvinyl acetate,RS、RL、RS30D、RL30D、NE30D、K100M、K15M or any combination thereof.

The coating may optionally include a plasticizer. The plasticizer may comprise triethyl citrate. The coating may optionally include a glidant. The glidant may comprise talc. The coating may optionally include an osmotic agent.

The immediate release, delayed release and/or pulsed release particles may comprise a seal coat.

The immediate release, delayed release and pulsed release particles may be included in a dosage form comprising all particle types, including all delayed release and/or all pulsed release populations. The immediate, delayed release and pulsed release particles may be in separate dosage forms. Each delayed release and/or pulsed release population may be located in a separate dosage form. The dosage form may comprise a tablet, caplet (caplet) or capsule, for example a gelatin capsule.

Drawings

Figure 1 is a graph of the results from example 2, experiment 1 of table 2, for the number of mouse entries;

FIG. 2 is a graph of the results from example 2, experiment 1 of Table 2, for the time spent in the open area;

FIG. 3 is a graph of the results from example 2, experiment 2 of Table 4, for the number of mouse entries;

FIG. 4 is a graph of the results of example 2 experiment 4 from Table 4 for the time spent in the open area;

FIG. 5 is a graph of the results from example 2, experiment 3 of Table 6, for mouse entry times; and is

Figure 6 is a graph of the results from example 2, experiment 3 of table 6, for the time spent in the open area.

Detailed Description

Various terms relating to various aspects of the present invention are used throughout the specification and claims. Unless otherwise indicated, such terms are to be given their ordinary meaning in the art. Other specifically defined terms should be construed in a manner consistent with the definitions provided herein.

As used herein, no specific number of an indication includes a plural indication unless the context clearly dictates otherwise.

The term "immediate release" is known in the art and includes compositions and active ingredients that are formulated (preferably into oral dosage forms) so as to release the composition or active ingredient immediately or as quickly as possible after administration without delaying or prolonging the dissolution or absorption of the composition or active ingredient.

The term "delayed release" is known in the art and includes compositions and active ingredients that are formulated, preferably as oral dosage forms, to release only at some point after initial administration of the composition or active ingredient, rather than immediately after administration.

The term "delayed release" is a form of delayed release known in the art and includes compositions and active ingredients that are formulated, preferably as oral dosage forms, for rapid and immediate release of the composition or active ingredient within a short period of time immediately after a predetermined period of non-release, i.e., lag time, or by a mechanism that rapidly and completely delivers the composition or active ingredient after a lag time, i.e., a period of time without release of the composition or active ingredient. The term also includes controlled release dosage forms, preferably oral dosage forms, having a pulsatile profile which produces at least one timed pulse following an initial immediate release pulse, each pulse being characterized by rapid and substantially complete release of the active ingredient. Such systems are known in the art as pulsatile compound delivery systems (PDDS), time-controlled systems, or S-type release systems, and include commercially available systems such as pulsencap^TM、 And

hydroxy-androst-4, 9(11) -dien-3-one comprises RU-43044((10R,13S,17S) -17-hydroxy-13-methyl-10- [ (4-methylphenyl) methyl ] -17-prop-1-ynyl-2, 6,7,8,12,14,15, 16-octahydro-1H-cyclopenta [ a ] phenanthren-3-one, also known as 17- β -hydroxy-17- α -19- (4-methylphenyl) androst-4, 9(11) -dien-3-one) of formula I:

21-hydroxy-6, 19-progesterone oxide comprises a compound of formula II:

the 16-hydroxy-11- (substituted phenyl) -estra-4, 9-diene comprises ORG 36410((11- β,16- α,17- β) -11- (4-isopropylphenyl) -16, 17-dihydroxy-17- (1-propynyl) estra-4, 9-dien-3-one) of formula III:

the 17- β -carboxamide of dexamethasone comprises a compound of formula IV:

and is

Δ 1-11-oxa-11-deoxycorticosterol comprises a compound of formula V:

the terms subject and patient are used interchangeably. The subject can be any organism, including mammals such as farm animals (e.g., horses, cows, sheep, pigs), laboratory animals (e.g., mice, rats, rabbits), companion animals (e.g., dogs, cats), and non-human primates (e.g., new world monkeys and old world monkeys). Preferably, the subject is a human.

Symptoms and diseases associated with acute stress or transient hypercortisolism include academic and educational problems associated with stress, adaptation disorders, insomnia disorders, and circadian rhythm disorders. Non-limiting examples of academic and educational questions are school exam failures, scores or ratings of failures received or underperformance (lower than expected based on the mental capacity of the individual). Non-limiting examples of accommodation disorders include accommodation disorders accompanied by anxiety characterized by stress, anxiety, nervousness, or dissociative anxiety. Non-limiting examples of insomnia and circadian rhythm disorders include short-term sleep disturbances caused by time zone changes (e.g., jet lag) and shift work, as well as sleep disturbances caused by stressful events (e.g., family, financial or work related distress) and general changes in normal routine sleep and/or environment, as well as certain forms of chronic insomnia, such as chronic insomnia associated with transient hypercortisolism with objective short sleep duration.

The SASGRM of the invention, including hydroxy-androst-4, 9(11) -dien-3-one (e.g., formula I), 21-hydroxy-6, 19-oxytrogesterone (e.g., formula II), 16-hydroxy-11- (substituted phenyl) -estra-4, 9-diene (e.g., formula III), 17- β -carboxamide with dexamethasone (e.g., formula IV), and Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula V), when administered as a continuous, appropriately timed and customized agent, prevents, alleviates, treats, and/or beneficially affects the symptoms, disorders, and diseases associated with acute stress or transient hypercortisolism. The SASGRM of the invention, including hydroxy-androst-4, 9(11) -dien-3-one (e.g., formula I), 21-hydroxy-6, 19-progesterone oxide (e.g., formula II), 16-hydroxy-11- (substituted phenyl) -estra-4, 9-diene (e.g., formula III), Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula IV) and Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula V), has limited metabolic stability, crosses the blood-brain barrier, has selective antagonistic activity at the glucocorticoid receptor, and does not substantially alter cortisol levels in the blood after oral administration (i.e., does not disrupt the HPA axis), which properties can be advantageously exploited in beneficially affecting single-or intermittent multi-dose based regimens, are ideal for preventing, alleviating and treating symptoms, disorders and diseases associated with acute stress or transient hypercortisolism. Accordingly, the invention describes the use of the SASGRMs of the invention including hydroxy-androst-4, 9(11) -dien-3-one (e.g., formula I), 21-hydroxy-6, 19-oxytrogesterone (e.g., formula II), 16-hydroxy-11- (substituted phenyl) -estra-4, 9-diene (e.g., formula III), Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula IV) and Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula V) or any combination thereof and pharmaceutically acceptable salts and solvates thereof and such SASGRMs include hydroxy-androst-4, 9(11) -dien-3-one (e.g., formula I), 21-hydroxy-6, 19-oxytrogesterone (e.g., formula II), A method of ameliorating, preventing and treating symptoms, disorders and diseases associated with acute stress or transient hypercortisolism comprising administering to a subject in need thereof a composition of 16-hydroxy-11- (substituted phenyl) -estra-4, 9-diene (e.g., formula III), Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula IV), and Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula V), or any combination thereof, and pharmaceutically acceptable salts and solvates thereof, and the use of such compositions for preventing, ameliorating and treating symptoms and diseases associated with acute stress or transient hypercortisolism. Preferably, the composition is a pharmaceutical composition comprising delayed release particles and/or pulsed release particles of the SASGRM.

The invention also provides the use of the SASGRM of the invention, including hydroxy-androst-4, 9(11) -dien-3-one (e.g., formula I), 21-hydroxy-6, 19-oxytrogesterone (e.g., formula II), 16-hydroxy-11- (substituted phenyl) -estra-4, 9-diene (e.g., formula III), Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula IV), and Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula V), or any combination thereof, and pharmaceutically acceptable salts and solvates thereof, for the manufacture of a medicament for the alleviation, prevention and treatment of symptoms, disorders and diseases associated with acute stress or transient hypercortisolism. Preferably, the drug is a drug comprising delayed release particles and/or pulsed release particles of the SASGRM.

In general, methods for alleviating, preventing and treating symptoms, disorders and diseases associated with acute stress or transient hypercortisolism include administering to a subject in need thereof an effective amount of a SASGRM comprising hydroxy-androst-4, 9(11) -dien-3-one (e.g., formula I), 21-hydroxy-6, 19-progesterone oxide (e.g., formula II), 16-hydroxy-11- (substituted phenyl) -estra-4, 9-diene (e.g., formula III), Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula IV) and Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula V) or any combination thereof and pharmaceutically acceptable salts and solvates thereof as a therapeutic agent for combating, preventing, alleviating or treating symptoms associated with acute stress or transient hypercortisolism, Part of the protocol for disorders and diseases. Other SASGRMs that have potent metabolic stability, cross the blood brain barrier, have selective antagonist activity at the glucocorticoid receptor, and do not substantially increase cortisol levels in the blood may also be used as part of a regimen to combat, prevent, alleviate or treat symptoms, disorders and diseases associated with acute stress or transient hypercortisolism.

The amount of the SASGRM of the present invention needed to combat, prevent, alleviate or treat the symptoms, disorders and diseases associated with acute stress or transient hypercortisolism, including hydroxy-androsta-4, 9(11) -dien-3-one (e.g., formula I), 21-hydroxy-6, 19-progesterone oxide (e.g., formula II), 16-hydroxy-11- (substituted phenyl) -estra-4, 9-diene (e.g., formula III), Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula IV) and Δ 1-11-oxa-11-deoxycorticosterol (e.g., formula V) or any combination thereof and pharmaceutically acceptable salts and solvates thereof will, of course, vary with the particular compound, route of administration, age and condition of the recipient and the combat, stress, or transient hypercortisolism, Preventing, alleviating or treating a particular symptom, disorder or disease.

A daily dose suitable for any of the above symptoms, disorders or diseases will be in the range of 0.01 to 100mg/kg body weight of the recipient (e.g. human) per day, preferably in the range of 0.01 to 30mg/kg body weight per day, most preferably in the range of 0.1 to 30mg/kg, 0.1 to 20 mg/kg, 1 to 30mg/kg, 1 to 10mg/kg, 1 to 3mg/kg body weight per day. The dose may be 1, 3, 10, 30 or 100mg/kg body weight/day. The desired dose may be provided as 1, 2,3, 4,5 or more divided doses administered at suitable intervals throughout the day. Most preferably, the medicament is provided immediately prior to the recipient sleeping.

The SASGRM or combination thereof may be, for example, formulated as a composition, preferably as a pharmaceutical composition, in immediate release, delayed release and/or pulsed release particles such as described or exemplified herein. The SASGRM or combination thereof is preferably administered in a manner such that it can cross the blood brain barrier of the subject and antagonize glucocorticoid receptors in brain cells to enhance academic performance and adaptation to stress situations, and/or can induce and/or maintain sleep following an acute stress episode or cross-time zone travel or shift work. The subject is preferably a human being and preferably is in need of prevention, alleviation or treatment of symptoms of acute stress in a academic or educational setting due to examination, public speaking or similar social situations requiring adjustments to stress, as well as prevention, alleviation or treatment of insomnia and circadian rhythm disturbances associated with transient hypercortisolism.

Since the SASGRM having selective antagonistic activity at the glucocorticoid receptor is intended to target cells of the brain, the agent is preferably free to pass the blood brain barrier. The SASGRMs are preferably administered according to any method or route suitable to allow the SASGRMs to reach their target cells. Administration may be passive or directed. Administration is preferably oral. In certain instances, administration can be non-oral, e.g., rectal, vaginal, intranasal, topical (including transdermal, buccal, and sublingual), or parenteral administration of the SASGRM. The compositions may be prepared by any method known in the art of pharmacy, for example as described in the examples herein, or using methods such as described in the latest edition of Remington's pharmaceuticals (see, inter alia, Pharmaceutical preparations and their preparation).

Preferably, the SASGRM is delivered in a suitably delayed release or timed pulse, alone or in combination with the immediate delivery of one or more SASGRMs. When delivered in a timed pulse fashion, the short half-life of the SASGRM results in a true pulse of pharmacological activity, rather than sustained pharmacological activity such as seen with mifepristone. The individual pulses of pharmacological activity of the SASGRM can be synchronized with the sub-diurnal pattern of cortisol levels found in healthy subjects, thereby minimizing the derepression of the hypothalamo-pituitary-adrenal (HPA) axis by cortisol. Furthermore, the circadian rhythm of cortisol secretion, in particular the morning peak cortisol known as the wake response, is maintained, thus preserving the subject's ability to adequately adapt to normal stress.

Preferably, the release profile of the SASGRM for alleviation, prevention and treatment of symptoms, disorders and diseases associated with acute stress or transient hypercortisolism comprises a single release from an immediate release composition, preferably in combination with one or more delayed or pulsed releases over a period of several hours, said delayed or pulsed release starting with an immediate release followed by one or more delayed or pulsed releases, with a lag time between each release of a total duration of 0.5-24 hours, preferably 0.5-18 hours, more preferably 0.5-5 hours, more preferably 0.5-3 hours. The delayed or pulsed release is preferably after sleep onset. The SASGRM pulsed compound delivery system may comprise any suitable number of pulses, including 2,3, 4, or 5 pulses. Including the immediate release (pulse 1), the pulsed compound delivery system may further include a second pulse after a lag period following the first pulse, and optionally a third pulse after a lag period following the second pulse, and optionally a fourth pulse after a lag period following the third pulse. In certain preferred aspects, the pulsed compound delivery system comprises a total of two pulses. In certain preferred aspects, the pulsed compound delivery system comprises a total of three pulses. In certain preferred aspects, the pulsed compound delivery system comprises a total of four pulses.

The lag phase comprises the time between pulse delivery of the SASGRM from each population of the SASGRM-containing particles. Preferably, the lag period between the pulses is from about 0.5 hours to about 24 hours, preferably from about 0.5 hours to about 5 hours, preferably from 0.5 hours to about 3 hours, more preferably from about 0.5-1 hour, more preferably from about 1-3 hours, more preferably from about 1 hour to about 2 hours. The lag period between the pulses may be about 0.5 hours, 1 hour, about 1.5 hours, about 1.75 hours, about 2 hours, about 2.25 hours, about 2.5 hours, about 2.75 hours, about 3 hours, about 3.5 hours, about 4 hours, about 4.5 hours, about 5 hours, about 5.5 hours, about 6 hours, or about 18 hours.

Thus, the SASGRM is preferably delivered in 2 to 4 different compound-containing fractions comprising a fast/immediate release fraction, a moderate rate release fraction that releases the SASGRM after a lag period, a moderate to slow release fraction that releases the SASGRM after a second lag period, and a slow release fraction that releases the SASGRM after a third lag period. The final release preferably occurs about 5 to about 9 hours after the immediate release, more preferably about 5 hours or about 6 hours after the immediate release. The intermediate and final released doses may be lower than the previously released doses.

On the basis of the particle fraction of the overall compound delivery system, the SASGRM is preferably delivered according to an immediate release system, more preferably according to a delayed or pulsed release system optionally in combination with an immediate release system. For example, 2 to 4 populations of particles containing a SASGRM can be prepared into larger dosage forms, wherein each population of particles is designed to release the SASGRM according to a desired time frame. Thus, the particles comprising the SASGRM may be prepared, for example, as tablets or caplets, or may be prepared as capsules. The delayed or pulsed release system may also comprise a separately administered population of SASGRM-containing particles, such as immediate release tablets, caplets or capsules, and one or more separate delayed or pulsed release tablets, caplets or capsules.

The SASGRM-containing particles can comprise particles, spherulites, microparticles, nanoparticles, minitablets, beads, granules, or mixtures thereof. When formulated in a tablet or caplet delivery form, the particles may be compressed together. When formulated in a capsule delivery form, the particles may be loaded in a capsule. Suitable delivery forms other than tablets, caplets or capsules may be used.

The particles may comprise an inert core, for example a sugar species having a SASGRM coated thereon. In addition to the SASGRM, the particles may comprise one or more excipients and/or one or more coatings. Common excipients are known in the art and include, but are not limited to, dissolution enhancers, disintegrants, lubricants, binders, glidants, taste-masking or flavoring agents, sweeteners, dyes, and other known excipients. The coating may help to maintain the particle form and influence the release rate and/or release location. The coating preferably comprises a water-soluble and/or water-insoluble polymer or copolymer and may be pH-dependent or pH-independent. In certain instances, the particles comprise a sealing layer that may encase the SASGRM-containing core and/or delayed or pulsed release coated particles. For an immediate release population of particles, a sealing layer may be used in place of the coating layer.

Suitable water-soluble polymers include, but are not limited to, methylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, cellulose acetate phthalate, polyvinylpyrrolidone/vinyl acetate copolymer, methacrylic acid copolymers such asL and S, polyvinyl alcohol, polyethylene glycol, polyethylene oxide, hyaluronic acid, alginate, carrageenan, gelatin, and any combination thereof.

Suitable water-insoluble polymers include, but are not limited to, polyvinyl acetate, cellulose acetate, methyl cellulose, ethyl cellulose, cellulose acetate butyrate, cellulose acetateCellulose propionate, noncrystalline cellulose, polyethylene, chitosan, polyvinyl alcohol, polyacrylate, methacrylate, polyvinyl alcohol, and a cellulose acetate, cellulose acetate, cellulose acetate, cellulose acetate, cellulose acetate, cellulose acetate, cellulose acetate, cellulose acetate, cellulose acetate, cellulose,RS, RL, RS30D, RL30D, and NE30D, and othersA brand of polymer, andK100M and K15M and othersBranded polymers, and any combination thereof.

In some cases, a plasticizer may be included in the coating. Suitable plasticizers include, but are not limited to, glycerol, glycerin, triacetin, glycerin sorbitol, acetates, polyethylene glycol (PEG), triethyl citrate, acetyl triethyl citrate, tributyl citrate, diethyl phthalate, dibutyl phthalate, dihexyl phthalate, castor oil, rapeseed oil, olive oil, sesame oil, mono-and di-acetylated glycerides, and any combination thereof.

In some cases, a glidant may be included in the coating. Suitable glidants include, but are not limited to, magnesium stearate, calcium silicate, magnesium silicate, talc, glyceryl monostearate, and any combination thereof.

In some cases, an osmotic agent may be included in the coating. Suitable classes of osmotic agents include, but are not limited to, salts, sugars, and acids. Suitable osmotic agents include, but are not limited to, magnesium chloride, magnesium sulfate, potassium chloride, potassium phosphate, sodium chloride, sodium phosphate, sodium sulfite, sodium citrate, citric acid, malic acid, fumaric acid, tartaric acid, mannitol, sorbitol, erythritol, dextrose, fructose, galactose, and any combination thereof.

The polymeric coating may be added to the SASGRM core to achieve a weight gain of about 1% to about 40% per particle. The polymeric coating may be added to obtain from about 2% to about 20%, from about 2% to about 15%, from about 2% to about 13%, from about 2% to about 12%, from about 2% to about 10%, from about 2% to about 9%, from about 2% to about 8%, from about 2% to about 7%, from about 2% to about 5%, from about 3% to about 30%, from about 3% to about 20%, from about 3% to about 15%, from about 3% to about 13%, from about 3% to about 12%, from about 3% to about 10%, from about 3% to about 9%, from about 3% to about 8%, from about 3% to about 7%, from about 3% to about 6%, from about 3% to about 5%, from about 5% to about 30%, from about 5% to about 25%, from about 5% to about 15%, from about 5% to about 12%, from about 5% to about 10%, from about 6% to about 15%, from about 6% to about 12%, from about 6% to about 10%, from about 8% to about 12%, from about 12%, or from about 2% to about 12%, or from about 3% of each particle, An increase in weight of about 8% to about 10%, about 9% to about 12%, about 9% to about 13%, about 9% to about 11%, about 10% to about 40%, about 10% to about 20%, about 10% to about 15%, about 15% to about 25%, about 15% to about 20%, about 20% to about 30%, about 20% to about 25%, about 25% to about 40%, about 25% to about 30%, or about 30% to about 35%. Particles intended to contain the second, third or fourth pulse release may contain more and more weight gain, with thicker polymer coatings allowing longer release delays.

In certain instances, the immediate release particles comprise a weight gain from the polymer coating of about 2%, about 3%, about 4%, about 5%, or about 6%. The second delayed or pulsed release particles comprise a weight gain from the polymer coating of about 5%, about 6%, about 7%, about 8%, or about 9%. The third and subsequent delayed or pulsed release particles comprise a weight gain from the polymer coating of about 8%, about 9%, about 10%, about 11%, or about 12%.

The following examples are provided to describe the invention in more detail. They are intended to illustrate rather than to limit the invention.

Detailed Description

Example 1

Metabolic stability and blood brain barrier transport properties of formula I, formula II, formula III, formula IV and formula V

The metabolic stability of each of the compounds of formula I-V is unknown or known to be very short (e.g., the in vitro data for formula I, reported half-lives of 0.33 hours in rat liver microsomes and less than 0.2 hours in rat hepatocytes; formula IV, reported to be converted to unidentified metabolites within 30 minutes in rat liver microsomes). This has led to the previous discontinuation of the development of such short acting GR ligands, supporting metabolically more stable compounds.

The metabolic stability of each compound of formula I, formula II, formula III, formula IV and formula V can be investigated. The compounds should have limited metabolic stability in vitro and in vivo, compared to mifepristone (RU-486, a progesterone receptor modulator that also exhibits antiglucocorticoid properties).

To determine metabolic stability, 10of formula I, formula II, formula III, formula IV and formula V in DMSO^-3Diluting the stock solution to 10^-6M, and in pooled human and rat liver microsome preparations in pH 7.4 potassium phosphate buffer, respectively, incubated at 37 ℃ for more than 45 minutes and samples were taken at 0, 5, 15, 30, 45 and 60 minutes. Ketanserin or testosterone was used as a positive control. The reaction was terminated by deproteinization using an organic solvent, and after mixing and centrifugation, the samples were analyzed by liquid chromatography-mass spectrometry (LC-MS)/MS. Formula I, formula II, formula III, formula IV and formula V should exhibit a half-life of 5 minutes to 1 hour in pooled human and rat microsomes, respectively.

The metabolic stability of formula I, formula II, formula III, formula IV and formula V can then be investigated in vivo in mice. Formula I, formula II, formula III, formula IV and formula V were suspended in 0.5% (w/V) carboxymethylcellulose and single oral doses of 30mg/kg and 100mg/kg were administered to male CD-1 mice (28-36g) via oral feeding tubes, with n-3 per group. Mifepristone at a dose of 10mg/kg and vehicle control were used as reference. Multiple blood samples were obtained by tail bleeding at 0, 5, 15, 30 minutes and 1, 2, 4, 8 and 24 hours post-dose, and whole blood was collected in heparin-coated tubes. Plasma was obtained after centrifugation at 2000x g for 10 minutes and samples were subjected to LC-MS/MS. Plasma half-life in mice after oral gavage was between 30 and 90 minutes. Corticosterone was analyzed in different aliquots of the same sample and no significant increase in formula I, formula II, formula III, formula IV and formula V was found up to a dose of 30 mg/kg.

Next, formula I, formula II, formula III, formula IV, and formula V were evaluated for interaction with the Blood Brain Barrier (BBB). As part of this study, binding to rat and human plasma proteins was studied using equilibrium dialysis against phosphate buffered saline containing 5 μ M of the test compound and warfarin (an anticoagulant) as the reference compound at pH 7.4 and 37 ℃. After equilibration, aliquots were removed from the sample (donor) and buffer (recipient) chambers and subjected to LC-MS/MS. Formula I, formula II, formula III, formula IV and formula V show less than 50% plasma protein binding in both species. In contrast, for mifepristone, more than 95% of plasma protein binding in rat and human serum has been reported. The BBB was assessed in vitro using a synbiotic MDCK cell monolayer and a 5 μ M dose of test compound on the apical and basolateral side of the monolayer for 90 minutes at pH 7.4. After LC-MS/MS analysis of the samples, the apparent permeability and the exclusion ratio in both directions were determined. Formula I, formula II, formula III, formula IV and formula V have moderate to high BBB permeation potential.

Example 2

Short acting selective glucocorticoid receptor modulators in mouse models of stress-induced anxiety

Formula I, formula II, formula III, formula IV and formula V constitute chemically diverse SASGRMs with similar biological properties. They show selectivity for Glucocorticoid Receptor (GR) binding, selective GR antagonism in vitro, limited metabolic stability in vitro and in vivo, no increase in vivo of corticosterone levels after oral administration as an indicator of HPA axis disassembly inhibition in mice, low plasma protein binding and moderate to high BBB penetration potential.

Formula III (also referred to as PND-001) was tested as a representative compound in the SASGRM class by the elevated plus maze test (EPM) in a mouse model of stress-induced anxiety (experiment 1). EPM relies on an inherent conflict between the rodent's tendency to explore new environments on the one hand and the pressure of such exploration on the other hand, thereby avoiding the aversive nature of bright open arms. In particular, the anxiety-reducing compounds increase the frequency of entry into and the time spent in the open arms.

Male CD-1 mice, 4 to 5 weeks old, were housed in groups (6 or 7 mice per cage) and maintained in a room with controlled temperature (21-22 ℃) and reverse light dark cycle (12h/12 h; light on: 17: 30-05: 30; light off: 05: 30-17: 30) with food and water available ad libitum.

Formula III and vehicle (5% dimethyl sulfoxide (DMSO) and 95% polyethylene glycol 400(PEG400)) were administered s.c. 24 hours and 6 hours prior to the EPM assay.

Diazepam as a positive control was administered p.o. 60 minutes prior to the EPM assay.

Formula III was tested at doses of 10, 30 and 100 mg/kg. Diazepam was p.o. administered 60 minutes prior to the EPM test and tested at a dose of 1 mg/kg. (see Table 1).

Table 1: treatment planning

Experimental procedure:

mice were randomly assigned to one of the different experimental groups. Each animal was identified by its group name, cage number, experimental series (days) and number written on the tail with permanent ink (from 1 to 12).

The device is a PVC labyrinth covered with perspex, divided into four equal search arms (21 x 8cm), all interconnected by a small platform (8 x 8 cm). The device was placed 59 cm above the ground. Two arms are open and the other two arms are closed with walls (height: 21 cm).

After compound administration, mice were placed on the platform opposite the closed arms. The number of entries and the time spent in each arm were recorded over a 5 minute period. When an animal places 4 paws in an arm, it is considered to enter the arm.

The device was cleaned with alcohol (70 °) between each animal. Urine and feces were removed from the maze.

During the trial, the visibility of animal handling and operating personnel is minimized as much as possible.

Calculation and statistical analysis

Analysis of variance (ANOVA) was performed on the resulting data. Fisher's PLSD was used for pairwise comparisons, and p-values ≦ 0.05 were considered significant.

Results

Formula III treated mice showed a significant increase in the number of visits and time spent in open arms (table 2).

As shown in fig. 1, formula III is associated with a significant increase in the number of entries into the open arm. The increase was 103%, 81% and 119% for 10, 30 and 100mg/kg of formula III, respectively, compared to the performance of vehicle-treated mice. The graph of fig. 1 has a mean of n-12 +/-mean standard error for all groups.

As shown in fig. 2, formula III also causes an increase in the time spent in the open arms. The increase was 224%, 154% and 219% for 10, 30 and 100mg/kg of formula III, respectively, compared to vehicle-treated mice. The graph of fig. 2 has a mean of n-12 +/-standard error of the mean for all groups.

Diazepam treatment was associated with a significant increase in both the number of entries and the time spent in the open arms. The increase was 233% and 327% compared to the performance of vehicle-treated mice, respectively.

Table 2: performance during EPM testing

In the next experiment in the elevated plus maze experiment (experiment 2: performed as described under the same experimental procedure and analysis as described in experiment 1), the efficacy of formula III (also referred to as PND001) at lower doses was investigated. In addition, the potency of formula IV (also known as PND002) was studied at the dose showing the potency of formula III in experiment 1. Compound or vehicle (5% dimethyl sulfoxide (DMSO) and 95% polyethylene glycol 400(PEG400)) was administered 24 hours and 6 hours s.c. prior to the EPM assay (table 3).

Table 3: treatment planning

Formula III (PND 001) was tested at doses of 1, 3 and 10 mg/kg. Formula IV (PND002) was tested at a dose of 30 mg/kg. Diazepam as a positive control was administered at a dose p.o of 1mg/kg 60 minutes prior to the EPM assay and tested.

As shown in fig. 3 and table 4, PND001 was associated with a significant increase in the number of entries into the open arm. The graph of fig. 3 has a mean value of n-12 +/-SD for all groups. The increases were 50%, 93% and 56% for PND001 at 1, 3 and 10mg/kg, respectively, with two higher doses reaching statistical significance compared to the performance of vehicle-treated mice. As shown in figure 4 and table 4, PND001 also caused an increase in the time spent in the open arm, which was statistically significantly different from vehicle at all doses. The graph of fig. 4 has a mean value of n-12 +/-SD for all groups. The increase was 60%, 112% and 70% for PND001 at 1, 3 and 10mg/kg, respectively, compared to vehicle treated mice. PND002 treatment was associated with a significant increase in both the number of entries and the time spent in the open arm. The increase was 77% and 90% compared to the performance of vehicle-treated mice, respectively. Diazepam treatment as a negative control was associated with a significant increase in both the number of entries and the time spent in the open arms. The increase was 146% and 151% compared to the performance of vehicle-treated mice, respectively.

Table 4: performance during the EPM test (experiment 2)

In the third experiment in the elevated plus maze trial (experiment 3: performed as described under the same experimental procedure and analysis as described in experiment 1), the effect of the route and protocol of administration was investigated. Formula III (also known as PND001) or vehicle (5% dimethyl sulfoxide (DMSO) and 95% polyethylene glycol 400(PEG400)) was administered by gavage (p.o.). Diazepam as a positive control was administered i.p. 30 minutes prior to the EPM assay.

PND001 was tested at a dose of 3mg/kg with the highest efficacy in experiment 2 and was administered twice 6 and 1 hour prior to the EPM test, or once only 1 hour prior to the test. Diazepam was administered and tested at a dose of 1mg/kg i.p. 30 minutes prior to the EPM test (see table 5).

Table 5: treatment plan (experiment 3)

Injecting: group 3 received vehicle dosing 6 hours prior to the trial to achieve a consistent number of doses with groups 1 and 4.

As shown in table 6 and fig. 6, PND001 caused an increase in the time spent in the open arm. The graph of fig. 6 has a mean of n-8 +/-mean standard error for all groups. The increase was 50% and 91% for 1 hour pre-treatment and 1 hour and 6 hour pre-treatment with PND001, respectively, compared to vehicle treated mice. Statistical significance was achieved only before 1 hour and 6 hours. Treatment prior to 1 hour and 6 hours, rather than just 1 hour, showed an increase in the number of entries. As shown in table 6 and figure 5, the number of entries was approximately doubled compared to that seen in experiment 2 using 24 and 6 hour s.c. pre-treatment at the same dose, but not statistically significant compared to vehicle in this experiment. The graph of fig. 5 has a mean of n-8 +/-mean standard error for all groups.

Diazepam treatment as a positive control was associated with a significant increase in both the number of entries and the time spent in the open arms. The increase was 400% and 270%, respectively, compared to the performance of vehicle-treated mice.

Table 6: performance during the EPM test (experiment 3)

Based on the results of formula III and formula IV reflecting adaptation to stress in a mouse anxiety model, compounds of formula I, formula II and formula V formulated into suitable pharmaceutical dosage forms and SASGREM with similar biological properties (specificity for GR, pure GR antagonist activity in CNS, short plasma half-life and BBB penetration) can be used as beneficial agents for preventing, alleviating or treating symptoms, disorders and diseases associated with acute stress or transient hypercortisolism, in particular for modulating (acute) insomnia, hypercortisolism-associated chronic insomnia and circadian rhythm disorders of the time-lag and round-shift type.

Pharmaceutical dosage forms suitable for the targeted use of sascrem, e.g. of formula I, formula II, formula III, formula IV and formula V in stress disorders, in particular sleep disorders associated with cortisol-hyperactivity, include orally administered controlled release dosage forms having a delayed release or pulsatile release profile which produces at least one timed pulse after an initial immediate release pulse, each pulse being characterised by a rapid and substantially complete release of the active ingredient. Such systems are known as delayed release systems, pulsatile compound delivery systems (PDDS), time controlled systems or S-type release systems.

Example 3

Preparation of coated pellets for immediate release (micronized RU 43044 (formula I))

The particle size of RU 43044 (formula I) can be significantly reduced, for example, by using a specific milling technique such as jet milling in a steel chamber under nitrogen pressure. Micronization occurs when RU 43044 (formula I) powder is fed to the milling chamber at a speed of about 50m/s and the faster particles accelerated to a speed of about 300m/s by a series of gas jet nozzles collide with the slower particles of the input. The particle size distribution (by volume) of RU 43044, which can be achieved by the jet milling technique described above, was about 2-3 μm (d50), 10-12 μm (d99) and <15 μm (d100) measured using laser diffraction.

The preferred dosage form is a multiparticulate system consisting of multiparticulates of 1-3mm in size, which ensures rapid gastric emptying, low variability of gastric transit time and optimized compound absorption.

Step 1. preparation of uncoated pellets containing RU 43044 (formula I). To 60 grams of microcrystalline cellulose: (PH-101) and 8 grams of hydroxypropyl methylcellulose (HPMC E6) 30 grams (g) of micronized RU 43044 (formula I) was added. After homogenizing the dry powder, the mixture was granulated by adding a suitable amount of aqueous PVP (25.000MW) (5%, w/w). The resulting wet granules were extruded at 25rpm at room temperature using a CLS extruder 20(2mm orifice diameter) and then the extrudate was spheronized using a CLS MBS 120 spheronizer with friction plates. The resulting spherical pellets were dried to a moisture of 3-5% containing about 30% (w/w) micronized RU 43044 (formula I).

Step 2. preparation of coated pellets for immediate release containing micronized RU 43044 (formula I). Using a shear mixer, 100ml of a spray suspension containing 6.25 grams of amino methacrylate copolymer-NF: (N-methyl acrylate copolymer) dissolved/suspended in acetone/isopropanol 1:1 was preparedE100) 0.625 g polyethylene glycol (PEG 6000) and 3.1 g (g) talc. Finally the spray suspension was passed through a 0.5mm sieve.

The resulting suspension was sprayed onto 50 grams of pellets prepared according to step 1 using a Caleva laboratory coater MCD 2. The process conditions are set to: the spraying speed is 0.2-0.3g/min, the inlet temperature is 40-45 ℃, the flow rate is 60L/min under 2 bar, and the air temperature is 30-35 ℃. The spray was maintained until the weight increased by 5% due to the application of the coating. Finally the pellets are cured at an air temperature of 30 ℃ to 35 ℃ for about 1 hour.

Example 4

Preparation of layered pellets for delayed or pulsed release (of micronized RU 43044 (formula I))

A spray suspension was prepared using a shear mixer containing 5.25 grams of quaternary ammonium methacrylate copolymer(s) (dissolved/suspended in 300 grams of acetone/isopropanol 1:1RL PO A type), 17.25 g of a quaternary ammonium methacrylate copolymer (C: (R)RS PO type B), 3.5 g triethyl citrate (TEC) and 5.25 g talc. Finally the spray suspension was passed through a 0.5mm sieve.

The resulting suspension was sprayed onto 50 grams (g) of pellets prepared according to step 2 (example 3) using a Caleva lab coater, MCD 2. The process conditions are set to: the spraying speed is 0.2-0.3g/min, the inlet temperature is 40-45 ℃, the flow rate is 60L/min under 2 bar, and the air temperature is 30-35 ℃. The spray was maintained until a 7% weight gain was achieved due to the application of the coating. Finally the pellets are cured at an air temperature of 30 ℃ to 35 ℃ for about 1 hour.

The modulating permeability of the coating membrane and the osmotic agent incorporated into the core of the multiparticulate particle are considered. To achieve a suitable osmotic pressure within the coated multiparticulates, a pharmaceutical grade osmogen of hydrophilic nature may be applied. They are preferably low molecular weight sugars such as fructose, sucrose, mannitol, inorganic salts such as sodium phosphate, or organic acids such as citric acid or tartaric acid. Upon penetration of the gastrointestinal fluids through the film of the coated dosage form following oral administration, dissolution of the osmogen creates osmotic pressure which ultimately results in rupture of the polymer membrane and spontaneous and complete release of the compound.

The breakdown of the coating is influenced by the permeability of gastrointestinal fluids, which can be adjusted by adding hydrophilic low or high molecular weight compounds to the coating. Such regulators may be physiologically inert water soluble polymers such as low molecular weight methylcellulose or Hydroxypropylmethylcellulose (HPMC), sugars such as monosaccharides such as fructose and glucose, disaccharides such as lactose, sucrose, or polysaccharides such as cellulose, amylose and dextran.

To achieve suitable timing of the release pulse, the modulator may be at least 10%, at least about 20%, and generally no more than about 50%, preferably no more than about 40% by weight of the multiparticulate. The polymeric coating may comprise at least about 5% and no more than 50% by weight of the multiparticulate.

The exact ratio of modulator to active agent can be determined by formulation experiments that produce different types of multiparticulates containing different amounts of modulator. The USP approved method of dissolution or release testing can be used to measure release rates (<711>, USP 32NF 27,2009, vol.1, device 1 with nominal capacities varying from 1L to 4L). To better simulate the conditions present in the digestive system, a USP-compliant flow cell apparatus (apparatus 4) may be used instead to determine the dissolution rate if testing under infinite sink conditions is desired. The detection of dissolved compounds as a function of time can follow various methods of the prior art, such as spectrophotometry, HPLC, mass spectrometry, etc., until the absorbance becomes constant or until more than 90% of the compound has been released.

Example 5

Evaluation of Compound Release characteristics for immediate Release RU 43044 (formula I) coated pellets

The in vitro release rate of RU 43044 (formula I) of pellets made according to example 3, step 2 was determined using a USP apparatus according to the well-described spin basket method (Sotax AT). The dissolution vehicle was 900ml deionized water. The temperature was set and controlled at 37 ℃ during the test. The basket rotation speed was set to 50rpm and kept constant. Unit dose pellets were placed in a drying basket at the start of the test.

To determine the concentration of RU 43044, samples were taken from the dissolution vehicle at 5, 10, 20, 30, 45, 60 and 90 minutes and filtered through a Distek 10 μm PE filter. A conventional HPLC apparatus equipped with a UV spectrometer with an absorption of 245nm was used to measure the concentration of RU 43044 in the sample. There is immediate release of the compound from the dosage form. After 45 minutes, more than 80% of the dose is released, and after 90 minutes, the compound is almost completely released and dissolved in the dissolution vehicle. The amount of compound released and the time profile of release are influenced by the amount of coating, the amount of talc, the curing time and the basket stirring rate.

Example 6

Evaluation of Compound Release characteristics of layered pellets for delayed or pulsed Release RU 43044 (formula I)

At the start of the test, unit doses of the pellets from example 4 were placed in a drying basket. The test conditions with respect to rotation speed, temperature and dissolution vehicle were the same as in example 5. To determine the concentration of RU 43044 (formula I), samples were taken from the dissolution vehicle at 30, 60, 90, 120, 150, 180 and 240 minutes and filtered through a Distek 10 μm PE filter. A conventional HPLC apparatus equipped with a UV spectrometer with an absorption wavelength of 245nm was used to measure the concentration of RU 43044 (formula I) in the sample. There is a delayed release of the compound from the dosage form. After 60(90) minutes, no more than 10% (15%) of the dose is released, after 120 minutes no more than 20% of the compound is released, after 150 minutes no more than 80% of the compound is released, and after 180 minutes the release is almost complete. The amount of compound released and the time profile of release are influenced by the amount of coating, the amount of talc, the curing time and the basket stirring rate.

The invention is not limited to the embodiments described and illustrated above, but may be adapted and modified within the scope of the following claims.

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