JP7572027B2 - Compounds with a secosteroid structure - Google Patents

Description

The present invention relates to a compound having a secosteroid structure, a composition containing the same, and a method for producing the same, and in particular to a novel vitamin D derivative compound.

Vitamin D is known to be involved in intestinal absorption of calcium and phosphorus, bone metabolism, cell differentiation and proliferation, and immunity. It is also known that a deficiency of vitamin D can cause rickets, osteomalacia, or reduced bone mass in pregnant women and fetuses, which are related to bone formation, and can increase the risk of elderly people becoming dependent on care or becoming frail. Vitamin D can be synthesized in the skin when exposed to sunlight, but in high latitude areas, the hours of sunlight are short, so sufficient amounts of vitamin D may not be synthesized. The amount of vitamin D synthesis can also decrease due to lack of sunlight, excessive whitening care, or the use of inhibitors of the cholesterol synthesis pathway. Meanwhile, foods that contain vitamin D are limited to seafood, egg yolks, and mushrooms, and it is difficult to ingest enough of it from a normal diet.

Vitamins D2 to D7 are known as vitamin D, and vitamins D2 to D4 are commercially available, but vitamins D5 to D7 and their precursors are not commercially available. Non-Patent Document 1 describes the simultaneous synthesis of vitamins D2 and D4 to D7 from a commercially available plant sterol mixture, and the synthesis of vitamin D3 from pure cholesterol.

Komba, S., et al., Metabolites, 2019, 9(6), 107, doi: 10.3390/metabo9060107

It is desirable to search for unknown naturally occurring vitamin D and increase the opportunities to consume foods containing vitamin D to alleviate vitamin D deficiency or to utilize it as a functional ingredient, but in order to do so, it is necessary to synthesize and prepare a standard substance. Therefore, the present invention aims to provide a novel compound that has a secosteroid structure, which is a common structure of vitamin D.

（Ｉ）
（式中、
Ｒ¹は、ＯＨ基、Ｏ－Ｃ₂～C₁₈アシル基、又はグリコシド基であり、
Ｒ²は、Ｈ、ＯＨ基、又はＯ－Ｃ₂～C₁₈アシル基であり、
Ｒ³及びＲ⁴は、それぞれ独立して、Ｈ、メチル基、若しくはＯＨ基であるか、又は、Ｒ³及びＲ⁴は、これらが結合している炭素原子と一緒にメチレン基を形成し、
実線及び破線からなる二重線は、単結合又は二重結合を表す）
で表される化合物又はその立体異性体。
〔２〕実線及び破線からなる前記二重線が、単結合を表す、前記〔１〕に記載の化合物又はその立体異性体。
〔３〕Ｒ¹がＯＨ基であり、Ｒ²がＨである、前記〔１〕又は〔２〕に記載の化合物又はその立体異性体。
〔４〕Ｒ¹がＯＨ基であり、Ｒ²がＯＨ基である、前記〔１〕又は〔２〕に記載の化合物又はその立体異性体。
〔５〕Ｒ³がメチル基であり、かつＲ⁴がＨであるか、又は、Ｒ³及びＲ⁴が、これらが結合している炭素原子と一緒にメチレン基を形成する、前記〔１〕～〔４〕のいずれか一項に記載の化合物又はその立体異性体。
〔６〕Ｒ³がメチル基であり、かつＲ⁴がＯＨである、前記〔１〕～〔４〕のいずれか一項に記載の化合物又はその立体異性体。
〔７〕以下の式：

のいずれかで表される化合物又はその立体異性体。
〔８〕前記〔１〕～〔７〕のいずれか一項に記載の化合物又はその立体異性体を含む組成物。
〔９〕一般式Ｉ：

（Ｉ）
（式中、
Ｒ¹は、ＯＨ基、Ｏ－Ｃ₂～C₁₈アシル基、又はグリコシド基であり、
Ｒ²は、Ｈ、ＯＨ基、又はＯ－Ｃ₂～C₁₈アシル基であり、
Ｒ³及びＲ⁴は、それぞれ独立して、Ｈ、メチル基、若しくはＯＨ基であるか、又は、Ｒ³及びＲ⁴は、これらが結合している炭素原子と一緒にメチレン基を形成し、
実線及び破線からなる二重線は、単結合又は二重結合を表す）
で表される化合物又はその立体異性体の製造方法であって、
一般式ＩＩ：

（ＩＩ）
（式中の記号は一般式Ｉのものと同じ）
で表される化合物を、一般式ＩＩＩ：

（式中の記号は一般式Ｉのものと同じ）
で表される化合物に変換する工程と、
一般式ＩＩＩで表される前記化合物を、一般式Ｉで表される前記化合物に変換する工程と、
を含む製造方法。 As a result of intensive research aimed at solving the above problems, the present inventors have found a novel compound having a characteristic secosteroid structure in its terminal structure, and have completed the present invention. That is, the present invention provides the following compound, a composition containing the same, and a method for producing the same.
[1] General formula I:

(I)
(Wherein,
^R1 is an OH group, an O— _C2 - _C18 acyl group, or a glycoside group;
^R2 is H, an OH group, or an O- _C2 - _C18 acyl group;
^R3 and ^R4 are each independently H, a methyl group, or an OH group, or ^R3 and ^R4 together with the carbon atom to which they are attached form a methylene group;
A double line consisting of a solid line and a dashed line represents a single bond or a double bond.
A compound represented by the formula:
[2] The compound or stereoisomer thereof according to [1] above, wherein the double line consisting of a solid line and a dashed line represents a single bond.
[3] The compound or stereoisomer thereof according to [1] or [2] above, wherein R ¹ is an OH group and R ² is H.
[4] The compound or stereoisomer thereof according to [1] or [2] above, wherein R ¹ is an OH group and R ² is an OH group.
[5] The compound or stereoisomer thereof according to any one of [1] to [4] above, wherein R ³ is a methyl group and R ⁴ is H, or R ³ and R ⁴ together with the carbon atom to which they are bonded form a methylene group.
[6] The compound or stereoisomer thereof according to any one of the above [1] to [4], wherein R ³ is a methyl group and R ⁴ is OH.
[7] A compound represented by the following formula:

or a stereoisomer thereof.
[8] A composition comprising the compound according to any one of [1] to [7] above or a stereoisomer thereof.
[9] General formula I:

(I)
(Wherein,
^R1 is an OH group, an O— _C2 - _C18 acyl group, or a glycoside group;
^R2 is H, an OH group, or an O- _C2 - _C18 acyl group;
^R3 and ^R4 are each independently H, a methyl group, or an OH group, or ^R3 and ^R4 together with the carbon atom to which they are attached form a methylene group;
A double line consisting of a solid line and a dashed line represents a single bond or a double bond.
A method for producing a compound represented by the formula:
General Formula II:

(II)
(The symbols in the formula are the same as those in general formula I.)
The compound represented by the general formula III:

(The symbols in the formula are the same as those in general formula I.)
and converting the compound represented by the formula:
converting said compound of general formula III to said compound of general formula I;
A manufacturing method comprising:

According to the present invention, a novel vitamin D derivative compound is provided. This expands the variety of available vitamin D derivative compounds.

The thin-layer chromatograph of the fractions obtained by silica column chromatography is shown in Fig. 1. The area enclosed in a square was collected as fraction a. The thin-layer chromatograph of the fractions obtained by silica column chromatography is shown in Fig. 1. The area enclosed in a square was collected as fraction b. The thin-layer chromatograph of the fractions obtained by silica column chromatography is shown in Fig. 1. The area enclosed in a box was collected as fraction c. The high performance liquid chromatograph of fraction c is shown.

（Ｉ）
で表される。ここで、実線及び破線からなる二重線は、単結合又は二重結合、好ましくは単結合を表す。本発明の化合物又はその立体異性体は、ビタミンＤと共通する構造を基本骨格として有しているため、生体内ではビタミンＤと同様の代謝を受け、同様の作用を奏すると期待される。なお、ビタミンＤは、肝臓においてセコステロイド構造の２５位の水酸化を受け、その後腎臓において当該構造の１α位の水酸化を受けて、活性型ビタミンＤに変換されることが知られている。 The present invention will now be described in further detail.
The present invention relates to compounds having a secosteroid structure or a stereoisomer thereof, the chemical structure of which is represented by the general formula I:

(I)
Here, the double line consisting of a solid line and a broken line represents a single bond or a double bond, preferably a single bond. The compound of the present invention or a stereoisomer thereof has a structure common to vitamin D as a basic skeleton, and is therefore expected to be metabolized in the same manner as vitamin D in vivo and to exert the same effect. It is known that vitamin D is hydroxylated at the 25-position of the secosteroid structure in the liver, and then hydroxylated at the 1α-position of the structure in the kidney, to be converted into active vitamin D.

R ¹ is an OH group, an O-C ₂ -C ₁₈ acyl group, or a glycoside group. The acyl group portion may be linear or branched. The acyl group portion may be, for example, -C(=O)-Q ¹ , where Q ¹ is a saturated hydrocarbon chain of C ₁ -C ₁₇ , C ₁ -C ₁₁ , or C ₁ -C ₅ , or an unsaturated hydrocarbon chain of C ₂ -C ₁₇ , C ₂ -C ₁₁ , or C ₂ -C _5. The ester bond and glycoside bond formed when ^{R 1} is an O-C ₂ -C ₁₈ acyl group or a glycoside group may be hydrolyzed in vivo, and as a result of the hydrolysis, a compound having a hydroxyl group, i.e., a compound in which R ¹ is an OH group, may be generated.

R ² is H, an OH group, or an O-C ₂ -C ₁₈ acyl group. The acyl group portion may be linear or branched. The acyl group portion may be, for example, -C(=O)-Q ² , where Q ² is a saturated hydrocarbon chain of C ₁ -C ₁₇ , C ₁ -C ₁₁ , or C ₁ -C ₅ , or an unsaturated hydrocarbon chain of C ₂ -C ₁₇ , C ₂ -C ₁₁ , or C ₂ -C _5. The ester bond formed when ^{R 2} is an O-C ₂ -C ₁₈ acyl group may be hydrolyzed in vivo, and as a result of the hydrolysis, a compound having a hydroxyl group, i.e., a compound in which R ² is an OH group, may be produced. In addition, a compound in which R ² is H may be hydroxylated in the kidney to produce a compound in which R ² is an OH group. In some embodiments, R ¹ and R ² are H, or R ¹ is H and R ² is OH.

R ³ and R ⁴ are each independently H, a methyl group, or an OH group, or R ³ and R ⁴ form a methylene group together with the carbon atom to which they are attached. In some embodiments, R ³ and R ⁴ are not simultaneously methyl groups, for example, a compound in which R ³ or R ⁴ is H or a compound in which R ³ and R ⁴ form a methylene group together with the carbon atom to which they are attached can be hydroxylated in the liver to produce a compound in which R ³ or R ⁴ is an OH group. In some embodiments, R ³ is a methyl group and R ⁴ is H, or R ³ and R ⁴ form a methylene group together with the carbon atom to which they are attached. In some embodiments, R ³ is a methyl group and R ⁴ is OH.

In certain embodiments, the compound of the present invention is represented by a formula selected from the following group, or is a stereoisomer thereof:

The compounds of the present invention can be synthesized from the corresponding precursors by a conventional method. For example, the compounds of the present invention (compounds 3 and 4) may be synthesized from compound 1 having a steroid skeleton as a starting material according to the following scheme. Compounds 3 and 4 may be produced as a mixture and then each may be isolated.
_Ac2O = acetic anhydride
Pyr = pyridine
NBS = N-bromosuccinimide
CYH = cyclohexane
BHA = 3-tert-butyl-4-hydroxyanisole
_Bu4NF = Tetrabutylammonium fluoride
NaOMe = sodium methylate
THF = Tetrahydrofuran
MeOH = Methanol
_CH2Cl2 = _{dichloromethane}
rt = room temperature

The compounds of the present invention may be useful as standard substances for use in the search for vitamin D derivative compounds. Based on their activity as vitamin D, the compounds of the present invention may also be useful as active ingredients for improving or preventing vitamin D deficiency, diseases or symptoms caused by vitamin D deficiency, or diseases treated with vitamin D. The diseases or symptoms caused by vitamin D deficiency or diseases treated with vitamin D may be, but are not limited to, hypocalcemia, hypophosphatemia, metabolic bone diseases in which it is desired to maintain or increase bone mass (such as rickets, osteomalacia, and osteoporosis), and cancer. The compounds of the present invention may exhibit a different activity spectrum from known compounds with respect to the broad physiological effects that vitamin D can exert, thereby expanding the variety of available vitamin D derivative compounds.

（Ｉ）
（式中、
Ｒ¹は、ＯＨ基、Ｏ－Ｃ₂～C₁₈アシル基、又はグリコシド基であり、
Ｒ²は、Ｈ、ＯＨ基、又はＯ－Ｃ₂～C₁₈アシル基であり、
Ｒ³及びＲ⁴は、それぞれ独立して、Ｈ、メチル基、若しくはＯＨ基であるか、又は、Ｒ³及びＲ⁴は、これらが結合している炭素原子と一緒にメチレン基を形成し、
実線及び破線からなる二重線は、単結合又は二重結合を表す）
で表される化合物又はその立体異性体を含む組成物に関している。本発明の組成物は、ビタミンＤ誘導体化合物の探索に用いるための試薬として有用であり得る。また、本発明の組成物は、医薬、食品、又は飼料としても有用であり得る。 In another aspect, the present invention provides compounds of general formula I:

(I)
(Wherein,
^R1 is an OH group, an O— _C2 - _C18 acyl group, or a glycoside group;
^R2 is H, an OH group, or an O- _C2 - _C18 acyl group;
^R3 and ^R4 are each independently H, a methyl group, or an OH group, or ^R3 and ^R4 together with the carbon atom to which they are attached form a methylene group;
A double line consisting of a solid line and a dashed line represents a single bond or a double bond.
The present invention relates to a composition comprising a compound represented by the formula (I) or a stereoisomer thereof. The composition of the present invention may be useful as a reagent for use in searching for vitamin D derivative compounds. The composition of the present invention may also be useful as a medicine, food, or feed.

The composition of the present invention may further contain any buffer or additive commonly used in the art. When the composition of the present invention is in the form of a medicine, food, or feed, it may further contain other active ingredients and/or inactive ingredients as long as they do not interfere with the action of the compound contained therein. Examples of the inactive ingredients that may be used include excipients, disintegrants, binders, lubricants, dispersants, buffers, solvents, bulking agents, thickeners, flavorings, preservatives, and stabilizers.

（Ｉ）
（式中、
Ｒ¹は、ＯＨ基、Ｏ－Ｃ₂～C₁₈アシル基、又はグリコシド基であり、
Ｒ²は、Ｈ、ＯＨ基、又はＯ－Ｃ₂～C₁₈アシル基であり、
Ｒ³及びＲ⁴は、それぞれ独立して、Ｈ、メチル基、若しくはＯＨ基であるか、又は、Ｒ³及びＲ⁴は、これらが結合している炭素原子と一緒にメチレン基を形成し、
実線及び破線からなる二重線は、単結合又は二重結合を表す）
で表される化合物又はその立体異性体の製造方法に関している。本発明の製造方法は、
一般式ＩＩ：

（ＩＩ）
（式中の記号は一般式Ｉのものと同じ）
で表される化合物を、一般式ＩＩＩ：

（式中の記号は一般式Ｉのものと同じ）
で表される化合物に変換する工程と、
一般式ＩＩＩで表される前記化合物を、一般式Ｉで表される前記化合物に変換する工程と、を含んでいる。場合によっては、一般式ＩＩで表される前記化合物及び一般式ＩＩＩで表される前記化合物の置換基は、任意の保護基によって保護されていてもよい。 In another aspect, the present invention provides compounds of general formula I:

(I)
(Wherein,
^R1 is an OH group, an O— _C2 - _C18 acyl group, or a glycoside group;
^R2 is H, an OH group, or an O- _C2 - _C18 acyl group;
^R3 and ^R4 are each independently H, a methyl group, or an OH group, or ^R3 and ^R4 together with the carbon atom to which they are attached form a methylene group;
A double line consisting of a solid line and a dashed line represents a single bond or a double bond.
The present invention relates to a method for producing a compound represented by the formula:
General Formula II:

(II)
(The symbols in the formula are the same as those in general formula I.)
The compound represented by the general formula III:

(The symbols in the formula are the same as those in general formula I.)
and converting the compound represented by the formula:
and converting said compound of general formula III into said compound of general formula I. In some cases, the substituents of said compound of general formula II and said compound of general formula III may be protected by any protecting group.

As a means for converting the compound represented by general formula II to the compound represented by general formula III, any means commonly used in the art can be adopted without particular limitation. For example, the compound represented by general formula II may be irradiated with ultraviolet laser light to cleave the bond at the 9-10 position of the steroid structure. The ultraviolet laser light is not particularly limited, but may have a wavelength of, for example, about 190 to 310 nm. The ultraviolet laser light may be oscillated by an oscillator commonly used in the art, for example, an argon ion laser, a krypton fluoride excimer laser, an argon fluoride excimer laser, a krypton chloride excimer laser, a xenon chloride excimer laser, a dye laser, a YAG laser, a YAG laser-pumped dye laser, an excimer laser-pumped dye laser, or a ruby laser.

As a means for converting the compound represented by general formula III to the compound represented by general formula I, any means commonly used in the art can be used without particular limitation. For example, the compound represented by general formula III may be isomerized by applying heat (e.g., heating at about 80 to about 120°C for about 1 to about 4 hours).

Each conversion step can be carried out in any solvent. The solvent is not particularly limited, but may be, for example, a hydrocarbon solvent such as hexane, heptane, cyclohexane, ligroin, benzene, toluene, or xylene; a halogenated hydrocarbon solvent such as bromobenzene, chlorobenzene, carbon tetrachloride, 1,2-dichloroethane, or 1,2-dibromoethane; an ether solvent such as diethyl ether, tetrahydrofuran, dioxane, or ethyl cellosolve; or an alcohol solvent such as methanol, ethanol, or propanol. In addition, the production method of the present invention may further include any step that is normally performed in the art, as long as it does not impair the object of the present invention.

The present invention will be described in detail below with reference to examples, but the scope of the present invention is not limited to these examples.

[Preparation Example]
(1) 988 mg (2.39 mmol) of fucosterol (compound 1) was dissolved in 7 mL of pyridine, 3.0 mL (31.7 mmol) of acetic anhydride was added, and the mixture was stirred at 45° C. for 3.5 hours. Ice was then added and the mixture was stirred for another hour. After the reaction was completed, the mixture was washed with 2N hydrochloric acid and extracted with chloroform. The solvent was removed with an evaporator, and the mixture was recrystallized from ethanol to obtain compound 2a (yield: 1061 mg, 97%). The results of optical rotation measurement, proton nuclear magnetic resonance measurement, and accurate mass analysis are shown below.

Compound 2a
[α] ²⁵ _D = -45.2 (c = 0.847, chloroform)
¹ H-NMR (400 MHz, deuterium chloroform): δ= 0.69 (s, 3H, Me-18), 0.92-1.32 (m, 8H, H-1a, H-9, H-12a, H-14, H-16a, H-17, H-22a, H-23a), 0.978 (d, 3H, J = 6.8 Hz, Me -26 or Me-27), 0.981 (d, 3H, J = 6.8 Hz, Me-26 or Me-27), 0.99 (d, 3H, J = 6.5 Hz, Me-21), 1.02 (s, 3H, Me-19), 1.37-1.64 (m, 8H, H-2a, H-7a, H-11a, H-1 1b, H-20, H-22b, H-23b, H-25), 1.57 (d, 1H, J = 6.6 Hz, Me-24 ² ), 1.82-1.91 (m, 4H, H-1b, H-2b, H-15a, H-16b), 1.94-2.10 (m, 3H, H-7b, H-12b, H-1 5b), 2.03 (s, 3H, Ac), 2.20 (m, 1H, H-8), 2.32 (m, 2H, H-4a, H-4b), 4.60 (m, 1H, H-3), 5.18 (q, 1H, J =6.7 Hz, H-24 ¹ ), 5.37 (d, 1H, J = 4.9 Hz , H-6); ¹³ C-NMR (100 MHz, CDCl ₃ ), δ = 11.8 (C-18), 13.2 (C-24 ² ), 18.8 (C-21), 19.3 (C-19), 21.0 (C-11), 21.4 (CO C H ₃ ), 22.1 (C-26 or C-27), 22.2 (C- 26 or C-27), 24.3 (C-23), 25.7 (C-15), 27.8 (C-2), 28.2 (C-16), 31.9 (C-7 or C-25), 31.9 (C-7 or C-25), 34.8 (C-8), 35.2 (C-22), 36.4 (C-20), 36.6 (C-10), 37.0 (C-1), 38.1 (C-4), 39.7 (C-12), 42.4 (C-13), 50.0 (C-9), 55.8 (C-17), 56.7 (C-14), 74.0 (C-3), 115.6 (C-24 ¹ ), 122.6 (C-6), 139.7 (C- 5), 147.0 (C-24), 170.5 (C=O)
APCI-Orbitrap-MS: calcd. for C ₃₁ H ₅₁ O ₂ ⁺ (M + H) ⁺ : 455.3884, found m/z: 455.3869.

(2) 300 mg (0.66 mmol) of compound 2a obtained in (1) was dissolved in 7.5 mL of cyclohexane at 65° C., 176 mg (0.989 mmol) of N-bromosuccinimide (NBS) was added, and the mixture was stirred at 100° C. for 1.5 hours under reflux conditions. The reaction temperature was lowered to room temperature, 15 mL of water was added, and the mixture was further stirred at room temperature for 1 hour. The reaction product was washed with water, extracted with normal hexane, and the solvent was distilled off under vacuum. 1.5 mL of 1M tetrabutylammonium fluoride-tetrahydrofuran (Bu ₄ NF-THF) solution was added to the obtained mixture, and the mixture was stirred at room temperature for 12 hours. The reaction product was washed with water, extracted with normal hexane, and concentrated. This was subjected to silica column chromatography, eluted with ethyl acetate/normal hexane (1:30), and separated and purified, and a fraction containing compound 2b (having a 5,7-diene structure) having a UV absorption region was obtained (184 mg).

(3) 184 mg of the fraction containing compound 2b obtained in (2) was dissolved in 2 mL of dichloromethane and 5 mL of methanol, and 28% sodium methylate (NaOMe) methanol solution was added until the pH reached 12, and the mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated and subjected to silica column chromatography, and eluted with ethyl acetate/normal hexane (3:7) to obtain fraction a (Figure 1) containing deacetylated compound 2c (104.31 mg).

(4) 95.31 mg of fraction a containing compound 2c obtained in (3) was dissolved in 25 mL of cyclohexane containing 0.1% 3-tert-butyl-4-hydroxyanisole (BHA) and transferred to a Petri dish. With stirring, the mixture was irradiated with 280 nm UV (9.71 mW/cm ² ) at room temperature for 2 hours. After irradiation, the reaction mixture was concentrated, subjected to silica column chromatography, and eluted with ethyl acetate/normal hexane (2:8) to obtain fraction b (FIG. 2) containing compound 2d (10.71 mg). This was dissolved in 10 mL of cyclohexane containing 0.1% BHA and stirred at 100° C. for 2 hours under reflux conditions. The reaction mixture was concentrated, subjected to silica column chromatography, and eluted with ethyl acetate/normal hexane (2:8) to obtain fraction c (FIG. 3) containing compounds 3 and 4. Fraction c was further subjected to high performance liquid chromatography (pump: Waters system, detector: UV detector, detection wavelength 215 nm, column: Kanto Chemical Mightysil RP-18 GP, mobile phase: acetonitrile, flow rate: 5 mL/min) to separate and purify the fractions eluting as two main peaks (peak A and peak B), thereby obtaining compound 3 (peak B, 4.10 mg) and compound 4 (peak A, 1.13 mg). The results of optical rotation measurement, proton nuclear magnetic resonance measurement, and accurate mass analysis are shown below.

Compound 3
[α] ²⁵ _D = +25.6 (c = 0.098, chloroform)
¹ H-NMR (400 MHz, deuterium chloroform), δ= 0.55 (s, 3H, Me-18), 0.979 (d, 3H, J = 6.7 Hz, Me-26 or Me-27), 0.983 (d, 3H, J = 6.8 Hz, Me-26 or Me-27), 0.99 (d, 3H, J = 5. 2 Hz, Me-21), 1.06 - 1.17 (m, 1H, H-22a), 1.26 - 1.42 (m, 5H, H-12a, H-16a, H-17, H-20, H-22b), 1.46 - 1.72 (m, 6H, H-2a, H-9a, H-11a, H-11b, H- 15a, H-15b), 1.57 (d, 3H, J = 6.9 Hz, Me-24 ² ), 1.83 - 2.11 (m, 6H, H-2b, H-12b, H-14, H-16b, H-23a, H-23b), 2.14 - 2.23 (m, 2H, H-1a, H-25), (dd, 1H, J = 13.3, 7.6 Hz, H-4a), 2.40 (m, 1H, H-1b), 2.57 (dd, 1H, J = 13.0, 3.8 Hz, H-4b), 2.82 (m, 1H, H-9b), 3.95 (m, 1H, H-3), 4.82 (d, 1 H, J= 2.4 Hz, H-19a), 5.05 (1H, H-19b), 5.19 (q, 1H, J = 6.7 Hz, H-24 ¹ ), 6.03 (d, 1H, J = 11.3 Hz, H-7), 6.24 (d, 1H, J = 11.2 Hz, H-6)
¹³ C-NMR (100 MHz, CDCl ₃ ), δ= 12.0 (C-18), 13.2 (C-24 ² ), 18.9 (C-21), 22.1 (C-15 or C-26 or C-27), 22.2 (C-15 or C-26 or C-27), 22.3 (C-15 or C-26 or C- 27), 23.6 (C-11), 25.7 (C-23), 27.7 (C-16), 29.0 (C-9), 31.9 (C-1), 34.8 (C-25), 35.2 (C-2 and C-22), 36.7 (C-20), 40.5 (C-12), 45.87 (C-4 or C-13), 45.92 (C-4 or C-13), 56.25 (C-14 or C-17), 56.33 (C-14 or C-17), 69.2 (C-3), 112.4 (C-19), 115.6 (C-24 ¹ ), 117.5 (C-7), 122.5 (C-6), 135.1 (C-5), 142.3 (C-8), 145.1 (C-10), 147.0 (C-24); NOESY-NMR (400 MHz, CDCl ₃ ), δ= 5.19 (H-24 ¹ ) → 0.979 or 0.983 (Me-26), 5.19 (H-24 ¹ ) → 1.57 (Me-24 ² ), 5. 19 (H-24 ¹ ) → 2.14 - 2.23 (H-25)
APCI-Orbitrap-MS, calcd. for C ₂₉ H ₄₇ O ⁺ (M + H) ⁺ : 411.3621, found m/z: 411.3620.

Compound 4
[α] ²⁵ _D = +8.9 (c = 0.023, chloroform)
¹ H-NMR (400 MHz, deuterium chloroform), δ= 0.55 (s, 3H, Me-18), 1.02 (d, 3H, J = 6.4 Hz, Me-21), 1.13 (m, 1H, H-22a), 1.23 - 1.35 (m, 3H, H-12a, H-16a, H-17), 1.38 - 1.59 (m, 5H, H-11a, H-15a, H-15b, H-20, H-22b), 1.64 - 1.72 (m, 3H, H-2a, H-9a, H-11b), 1.71 (d, 3H, J = 6.9 Hz, Me-24 ² ), 1.85 - 2.02 (m , 4H, H-2b, H-12b, H-14, H-16b), 1.87 (d, 1H, J = 0.8 Hz, Me-26), 2.08 - 2.21 (m, 2H, H-1a, H-23a), 2.26 - 2.43 (m, 3H, H-1b, H-4a, H-23b), 2.57 (dd, 1H, J = 13.0, 3.7 Hz, H-4b), 2.82 (m, 1H, H-9b), 3.95 (m, 1H, H-3), 4.82 (d, 1H, J = 2.5 Hz, H-19a), 4.86 (s, 1H, H-27a), 4.96 (s, 1H, H-27b) ), 5.05 (1H, H-19b), 5.63 (q, 1H, J = 6.9 Hz, H-24 ¹ ), 6.03 (d, 1H, J = 11.3 Hz, H-7), 6.24 (d, 1H, J = 11.3 Hz, H-6)
¹³ C-NMR (100 MHz, CDCl ₃ ), δ= 12.0 (C-18), 13.9 (C-24 ² ), 18.9 (C-21), 21.3 (C-26), 22.3 (C-15), 23.6 (C-11), 24.2 (C-23), 27.7 (C-16), 2 9.0 (C-9), 31.9 (C-1), 35.0 (C-2 or C-22), 35.2 (C-2 or C-22), 36.7 (C-20), 40.5 (C-12), 45.87 (C-4 or C-13), 45.92 (C-4 or C-13), 56.2 (C-14 or C-1) 7), 56.3 (C-14 or 143 .6 (C-25), 145.1 (C- ¹⁰ ); NOESY-NMR (400 MHz, CDCl 3 ), δ= 5.63 (H-24 1 ) → 1.71 (Me-24 ² ), 5.63 (H-24 ¹ ) → 1.87 (Me- ²⁶ ), 4.86 (H-27a) → _1.87 (Me-26), 4.96 (H-27b) → 2.08 - 2.21 (H-23a), 4.96 (H-27b) → 2.26 - 2.43 (H-23b)
APCI-Orbitrap-MS, calcd. for C ₂₉ H ₄₇ O ⁺ (M + H) ⁺ : 411.3621, found m/z: 411.3620.

化合物５ [Biological Activity]
Compounds 3 and 4 can be hydroxylated at the 25-position in the liver to give compound 5 below.

Compound 5

化合物６ Compound 5 can be hydroxylated at the 1-position in the kidney to produce the following compound 6. Compound 6 can act as active vitamin D in vivo.

Compound 6

化合物７

化合物８ Compounds 3 and 4 can be hydroxylated at the 1-position in the kidney to produce the following compounds 7 and 8, respectively. These compounds can also act as active vitamin D in vivo.

Compound 7

Compound 8

PASS (Prediction of Activity Spectra for Substances) online is widely used as a tool for predicting biological activity spectra based on the structural formula of a compound (http://www.way2drug.com/PASSOnline/index.php; see references below). PASS online is a tool for predicting the biological activity of a test compound based on the structure-activity relationship of known biologically active compounds. Since vitamin D derivatives with various structures are known, it is believed that in the case of compounds having a structure common to known vitamin D, such as the compounds of the present invention, it is possible to predict the biological activity with high reliability. Therefore, the chemical structures of compounds 6 to 8, which are the active forms of compounds 3 and 4, were analyzed by PASS online to obtain Pa values (the closer to 1, the higher the activity). The results of comparison with active vitamin D3 (VD3) and active vitamin D5 (VD5) are shown in Table 1.

As a result of the analysis using PASS Online, it was speculated that the compounds of the present invention have physiological effects common to vitamin D. Furthermore, their activity spectrum was distinctive. For example, although the Pa values of many of the compounds of the present invention were lower than that of vitamin D5, the Pa values of the apoptosis agonists were higher than that of vitamin D5. Compiling compounds with such distinctive activity spectra is useful not only for the search for vitamin D derivative compounds, but also for their use as functional materials.

In general, active vitamin D has the ability to induce cell differentiation and is expected to be effective as an anticancer drug (Rao D.S. et al., Steroids, 2001, 66, 423-431), but active vitamin D3 has a strong calcium absorption stimulating effect, so if the active form is directly ingested, side effects such as hypercalcemia and increased calcification of soft tissues may occur. Therefore, active vitamin D3 is difficult to use as an anticancer drug. In contrast, 1-OH-vitamin D5 has a lower calcium absorption stimulating effect than active vitamin D3, so it has been attempted to be used as an anticancer drug (Lazzaro G., et al., Eur. J. Cancer, 2000, 36, 780-786 and Guyton K.Z. et al., Nutr. Rev., 2003, 61, 227-238). 1-OH-vitamin D5 does not have the side effects of active vitamin D3 and has been shown to be effective against breast cancer, colon cancer, and prostate cancer. The compound of the present invention also has a low Pa value for calcium regulation, so it may be used as an anticancer drug with a low risk of side effects such as hypercalcemia and increased calcification of soft tissues.

*References for PASS
(a) Komba S., et al., J. Oleo Sci., 2015, 64(9), 1009-1018
(b) Lagunin, A., et al., Bioinformatics, 2000, 16, 747-748
(c) Poroikov, V., et al., Sar Qsar Environ. Res., 2001, 12, 327-344
(d) Stepanchikova, A., et al., Curr. Med. Chem., 2003, 10, 225-233
(e) Lagunin, A., et al., Curr. Pharm. Des. 2010, 16, 1703-1717
(f) Filimonov, D., et al., Chem. Heterocycl. Compd., 2014, 50, 444-457

As a result, we were able to produce a new vitamin D derivative compound. This has expanded the variety of available vitamin D derivative compounds, increasing the options for vitamin D derivative compounds that can be used for different purposes.

Claims (12)

General Formula I:

(I)
(Wherein,
^R1 is an OH group, an O— _C2 - _C18 acyl group, or a glycoside group;
^R2 is H, an OH group, or an O- _C2 - _C18 acyl group;
^R3 and ^R4 are each independently H, a methyl group, or an OH group, or ^R3 and ^R4 together with the carbon atom to which they are attached form a methylene group;
A double line consisting of a solid line and a dashed line represents a single bond or a double bond.
A compound represented by the formula: The compound or stereoisomer thereof according to claim 1, wherein the double line consisting of a solid line and a dashed line represents a single bond. 3. The compound or stereoisomer thereof according to claim 1 or 2, wherein R ¹ is an OH group and R ² is H. 3. The compound or stereoisomer thereof according to claim 1 or 2, wherein R ¹ is an OH group and R ² is an OH group. The compound or stereoisomer thereof according to any one of claims 1 to 4, wherein R ³ is a methyl group and R ⁴ is H, or R ³ and R ⁴ together with the carbon atom to which they are attached form a methylene group. The compound or stereoisomer thereof according to any one of claims 1 to 4, wherein R ³ is a methyl group and R ⁴ is OH. R ¹ is an OH group,
R ² is an H or OH group,
R ³ is a methyl group, and R ⁴ is H or OH, or R ³ and R ⁴ form a methylene group together with the carbon atom to which they are attached,
The double line consisting of a solid line and a dashed line represents a single bond.
The compound according to claim 1 or a stereoisomer thereof. R ¹ is an OH group,
R ² is H,
R ³ is a methyl group, and R ⁴ is H or R ³ and R ⁴ form a methylene group together with the carbon atom to which they are attached,
The double line consisting of a solid line and a dashed line represents a single bond.
The compound according to claim 1 or a stereoisomer thereof. The compound or a stereoisomer thereof has the following formula:

2. The compound of claim 1, or a stereoisomer thereof, selected from the group consisting of: The compound or a stereoisomer thereof has the following formula:

2. The compound of claim 1, or a stereoisomer thereof, selected from the group consisting of : A composition comprising the compound according to any one of claims 1 to 10 or a stereoisomer thereof. General Formula I:

(I)
(Wherein,
^R1 is an OH group, an O— _C2 - _C18 acyl group, or a glycoside group;
^R2 is H, an OH group, or an O- _C2 - _C18 acyl group;
^R3 and ^R4 are each independently H, a methyl group, or an OH group, or ^R3 and ^R4 together with the carbon atom to which they are attached form a methylene group;
A double line consisting of a solid line and a dashed line represents a single bond or a double bond.
A method for producing a compound represented by the formula:
General Formula II:

(II)
(The symbols in the formula are the same as those in general formula I.)
The compound represented by the general formula III:

(The symbols in the formula are the same as those in general formula I.)
and converting the compound represented by the formula:
converting said compound of general formula III to said compound of general formula I;
A manufacturing method comprising: