JP-7857399-B2 - Steroid compounds, methods for preparing them, and their use

Inventors

• 于 垂亮
• 李 小林
• 王 延東
• 蘭 小兵
• ▲はお▼ 飛
• 蘇 映雪
• 賀 海鷹
• 呉 美容

Assignees

• コアンチョウ オキュサン オフサルミック バイオテクノロジー カンパニー リミテッド
• オキュサン・オフサルミック・ファーマスーティカル（コアンチョウ）・カンパニー・リミテッド

Dates

Publication Date

20260512

Application Date

20220830

Priority Date

20210831

Claims (9)

1. A compound having the structure represented by formula I. (Here, X is triethylsilyl.)
2. Compounds having the structure represented by formula I and compounds having the structure represented by formula I' are (Here, X is triethylsilyl) A method for separating and purifying a compound of formula I according to claim 1, characterized by comprising the step of separating and purifying a raw material A containing a compound of formula I and a compound of formula I' by column chromatography to obtain a compound of formula I.
3. A method for separating and purifying a compound of formula I according to claim 2, further comprising the step of reacting crude lanosterol with a hydroxyl protective agent to obtain the raw material A, wherein the hydroxyl protective agent is triethylchlorosilane.
4. A method for purifying a compound of formula I, comprising the step of obtaining the compound of formula I by recrystallizing a raw material A containing the compound of formula I and the compound of formula I'. (Here, the definition of X is as described in claim 1, and the solvent for recrystallization is selected from 1) a mixture of ethyl acetate and isopropyl alcohol, or 2) isopropyl acetate.)
5. A purification method according to claim 4, wherein any one of the following conditions is met. (1) The mass percentage of the compound of formula I in raw material A is 90% or more. (2) In a mixture of ethyl acetate and isopropyl alcohol, the volume ratio of ethyl acetate to isopropyl alcohol is 1:(0.5 to 2).
6. 1) A step of reacting crude lanosterol with a hydroxyl protective agent to obtain a starting material A containing the compound of formula I and the compound of formula I' described in claim 2; The process includes: 2) separating the raw material A by column chromatography to obtain the compound of formula I; and 3) hydroxy-deprotecting the compound of formula I to obtain purified lanosterol; Here, the crude lanosterol comprises lanosterol and dihydrolanosterol; The hydroxyprotective agent is triethylchlorosilane. A method for purifying lanosterol characterized by the following features.
7. The separation and purification method according to claim 3, wherein the reaction between the crude lanosterol and the hydroxyprotecting agent satisfies any one of the following conditions. (1) The reaction between crude lanosterol and a hydroxyl protective agent is carried out in the presence of an organic solvent, the organic solvent being N,N-dimethylformamide or dichloromethane. (2) The reaction between crude lanosterol and a hydroxyprotectant is carried out in the presence of an acid binder, the acid binder being pyridine, imidazole, diisopropylamine, triethylamine, triethanolamine, potassium carbonate, or sodium carbonate. (3) The reaction between crude lanosterol and the hydroxyprotectant is carried out at a temperature of 60–85°C.
8. The method according to claim 6, wherein the reaction between the crude lanosterol and the hydroxyprotectant satisfies any one of the following conditions. (1) The reaction between crude lanosterol and a hydroxyl protective agent is carried out in the presence of an organic solvent, the organic solvent being N,N-dimethylformamide or dichloromethane. (2) The reaction between crude lanosterol and a hydroxyprotectant is carried out in the presence of an acid binder, the acid binder being pyridine, imidazole, diisopropylamine, triethylamine, triethanolamine, potassium carbonate, or sodium carbonate. (3) The reaction between crude lanosterol and the hydroxyprotectant is carried out at a temperature of 60–85°C.
9. The method according to claim 6, wherein the column chromatography is silica gel column chromatography, and the silica gel column chromatography satisfies any one of the following conditions. (1) The specifications of the silica gel selected for the silica gel column chromatography are 100-200 mesh, 200-300 mesh, or 300-400 mesh. (2) The eluent used in the silica gel column chromatography is one or more of petroleum ether, n-heptane, n-hexane, dichloromethane, and ethyl acetate. (3) Ammonia water is added to the eluent used in the silica gel column chromatography.

Description

This application claims priority to Chinese Patent Application No. 2021110137424, filed on August 31, 2021. This application incorporates the entire text of the aforementioned Chinese Patent Application. This invention relates to the field of separation and purification of organic compounds, and more specifically to steroid compounds, methods for preparing them, and their use. Lanosterol belongs to the tetracyclic triterpenoid group and is an intermediate in cholesterol biosynthesis. Currently, lanosterol is mainly obtained by isolation and extraction from crude lanosterol. However, crude lanosterol is a mixture of tetracyclic triterpenoids separated from lanolin by crystallization, and generally contains about 60% lanosterol (CAS number: 79-63-0), about 30% dihydromanosterol (CAS number: 79-62-9), and other impurities such as cholesterol (CAS number: 57-88-5). Of these, lanosterol and dihydrolanosterol are very similar in polarity, making their separation extremely difficult. CN101691391 reports the preparation of lanosterol with a separation purity of 97% using high-performance liquid chromatography, but the amount prepared is only 250 mg, making it costly and difficult to produce commercially on a large scale. Currently, there are no purification processes for the large-scale production and preparation of lanosterol with a purity exceeding 90%, making it difficult to meet the needs of medical applications. Furthermore, separating lanosterol from other impurity compounds is extremely difficult, and completely separating lanosterol by column chromatography or recrystallization is virtually impossible. For example, one study found that after recrystallizing lanosterol mixed with other impurities, the proportion of impurities in the precipitated lanosterol crystals did not change significantly from that before crystallization. Therefore, in order to better study lanosterol and utilize it in the medical field, there is an urgent need in the market for methods to isolate and purify lanosterol. This invention provides a steroid compound, a method for preparing the same, and its use to solve one of the technical problems in the prior art. Furthermore, this invention provides a method for purifying lanosterol that can separate and purify lanosterol from crude lanosterol. This method is simple to operate, has a stable process, high production capacity, low cost, and produces lanosterol with high purity suitable for various pharmaceutical applications. One aspect of the present invention provides a compound having a structure represented by formula I. According to some embodiments of the present invention, X is TMS, TES, TBS, TBDPS, TIPS, DMIPS, TBDMS, or MDIPS. Another aspect of the present invention provides a method for preparing a compound of formula I, comprising the step of reacting lanosterol with a hydroxyprotective agent to obtain a compound of formula I. According to some embodiments of the present invention, the hydroxyl protective agent is a silyl ether protective agent. According to some embodiments of the present invention, the hydroxyl protective agent is selected from trimethylchlorosilane, triethylchlorosilane, tert-butyltrichlorosilane, tert-butyldiphenylchlorosilane, triisopropylchlorosilane, dimethylisopropylchlorosilane, tert-butyldimethylchlorosilane, methyldiisopropylchlorosilane, triisopropylchlorosilane, and tert-butyldimethylsilyl trifluoromethanesulfonic acid. According to some embodiments of the present invention, the reaction between lanosterol and the hydroxyprotectant can be carried out in the presence of an organic solvent. According to some other embodiments of the present invention, the organic solvent is N,N-dimethylformamide or dichloromethane. According to some embodiments of the present invention, the reaction between lanosterol and the hydroxyprotectant is carried out in the presence of an acid binder. According to some other embodiments of the present invention, the acid binder is an organic base or an inorganic base. According to even more embodiments of the present invention, the acid binder is pyridine, imidazole, diisopropylamine, triethylamine, triethanolamine, potassium carbonate, or sodium carbonate. According to some embodiments of the present invention, the reaction between the lanosterol and the hydroxyprotectant can be carried out at a temperature of 25 to 120°C. According to some embodiments of the present invention, the reaction between the crude lanosterol and the hydroxyprotectant can be carried out at a temperature of 60 to 85°C. Another aspect of the present invention provides a method for separating and purifying a compound of formula I, comprising the step of separating and purifying a starting material A containing a compound of formula I and a compound of formula I' by column chromatography to obtain a compound of formula I. According to some embodiments of the present invention, the mass percentage of compound I in raw material A is 45% to 85%. According