NL8520003A - 1-HYDROXYVITAMIN D COMPOUNDS WITH UNSATURATED SIDE CHAIN. - Google Patents

Description

5 8 5 2 0 0 0 3

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1-hydroxyvitamin D compounds with unsaturated side chain.

The invention relates to biologically active vitamin D compounds. More particularly, the invention relates to 1-hydroxyvitamin D compounds containing a 22.23-cis double bond in the side chain.

Because of the well known and clearly established activity of 1a-hydroxyvitamin D compounds in regulating the calcium and phosphate homeostasis in animals or humans, there was interest in the preparation of the natural metabolites and in finding analogues with beneficial medicinal properties . This has led to the preparation of a variety of compounds (see, for example, DeLuca et al., Topics in Current Chemistry, vol. 83, p. 1 (1979); Ann.

| 15 Rev. Biochem. 52 ', 411 (1983); Yakhimovich, Russian Chem.

i 'i I Rev. 49, 371 (1980)), of which some, for example, 1-hydroxyvitamin D 1 (US Pat. No. 3,741,996) or 1a-25-dihydroxyvitamin (US Pat. No. 3,697,559) are already used in the medical field practice. There remains interest in such Compounds exist, especially since it is now recognized that in addition to their known activity as regulators of the calcium homeostasis, 1,25-dihydroxy-vitamin D1 and its analog, 1a-hydroxyvitamin D1, also Once influence cellular differentiation processes and inhibit the growth and reproduction of certain malignant cells / Suda et al., U.S. Patent 4,391,802; Suda et al., Proc. Natl. Acad. Sci. USA 80, 201 (1983); Reitsma et al., Nature, Vol. 306, 492-494 (1983) /.

30 There is growing evidence to support the assumption that the exercise of biological activity by 8520003-2 vitamin D metabolites and analogues occurs through binding to an intracellular receptor protein at some stage of the overall process (see DeLuca et al., literature mentioned above). Thus, a high affinity for this receptor-5 protein is a prerequisite for high activity and desirable vitamin D analogs are those that compete efficiently with the natural hormone, 1.25- (011) ^ 0 ^ / for the receptor binding site. .

Known side-chain unsaturated vitamin D compounds include the hydroxy derivatives of vitamin, namely 25-hydroxyvitamin D2 (U.S. Patent 3,585,221), 1,25-dihydroxyvitamin (U.S. Patent 3,880,894), 24-hydroxy and 24.25-dihydroxyvitamin (Jones et al., Arch. Biochem.

15 Biophys. 202, 450 (1980)), 1α-hydroxyvitamin D 4 (U.S. Patent 3,907,843) and certain 24-desmethyl vitamin ID 4 compounds (U.S. Patent 3,786,062; Bogoslovskii et al., J. Gen. Chem. USSR 48 (4), 828 (1978);

Chem. Abstr. 89, 163848j and 89, 209016s). An example 11 of a compound having a cis-double bond in the side chains is also known (Bogoslovskii et al., Literature above).

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The new compounds according to the present invention are characterized by the structural formulas A and: B, indicated on the formula sheet. wherein the hydroxy-1: I group (or protected hydroxy group) on the 1-carbon atom may have the α or β-stereochemical orientation, and wherein a hydrogen or a hydroxy-protecting group, for example acyl, alkylsilyl, methoxymethyl or Tetrahydropyranyl.

These connections are thus characterized by a 22.23-cis double bond (22Z double bond) in the side chain.

. Preferred hydroxy protecting groups are acyl (alkanoyl) groups of 1-6 carbon atoms (e.g. formyl, acetyl, propionyl, butyryl, etc.) or 8520003-3-aroyl groups, such as benzoyl, halogen or nitrobenzoyl, or carboxyalkanoyl groups of 2-6 carbon atoms, such as oxalyl, malonyl, succinyl, glutaryl or adipyl.

Particularly preferred are the compounds of the above type A with a 1α-hydroxy group, because these compounds exhibit a surprisingly high affinity for the receptor proteins. These compounds, as 1-hydroxyvitamin D analogs, are related to the known 1-hydroxylated vitamin D compounds, but due to the presence of a 22Z double bond, they were expected to show low or no affinity for the receptor at all, because this 22.23-cis double bond forces the side chain into a completely different geometry than that assumed by the fully saturated side chain, as it occurs in 1-hydroxyvitamin or in the natural hormone, 1..25- (OH) 2D, both compounds with a known high affinity for the receptor proteins (DëLuca et al, literature above). Surprisingly and unexpectedly, however, it was found that the 1a-hydroxy-22Z-dehydro compounds actually have a higher affinity for the receptor than 1a-hydroxyvitamin ·

The synthesis of the new compounds of this invention is shown in Reaction Scheme A.

In the following description of this method and in the examples, the designations of the compounds by numbers (for example (1), (2), :( 3) ...., etc.) refer to the thus in reaction scheme A or numbered structures in the description.

The starting material for the synthetic process is the diene-protected aldehyde of the formula (1), wherein R is a methoxymethyl group. This starting material is prepared from ergosterol by the method of Morris et al. (J. Org. Chem. 46, 3422 (1981)). The reaction of compound (1) with a Wittig reagent of the formula (CH. ^) 2CHCH2CH2-PPh3Br 8520003-4 - in an organic solvent and in the presence of a strong base provides product (2) that provides the desired 22Z-olefinic side chain.

By removing the hydroxy protecting group under acidic conditions, the product (2) is converted to compound (3), which is subjected to a reduction with a strong hydride reducing agent in an organic solvent, the 5.7-diene sterol , compound (4), is obtained.

Irradiation of this material, dissolved in an organic solvent, with ultraviolet light converts the 5,7-diene into the corresponding previtamin intermediate, which after isolation and purification to form the 22Z-dehydro-vitamin D analog with the formula (.5) (R = H) 15 is isomerized by gentle heating in an organic solvent at a temperature ranging from room temperature to reflux temperature.

The intermediate of formula (5) is a known vitamin D analog, previously described by Bogoslovskii et al. (J. Gen. Chem. USSR, 48 (4), 828 (1978)) | has been prepared by a less suitable procedure.

Intermediate (5) is then converted to the desired final products by 1-hydroxylation using the general method of DéLuca et al.

i. ·! 2 5 (Ame o r se oc troo i schr if ten 4.195.027 and. 4.260.-549-).

| Connection (5). is first isosylated to form the 3 $ tosylate of the formula (6) which is then buffered methanol is sololysed. where the | new 3.5-eyclovitamin D intermediate of the formula (7) (R = H) is obtained. This product is then treated with selenium dioxide and tert.-butyl hydroperoxide in an organic solvent, the main product of which is the 1-hydroxycyclovitamin D analog of formula (8), wherein R is a hydroxy group. It is noteworthy that this allylic hydroxylation at the 1-carbon atom proceeds without complications for a linkage such as intermediate (7) with the unusual cis-double bond in the side chain with two allylic positions.

The intermediate 1-hydroxycyclovitamin D 5 product is then solvolysed in glacial acetic acid, combining a mixture of the 5.6-cis and 5.6-trans-vitamin D compounds of formulas (9) and (10) with a 3β-acetoxy group, respectively. is acquired. These acetate derivatives (9) and (10) are then separated and hydrolysed separately in a mild base to yield the desired free diol products, which are characterized by formulas 11 and 12, wherein X. represents hydrogen. .

It was found that the above-described 1-hydroxylated cyclovitamin D product, of which the 1a-hydroxy-3,5-cyclovitamin D compound of the formula; (8) the major component is also a small amount of the corresponding 13-hydroxy-3,5-cyclovitamin D | epimer, ie the product of the formula (13) indicated below. After solvolysis in glacial acetic acid, this 1β-hydroxy-epimer gives the corresponding 5.6-cis and 5.6-trans-18-hydroxy-3β-acetoxy-vitamin d analogs, represented by formulas (14) and (15), respectively (see reaction scheme B), which may be isolated from the solvolysis mixture, if desired, by chromatography; and then separately hydrolyzed in a mild base, as described above, to the 13.38-diol epimer, which are respectively characterized by. formulas (16) and (17).

In practice it has been found that the above-mentioned 5.6-trans-1-hydroxy-derivative of the formula (15) is often such an important component of the sololysis mixture that the direct isolation thereof can be too laborious. In such cases it is generally more suitable to use the 5.6-trans-18-rhydroxy analogs according to the known iodine-catalyzed isomerization process of Verloop et al. (Ree. Trav. Chim. Pays-Bas 78, 1004 (1969)) from the corresponding 5.6-cis compounds. Thus, the treatment of product (14) with a catalytic amount of iodine in a hydrocarbon or ether solvent gives the 5.6 trans product (15) and the analog isomerization of (16) provides the corresponding trans compound with the formula (17).

Acylated derivatives of the products of this invention can be easily prepared by conventional methods. Thus monoacylates of formulas (9), (10) or (14) and (15) result directly from solvolysis; such monoacylates can be further acylated to the corresponding 1,3r diacylates or desired acylates can be prepared by conventional acylation of the free diols of formulas (11), (12) or (16) and (17). It should also be noted that the 1-hydroxycyclovitamin D intermediates of formula (8) or (13) can be acylated to the corresponding 1-O-acyl derivatives. The subsequent solvolysis of such aeyl derivatives in glacial acetic acid or in i. An acidic aqueous medium (for example, by the method i of DëLuca et al, U.S. Patent 4,195,027) I gives the 5.6-cis and 5.6-trans-1-hydroxyvitamin D analogs: I as their 1,3-di-O, respectively -acyl or 1-0-acyl-deriva- | 25 ten.

| A remarkable property of the new compounds of this invention is their high activity

! I

power as reflected by the high binding affinity; ! protein receptor. It was postulated that the change in side chain geometry caused by the presence of a 22.23-cis double bond i (22Z double bond) would destroy the binding affinity: or at least result in a marked decrease. of the binding affinity, because it is known (see, for example, DéLuca et al., Topics in Curr. Chem., cited literature) that even subtle changes in the 8520003-7 stereochemistry (e.g., the change of 24R-hydroxy in 24S-hydroxy) can result in very large differences in binding properties, and the compounds were in fact prepared for the purpose of confirming this assumption. Surprisingly, competitive binding assays (performed according to the protocol of Shepard et al., Biochem. J. 182, 55 (1979)) found that the 1a-hydroxy-22Z-dehydro analog of formula (11) was three to five times higher affinity for the receptor protein then shows α-hydroxyvitamin D 1, a well-known and potent vitamin derivative. The other products of this invention exhibit a lower yet still significant binding affinity, which in each case is higher than that of the corresponding compound, which contains a saturated side chain as it occurs in natural metabolites or other known analogues.

Because of this high binding affinity, the compounds of this invention can be highly useful substitutes for the known metabolites; in the therapy or prophylaxis of calcium disorders, such as English disease (rickets), hypoparathyroidism, osteodystrophy, osteomalacia or osteoporosis in man, or related calcium deficiency diseases (eg milk fever) in animals. Similarly, these compounds can be used to treat certain malignancies, such as human leukemia. For the above; Applications are particularly preferred for the analog indicated in the reaction scheme A by the formula (11) or the corresponding 5.6-trans compound of the formula (12) or its acylated derivatives. Suitable mixtures of the above products can also be used in medical or veterinary applications, for example, the combination of the products represented by formulas (11) and (12).

For therapeutic purposes, the compounds may be administered by any conventional route of administration and in any form suitable for the method of administration chosen. The compounds can be formulated with any acceptable and harmless pharmaceutical carrier, in the form of pills, tablets, gelatin capsules or suppositories, or as solutions, emulsions, dispersions or suspensions in harmless solvents or oils, and the like recipes ( preparations) may also contain other therapeutic ac-10. contain beneficial and beneficial ingredients that may be suitable for the specific applications. When applied to humans, the compounds are advantageously administered in amounts of from about 0.5 to about 10 µg per day, the specific dosage being determined depending on the specific dose administered. connection, the te. treat disease and medical history, the condition and response of the patient, as will be apparent to those skilled in the art.

| The present invention is further described. 20 in the detailed description below, which is intended to be illustrative only and does not limit the scope of the invention. In this description, the physicochemical data were obtained using the methods mentioned and | 25 devices. High pressure liquid chromatography (HPLC) i

| was performed on a Waters. Associates Model ALC / GPC

204 using Zorbax-Sil (DuPont) (6.2 mm x 25 cm column, flow rate 4 ml / minute, 105 kg / cm 2). The column chromatography was performed on Silica Gel | 60, 70-230 mesh ASTM (Merck). The preparative thin layer chromatography (TLC). was performed on Silica 60 PF-254 (20 x 20 cm plates, 1 mm silica gel). The irradiations were performed using a Hanovia 608A36 mercury arc lamp equipped with a Vycor fliter. All .35 reactions are preferably conducted in an inert atmosphere. (e.g. argon).

8520003 - 9 - (22Z) -3β- (methoxymethoxy) -5α.8α- (4-phenyl-1,2-urazolo) cholesta - 6.22-diene (2).

1 sopentylphosphoniurabromide / "(CH3) 2CHCH2CH2PPh3Br_7 (1.67 g, 4.04 mmol) in dry tetrahydrofuran (73 mL) was treated with n-butyl lithium (1.7 M. solution in hexane, 2.42 mL, 4.11 mmol) at 3-5 ° C with stirring After stirring for 1 hour at room temperature, the orange-red solution was cooled to 3 ° C and aldehyde (1) (1.84 g, 3.36 mmol) in dry THF 10 (24 ml) was added The colorless reaction mixture was stirred at room temperature overnight, then poured into water and extracted with benzene.

The organic extract was washed with 5% HCl, saturated sodium hydrogen carbonate and water, dried (Na2S <Dj) and concentrated in vacuo to an oil which | was purified on a column of silica gel. Elution with a benzene-ether (94: 6) mixture provided adduct (2) (1.38 g, 68%) as a foam: NMR δ 0.83 (3H, s, 18-H3), 0.89 and 0 .91 (6H, each d, J = 6.8 Hz, 26-H3 and 27-H3), 0.97 20 (3H, d, J = 6.8 Hz, 21-Hj), 0.98 (3H , s, 19-Hj), 3.30 (1H, dd, J ^ = 4.4 Hz, J2 = 14 Hz, 9-JH), 3.38 (3H, s, OCH ^), 4.33 ( 1H, m, 3-43), 4.70 and 4.81 (2H, ABg, J = 6.8 Hz, 0CH20), 5.21 (2H, br m, 22-H and 23-H), 6 , 23 and 6.39 (2H, ABq, J = 8.5 Hz, 6-H and 7-H), 7.41 (5H, br m, Ar-H); IR: 1756; -1 + 25 1703.1601, 1397, 1046 cm; mass spectrum, m / z 601 (M, <1%), 426 (4), 364 (61), 349 (16), 253 (18), 251 (18), 119 (PhNCO, 100).

(22Z) -5α.8α- (4-phenyl-1,2-urazolo) cholesta-. 6.22-dien-33-ol (3).

30 A solution of adduct (2) (601 mg, 1 mmol) and p-toluene sulfulfonic acid (523 mg, 2.75 mmol). in a methanol (20 ml) -THF (12 ml) mixture was stirred at room temperature for 2 days. The reaction mixture was poured into saturated sodium hydrogen carbonate and extracted several times with benzene. The extracts were washed with water, dried (Na2SO4) and evaporated under reduced pressure. The purification of the crude product by column chromatography (benzene ether 70:30 as eluent) gave the hydroxy adduct (3) (550 mg, 99%) as a foam: NMR δ 0.83 (3H, s, 18 -H ^), 0.89 and 0.91 (6H, each d, J = 6.8 Hz, 26-H3 and 27-H3), 0.95 (3H, s, 19-H. ^), 0.98 (3H, d, J = 6.8 Hz, 21-H3), 3.16 (1H, dd, 0 ^ = 4.4 Hz, J2 = 14 Hz, 9-H), 4.44 ( 1H, m, 3-H), 5.22 (2H, br m, 22-H and 23-H), 6.22 and 6.39 (2H, ABq, J = 8.5 Hz, 6-H and 7-H), 7.40 -1 (5H, br m, Ar-H); IR: 3447, 1754, 1700, 1600, 1397 cm; 10 mass spectrum, m / z (557 (M +, <1%), 382 (35), 349 (33), 253 (20), 251 (33), 119 (100), 55 (82).

(22Z) -cholesta-5.7.22-trien-30-ol (4).

The adduct (3) (530 mg, 0.95 mmol) was converted to the diene (4) by reduction with lithium aluminum hydride (1 g) in tetrahydrofuran (60 ml) under reflux for 18 hours. After the usual work-up I, the product was purified by chromatography on silica (benzene-ether 94: 6 as eluent.), wherein pure diene is (4) (290 mg, 76%) was obtained after crystallization from ethanol: mp = 148-151 ° C; I / a = -132 ° (c = 0.9, CHCl 3); NMR δ 0.66 (3H, s, 18-H3): 0.90 and 0.91 (6H, each d, J = 6.8 Hz, 26-H3 and 27-H3), 0.96 (3h, s, 19-H3), 0.98 (3H, d, J = 6.9 Hz, 21-Hj), 3.64 (1H, m, 3-H), | 5.20 (2H, br m, 22-H and 23-H), 5.39 and 5.57 (2H, ABq, J = 6 I 25 Hz, 7-H and 6-H); UV λ 281 nm; IR: 3346, 1463, 1375, max max. + 1364, 1067, 1040, 831 cm; mass spectrum, m / z 382 (M, | 100), 349 (65); 323 (32), 271 (15), 253 (30).

(5Z, 7E, 22Z) -9.10-secocholesta-5.7.10 (19) .22-: tetraen-33-ol (5). ~ 30 Irradiation of 5.7-diene (4) (150mg, 0.39mmol) dissolved in ether (120ml) and benzene (30ml) (degassed with argon for 40 minutes) was performed at 0 ° C for 13 minutes using a UV lamp and a Vycor filter. HPLC (1% 2-propanol in hexane) of the resulting mixture gave the previtamin (56.9 mg, 38%) as a colorless oil: NMR δ 0.75 (3H, s, 18-CH3), 52520003 - 11 - 0.90 and 0.91 (6h, each d, J = 6.7 Hz, 26-H3 and 21-Kj, 0.99 (3H, d, J = 6.8 Hz, 21-H3), 1.64 (3H, s, 19-H3), 3.90 (1H, m, 3-H), 5.20 (2H, br m, 22-H and 23-H), 5.69 and 5.95 ( 2H, ABq, J = 12 Hz, 7-H and 6-H); UV λ 261 nm, ^ max 5 λ, 234 nm.

min

The thermal isomerization of this previtamin intermediate (56 mg, 0.15 mmol) in refluxing ethanol (3 hours) gave the oily vitamin analog (5) (43 mg, 77%) after separation by 10 HPLC. NMR δ 0.60 (3H, s, 18-H3), 0.89 and 0.90. (6H, each d, J = 6.7 Hz, 26-H3 and 27-H3), 0.97 (3H, d, J = 6.6 Hz, 2HH):,.

3.96 (1H, s, 3-H), 4.82 and 5 * 05 (2H, each narrow m, 19- ^), 5.20 (2H, br m, 22-H and 23-H), 6, 04 and 6.24 (2H, ABq, 1 J = 11.4 Hz, 7-H and 6-H); UV λ 265.5 nm, λ. 228 nm; IR: max. Mm 15 3427, 1458, 1379, 1048, 966, 943, 892 cm; mass spectrum,; m / z 382 (M +, 21), 349 (5), 271 (8), 253 (14), 136 (100),; 118 (82).

1-hydroxylation of compound .. (. 5).,

Freshly recrystallized p-toluenesulfonyl-20 chloride (50mg, 0.26mmol) was added to a solution of vitamin (5) (50mg, 0.13mmol) in dry pyridine: (300µl). After 30 hours at 4 ° C, the reaction mixture was stirred; stirring poured on ice / saturated NaHCO3. The mixture was stirred for 15 minutes and extracted with benzene. The organic extract was washed with saturated NaHCO 3, saturated copper sulfate and water, dried (Na 3 SO 4) and concentrated in vacuo to give the oily tosylate (6). The crude tosylate (6) was treated with NaHCC> 3 (150mg) in anhydrous methanol (10ml) and the mixture was stirred at 55 ° C for 8.5 hours. After cooling and concentration to about 2 ml, the mixture was diluted with benzene (80 ml), washed with water, dried (Na 2 SO 4) and evaporated under reduced pressure. The oily 3.5-cyclo-35 thus obtained. vitamin D compound (7) was sufficiently pure to be able to be used in the next oxidation step without any purification

852000J

- 12 - used. To a vigorously stirred suspension of SeO 2 (5.1 mg, 0.046 mmole) in dry C 1 Cl 2 (5 ml) was added tert-butyl hydroperoxide (16.5 µl, 0.118 mmole). After 30 minutes, dry pyridine (50 µl) was added and the mixture was stirred at room temperature for an additional 25 minutes, diluted with CH 2 Cl 2 (3 ml) and cooled to 0 ° C.

The crude 3.5-cyclovitamin product (7) in CH 2 Cl 2 (4.5 ml) was then added. The reaction was allowed to proceed at 0 ° C for 15 minutes and then the temperature of the reaction mixture was allowed to slowly rise (30 minutes) to room temperature. The mixture was transferred to a separatory funnel and shaken with 30 ml of 10% NaOH.

Ether (150 ml) was added and the separate organic phase was washed with 10% NaOH and water and dried over Na 2 SO 4. Evaporation to dryness in vacuo gave a yellow oily residue which was purified on silica gel TLC plate developed in hexane-ethyl acetate (7: 3) to give the 1-hydroxycyclovitamine product (20 mg, 37%): NMR δ 0.59 (3H, s, 18-Hj), 20 0 .63 (1H, m, 3-H), 0.89 and 0.90 (6H, each d, J = 6.9, I .... '! 26-H3 and 27' H3) '0, 96 (3H, d, J = 6.9 Hz, 21-Hj), 3.25 (3H, s,! -OCH), 4.17 (2H, m, 1-H and 6-H), 4 .96 (1H, d, J = 9.3 Hz; 7-H), 5.1-5.4 (4H, br m, 19-H2, 22-H and 23-H); mass spectrum, m / z 412 (M +, 26), 380 (48), 339 (22), 269 (28), 125 (20), 135 (100). This product consists essentially of the α-hydroxycyclovitamin D compound of the formula (8), as well as a small amount of the corresponding β-hydroxy epimer (13). These components can be separated in this step if desired, but no such separation is necessary.

The 1-hydroxycyclovitamin product (18 mg) as obtained above was heated (55 ° C / 15 minutes) in glacial acetic acid (0.8 ml), the mixture was neutralized (ice / saturated NaHCO2) and extracted with benzene and ether, where after an HPLC (1.5% 2-propanol in hexane as eluent). separation of the pure 3β-acetoxyvitamins (9) 8520003 Η - 13 - Ί “(6.60 mg, 34%, elution at 42 ml), (10) (4.20 mg, 22%, elution at 50 ml) and ( 14) (1.44 mg, 7%, elution at 36 ml) were obtained.

Compound (9): NMR δ 0.60 (3h, s, 18-H ^), 0.90 and 0.92 (6H, each d, J = 7.0 Hz, 26-H3 and 27-H) , 0.97 (3H, d, J = 6.8 Hz, 21-H3), 2.04 (3H, s, -OCOCH3), 4.41 (1H, m, 1-H), 5.02 ( 1H, narrow m, 19-H), 5.1-5.4 (4H, br m, 3-, 19-, 22- and 23-H), 6.03 and 6.35 (2H, ABq, J = 1.14 Hz, 7-H) and 6-H); UV λ 264.5 nm, λ. 227.5 nm; mass spectrum max mm 10 m / z 440 (M, 10), 380 (72), 362 (7), 269 (31), 251 (12), 135 (100), 134 (99).

Compound (10): NMR δ 0.60 (3h, s, 18-H3), 0.90 and 0.91 (6Η, each d, J = 7.0. Hz, 26-H3 and 27-H3), 0.97 (3H, d, J = 6.9 Hz, 21-H3), 2.05 (3H, s, -OCOCH ^, 4.49 (1H, m, 1-H), 5.00 and 5.14 (2H, each narrow m, 19- ^), 5.20 I (3H, br m, 3-, 22- and 23-H), 5.82 and 6.59 (2H, ABq, J = 12.0 µ Hz, 7-H and 6-H); UV λ y 228 nm; mass spectrum, m / z 440 (M +, 4), 380 (30), 269 (10), 135 (100), 134 ( 52).

Compound (14): NMR δ 0.58 (3H, s, 18-H3), 0.89 and 0.90 (6H, each d, J = 6.9 Hz, 26-H3 and 27-H3), 0.96 (3H, d, J = 6.9 Hz, 21-H3), 2.06 (3H, s, -OCOCH ^), 4.16 (1H, m, 1-H), 4.98 ( 2H, m, 3-H and 19-H), 5.1-5.4 (3h, br m, 19-, 22- and 23-H); UV λ 263 nm, λ. 227 nm; mass spec-: max min trum, m / z 440 (M, 32), 380 (78), 362 (21), 269 (28), 25 251 (19), 135 (100), 134 (82).

Hydrolysis of the 3β-acetoxy group in compound; things (9),. (10) and (14).

Each of the 33-acetoxy derivatives (9), (10) and i (14) was separately hydrolyzed using the same procedure. A solution of 38-acetoxyvitamin (0.7-6 ngX in ethanol (0.1 ml) was treated with 10% KOH in methanol (0.8 ml) and the mixture was heated at 50 ° C for 1 hour. After the usual work-up and the final HPLC purification (8% 2-propanol in hexane 35 as an eluent), the corresponding 1-hydroxy vitamins were obtained, namely: 8520003-1-4-1.

Compound (11): NMR δ 0.59 (3H, s, 18-H3), 0.89 and 0.90 (6H, each d, J = 7.0 Hz, 26-H and 27-H), 0 .96 (3H, O * 2 d, J = 6.8 Hz, 21-H3), 4.23 (1H, m, 3-H), 4.43 (1H, m, 1-H), 5, 00 (1H, narrow m, 19tH), 5.1-5.4 (3H, br m, 19-, 22-, and 5 23-H), 6.02 and 6.39 (2Η, ABq, J = 11.4 Hz, 7-Η and 6-H); UV λ 264.5 nm, λ. 227.5 nm; mass spectrum, m / z 398 ^ max min (M, 21), 380 (8), 287 (6), 269 (7), 251 (5), 152 (36), 134 (100). (Elution volume 39 ml.

Compound (12): NMR δ 0.61 (3H, s, 18-H3), 0.89 and 0.91 (6H, each d, J = 7.0 Hz, 26-H3 and 27-H ^) , 0.97 (3H, d, J = 6.9 Hz, 21-H3), 4.25 (1H, m, 3-H), 4.51 (1H, m, 1-H), 4.98 and 6.13 (2H, each narrow m, 19-H2), 5.21 (2H, br m, 22-H and 23-H), 5.89 and 6.59 (2H, ABq, J = 11, 5 Hz, 7-H and: 6-H); UV λ 273 nm, λ, 229.5 nm; mass spectrum, m / z max min 15 398 (M, 17), 380 (4), 287 (5), 269 (5), 251 (4), 152 (29), | 134 (100). (Elution volume 38 ml).

: Compound (16): NMR δ 0.60 (3H, s, 18-H3), 0.89 and 0.91 (6H, each d, J = 7.0 Hz, 26-H3 and 27-H3), 0.97 (3H, d, J = 6.9 Hz, 21-H3), 4.10 (1H, m, 3-H), 4.36 (1H, m, 1-H), 5.01 (1H, d, J = 2 Hz, 19-H), 5.1-5.4 (3H, br m,

19-, 22- and 23-H), 6.06 and 6.45 (2H, ABq, J = 11.3 Hz, 7-H

and 6-H); UV λ 262.5 nm, λ, 226.5 nm; mass spectrum, max min m / z 398 (M, 20), 380 (19), 269 (11), 251 (10), 152 (100), 34 (60). (Elution volume 32 ml).

If desired, the compounds of this invention can be easily obtained by crystallization from suitable solvents, such as ethers, hexane, alcohols and mixtures thereof, as will be apparent to those skilled in the art.

30 8520003

Claims (13)

1. J 1. Compounds of the formula A or B, j in which and are hydrogen or acyl and in which the substituent on carbon-1 may have the stereochemical orientation. I | 5 2. Compounds according to claim 1, wherein j are X and hydrogen. Compounds according to claim 1, wherein at least one of X 1 and X 1 is acetyl. j 4. A compound of the formula C, wherein X ^ and | A compound according to claim 4, wherein X 1 and X 1 are hydrogen. I A compound according to claim 5 in crystalline form. 7. Compounds of the formula D, wherein I Xj and X ^ are hydrogen or acyl. A compound according to claim 6, wherein X, and X_ are hydrogen. A pharmaceutical composition comprising at least one of the compounds of claim 4 in addition to a pharmaceutically acceptable excipient. A pharmaceutical composition comprising the compound of claim 5 in addition to a pharmaceutically acceptable excipient. X2 are hydrogen or acyl. j A pharmaceutical composition, comprising at least one of the compounds of claim 7 in addition to a pharmaceutical acceptable excipient. 12. A pharmaceutical composition comprising the compound of claim 8 in addition to a pharmaceutically acceptable excipient. A pharmaceutical composition comprising a mixture of the compounds of claim 5 and claim 8 and a pharmaceutically acceptable excipient. ; 8520003