Indian Patents. 280053:A PHARMACEUTICAL COMPOSITION COMPRISING 3 BETA HYDROXY 5 ALPHA PREGNAN 20 ONE WITH IMPROVED STORAGE AND SOLUBILITY PROPERTIES

Title of Invention	A PHARMACEUTICAL COMPOSITION COMPRISING 3 BETA HYDROXY 5 ALPHA PREGNAN 20 ONE WITH IMPROVED STORAGE AND SOLUBILITY PROPERTIES
Abstract	It is provided apharmaceutical composition comprising 3 beta hydroxy 5 alpha pregnan 20 one at least one sterol or an ester thereof and a mixture of acylglycerols with a solid fat content of less than 25% at 25°C and 0% at 37°C.In addition it is provided a method for preparing the pharmaceutical composition.

Full Text	A pharmaceutical composition comprising 3-beta-hydroxy-5-alpha-pregnan-20-one with improved storage and solubility properties Technical field [0001] The present invention relates generally to an improved formulation of 3- beta-hydroxy-5-alpha-pregnan-20-one. Background art [0002] 3-beta-hydroxy-5-alpha-pregnan-20-one is a steroid in the pregnane family and a modulator of GABA,-receptor activity which is indicated for the treatment of sex/stress steroid induced disorders conditions (W099/4593 1 ). 3-beta-hydroxy-5- alpha-pregnan-20-one is poorly soluble in many therapeutically acceptable solvents, which makes it difficult to administer the compound to a patient. [0003] In animal studies, 3-beta-hydroxy-5-alpha-pregnan-20-one has been intravenously administered to rats in a formulation containing cyclodextrin (W099/4593 1 ). [0004] Grant et al (JPET 326:354-362, 2008) has administered 3-beta-hydroxy-5- alpha-pregnan-20-one to monkeys by using a formulation with hydroxypropyl Pcyclodextrin. [0005] Formulations with cyclodextrins are not suitable for administration to human patients. One reason for this is, because 3-beta-hydroxy-5-aIpha-pregnan-20- one is poorly soluble, the formulation results in a large therapeutic volume that can only be administered intravenously. [0006] Since 3-beta-hydroxy-5-alpha-pregnan-20-one is poorly soluble in water there is still no pharmaceutically acceptable formulation for this compound. Definitions [0007] As used in the present application, the following terms have the specified meanings unless otherwise specified. [0008] By "acylglycerol" is meant all types and combinations of fatty acids esterified to glycerol. [0009] By "medium-chain acylglycerol" is meant a mixture of acylglycerols where the total com bined percentage of octanoic (caprylic) acid and decanoic (capric) acid is at least 95%. [00010] By "solid fat content" is meant the percentage of solid as determined by pulse NMR (nuclear magnetic resonance). [0001 11 "Room temperature" denotes a temperature of between 1 8°C and 25°C. [000 1 21 "UC 1 0 1 0"denotes 3-beta-hydroxy-5-alpha-pregnan-20-one. [000 1 31 "Sterol or ester thereof" denotes steroids with at least one hydroxyl group and esters of said steroids where at least one hydroxyl group has been used for the synthesis of an ester. [00014] Steroids, such as sterols, are usually described by the number of carbon atoms in the compound. Thus, for example, cholesterol is a C27 sterol, which indicates that the compound consists of 27 carbon atoms. [000 1 51 Unless stated otherwise, concentrations are stated as mg/g, that is, mg per gram of pharmaceutical composition. Brief description of fig- ures [000 1 61 Figure 1 shows mean plasma concentrations of 3-beta-hydroxy-5-alphapregnan- 20-one (ng/mL) in rabbits following subcutaneous administration of 3-betahydroxy- 5-alpha-pregnan-20-one in sesame oil with cholesterol (1 : 1 ) in two doses: 1 mg/kg (squares) and 5 mg/kg (circles). [000 1 7] Figure 2 shows 3-beta-hydroxy-5-alpha-pregnan-20-onec oncentration in filtrate (0.2 pm) from a suspension of 3-beta-hydroxy-5-alpha-pregnan-20-one in relation to cholesterol:3-beta-hydroxy-5-aIpha-pregnan-20-one ratio. [00018] Figure 3a, 3 b, 4a and 4b show photographs of suspensions of 3-betahydroxy- 5-alpha-pregnan-20-one. Summary of invention [00019] An object of the present invention is to provide an improved formulation of 3-beta-hydroxy-5-alpha-pregnan-20-one in a pharmaceutically acceptable carrier. [00020] Another object of the present invention is to provide a formulation of 3- beta-hydroxy-5-alpha-pregnan-20-onew ith enhanced storage properties. [00021] Yet another object is to provide a formulation of 3-beta-hydroxy-5-alphapregnan- 20-one with improved pharmacokinetics. [00022] Yet another object of the present invention is to provide a formulation of 3- beta-hydroxy-5-alpha-pregnan-20-onew ith increased solubility in a pharmacologically acceptable carrier. [00023] These and other objects are met by a first general aspect of the present invention which provides a pharmaceutical formulation comprising 3-beta-hydroxy-5- alpha-pregnan-20-one, at least one sterol or an ester thereof and a mixture of acylglycerols with a solid fat content of less than about 25% at 25°C and about 0% at 37°C. [00024] In a second general aspect of the present invention there is provided methods for preparing a pharmaceutical composition. [00025] In a third general aspect of the present invention there is provided a pharmaceutical composition obtainable according to a method according to the invention. [00026] In a fourth general aspect of the present invention there is provided use of a pharmaceutical composition for the treatment of conditions of the central nervous system. Detailed description [00027l The inventors have found that the addition of a sterol surprisingly increases the solubility and improves the pharmacokinetics of 3-beta-hydroxy-5-alphapregnan- 20-one in acylglycerols. [0002 81 Generally, the pharmaceutical composition comprises 3-beta-hydroxy-5- alpha-pregnan-20-one, at least one sterol or an ester thereof and a mixture of acylglycerols with a solid fat content of less than about 25% at 25°C and about 0% at [00029] In particular a sterol with a hydroxyl group bound to the third carbon atom of the sterol structure is useful in the invention. The sterol may be cholesterol or beta-sitosterol, but also other sterols such as stigmasterol, brassicasterol or avenasterol may be used. In particular, cholesterol may be used. [00030] In addition, cholesteryl esters can be used. Examples of such esters are sodium cholesteryl sulphate, cholesteryl bensoate, cholesteryl acetate, cholesteryl caprylate, cholesteryl decanoate, cholestyl palmitate, cholesteryl oleate and cholesteryl steara te. [00031] The sterol or ester thereof can be a C18-C30 sterol or an ester thereof, a C21 -C27 sterol or an ester thereof, or a C27-C29 sterol or ester thereof. [00032] The pharmaceutical composition can, in a first embodiment, be such that 3-beta-hydroxy-5-alpha-pregnan-20-one is essentially dissolved in the composition. Thus 3-beta-hydroxy-5-alpha-pregnan-20-onec an be dissolved or essentially dissolved according to this embodiment of the invention. [00033] The weight ratio of sterol (or ester thereof) to 3-beta-hydroxy-5-alphapregnan- 20-one can, in this embodiment, be in the range of about 1 : 10 to 10: 1 . The sterol or ester thereof may be added in an amount that is similar to the amount of 3- beta-hydroxy-5-alpha-pregnan-20-one by weight. Because 3-beta-hydroxy-5-alphapregnan- 20-one and a sterol have similar molecular weights this results in almost equimolar amounts of 3-beta-hydroxy-5-alpha-pregnan-20-oneto sterol. [00034] Thus, the weight ratio of sterol to 3-beta-hydroxy-5-alpha-pregnan-20-one can be in the range of from 1 :5 to 5: 1 . In particular, the weight ratio of sterol to 3- beta-hydroxy-5-alpha pregnan-20-one can be from 1 :3 to 3: 1 . [00035] Suitable concentrations of 3-beta-hydroxy-5-alpha pregnan-20-one are between 0.1 mg/g and 75 mg/g. The concentration of 3-beta-hydroxy-5-alpha pregnan-20-one can also be between 1 mg/g and 50 mg/g, between 5 mg/g and 30 mg/g or between 10 mg/g and 25 mg/g. [00036] Alternatively, in a second embodiment, the pharmaceutical composition comprises a suspension of 3-beta-hydroxy-5-alpha-pregnan-20-one. In this case the pharmaceutical composition will comprise 3-beta-hydroxy-5-alpha-pregnan-20-one in particles as well as 3-beta-hydroxy-5-alpha-pregnan-20-oned issolved in the composition. The sterol increases the soluble fraction of 3-beta-hydroxy-5-alphapregnan- 20-one in such a suspension compared to a suspension without a sterol. One advantage with a suspension is that the formulation can contain a high concentration of 3-beta-hydroxy-5-alpha-pregnan-20-one.A n additional advantage with a composition that comprises a suspension is that it results in slow release of 3-betahydroxy- 5-alpha-pregnan-20-one. [00037] When the pharmaceutical composition comprises a suspension, the particles are preferably of a range of sizes that is not engulfed by macrophages. Macrophages do primarily engulf particles of a size that is 2-3 micrometer (Champion et al, Pharm Res 2008; 25(8): 1 8 15-1 82 1 ). [00038] In this second embodiment, the weight ratio of sterol (or ester thereof) to 3- beta-hydroxy-5-alpha pregnan-20-one can be in the range of about 1 : 1 0 to 1 0: 1 . The weight ratio of sterol to 3-beta-hydroxy-5aIpha pregnan-20-one can be in the range of from 1 :5 to 5: 1 . In particular, the weight ratio of sterol to 3-beta-hydroxy-5-alphapregnan- 20-one can be from 1 :4 to 3:1 or from 1 :3 to 3:1. [00039] Suitable concentrations of 3-beta-hydroxy-5-alpha-pregnan-20-onea re, in this second embodiment, between 0.1 mg/g and 750 mg/g. The concentration of 3- beta-hydroxy-5-alpha-pregnan-20-one can also be between 1 mg/g and 300 mg/g, between 1 mg/g and 100 mg/g, between 1 mg/g and 50 mg/g, between 5 mg/g and 30 mg/g or between 10 mg/g and 25 mg/g. [00040] The following applies to the invention in general. [0004 11 Generally, the mixture of acylglycerols is characterized in that it has a solid fat content of less than about 25% at 25°C and about 0% at 37°C. Thus, the solid fat content is, for practical purposes, 0% at 37°C. The solid fat content is at most 0.01 % at 37°C. [00042] The mixture of acylglycerols can be a vegetable oil. Thus, it can be a vegetable oil selected from the group consisting of sesame oil, peanut oil, olive oil, and castor oil, or mixtures thereof. [00043] In particular the mixture of acylglycerols can be a medium-chain acylglycerol, that is, a mixture of acylglycerols wherein the total combined percentage of fatty acids with 8 carbon atoms (octanoic acid) and 10 carbon atoms (decanoic acid) is at least 95%. The medium-chain acylglycerol can be various mixtures of monoacylglycerols, diacylglycerols and triacylglycerols. [00044] The medium-chain acylglycerol can consist of from about 50% to about 65% of monoacylglycerols, about 25% to about 35% of diacylglycerols, less than about 5% of triacylglycerols and less than about 2.5% of glycerol. An example of such a medium chain acylglycerol is Akoline MCM. [00045] The medium-chain acylglycerol can be such that it comprises at least about 95% triacylglycerols. Akomed R MCT is an example of such a medium-chain acylglycerol. [00046] The mixture of acylglycerols can comprise a mixture of a vegetable oil and a medium-chain acylglycerol. The mixture of acylglycerols can comprises a mixture of castor oil and a medium-chain acylglycerol where castor oil is present in an amount of between 40% and 60% by weight. The mixture of acylglycerols can consist of about 48% by weight of castor oil and about 52% by weight of a medium-chain acylglycerol. In particular the mixture of acylglycerols can consist of about 48% by weight castor oil and about 52% by weight of a medium-chain acylglycerol. [00047] The pharmaceutical composition may comprise additional excipients known to a person skilled in the art such as antioxidants, preservatives, surfactants, coloring, flavoring, or thickening agents. [00048] The pharmaceutical composition can be administered to the patient by different means. Thus, it may be administered orally, parenterally or topically. Thus, the pharmacological composition may be administered subcutaneously, intramuscularly, intravenously, nasally, transdermally or vaginally. [00049] In a second general aspect of the present invention there is provided methods for preparing a pharmaceutical composition of 3-beta-hydroxy-5-alphapregnan- 20-one. [00050] One method, in which 3-beta-hydroxy-5-alpha-pregnan-20-one is dissolved or essentially dissolved in the composition, comprises the steps of a) dissolving 3-betahydroxy- 5-alpha-pregnan-20-one in ethanol, b) adding a mixture of acylglycerols with a solid fat content of less than about 25% at 25°C and about 0% at 37°C and a sterol or ester thereof, c) mixing until a homogeneous liquid is obtained and d) evaporating the ethanol. [00051] When the mixture of acylglycerol is a solid or a semi-solid at room temperature, such as a medium-chain acylglycerol, the method may comprise a further step, which is the melting of the medium-chain acylglycerol before mixing it with the ethanol-drug preparation. The melting step enables the homogeneous mixing of this type of acylglycerol with other components. Once melted and mixed with the other components, the preparation remains in a liquid state for at least the time periods indicated in Table 1. [00052] When the formulation comprises a suspension, the formulation is advantageously prepared according to a method that comprises the following steps: 1 ) dissolving or suspending the sterol or ester thereof in the mixture of acylglycerols, 2) suspending 3-beta-hydroxy-5-alpha-pregnan-20-one in the acylglycerol-sterol mixture, 3) gently mixing. Surprisingly, this procedure leads to suspended particles comprising 3-beta-hydroxy-5-alpha-pregnan-20-oneo f smaller size. [00053] In a third general aspect of the present invention there is provided pharmaceutical compositions obtainable by the methods according to the second aspect of the invention. [00054] In a fourth general aspect of the present invention there is provided the use of the pharmaceutical composition according to the invention for the treatment or prevention of conditions of the central nervous system. [00055] The pharmaceutical composition can be used to treat or prevent conditions of the central nervous system. Examples of such conditions that can be treated are epilepsy, menstruation cycle dependant epilepsy, depression, stress related depression, migraine, tiredness and in particular stress related tiredness, premenstrual syndrome, premenstrual dysphoric disorder, menstrual cycle linked mood changes, stress related memory changes, menstrual cycle linked memory changes, Alzheimer's dementia, menstrual cycle linked difficulties in concentration, menstrual cycle linked sleep disorders and tiredness, substance abuse, menstrual cycle linked alcoholism, or com binations thereof. [00056] In particular the pharmaceutical composition can be used to treat steroid induced mood disorders, in particular premenstrual dysphoric disorder. [00057] The pharmaceutical composition can also be used for treating or preventing side effects of oral contraceptives and postmenopausal therapy. [00058] The pharmaceutical composition can also be used for control or termination of steroid-induced anesthesia. Examples [00059] The invention will now be described with non-limiting examples. [00060] In order to find a pharmaceutical composition comprising 3-beta-hydroxy- 5-alpha-pregnan-20-one, several different vehicles and com binations of vehicles were evaluated. 3-beta-hydroxy-5-alpha-pregnan-20-onew as dissolved in various vehicles and was evaluated by eye for physical stability at room temperature over time. A formulation that did not undergo any visible changes in appearance and remained clear on storage in room temperature without signs of haziness, precipitation, sedimentation, phase separation into two or more liquid phases or change of colour, within 30 days was considered as "stable". [00061] In addition, the particle size and solubility of 3-beta-hydroxy-5-alphapregnan- 20-one in formulations comprising a suspension of 3-beta-hydroxy-5-alphapregnan- 20-one were evaluated. General procedure for examples 1-49 [00062] The following procedure was adopted for the preparation of 3-betahydroxy- 5-alpha-pregnan-20-one-containing formulations. [00063] The desired amount of 3-beta-hydroxy-5-alpha-pregnan-20-onea nd sterol (for example cholesterol) was weighed in a 100 ml or 250 ml round-bottomed flask. To every gram of the mixture of 3-beta-hydroxy-5-alpha-pregnan-20-one and sterol a volume of about 30 ml of absolute ethanol was added. The mixture was treated in an ultrasonication bath (not exceeding 50°C) until a clear liquid was obtained. This was normally accomplished within 10 minutes. The additional lipid ingredients indicated in the "Vehicle" column in Table 1 was then added up to 20 g. The resulting mixture was gently shaken by hand until a clear, homogeneous liquid was obtained. When the lipid was a solid at room temperature it was melted in the warm ultrasonication bath to a liquid form before addition. [00064] Compounds from the following suppliers were used (product numbers within brackets): Cholesterol from Sigma (C8503), olive oil and peanut oil from Apoteket, Sweden (26 36 16 and 26 66 01, respectively), sesame oil from Fluka (85067), and castor oil from Sigma (259853). Akomed R medium-chain triacylglycerol (MCT) and Akoline medium-chain monoacylglycerol (MCM) were both from AarhusKarlshamns Sweden AB, Karlshamn, Sweden. [00065] The alcohol was evaporated from the liquid on a rotary evaporator at a pressure of about 25 mbar and a temperature of about 40°C until the weight of the flask was essentially constant. The remaining ethanol content was essentially less than 1 %. The aim was to obtain a liquid with the appearance of clear oil at room temperature. The oily liquid was then transferred to clear glass vials and stored at room temperature until evaluation. [00066] The samples were evaluated by observing the samples in the glass vials and recording signs of haziness, precipitation, sedimentation, phase separation into two or more liquid phases, or change of colour after 1 or 2 days after preparation and after 30 days after preparation. In some cases, other time intervals were used (indicated in Table 1). Where indicated, the entire sample was placed in a refrigerator (2-8°C) to provoke precipitation. Example 1 [00067] 3-beta-hydroxy-5-alpha-pregnan-20-one (UC 1 0 1 0) (5 mg/g) and peanut oil was mixed into an emulsion with ethanol as described above in the concentrations shown in Table 1, and the ethanol was evaporated. The final weight of the preparation was 20 g. The mixture then had the form of an oily liquid. After one day when the sample was evaluated there were signs of precipitation. At 30 days the precipitate has formed a bottom sediment. Thus, the formulation was not stable. Example 2 [00068] Example 2 was carried out essentially as Example 1 with the difference that cholesterol (5.5 mg/g) was added. When the sample was evaluated after one day the appearance of the sample had not changed. It was still unchanged after 30 days and after four months. After five months there was a slight precipitation. Example 2 compared with Example 1 shows how the addition of 5.5 mg/g cholesterol to a solution of 5 mg/g 3-beta-hydroxy-5-alpha-pregnan-20-one in peanut oil substantially increases the solubility so that, instead of precipitating, no precipitation occurred and the sample was stable for four months. However, a slight sedimentation occurred after 5 months. Examples 3 to 49 [00069] Examples 3 to 49 were carried out essentially as described above with the variations with regard to 3-beta-hydroxy-5-alpha-pregnan-20-onec oncentration, acylglycerol mixture used, sterol used and sterol concentration that are shown in Table 1. [00070] The data from examples 1 to 49 is presented in Table 1 . The effect of addition of cholesterol is, for example, evident in examples 8 and 12, where the addition of cholesterol (1 0 mg/g) substantially increased solubility so that the sample did not precipitate but instead was stable for 12 months. [00071] In Table 1, "Weighed amount of UC1010 (mg/g)" is the amount of 3- beta-hydroxy-5-alpha-pregnan-20-one per gram of final total composition including sterol (when a sterol is present). "Vehicle" denotes the carrier being tested. The amount of sterol is stated as "mg/gU, that is, the weight of sterol per weight of final total composition, including 3-beta-hydroxy-5-alpha-pregnan-20-one. "Appearance at preparation" describes the change of appearance of the mixture during preparation; usually the preparation is initially an emulsion or a solution whereas it has an oily appearance after evaporation of the ethanol; "Unchanged" denotes a sample that was stable and where thus 3-beta-hydroxy-5-alpha-pregnan-20-one remained in solution, without visible signs of haziness, precipitation, sedimentation, phase separation into two or more liquid phases or change of colour. This is also indicated by an asterisk () in the table. Table 1 3 4 5 6 7 8 9 10 1 1 12 0629-6 ACAO9 0629-4 ACAO9 0629-3 ACAO9 0629-5 ACAO9 0629-8 ACAO9 0709-2 ACAO9 0702-2 ACAO9 0822-8 ACAO9 0702-1 ACAO9 0822-A ACAO9 0822-C 10 10 20 10 3 5 5 5 5 5 + cholesterol (5,5 ms/s) Peanut oil Peanut oil + cholesterol (1 0 m9/9) Peanut oil + cholesterol (20 m9/9) Olive oil + cholesterol (1 0 ms/s) Sesame oil + cholesterol (3 ms/s) Sesame oil Sesame oil + cholesterol (2,5 ms/s) Sesame oil + cholesterol (5 ms/s) Sesame oil + cholesterol (5 ms/s) Sesame oil + cholesterol (1 0 ms/s) Emulsion to suspension Emulsion to oil (precipitation after a few hours) Emulsion to oil (precipitation after a few hours) Emulsion to oil Emulsion to oil Emulsion to oil Emulsion to oil Emulsion to oil Emulsion to oil Emulsion to oil Precipitation in almost the entire sample volume Bottom sediment Bottom sediment Bottom sediment Unchanged * Precipitation Unchanged * Unchanged * Unchanged * Unchanged * after 5 months Slight sedimentation after 13 months Bottom sediment Precipitation of crystals after 2 months A few particles on the bottom Precipitation of crystals after 2 months Unchanged after 1 2 months * 14 15 16 17 18 19 20 21 22 23 ACAO9 0822-D ACAO9 0822-F ACAO9 07 1 6-7 ACAO9 1024-1 ACAO9 1024-2 ACAO9 1025-1 ACAO9 1024-3 ACAO9 1024-4 ACAO9 1025-2 ACAO9 1025-3 10 1 1 10 10 1 0 15 10 10 15 15 Sesame oil + cholesterol (1 0 ms/s) Sesame oil + cholesterol (20 ms/s) Sesame oil + p-sitosterol (1 0 ms/s) Akomed R MCT Akomed R MCT + cholesterol (1 0 m9/9) Akomed R MCT + cholesterol (1 5 m9/9) 50 % Akomed R MCT, 50 % Akoline MCM 50 % Akomed R MCT, 50 % Akoline MCM + cholesterol (1 0 ms/s) 50 % Akomed R MCT, 50 % Akoline MCM 50 % Akomed R MCT, 50 % Akoline MCM + cholesterol (1 5 ms/s) Emulsion to oil Emulsion to oil Emulsion to oil Solution to oil Solution to oil Solution to oil Solution to oil Solution to oil Solution to oil Solution to oil Precipitation of crystals Precipitation of crystals Unchanged * Precipitation of crystals within an hour Slight precipitation of crystals on the bottom after 2 days Slight precipitation of crystals on the bottom after 24 hours Unchanged * Unchanged * Slight precipitation of crystals on the bottom after 1 week Unchanged * Crystals on the bottom an on the walls of the vial Unchanged after 10 months * Unchanged after 10 months * Unchanged after 10 months * MCT, 50 % Akoline 27 28 29 30 31 32 ACAO9 1 102-1 ACAO9 1025-4 ACAO9 1026-3 ACAO9 1101-2 ACAO9 1 109-1 ACAO9 1 109-2 30 15 20 20 25 30 + cholesterol (25 ms/s) 50 % Akomed R MCT, 50 % Akoline MCM + cholesterol (30 ms/s) Castor oil Castor oil Castor oil + cholesterol (20 ms/s) Castor oil + cholesterol (25 m9/9) Castor oil + cholesterol (3 1 m9/9) Solution to oil Solution to oil Solution to oil Solution to oil Solution to oil Solution to oil First sign of precipitation after 2 days Slight precipitation of crystals on the bottom after 26 days Slight precipitation of crystals on the bottom after 3 days Unchanged * Unchanged * Slight precipitation of crystals on the bottom after 8 days Unchanged after 10 months * Unchanged after 1 month, very slight precipitation after 2 months Unchanged after 1 week refrigerated * Small crystals on the bottom after 1 week refrigerated - unchanged after 2 days * 34 35 36 37 38 39 40 ACAO9 1 122-2 ACAO9 1101-4 ACAO9 1 102-2 ACAO9 1 108-1 ACAO9 1 108-2 ACAO9 1 120-3 ACAO9 1 120-1 40 25 30 34 38 40 40 Akoline MCM + cholesterol (40 ms/s) 50 % Castor oil , 50 % Akoline MCM + cholesterol (25 m9/9) 50 % Castor oil , 50 % Akoline MCM + cholesterol (30 m9/9) 48 % Castor oil , 52 % Akoline MCM + cholesterol (34 ms/s) 50 % Castor oil , 50 % Akoline MCM + cholesterol (38 m9/9) 50 % Castor oil , 50 % Akoline MCM + cholesterol (20 ms/s) 50 % Castor oil , 50 % Akoline MCM + cholesterol (40 ms/s) Solution to oil Solution to oil Solution to oil Solution to oil Solution to oil Solution to oil Solution to oil Unchanged after 24 hours * Unchanged * Unchanged * Unchanged after 15 days * Slight precipitation of crystals on the bottom after 9 days Massive precipitation after 24 hours Massive precipitation after 24 hours Precipitation after 1 week Unchanged after 10 months * Unchanged after 1 month, very slight precipitation after approx 2 months Precipitation in entire sample after 1 month After 1 week refrigerated, the entire sample was solidified, Slight precipitation after melting After 1 week refrigerated, the entire sample was solidified, Slight precipitation after melting Unchanged after 1 week refrigerated Small crystals on the bottom after 1 week refrigerated - unchanged after 2 days 46 47 48 49 ACAO9 1 2 16-2 ACAO9 1216-1 ACAO9 1221-2 ACAO9 1221-1 5 5 7,5 7,5 + cholesterol (75 m9/9) Olive oil Olive oil + cholesterol (1 0 ms/s) Olive oil Olive oil + cholesterol (7.5 m9/9) Emulsion to oil Emulsion to oil Emulsion to oil Emulsion to oil crystal sin the entire sample after 24 h Unchanged after 4 days, first signs of precipitation after 5 days Unchanged after 3 weeks * Precipitation after approx. 1 hour Unchanged after 1 day, very slight precipitation after 1 week Crystals at the bottom after 9 months Unchanged after 8 months Crystals at the bottom after 8 months Examples 50-75 [00072] Examples 50 to 75 were carried out essentially as examples 1 -49. Akoline Medium-Chain Monoglyceride (MCM) (Batch 8 192270 and 82 1 8940) and Akomed R Medium-Chain Triglyceride (MCT) (Batch 4765) were obtained from AarhusKarlshamns Sweden AB, Karlshamn, Sweden. Absolute ethanol (>99%) was obtained from VWR International. [00073] The procedure for making and evaluating lipid-based formulations was as follows: The batch sizes were either 20 g or 100 g of final formulation. The desired amounts of 3-beta-hydroxy-5-alpha-pregnan-20-onea nd cholesterol were weighed in a round-bottomed flask, 250 or 1000 ml depending on the batch size. [00074] To every gram of 3-beta-hydroxy-5-alpha-pregnan-20-onea nd cholesterol mixture a volume of about 15 to 30 ml of absolute ethanol was added. The smaller volume of alcohol per gram of solute was used when preparing the largest batch size of 1 00 g of final formulation. The mixture was treated in an ultrasonication bath (not exceeding 55°C) until a clear solution was obtained. This was normally accomplished within a few minutes. The glycerides were then added and the resulting mixture was treated in the ultrasonication bath for a few seconds until a clear, homogeneous liquid was obtained. The alcohol was evaporated from the liquid on a rotary evaporator at a pressure of about 20 mbar and a temperature of about 40°C until weight of the flask was more or less constant. Normally, the remaining ethanol content was 0.5% (w/w) or less. The evaporation time was 0.5-1.5 h, depending on the batch size. The aim was to obtain a practically uncolored liquid with the appearance of a clear oil at room temperature. The liquid was transferred to clear glass vials, which were stored at room temperature until evaluation. Some selected formulations were portioned and stored at 2-8°C for limited period of time. [00075] The evaluation comprised observation of the physical stability at room temperature over time. The samples were observed for haziness, precipitation of particles, aggregates or crystals, and subsequent sedimentation and/or phase separation into two or more liquid phases, and/or change of colour. [00076] It was possible to dissolve up to 25 mg of 3-beta-hydroxy-5-alpha-pregnan- 20-one per gram of final formulation based on 50% MCT and 50% MCM (examples 68 and 69) without any noticeable change in appearance when stored at room temperature for more than 1 month. One of the samples (example 69) also withstood storage at 2-8°C and repeated temperature cycling. [00077] In order to check the robustness and reproducibility of the formulations and the procedure for the preparation thereof, a scaling up of the batch size from 20 g to 100 g of final formulation was performed. The compositions corresponding to examples 60 and 69 were selected for this procedure. From the behavior during preparation and the initial observations of the resulting formulations (examples 74 and 75) it can be concluded that the adopted procedure is both robust and reproducible for making up to 100 g of formulation. Table 2 51 ACAlO 0 1 29-2 30 + cholesterol (30 mdgl 50 % Akoline MCM, 50 % Akomed R MCT + cholesterol (45 mdgl Solution to oil Signs of precipitation after 1 day Precipitation precipitation after 2 days --- --- -- 53 ACAlO 30 67 % Akoline Solution to oil Signs of slight Precipitation 0130-1 MCM, 33 % precipitation after 2 days Akomed R MCT + cholesterol (30 mg/g) 54 ACAlO 30 67 % Akoline Solution to oil Unchanged Precipitation 0 1 30-2 MCM, 33 % Akomed R MCT + cholesterol (45 mdgl 55 ACAlO 30 67 % Akoline Solution to oil Unchanged Precipitation 0 1 30-3 MCM, 33 % Akomed R MCT + cholesterol (60 mdgl 56 ACAlO 20 50 % Akoline Solution to oil Unchanged Unchanged after 29 Withstand 3 0203-8 MCM, 50 % weeks days in the Akomed R refrigerator, MCT but not 4 days, due to + cholesterol solidification of (40 mg/g) MCM 57 ACAlO 20 50 % Akoline Solution to oil Unchanged Unchanged after 29 Withstand 3 0203-C MCM, 50 % weeks weeks in the Akomed R refrigerator MCT + cholesterol (60 mdgl 58 ACAlO 20 50 % Akoline Solution to oil First signs of Precipitation 0204-1 MCM, 50 % precipitation after a few Akomed R hours, large star-shaped MCT crystals after 2 days + cholesterol (1 00 mg/gl 60 61 62 63 64 ACAl0 0206-1 ACAlO 0206-2 ACAlO 0206-3 ACAlO 0206-4 ACAlO 0206-5 20 20 20 20 20 (40 mdgl 30 % Akoline MCM, 70 % Akomed R MCT + cholesterol (60 mg/g) 30 % Akoline MCM, 70 % Akomed R MCT + cholesterol (1 00 mg/gl 15 % Akoline MCM, 85 % Akomed R MCT + cholesterol (40 mdgl 15 % Akoline MCM, 85 % Akomed R MCT + cholesterol (60 mdgl 15 % Akoline MCM, 85 % Akomed R MCT + cholesterol (1 00 mg/gl Solution to oil Solution to oil Solution to oil Solution to oil Solution to oil Unchanged First signs of precipitation after about 30 min Precipitation Unchanged after 1 days Signs of precipitation directly after evaporation Unchanged after 7 weeks Precipitation Precipitation Precipitation Precipitation Withstand 6 days in the refrigerator, but had precipitated after about 2 weeks without concomitant solidification of MCM Massive precipitation after 28 weeks 66 67 68 ACAlO 02 1 3-C ACAlO 02 1 3-8 ACAlO 0301-1 20 20 25 (40 mdgl 100 % Akomed R MCT + cholesterol (60 mdgl 100 % Akomed R MCT + cholesterol (80 mdgl 50 % Akoline MCM, 50 % Akomed R MCT + cholesterol (50 mdg) Solution to oil Solution to oil Solution to oil Precipitation in the whole sample Precipitation directly after evaporation Unchanged Precipitation in the whole sample Precipitation in the whole sample Unchanged A portion of the sample was placed in the refrigerator: partial solidification after 1 day, but clear solution after melting at RT. After another 1 day in the fridge the sample started to precipitate Massive precipitation after 25 weeks 70 71 72 ACAlO 030 1-3 ACAlO 030 1-4 ACAlO 030 1-5 25 30 30 + cholesterol (75 mdg) 50 % Akoline MCM, 50 % Akomed R MCT + cholesterol (1 00 mg/g) 50 % Akoline MCM, 50 % Akomed R MCT + cholesterol (60 mdg) 50 % Akoline MCM, 50 % Akomed R MCT + cholesterol (90 mdg) Solution to oil Solution to oil Solution to oil Very small amount of needle-shaped crystals after 1 day Unchanged after 3 days, slight tendency to precipitation after 5 days Unchanged after 3 days, precipitation after 5 days Precipitation Precipitation Precipitation partial solidification, but clear solution after melting at RT. The sample withstood repeated cycles during several weeks without precipitation! A portion of the sample was placed in the refrigerator: partial solidification and very slight precipitation after melting at RT A portion of the sample was placed in the refrigerator: partial solidification and slight precipitation after melting at RT A portion of the sample was placed in the refrigerator: partial solidification and substantial precipitation after melting at RT Example 76 74 75 [00078] The objective of the study was to investigate the comparative pharmacokinetics in plasma of a 3-beta-hydroxy-5-alpha-pregnan-20-onef ormulation comprising sesame oil and cholesterol following subcutaneous administration to New Zealand White rabbits. Two groups of three female rabbits each received single doses 1 mg/kg (formulation as in example 7) or 5 mg/kg (formulation as in example 10). Following subcutaneous injection into the dorsal neck region of the animals, blood samples were taken at 0.25, 0.5, 1, 2, 4, 6, 8, 1 2 and 24 hours post dose. Concentrations of 3-beta-hydroxy-5-alpha-pregnan-20-one in plasma were measured by a validated LC-MS/MS method. Data are presented in Figure 1 , and show mean plasma concentration (ng/mL) of 3-beta-hydroxy-5-alpha-pregnan-20-onefo r the two doses: 1 mg/kg (squares) and 5 mg/kg (circles). ACAlO 0320 ACAlO 032 1 25 20 (1 20 mg/g) 50 % Akoline MCM, 50 % Akomed R MCT + cholesterol (75 mg/g) 30 % Akoline MCM, 70 % Akomed R MCT + cholesterol (60 mg/g) discarded) Solution to oil Solution to oil Unchanged Unchanged Unchanged Unchanged Batch size 100 g Very slight precipitation after 23 weeks Batch size 100 g Slight precipitation after 23 weeks Example 77 [00079] Suspensions of 3-beta-hydroxy-5-alpha-pregnan-20-onew ere prepared for investigation of solubility as follows: cholesterol was first dissolved at room temperature in sesame oil at 10 and 20 mg/ml, respectively. Suspensions of 3-beta-hydroxy-5- alpha-pregnan-20-one in sesame oil with different amounts of cholesterol were prepared on a magnetic stirrer at about 500 rpm using a conventional Teflon coated stir bar, at room temperature for several days, with occasional cycling to 2-8°C. At this time, the particles have become substantially smaller. Thereafter, samples of the respective suspensions were filtered through a 0.2 pm filter and analyzed with regard to concentration of 3-beta-hydroxy-5-alpha-pregnan-20-one.T he results are presented in Table 3 and Figure 2 where it can be seen that increasing the amounts of cholesterol increases the soluble fraction of 3-beta-hydroxy-5-alpha-pregnan-20-one. Figure 2 shows data for a 3-beta-hydroxy-5-alpha-pregnan-20-onec oncentration of 1 0 mg/g. Figure 3a and 3 b show microscopy pictures taken at 5x enlargement of suspensions with a 3-beta-hydroxy-5-alpha-pregnan-20-onec oncentration of 1 0 mg/g and a cholesterol:3-beta-hydroxy-5-alpha-pregnan-20-one ratio of 1 : 1 immediately upon mixing (3a) and after several days of stirring (3b). Table 3 Example 78 [00080] Micronized 3-beta-hydroxy-5-alpha-pregnan-20-one (1 0 mg/g) with a mean particle size of 6 micrometer was suspended in sesame oil with cholesterol (20 mg/g) and stirred as in example 77. Photos were taken immediately upon suspension (Fig 4a) and after 19 hours of stirring (Fig 4b). Conclusion [0008 11 It has been shown that the presence of a sterol such as cholesterol improves the possibilities to formulate 3-beta-hydroxy-5-alpha-pregnan-20-one in a pharmaceutical composition comprising acylglycerols, either as an oily solution or an oily suspension. CLAIMS 1. A pharmaceutical composition comprising 3-beta-hydroxy-5-alpha-pregnan- 20-one, at least one sterol or an ester thereof and a mixture of acylglycerols with a solid fat content of less than about 25% at 25°C and about 0% at 37°C. 2. The pharmaceutical composition according to clam 1 wherein the sterol is selected from the group consisting of cholesterol, beta-sitosterol, stigmasterol, brassicasterol and avenasterol. 3. The pharmaceutical composition according to claim 2 wherein the sterol is cholesterol. 4. The pharmaceutical composition according to any one of claims 1 to 3 wherein the mixture of acylglycerols is a vegetable oil. 5. The pharmaceutical composition according to claim 4 where the vegetable oil is selected from the group consisting of sesame oil, peanut oil, olive oil, and castor oil. 6. The pharmaceutical composition according to any one of claims 1 to 3 wherein the mixture of acylglycerols is a medium-chain acylglycerol. 7. The pharmaceutical composition according to any one of claims 1 to 6 in which 3-beta-hydroxy-5-alpha-pregnan-20-one is essentially dissolved. 8. The pharmaceutical composition according to any one of claims 1 to 6 comprising a suspension of 3-beta-hydroxy-5-alpha-pregnan-20-one. 9. A pharmaceutical composition for parenteral administration according to any one of claims 1 to 8. 10. A pharmaceutical composition for oral administration according to any one of claims 1 to 8. 1 1 . A pharmaceutical composition for vaginal administration according to any one of claims 1 to 8. 12. A pharmaceutical composition for nasal administration according to any one of claims 1 to 8. 13. A method for preparing a pharmaceutical composition according to claim 7 comprising the steps of a) dissolving 3-beta-hydroxy-5-alpha-pregnan-20-one in ethanol, b) adding a mixture of acylglycerols with a solid fat content of less than about 25% at 25°C and about 0% at 37°C and a sterol or an ester thereof, c) mixing until a homogeneous liquid is obtained and d) evaporating the ethanol. 14. A pharmaceutical composition obtainable by the method according to claim 13. 15. Use of a pharmaceutical composition according to any one of claims 1 to 12 or claim 14 for the treatment or prevention of conditions of the central nervous system. 16. The use according to claim 15 where the condition of the central nervous system is selected from the group consisting of epilepsy, menstruation cycle dependant epilepsy, depression, stress related depression, migraine, tiredness and in particular stress related tiredness, premenstrual syndrome, premenstrual dysphoric disorder, menstrual cycle linked mood changes, stress related memory changes, menstrual cycle linked memory changes, Alzheimer's dementia, menstrual cycle linked difficulties in concentration, menstrual cycle linked sleep disorders and tiredness, substance abuse, menstrual cycle linked alcoholism, side effects of oral contraceptives and postmenopausal therapy or combinations thereof 17. Use of the pharmaceutical composition according to any one of claims 1 to 12 or claim 14 for the termination of steroid induced anaesthesia.

Full Text

A pharmaceutical composition comprising 3-beta-hydroxy-5-alpha-pregnan-20-one
with improved storage and solubility properties
Technical field
[0001] The present invention relates generally to an improved formulation of 3-
beta-hydroxy-5-alpha-pregnan-20-one.
Background art
[0002] 3-beta-hydroxy-5-alpha-pregnan-20-one is a steroid in the pregnane
family and a modulator of GABA,-receptor activity which is indicated for the treatment
of sex/stress steroid induced disorders conditions (W099/4593 1 ). 3-beta-hydroxy-5-
alpha-pregnan-20-one is poorly soluble in many therapeutically acceptable solvents,
which makes it difficult to administer the compound to a patient.
[0003] In animal studies, 3-beta-hydroxy-5-alpha-pregnan-20-one has been
intravenously administered to rats in a formulation containing cyclodextrin
(W099/4593 1 ).
[0004] Grant et al (JPET 326:354-362, 2008) has administered 3-beta-hydroxy-5-
alpha-pregnan-20-one to monkeys by using a formulation with hydroxypropyl Pcyclodextrin.
[0005] Formulations with cyclodextrins are not suitable for administration to
human patients. One reason for this is, because 3-beta-hydroxy-5-aIpha-pregnan-20-
one is poorly soluble, the formulation results in a large therapeutic volume that can
only be administered intravenously.
[0006] Since 3-beta-hydroxy-5-alpha-pregnan-20-one is poorly soluble in water
there is still no pharmaceutically acceptable formulation for this compound.
Definitions
[0007] As used in the present application, the following terms have the specified
meanings unless otherwise specified.
[0008] By "acylglycerol" is meant all types and combinations of fatty acids
esterified to glycerol.
[0009] By "medium-chain acylglycerol" is meant a mixture of acylglycerols where
the total com bined percentage of octanoic (caprylic) acid and decanoic (capric) acid
is at least 95%.
[00010] By "solid fat content" is meant the percentage of solid as determined by
pulse NMR (nuclear magnetic resonance).
[0001 11 "Room temperature" denotes a temperature of between 1 8°C and 25°C.
[000 1 21 "UC 1 0 1 0"denotes 3-beta-hydroxy-5-alpha-pregnan-20-one.
[000 1 31 "Sterol or ester thereof" denotes steroids with at least one hydroxyl group
and esters of said steroids where at least one hydroxyl group has been used for the
synthesis of an ester.
[00014] Steroids, such as sterols, are usually described by the number of carbon
atoms in the compound. Thus, for example, cholesterol is a C27 sterol, which indicates
that the compound consists of 27 carbon atoms.
[000 1 51 Unless stated otherwise, concentrations are stated as mg/g, that is, mg
per gram of pharmaceutical composition.
Brief description of fig- ures
[000 1 61 Figure 1 shows mean plasma concentrations of 3-beta-hydroxy-5-alphapregnan-
20-one (ng/mL) in rabbits following subcutaneous administration of 3-betahydroxy-
5-alpha-pregnan-20-one in sesame oil with cholesterol (1 : 1 ) in two doses: 1
mg/kg (squares) and 5 mg/kg (circles).
[000 1 7] Figure 2 shows 3-beta-hydroxy-5-alpha-pregnan-20-onec oncentration in
filtrate (0.2 pm) from a suspension of 3-beta-hydroxy-5-alpha-pregnan-20-one in
relation to cholesterol:3-beta-hydroxy-5-aIpha-pregnan-20-one ratio.
[00018] Figure 3a, 3 b, 4a and 4b show photographs of suspensions of 3-betahydroxy-
5-alpha-pregnan-20-one.
Summary of invention
[00019] An object of the present invention is to provide an improved formulation of
3-beta-hydroxy-5-alpha-pregnan-20-one in a pharmaceutically acceptable carrier.
[00020] Another object of the present invention is to provide a formulation of 3-
beta-hydroxy-5-alpha-pregnan-20-onew ith enhanced storage properties.
[00021] Yet another object is to provide a formulation of 3-beta-hydroxy-5-alphapregnan-
20-one with improved pharmacokinetics.
[00022] Yet another object of the present invention is to provide a formulation of 3-
beta-hydroxy-5-alpha-pregnan-20-onew ith increased solubility in a pharmacologically
acceptable carrier.
[00023] These and other objects are met by a first general aspect of the present
invention which provides a pharmaceutical formulation comprising 3-beta-hydroxy-5-
alpha-pregnan-20-one, at least one sterol or an ester thereof and a mixture of
acylglycerols with a solid fat content of less than about 25% at 25°C and about 0% at
37°C.
[00024] In a second general aspect of the present invention there is provided
methods for preparing a pharmaceutical composition.
[00025] In a third general aspect of the present invention there is provided a
pharmaceutical composition obtainable according to a method according to the
invention.
[00026] In a fourth general aspect of the present invention there is provided use of
a pharmaceutical composition for the treatment of conditions of the central nervous
system.
Detailed description
[00027l The inventors have found that the addition of a sterol surprisingly
increases the solubility and improves the pharmacokinetics of 3-beta-hydroxy-5-alphapregnan-
20-one in acylglycerols.
[0002 81 Generally, the pharmaceutical composition comprises 3-beta-hydroxy-5-
alpha-pregnan-20-one, at least one sterol or an ester thereof and a mixture of
acylglycerols with a solid fat content of less than about 25% at 25°C and about 0% at
[00029] In particular a sterol with a hydroxyl group bound to the third carbon
atom of the sterol structure is useful in the invention. The sterol may be cholesterol or
beta-sitosterol, but also other sterols such as stigmasterol, brassicasterol or avenasterol
may be used. In particular, cholesterol may be used.
[00030] In addition, cholesteryl esters can be used. Examples of such esters are
sodium cholesteryl sulphate, cholesteryl bensoate, cholesteryl acetate, cholesteryl
caprylate, cholesteryl decanoate, cholestyl palmitate, cholesteryl oleate and cholesteryl
steara te.
[00031] The sterol or ester thereof can be a C18-C30 sterol or an ester thereof, a
C21 -C27 sterol or an ester thereof, or a C27-C29 sterol or ester thereof.
[00032] The pharmaceutical composition can, in a first embodiment, be such that
3-beta-hydroxy-5-alpha-pregnan-20-one is essentially dissolved in the composition. Thus
3-beta-hydroxy-5-alpha-pregnan-20-onec an be dissolved or essentially dissolved
according to this embodiment of the invention.
[00033] The weight ratio of sterol (or ester thereof) to 3-beta-hydroxy-5-alphapregnan-
20-one can, in this embodiment, be in the range of about 1 : 10 to 10: 1 . The
sterol or ester thereof may be added in an amount that is similar to the amount of 3-
beta-hydroxy-5-alpha-pregnan-20-one by weight. Because 3-beta-hydroxy-5-alphapregnan-
20-one and a sterol have similar molecular weights this results in almost
equimolar amounts of 3-beta-hydroxy-5-alpha-pregnan-20-oneto sterol.
[00034] Thus, the weight ratio of sterol to 3-beta-hydroxy-5-alpha-pregnan-20-one
can be in the range of from 1 :5 to 5: 1 . In particular, the weight ratio of sterol to 3-
beta-hydroxy-5-alpha pregnan-20-one can be from 1 :3 to 3: 1 .
[00035] Suitable concentrations of 3-beta-hydroxy-5-alpha pregnan-20-one are
between 0.1 mg/g and 75 mg/g. The concentration of 3-beta-hydroxy-5-alpha
pregnan-20-one can also be between 1 mg/g and 50 mg/g, between 5 mg/g and
30 mg/g or between 10 mg/g and 25 mg/g.
[00036] Alternatively, in a second embodiment, the pharmaceutical composition
comprises a suspension of 3-beta-hydroxy-5-alpha-pregnan-20-one. In this case the
pharmaceutical composition will comprise 3-beta-hydroxy-5-alpha-pregnan-20-one in
particles as well as 3-beta-hydroxy-5-alpha-pregnan-20-oned issolved in the
composition. The sterol increases the soluble fraction of 3-beta-hydroxy-5-alphapregnan-
20-one in such a suspension compared to a suspension without a sterol. One
advantage with a suspension is that the formulation can contain a high concentration
of 3-beta-hydroxy-5-alpha-pregnan-20-one.A n additional advantage with a
composition that comprises a suspension is that it results in slow release of 3-betahydroxy-
5-alpha-pregnan-20-one.
[00037] When the pharmaceutical composition comprises a suspension, the
particles are preferably of a range of sizes that is not engulfed by macrophages.
Macrophages do primarily engulf particles of a size that is 2-3 micrometer (Champion
et al, Pharm Res 2008; 25(8): 1 8 15-1 82 1 ).
[00038] In this second embodiment, the weight ratio of sterol (or ester thereof) to 3-
beta-hydroxy-5-alpha pregnan-20-one can be in the range of about 1 : 1 0 to 1 0: 1 . The
weight ratio of sterol to 3-beta-hydroxy-5aIpha pregnan-20-one can be in the range of
from 1 :5 to 5: 1 . In particular, the weight ratio of sterol to 3-beta-hydroxy-5-alphapregnan-
20-one can be from 1 :4 to 3:1 or from 1 :3 to 3:1.
[00039] Suitable concentrations of 3-beta-hydroxy-5-alpha-pregnan-20-onea re, in
this second embodiment, between 0.1 mg/g and 750 mg/g. The concentration of 3-
beta-hydroxy-5-alpha-pregnan-20-one can also be between 1 mg/g and 300 mg/g,
between 1 mg/g and 100 mg/g, between 1 mg/g and 50 mg/g, between 5 mg/g
and 30 mg/g or between 10 mg/g and 25 mg/g.
[00040] The following applies to the invention in general.
[0004 11 Generally, the mixture of acylglycerols is characterized in that it has a
solid fat content of less than about 25% at 25°C and about 0% at 37°C. Thus, the
solid fat content is, for practical purposes, 0% at 37°C. The solid fat content is at most
0.01 % at 37°C.
[00042] The mixture of acylglycerols can be a vegetable oil. Thus, it can be a
vegetable oil selected from the group consisting of sesame oil, peanut oil, olive oil,
and castor oil, or mixtures thereof.
[00043] In particular the mixture of acylglycerols can be a medium-chain
acylglycerol, that is, a mixture of acylglycerols wherein the total combined percentage
of fatty acids with 8 carbon atoms (octanoic acid) and 10 carbon atoms (decanoic
acid) is at least 95%. The medium-chain acylglycerol can be various mixtures of
monoacylglycerols, diacylglycerols and triacylglycerols.
[00044] The medium-chain acylglycerol can consist of from about 50% to about
65% of monoacylglycerols, about 25% to about 35% of diacylglycerols, less than
about 5% of triacylglycerols and less than about 2.5% of glycerol. An example of such
a medium chain acylglycerol is Akoline MCM.
[00045] The medium-chain acylglycerol can be such that it comprises at least about
95% triacylglycerols. Akomed R MCT is an example of such a medium-chain
acylglycerol.
[00046] The mixture of acylglycerols can comprise a mixture of a vegetable oil and
a medium-chain acylglycerol. The mixture of acylglycerols can comprises a mixture of
castor oil and a medium-chain acylglycerol where castor oil is present in an amount of
between 40% and 60% by weight. The mixture of acylglycerols can consist of about
48% by weight of castor oil and about 52% by weight of a medium-chain
acylglycerol. In particular the mixture of acylglycerols can consist of about 48% by
weight castor oil and about 52% by weight of a medium-chain acylglycerol.
[00047] The pharmaceutical composition may comprise additional excipients
known to a person skilled in the art such as antioxidants, preservatives, surfactants,
coloring, flavoring, or thickening agents.
[00048] The pharmaceutical composition can be administered to the patient by
different means. Thus, it may be administered orally, parenterally or topically. Thus, the
pharmacological composition may be administered subcutaneously, intramuscularly,
intravenously, nasally, transdermally or vaginally.
[00049] In a second general aspect of the present invention there is provided
methods for preparing a pharmaceutical composition of 3-beta-hydroxy-5-alphapregnan-
20-one.
[00050] One method, in which 3-beta-hydroxy-5-alpha-pregnan-20-one is dissolved
or essentially dissolved in the composition, comprises the steps of a) dissolving 3-betahydroxy-
5-alpha-pregnan-20-one in ethanol, b) adding a mixture of acylglycerols with
a solid fat content of less than about 25% at 25°C and about 0% at 37°C and a sterol
or ester thereof, c) mixing until a homogeneous liquid is obtained and d) evaporating
the ethanol.
[00051] When the mixture of acylglycerol is a solid or a semi-solid at room
temperature, such as a medium-chain acylglycerol, the method may comprise a further
step, which is the melting of the medium-chain acylglycerol before mixing it with the
ethanol-drug preparation. The melting step enables the homogeneous mixing of this
type of acylglycerol with other components. Once melted and mixed with the other
components, the preparation remains in a liquid state for at least the time periods
indicated in Table 1.
[00052] When the formulation comprises a suspension, the formulation is
advantageously prepared according to a method that comprises the following steps: 1 )
dissolving or suspending the sterol or ester thereof in the mixture of acylglycerols, 2)
suspending 3-beta-hydroxy-5-alpha-pregnan-20-one in the acylglycerol-sterol mixture,
3) gently mixing. Surprisingly, this procedure leads to suspended particles comprising
3-beta-hydroxy-5-alpha-pregnan-20-oneo f smaller size.
[00053] In a third general aspect of the present invention there is provided
pharmaceutical compositions obtainable by the methods according to the second
aspect of the invention.
[00054] In a fourth general aspect of the present invention there is provided the
use of the pharmaceutical composition according to the invention for the treatment or
prevention of conditions of the central nervous system.
[00055] The pharmaceutical composition can be used to treat or prevent conditions
of the central nervous system. Examples of such conditions that can be treated are
epilepsy, menstruation cycle dependant epilepsy, depression, stress related
depression, migraine, tiredness and in particular stress related tiredness, premenstrual
syndrome, premenstrual dysphoric disorder, menstrual cycle linked mood changes,
stress related memory changes, menstrual cycle linked memory changes, Alzheimer's
dementia, menstrual cycle linked difficulties in concentration, menstrual cycle linked
sleep disorders and tiredness, substance abuse, menstrual cycle linked alcoholism, or
com binations thereof.
[00056] In particular the pharmaceutical composition can be used to treat steroid
induced mood disorders, in particular premenstrual dysphoric disorder.
[00057] The pharmaceutical composition can also be used for treating or
preventing side effects of oral contraceptives and postmenopausal therapy.
[00058] The pharmaceutical composition can also be used for control or
termination of steroid-induced anesthesia.
Examples
[00059] The invention will now be described with non-limiting examples.
[00060] In order to find a pharmaceutical composition comprising 3-beta-hydroxy-
5-alpha-pregnan-20-one, several different vehicles and com binations of vehicles were
evaluated. 3-beta-hydroxy-5-alpha-pregnan-20-onew as dissolved in various vehicles
and was evaluated by eye for physical stability at room temperature over time. A
formulation that did not undergo any visible changes in appearance and remained
clear on storage in room temperature without signs of haziness, precipitation,
sedimentation, phase separation into two or more liquid phases or change of colour,
within 30 days was considered as "stable".
[00061] In addition, the particle size and solubility of 3-beta-hydroxy-5-alphapregnan-
20-one in formulations comprising a suspension of 3-beta-hydroxy-5-alphapregnan-
20-one were evaluated.
General procedure for examples 1-49
[00062] The following procedure was adopted for the preparation of 3-betahydroxy-
5-alpha-pregnan-20-one-containing formulations.
[00063] The desired amount of 3-beta-hydroxy-5-alpha-pregnan-20-onea nd sterol
(for example cholesterol) was weighed in a 100 ml or 250 ml round-bottomed flask.
To every gram of the mixture of 3-beta-hydroxy-5-alpha-pregnan-20-one and sterol a
volume of about 30 ml of absolute ethanol was added. The mixture was treated in an
ultrasonication bath (not exceeding 50°C) until a clear liquid was obtained. This was
normally accomplished within 10 minutes. The additional lipid ingredients indicated in
the "Vehicle" column in Table 1 was then added up to 20 g. The resulting mixture was
gently shaken by hand until a clear, homogeneous liquid was obtained. When the
lipid was a solid at room temperature it was melted in the warm ultrasonication bath to
a liquid form before addition.
[00064] Compounds from the following suppliers were used (product numbers
within brackets): Cholesterol from Sigma (C8503), olive oil and peanut oil from
Apoteket, Sweden (26 36 16 and 26 66 01, respectively), sesame oil from Fluka
(85067), and castor oil from Sigma (259853). Akomed R medium-chain triacylglycerol
(MCT) and Akoline medium-chain monoacylglycerol (MCM) were both from
AarhusKarlshamns Sweden AB, Karlshamn, Sweden.
[00065] The alcohol was evaporated from the liquid on a rotary evaporator at a
pressure of about 25 mbar and a temperature of about 40°C until the weight of the
flask was essentially constant. The remaining ethanol content was essentially less than
1 %. The aim was to obtain a liquid with the appearance of clear oil at room
temperature. The oily liquid was then transferred to clear glass vials and stored at
room temperature until evaluation.
[00066] The samples were evaluated by observing the samples in the glass vials
and recording signs of haziness, precipitation, sedimentation, phase separation into
two or more liquid phases, or change of colour after 1 or 2 days after preparation and
after 30 days after preparation. In some cases, other time intervals were used
(indicated in Table 1). Where indicated, the entire sample was placed in a refrigerator
(2-8°C) to provoke precipitation.
Example 1
[00067] 3-beta-hydroxy-5-alpha-pregnan-20-one (UC 1 0 1 0) (5 mg/g) and peanut
oil was mixed into an emulsion with ethanol as described above in the concentrations
shown in Table 1, and the ethanol was evaporated. The final weight of the
preparation was 20 g. The mixture then had the form of an oily liquid. After one day
when the sample was evaluated there were signs of precipitation. At 30 days the
precipitate has formed a bottom sediment. Thus, the formulation was not stable.
Example 2
[00068] Example 2 was carried out essentially as Example 1 with the difference
that cholesterol (5.5 mg/g) was added. When the sample was evaluated after one
day the appearance of the sample had not changed. It was still unchanged after 30
days and after four months. After five months there was a slight precipitation. Example
2 compared with Example 1 shows how the addition of 5.5 mg/g cholesterol to a
solution of 5 mg/g 3-beta-hydroxy-5-alpha-pregnan-20-one in peanut oil substantially
increases the solubility so that, instead of precipitating, no precipitation occurred and
the sample was stable for four months. However, a slight sedimentation occurred after
5 months.
Examples 3 to 49
[00069] Examples 3 to 49 were carried out essentially as described above with
the variations with regard to 3-beta-hydroxy-5-alpha-pregnan-20-onec oncentration,
acylglycerol mixture used, sterol used and sterol concentration that are shown in Table
1.
[00070] The data from examples 1 to 49 is presented in Table 1 . The effect of
addition of cholesterol is, for example, evident in examples 8 and 12, where the
addition of cholesterol (1 0 mg/g) substantially increased solubility so that the sample
did not precipitate but instead was stable for 12 months.
[00071] In Table 1, "Weighed amount of UC1010 (mg/g)" is the amount of 3-
beta-hydroxy-5-alpha-pregnan-20-one per gram of final total composition including
sterol (when a sterol is present). "Vehicle" denotes the carrier being tested. The amount
of sterol is stated as "mg/gU, that is, the weight of sterol per weight of final total
composition, including 3-beta-hydroxy-5-alpha-pregnan-20-one. "Appearance at
preparation" describes the change of appearance of the mixture during preparation;
usually the preparation is initially an emulsion or a solution whereas it has an oily
appearance after evaporation of the ethanol; "Unchanged" denotes a sample that was
stable and where thus 3-beta-hydroxy-5-alpha-pregnan-20-one remained in solution,
without visible signs of haziness, precipitation, sedimentation, phase separation into
two or more liquid phases or change of colour. This is also indicated by an asterisk (*)
in the table.
Table 1
3
4
5
6
7
8
9
10
1 1
12
0629-6
ACAO9
0629-4
ACAO9
0629-3
ACAO9
0629-5
ACAO9
0629-8
ACAO9
0709-2
ACAO9
0702-2
ACAO9
0822-8
ACAO9
0702-1
ACAO9
0822-A
ACAO9
0822-C
10
10
20
10
3
5
5
5
5
5
+ cholesterol (5,5
ms/s)
Peanut oil
Peanut oil
+ cholesterol (1 0
m9/9)
Peanut oil
+ cholesterol (20
m9/9)
Olive oil
+ cholesterol (1 0
ms/s)
Sesame oil
+ cholesterol (3
ms/s)
Sesame oil
Sesame oil
+ cholesterol (2,5
ms/s)
Sesame oil
+ cholesterol (5
ms/s)
Sesame oil
+ cholesterol (5
ms/s)
Sesame oil
+ cholesterol (1 0
ms/s)
Emulsion to
suspension
Emulsion to oil
(precipitation
after a few
hours)
Emulsion to oil
(precipitation
after a few
hours)
Emulsion to oil
Emulsion to oil
Emulsion to oil
Emulsion to oil
Emulsion to oil
Emulsion to oil
Emulsion to oil
*
Precipitation in
almost the entire
sample volume
Bottom sediment
Bottom sediment
Bottom sediment
Unchanged
*
Precipitation
Unchanged
*
Unchanged
*
Unchanged
*
Unchanged
*
after 5 months
Slight sedimentation
after 13 months
Bottom sediment
Precipitation of
crystals after 2
months
A few particles on the
bottom
Precipitation of
crystals after 2
months
Unchanged after 1 2
months
*
14
15
16
17
18
19
20
21
22
23
ACAO9
0822-D
ACAO9
0822-F
ACAO9
07 1 6-7
ACAO9
1024-1
ACAO9
1024-2
ACAO9
1025-1
ACAO9
1024-3
ACAO9
1024-4
ACAO9
1025-2
ACAO9
1025-3
10
1 1
10
10
1 0
15
10
10
15
15
Sesame oil
+ cholesterol (1 0
ms/s)
Sesame oil
+ cholesterol (20
ms/s)
Sesame oil
+ p-sitosterol (1 0
ms/s)
Akomed R MCT
Akomed R MCT
+ cholesterol (1 0
m9/9)
Akomed R MCT
+ cholesterol (1 5
m9/9)
50 % Akomed R
MCT, 50 % Akoline
MCM
50 % Akomed R
MCT, 50 % Akoline
MCM
+ cholesterol (1 0
ms/s)
50 % Akomed R
MCT, 50 % Akoline
MCM
50 % Akomed R
MCT, 50 % Akoline
MCM
+ cholesterol (1 5
ms/s)
Emulsion to oil
Emulsion to oil
Emulsion to oil
Solution to oil
Solution to oil
Solution to oil
Solution to oil
Solution to oil
Solution to oil
Solution to oil
Precipitation of
crystals
Precipitation of
crystals
Unchanged
*
Precipitation of
crystals within an
hour
Slight precipitation
of crystals on the
bottom after 2
days
Slight precipitation
of crystals on the
bottom after 24
hours
Unchanged
*
Unchanged
*
Slight precipitation
of crystals on the
bottom after 1
week
Unchanged
*
Crystals on the
bottom an on the
walls of the vial
Unchanged after 10
months
*
Unchanged after 10
months
*
Unchanged after 10
months
*
MCT, 50 % Akoline
27
28
29
30
31
32
ACAO9
1 102-1
ACAO9
1025-4
ACAO9
1026-3
ACAO9
1101-2
ACAO9
1 109-1
ACAO9
1 109-2
30
15
20
20
25
30
+ cholesterol (25
ms/s)
50 % Akomed R
MCT, 50 % Akoline
MCM
+ cholesterol (30
ms/s)
Castor oil
Castor oil
Castor oil
+ cholesterol (20
ms/s)
Castor oil
+ cholesterol (25
m9/9)
Castor oil
+ cholesterol (3 1
m9/9)
Solution to oil
Solution to oil
Solution to oil
Solution to oil
Solution to oil
Solution to oil
First sign of
precipitation after
2 days
Slight precipitation
of crystals on the
bottom after 26
days
Slight precipitation
of crystals on the
bottom after 3
days
Unchanged
*
Unchanged
*
Slight precipitation
of crystals on the
bottom after 8
days
Unchanged after 10
months
*
Unchanged after 1
month, very slight
precipitation after 2
months
Unchanged
after 1 week
refrigerated
*
Small crystals
on the
bottom after
1 week
refrigerated
- unchanged
after 2 days
*
34
35
36
37
38
39
40
ACAO9
1 122-2
ACAO9
1101-4
ACAO9
1 102-2
ACAO9
1 108-1
ACAO9
1 108-2
ACAO9
1 120-3
ACAO9
1 120-1
40
25
30
34
38
40
40
Akoline MCM
+ cholesterol (40
ms/s)
50 % Castor oil , 50
% Akoline MCM
+ cholesterol (25
m9/9)
50 % Castor oil , 50
% Akoline MCM
+ cholesterol (30
m9/9)
48 % Castor oil , 52
% Akoline MCM
+ cholesterol (34
ms/s)
50 % Castor oil , 50
% Akoline MCM
+ cholesterol (38
m9/9)
50 % Castor oil , 50
% Akoline MCM
+ cholesterol (20
ms/s)
50 % Castor oil , 50
% Akoline MCM
+ cholesterol (40
ms/s)
Solution to oil
Solution to oil
Solution to oil
Solution to oil
Solution to oil
Solution to oil
Solution to oil
Unchanged after
24 hours
*
Unchanged
*
Unchanged
*
Unchanged after
15 days
*
Slight precipitation
of crystals on the
bottom after 9
days
Massive
precipitation after
24 hours
Massive
precipitation after
24 hours
Precipitation after 1
week
Unchanged after 10
months
*
Unchanged after 1
month, very slight
precipitation after
approx 2 months
Precipitation in entire
sample after 1 month
After 1 week
refrigerated,
the entire
sample was
solidified,
Slight
precipitation
after melting
After 1 week
refrigerated,
the entire
sample was
solidified,
Slight
precipitation
after melting
Unchanged
after 1 week
refrigerated
Small crystals
on the
bottom after
1 week
refrigerated
- unchanged
after 2 days
46
47
48
49
ACAO9
1 2 16-2
ACAO9
1216-1
ACAO9
1221-2
ACAO9
1221-1
5
5
7,5
7,5
+ cholesterol (75
m9/9)
Olive oil
Olive oil
+ cholesterol (1 0
ms/s)
Olive oil
Olive oil
+ cholesterol (7.5
m9/9)
Emulsion to oil
Emulsion to oil
Emulsion to oil
Emulsion to oil
crystal sin the
entire sample after
24 h
Unchanged after
4 days, first signs
of precipitation
after 5 days
Unchanged after
3 weeks *
Precipitation after
approx. 1 hour
Unchanged after
1 day, very slight
precipitation after
1 week
Crystals at the bottom
after 9 months
Unchanged after 8
months
Crystals at the bottom
after 8 months
Examples 50-75
[00072] Examples 50 to 75 were carried out essentially as examples 1 -49.
Akoline Medium-Chain Monoglyceride (MCM) (Batch 8 192270 and 82 1 8940) and
Akomed R Medium-Chain Triglyceride (MCT) (Batch 4765) were obtained from
AarhusKarlshamns Sweden AB, Karlshamn, Sweden. Absolute ethanol (>99%) was
obtained from VWR International.
[00073] The procedure for making and evaluating lipid-based formulations was as
follows: The batch sizes were either 20 g or 100 g of final formulation. The desired
amounts of 3-beta-hydroxy-5-alpha-pregnan-20-onea nd cholesterol were weighed in a
round-bottomed flask, 250 or 1000 ml depending on the batch size.
[00074] To every gram of 3-beta-hydroxy-5-alpha-pregnan-20-onea nd cholesterol
mixture a volume of about 15 to 30 ml of absolute ethanol was added. The smaller
volume of alcohol per gram of solute was used when preparing the largest batch size
of 1 00 g of final formulation. The mixture was treated in an ultrasonication bath (not
exceeding 55°C) until a clear solution was obtained. This was normally accomplished
within a few minutes. The glycerides were then added and the resulting mixture was
treated in the ultrasonication bath for a few seconds until a clear, homogeneous liquid
was obtained. The alcohol was evaporated from the liquid on a rotary evaporator at a
pressure of about 20 mbar and a temperature of about 40°C until weight of the flask
was more or less constant. Normally, the remaining ethanol content was 0.5% (w/w)
or less. The evaporation time was 0.5-1.5 h, depending on the batch size. The aim
was to obtain a practically uncolored liquid with the appearance of a clear oil at room
temperature. The liquid was transferred to clear glass vials, which were stored at room
temperature until evaluation. Some selected formulations were portioned and stored at
2-8°C for limited period of time.
[00075] The evaluation comprised observation of the physical stability at room
temperature over time. The samples were observed for haziness, precipitation of
particles, aggregates or crystals, and subsequent sedimentation and/or phase
separation into two or more liquid phases, and/or change of colour.
[00076] It was possible to dissolve up to 25 mg of 3-beta-hydroxy-5-alpha-pregnan-
20-one per gram of final formulation based on 50% MCT and 50% MCM (examples
68 and 69) without any noticeable change in appearance when stored at room
temperature for more than 1 month. One of the samples (example 69) also withstood
storage at 2-8°C and repeated temperature cycling.
[00077] In order to check the robustness and reproducibility of the formulations
and the procedure for the preparation thereof, a scaling up of the batch size from 20
g to 100 g of final formulation was performed. The compositions corresponding to
examples 60 and 69 were selected for this procedure. From the behavior during
preparation and the initial observations of the resulting formulations (examples 74 and
75) it can be concluded that the adopted procedure is both robust and reproducible
for making up to 100 g of formulation.
Table 2
51 ACAlO
0 1 29-2
30
+ cholesterol
(30 mdgl
50 % Akoline
MCM, 50 %
Akomed R
MCT
+ cholesterol
(45 mdgl
Solution to oil Signs of precipitation
after 1 day
Precipitation
precipitation
after 2 days
--- --- --
53 ACAlO 30 67 % Akoline Solution to oil Signs of slight Precipitation
0130-1 MCM, 33 % precipitation after 2 days
Akomed R
MCT
+ cholesterol
(30 mg/g)
54 ACAlO 30 67 % Akoline Solution to oil Unchanged Precipitation
0 1 30-2 MCM, 33 %
Akomed R
MCT
+ cholesterol
(45 mdgl
55 ACAlO 30 67 % Akoline Solution to oil Unchanged Precipitation
0 1 30-3 MCM, 33 %
Akomed R
MCT
+ cholesterol
(60 mdgl
56 ACAlO 20 50 % Akoline Solution to oil Unchanged Unchanged after 29 Withstand 3
0203-8 MCM, 50 % weeks days in the
Akomed R refrigerator,
MCT but not 4 days,
due to + cholesterol solidification of
(40 mg/g) MCM
57 ACAlO 20 50 % Akoline Solution to oil Unchanged Unchanged after 29 Withstand 3
0203-C MCM, 50 % weeks weeks in the
Akomed R refrigerator
MCT
+ cholesterol
(60 mdgl
58 ACAlO 20 50 % Akoline Solution to oil First signs of Precipitation
0204-1 MCM, 50 % precipitation after a few
Akomed R hours, large star-shaped
MCT crystals after 2 days
+ cholesterol
(1 00 mg/gl
60
61
62
63
64
ACAl0
0206-1
ACAlO
0206-2
ACAlO
0206-3
ACAlO
0206-4
ACAlO
0206-5
20
20
20
20
20
(40 mdgl
30 % Akoline
MCM, 70 %
Akomed R
MCT
+ cholesterol
(60 mg/g)
30 % Akoline
MCM, 70 %
Akomed R
MCT
+ cholesterol
(1 00 mg/gl
15 % Akoline
MCM, 85 %
Akomed R
MCT
+ cholesterol
(40 mdgl
15 % Akoline
MCM, 85 %
Akomed R
MCT
+ cholesterol
(60 mdgl
15 % Akoline
MCM, 85 %
Akomed R
MCT
+ cholesterol
(1 00 mg/gl
Solution to oil
Solution to oil
Solution to oil
Solution to oil
Solution to oil
Unchanged
First signs of
precipitation after about
30 min
Precipitation
Unchanged after 1 days
Signs of precipitation
directly after evaporation
Unchanged after 7
weeks
Precipitation
Precipitation
Precipitation
Precipitation
Withstand 6
days in the
refrigerator,
but had
precipitated
after about 2
weeks without
concomitant
solidification of
MCM
Massive
precipitation
after 28 weeks
66
67
68
ACAlO
02 1 3-C
ACAlO
02 1 3-8
ACAlO
0301-1
20
20
25
(40 mdgl
100 %
Akomed R
MCT
+ cholesterol
(60 mdgl
100 %
Akomed R
MCT
+ cholesterol
(80 mdgl
50 % Akoline
MCM, 50 %
Akomed R
MCT
+ cholesterol
(50 mdg)
Solution to oil
Solution to oil
Solution to oil
Precipitation in the whole
sample
Precipitation directly after
evaporation
Unchanged
Precipitation in the
whole sample
Precipitation in the
whole sample
Unchanged A portion of
the sample
was placed in
the
refrigerator:
partial
solidification
after 1 day,
but clear
solution after
melting at RT.
After another 1
day in the
fridge the
sample started
to precipitate
Massive
precipitation
after 25 weeks
70
71
72
ACAlO
030 1-3
ACAlO
030 1-4
ACAlO
030 1-5
25
30
30
+ cholesterol
(75 mdg)
50 % Akoline
MCM, 50 %
Akomed R
MCT
+ cholesterol
(1 00 mg/g)
50 % Akoline
MCM, 50 %
Akomed R
MCT
+ cholesterol
(60 mdg)
50 % Akoline
MCM, 50 %
Akomed R
MCT
+ cholesterol
(90 mdg)
Solution to oil
Solution to oil
Solution to oil
Very small amount of
needle-shaped crystals
after 1 day
Unchanged after 3 days,
slight tendency to
precipitation after 5 days
Unchanged after 3 days,
precipitation after 5 days
Precipitation
Precipitation
Precipitation
partial
solidification,
but clear
solution after
melting at RT.
The sample
withstood
repeated
cycles during
several weeks
without
precipitation!
A portion of
the sample
was placed in
the
refrigerator:
partial
solidification
and very slight
precipitation
after melting at
RT
A portion of
the sample
was placed in
the
refrigerator:
partial
solidification
and slight
precipitation
after melting at
RT
A portion of
the sample
was placed in
the
refrigerator:
partial
solidification
and substantial
precipitation
after melting at
RT
Example 76
74
75
[00078] The objective of the study was to investigate the comparative
pharmacokinetics in plasma of a 3-beta-hydroxy-5-alpha-pregnan-20-onef ormulation
comprising sesame oil and cholesterol following subcutaneous administration to New
Zealand White rabbits. Two groups of three female rabbits each received single doses
1 mg/kg (formulation as in example 7) or 5 mg/kg (formulation as in example 10).
Following subcutaneous injection into the dorsal neck region of the animals, blood
samples were taken at 0.25, 0.5, 1, 2, 4, 6, 8, 1 2 and 24 hours post dose.
Concentrations of 3-beta-hydroxy-5-alpha-pregnan-20-one in plasma were measured
by a validated LC-MS/MS method. Data are presented in Figure 1 , and show mean
plasma concentration (ng/mL) of 3-beta-hydroxy-5-alpha-pregnan-20-onefo r the two
doses: 1 mg/kg (squares) and 5 mg/kg (circles).
ACAlO
0320
ACAlO
032 1
25
20
(1 20 mg/g)
50 % Akoline
MCM, 50 %
Akomed R
MCT
+ cholesterol
(75 mg/g)
30 % Akoline
MCM, 70 %
Akomed R
MCT
+ cholesterol
(60 mg/g)
discarded)
Solution to oil
Solution to oil
Unchanged
Unchanged
Unchanged
Unchanged
Batch size 100
g
Very slight
precipitation
after 23 weeks
Batch size 100
g
Slight
precipitation
after 23 weeks
Example 77
[00079] Suspensions of 3-beta-hydroxy-5-alpha-pregnan-20-onew ere prepared for
investigation of solubility as follows: cholesterol was first dissolved at room temperature
in sesame oil at 10 and 20 mg/ml, respectively. Suspensions of 3-beta-hydroxy-5-
alpha-pregnan-20-one in sesame oil with different amounts of cholesterol were
prepared on a magnetic stirrer at about 500 rpm using a conventional Teflon coated
stir bar, at room temperature for several days, with occasional cycling to 2-8°C. At this
time, the particles have become substantially smaller. Thereafter, samples of the
respective suspensions were filtered through a 0.2 pm filter and analyzed with regard
to concentration of 3-beta-hydroxy-5-alpha-pregnan-20-one.T he results are presented
in Table 3 and Figure 2 where it can be seen that increasing the amounts of
cholesterol increases the soluble fraction of 3-beta-hydroxy-5-alpha-pregnan-20-one.
Figure 2 shows data for a 3-beta-hydroxy-5-alpha-pregnan-20-onec oncentration of 1 0
mg/g. Figure 3a and 3 b show microscopy pictures taken at 5x enlargement of
suspensions with a 3-beta-hydroxy-5-alpha-pregnan-20-onec oncentration of 1 0 mg/g
and a cholesterol:3-beta-hydroxy-5-alpha-pregnan-20-one ratio of 1 : 1 immediately
upon mixing (3a) and after several days of stirring (3b).
Table 3
Example 78
[00080] Micronized 3-beta-hydroxy-5-alpha-pregnan-20-one (1 0 mg/g) with a
mean particle size of 6 micrometer was suspended in sesame oil with cholesterol (20
mg/g) and stirred as in example 77. Photos were taken immediately upon suspension
(Fig 4a) and after 19 hours of stirring (Fig 4b).
Conclusion
[0008 11 It has been shown that the presence of a sterol such as cholesterol
improves the possibilities to formulate 3-beta-hydroxy-5-alpha-pregnan-20-one in a
pharmaceutical composition comprising acylglycerols, either as an oily solution or an
oily suspension.

CLAIMS
1. A pharmaceutical composition comprising 3-beta-hydroxy-5-alpha-pregnan-
20-one, at least one sterol or an ester thereof and a mixture of acylglycerols with a
solid fat content of less than about 25% at 25°C and about 0% at 37°C.
2. The pharmaceutical composition according to clam 1 wherein the sterol is
selected from the group consisting of cholesterol, beta-sitosterol, stigmasterol,
brassicasterol and avenasterol.
3. The pharmaceutical composition according to claim 2 wherein the sterol is
cholesterol.
4. The pharmaceutical composition according to any one of claims 1 to 3
wherein the mixture of acylglycerols is a vegetable oil.
5. The pharmaceutical composition according to claim 4 where the vegetable
oil is selected from the group consisting of sesame oil, peanut oil, olive oil, and castor
oil.
6. The pharmaceutical composition according to any one of claims 1 to 3
wherein the mixture of acylglycerols is a medium-chain acylglycerol.
7. The pharmaceutical composition according to any one of claims 1 to 6 in
which 3-beta-hydroxy-5-alpha-pregnan-20-one is essentially dissolved.
8. The pharmaceutical composition according to any one of claims 1 to 6
comprising a suspension of 3-beta-hydroxy-5-alpha-pregnan-20-one.
9. A pharmaceutical composition for parenteral administration according to
any one of claims 1 to 8.
10. A pharmaceutical composition for oral administration according to any one
of claims 1 to 8.
1 1 . A pharmaceutical composition for vaginal administration according to any
one of claims 1 to 8.
12. A pharmaceutical composition for nasal administration according to any
one of claims 1 to 8.
13. A method for preparing a pharmaceutical composition according to claim 7
comprising the steps of
a) dissolving 3-beta-hydroxy-5-alpha-pregnan-20-one in ethanol,
b) adding a mixture of acylglycerols with a solid fat content of less than
about 25% at 25°C and about 0% at 37°C and a sterol or an ester
thereof,
c) mixing until a homogeneous liquid is obtained and
d) evaporating the ethanol.
14. A pharmaceutical composition obtainable by the method according to claim
13.
15. Use of a pharmaceutical composition according to any one of claims 1 to
12 or claim 14 for the treatment or prevention of conditions of the central nervous
system.
16. The use according to claim 15 where the condition of the central nervous
system is selected from the group consisting of epilepsy, menstruation cycle dependant
epilepsy, depression, stress related depression, migraine, tiredness and in particular
stress related tiredness, premenstrual syndrome, premenstrual dysphoric disorder,
menstrual cycle linked mood changes, stress related memory changes, menstrual cycle
linked memory changes, Alzheimer's dementia, menstrual cycle linked difficulties in
concentration, menstrual cycle linked sleep disorders and tiredness, substance abuse,
menstrual cycle linked alcoholism, side effects of oral contraceptives and
postmenopausal therapy or combinations thereof
17. Use of the pharmaceutical composition according to any one of claims 1 to
12 or claim 14 for the termination of steroid induced anaesthesia.

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Patent Number

280053

Indian Patent Application Number

5787/DELNP/2012

PG Journal Number

06/2017

Publication Date

10-Feb-2017

Grant Date

08-Feb-2017

Date of Filing

28-Jun-2012

Name of Patentee

ASARINA PHARMA AB

Applicant Address

Fogdevreten 2 SE-171 65 Solna

Inventors:

#	Inventor's Name	Inventor's Address
1	BÄCKSTRÖM Torbjörn	Sofiehemsvägen 73A, SE-907, 38 Umeå
2	CARLSSON Anders	Sankt Göransgatan 80 4tr SE-112 38 Stockholm

PCT International Classification Number

A61K31/57,A61K31/575,A61K47/44

PCT International Application Number

PCT/SE2011/050036

PCT International Filing date

2011-01-14

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1	61/295,027	2010-01-14	Sweden
2	1050029-6	2010-01-14	Sweden