JPH0455424B2 - - Google Patents

Description

Claim 1: Compound having the following formula:

[Formula] or

[Formula] (In the formula, X ₁ and X ₂ are hydrogen or acyl, and the substituent on carbon 1 has an α or β stereochemical configuration.) 2 A patent claim in which X ₁ and X ₂ are hydrogen Range 1
Compounds described in Section. 3. The compound according to claim 1, wherein at least one of X ₁ and X ₂ is acetyl. 4. A compound according to claim 1 having the following formula: (In the formula, X ₁ and X ₂ are hydrogen or acyl.) 5. Claim 4 where X ₁ and X ₂ are hydrogen.
Compounds described in Section. 6. The compound according to claim 5, which is in a crystalline form. 7. A compound according to claim 1 having the following formula. (In the formula, X ₁ and X ₂ are hydrogen or acyl.) 8. Claim 6 in which X ₁ and X ₂ are hydrogen.
Compounds described in Section. This invention was made possible by the NIH Award from the Department of Health and Human Services.
This work was made with Government support under Grant No. AM14881. The government has certain rights in this invention. TECHNICAL FIELD This invention relates to biologically active vitamin D compounds. More specifically, the side chain contains 22,23-cis-
1-Hydroxyvitamin D compounds containing double bonds. BACKGROUND Because of the well-known and well-documented activity of the 1α-hydroxyvitamin D compound in the regulation of calcium and phosphate homeostasis in animals and humans, the preparation of its natural metabolites and analogues with useful medical properties are essential. There has been interest in the discovery of This has led to the preparation of a variety of compounds (e.g. DeLuca et al., Topics in Current
Chemistry, Vol. 83 , p1 (1979); Ann.Rev.
Biochem. 52 , 411 (1983); Jakimovich,
Russin Chem. Rev. 49 , 371 (1980). Among them, for example 1α-hydroxyvitamin D ₃ (US Pat. No. 3,741,996) or 1α,25-dihydroxyvitamin D ₃ (US Pat. No. 3,697,559) have already found use in their medical practice. Of particular interest in these compounds is 1α,25-dihydroxyvitamin _D3 and its analogues, 1α-hydroxyvitamin _D3 , in addition to their classical function as regulators of calcium homeostasis.
It has been and continues to be found that cell membranes can also affect the cell identification process and inhibit the growth and proliferation of certain malignant cells (Suda et al., US Pat. No. 4,391,802). Sudha et al. Proc.
Natl.Acad.Sei.USA 80 , 201 (1983); Reitsma et al.
Nature, Vol. 306 , p. 492-494 (1983)). Increasing evidence indicates that vitamin D metabolites and their analogues exhibit biological activity through binding to intracellular receptor proteins at some stage in the overall process. (DeLuca et al., supra). A high affinity for this receptor protein is thus a prerequisite for high efficacy, and desirable vitamin D analogs have a high affinity for receptor binding sites, such as natural hormones,
It competes effectively with 1,25-(OH) ₂ D ₃ . Known side chain unsaturated vitamin D compounds are vitamin
Hydroxy derivatives of _D2 , i.e., 25-hydroxyvitamin _D2 (US Pat. No. 3,585,221), 1α,25-
Dihydroxyvitamin D ₂ (US Patent No. 3880894)
), 24-hydroxy- and 24,25-dihydroxyvitamin D ₂ (Gyons et al., Arch.Biochem.
Biophys 202 , 450 (1980)), 1α-hydroxyvitamin D ₂ (U.S. Pat. No. 3,907,843) and certain
24-Demethylated Vitamin D ₂ Compound (U.S. Patent No.
3786062; Bogoslovski et al., J.Gen.Chem.
USSR 48 (4), 828 (1978); Chem Abstr. 89 ,
163848j and 89 , 209016s). cis -2 in side chain
An example of a compound with a double bond is also known (Bogoslovski et al., supra). DISCLOSURE OF THE INVENTION The novel compounds of the present invention are characterized by structures A and B shown below. (wherein the hydroxy group (or protected hydroxy group) at carbon 1 may have an α- or β-stereochemical configuration, and X ₁ and X ₂ are hydrogen or hydroxy protecting groups, e.g. alkylsilyl ,
Indicates methoxymethyl or tetrahydropyranyl group. ) These compounds thus
It is characterized by a 22,23-cis-double bond (22Z-double bond). Preferred hydroxy protecting groups include acyl (alkanoyl) groups having 1 to 6 carbon atoms (e.g. formyl,
acetyl, propionyl, butyryl, etc.) or an aroyl group such as benzoyl, halo or nitrobenzoyl, or a carboxyalkanoyl group having 2 to 6 carbon atoms such as oxalyl, malonyl, succinyl, glutaryl or adipyl. Particularly preferred are compounds of type A, as described above, having a 1α-hydroxy group. This is because these compounds have an unexpectedly high affinity for receptor proteins.
These compounds are related to 1-hydroxylated vitamin D compounds known as 1-hydroxyvitamin D analogs. However, due to the presence of the 22Z-double bond, the 22,23-cis-double bond binds the side chain to the 1α-hydroxyvitamin, which is known to have a high affinity for receptor proteins (DeLuca et al.).
D ₃ or the natural hormone 1,25-(OH) ₂ Because the side chains of both compounds, such as _the fully saturated side chains, result in a completely different geometry than would be considered, it is possible that the affinity for the receptor is However, it was expected to be low. However, 1α
It has been found that the -hydroxy-22Z-dehydro compound actually exhibits a higher affinity for its receptor than does 1α-hydroxyvitamin _D3 . The synthesis of the novel compounds of the invention is outlined in the process scheme. The following description of this process and the numbers in each example (e.g. (1), (2),
(3) etc.)
Refers to the scheme or structure so numbered in the specification. The starting material for this synthetic method is structure (1), where R
is a diene-protected aldehyde with a methoxymethyl group). This starting material was prepared using the method of Morris et al. (J.
Prepared from ergosterol by Org.Chem. 46 , 3422 (1981). When compound (1) is reacted with the Wittig reagent having the structure shown below (CH ₃ ) ₂ CHCH ₂ CH ₂ PPh ₃ Br in an organic solvent in the presence of a strong base, the desired reaction is achieved.
22Z-Products characterized by olefin side chains (2)
give. Removal of the hydroxy protecting group under acidic conditions converts the product (2) to compound (3), which is then subjected to reduction with a strong hydride reducing agent in an organic solvent5.
7-Diesterol, compound (4) is obtained. When this substance is dissolved in an organic solvent and irradiated with ultraviolet light, the 5,7-diene is converted to the corresponding previtamin intermediate, which, after isolation and purification, can be produced in an organic solvent at room temperature to reflux temperature. It is isomerized by mild heating at temperatures to 22Z-dehydro-vitamin _D3 analogs of structure (5). An intermediate with structure (5), known as a vitamin D analog, was previously prepared by Bokoslovsky et al. (J. Ghen. USSR, 48 (4), 828
(1978)), which was a rather inconvenient method. Intermediate (5) is then converted to the desired final product by 1-hydroxylation using the general method of DeLuca et al. (US Pat. Nos. 4,195,027, 4,260,549). Compound (5) is first tosylated to give the structure
(6), which then gives the 3β-tosylated product with
Structure (7) subjected to solvolysis in buffered methanol.
A novel 3,5-cyclovitamin D intermediate (R=H) is obtained. This product was then treated with selenium dioxide and tert-butyl hydroperoxide in an organic solvent to obtain 1α-hydroxycyclovitamin D of structure (8), where R is a hydroxy group.
Obtain analogues. This allylic hydroxylation at carbon (1) causes complications for compounds with an exceptional cis-double bond in the side chain, such as intermediate (7) with two allylic positions. It is noteworthy that the process progresses without any problems. The 1-hydroxycyclovitamin D product is then subjected to solpolysis in glacial acetic acid and has a 3β-acetoxy group.
5,6-cis and 5,6-cis of structures (9) and (10), respectively.
A transvitamin D compound is obtained in a mixed state. These acetate derivatives (9) and (10) are then fractionated and Individually, the desired free diol products characterized by structures (11) and (12) (where X ₁ represents hydrogen) are produced in a mild base. Among the above 1-hydroxylated cyclovitamin D products, 1α-hydroxy-3,5- of structure (8)
It was found that cyclovitamin D was the main component and also contained a small amount of the corresponding 1β-hydroxy-3,5-cyclovitamin D epimer, a product of structure (13) below. Upon solvolysis in glacial acetic acid, this 1β-hydroxy-epimer was converted into the corresponding 5,6-cis and 5,6-trans-1β-hydroxy-3β-
Generates acetoxy-vitamin D analogs,
It can be isolated from the solvolysis mixture by chromatography, if desired, and then hydrolyzed separately in mild base as described above and characterized by structures (16) and (17), respectively. It can be made into a 1β,3β-diol epimer that can be attached. In practice, the 5,6-trans-
Since 1β-hydroxy derivatives often represent such a minor component of the solvolysis mixture,
Its direct isolation was extremely difficult. In such cases, 5,6-trans-1β-hydroxy analogues can be used as described by Verloop et al. (Rec. Trav-
More conveniently, they are prepared from the corresponding 5,6-cis compounds by the iodine-catalyzed isomerization process of Chim. Pays-Bas 78 , 1004 (1969). Thus, treatment of product (14) with a catalytic amount of iodine in a hydrocarbon or ether solvent gives the 5,6-trans product (15), and analogous isomerization of (16) gives the corresponding structure ( 17) gives the trans compound. Acylated derivatives of the products of this invention are readily prepared by conventional methods. Therefore, the structure
Monoacylation of (9), (10) or (14) and (15) occurs directly by solvolysis. Such monoacylated product can be further acylated to the corresponding 1,3-diacylate, or the desired acylated product can be further acylated to the corresponding 1,3-diacylate.
They are prepared by conventional acylation of free diols of structure (11), (12) or (16), (17). It should also be noted that the 1-hydroxycyclovitamin D intermediates of structure (8) or (13) can be acylated to the corresponding 1-o-acyl derivatives. Subsequent solvolysis of such acyl derivatives in glacial acetic acid or acidic aqueous catalysts (e.g., DeLuca et al., U.S. Pat. No. 4,195,027)
5,6-cis and 5,6-trans-1-hydroxyvitamin D analogues of their 1,3-di-o
-acyl or 1-o-acyl derivatives, respectively. A notable property of the novel compounds of the present invention is their high potency as demonstrated by their high binding affinity for protein receptors.
The change in the geometric isomerism of the side chain determined by the presence of a 22,23-cis-double bond (22Z-double bond)
It was expected that binding affinity would be lost or at least binding affinity would be significantly reduced. For it is known that even subtle changes in stereochemistry (e.g. from 24R-hydroxy to 24S-hydroxy) can produce appreciable differences in binding affinity (e.g. DeLuca et al. , Topics in Curr.Chem. above)
It is from. When these compounds were actually prepared to confirm this speculation, it was surprisingly found that the 1α-hydroxy-22Z-dehydro analogue of structure (11) was a well-known highly potent vitamin D ₃ derivative, 1α. Competitive binding analysis (conducted according to the protocol of Siephardt et al., Biochem _. J. 182 , 55
(1979)). Other products of the invention exhibit lower binding affinities, but still substantially higher than the respective binding affinities of corresponding compounds featuring saturated side chains, such as natural metabolites or other known analogues. Has binding affinity. Due to their high binding affinity, the compounds of the present invention can be used to treat calcium disorders such as rickets, hyperparathyroidism, osteodystrophy, osteomalacia, osteoporosis in humans or related calcium deficiencies (in animals). It is extremely useful as an alternative to known metabolites in the treatment or prevention of lactation fever, for example. Similarly, these compounds can be used to treat certain malignant diseases such as leukemia in humans. Suitable for the above use are structures in the process scheme.
5, 6 of the analogue drawn in (11) or the counterpart of structure (12)
-trans- compounds, or acylated derivatives thereof. Suitable mixtures of the above products can also be used for medical or veterinary applications. For example, it is a combination of compounds represented by structures (11) and (12).
For therapeutic purposes, they may be administered by any conventional route and in any form suitable for the chosen mode of administration. The compounds may be formulated with an acceptable and non-toxic pharmaceutical carrier in the form of pills, tablets, gelatin capsules, or suppositories, and also as solutions, emulsions, dispersions, or suspensions in non-toxic solvents or oils. The formulation may contain such therapeutically active and beneficial ingredients as may be deemed appropriate for the particular use. For human use, the compound is advantageously administered in amounts of about 0.5 to about 10 μg per day.
It will be apparent to those skilled in the art that the respective dosage will be adjusted depending on the particular compound being administered, the disease being treated, and the response of the patient. The invention is further illustrated by the following detailed description, which is illustrative only and is not intended to limit the scope of the appended claims. The physico-chemical data herein were determined using the methods and equipment mentioned. High pressure liquid chromatography (HPLC) is a Waters Associates model using Zorbax-Sil (DuPont).
ALC/GPC204 (6.2mm x 25cm column, flow rate 4ml/
min, 1500psi). Column chromatography is silica gel 60, 70~230 mesh
It was performed on ASTM (Merck & Co.). Separative thin layer chromatography (TLC) uses silica 60PF-254 (20
x 20 cm plate, 1 mm silica gel).
Light irradiation was performed using a Hanobia 608A36 mercury arc lamp fitted with a Vicor filter. All reactions are preferably carried out under an inert atmosphere (eg argon). (22Z)-3β-(methoxymethoxy)-5α,8α-
(4-phenyl-1,2-urazolo)cholester 6,22-diene (2) Isopentylphosphonium bromide [(CH ₃ ) ₂ CHCH ₂ CH ₂ PPh ₃ Br] (1.67 g) in dry tetrahydrofuran (73 ml) ，
4.04 mmol) was treated with n-butyllithium (1.7M in hexane, 2.42 ml, 4.11 mmol) at 3-5°C with stirring. After stirring at room temperature for 1 hour, the orange red solution was cooled to 3°C and dissolved in aldehyde (1) in dry THF (24 ml).
(1.84g, 3.36mmol) was added. The colorless reaction mixture was stirred at room temperature overnight and then poured into water.
Extracted with benzene. 5% of its organic extract
Washed with HCl, saturated sodium bicarbonate, and water, dried (Na ₂ SO ₄ ), and concentrated to an oil under reduced pressure. And it was purified on a silica gel column. Elution with a benzyl-ether (94:6) mixture gave the adduct (2). (1.38g,
68%) NMRδ0.83 (3H, s, 18−H ₃ ) as bubbles,
0.89 and 0.91 (6H, respectively d, j = 6.8Hz, 26−H ₃ and 27
−H ₃ ), 0.97 (3H, d, J = 6.8Hz, 26−H ₃ and 27−
_H3 ), 0.97 (3H, d, j=6.8Hz, 21- _H3 ), 0.98
(3H, a, 19-H ₃ ), 3.30 (1H, dd, J ₁ = 4.4Hz,
_J2 = 14Hz, 9-H), 3.38 (3H, s, _OCH3 ), 4.33
(1H, m, 3-H), 4.70 and 4.81 (2H, ABq, J=
6.8Hz, OCH ₂ O), 5, 21 (2H, brm, 22-H and
23-H), 6.23 and 6.39 (2H, ABq, J=8.5Hz, 6
-H and 7-H), 7.41 (5H, brm, Ar-H); IR:
1756, 1703, 1601, 1397, 1046 cm ^-1 ; Mass spectrum, m/z601 (M ⁺ , <1%), 4.26(4), 364(61),
349(16), 253(18), 251(18), 119 (PhNCO, 100). (22Z)-5α,8α-(4-phenyl-1,2-urazolo) cholesta-6,22-dien-3β-ol
(3) A mixture of a solution of adduct (2) (601 mg, 1 mmol) and p-toluenesulfonic acid (523 mg, 2.75 mmol) in methanol (20 ml)-THF (12 ml) was prepared at room temperature for 2 hours.
Stirred for days. The reaction mixture was poured into saturated sodium bicarbonate and extracted several times with benzene. The extracts were washed with water, dried (Na ₂ SO ₄ ) and evaporated under reduced pressure. The crude product was purified by column chromatography (benzene ether 70:30 as eluent) to give adduct (3) (550 mg, 99%) as a foam. NMRδ0.83 (3H, a, 18−H ₃ ), 0.89
0.91 (6H, each d, J = 6.8Hz, 26−H ₃ and 27−
H ₃ ), 0.95 (3H, a, 19-H ₃ ), 0.98 (3H, d, J
= 6.8Hz, 21−H ₃ ), 3.16 (1H, dd, J ₁ = 4.4Hz, J ₂
=14Hz, 9-H), 4.44 (1H, m, 3-H), 5.22
(2H, brm, 22-H and 23-H), 6.22 and 6, 39
(2H, ABq, J=8.5Hz, 6-H and 7-H), 7.40
(5H, brm, Ar-H); IR: 3447, 1754, 1700,
1600, 1397cm ^-1 ; Mass spectrum, m/z (557
(M ⁺ , <1%), 382(35), 349(33), 253(20), 251(3
3), 119(100), 55(82). (22Z)-Corestar-5,
The 7,22-trien-3β-ol (4) adduct (3) (530 mg, 0.95 mmol) was reduced with lithium aluminum hydride (1 g) in tetrahydrofuran (60 ml) under reflux for 18 hours to give the diene (4). Converted to . After the usual work-up, the product was purified by chromatography on silica gel (benzene-ether 94:6 as eluent) and crystallized from ethanol to give the pure diene (4).
(Given 290 mg, 76%. m.p148-151℃ [α] ²⁴ _D =
132℃ (c=0.9, CHCl ₃ ); NMR δ0.66 (3H, s,
18−H ₃ ), 0.90 and 0.91 (6H, each d, J = 6.8Hz), 26
−H ₃ and 27−H ₃ ), 0.96 (3H, s, 19−H ₃ ), 0.98
(3H, d, J=6.9Hz, 21−H ₃ ), 3.64 (1H, m,
3-H), 5.20 (2H, brm, 22-H and 23-H),
5.39 and 5.57 (2H, ABq, J=6Hz, 7-H and 6-
H); UVλ _nax 281nm; IR: 3346, 1463, 1375,
1364, 1067, 1040, 831cm ^-1 ; Mass spectrum m/z382 (M ₊ , 100), 349(65), 323(32), 271(15),
253(30). (5Z, 7E, 22Z) −9, 10 − Secoco Star 5,
7,1019,22-Tetraen-3β-ol 5 5,7-Diene (4) (150 mg, 0.39 mol) was dissolved in ether (120 ml) and benzene (30 ml) (degassed with argon for 40 min) at 0°C. Light was irradiated for 13 minutes using a UV-lamp and a Vicol filter. The resulting mixture was subjected to HPLC (1% 2-propanol in hexanes) to yield the previtamin (56.9 mg, 38%) as a colorless oil. NMRδ0.75 (3H, s, 18−
CH ₃ ), 0.90 and 0.91 (6H, each d, J = 6.7Hz, 26−
H ₃ and 27−H ₃ ), 0.99 (3H, d, J = 6.8Hz, 21−
H ₃ ), 1.64 (3H, s, 19−H ₃ ), 3.90 (1H, m, 3
-H), 5.20 (2H, brm, 22-H and 23-H), 5.69
and 5.95 (2H, ABq, J=12Hz, 7-H and 6-
H); UV λ _nax 261 nm, λ _nio 234 nm. This previtamin intermediate (56 mg, 0.15 mmol)
After thermal isomerization in refluxing ethanol (3 hours) and separation by HPLC, the oily vitamin analog (5) was obtained.
(43 mg, 77%). NMRδ0.60 (3H, a,
18−H ₃ ), 0.89 and 0.90 (6H, respectively d, J = 6.7Hz,
26−H ₃ and 27−H ₃ ), 0.97 (3H, d, J = 6.6Hz, 21
-H ₃ ), 3.96 (1H, s, 3-H), 4.82 and 5.05 (2H,
respectively narrow m, 19-H ₂ ), 5.20 (2H, brm, 22-H
and 23−H), 6.04 and 6.24 (2H, ABq, J=11.4Hz,
7-H and 6-H); UVλ _nax 265.5 _on λ _nio 228nm;
IR: 3427, 1458, 1379, 1048, 966, 943,
892cm ^-1 ; Mass spectrum, m/z382 (M ⁺ , 21),
349(5), 271(8), 253(14), 136(100), 118(82). 1-Hydroxylation of Compound 5 Freshly recrystallized p-toluenesulfonic acid chloride (50 mg, 0.26 mmol) was added to a solution of vitamin (5) (50 mg, 0.13 mmol) in dry pyridine (300 μl). After 30 hours at 4° C., the reaction mixture was poured onto ice/saturated _Na HCO ₃ with stirring. The mixture was stirred for 15 minutes and extracted with benzene. The organic extracts were washed with saturated Na _{HCO 3} , saturated copper sulfate and water, dried (Na ₂ SO ₄ ) and concentrated under reduced pressure to give the oily tosylate (6). This crude tosylated product
(6) in anhydrous methanol (10 ml) with N _a HCO ₃ (150 mg)
and the mixture was stirred at 55° C. for 8.5 hours. After cooling and concentration to ~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-cyclovitamin D compound thus obtained was ten parts pure and could be used in the next oxidation step without purification. To a vigorously stirred suspension of _SeO2 (5.1 mg, 0.046 mmol) in dry _CH2Cl2 (5 _ml ) was added tert-butyl hydroperoxide (16.5 [mu]l, 0.118 mmol). Dry pyridine (50 μl) was added for 30 min and the _mixture was stirred _at room temperature for a further 25 min.
The 3.5-cyclovitamin product (7) was then added.
The reaction proceeded at 0° C. for 15 minutes, then it was slowly warmed to room temperature (30 minutes). The mixture was transferred to a separatory funnel, 30 ml of 10% NaOH was added and shaken. Ether ₍ 150ml) was added and the separated organic layer was washed with 10% NaOH, water and dried over _Na2SO4 . Concentration by drying under reduced pressure gave a yellow oily residue. It was purified on a silica gel TLC plate developed with 7:3 hexane-ethyl acetate to yield the 1-hydroxycyclovitamin product (20
mg, 37%). NMRδ0.59 (3H, s, 18,
H ₃ ), 0.63 (1H, m, 3-H), 0.89 and 0.90 (6H,
d, J=6.9Hz, 26− _H3 and 27− _H3 ), 0.96 respectively
(3H, d, J=6.9Hz, 21−H ₃ ), 3.25 (3H, s,
-OCH ₃ ), 4.17 (2H, m, 1-H and 6-H),
4.96 (1H, d, J=9.3Hz, 7-H), 5.1-5.4
(4H, brm, 19-H ₂ , 22-H and 23-H); mass spectrum, m/z412 (M ⁺ , 26), 380(48), 339(2
2), 269(28), 245(20), 135(100). This product is mainly composed of structure 8 and 1α-hydroxycyclovitamin D.
compound and also contains a small amount of the corresponding 1α-hydroxy-epimer (13). These components can be separated at this stage if desired, but such separation is not necessary. The 1-hydroxycyclovitamin product (18 mg) obtained above was heated (55°C/15 min) in glacial acetic acid (0.8 ml) to neutralize the mixture (ice/saturated
After extraction with benzene and ether and HPLC separation (using 1.5% 2-propanol in hexane as eluent), the pure 3β _- acetoxy-vitamin (9) (6.60 mg, 34%, 42 ml) was obtained. (eluted), (10) (4.20mg, 22%, eluted in 50ml) and (14)
(1.44 mg, 7%, eluted in 36 ml) was obtained. Compound (9): NMR δ0.60 (3H, s, 18-H ₃ ),
0.90 and 0.92 (6H, respectively d, J = 7.0Hz, 26−H ₃ and 27
−H ₃ ), 0.97 (3H, d, J=6.8Hz, 21−H ₃ ),
2.04 (3H, s, -OCOCH _-3 ), 4,41 (1H, m,
1-H), 5.02 (1H, narrow m, 19-H), 5.1-5.4
(4H, brm, 3-, 19-, 22- and 23-H), 6.03
and 6.35 (2H, ABq, J=11.4Hz, 7-H and 6-
H); UVλ _nax 264.5nm, λ _nio 227.5nm; mass spectrum, m/z440 (M ₊ , 10), 380(72), 362(7),
269(31), 251(12), 135(100), 134(99). Compound (10): NMR δ0.60 (3H, s, 18-H ₃ ),
0.90 and 0.91 (6H, respectively d, J = 7.0Hz, 26−H ₃ and 27
−H ₃ ), 0.97 (3H, d, J=6.9Hz, 21−H ₃ ),
0.97 (3H, d, J=6.9Hz, 21−H ₃ ), 2.05 (3H,
s, _-OCOCH3 ),4,49(1H,m,1,-H),
5,00 and 5,14 (2H, each narrow m, 19-H ₂ ),
5.20 (3H, brm, 3-, 22- and 23-H), 5, 82
and 6.59 (2H, ABq, J=12.0Hz, 7-H and 6-
H); UVλ _nax 270nm; λ _nio 228nm; Mass spectrum, m/z440 (M ⁺ , 4), 380(30), 269(10), 351(10
0), 134(52), Compound (14): NMR δ0.58 (3H, s,
18−H ₃ ), 0.89 and 0.90 (6H, respectively d, J = 6.9
Hz, 26−H ₃ and 27−H ₃ ), 0.96 (3H, d, J = 6.9
Hz, 21−H ₃ ), 2.06 (3H, s, −OCOCH ₃ ), 4,
16 (1H, m, 1-H), 4.98 (2H, m, 3-H and
19-H), 5.1-5.4 (3H, brm, 19-, 22- and 23-
H); UVλ _nax 263nm, λ _nio 227nm; mass spectrum, m/z440 (M ⁺ , 32), 380(78), 362(21), 269
(28), 251(19), 135(100), 134(82). Hydrolysis of the 3β-acetoxy group in compounds (9), (10) and (14) Each of the 3β-acetoxy-derivatives (9), (10) and (14) was hydrolyzed separately using the same procedure. . A solution of 3β-amothoxyvitamin (0.7-6 mg) in ethanol (0.1 ml) was added to
Treated with 10% KOH and heated the mixture at 50° C. for 1 hour. After normal work-up and final HPLC purification (8% 2-propanol in hexane as eluent), the corresponding 1-hydroxyvitamin was obtained. That is, compound (11): NMR δ0.59 (3H, s, 18-H ₃ ),
0.89 and 0.90 (6H, respectively d, J = 7.0Hz, 26−H ₃
and 27−H ₃ ), 0.96 (3H, d, J=6.8Hz, 21−H ₂ ),
4.23 (1H, m, 3-H), 4.43 (1H, m, 1-H),
5.00 (1H, narrow m, 19-H), 5.1-5.4 (3H, brm,
19−, 22− and 23−H), 6.02 and 6.39 (2H, ABq,
J=11.4Hz, 7-H and 6-H); UVλ _nax 264.5nm,
λ _nio 227.5nm; mass spectrum, m/z398 (M ⁺ ,
21), 380(8), 287(6), 269(7), 251(5), 152(32),
134(100)． (Elution volume 39ml). Compound (12): NMR δ0.61 (3H, s, 18-H ₃ ),
0.89 and 0.91 (6H, respectively d, J = 7.0Hz, 26−H ₃
and 27−H ₃ ), 0.97 (3H, d, J=6.9Hz, 21−H ₃ ),
0.97 (3H, d, J = 6.9Hz, 21-H ₃ ), 4.25 (1H,
m, 3-H), 4.51 (1H, m, 1-H), 4.98
5.13 (2H, respectively narrow m, 19−H ₂ ), 5.21 (2H,
brm, 22-H and 23-H), 5.89 and 6.59 (2H, ABq,
J=11.5Hz, 7-H and 6-H); UVλ _nax 273nm,
λ _nio 229.5nm; mass spectrum, m/z398 (M ⁺ ,
17), 380(4), 287(5), 269(5), 251(4), 152(29),
134(100)． (Elution volume 38ml). Compound (16): NMR δ0.60 (3H, s, 18-H ₃ ),
0.89 and 0.91 (6H, respectively d, J = 7.Hz, 26−H ₃
and 27−H ₃ ), 0.97 (3H, d, J=6.9Hz, 21−H ₃ ),
4.10 (1H, m, 3-H), 4.36 (1H, m, 1-H),
5.01 (1H, d, J=2Hz, 19-H), 5.1-5.4
(3H, brm, 19-, 22- and 23-H), 6.60 and 6.45
(2H, ABq, J=11.3Hz, 7-H and 6-H);
UVλ _nax 262.5nm, λ _nio 226.5nm; mass spectrum, m/z398 (M ⁺ , 20), 380(19), 269(11), 251
(10), 152(100), 134(60). (Elution volume 32ml). If desired, the compounds of the present invention can be readily obtained by crystallization from suitable solvents such as ethers, hexane, alcohols, and mixtures thereof, as is well known and apparent to those skilled in the art.