JP2010111593A - 14-epi-19-norprevitamin d3 derivative - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new vitamin D<SB>3</SB>derivative useful as a therapeutic agent of osteoporosis. <P>SOLUTION: The 19-norprevitamin D<SB>3</SB>derivative represented by formula (1) (wherein, both of R<SP>1</SP>and R<SP>2</SP>are hydrogen, one of R<SP>1</SP>and R<SP>2</SP>is hydrogen and the other is 1-6C alkyl, or R<SP>1</SP>and R<SP>2</SP>form methylene or 1-5C alkylmethylene in combination with each other) or a pharmaceutically acceptable solvate thereof are provided. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

The present invention relates to a 14-epi-19-norprevitamin D ₃ derivative useful as a pharmaceutical or a pharmaceutically acceptable solvate thereof, and a therapeutic agent for osteoporosis using the same.

The active vitamin D ₃ derivative has an action of promoting calcium absorption in the small intestine, and has actions of regulating bone resorption and bone formation in bone, and is used as a therapeutic agent for osteoporosis. Moreover, it has a parathyroid hormone (PTH) secretion inhibitory effect and is used for the treatment of secondary hyperparathyroidism with enhanced PTH. Furthermore, in addition to these effects, immunoregulatory effects, cell proliferation suppressing effects and cell differentiation inducing effects have been found, for example, treatment of diseases such as cancer, psoriasis, rheumatoid arthritis, diabetes mellitus, hypertension, acne, eczema, dermatitis, etc. Application to drugs is under consideration.

In general, the vitamin D ₃ derivative (3) exists in an equilibrium state with the pre-form (4) as shown in the following scheme 1. Incidentally, the following scheme 1 shows the case l [alpha], 25-dihydroxyvitamin D _3, which is a typical active vitamin D _3.

  Since the abundance ratio of the pre-form in this equilibrium is generally small, the applicability of the pre-form to pharmaceutical products has not been studied in detail.

By the way, it has been reported by Okamura et al. That the 14-epi form obtained by epimerizing the 14-position of the vitamin D ₃ derivative has a pre-form equilibrium and exists relatively stably (Scheme 2 below, non-patent). Reference 1).

However, although affinity to vitamin D receptor (VDR), in vivo intestinal calcium absorption, bone calcium mobilization, etc. have been reported so far (see Non-patent Document 1 and Patent Document 1). These activities were very weak compared to the active vitamin D ₃ derivative, and the pre-form (6) could not be expected to be effective as a therapeutic agent for various diseases utilizing vitamin D-like action.

On the other hand, it is known that chemical modification of the vitamin D ₃ skeleton to the 2nd position improves various activities including improvement of affinity for the vitamin D receptor (see Non-Patent Document 2). Based on this finding, chemical modification to the 2-position of the pre-vitamin D ₃ derivative (6) revealed that the activity was improved in the same manner as the vitamin D ₃ skeleton (compound (8), 236th ACS). National Meetings, August 2008).

However, this derivative (8) does not exist as a single derivative because it has an equilibrium conversion to the vitamin D ₃ skeleton (7) as shown in Scheme 3, and is considered to have a problem in application as a pharmaceutical product.

In addition, a compound in which the 14-position is in a natural configuration (the hydrogen atom is on the opposite side to the paper surface) and R ¹ and R ² are both hydrogen atoms is known (Non-Patent Document 4). It has been reported that the biological activity of the compound is very weak (Non-Patent Documents 5 and 6). This compound is different from the compound of the present invention in that the 14-position is an epi-type stereo and that R ¹ and R ² can have a substituent.

International Publication No. 95/017177 Pamphlet Journal of Medicinal Chemistry (J. Med. Chem.), 37, 2387-2393, 1994 Vitamin D Analogs in Cancer Prevention and Therapy Research in Cancer Research, 164, 289-317 2003 Nature Reviews Drug Discovery, 3, 27-41, 2004 Tetrahedron Lett., 33, 5445-5448, 1992 Journal of Bone and Mineral Research (J. Bone Miner. Res.), 8, 1009-1010, 1993 Journal of Biological Chemistry (J. Biol. Chem.), 278, 35476-35482, 2003

An object of the present invention is to provide a previtamin D ₃ derivative having a vitamin D-like action. Another object of the present invention is to provide a therapeutic agent for osteoporosis. Another object of the present invention is to provide a production intermediate of previtamin D ₃ derivatives having vitamin D-like action.

As a result of diligent research for the above purpose, the present inventors have reached the following invention.
That is, the present invention is a 19-norprevitamin D ₃ derivative represented by the following formula (1) or a pharmaceutically acceptable solvate thereof.

Here, R ¹ and R ² in the formula (1) are both hydrogen atoms, one is a hydrogen atom and the other is a C ₁ -C ₆ alkyl group, both are methylene groups, or both are together I represent a _C 1 -C ₅ alkyl methylene group.

Further, the present invention is treatment of osteoporosis containing 19-nor previtamin D ₃ derivative or a pharmaceutically acceptable solvate thereof represented by the above formula (1) as an active ingredient.

Further, the present invention is a production intermediate of the 19-nor previtamin D ₃ derivative represented by the following formula (2).

Here, R ³ and R ⁴ in the formula (2) are both hydrogen atoms, one is a hydrogen atom and the other is a C ₁ -C ₆ alkyl group, both are methylene groups, or both are together I represent a _C 1 -C ₅ alkyl methylene group.

The compounds of the present invention, the known 19-natural previtamin D ₃ derivative (6), has a strong transcriptional activity in osteoblasts as compared to (8). Further, the known 19-natural previtamin D ₃ derivative (6), there is no equilibrium conversion between vitamin D ₃ skeleton present in (8), present as a single compound. Therefore, the compound of the present invention is highly useful as a therapeutic agent for osteoporosis.

The definitions of terms in the present invention are as follows.
The alkyl group refers to a linear, branched or cyclic aliphatic hydrocarbon group. The C ₁ -C ₆ alkyl group means an alkyl group having 1 to 6 carbon atoms, such as a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a pentyl group, an isopentyl group, and a hexyl group. , Cyclopropyl group, cyclopropylmethyl group and cyclohexyl group can be mentioned as specific groups.

In the case of the present invention, a methylene group is ^one in which R ¹ and R ² are expressed together, and specifically, is a derivative as described below.

In the case of the present invention, an alkylmethylene group is ^one in which R ¹ and R ² are both represented together, specifically, the following derivatives. In addition, carbon number attached | subjected to the alkylmethylene group shows carbon number of the alkyl group part of an alkylmethylene group.

In the above formula (1), R ¹ and R ² are both hydrogen atoms, one is a hydrogen atom and the other is a C ₁ -C ₆ alkyl group, both are methylene groups, or both are together Te represents a _C 1 -C ₅ alkyl methylene group. Among these, both are hydrogen atoms, either one is a hydrogen atom and the other is a methyl group, and both are preferably a methylene group.

In the above formula (2), R ³ and R ⁴ are both hydrogen atoms, either one is a hydrogen atom and the other is a C ₁ -C ₆ alkyl group, both together are a methylene group, or both are together Te represents a _C 1 -C ₅ alkyl methylene group. Among these, both are a hydrogen atom, a methyl group, and both are preferably a methylene group.

In the above formulas (1) and (2), when R ¹ and R ² , R ³ and R ⁴ together represent a C ₁ -C ₅ alkylmethylene group, a carbon-carbon double bond in the group The geometric chemistry of (E) or (Z) may be used, or a mixture thereof.

In the above formulas (1) and (2), when R ¹ and R ² , R ³ and R ⁴ are either a hydrogen atom and the other is a C ₁ -C ₆ alkyl group, R ¹ and R The stereochemistry of the carbon atom to be substituted ² is in the foreground according to the steroid stereo notation, and in the case of the above formula (1), the C ₁ -C ₆ alkyl group is on the opposite side of the paper surface. In the case of the above formula (2), the case where the C ₁ -C ₆ alkyl group is in front of the paper surface is the α body, and the one on the opposite side is the β body.

19-nor previtamin D ₃ derivatives of the present invention can be converted to a solvate of their pharmaceutically acceptable as needed. Such solvents include water, methanol, ethanol, 1-propanol, 2-propanol, butanol, t-butanol, acetonitrile, acetone, methyl ethyl ketone, chloroform, ethyl acetate, diethyl ether, Examples thereof include t-butylmethyl ether, benzene, toluene, DMF, DMSO and the like. Particularly preferred are water, methanol, ethanol, 1-propanol, 2-propanol, acetonitrile, acetone, methyl ethyl ketone, and ethyl acetate.

Specific examples of the 19-norprevitamin D ₃ derivative (1) of the present invention include the following.
Compound (1a): 14-epi-1α, 25-dihydroxy-19-norprevitamin D ₃
Compound (1b): 2-methyl-14-epi-1α, 25-dihydroxy-19-norprevitamin D ₃
Compound (1c): 2-methylene-14-epi-1α, 25-dihydroxy-19-norprevitamin D ₃

The 19-norvitamin D ₃ derivative represented by the above formula (1) may be produced by any method, for example, as follows.
When both R ¹ and R ² are hydrogen atoms, they can be produced as shown in Scheme 5. That is, compound (10) (Y. Wu et al., European Journal of Organic Chemistry (Eur. J. Org. Chem.), 3779-3818, 2001) known in the literature is known. Triflate (11) and this and known compound (12a) (LA Sarandes et al., Tetrahedron Letters, 33, 5445-5448, 1992) ), And then the protective group for the hydroxyl group is deprotected to obtain the compound (13a). By performing hydrogen reduction using a Lindlar catalyst, (1) (R ¹ = R ² = hydrogen atom) can be obtained.

In the case where either R ¹ or R ² is a hydrogen atom and the other is a C ₁ -C ₆ alkyl group, both are methylene groups together, or both are C ₁ -C ₅ alkyl methylene groups 6 can be produced. That is, compound (10) (Y. Wu et al., European Journal of Organic Chemistry (Eur. J. Org. Chem.), 3779-3818, 2001) known in the literature is known. After leading to triflate (11) and coupling with (12b), which can be synthesized as shown in Scheme 7, the protecting group of the hydroxyl group is deprotected, and compound (2) (R ¹ and R ² are hydrogen atoms) obtain alkyl methylene group) methylene group or a hydrogen atom and _C 1 -C _5, a. This is subjected to hydrogen reduction under a rhodium catalyst and then hydrogen reduction under a Lindlar catalyst, so that (1) ( ^one of R ¹ and R ² is a hydrogen atom and the other is a C ₁ -C ₆ alkyl group, One of R ¹ and R ² is a hydrogen atom and the other is a C ₁ -C ₅ methylene group, or one of them is a hydrogen atom and the other is a C ₁ -C ₅ alkylmethylene group), and (2) ( One of R ¹ and R ² is a hydrogen atom and the other is a C ₁ -C ₆ alkyl group).

(In Scheme 6 and Scheme 7, R represents a hydrogen atom or a C ₁ -C ₅ alkyl group. In Scheme 7, R ′ represents an aryl group such as a phenyl group, or a lower alkyl group such as a methyl group or an ethyl group. Represents.)

The 19-norprevitamin D ₃ derivative (1) obtained as described above can be converted into a pharmaceutically acceptable solvate as described above, if necessary.
Such a solvate can be obtained by recrystallizing the free compound (1) from the solvent or a mixed solvent containing the solvent.

The osteoporosis therapeutic agent containing the 19-norprevitamin D ₃ derivative of the present invention or a pharmaceutically acceptable solvate thereof as an active ingredient contains carriers, excipients, and other additives that are usually used for formulation. Prepared. The carrier or excipient for the preparation may be either solid or liquid, such as lactose, magnesium stearate, starch, talc, gelatin, agar, pectin, gum arabic, olive oil, sesame oil, cocoa butter, ethylene glycol, etc. The usual thing is mentioned. Administration may be in any form of oral administration such as tablets, pills, capsules, granules, powders, liquids, or parenteral administration such as injections such as intravenous injection and intramuscular injection, suppositories, and transdermal. Good.

The therapeutically effective amount of the active ingredient of the present invention varies depending on the administration route, patient age, sex, and degree of disease, but is usually about 0.001 to 10000 μg / day, and the number of administration is usually 1 to 3 times / day to 1 to 3 times / week, and it is preferable to prepare the preparation so as to satisfy such conditions.
The disease therapeutic agent of the present invention can be used in combination with existing drugs.

EXAMPLES Hereinafter, although an Example demonstrates this invention further in detail, this invention is not limited by this.

[Example 1]
Production of 14-epi-1α, 25-dihydroxy-19-norprevitamin D ₃ (compound (1a))

(1) Diisopropylamine (80 μL, 0.57 mmol) was dissolved in THF (2 mL) and cooled to 0 ° C. n-BuLi (206 μL, 2.77 M hexane solution, 0.57 mmol) was added, and the mixture was stirred at 0 ° C. for 40 minutes. Thereafter, the mixture was cooled to −78 ° C., and a method known from literature (Y. Wu et al., European Journal of Organic Chemistry (Eur. J. Org. Chem.), Pages 3779-3818, 2001) (10) (150 mg, 0.38 mmol) in THF (1 mL) was added and the temperature was raised slowly. After 80 minutes, when it reached room temperature, it was cooled again to −78 ° C., PhN (Tf) ₂ (272 mg, 0.76 mmol) dissolved in THF (2 mL) was added, the temperature was slowly raised, and the mixture was stirred at room temperature overnight. . After 14 hours, the mixture was cooled to 0 ° C., saturated aqueous sodium hydrogen carbonate solution was added, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography (hexane: ethyl acetate = 100: 1-50: 1) to give (11) (122 mg, 61%) as a colorless oil.
¹ H NMR (400 MHz, CDCl ₃ ) δ 5.71 (dt, J = 1.5, 4.2 Hz, 1H), 2.26 (m, 1H), 2.15-2.18 (m, 2H), 1.85-1.99 (m, 2H), 1.28-1.62 (m, 11H), 1.21 (m, 1H), 1.19 (s, 3H), 1.19 (s, 3H), 0.98 (s, 3H), 0.94 (t, J = 8.1 Hz, 9H), 0.93 (s, 3H), 0.56 (q, J = 8.1 Hz, 6H);
¹³ C NMR (100 MHz, CDCl ₃ ) δ 152.5, 120.0, 116.7, 73.4, 51.3, 51.0, 45.5, 44.4, 35.9, 33.8, 33.2, 30.1, 29.9, 28.8, 26.7, 21.6, 21.6, 21.2, 19.5, 7.2 , 6.9;
LRMS (ESI +) m / z 549 (M + Na) ⁺ ;
HRMS (ESI +) calcd for C ₂₅ H ₄₅ O ₄ F ₃ SiNaS, 549.2652, found: 549.2653.

(2) (11) (22 mg, 0.05 mmol) obtained above, the method described in the literature (Salandesses et al., Tetrahedron Letters, 33, 5445-5448) 1992), Compound (12a) (26 mg, 0.06 mmol) dissolved in DMF (0.5 mL), diethylamine (0.5 mL), CuI (4.8 mg, 0.025 mmol), Pd (PPh ₃ ) ₂ (OAc) ₂ (5.6 mg, 0.0075 mmol) was sequentially added, and the mixture was stirred at room temperature for 2 hours. Thereafter, the mixture was ice-cooled, diethyl ether was added, and the mixture was washed with saturated brine. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography (hexane: diethyl ether = 200: 1-100: 1) to give a colorless oil. This was immediately dissolved in THF (2 mL), tetrabutylammonium fluoride (500 μL, 1M THF solution, 0.5 mmol) was added, and the mixture was stirred at room temperature for 14 hours. After cooling to 0 ° C., saturated brine was added, and the mixture was extracted with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography (hexane: ethyl acetate = 1: 1-0: 1) to give (13a) (16 mg, 80%) as a colorless oil.
¹ H NMR (400 MHz, CDCl ₃ ) δ 6.04-6.09 (m, 2H), 4.43-4.48 (m, 1H), 4.12-4.20 (m, 1H), 2.52 (dd, J = 4.2, 17.3 Hz, 1H ), 1.56-2.15 (m, 7H), 1.30-1.60 (m, 10H), 1.20-1.30 (m, 2H), 1.21 (s, 3H), 1.21 (s, 3H), 0.97-1.07 (m, 2H ), 0.93 (d, J = 6.3 Hz, 3H), 0.92 (s, 3H);
¹³ C NMR (100 MHz, CDCl ₃ ) δ 134.0, 133.0, 124.6, 121.7, 90.9, 87.0, 71.1, 65.1, 63.9, 51.8, 51.3, 44.4, 41.2, 39.1, 38.6, 36.0, 34.1, 33.4, 29.4, 29.3 , 28.9, 28.6, 23.1, 21.9, 21.4, 19.4;
[α] _D ²⁵ = + 18.2 (c 0.54, CHCl ₃ );
LRMS (ESI +) m / z 423 (M + Na) ⁺ ;
HRMS (ESI +) calcd for C ₂₆ H ₄₀ O ₃ Na, 423.2870, found: 423.2851.

(3) (13a) (14 mg, 0.035 mmol) obtained above was dissolved in a mixed solvent of methanol (2 mL) and CH ₂ Cl ₂ (1 mL), Quinoline (0.05 mL), Lindlar catalyst (11 mg) Were sequentially added and stirred vigorously under a hydrogen atmosphere for 4.5 hours. Then, it filtered using celite and concentrated under reduced pressure. The crude product obtained by subjecting the obtained crude product to silica gel column chromatography (ethyl acetate) was further separated by preparative chromatography (hexane: ethyl acetate = 1: 4), and (1a) (5 0.5 mg, 39%) as a colorless oil.
¹ H NMR (400 MHz, CDCl ₃ ) δ 5.86 (d, J = 12.5 Hz, 1H), 5.81 (d, J = 12.5 Hz, 1H), 5.55 (m, 1H), 4.56 (m, 1H), 4.07 -4.15 (m, 1H), 2.60 (dd, J = 4.5, 17.1 Hz, 1H), 1.96-2.07 (m, 4H), 1.77-1.94 (m, 4H), 1.26-1.58 (m, 12H), 1.21 (s, 3H), 1.21 (s, 3H), 1.12 (m, 1H), 0.95 (d, J = 6.4 Hz, 3H), 0.93 (s, 3H);
¹³ C NMR (100 MHz, CDCl ₃ ) δ 138.5, 136.6, 133.2, 129.5, 128.1, 125.5, 71.1, 65.7, 64.6, 52.0, 51.5, 44.5, 41.3, 39.8, 37.5, 36.0, 34.3, 33.9, 29.7, 29.2 , 29.1, 28.9, 22.8, 21.9, 20.9, 19.7;
[α] _D ²⁴ = -74.0 (c 0.23, CHCl ₃ );
LRMS (ESI +) m / z 425 (M + Na) ⁺ ;
HRMS (ESI +) calcd for C ₂₆ H ₄₂ O ₃ Na, 425.3026, found: 425.3024.

[Example 2]
2-methyl-14-epi-1α, 25-dihydroxy-19-norprevitamin D ₃ (compound (1b)) and 2-methylene-14-epi-1α, 25-dihydroxy-19-norprevitamin D ₃ ( Production of compound (1c))

(1) Obtained by the method described in the literature (Sicinski et al., Journal of Medicinal Chemistry, Vol. 41, 4462-4694, 1998) (15) (R = hydrogen atom) (600 mg, 1.39 mmol) was dissolved in pyridine (3 mL), POCl ₃ (195 μL, 2.09 mmol) was added, and the mixture was stirred for 48 hours. Thereafter, the mixture was cooled to 0 ° C., water was added, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography (hexane: ethyl acetate = 100: 1-50: 1) to give (16) (R = hydrogen atom) (488 mg, 85%) as a colorless oil. Obtained.
¹ H NMR (400 MHz, CDCl ₃ ) δ 6.75 (m, 1H), 5.08 (s, 1H), 5.02 (s, 1H), 4.91 (m, 1H), 4.56 (dd, J = 5.1, 5.6 Hz, 1H), 3.73 (s, 3H), 2.67 (ddd, J = 1.9, 5.1, 17.6 Hz, 1H), 2.31 (ddd, J = 1.7, 5.6, 17.6 Hz, 1H), 0.91 (s, 9H), 0.89 (s, 9H), 0.12 (s, 3H), 0.09 (s, 3H), 0.07 (s, 3H), 0.06 (s, 3H);
¹³ C NMR (100 MHz, CDCl ₃ ) δ 167.1, 148.8, 139.5, 129.2, 108.1, 69.5, 69.0, 51.8, 36.8, 25.9, 25.8, 18.3, 18.2, -4.7, -4.7, -4.8, -4.9;
LRMS (ESI +) m / z 435 (M + Na) ⁺ ;
HRMS (ESI +) calcd for C ₂₁ H ₄₀ O ₄ Si ₂ Na, 435.2363, found: 435.2363.

(2) (16) (R = hydrogen atom) (330 mg, 0.80 mmol) obtained above was dissolved in toluene (3 mL) and cooled to 0 ° C. DIBAL-H (2.42 mL, 0.99 M toluene solution, 2.40 mmol) was added, and the mixture was stirred at room temperature for 42 hours. Thereafter, the mixture was cooled to 0 ° C., water (0.1 mL), 15% aqueous sodium hydroxide solution (0.1 mL) and water (0.3 mL) were sequentially added, and the mixture was stirred at room temperature overnight. Magnesium sulfate was added to the reaction mixture, and the mixture was stirred, filtered through celite, and concentrated under reduced pressure. The resulting crude product was separated by silica gel column chromatography (hexane: ethyl acetate = 8: 1-6: 1) to give (17) (R = hydrogen atom) (267 mg, 87%) as a colorless oil. It was.
¹ H NMR (400 MHz, CDCl ₃ ) δ 5.64 (m, 1H), 5.08 (s, 1H), 4.97 (s, 1H), 4.77 (m, 1H), 4.60 (dd, J = 5.3, 6.7 Hz, 1H), 3.99 (s, 1H), 2.40 (dd, J = 5.3, 16.7 Hz, 1H), 2.06 (dd, J = 6.7, 16.7 Hz, 1H), 0.90 (s, 9H), 0.90 (s, 9H ), 0.10 (s, 3H), 0.08 (s, 3H), 0.06 (s, 3H), 0.05 (s, 3H);
¹³ C NMR (100 MHz, CDCl ₃ ) δ 150.4, 138.2, 124.1, 107.4, 69.9, 69.1, 66.0, 38.3, 25.9, 25.9, 25.8, 25.7, 18.4, 18.3, -4.5, -4.6, -4.8, -4.9 ;
LRMS (ESI +) m / z 407 (M + Na) ⁺ ;
HRMS (ESI +) calcd for C ₂₀ H ₄₀ O ₃ Si ₂ Na, 407.2414, found: 407.2418.

(3) (17) (R = hydrogen atom) (470 mg, 1.22 mmol) obtained above was dissolved in methylene chloride (6 mL), and MS4A (610 mg), NMO (215 mg, 1.83 mmol), TPAP ( 21 mg, 0.061 mmol) was sequentially added, and the mixture was stirred at room temperature for 2 hours. Thereafter, the mixture is filtered using Celite and concentrated under reduced pressure. The resulting crude product is subjected to neutral silica gel column chromatography (hexane: ethyl acetate = 100: 1-50: 1) to give an aldehyde as a colorless oil. Obtained. This was immediately used for the following reaction. TMSCHN ₂ (686 μL, 2.0 M ether solution, 1.37 mmol) was dissolved in THF (4 mL) and cooled to −78 ° C. n-BuLi (479 μL, 2.77 M hexane solution, 1.33 mmol) was added and stirred for 30 minutes. An aldehyde dissolved in THF (1 mL) was added, and the temperature was slowly raised. After 2 hours, when the temperature reached 0 ° C., the mixture was further stirred at that temperature for 15 minutes. Thereafter, a saturated aqueous ammonium chloride solution was added, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography (hexane: ethyl acetate = 100: 1-50: 1) to give (12b) (R = hydrogen atom) (229 mg, 50% (2 steps)) colorless. Obtained as an oil.
¹ H NMR (400 MHz, CDCl ₃ ) δ 6.05 (m, 1H), 5.09 (s, 1H), 5.01 (s, 1H), 4.80 (m, 1H), 4.56 (dd, J = 5.4, 6.6 Hz, 1H), 2.85 (s, 1H), 2.52 (ddd, J = 1.5, 5.4, 17.1 Hz, 1H), 2.22 (ddd, J = 1.6, 6.6, 17.1 Hz, 1H), 0.90 (s, 9H), 0.90 (s, 9H), 0.10 (s, 3H), 0.08 (s, 3H), 0.08 (s, 3H), 0.06 (s, 3H);
¹³ C NMR (100 MHz, CDCl ₃ ) δ 148.9, 137.0, 119.8, 108.0, 83.9, 76.2, 69.5, 68.8, 41.3, 25.9, 25.8, 18.3, 18.3, -4.6, -4.7, -4.8, -4.9;
LRMS (ESI +) m / z 401 (M + Na) ⁺ ;
HRMS (ESI +) calcd for C ₂₁ H ₃₈ O ₂ Si ₂ Na, 401.2303, found: 401.2304.

(4) (12b) (R = hydrogen atom) (162 mg, 0.428 mmol), (11) (150 mg, 0.285 mmol) obtained in Example 1 (1) was dissolved in DMF (1.5 mL). , Diethylamine (1.5 mL), CuI (27 mg, 0.143 mmol), Pd (PPh ₃ ) ₂ (OAc) ₂ (32 mg, 0.0428 mmol) were sequentially added, and the mixture was stirred at room temperature for 90 minutes. Thereafter, the mixture was ice-cooled, diethyl ether was added, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography (hexane: diethyl ether = 200: 1-100: 1) to give a colorless oil. This was immediately dissolved in THF (4 mL), tetrabutylammonium fluoride (2.85 mL, 1M THF solution, 2.85 mmol) was added, and the mixture was stirred at room temperature for 19 hours. After cooling to 0 ° C., saturated brine was added, and the mixture was extracted with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography (hexane: ethyl acetate = 1: 1-1: 3) to give (2) (R ¹ / R ² = methylene group) (100 mg, 85%). Obtained as a colorless oil.
¹ H NMR (400 MHz, CDCl ₃ ) δ 6.07-6.09 (m, 1H), 6.03-6.05 (m, 1H), 5.21 (d, J = 1.0 Hz, 1H), 5.19 (d, J = 1.0 Hz, 1H), 4.83 (m, 1H), 4.64 (dd, J = 5.1, 7.1 Hz, 1H), 2.66 (ddd, J = 1.5, 5.1, 17.1 Hz, 1H), 2.28 (ddd, J = 1.5, 7.1, 17.1 Hz, 1H), 1.93-2.09 (m, 4H), 1.78-1.82 (m, 1H), 1.29-1.57 (m, 10H), 1.25 (m, 1H), 1.21 (s, 3H), 1.21 (s , 3H), 1.06 (m, 1H), 0.95 (d, J = 6.5 Hz, 3H), 0.91 (s, 3H);
¹³ C NMR (100 MHz, CDCl ₃ ) δ 148.8, 134.1, 132.4, 124.5, 122.2, 109.2, 91.3, 86.6, 71.1, 69.1, 68.2, 51.8, 51.3, 44.5, 41.3, 40.4, 36.1, 34.2, 33.4, 29.6 , 29.5, 27.3, 27.3, 23.2, 22.0, 21.4, 19.5 .;
[α] _D ²⁵ = + 19.2 (c 0.85, CHCl ₃ );
LRMS (ESI +) m / z 435 (M + Na) ⁺ ;
HRMS (ESI +) calcd for C ₂₇ H ₄₀ O ₃ Na, 435.2870, found: 435.2859.

(5) (PPh ₃ ) ₃ RhCl (105 mg, 0.114 mmol) was dissolved in benzene (2 mL) and stirred under a hydrogen stream for 40 minutes. A solution obtained by dissolving (2) (R ¹ / R ² = methylene group) (47 mg, 0.114 mmol) obtained in the above in a mixed solvent of benzene (2 mL) and methylene chloride (1 mL) was added to the reaction solution, and hydrogen was added. The mixture was vigorously stirred for 2 hours under an air stream. Thereafter, the reaction mixture was concentrated under reduced pressure, and the crude product obtained was subjected to silica gel column chromatography (dichloromethane: methanol = 20: 1-15: 1) to immediately obtain methanol (2 mL) and methylene chloride (1 mL). It melt | dissolved in the mixed solvent, Quinoline (0.05 mL) and Lindlar catalyst (18 mg) were added one by one to this, and it stirred vigorously under hydrogen atmosphere for 15 hours. Then, it filtered using celite and concentrated under reduced pressure. The crude product obtained by subjecting this to silica gel column chromatography (hexane: ethyl acetate = 1: 1-1: 3) was further separated by preparative chromatography (hexane: ethyl acetate = 1: 1), The isomer mixture (1b) was obtained as a colorless oil. This isomer mixture was separated by HPLC (CH ₃ CN: H ₂ O = 9: 1) to obtain (1b) -1 (12 mg, 25%), (1b) -2 (15 mg, 32%). In addition, a small amount of two isomers (2) -1 and (2) -2 of (1c) and (2) (R ³ / R ⁴ = hydrogen atom / methyl group) were obtained. In addition, (1b) -1 and (1b) -2 are based on the asymmetry of the carbon atom to which the methyl group corresponding to R ¹ / R ² of the general formula (1) is bonded, (2) -1 and (2 ) -2 is based on the asymmetry of the carbon atom to which the methyl group corresponding to R ³ / R ⁴ in the general formula (2) is bonded.

(1b) -1:
¹ H NMR (400 MHz, CDCl ₃ ) δ 5.88 (d, J = 12.2 Hz, 1H), 5.76 (d, J = 12.2 Hz, 1H), 5.72 (m, 1H), 5.53 (m, 1H), 4.08 (m, 1H), 4.04 (m, 1H), 2.25-2.57 (m, 1H), 2.15 (dd, J = 4.9, 17.8 Hz, 1H), 1.97-2.05 (m, 3H), 1.65-1.88 (m , 3H), 1.32-1.55 (m, 13H), 1.23 (s, 3H), 1.23 (s, 3H), 1.07-1.13 (m, 1H), 1.09 (d, J = 6.8 Hz, 3H), 0.95 ( d, J = 6.4 Hz, 3H), 0.92 (s, 3H);
¹³ C NMR (100 MHz, CDCl ₃ ) δ 138.5, 135.4, 132.9, 129.8, 128.7, 125.3, 77.2, 71.2, 69.3, 51.9, 51.6, 44.5, 41.2, 41.1, 36.1, 35.6, 34.3, 33.9, 29.8, 29.1 , 29.0, 22.8, 21.9, 20.8, 19.7, 13.1;
[α] _D ²⁴ = -120.9 (c 0.23, CHCl ₃ );
LRMS (ESI +) m / z 439 (M + Na) ⁺ ;
HRMS (ESI +) calcd for C ₂₇ H ₄₄ O ₃ Na, 439.3183, found: 439.3193.
(1b) -2:
¹ H NMR (400 MHz, CDCl ₃ ) δ 5.83 (s, 1H), 5.83 (s, 1H), 5.74 (m, 1H), 5.56 (m, 1H), 4.26 (m, 1H), 3.77 (m, 1H), 2.64 (dd, J = 4.9, 17.6 Hz, 1H), 1.95-2.08 (m, 4H), 1.74-1.87 (m, 3H), 1.28-1.59 (m, 14H), 1.22 (s, 3H) , 1.22 (s, 3H), 1.11 (d, J = 6.8 Hz, 3H), 0.95 (d, J = 6.4 Hz, 3H), 0.93 (s, 3H);
¹³ C NMR (100 MHz, CDCl ₃ ) δ 138.6, 136.9, 133.3, 129.3, 128.0, 125.4, 77.2, 71.1, 69.2, 68.8, 52.0, 51.4, 44.4, 41.3, 40.7, 37.1, 35.9, 34.2, 33.9, 29.7 , 29.1, 29.1, 28.8, 22.8, 21.8, 20.8, 19.6, 12.3;
[α] _D ²⁴ = -39.5 (c 0.62, CHCl ₃ );
LRMS (ESI +) m / z 439 (M + Na) ⁺ ;
HRMS (ESI +) calcd for C ₂₇ H ₄₄ O ₃ Na, 439.3183, found: 439.3176.
(1c):
¹ H NMR (400 MHz, CDCl ₃ ) δ 5.88 (d, J = 12.2 Hz, 1H), 5.81 (d, J = 12.2 Hz, 1H), 5.75 (m, 1H), 5.54 (m, 1H), 5.19 (s, 1H), 5.16 (s, 1H), 4.82 (m, 1H), 4.58 (m, 1H), 2.73 (dd, J = 5.2, 16.7 Hz, 1H), 2.18 (dd, J = 7.3, 16.7 Hz, 1H), 1.96-2.06 (m, 4H), 1.76-1.87 (m, 2H), 1.27-1.56 (m, 9H), 1.22 (s, 3H), 1.22 (s, 3H), 1.07-1.14 ( m, 1H), 0.95 (d, J = 7.4 Hz, 3H), 0.88 (s, 3H);
¹³ C NMR (100 MHz, CDCl ₃ ) δ 138.5, 137.2, 133.5, 129.2, 128.1, 125.7, 108.5, 77.2, 71.2, 69.5, 58.7, 52.0, 51.5, 44.4, 41.2, 39.1, 35.9, 34.2, 33.9, 29.7 , 29.7, 29.1, 28.8, 22.8, 21.8, 20.8, 19.6;
[α] _D ²⁴ = -209.0 (c 0.27, CHCl ₃ );
LRMS (ESI +) m / z 437 (M + Na) ⁺ ;
HRMS (ESI +) calcd for C ₂₇ H ₄₂ O ₃ Na, 437.3026, found: 437.3010.
(2) -1:
¹ H NMR (400 MHz, CDCl ₃ ) δ 6.07-6.09 (m, 1H), 6.03-6.05 (m, 1H), 4.05-4.08 (m, 2H), 2.50 (m, 1H), 2.24 (m, 1H ), 1.93-2.09 (m, 4H), 1.95-2.10 (m, 4H), 1.71-1.84 (m, 2H), 1.28-1.57 (m, 13H), 1.22 (s, 3H), 1.22 (s, 3H ), 1.09 (d, J = 7.1 Hz, 3H), 1.06 (m, 1H), 0.93 (d, J = 6.4 Hz, 3H), 0.92 (s, 3H);
¹³ C NMR (100 MHz, CDCl ₃ ) δ 133.8, 133.4, 124.6, 120.5, 90.7, 87.0, 77.2, 70.6, 68.7, 51.8, 51.3, 44.5, 41.3, 40.8, 37.1, 36.1, 34.2, 33.5, 29.8, 29.5 , 29.3, 29.0, 28.6, 23.1, 22.0, 21.4, 19.5, 13.6; [α] _D ²⁵ = + 28.5 (c 0.15, CHCl ₃ );
LRMS (ESI +) m / z 437 (M + Na) ⁺ ;
HRMS (ESI +) calcd for C ₂₇ H ₄₂ O ₃ Na, 437.3026, found: 437.3017.
(2) -2:
¹ H NMR (400 MHz, CDCl ₃ ) δ 6.06-6.10 (m, 2H), 4.28 (m, 1H), 4.28 (m, 1H), 3.85 (m, 1H), 2.58 (dd, J = 5.4, 17.6 Hz, 1H), 2.11-2.17 (m, 1H), 1.95-2.09 (m, 4H), 1.75-1.85 (m, 2H), 1.20-1.60 (m, 13H), 1.22 (s, 1H), 1.22 ( s, 3H), 1.10 (d, J = 6.8 Hz, 3H), 1.04-1.09 (m, 1H), 0.94 (d, J = 6.3 Hz, 3H), 0.92 (s, 3H);
¹³ C NMR (100 MHz, CDCl ₃ ) δ 133.9, 132.8, 124.5, 121.9, 90.9, 86.9, 77.2, 71.1, 68.7, 68.2, 51.8, 51.3, 44.6, 41.3, 40.2, 38.3, 36.1, 34.2, 33.5, 29.8 , 29.3, 29.0, 28.6, 23.1, 22.0, 21.4, 19.5, 12.2;
[α] _D ²⁵ = -2.2 (c 0.69, CHCl ₃ );
LRMS (ESI +) m / z 437 (M + Na) ⁺ ;
HRMS (ESI +) calcd for C ₂₇ H ₄₂ O ₃ Na, 437.3026, found: 437.3033.

[Example 3]
VDR transcriptional activity in human osteoblasts (HOS cells) (1) Using a pGL3 vector (promega) as a reporter vector, upstream of the luciferase gene, a method known in the literature (Ozono et al., The Journal of Biological) The sequence of the human osteocalcin gene promoter portion obtained in Chemistry (The Journal of Biological Chemistry), 265, 21881-21888, 1990) was cloned from cDNA obtained from HOS cells (obtained from ATCC) and incorporated. The expression vector was constructed by inserting DNA sequences encoding human VDR and human RXR into a pCDNA3 vector (Invitrogen). HOS cells are 10% F
The cells were cultured in a DMEM medium containing BS at 37 ° C. and 5% CO ₂ and subcultured every 2 or 3 days.

(2) The subcultured cells were collected by centrifugation, dispersed in serum-free, phenol red-free DMEM medium at a density of 4 × 10 ⁵ cells / ml, and seeded in a 96-well plate at 0.1 mL / well. did. To this system, 0.05 mL of each vector described in (1) was added per well using Lipofectamine 2000 (Invitrogen) reagent. After incubating at 37 ° C. for 3 hours, 2 μL of 1 × 10 ⁻¹³ to 1 × 10 ⁻⁷ M test compound ethanol solution or ethanol as a control was added to each well. After incubation at 37 ° C. for 24 hours, the medium was removed, washed once with PBS (−), and then luciferase activity was measured with a luminometer (Berthold) using DualGlo-Luciferase Assay kit (Promega). When the activity of the test compound is plotted against its treatment concentration, the activity of 1 × 10 ⁻⁷ M 1α, 25-dihydroxyvitamin D ₃ treatment is 100%, and the control ethanol treatment activity is 0%. , The treatment concentration of the test compound to be 50% was calculated as an EC ₅₀ value (nM). The results are shown in Table 1. Moreover, the activity of 19th-position natural type compound (A), (B), (C) corresponding as a comparative example was measured. The results are shown in Table 1.

(A) 14-epi-1α, 25-dihydroxy previtamin D ₃
(B) 2β-methyl-14-epi-1α, 25-dihydroxy previtamin D ₃
(C) 2α-methyl-14-epi-1α, 25-dihydroxyprevitamin D ₃
From this result, the compounds of the present invention, compared to known previtamin D ₃ derivative (8), was found to have a significantly stronger transcriptional activity.

19-nor previtamin D ₃ derivatives of the present invention is used as an active ingredient of osteoporosis therapeutic agents.

Claims (9)

Represented by 19-nor previtamin D ₃ derivative or a pharmaceutically acceptable solvate thereof by the following formula (1).
[Wherein, R ¹ and R ² are both hydrogen atoms, one is a hydrogen atom and the other is a C ₁ -C ₆ alkyl group, both are methylene groups, or both are C ₁ -C represents a ₅ alkyl methylene group. ] ^2. The 19-norprevitamin D ₃ derivative or a pharmaceutically acceptable solvate thereof according to claim 1, wherein R ¹ and R ^{2 in} the formula (1) are both hydrogen atoms. ^2. The 19-norprevitamin D ₃ derivative or a pharmaceutically acceptable solvate thereof according to claim 1, wherein one of R ¹ and R ^{2 in} the formula (1) is a hydrogen atom and the other is a methyl group. . ^2. The 19-norprevitamin D ₃ derivative or a pharmaceutically acceptable solvate thereof according to claim 1, wherein R ¹ and R ^{2 in} the formula (1) are both methylene groups. Osteoporosis agent containing 19-nor previtamin D ₃ derivative or a pharmaceutically acceptable solvate thereof as an active ingredient according to any one of claims 1 to 4. A production intermediate of a 19-norprevitamin D ₃ derivative represented by the following formula (2).
[Wherein, R ³ and R ⁴ are both hydrogen atoms, one is a hydrogen atom and the other is a C ₁ -C ₆ alkyl group, both are methylene groups, or both are C ₁ -C represents a ₅ alkyl methylene group. ] The intermediate for producing a 19-norprevitamin D ₃ derivative according to claim 6, wherein R ³ and R ^{4 in} the formula (2) are both hydrogen atoms. The intermediate for producing a 19-norprevitamin D ₃ derivative according to claim 6, wherein one of R ³ and R ^{4 in} the formula (2) is a hydrogen atom and the other is a methyl group. The intermediate for producing a 19-norprevitamin D ₃ derivative according to claim 6, wherein R ³ and R ^{4 in} the formula (2) are both methylene groups.