ZA200103012B

ZA200103012B - Pregnane glucoronides.

Info

Publication number: ZA200103012B
Application number: ZA200103012A
Authority: ZA
Inventors: Christopher Paul Miller; Michael David Collini; Bach Dinh Tran
Original assignee: Wyeth Corp
Priority date: 1998-10-13
Filing date: 2001-04-11
Publication date: 2002-07-11
Also published as: CA2346693A1; NO20011825D0; AU751708B2; AU6511599A; EP1121376A1; IL142441A0; PL347270A1; EA200100437A1; HUP0103992A2; HUP0103992A3; NO20011825L; AR020778A1; TW499435B; CN1330659A; BR9914521A; WO2000021977A1; KR20010080102A; CZ20011343A3; JP2002527448A

Description

= bod! WO 00/21977 PCT/US99/23467

PREGNANE GLUCURONIDES

BACKGROUND OF THE INVENTION

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 estrogenic compositions have been generally identified as sulfate esters of estrone, equilin, equilenin, 17-p-estradiol, dihydroequilenin and 17-B-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. 3,608,077). The incorporation of antioxidants to stabilize synthetic conjugated estrogens and the failure of pH control with tris(hydroxymethyDaminomethane (TRIS) to prevent hydrolysis is discussed in U.S. 4,154,820.

Two of the compounds described herein, Sa-Pregnane-3p,(20S), 21 triol 20-O-B-glucuronide sodium salt and Sa-Pregnane-3B,20R-diol 20-O-p- glucuronide sodium salt are minor components of PREMARIN (conjugated equine estrogens).

DESCRIPTION OF THE INVENTION

Ii accordance with this invention, whore is provided So-licgnanc-2p,{205), 21-triol, 20-O-B-glucuronide and Sa-Pregnane-3p,20R-diol, 20-O-B-glucuronide and pharmaceutically acceptable salts thereof which arc usctul as progestational agents.

The preparation of pregnane glucuronides 7 and 12 is shown in schemes I and If, respectively. Additionally, a conversion of the glucuronides to the naturally occurring sodium salt forms 8 and 13 is also shown.

Pharmaceutically acceptable salts of Sa-Pregnane-3p,(20S), 21-triol 20-O-- glucuronide and Sa-Pregnane-3B,20R-diol 20-O-B-glucuronide are not limited to the naturally occurring form, but also include the alkali metal salts, alkaline earth metal salts, ammonium salts, alkylammonium salts containing 1-6 carbon atoms or dialkylammonium salts containing 1-6 carbon atoms in each alkyl group, and 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 5Sa-Pregnane-3B,(20S), 21 triol 20-O-B-glucuronide and Sa-Pregnane- 3B,20R-diol 20-O-B-glucuronide are minor components of PREMARIN (conjugated equine estrogens), this invention also provides 5a-Pregnane-38,(208S), 21-triol 20-O-

B-glucuronide and Sa-Pregnane-3B,20R-diol 20-O-B-glucuronide and their pharmaceutically acceptable salts in grcater than 1 percent purity.

This invention also provides compounds consisting essentially of 5a-

Pregnane-3,(20S), 21-triol 20-O-B-glucuronide or a pharmaceutically acceptable salt thereof or S5a-Pregnane-3p,20R-diol 20-O-B-glucuronide or pharmaceutically acceptable salt thereof.

This invention further provides a method of using Sa-Pregnane-3p,(208S), 21- triol glucuronide and Sa-Pregnane-3B,20R-diol glucuronide or a pharmaceutically acceptable salt of the related glucuronides as progestational agents.

The starting materials used in this synthesis arc either commercially available or can be prepared using standard chemical methodology.

The compounds of this invention can be prepared from readily available starting materials according to the proccsses in Scheme I, as shown for Sa-Pregnane- 3B,(20S), 21-triol 20-O-B-glucuronide (and corresponding monobasic sodium salt) or according to the process in Scheme II, as shown for the synthesis of Sa-Pregnanc- : 3B,20R-diol 20-O-B-glucuronide (and corresponding monobasic sodium salt).

In Scheme 1, pregnenolone 3-acetate 1 was used as the starting material.

Reaction of 1 with lead tetraacetate in acetic acid according to the procedure described by Purdy, et al, Journal of Medicinal Chemistry 33(6), 1572-1581 (1990) yields 2. Hydrogenation of 2 over PtO, 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 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.

- hd WO 00121977 PCT/US99/23467

AC

Scheme {o]

Pb(OAc),. AcOH t —_—

X ’ AL © 0

Pregnenolone AC ht byl 2 oH OAc

P10, ACOH .

Hj. 40 psi x J hi d 3 4

Soom fo Lo

OAC

8 £\CO, M2 { 3s, Ag,CO,. CHC, AD Dac —————t mn

Ac! Y OAc

Onc )

E 6 oH of -

OH

Ea . 1 - LIOH. THF 1,0. MeOH aT A 6 —_—= NTA A ’ Pra) Nall) r hd Y

NZ

H

—7— -

HCOO-Na+ [o, lad

OH

>

HO » 7 MeOH, 1.05 eq Nao} oH

HO' 2

In Scheme II, compound 10 was reacted with acetobromo glucuronic acid methyl ester 5 and Ag,CO, to vield thc 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 sodium glucuronate 13.

Scheme ll

OH

Ag,CO,4 Jos

B ,MCOLH, ond oh

A :

OAc on z AcO' Y OAc Pe :

H OAc = 5 11

LiOH

THFMeOH/H,0

HO i OH NaOH HO H OH oH hot OH

HO = HO i

Ho 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 brcast and prostatic diseases and prostatic and endometrial cancers. The compounds of this invention are also useful in protecting 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.

: hor WO 00/21977 PCT/US99/23467

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.

The compounds of this invention can be formulated neat or with a pharmaccutical 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.

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 cncapsulating material. In powders, the carrier is a finely divided solid which is in 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, 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 253 sulveid, a wuiaiuie ui UOil OF pharinacéuiically atcopiauic ulld wi fails. Ihc yuid 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 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 carriers are useful in sterile liquid form compositions for parenteral administration.

The liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellant.

Liquid pharmaceutical compositions which arc sterile solutions or suspensions can be utilized by, for example, intramuscular, intraperitoneal or subcutaneous 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 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 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 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.

The dosage requirements vary with the particular compositions employed, the ‘ route of administration, the severity of the symptoms presented and the particular subject being trcated. Based on the results obtained in the standard pharmacological test procedures, projected daily dosages of active compound would be 0.02 pg/kg - 750 pg/kg. Treatment will generally be initiated with small dosages less than the 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 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

: Se WO 00721977 PCT/US99/23467 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.

The following provides the preparation of representative compounds of this invention.

Example 1 Pregnane-38.20R.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 (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°C; 'H NMR (CDCl) 4.75 - 4.60 (m, 1H), 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, 3H), 1.84 (d, 1 H, ] = 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) 421 (M+H)"; IR (KBr) 3520, 2920, 2880, 1740, 1710 cm”.

Example 2 . Pregnone-2R 20D 91 -trinl ? J().-diacetate A

The second product 4 to elute was the minor product which was isolated as 0.73 gof a white solid: "Mp =189-191°C;"H NMR (DMSO) 4:91 (dq, I 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, 3H), 2.02 (,3H), 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 EI 420 (M+); IR (KBr) 3410, 2920, 1730, 1700 cm™.

Example 3 methyl ester 6

Compound 3 (6.6 g, 15.7 mmol) was dissolved in CHCL, (125 mL) and treated with Ag,CO, (4.3 g, 15.7 mmol) and the glucuronyl bromide S [21085-72-3] (6.2 g, 15.7 mmol). After refluxing for 1.5 h, an additional 0.5 eq of Ag,CO, and 0.5 eq of the glucuronyl bromide werc 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 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 trituratcd with McOH to give 3.05 g of 6 as a white solid: Mp = 180 - 183°C; MS (APCI) 754 (M+ NH4"); 'H NMR (DMSO) 5.36 (t, 1 H, J = 9.6

Hz),506(d, 1H, 1=80Hz),491(, 1H,J=9.8Hz),4.70 (dd, 1 H, J=9.5 Hz, 8.1

Hz), 4.63 - 4.50 (m, | 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, 3H), 2.16 - 2.07 (m, 1 H), 2.04 (s, 3 H), 1.99 (s, 3 H), 1.97 (s, 3H), 1.95 (s, 3H), 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, 5S H), 0.79 (s, 3 H), 0.68 (s. 3 H).

Example 4

Sa-Pregnane -3p,208.21-triol 20-O-8-D-glucuronide 7 ’

Compound 6 (3.76 g, 5.1 mmol) was dissolved in THF (25 mL) and treated with a solution consisting of L1IOH (1.35 g, 56.1 mmol) in H,0 (13 mL). MeOH (4 ml) was added to this solution and the reaction was heated to 75°C for 2 h. The solution was then cooled and concentrated. The aqueous residue was taken up with an additional 15 mL of H,0. To this residuc was then added a 2N HCI solution (43 mL). Solid formation was induced by scratching the glass. Filtration yielded 2.6 g of 7 as a white solid:

Mp = 231 - 235°C; °C NMR (75 MHz, MeOD) (C=0 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

) ed WO 00/21977 PCT/US99/23467 (d,1H,)=77Hz),3.82-371 (m,3H),3.59-322(m,5H),230(d, 1 H,J=126

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".

Example 5

Sa-Pregnane-3p,208.21-triol 20-O-B-D-glucuronide 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 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:CH,Cl,, 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 whitc solid: Mp = 240 - 244°C; °C

NMR (75 MHz, DMSO) (C=0 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,0), 2920, 2870, 1610 cm".

Example 6 Pregnane-35.208.21-triol 9

Compound 3 {1.6 g, 3.8 mmol) was dissolved in THF (2 mL). A solution of

LOI {027 2, MA mmol in I mL of HO war added In order 10 make the anhation 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 between an organic layer consisting of 100 mL EtOAc, 20 mL CHCl, 10 mL

MeOH, and 60 mL HO. The organic layer was then washed with brine and dried over MgSO,. The solution was concentrated and the resulting solid triturated with ether to yield 1.1 g of 9 as a white solid: Mp = 210 - 212°C; 'H NMR (DMSO) (3

OH protons missing) 4.43 (d, 1 H, J =4.6 Hz), 431 (1, 1 H, J = 5.5 Hz), 4.05 (d, 1 H,

J=5.2Hz),3.16-3.05(m, 1H), 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 EI 336 M+; IR (KBr) 3400, 2930, 2880 cm”.

Example 7 1-O-(3B-Acetoxy-pregn-20-yl )-2.3.4-triacety]-B-D-glucuronic acid methyl ester 11

Sa-Pregnan-3, 20B-diol 3-acetate 10 (3.0g, 11.0 mmol), glucuronyl bromide §{21085-72-3) (5.5g, 13.8 mmol) and Ag,CO, (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,Cl,, 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:

Mp =248 - 251°C; 'H NMR (DMSO) 5.33 (4, 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, 3H), 2.11(d, 1 H, J = 12.6 Hz), 1.98 (s, 3 H), 1.97 (s, 3

H), 1.96 (s, 3H), 1.95 (s, 3H), 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 (in, 5 H), 0.78 (s, 3 H), 0.66 (s, 3H), 0.65 - 0.58 (m, 1 H);

IR (KBr) 2930, 2900, 2830, 1750, 1730 cm™; MS (+)ESI 696 (M+NH,)".

Example 8

Sa-Pregnane-38.20R-diol 20-O-B-D-glucuronide 12

Compound 11 (2.5 g, 3.7 mmol) was dissolved in a solution of THF (25 mL) and MeOH (15 mL) and treated with a solution of LiOH (0.9g, 37.5 mmol) in H,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 HCl. A precipitate formed which was filtered yielding 1.5 g of 12 as a white solid: Mp = 255 - 260°; '"H NMR (DMSO) (3 H buried) 4.98 (d, 1 H,J =3.8Hz),4.91(d, 1H,J=46 Hz), 4.42 (brs, 1 H), 4.25 (d, 1

H,J=7.7Hz),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, 3H), 0.65-0.52 (m, 1 H); IR (KBr) 3520, 3400, 2920, 2820, 2800, 1710 cm™:; MS (-) ESI 495 (M-H).

) taal WO 00/21977 PCT/US99/23467

Example 9

Sa-Pregnane-38.20R-diol 20-O-B-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 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°C (dec); 'H NMR (DMSO) 7.26 (s, 1 H), 4.76 (d, 1 H, J = 3.6 Hz), 467 (d, 1H,J=3.6Hz),443 (d, | H, J = approx 4 Hz), 4.12 (d, 1 H, J = 7.6 Hz), 3.78 -3.71 (m, 1 H), 3.45 -3.35 (m, 1 H), 3.17 - 3.03 (mm, 3 H), 3.38 - 3.29 (m, 1 H), 2.17(d, 1 H,)=12.4 Hz), 1.67 - 1.49 (m, 5H), 1.45- 0.80 (m, 17H), 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 (-) ES1495 (M-H).

Claims

WHAT IS CLAIMED IS:

1. A compound which is 5a-pregnane-3,20S,21-triol 20-O-B-D-glucuronide or - a pharmaceutically acceptable salt thereof. 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.

3. A compound which is Sa-pregnane-3p,20R-diol 20-O-B-D-glucuronide or a pharmaceutically acceptable salt thereof.

4. The compounds of claim 3 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.

5. Sa-pregnane-3B,20S,21-triol 20-O-B-D-glucuronide or a pharmaceutically acceptable salt thereof, which is at least 1 percent pure.

6. 5a-pregnanc-3p,20R-diol 20-O-B-D-glucuronide or a pharmaceutically accentabie salt Inereof wiicn is at least 1 percent pure. : 7: A-compound-consisting essentially of -Sa=pregnane—=3f;208,2 1=triol-20-0-p- - D-glucuronide or a pharmaceutically acceptable salt thereof.

8. A compound consisting essentially of 5a-pregnane-3p,20R-diol 20-O-p-D- glucuronide or a pharmaceutically acceptable salt thereof.

9. A compound consisting essentially of Sa-pregnane-3p,208S,21-triol 20-O-p-D- glucuronide sodium salt.

» PCT/US99/23467

10. A compound consisting essentially of S5a-pregnane-3f,20R-diol 20-O-B-D- glucuronide sodium salt.

11. A pharmaceutical composition which comprises 5a-pregnane-3,208S,21-triol 20- O-B-D-glucuronide or a pharmaceutically acceptable salt thereof and a pharmaceutical carrier.

12. A pharmaceutical composition which comprises Se-pregnane-3(3,20R-diol 20-O-f3- D-glucuronide or a pharmaceutically acceptable salt thereof and a pharmaceutical carrier.

13. A pharmaceutical composition according to claim 11 or 12 which further comprises other agents selected from estrogens, progestogens and androgens.

14. A pharmaceutical composition according to claim 12 which consists essentially of

5o.-pregnane-3,208S,21-triol 20-O-f3-D-glucuronide or apharmaceutically acceptable salt thereof and at least one pharmaceutical carrier.

15. A pharmaceutical composition according to claim 12 which consists essentially of S¢-pregnane-38,20R diol 20-0-8-D-glucuronide or a pharmaceutically acceptable salt thereof and at least one pharmaceuticai carrier.

16. A pharmaceutical composition which comprises greater than 1% w/w or w/v of Sa-pregnane-3p,20S,21-triol 20-O-B-D-glucuronide or apharmaceutically acceptable salt thereof, and at least one pharmaceutical carrier.

17. A pharmaceutical composition which comprises greater than 1% w/w or w/v of Sa-pregnane-3p,20R-diol 20-O-B-D-glucuronide or a pharmaceutically acceptable salt thereof, and at least one a pharmaceutical carrier. AMENDED SHEET af PCT/US99/23467

18. A substance or composition for use in a method for the treatment of mammals, said substance or composition comprising Sa-pregnane-3f,20S,21-triol 20-O-3-D-glucuronide or a pharmaceutically acceptable salt thereof, and said method comprising administering said substance or composition.

19. A substance or composition for use in a method for the treatment of mammals, said substance or composition comprising Sa-pregnane-3,20R-diol 20-O-B-D-glucuronide or a pharmaceutically acceptable salt thereof, and said method comprising administering said substance or composition.

20. A substance or composition for use in a method of treatment, as a progestational agent, said substance or composition comprising 5a-pregnane-30,208S,21-triol 20-O-3-D- glucuronide or a pharmaceutically acceptable salt thereof, and said method comprising administering said substance or composition.

21. A substance or composition for use in a method of treatment, as a progestational agent, said substance or composition comprising Sa-pregnane-3f3,20R-diol 20-O-3-D- glucuronide or a pharmaceutically acceptable salt thereof, and said method comprising adminisiering said substance of composition.

22. ‘A-substance-or-composition-for-use-in-a-method—for-the-treating-or-inhibiting- cancers, central nervous system disorders, dementias, or alzheimer's disease in a mammal, said substance or composition comprising 5o-pregnane-3p,208S,21-triol 20-O-B-D- glucuronide or a pharmaceutically acceptable salt thereof, and said method comprising administering said substance or composition. AMENDED SHEET

- PCT/US99/23467

23. A substance or composition for use in a method for the treating or inhibiting cancers, central nervous system disorders, dementias, or alzheimer's disease ina mammal, said substance or composition comprising 5e-pregnane-3f,20R-diol 20-O-3-D-glucuronide or a pharmaceutically acceptable salt thereof, and said method comprising administering said substance or composition.

24. Use of 5a-pregnane-3[3,20S,21-triol 20-O-3-D-glucuronide or a pharmaceutically acceptable salt thereof, in the manufacture of a preparation 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.

25. Use of 5a-pregnane-3p,20R-diol 20-O-B-D-glucuronide or a pharmaceutically acceptable salt thereof, in the manufacture of a preparation 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.

26. A method of providing progestational treatment to a mammal which comprises administering a progestationally effective amount of 5o.-pregnane-3f,208S,21-triol 20-O-§- D-glucuronide or a pharmaceutically acceptable salt thereof to said mammal. 27- A method-of-providing-progestational-treatment-to-a-mammal-which-comprises administering a progestationally effective amount of 5a-pregnane-3f,20R-diol 20-O--D- glucuronide or a pharmaceutically acceptable salt thereof to said mammal.

28. A method of inhibiting cancers, central nervous system disorders, dementias, or alzheimer's disease in a mammal, which comprises administering an effective amount of 5a-pregnane-3p,20S,21-triol 20-O-B-D-glucuronide or a pharmaceutically acceptable salt thereof, to said mammal. AMENDED SHEET yp 2 PCT/US99/23467

29. A method of inhibiting cancers, central nervous system disorders, dementias, or alzheimer’s disease in a mammal, which comprises administering an effective amount of 5a-pregnane-3f3,20R-diol 20-O-3-D-glucuronide or a pharmaceutically acceptable salt thereof, to said mammal.

30. Process for the preparation of Sc-pregnane-33,(20S), 21-triol, 20-0O-6- glucuronide or a pharmaceutically acceptable thereof, which comprises reacting pregnenolone 3-acetate with lead tetraacetate in acetic acid to give the compound of formula 2: OAc Jes [o] PN 2 which is then hydrogenated over PtO; in acetic acid to give compound 3: eis o A 5 3 which is subsequently heated with excess equivalents of the compound 5: AMENDED SHEET