JPS6231702B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a method for producing active vitamin D ₃ . More specifically, the present invention provides an industrially extremely valuable method for producing active vitamin D ₃ useful for regulating calcium metabolism in warm-blooded animals in high yield. Conventionally, for example, the method for producing 1α-hydroxycholecalciferol, which is one of the active forms of vitamin _D3 , has been described in JP-A-48-62750, which involves 17 or more steps starting from cholesterol. A method for producing 1α-hydroxycholecalciferol is known, and JP-A-51-110554 also describes a completely different method for producing 1α-hydroxycholecalciferol starting from cholesterol. It has been known. In both of these methods, 1α,3β-diacetoxycholesterol
By irradiating 5,7-diene with ultraviolet light, 1α,3β-
The final step is described as diacetoxyprevitamin D ₃ and then converting it to 1α,3β-diacetoxyvitamin D ₃ . However, for example, as described in the example of JP-A-48-62750, 1α,3β-diacetoxycholester-5,7-diene produced by irradiating 600 μg of 1α,3β-diacetoxycholester-5,7-diene with ultraviolet light is The yield of acetoxy previtamin D ₃ is 120 μg (yield 20%)
Moreover, Example 3 of JP-A No. 51-110554
As described in 135 mg of 1α,3β-diacetoxycholester-5,7-diene, 13 mg of 1α-hydroxycholecalciferol and 8 mg of 1α-hydroxy previtamin D ₃ were obtained. do not have. Based on the patent applications of the pioneers regarding the manufacturing method of these two 1α-hydroxycholecalciferols, as well as other patent applications and literature publications published after that, it is clear that so-called active vitamins such as 1α-hydroxycholecalciferol As a method for producing D ₃ , the reaction process of producing previtamin D ₃ by irradiating the corresponding 1α,3β-dihydroxycholester-5,7-dienes or their hydroxyl-protected derivatives with ultraviolet rays is an industrial method. It is considered to have extremely important meaning as a manufacturing process for active vitamin _D3 . However, as is clear from the above example,
The total number of steps to produce active vitamin D ₃ is extremely large, and therefore the overall yield naturally decreases. However, the yield improvement in the ultraviolet irradiation step, which is close to the final step, is due to the direct target product, the active form. vitamin
It has extremely important industrial significance because it contributes to improving the yield of D ₃ . From this point of view, the present inventors have determined that 1α,3β-
As a result of researching various reaction conditions in the process of producing previtamin D ₃ by irradiating dihydroxycholester-5,7-dienes with ultraviolet light, we have discovered a large number of new facts. Previously, for example, results have been reported that clarified the decrease in ergosterol with respect to the irradiation time when ergosterol is irradiated with ultraviolet rays, and the behavior of the generated previtamin D ₂ and the corresponding tachysterol. It is known that the production of previtamin D ₂ reaches about 25% saturation after about 2 hours of irradiation, and that the production of the corresponding tachysterol increases with the irradiation time (``vitamin'' 47 5 Issue (May) 187~
200 (1973)). However, according to the research of the present inventor, 1
In the case of α,3β-dihydroxycholester-5,7-dienes, it is certainly the case that the corresponding previtamins
D ₃ and tachysterol are also produced, but the reaction is extremely complex, with various other by-products produced, and therefore, for example, in order to increase the production of previtamin D ₃ -Increasing the conversion of dihydroxycholester-5,7-dienes not only increases the production of taxerol, but also increases the production of previtamin _D3 or vitamin D3.
Furthermore, various by-products that are difficult to separate from D ₃ accumulate, which not only does not significantly improve the yield of the target active vitamin D ₃ , but also tends to become impure, which is undesirable. It became clear. According to the method of the present invention, 1α,3β-
Under optimal conditions, active vitamin D ₃ can be produced from dihydroxycholester-5,7-dienes at a high yield of 80 to 90% or more. That is, the present invention provides the following formula [-a] which is a precursor of active vitamin D ₃ [In the formula, R ¹ and R ² are the same or different and are a hydrogen atom or a protecting group for a hydroxyl group, and R is a 6-methylheptyl-2-yl group or a hydroxyl group, a protected hydroxyl group, an oxo group, a protected oxo group , the corresponding group substituted by a halogen atom. ] 1α,3β-dihydroxycholester-5,7-dienes represented by are irradiated with ultraviolet rays to form the following formula [-a] [In the formula, the definitions of R ¹ , R ² and R are the same as above. ] The following formula [-a] is produced by converting to previtamin D ₃ represented by 3 and then isomerizing the previtamin D ₃ using thermal energy. [In the formula, the definitions of R ¹ , R ² and R are the same as above. ] In the method for producing active vitamin D type ₃ represented by the formula [- Previtamin D ₃ of formula [-a] and/or active vitamin D ₃ of formula [-a] are reacted until the amount becomes 0.03 to 0.4 parts by weight, and then from the reaction mixture, formula [-a]
Previtamin D type ₃ and/or formula [-
While separating the active vitamin D type ₃ of a], the unreacted 1α,3β-dihydroxycholester-5,7-dienes of formula [-a] are recycled and used as a precursor for the ultraviolet irradiation. This is a method for producing active vitamin D type ₃ , which is characterized by the following. The method of the present invention has two main steps as described above. One is active vitamin D ₃
The first step is to irradiate ultraviolet rays to 1α,3β-dihydroxycholester-5,7-dienes, which are precursors of 1α,3β-dihydroxycholester-5,7-dienes. The more active forms of vitamin D ₃ and/or their corresponding previtamin D ₃ are separated, and unreacted 1α, 3
This is a step in which β-dihydroxycholester-5,7-dienes are circulated as a precursor for the first step. According to the first step, the reaction itself is already known, and the 1α,3β-dihydroxycholest-5,7-dienes of the formula [-a] are irradiated with ultraviolet rays, and the single bonds at positions 9 and 10 are is cleaved and the corresponding [-a] previtamin D ₃ is produced. As mentioned above, the active vitamin D ₃ precursor used as a raw material in the method of the present invention is a compound having a cholesterol skeleton having a hydroxyl group or a protected hydroxyl group at the 1α-position, but especially the following formula [] [In the formula, R ¹ and R ² are the same or different and are a hydrogen atom or a hydroxyl group-protecting group. R ³ , R ⁴ and R ⁵ are the same or different and are a hydrogen atom, a hydroxyl group or a protected hydroxyl group, and R ³ and R ⁴ may be taken together to form a carbonyl group or a protected carbonyl group. A compound represented by is preferably used. In the above formula [], the hydroxyl protecting group representing R ¹ , R ² , R ³ , R ⁴ and R ⁵ is the active vitamin represented by the above formula [-a] or the below-mentioned formula [].
D It is a protecting group that can be converted into a hydroxyl group without changing the structure of Group ₃ , and examples include the following groups. (1) Acyl group; For example, acetyl group, propanoyl group, butanoyl group, pentanoyl group, pivaloyl group, capronyl group, cyclohexanoyl group, chloroacetyl group, bromoacetyl group, benzoyl group, p
C1-C12 aliphatic or aromatic carboxylic acid residues such as -bromobenzoyl group, p-nitrobenzoyl group, ethylbenzoyl group, toluyl group, or their nitro-, halogen-, or alkoxy-substituted derivatives are preferably used. Among these, particularly preferred are acetyl group, benzoyl group, propanoyl group, and the like. (2) A group that forms an ether bond with a hydroxyl group; For example, a t-butyl group, a benzyl group, a triallylmethyl group such as a triphenylmethyl group, an alkyl-substituted silyl group such as a tetrahydropyranyl group, a methoxymethyl group, a trimethylsilyl group, etc. etc. can be given. Among the above protecting groups, the acyl group (1) is particularly preferably used, but the present invention is not particularly limited thereto. Specific examples of the 1α,3β-dihydroxycholester-5,7-dienes represented by the above formula [-a] or the above formula [] include, for example, 1α,3β-dihydroxycholester-5,7-dienes.
-diene, 1α,3β-25-trihydroxycholesta-5,7-diene, 1α,3β,24-trihydroxycholesta-5,7-diene, 1α,3
β,24(S)-trihydroxycholester-5,7
-diene, 1α,3β,24(R)-trihydroxycholesta-5,7-diene,1α,3β,24,
25-tetrahydroxycholesta-5,7-diene, 1α,3β-dihydroxy-24-oxocholesta-5,7-diene, 24(24)-ethylenedioxy-1α,3β-dihydroxycholesta-5,
7-diene and, for example, 1α,3β-diacetoxycholester-5,7-diene, 1α,3β-dipivaloyloxycholester 5,7-diene, 1
Hydroxyl-protected derivatives of 1α,3β-dihydroxycholester-5,7-diene such as α,3β-di(trimethylsilyloxy)cholester 5,7-diene can be mentioned. Of course, other 1α,3β-dihydroxycholesterols such as those mentioned above
Corresponding protected derivatives of 5,7-dienes as mentioned above are also exemplified, but since specific examples are obvious in themselves, no individual examples will be given. The first step is the reaction as described above, but the reaction is carried out in an inert organic solvent by irradiating it with ultraviolet light. The ultraviolet light used at this time is approximately 200 to 360 nm.
It is known to have a wavelength range of 260 to 310 nm, and in the method of the present invention, a wavelength range of 260 to 310 nm is particularly preferably used. In addition, examples of inert organic solvents include hexane, heptane, cyclohexane, ligroin,
Hydrocarbons such as benzene, toluene, xylene, bromobenzene, chlorobenzene, nitrobenzene, carbon tetrachloride, 1,2-dichloroethane, 1,2-dibromoethane, halogenated hydrocarbons, and diethyl ether, tetrahydrofuran, dioxane, methyl Ether solvents such as cellosolve, phenyl cellosolve, methanol, ethanol,
Alcohol solvents such as propanol, hexanol, and cyclohexanol are often used as suitable solvents. In particular, single or mixed solvents such as benzene, toluene, diethyl ether, methanol, and ethanol are preferred, and when such solvents are used, the isomerization described below can be performed in the same solvent after ultraviolet irradiation. The temperature during UV irradiation is -20℃ to 80℃, especially -
A range of 10°C to 20°C is preferred. Further, it is preferable to carry out the reaction in an inert atmosphere in which oxygen does not exist, such as an argon or nitrogen atmosphere. Thus, according to ultraviolet irradiation, the above formula [−
The single bonds at the 9 and 10 positions of the 1α,3β-dihydroxycholesta-5,7-diene represented by [a] are cleaved to produce previtamin D ₃ represented by the above formula [-a]. The generated previtamin D ₃ is isomerized by thermal energy and converted into the corresponding active vitamin D ₃ . This isomerization reaction due to thermal energy is a reaction with an equilibrium value and rate that depend on temperature, and the lower the temperature, the lower the amount of previtamin D ₃
It shows an equilibrium value in which the proportion of the class increases, and on the other hand,
The higher the temperature, the faster the conversion rate from pre-vitamin D ₃ to active vitamin D ₃ . Therefore, the pre-vitamin D ₃ produced by ultraviolet irradiation as described above is already gradually isomerized into active vitamin D ₃ during the ultraviolet irradiation process. When primarily producing pre-vitamin D ₃ , it is desirable that the ultraviolet irradiation in this first step be performed at a low temperature and for a short period of time, where the isomerization rate is _low . In this case, the UV irradiation temperature and time should be considered as well, and after UV irradiation, the temperature should be maintained at a suitable temperature for a certain period of time (e.g., 2 to 3 seconds at 80℃).
It is desirable to actively carry out isomerization depending on the amount of time (time, etc.), and then carry out the second separation step. The greatest feature of the method of the present invention is that the ultraviolet irradiation reaction in this first step
D ₃ and/or the amount of active vitamin D ₃ produced when these are isomerized by thermal energy is determined by the amount of unreacted 1α,3β-dihydroxycholester-5, The ultraviolet irradiation should be continued until the amount reaches 0.03 to 0.4 parts by weight, more preferably 0.15 to 0.35 parts by weight, and should not be continued until the amount reaches 0.15 to 0.35 parts by weight, per 1 part by weight of the 7-diene. According to the research of the present inventor, active vitamin _D3
In the reaction of irradiating ultraviolet light to 1α,3β-dihydroxycholester-5,7-dienes, which are the precursors of
Not only tachysterol, but also at least two other by-products are produced, and the amount of these products increases as the consumption of 5,7-dienes, which are raw materials, increases. Sufficient facts were observed to infer that it was converted into a product. Therefore, the produced previtamin D ₃ and/or
Alternatively, if the amount of active vitamin D ₃ produced from this exceeds the above upper limit relative to unreacted 1α,3β-dihydroxycholester-5,7-dienes, the reaction in the second step It is difficult to separate previtamin D ₃ and/or active vitamin D ₃ from the mixture with good purity, and unreacted 1α,3β-dihydroxycholesta-5,7-dienes are collected and used. However, if these products are recycled as they are, many by-products and their amounts will gradually accumulate in the reaction system.
In the end, it became clear that an industrial method for reusing unreacted 1α,3β-dihydroxycholesta-5,7-dienes could not be achieved.
Of course, the lower limit value of 0.03 for the production amount is determined because a lower value would result in too large a number of circulations and an undesirable amount of circulation. Thus, after passing through the first step, the reaction mixture is separated in the second step, and the unreacted 1α,3β-dihydroxycholester-5,7-dienes are used as a precursor raw material for the first step. be done. As mentioned above, the reaction mixture obtained in the first step contains previtamin D ₃ and/or active vitamin D ₃ . It was also mentioned above that when attempting to produce previtamin D ₃ , it is desirable to carry out the first step at a relatively low temperature and in a short period of time. However, active vitamin
When attempting to produce Type D ₃ , the reaction mixture obtained in the first step may be held until the isomerization reaction has sufficiently proceeded, and then subjected to the second step. Furthermore, the previtamin D ₃ separated in the second step may be subjected to an isomerization reaction using thermal energy. As mentioned above, the equilibrium value and rate of conversion of previtamin D ₃ to active vitamin D ₃ differ depending on temperature. Therefore, taking into account the equilibrium value and the conversion rate, the temperature can be determined, and in this sense the temperature is not essentially important for the progress of the reaction itself. Practically, taking these points into consideration, the isomerization temperature is usually 10 to 120°C, preferably 40 to 100°C. In this case, the isomerization is usually preferably carried out in an inert organic solvent, and if the above-mentioned suitable solvents are used, the same solvent as used in the ultraviolet irradiation can be used. Thus, according to the isomerization reaction by thermal energy, from the previtamin D ₃ represented by the above formula [-a], the following formula [-a] [In the formula, . The definitions of R ¹ , R ² and R are the same as above. ] and the following formula [] [In the formula, the definitions of R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are the same as above. ] _Three active forms of vitamin D are formed. According to the research of the present invention, when performing the second step, the above reaction mixture subjected to separation is
Of course, 0.03 parts by weight of 1α-hydroxycholester-5,7-dienes present in the reaction system.
~0.4 parts by weight of pre-vitamin D ₃ and active vitamin D ₃ , but if it is primarily active vitamin D ₃ , it is probably a combination of pre-vitamin D ₃ and active vitamin D ₃ . Although this seems to be due to an essential difference in stability, it has become clear that separation can be carried out very easily. The separation of previtamin D ₃ and/or active vitamin D ₃ from the reaction mixture in the second step is carried out by column chromatography, preparative thin layer chromatography, high performance liquid chromatography, recrystallization method, etc. You can do it by leaning. It is also possible to use chromatography using a carrier containing silicon dioxide (SiO ₂ ) adsorbing nonmetallic silver, such as silica gel containing silver nitrate. Furthermore, by combining two or more of these separation and purification methods, it is possible to produce even higher purity previtamin D ₃ and/or active vitamin D ₃ . According to the research of the present inventor, among the above separation methods, the separation method using high performance liquid chromatography, especially the separation method using high performance liquid chromatography using silica gel as the main component of the carrier, has a high separation yield. It has become clear that this is extremely preferable in terms of particularly increasing the . When a hydroxyl-protected derivative of previtamin D ₃ or an active vitamin D ₃ is obtained by the above separation, when the protecting group is an acyl group, it is usually treated with an alkaline alcohol such as methanol or ethanol. This is carried out by a method of decomposition in a solution or a method of deacylation by reductive decomposition with LiAlH ₄ or the like in a solvent such as ether. The deacylation reaction is preferably carried out at a temperature of -10°C to 50°C. Further, when the protecting group has an ether bond with the hydroxy group, a part of it can be easily removed by reductive removal or by contacting with an acid or an alkali. Thus, the separation means used in the second step can be used as is to purify the previtamin D ₃ or active vitamin D ₃ obtained by the deprotection reaction. As detailed above, according to the method of the present invention, 1α, 3
From β-dihydroxycholester-5,7-dienes, it is possible to produce active vitamin D ₃ , which is effective for regulating calcium metabolism in warm-blooded animals. Hereinafter, the present invention will be explained in more detail with reference to Examples. Example 1 (a) First reaction: 1α,3β-diacetoxycholester-5,7-
Dissolve 2.660 g of diene in 3.3 anhydrous benzene, and add a 200W high-pressure mercury lamp (manufactured by Hanovia) to the solution.
654-A), and was irradiated with ultraviolet rays for 4 minutes in an argon gas atmosphere through a Vycor filter. The obtained ultraviolet irradiation mixture was heated to No. 25-4.
shall be. A portion of this is sampled and subjected to liquid chromatography [HLC]. Column: Silica gel (Zorbax SIL) 25cm x 2.1mmφ eluent;
CH ₂ Cl ₂ /n-hexane = 1/3, detection; 254 mμ
(UV-254 detector)]. The results are shown in Table-1. Next, this No. 25-4
was divided into two equal parts. One of them is No.25−4U
shall be. The other side was refluxed at 80°C for 2 hours in an argon atmosphere. This thermal isomerization mixture is designated as No. 25-4H. A portion of this No. 25-4H was sampled and subjected to HCL analysis under the same conditions as above. The results are shown in Table-1. No. 25-4H and No. 25-4U were separately heated at 30°C under reduced pressure.
It was concentrated to dryness as follows. The total amount of each concentrated dry product was separated by chromatography using a column packed with 80 g of silica gel. The developing solvent used was sequentially changed to benzene-hexane, benzene, and then benzene-ethyl acetate. The results are shown in Table-2. (b) Second reaction: 1α, 3 obtained by column chromatography from the above No. 25-4U and No. 25-4H, respectively.
The total amount of the fraction containing β-diacetoxycholester-5,7-diene was treated separately under the same conditions as in the first reaction above, except that the UV irradiation time was 7 hours.
Ultraviolet irradiation was performed for a few minutes. Among the obtained ultraviolet irradiation mixtures, the one obtained from No. 25-4U is designated as No. 26-7U. On the other hand, the ultraviolet irradiated product obtained from No. 25-4H was thermally isomerized under the same conditions as the first reaction. This thermal isomerization mixture is designated as No. 26-7H. A portion of these two No. 26-7U and No. 26-7H was taken and subjected to HLC analysis in the same manner as above, and the total amount of each was separated by column chromatography in the same manner as above. . The results are shown in Table-1 and Table-2, respectively. (c) 3rd to 5th reactions: 1α and 3β obtained by column chromatography from the above Nos. 26-7U and 26-7H, respectively.
-Diacetoxycholester-5,7-diene-containing fractions were separately subjected to repeated reactions in the same manner as in the second reaction. The fourth and fifth reactions were repeated in the same manner. However, the concentrations during the third to fifth UV irradiations are different from those during the first and second times,
The concentration was reduced by 25%. In addition, the ultraviolet irradiation time was 5 minutes for both the third and fourth times, and the time for the fifth time was 3 minutes in consideration of the ultraviolet irradiation efficiency because the raw material recovered from the fourth time was reduced. The results are shown in Table-1 and Table-2. In addition, in the table, No. 27-5U means an ultraviolet irradiation mixture using the raw material recovered from No. 26-7U, and No. 27-5H, No. 28-5H, and No. 29- 3H means a thermal isomerization mixture using raw materials recovered from No. 26-7H, No. 27-5H, and No. 28-5H, respectively. As is clear from the results in Table 1, according to the method of the present invention, 1α,3β-diacetoxy cholesta 5,7
-1α,3β-diacetoxyvitamin from diene
It can be seen that D ₃ and 1α,3β-diacetoxy previtamin D ₃ can be produced.

[Table] In the table, 2p, 10p, and 12p, which represent byproducts, are
It is confirmed as a clear peak by HLC, and 2p is detected earlier than the target substance and raw material, and 10p and 12p are both detected later than the target substance and raw material. be. Further, the conversion rate is the consumption rate (%) of the raw material, and the selectivity is the production rate (%) of the target product with respect to the consumed raw material. Moreover, the average values of conversion rate and selectivity in the upper row are the average values for the U system, and the lower rows are the average values for the H system.

【table】

[Table] Example 2 In the same manner as in Example 1, the circulation reaction (ultraviolet irradiation and thermal isomerization) was repeated five times using 1α,3β-dihydroxycholesta-5,7-diene as the starting material. The thermal isomerization mixture of each time was prepared in the same manner as in Example 1.
HLC analysis was performed (however, CH ₂ Cl ₂ containing 3% methanol was used as the eluent), and the average conversion was 21.3%.
(The average weight ratio of the target product to the unreacted raw material was 0.23), and the average selectivity was 86.2%. Example 3 In the same manner as in Example 1, the cyclic reaction was repeated three times using 1α,3β-dipivaloyloxycholester-5,7-diene as the starting material. The thermal isomerization mixture of each round was subjected to HLC analysis in the same manner as in Example 1 (however, the gradient method was used using isooctane containing 2 to 20% isopropyl alcohol as the eluent), and the average conversion rate was 23.6% (produced target product). Average weight ratio to unreacted raw material: 0.26), average selectivity
Obtained a result of 92.5%. Example 4 In the same manner as in Example 1, the cyclic reaction was repeated three times using 1α,3β,24-trihydroxycholester-5,7-diene as the starting material. The thermal isomerization mixture of each round was subjected to HLC analysis in the same manner as in Example 1 (however, the gradient method was used using n-hexane containing 1 to 10% isopropyl alcohol as the eluent), and the average conversion rate was 22.7%. Average weight ratio of target product to unreacted raw material: 0.21), average selectivity
Obtained a result of 84.7%.

Claims (1)

[Claims] 1. The following formula [-a] which is a precursor of active vitamin D ₃ [In the formula, R ¹ and R ² are the same or different and are a hydrogen atom or a protecting group for a hydroxyl group, and R is a 6-methylheptyl-2-yl group, a hydroxyl group, a protected hydroxyl group, an oxo group, a protected oxo group , the corresponding group substituted by a halogen atom. ] 1α,3β-dihydroxycholester-5,7-dienes represented by are irradiated with ultraviolet rays to form the following formula [-a] [In the formula, the definitions of R ¹ , R ² and R are the same as above. ] The following formula [-a] is produced by converting to previtamin D ₃ represented by 3, and then isomerizing the previtamin D ₃ with thermal energy. [In the formula, the definitions of R ¹ , R ² and R are the same as above. ] In the method for producing active vitamin D type ₃ represented by [
-a] of 1α,3β-dihydroxycholesta-
0.03 to 0.4 parts by weight of previtamin D ₃ of formula [-a] and/or active vitamin D ₃ of formula [-a] generated in the reaction system per 1 part by weight of 5,7-dienes. The reaction mixture was then reacted until the previtamin of formula [-a]
D ₃ and/or active vitamin D ₃ of formula [-a] are separated, and unreacted formula [-a] is separated.
-a] of 1α,3β-dihydroxycholesta-
A method for producing active vitamin D ₃ , characterized in that 5,7-dienes are recycled as a precursor for the ultraviolet irradiation. 2. Active vitamin D ₃ according to claim 1, wherein previtamins and/or active vitamin D ₃ are separated from the reaction mixture by high performance liquid chromatography using silica gel as a carrier. manufacturing method. 3 1α,3β-dihydroxycholesta-5,7
−The dienes have the following formula [] [In the formula, R ¹ and R ² are the same or different and are a hydrogen atom or a hydroxyl group-protecting group. R ³ , R ⁴ and R ⁵ are the same or different and are a hydrogen atom, a hydroxyl group or a protected hydroxyl group, and R ³ and R ⁴ may be taken together to form a carbonyl group or a protected carbonyl group. ] A method for producing active vitamin D ₃ according to claim 1 or 2, which is a compound represented by: 4 1α,3β-dihydroxycholesta-5,7
- The dienes are 1α,3β-dihydroxycholester-5,7-diene, 1α,3β,25-trihydroxycholester-5,7-diene, 1α,3β,
24-trihydroxycholester-5,7-diene,
1α,3β,24,25-tetrahydroxycholester-5,7-diene, 1α,3β-dihydroxy-
The active vitamin D ₃ according to any one of claims 1 to 3, which is any compound selected from the group consisting of 24-oxocholesta-5,7-diene and hydroxyl-protected derivatives thereof. manufacturing method of types.