AU6511599A

AU6511599A - Pregnane glucuronides

Info

Publication number: AU6511599A
Application number: AU65115/99A
Authority: AU
Inventors: Michael David Collini; Christopher Paul Miller; Bach Dinh Tran
Original assignee: American Home Products Corp
Current assignee: Wyeth LLC
Priority date: 1998-10-13
Filing date: 1999-10-07
Publication date: 2000-05-01
Anticipated expiration: 2019-10-07
Also published as: EA200100437A1; TW499435B; CZ20011343A3; EP1121376A1; NO20011825L; WO2000021977A1; CA2346693A1; BR9914521A; AU751708B2; KR20010080102A; HUP0103992A2; IL142441A0; AR020778A1; PL347270A1; CN1330659A; NO20011825D0; ZA200103012B; JP2002527448A; HUP0103992A3

Description

WO 00/21977 PCT/US99/23467 PREGNANE GLUCURONIDES BACKGROUND OF THE INVENTION 5 The use of naturally occurring estrogenic compositions of substantial purity and low toxicity such as PREMARIN (conjugated equine estrogens) has become a preferred medical treatment for alleviating the symptoms of menopausal syndrome, osteoporosis/osteopenia in estrogen deficient women and in other hormone related disorders. The estrogenic components of the naturally occurring 10 estrogenic compositions have been generally identified as sulfate esters of estrone, equilin, equilenin, 17-p-estradiol, dihydroequilenin and 17-p-dihydroequilenin (U.S. Patent 2,834,712). The estrogenic compositions are usually buffered or stabilized with alkali metal salts of organic or inorganic acids at a substantially neutral pH of about 6.5 to 7.5. Urea has also been used as a stabilizer (U.S. 15 3,608,077). The incorporation of antioxidants to stabilize synthetic conjugated estrogens and the failure of pH control with tris(hydroxymethyl)aminomethane (TRIS) to prevent hydrolysis is discussed in U.S. 4,154,820. Two of the compounds described herein, 5c-Pregnane-3p,(20S), 21 triol 20-O-p-glucuronide sodium salt and 5a-Pregnane-3p,20R-diol 20-O-p3 20 glucuronide sodium salt are minor components of PREMARIN (conjugated equine estrogens). DESCRIPTION OF THE INVENTION 25 In accordance with this invention, there is provided 5x-Pregnane-3p,(20S), 21-triol, 20-O-p-glucuronide and 5a-Pregnane-3p,20R-diol, 20-O-p-glucuronide and pharmaceutically acceptable salts thereof which are useful as progestational agents. The preparation of pregnane glucuronides 7 and 12 is shown in schemes I and II, respectively. Additionally, a conversion of the glucuronides to the naturally 30 occurring sodium salt forms 8 and 13 is also shown. Pharmaceutically acceptable salts of 5a(-Pregnane-3p,(20S), 21-triol 20-O-p glucuronide and 5ox-Pregnane-3p,20R-diol 20-O-p-glucuronide are not limited to the naturally occurring form, but also include the alkali metal salts, alkaline earth metal 35 salts, ammonium salts, alkylammonium salts containing 1-6 carbon atoms or dialkylammonium salts containing 1-6 carbon atoms in each alkyl group, and WO 00/21977 PCT/US99/23467 -2 trialkylammonium salts containing 1-6 carbon atoms in each alkyl group as well as any other atom or molecules which have a positive charge. As 5a-Pregnane-3p3,(20S), 21 triol 20-O-p-glucuronide and 5a-Pregnane 5 3p,20R-diol 20-O-p3-glucuronide are minor components of PREMARIN (conjugated equine estrogens), this invention also provides 5x-Pregnane-3p3,(20S), 21-triol 20-0 p-glucuronide and 5a-Pregnane-3p,20R-diol 20-O-p-glucuronide and their pharmaceutically acceptable salts in greater than 1 percent purity. 10 This invention also provides compounds consisting essentially of 5a Pregnane-3p3,(20S), 21-triol 20-O-p-glucuronide or a pharmaceutically acceptable salt thereof or 5a-Pregnane-3p,20R-diol 20-O-p3-glucuronide or pharmaceutically acceptable salt thereof. 15 This invention further provides a method of using 5a-Pregnane-3p,(20S), 21 triol glucuronide and 5a-Pregnane-3p3,20R-diol glucuronide or a pharmaceutically acceptable salt of the related glucuronides as progestational agents. The starting materials used in this synthesis are either commercially available 20 or can be prepared using standard chemical methodology. The compounds of this invention can be prepared from readily available starting materials according to the processes in Scheme I, as shown for 5a-Pregnane 3p,(20S), 21-triol 20-O-p-glucuronide (and corresponding monobasic sodium salt) or 25 according to the process in Scheme II, as shown for the synthesis of 5a-Pregnane 3P3,20R-diol 20-O-p-glucuronide (and corresponding monobasic sodium salt). In Scheme I, pregnenolone 3-acetate 1 was used as the starting material. Reaction of 1 with lead tetraacetate in acetic acid according to the procedure 30 described by Purdy, et al, Journal of Medicinal Chemistry 33(6), 1572-1581 (1990) yields 2. Hydrogenation of 2 over PtO 2 in acetic acid yields compounds 3 and 4 in an approximate ratio of 3:1. Compound 3 is subsequently heated with excess equivalents of the acetobromo glucuronic acid methyl ester 5. The protected glucuronide is subsequently saponified with LiOH and reprotonated with acid to yield 35 the desired 20-glucuronic acid conjugate 7. The free acid can be titrated with a slight excess of NaOH solution which generates the monosodium salt 8.

WO 00/21977 PCT/US99/23467 AC Scheme I 0 Pb(OAc) 4 , AcOH H. Pregnenolone OAC2H 1 OH OAc Pt0 2 , AcOH+ H-2, 40 psi0 O'JO 3 4 Ac ~ OM OO~c 5 A H OH ,C 02N5H $w'

-

MH 6 HO OH AC5eOaNaO HOM Ho 8 WO 00/21977 PCT/US99/23467 -4 In Scheme II, compound 10 was reacted with acetobromo glucuronic acid methyl ester 5 and Ag 2

CO

3 to yield the protected conjugate 11. This material was subsequently saponified and then protonated with acid to yield the deprotected glucuronic acid derivative 12. This material was then titrated with NaOH to yield the 5 sodium glucuronate 13. Scheme II OH Ag 2 CO C4pH3 B ACO2PH3 O AC HO Ac OA OAc H 5 11 10 LiOH THF/MeOH/H2O 4h- #kCOZ-%+ Q4 O %%CO2H . NaOH . O3H 'MeOH OH HO &HO H H 12 13 10 The compounds of this invention are progestational agents, and are therefore useful as oral contraceptives (male and female), in hormone replacement therapy (particularly when combined with an estrogen), in the treatment of endometriosis, luteal phase defects, benign breast and prostatic diseases and prostatic and endometrial cancers. The compounds of this invention are also useful in protecting 15 against epileptic seizures, in cognition enhancement, in treating Alzheimer's disease, dementias, vasomotor symptoms related to menopause, and other central nervous system disorders The compounds of this invention are further useful in stimulating erythropoieses.

WO 00/21977 PCT/US99/23467 -5 The compounds of this invention can be used alone as a sole therapeutic agent or can be used in combination with other agents, such as other estrogens, progestins, or androgens. 5 The compounds of this invention can be formulated neat or with a pharmaceutical carrier for administration, the proportion of which is determined by the solubility and chemical nature of the compound, chosen route of administration and standard pharmacological practice. The pharmaceutical carrier may be solid or liquid. 10 A solid carrier can include one or more substances which may also act as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, compression aids, binders or tablet-disintegrating agents; it can also be an encapsulating material. In powders, the carrier is a finely divided solid which is in 15 admixture with the finely divided active ingredient. In tablets, the active ingredient is mixed with a carrier having the necessary compression properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain up to 99% of the active ingredient. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, 20 starch, gelatin, cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins. Liquid carriers are used in preparing solutions, suspensions, emulsions, syrups, elixirs and pressurized compositions. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic 25 solvent, a mixture of both or pharmaceutically acceptable oils or fats. The liquid carrier can contain other suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers or osmo-regulators. Suitable examples of liquid carriers for oral and parenteral administration include 30 water (partially containing additives as above, e.g. cellulose derivatives, preferably sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, e.g. glycols) and their derivatives, lethicins, and oils (e.g. fractionated coconut oil and arachis oil). For parenteral administration, the carrier can also be an oily ester such as ethyl oleate and isopropyl myristate. Sterile liquid 35 carriers are useful in sterile liquid form compositions for parenteral administration.

WO 00/21977 PCT/US99/23467 -6 The liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellant. Liquid pharmaceutical compositions which are sterile solutions or suspensions can be utilized by, for example, intramuscular, intraperitoneal or subcutaneous 5 injection. Sterile solutions can also be administered intravenously. The compounds of this invention can also be administered orally either in liquid or solid composition form. The compounds of this invention may be administered rectally or vaginally in the form of a conventional suppository. For administration by intranasal or 10 intrabronchial inhalation or insufflation, the compounds of this invention may be formulated into an aqueous or partially aqueous solution, which can then be utilized in the form of an aerosol. The compounds of this invention may also be administered transdermally through the use of a transdermal patch containing the active compound and a carrier that is inert to the active compound, is non toxic to the skin, and allows 15 delivery of the agent for systemic absorption into the blood stream via the skin. The carrier may take any number of forms such as creams and ointments, pastes, gels, and occlusive devices. The creams and ointments may be viscous liquid or semisolid emulsions of either the oil-in-water or water-in-oil type. Pastes comprised of absorptive powders dispersed in petroleum or hydrophilic petroleum containing the 20 active ingredient may also be suitable. A variety of occlusive devices may be used to release the active ingredient into the blood stream such as a semipermeable membrane covering a reservoir containing the active ingredient with or without a carrier, or a matrix containing the active ingredient. Other occlusive devices are known in the literature. 25 The dosage requirements vary with the particular compositions employed, the route of administration, the severity of the symptoms presented and the particular subject being treated. Based on the results obtained in the standard pharmacological test procedures, projected daily dosages of active compound would be 0.02 ptg/kg 750 fig/kg. Treatment will generally be initiated with small dosages less than the 30 optimum dose of the compound. Thereafter the dosage is increased until the optimum effect under the circumstances is reached; precise dosages for oral, parenteral, nasal, or intrabronchial administration will be determined by the administering physician based on experience with the individual subject treated. Preferably, the pharmaceutical composition is in unit dosage form, e.g. as tablets or 35 capsules. In such form, the composition is sub-divided in unit dose containing appropriate quantities of the active ingredient; the unit dosage forms can be packaged WO 00/21977 PCT/US99/23467 -7 compositions, for example, packaged powders, vials, ampoules, pre filled syringes or sachets containing liquids. The unit dosage form can be, for example, a capsule or tablet itself, or it can be the appropriate number of any such compositions in package form. 5 The following provides the preparation of representative compounds of this invention. Example 1 10 Pregnane-3p,20R,21-triol 3.21-diacetate 3 Compound 2 (6.0 g, 14.4 mmol) in 0.2 L AcOH was treated with PtO, (1.75 g, 7.7 mmol) and the solution was hydrogenated under 40 PSI of H,. After 18 h, the catalyst was filtered and the crude reaction mixture was concentrated and chromatographed on silica gel using EtOAc/hexanes (1:4) to yield the two products 15 (3 and 4). The first product to elute was the major (desired) material 3 which was isolated as 2.40 g of a white solid: Mp = 164 - 166 0 C; 'H NMR (CDC13) 4.75 - 4.60 (m, 1 H), 4.16 (dd, 1 H, J = 11.4 Hz, 2.2 Hz), 3.94 - 3.87 (m, 1 H), 3.81 - 3.73 (m, 1 H), 2.10 (s, 3 H), 2.10 - 2.05 (m, 1 H), 2.02 (s, 3 H), 1.84 (d, 1 H, J = 5.2 Hz), 1.82 0.88 (m, 21 H), 0.83 (s, 3 H), 0.76 (s, 3 H), 0.68 (dt, 1 H, J = 10.6 Hz); MS (+ESI) 20 421 (M+H)*; IR (KBr) 3520, 2920, 2880, 1740, 1710 cm'. Example 2 Pre2nane-3p.20R,21-triol 3,20-diacetate 4 The second product 4 to elute was the minor product which was isolated as 25 0.73 g of a white solid: Mp = 189 - 191OC; 'H NMR (DMSO) 4.91 (dq, 1 H, J = 5.5 Hz, 2.2 Hz), 4.74 - 4.63 (m, 1 H), 3.81 - 3.74 (m, 1 H), 3.57 - 3.49 (m, 1 H), 2.09 (s, 3 H), 2.02 (s, 3 H), 1.91 (dd, 1 H, J = 7.2 Hz, 5.0 Hz), 1.85 - 0.83 (m, 22 H), 0.82 (s, 3 H), 0.72 - 0.61 (m, 1 H), 0.65 (s, 3 H); MS El 420 (M+); IR (KBr) 3410, 2920, 1730, 1700 cm'. 30 WO 00/21977 PCT/US99/23467 -8 Example 3 2.3.4-O-Triacetyl -1-O-(3p,21-diacetoxy-5a-pregnan-20S-vl)---D glucuronic acid methyl ester 6 Compound 3 (6.6 g, 15.7 mmol) was dissolved in CHCI 3 (125 mL) and treated 5 with Ag 2

CO

3 (4.3 g, 15.7 mmol) and the glucuronyl bromide 5 [21085-72-3] (6.2 g, 15.7 mmol). After refluxing for 1.5 h, an additional 0.5 eq of Ag 2

CO

3 and 0.5 eq of the glucuronyl bromide were added. After an additional 1.5 h this addition of reagents was repeated and after another 1.5 h it was repeated once more. The reaction was allowed to reflux for a total of 5.5 h. The reaction was allowed to cool 10 down and the inorganic salts were filtered off. The filtrate was concentrated and chromatographed on silica gel (3:7, EtOAc/hexanes) and the fractions containing product were concentrated and rechromatographed under the same conditions to yield 4.1 g of 6 which was triturated with MeOH to give 3.05 g of 6 as a white solid: Mp = 180 - 183oC; MS (APCI) 754 (M+ NH4+); 'H NMR (DMSO) 5.36 (t, 1 H, J = 9.6 15 Hz), 5.06 (d, 1 H, J = 8.0 Hz), 4.91 (t, 1 H, J= 9.8 Hz), 4.70 (dd, 1 H, J = 9.5 Hz, 8.1 Hz), 4.63 - 4.50 (m, 1 H), 4.47 (d, 1 H, J = 10.0 Hz), 4.16 (d, 1 H, J = 11.3 Hz), 3.92 - 3.75 (m, 2 H), 3.63 (s, 3 H), 2.16 - 2.07 (m, 1 H), 2.04 (s, 3 H), 1.99 (s, 3 H), 1.97 (s, 3 H), 1.95 (s, 3 H), 1.93 (s, 3 H), 1.78 - 1.63 (m, 3 H), 1.62 - 1.39 (m, 6 H), 1.36 1.10 (m, 8H), 1.06 - 0.85 (m, 5 H), 0.79 (s, 3 H), 0.68 (s, 3 H). 20 Example 4 5c-Pregnane -3p.20S,21-triol 20-O-D-D-glucuronide 7 Compound 6 (3.76 g, 5.1 mmol) was dissolved in THF (25 mL) and treated with a solution consisting of LiOH (1.35 g, 56.1 mmol) in H20 (13 mL). MeOH (4 25 mL) was added to this solution and the reaction was heated to 75oC for 2 h. The solution was then cooled and concentrated. The aqueous residue was taken up with an additional 15 mL of H20. To this residue was then added a 2N HC1 solution (43 mL). Solid formation was induced by scratching the glass. Filtration yielded 2.6 g of 7 as a white solid: 30 Mp = 231 - 235oC; " 3 C NMR (75 MHz, MeOD) (C=O missing), 102.6, 82.1, 77.6, 76.8, 75.3, 73.1, 71.9, 63.6, 57.6, 56.1, 51.5, 49.9, 46.3, 43.6, 40.3, 39.0, 38.3, 36.9, 36.7, 33.5, 32.2, 30.0, 25.9, 25.4, 22.3, 12.8, 11.9; 'H NMR (300 MHz, MeOD) 4.54 WO 00/21977 PCT/US99/23467 -9 (d, 1 H, J = 7.7 Hz), 3.82 - 3.71 (m, 3 H), 3.59 - 3.22 (m, 5 H), 2.30 (d, 1 H, J = 12.6 Hz), 1.75 - 1.66 (m, 6 H), 1.53 - 1.18 (m, 9 H), 1.17 - 0.88 (m, 6 H), 0.83 (s, 3 H), 0.78 (s, 3 H), 0.70 - 0.58 (m, 1 H); MS (-)ESI 511 (M-H); IR (KBr) 3410, 2910, 2830, 1730, 1700 cm 1. 5 Example 5 5(x-Pregnane-3p3.20S.21-triol 20-O-p3-D- 2lucuronide sodium salt 8 Compound 7 (1.57 g, 1.7 mmol) was suspended in MeOH (20 mL) and treated with an aqueous solution of NaOH (6.4 mL, 0.5 N) and stirred for 5 minutes which 10 allowed all of the starting material to go into solution. The solution was then mixed with 0.65 g of product from a previous run and the combination stripped onto silica gel and column chromatographed on silica gel (MeOH:CHC1,, 4:6 then 5:5). The product was then triturated once with a 1:1 mixture of MeOH and Dioxane (total volume equal 60 mL) to yield 1.57 g of 8 as a white solid: Mp = 240 - 244oC; 13C 15 NMR (75 MHz, DMSO) (C=O missing), 100.4, 79.8, 76.7, 73.9, 73.7, 72.2, 69.3, 66.3, 62.0, 55.6, 54.0, 49.8, 44.3, 41.9, 38.1, 36.6, 35.1, 35.0, 31.8, 31.3, 28.4, 24.4, 23.9, 20.8, 12.1, 11.4; IR (KBr) 3400 (H 2 0), 2920, 2870, 1610 cm'. Example 6 20 Pregnane-3p,20S.21-triol 9 Compound 3 (1.6 g, 3.8 mmol) was dissolved in THF (8 mL). A solution of LiOH (0.27 g, 11.4 mmol) in 3 mL of H,O was added. In order to make the solution one phase, 1 mL of MeOH was added to the reaction mixture. After 30 minutes at reflux, the reaction mixture was treated with 1.1 mL of AcOH and partitioned 25 between an organic layer consisting of 100 mL EtOAc, 20 mL CH 2 C1 2 , 10 mL MeOH, and 60 mL H 2 0. The organic layer was then washed with brine and dried over MgSO 4 . The solution was concentrated and the resulting solid triturated with ether to yield 1.1 g of 9 as a white solid: Mp = 210 - 212oC; 'H NMR (DMSO) (3 OH protons missing) 4.43 (d, 1 H, J = 4.6 Hz), 4.31 (t, 1 H, J = 5.5 Hz), 4.05 (d, 1 H, 30 J = 5.2 Hz), 3.16 - 3.05 (m, 1 H), 2.10 (d, 1 H, J = 12.4 Hz), 1.65 - 0.78 (m, 21 H), 0.75 (s, 3 H), 0.68 (s, 3 H), 0.65 - 0.54 (m, 1 H); MS El 336 M+; IR (KBr) 3400, 2930, 2880 cm'.

WO 00/21977 PCT/US99/23467 -10 Example 7 1-O-( 3 3-Acetoxy-pregn-20-vl)-2,3,4-triacetyl-B-D-elucuronic acid methyl ester 11 5u-Pregnan-33, 203-diol 3-acetate 10 (3.0g, 11.0 mmol), glucuronyl bromide 5 5 [21085-72-3] (5.5g, 13.8 mmol) and Ag 2

CO

3 (4.5g, 16.1 mmol) were stirred in toluene (40 mL) at rt (flask protected from light with aluminum foil). After 18 h the reaction was diluted with CH 2 C 2 , filtered, concentrated and chromatographed on silica gel (EtOAc:Hex, 2:8 then EtOAc:Hex, 4:6) to yield a solid which was triturated with MeOH to give 3.0 g of 11 as a white solid: 10 Mp = 248 - 251oC; 'H NMR (DMSO) 5.33 (t, 1 H, J = 9.6 Hz), 4.95 - 4.88 (m, 2 H), 4.68 (dd, 1 H, J = 9.5 Hz, 8.1 Hz), 4.62 - 4.49 (m, 1 H), 4.44 (d, 1 H, J = 10.0 Hz), 3.71 - 3.65 (m, 1 H), 3.63 (s, 3 H), 2.11 (d, 1 H, J = 12.6 Hz), 1.98 (s, 3 H), 1.97 (s, 3 H), 1.96 (s, 3 H), 1.95 (s, 3 H), 1.77 - 1.64 (m, 2 H), 1.63 - 1.10 (m, 14 H), 0.99 (d, 3 H, J = 5.8 Hz), 0.95 - 0.83 (m, 5 H), 0.78 (s, 3 H), 0.66 (s, 3 H), 0.65 - 0.58 (m, 1 H); 15 IR (KBr) 2930, 2900, 2830, 1750, 1730 cm'; MS (+)ESI 696 (M+NH 4 ) . Example 8 5a-Pregnane-313,20R-diol 20-O-3-D-2lucuronide 12 Compound 11 (2.5 g, 3.7 mmol) was dissolved in a solution of THF (25 mL) 20 and MeOH (15 mL) and treated with a solution of LiOH (0.9g, 37.5 mmol) in H 2 0 (10 mL). The reaction was heated at reflux for 1 h. The reaction was allowed to cool to room temperature and concentrated. The product was taken up in water (20 mL) and acidified with aqueous 2 N HC1. A precipitate formed which was filtered yielding 1.5 g of 12 as a white solid: Mp = 255 - 260; 'H NMR (DMSO) (3 H 25 buried) 4.98 (d, 1 H, J = 3.8 Hz), 4.91 (d, 1 H, J = 4.6 Hz), 4.42 (br s, 1 H), 4.25 (d, 1 H, J = 7.7 Hz), 3.71 - 3.62 (m, 1 H), 3.58 (d, 1 H, J = 9.7 Hz), 3.23 - 3.12 (m, 1 H), 2.96 - 2.87 (m, 1 H), 2.15 (d, 1 H, J = 12.3 Hz), 1.67 - 1.46 (m, 5 H), 1.45 - 0.80 (m, 17 H), 1.01 (d, 3 H, J = 5.8 Hz), 0.74 (s, 3 H), 0.65 (s, 3 H), 0.65 - 0.52 (m, 1 H); IR (KBr) 3520, 3400, 2920, 2820, 2800, 1710 cm-'; MS (-) ESI 495 (M-H)-. 30 WO 00/21977 PCT/US99/23467 -11 Example 9 5a-Pregnane-3p,20R-diol 20-O-p-D-glucuronide sodium salt 13 A suspension of Compound 12 (1.5 g, 3.0 mmol) in MeOH (25 mL) and treated with an aqueous solution of NaOH (0.5 N, 6.0 mL, 3.0 mmol) and stirred for 5 5 minutes during which time everything went into solution. The solution was concentrated to dryness and the residue was triturated with EtOH to obtain 1.1 g of 13 as a white solid: Mp = 223 - 226 0 C (dec); 'H NMR (DMSO) 7.26 (s, 1 H), 4.76 (d, 1 H, J = 3.6 Hz), 4.67 (d, 1 H, J = 3.6 Hz), 4.43 (d, 1 H, J = approx 4 Hz), 4.12 (d, 1 H, J = 7.6 Hz), 10 3.78 - 3.71 (m, 1 H), 3.45 - 3.35 (m, 1 H), 3.17 - 3.03 (m, 3 H), 3.38 - 3.29 (m, 1 H), 2.17 (d, 1 H, J = 12.4 Hz), 1.67 - 1.49 (m, 5 H), 1.45 - 0.80 (m, 17 H), 0.99 (d, 3 H, J = 5.9 Hz), 0.76 (s, 3 H), 0.75 (s, 3 H), 0.65 - 0.53 (m, 1 H); IR (KBr) 3400, 2920, 2830, 1610 cm'; MS (-) ESI 495 (M-H).

Claims

1. A compound which is 5a-pregnane-3p,20S,21-triol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof. 5 2. The compound of claim 1, wherein the pharmaceutically acceptable salt of the

20-glucuronide is an alkali metal salt, alkaline earth metal salt, ammonium salt, alkylammonium salt containing 1-6 carbon atoms, or dialkylammonium salt containing 1-6 carbon atoms in each alkyl group, or trialkylammonium salt containing 1-6 carbon atoms in each alkyl group. 10 3. A compound which is 5a-pregnane-3p,20R-diol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof. 4. The compounds of claim 3 wherein the pharmaceutically acceptable salt of the 15 20-glucuronide is an alkali metal salt, alkaline earth metal salt, ammonium salt, alkylammonium salt containing 1-6 carbon atoms, or dialkylammonium salt containing 1-6 carbon atoms in each alkyl group, or trialkylammonium salt containing 1-6 carbon atoms in each alkyl group. 20 5. 5a-pregnane-3p,20S,21-triol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof, which is at least 1 percent pure. 6. 5a-pregnane-3p,20R-diol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof which is at least 1 percent pure. 25 7. A compound consisting essentially of 5a-pregnane -3p,20S,21-triol 20-O-p D-glucuronide or a pharmaceutically acceptable salt thereof. 8. A compound consisting essentially of 5a-pregnane-3p,20R-diol 20-O-p-D 30 glucuronide or a pharmaceutically acceptable salt thereof. 9. A compound consisting essentially of 5a-pregnane-3p,20S,21-triol 20-O-p-D glucuronide sodium salt. WO 00/21977 PCT/US99/23467 - 13 10. A compound consisting essentially of 5x-pregnane-3p,20R-diol 20-O-p-D glucuronide sodium salt. 11. A pharmaceutical composition which comprises 5ct-pregnane-3p,20S,21-triol 5 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof and a pharmaceutical carrier. 12. A pharmaceutical composition which comprises 5(-pregnane-3p,20R-diol 20 O-p-D-glucuronide or a pharmaceutically acceptable salt thereof and a 10 pharmaceutical carrier. 13. A pharmaceutical composition according to claim 11 or 12 which further comprises other agents selected from estrogens, progestogens and androgens. 15 14. A pharmaceutical composition according to claim 12 which consists essentially of 5a-pregnane-3p,20S,21-triol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof and at least one pharmaceutical carrier. 15. A pharmaceutical composition according to claim 12 which consists 20 essentially of 5a-pregnane-3p,20R-diol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof and at least one pharmaceutical carrier. 16. A pharmaceutical composition which comprises greater than 1% w/w or w/v of 5x-pregnane-3p,20S,21-triol 20-O-p-D-glucuronide or a pharmaceutically 25 acceptable salt thereof, and at least one pharmaceutical carrier. 17. A pharmaceutical composition which comprises greater than 1% w/w or w/v of 5a-pregnane-3p,20R-diol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof, and at least one a pharmaceutical carrier. 30 18. 5a-pregnane-3p,20S,21-triol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof, for use in the treatment of mammals. 19. 5a-pregnane-3p,20R-diol 20-O-p-D-glucuronide or a pharmaceutically 35 acceptable salt thereof, for use in the treatment of mammals. WO 00/21977 PCT/US99/23467 -14 20. 5x-pregnane-3p,20S,21-triol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof, for use as a progestational agent.

21. 5u-pregnane-3p,20R-diol 20-O-p-D-glucuronide or a pharmaceutically 5 acceptable salt thereof, for use as a progestational agent.

22. 5a-pregnane-3p,20S,21-triol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof, for use in treating or inhibiting cancers, central nervous system disorders, dementias, or alzheimer's disease in a mammal in need thereof. 10

23. 5a-pregnane-3p,20R-diol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof, for use in treating or inhibiting cancers, central nervous system disorders, dementias, or alzheimer's disease in a mammal in need thereof. 15 24. Use of 5a-pregnane-3p,20S,21-triol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use as a progestational agent or in treating or inhibiting cancers, central nervous system disorders, dementias, or alzheimer's disease or in stimulating erythropoieses in a mammal in need thereof. 20

25. Use of 5(-pregnane-3p,20R-diol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use as a progestational agent or in treating or inhibiting cancers, central nervous system disorders, dementias, or alzheimer's disease or in stimulating erythropoieses in a 25 mammal in need thereof.

26. A method of providing progestational therapy to a mammal in need thereof which comprises administering a progestationally effective amount of 5a-pregnane 3p,20S,21-triol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof 30 to said mammal.

27. A method of providing progestational therapy to a mammal in need thereof which comprises administering a progestationally effective amount of 5a-pregnane 3p,20R-diol 20-O-p-D-glucuronide or a pharmaceutically acceptable salt thereof to 35 said mammal. WO 00/21977 PCT/US99/23467 - 15

28. A method of treating or inhibiting cancers, central nervous system disorders, dementias, or alzheimer's disease in a mammal in need thereof, which comprises administering an effective amount of 5ac-pregnane-313,20S,21-triol 20-O-P3-D glucuronide or a pharmaceutically acceptable salt thereof, to said mammal. 5

29. A method of treating or inhibiting cancers, central nervous system disorders, dementias, or alzheimer's disease in a mammal in need thereof, which comprises administering an effective amount of 5a-pregnane-3p,20R-diol 20-O-p3-D glucuronide or a pharmaceutically acceptable salt thereof, to said mammal. 10

30. Process for the preparation of 5u-pregnane-3p3,(20S), 21-triol, 20-O-p3 glucuronide or a pharmaceutically acceptable salt thereof, which comprises reacting pregnenolone 3-acetate with lead tetraacetate in acetic acid to give the compound of formula 2 : 15 OAc O0 0 2 which is then hydrogenated over PtO 2 in acetic acid to give compound 3: Ac OH 0 0 20 3 which is subsequently heated with excess equivalents of the compound 5 WO 00/21977 PCT/US99/23467 - 16 Br O \CO 2 Me AcO . OAc QAc 5 to give the compound of formula 6 : CO 2 Me 10 OAc which ~ ~ ~ ~ ~ ~~- A.pie ecin opud1 10~ 3p,(20S), 21-triol, 20-O-p-glucuronide, and optionally this compound can be converted to the pharmaceutically acceptable salt thereof. 10

31. Process for the preparation of 5x-pregnane-3p,20R-diol, 20-O-p-glucuronide which comprises reacting compound 10 : OH 0 H 10 with a compound of formula 5: Br O OCH3 AcO OAc 3Ac 15 5 WO 00/21977 PCT/US99/23467 - 17 and Ag 2 CO 3 to yield a compound of formula 11: CO 2 CH 3 I 0 . OAC 0 JJW' ACc 0 0 H 11 5 which is subsequently saponified and reprotonated with acid to 5c-pregnane-3p3,20R diol, 20-O-p-glucuronide, according to claim and optionally this compound can be converted to the pharmaceutically acceptable salt thereof.

32. Process for the preparation of the monosodium salt of 5a-pregnane-3p3,(20S), 10 21-triol, 20-O-p3-glucuronide which comprises titrating with a slight excess of NaOH solution.

33. Process for the preparation of the monosodium salt of 5a-pregnane-3p3,20R diol, 20-O-p3-glucuronide which comprises titrating with a slight excess of NaOH 15 solution.