CN113980081B

CN113980081B - Process for the preparation of ergosterol and derivatives thereof

Info

Publication number: CN113980081B
Application number: CN202111334702.XA
Authority: CN
Inventors: 李凯; 唐杰; 谢来宾
Original assignee: Hunan Kerey Pharmaceutical Co ltd
Current assignee: Hunan Kerey Pharmaceutical Co ltd
Priority date: 2021-11-11
Filing date: 2021-11-11
Publication date: 2023-04-07
Anticipated expiration: 2041-11-11
Also published as: CN113980081A

Abstract

The invention discloses a preparation method of ergosterol and derivatives thereof, which comprises the following steps: preparing a wittig reagent, preparing a skeleton steroid structure, and carrying out wittig reaction on the steroid structure to obtain the ergosterol and the derivatives thereof. The invention provides a new preparation idea of ergosterol and derivatives thereof, the ergosterol or the derivatives thereof are obtained by a synthesis mode, a target structure can be obtained in a targeted manner, and the yield and the purity are high. In addition, the invention takes the phytosterol and the alcohol reagent as raw materials, and the raw materials have wide sources and are environment-friendly and suitable for industrial production.

Description

Process for the preparation of ergosterol and derivatives thereof

Technical Field

The invention relates to the technical field of pharmaceutical chemistry synthesis, and more particularly relates to a preparation method of ergosterol and derivatives thereof.

Background

Ergosterol is a precursor for producing vitamin D2 and is an intermediate for producing hormone drugs, and can be converted into vitamin D2 under ultraviolet irradiation. Ergosterol is a plant sterol found in fungi such as yeast and ergot, is an important component of microbial cell membranes, and plays an important role in ensuring the integrity of cell membranes, the activity of membrane-bound enzymes, the fluidity of membranes, the viability of cells, the transportation of cell substances and the like. Studies have shown that ergosterol may have anti-tumor properties.

The ergosterol is extracted from cultured yeast by culturing yeast thalli and crushing, the yield is very low, the waste water amount is large, and the ergosterol extracted by the method contains other homologues, so that the extraction and separation are relatively difficult.

At present, no relevant route of mature synthesis is reported.

Disclosure of Invention

Based on the technical problems in the prior art, the invention provides a preparation method of ergosterol and derivatives thereof, which can be used for pertinently obtaining target structures of ergosterol and derivatives thereof, and has high yield and high purity.

In order to achieve the purpose, the technical scheme of the invention is as follows:

step 1, preparation of wittig reagent:

step 1-1: performing Mannich reaction on the compound A0 under the conditions of dimethylamine hydrochloride and methanol water to obtain a compound A1;

step 1-2: carrying out catalytic hydrogenation on the compound A1 in a methanol solvent, and reducing by using sodium borohydride to obtain a compound A3; in the reaction process, adding a catalyst and a polymerization inhibitor, wherein the catalyst is Raney nickel, and the polymerization inhibitor is 2, 6-di-tert-butyl-p-cresol; the hydrogenation temperature is 30 to 60 ℃;

step 1-3: performing halogenation reaction on the compound A3 to obtain a compound A4;

step 1-4: reacting the compound A4 with a phosphorus reagent to obtain the wittig reagent;

step 2, reacting the compound VI with the wittig reagent to obtain ergosterol and derivatives thereof;

wherein said compound VI has the structure of formula VI, and said ergosterol and derivatives thereof have the structure of formula VII:

(formula VI); />

(formula VII); the R is ₁ Is->

(ii) a The structure of the compound A0 is as follows: />

. Specifically, in step 1, taking compound A0 as isovaleraldehyde as an example, the preparation route of the wittig reagent is as follows:

(ii) a In the preparation of A3 from compound A1, compound A2 and compound A3 are generally formed and are worked up to give compound A3.

In some embodiments, the phosphorus reagent comprises at least one of triphenylphosphine, trimethyl phosphite, triethyl phosphite, and the like; triphenylphosphine is preferably used.

In some embodiments, in step 1-4, the solvent is toluene, and a catalyst is further added to the reaction system, wherein the mass of the catalyst is 1 to 5% of that of the halogenated isopentane, and the catalyst is potassium iodide. In some embodiments, during the reaction of step 1-2, a catalyst and a polymerization inhibitor are added, wherein the catalyst is raney nickel, and the polymerization inhibitor is 2, 6-di-tert-butyl-p-cresol; the hydrogenation temperature is 30 to 60 ℃. Heating in methanol solvent, adding polymerization inhibitor and Raney nickel for catalytic hydrogenation; the reaction rate can be accelerated, the polymerization degree can be reduced, and the yield of the compound A3 can be improved by heating and taking Raney nickel as a catalyst; the polymerization inhibitor is added, so that the polymerization yield can be further reduced, and the yield is further improved; the hydrogenation temperature is 30 to 60 ℃; the preferred hydrogenation temperature is 55 ℃.

In some embodiments, in step 1-4, the solvent is toluene, and a catalyst is further added, wherein the mass of the catalyst is 1-5% of that of the halogenated isopentane, and the catalyst is potassium iodide. Through the test of the inventor, different solvents and moisture have certain influence on the conversion rate, wherein the yield of the toluene in the water is the highest, and the potassium iodide is added as a catalyst, so that the conversion rate can be improved, the generation of isomers can be inhibited, and the yield of a wittig reagent of the compound can be improved.

In some embodiments, compound V is used as a raw material, and side chain hydroformylation is performed to obtain compound VI; wherein the compound V has the following formula V:

(formula V);

wherein R is ₀ Is a leaving group.

In some embodiments, the compound V is oxidized at DMSO at high temperature under basic conditions to provide compound VI.

In some embodiments, the base used is at least one of sodium bicarbonate, pyridine, triethylamine, 3-methylpyridine, DMAP, sodium hydroxide, sodium carbonate; the reaction temperature is 80 to 120 ℃.

In some embodiments, compound V is prepared from compound I via side chain sulfonylation, dehydrogenation, esterification, reductive dehydrogenation; the structural formula of the compound I is shown as the following formula I:

(formula I).

In some embodiments, the preparation of compound V comprises the steps of:

s1, under an alkaline condition, adding toluoyl chloride into the compound I to react to obtain a compound II;

s2, carrying out dehydrogenation reaction on the compound II to obtain a compound III, wherein the dehydrogenation process is etherification, and then adding a dehydrogenation reagent for dehydrogenation;

s3, reacting the compound III with an esterification reagent under the action of a catalyst to obtain a compound IV;

and S4, reducing the compound IV by a boron hydride compound under an alkaline condition to obtain the compound V.

In some embodiments, the basic conditions are the addition of at least one of DMAP, triethylamine, pyridine, collidine; the reaction solvent is at least one of dichloromethane, chloroform, toluene, ethyl acetate and dichloroethane; the reaction temperature is-10 to 50 ℃.

In some embodiments, in step S2, the dehydrogenation reagent is DDQ and/or chloranil.

In some embodiments, in step S3, the esterification reagent is at least one of an acid anhydride, acetyl chloride, isopropyl acetate, and the catalyst is pTS, HCl, H ₂ SO ₄ ，HClO ₄ At least one of, msOH; and/or the reaction temperature is 0 to 85 ℃.

In some embodiments, in step S4, the alkaline condition is the addition of at least one of NaOH, KOH, naOMe, t-BuOK, pyridine, triethylamine, DMAP; and/or, the borohydride compound comprises Ca (BH) ₄ ) ₂ 、NaBH ₄ 、KBH ₄ At least one of (1).

Compared with the prior art, the invention has the following beneficial effects:

the invention provides a preparation method of ergosterol, which comprises the steps of preparing a wittig reagent, designing a steroid skeleton structure, and finally carrying out synthetic reaction on the wittig reagent and the steroid skeleton structure to obtain the ergosterol or the derivatives thereof. The present invention can obtain ergosterol or its derivative with specific structure in high yield (90 wt%) and high purity (over 99%) higher than that of available technology. In addition, in the method, the aldehyde reagent is used as a raw material, so that the method has the advantages of wide material source, short synthetic route, greenness, environmental protection and suitability for industrial production.

In addition, the invention can greatly improve the yield and purity of the intermediate by optimizing the process conditions in the preparation process, thereby improving the purity and yield of the ergosterol and the derivatives thereof and having high raw material conversion rate.

Detailed description of the preferred embodiments

In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein, but rather should be construed as broadly as the present invention is capable of modification in various respects, all without departing from the spirit and scope of the present invention.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

1. Preparation of Wittig reagent

The specific route is shown as follows by taking isovaleraldehyde (compound A0) as a raw material:

(ii) a The preparation route comprises the following steps:

(1) preparation of Compound A1

0.8kg of compound A0,1.144kg of dimethylamine hydrochloride and 1.1kg of formaldehyde aqueous solution (40%) are added into a reaction bottle, uniformly stirred mechanically, heated to 70 ℃, and stirred for reflux reaction for 4-7h. The product to feed ratio was monitored by GC and work-up was started. A steam distillation device is arranged, the temperature of the steam generation device is about 130-150 ℃, the full bubbles of the steam are ensured, and a hard pipe is connected to be empty; the water vapor was introduced into the reaction solution, and the temperature of the reaction solution was about 110 ℃. And supplementing water at proper time. The distillate was a product and water mixture and distilled for about 8h until the distillate was water only. And separating distillate, wherein the organic phase is the product, about 848g, the purity is 99 percent, the mass yield is 106 percent, and the water content is 1.1 percent.

(2) Preparation of Compound A3

6L of methanol, 2kg of Compound A1 and 10g of 2, 6-di-t-butyl-p-cresol were charged into a reaction flask, and stirred uniformly, and 0.2kg of wet Raney nickel was further added. Blowing hydrogen and heating at 55 ℃ for 24h. After the reaction of the starting materials was monitored by GC, the compounds A2 and A3 were generally formed and the work-up was started. Filtering with diatomite pad, adding 6L dichloromethane into the filtrate, cooling to 0-5 deg.C, slowly adding 0.5kg solid sodium borohydride, controlling the temperature to be less than 20 deg.C, and reacting at about 30 deg.C after adding. GC monitors that the reaction of the raw materials is finished (the reaction liquid is filled in diluted acid and then extracted and sent to a sample). Cooling to 0-5 deg.c, slowly adding dilute hydrochloric acid to quench, discharging gas and releasing heat during quenching, and controlling the adding speed and temperature to less than 30 deg.c. Concentrating, controlling temperature to be less than 40 ℃, concentrating to remove methanol, adding water, adding dichloro for extraction twice, washing an organic phase twice with water, washing an aqueous solution of sodium bicarbonate once, drying the organic phase with anhydrous magnesium sulfate, filtering, and concentrating a filtrate at a temperature of below 40 ℃ to obtain a crude product. The crude product is distilled under reduced pressure, the pressure is about 0.1, the temperature is about 120 ℃, the fraction is collected at 60-80 ℃, and the compound A3 is obtained, wherein the compound A3 is 1.76kg, the mass yield is about 80%, the purity is 99%, 10% of mother liquor is high boiling point substances, and most of the mother liquor is high polymer.

Comparative experiments as shown in Table 1 below were carried out according to the above reaction procedure, and the other conditions were the same as in this example.

TABLE 1 Experimental conditions and results for comparative examples

Comparative example	Condition (take 2kg as an example)	Conversion rate%	Molar yield%
				1	0.1W palladium on carbon, 25 ℃ and 70h	85	50
2	0.1W palladium on carbon, 55 degrees, 55h	90	60
				3	0.1W Raney nickel, 55 degree, 24h	95	70
4	0.1W of Raney nickel, 0.5% of 2, 6-di-tert-butyl-p-cresol, 55 degrees, 24h	95	80

As can be seen from Table 1 and the structure, the compound A1 is easy to polymerize due to its structural characteristics, palladium on carbon is slow to react during hydrogenation, although the conversion rate is high, the degree of polymerization is high, the yield of the distilled compound A3 is low, the reaction speed can be increased by heating temperature and Raney nickel can reduce the degree of polymerization and improve the yield, and the polymerization degree can be further reduced and the yield can be improved by adding the polymerization inhibitor 2, 6-di-tert-butyl-p-cresol.

(3) Preparation of Compound A4

Adding 4L of dichloromethane and 1kg of compound A3 into a reaction bottle, adding 1L of dichloro-diluted 0.5L of phosphorus tribromide into a constant-pressure funnel, cooling to 0-5 ℃, slowly adding phosphorus tribromide, controlling the temperature to be less than 20 ℃, and reacting for 16 hours at about 30 ℃ after the addition is finished. And (3) monitoring by GC (carrying out GC (gas chromatography) to monitor that the reaction solution is washed twice by water and then is subjected to sample sending after being washed by sodium bicarbonate), cooling by 0-5 ℃, slowly adding water for quenching, releasing heat in the quenching process, paying attention to the dropping speed and controlling the temperature to be less than 30 ℃, washing by water for three times, and washing by using sodium bicarbonate solution until the reaction solution is alkalescent. Drying the organic phase, filtering, and concentrating the filtrate to obtain crude product. The crude product is distilled under reduced pressure at about 0.1 and 120 ℃, and the fraction is collected at 60-80 ℃ to obtain compound A4,1.3g, the mass yield is about 130 percent, and the purity is 95 percent.

(4) Preparation of Compound A5 (ylide reagent)

In a 1000mL three-necked flask, 500mL of toluene were added, followed by 100g of Compound A4,1g of potassium iodide and 100g of triphenylphosphine. And (5) carrying out reflux reaction for 24 hours under the protection of nitrogen by dividing water. TLC detection is carried out to ensure that the reaction is basically complete, the solvent is removed by concentration, a proper amount of petroleum ether is added, the mixture is stirred for 1 hour, and then the triphenylphosphine remained in the reaction is removed by filtration. A filter cake was obtained as ylide reagent.

Comparative tests as shown in Table 2 below were carried out according to the above reaction procedure, and the other conditions were the same as in this example.

TABLE 2 Experimental conditions and results of comparative examples

Comparative example	Condition	Conversion rate%	Molar yield%
				1	Solvent-free	50	/
2	Acetonitrile (ACN)	55	/
				3	Water separation of toluene	80	75
4	Toluene water diversion, potassium iodide catalysis	90	85

As shown in Table 2, different solvents and water contents for preparing wittig reagent have influence on the conversion rate, wherein the yield of toluene water is the highest, and the addition of potassium iodide catalyst can improve the conversion rate, inhibit the generation of isomer and improve the yield.

2. Preparation of Compound VI

Ergosterol is used as a raw material, and the route is as follows:

(ii) a The preparation route comprises the following steps:

s1, preparation of Compound I

The phytosterol is fermented by microorganism to obtain a compound I.

S2, preparation of Compound II

100g of Compound 1, 5g of DMAP, 100mL of triethylamine and 500mL of DCM were added to a reaction flask at room temperature, replaced with nitrogen, and stirred until the solution was clear. Heating and refluxing. 75g of p-toluenesulfonyl chloride in DCM (200 mL) is added into the system dropwise, and after the dropwise addition is finished for about 30min, the reflux reaction is continued for 1-2h. TLC monitored the reaction until the starting material disappeared. After the reaction, the system was cooled to 10 ℃,40 mL of 50% aqueous methanol solution was added dropwise to quench the reaction, 300mL of water was then added, the solution was separated, and the organic layer was washed with water. Concentrate under reduced pressure to remove most of the solvent, add the appropriate amount of methanol, and continue to concentrate until DCM is completely removed. Keeping about 100mL of methanol, cooling to 0-10 ℃, crystallizing for 1h under stirring, performing suction filtration, leaching with methanol, and drying at 45-50 ℃ to obtain a compound II. The weight yield is about 140 percent, and the purity is more than 98 percent.

Through detection: ¹ H NMR(400MHz ,CDCl3)δ7.75(d ,J＝8.1Hz , 2H) ,7.32(d ,J＝8.0Hz,2H),5.69(s,1H),3.93(dd ,J＝9 .2 ,2 .9Hz ,1H) ,3 .75(dd ,J＝9.1 ,6.5 Hz ,1H) ,2 .52–2 .16(m ,7H) ,2.06–1 .88

(m ,2H) ,1.86–1 .32(m ,8H) ,1 .18–0 .78(m ,13H) , 0.65(s,3H) 。

s3, preparation of Compound III

Adding 50g of compound II, 250mL of anhydrous methanol, 2.5g of PTS (p-toluenesulfonic acid) and 40mL of trimethyl orthoacetate into a reaction flask under stirring at room temperature, keeping the temperature at 30 ℃ for about 3 hours, after the reaction of the raw materials is finished, adding 200mL of acetone, 35mL of water and 40g of chloranil, slowly heating to about 40 ℃ under stirring for reaction, monitoring by TLC, after about 3-4 hours of reaction, pouring the reaction system into 500mL of water to precipitate solid, filtering, heating the solid to 50 ℃ by 200mL of chloroform for dissolving, filtering while the mixture is hot, heating and dissolving a filter cake by 50mL of chloroform, filtering, and combining organic phases. Adding saturated sodium sulfite aqueous solution (containing 25g of sodium sulfite) into the organic phase, stirring for 1h, standing for layering, concentrating the organic phase under reduced pressure to remove most of the solvent, adding methanol, continuously concentrating (the operation is carried out for 3 times), keeping about 50mL of methanol, cooling to 0 ℃, crystallizing for 1h, carrying out suction filtration, leaching with methanol, and drying at 45-50 ℃ to obtain a compound 3. The weight yield is about 90 percent, and the purity is more than 93 percent.

Through detection: ¹ H NMR(400MHz , CDCl3)δ7 .74(t ,J＝7 .1Hz ,2H) ,7 .32(d ,J＝7 .7Hz ,2H) ,6 .06(d ,J＝6 .6Hz ,2H) ,5 .62(d ,J＝6 .5Hz ,1H) ,3 .94(dd ,J＝9 .0 ,2 .3Hz ,1H) ,3 .84–3 .61(m ,1H) ,2 .64–2 .46(m ,1H) ,2 .45–2 .25 (m ,4H) ,2 .13(t ,J＝10 .1Hz ,1H) ,2 .04–1 .84(m ,2H) ,1 .82–1 .31(m ,6H) ,1 .31–1 .02(m ,9H) , 1 .04–0 .89(m ,3H) ,0 .73–0 .58(m ,3H)。

s4, preparation of Compound IV

At room temperature, adding 50g of compound III, 250mL of acetic anhydride and 100mL of acetyl chloride into a reaction bottle, heating to reflux reaction in a dark place, and monitoring by TLC after about 6-8h, wherein the residual amount of the raw materials is less than 5%. Concentrating the reaction solution at about 75 ℃ under reduced pressure until the reaction solution is dry, cooling to room temperature, dropwise adding 25mL of methanol to quench the residual acetic anhydride, adding 50mL of acetone, concentrating under reduced pressure to remove most of the solvent, adding 100mL of acetone and continuing to concentrate, keeping about 50mL of acetone, cooling to 0 ℃ for crystallization for 1h, filtering, leaching with glacial acetone, and drying the solid at 45-50 ℃ to obtain the compound VI, wherein the weight yield is about 90%, and the purity is more than 95%.

Through detection: 1H NMR (400 MHz, CDCl 3) δ 7.76 (d, J = 7.8 Hz, 2H), 7.32 (d, J = 7.9 Hz, 2H), 5.73 (s, 1H), 5.55 (d, J = 5.7 Hz, 1H), 5.47 (s, 1H), 3.95 (dd, J = 9.2, 2.3 Hz, 1H), 3.85-3.69 (m, 1H), 2.54 (dd, J = 21.0, 8.4 Hz, 1H), 2.42 (s, 3H), 2.19-1.96 (m, 6H), 1.87 (dd, J = 12.4, 5.3 Hz, 2H), 1.77-1.49 (m, 6H), 1.48-1.14 (m, 4H), 1.03-0.91 (m, 6H), 0.56 (s, 3H).

S5, preparation of Compound V

At room temperature, adding 3.5g of anhydrous calcium chloride, 20g of pyridine, 200mL of methanol and 200mL of THF into a reaction bottle, and stirring for dissolving; then cooling to-10-15 ℃, adding sodium borohydride into 4 batches at an interval of 10min for 2g each time, adding 50g of compound IV after all the sodium borohydride are added, keeping the system temperature at-5-10 ℃ for reaction after the sodium borohydride is added, reacting for about 8-10h, and monitoring by TLC (thin layer chromatography) until no raw material is left; and slowly pouring the reaction solution into 500mL of ice water, stirring while adding, stirring for 20min after the solid is separated out, slowly dropwise adding 10mL of glacial acetic acid into the system, filtering, and leaching with water. Dissolving the solid with 150mL of DCM, separating out the water layer, concentrating the organic phase under reduced pressure to remove most of the solvent, adding methanol and continuing to concentrate (the operation is carried out for 3 times), finally retaining about 50mL of methanol, cooling to 0 ℃ for crystallization for 1h, carrying out suction filtration, rinsing with ice methanol, and drying at 45-50 ℃ to obtain the compound V. The weight yield is about 70 percent, and the purity is more than 95 percent.

And (3) detection: 1H NMR (400 MHz, CDCl) ₃ )δ7 .77(d ,J＝8 .1Hz ,2H) ,7 .33(d ,J ＝8 .0Hz ,2H) ,5 .54(d ,J＝3 .7Hz ,1H) ,5 .44–5 .20(m ,1H) ,3 .95(dd ,J＝9 .2 ,2 .7Hz ,1H) ,3 .80 (dd ,J＝9 .1 ,6 .4Hz ,1H) ,3 .67–3 .52(m ,1H) ,2 .58–2 .37(m ,4H) ,2 .26(t ,J＝12 .8Hz ,1H) , 2 .08–1 .78(m ,6H) ,1 .74–1 .14(m ,11H) ,0 .98(dd ,J＝11 .3 ,6 .1Hz ,3H) ,0 .87(d ,J＝31 .0Hz , 3H) ,0 .59(d ,J＝23 .9Hz ,3H) .

13C NMR (101 MHz, CDCl 3) delta 144.59, 140.44, 140.05, 133.01, 129.72, 127.84, 119.39, 116.57, 75.49, 70.26, 53.97, 51.43, 46.03, 42.94, 40.67, 38.77, 38.27, 36.94, 36.44, 31.84, 27.26, 22.93, 21.57, 20.94, 16.92, 16.19, 11.70. (2 carbon signals overlap with other signals)

S6, preparation of Compound VI

Adding 50g of compound V into a reaction bottle at room temperature, adding 150mL of anhydrous DMSO (dimethyl sulfoxide), adding 22mL of triethylamine while stirring, heating to 100 ℃ for reaction for 2-3h, and monitoring the completion of the reaction of raw materials by TLC; slowly pouring into 1L of water for water separation, performing suction filtration, leaching with water, heating and pulping the crude product with 100mL of methanol for 1-2h, cooling to 0 ℃ for crystallization for 1h, performing suction filtration, leaching with ice methanol, and drying at 45-50 ℃ to obtain a compound VI. The weight yield is about 60 percent, and the purity is more than 95 percent.

Comparative experiments as shown in Table 3 below were carried out according to the above reaction procedure, and the other conditions were the same as in this example.

TABLE 3 Experimental conditions and results for comparative examples

Comparative example	Condition	Conversion rate%	Molar yield%
				1	22eq sodium bicarbonate, DMSO (2% moisture)	50	-
2	1.5eq sodium bicarbonate, DMSO (2% moisture)	50	-
				3	1.5eq pyridine, DMSO (2% moisture)	75	70
4	1.5eq Triethylamine, DMSO (2% moisture)	80	75
				5	1.5eq Triethylamine, DMSO (moisture)<0.1%）	93	85

As shown in Table 3, the reaction requires the water content and the base type, when the water content is high, the hydrolysis impurity can reach 15-20%, while the base type is different, the isomer impurity difference of the reaction is obvious, about 2-30%, generally, the inorganic base isomer is large, the organic base isomer is small, and the current optimum conditions are 1.5eq triethylamine and DMSO (the water content is less than 0.1%).

3. Preparation of ergosterol

Taking the prepared compound VI as a raw material, and carrying out wittig reaction with the prepared compound A5 to obtain ergosterol, wherein the route is as follows:

(ii) a In particular toThe method comprises the following steps:

adding 500mL of anhydrous THF into a 1000mL three-necked flask, adding the prepared ylide reagent A5, dropwise adding 150mL of n-butyllithium at 0 ℃ under the protection of nitrogen, controlling the temperature to be below 10 ℃, adding 50g of the compound VI, and slowly raising the temperature to the normal temperature for reaction for 1h. The reaction was monitored by TLC. After the reaction, 25mL of water was added dropwise to quench the reaction. Concentrating to remove THF, adding 250mL of water, extracting the water phase with petroleum ether (200mL x 2), combining the organic phases, concentrating, adding methanol, continuing to concentrate to a small volume, stirring, cooling to 0-5 ℃, crystallizing for 1 hour, and filtering to obtain a crude product. Adding 400mL of absolute ethyl alcohol and 0.5g of activated carbon into the crude product, stirring and decoloring at 30-35 ℃ for 2h, filtering, concentrating at 30-35 ℃ to a small volume, stirring and cooling to 0-5 ℃, crystallizing for 1h, filtering, and drying in vacuum at 30-35 ℃ to obtain a white crystal ergosterol product. The weight yield is about 90 percent, and the purity is more than 99 percent.

And (3) detection: ¹ H NMR(400MHz ,CDCl3)δ5 .56(dd ,J＝10 .6 ,7 .2Hz ,1H) ,5 .37(dd ,J＝14 .1 , 11 .3Hz ,1H) ,5 .26–5 .04(m ,2H) ,3 .71–3 .50(m ,1H) ,2 .45(dt ,J＝30 .1 ,15 .1Hz ,1H) ,2 .26 (dd ,J＝25 .2 ,12 .7Hz ,1H) ,2 .11–1 .18(m ,19H) ,1 .02(t ,J＝7 .4Hz ,3H) ,0 .96–0 .88(m ,6H) , 0 .82(dd ,J＝13 .1 ,6 .7Hz ,6H) ,0 .63(s ,3H) 。

¹³ CNMR(101MHz ,CDCl3)δ141 .35 ,139 .77 ,135 .56 ,131 .98 ,119 .59 ,116 .28 ,70 .47 ,55 .74 ,54 .56 ,46 .26 ,42 .83 ,40 .79 ,40 .41 ,39 .09 ,38 .38 ,37 .04 ,33 .09 ,31 .99 ,28 .28 ,23 .00 ,21 .11 , 19 .95 ,19 .64 ,17 .60 ,16 .28 ,12 .05。

all possible combinations of the technical features of the above embodiments may not be described for the sake of brevity, but should be considered as within the scope of the present disclosure as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent should be subject to the appended claims.

Claims

1. The preparation method of the ergosterol and the derivatives thereof is characterized by comprising the following steps:

step 1, preparation of wittig reagent:

step 1-3: performing halogenation on the compound A3 to obtain a compound A4;

(formula VI);

(formula VII);

the structure of the compound A0 is as follows:

；

in the step 1-1, the specific reaction steps are as follows:

adding 0.8kg of compound A0,1.144kg of dimethylamine hydrochloride and 1.1kg of formaldehyde aqueous solution (40%) into a reaction bottle, mechanically stirring uniformly, heating to 70 ℃, and stirring and refluxing for reaction for 4-7h; GC monitors the ratio of the product to the raw material, and after-treatment is started; a steam distillation device is arranged, the temperature of the steam generation device is 130-150 ℃, the full bubbles of the steam are ensured, and a hard pipe is connected to be empty; introducing water vapor into the reaction solution, and keeping the temperature of the reaction solution at 110 ℃; supplementing water at proper time, wherein the distillate is a mixture of the product and water, and distilling for 8 hours until the distillate is only water.

2. The process for the preparation of ergosterol and its derivatives as claimed in claim 1, wherein the solvent is toluene and the reaction system further comprises a catalyst, wherein the catalyst is potassium iodide in the steps 1-4.

3. A process for preparing ergosterol and its derivatives as claimed in claim 1 or 2, wherein compound V is used as a raw material, and compound VI is obtained by side chain formylation; wherein compound V has the following structure of formula V:

(formula V).

4. A process for the preparation of ergosterol and its derivatives as claimed in claim 3, characterized in that compound V is oxidized at high temperature in DMSO under basic conditions to give compound VI.

5. A process for the preparation of ergosterol and its derivatives as claimed in claim 3, wherein compound V is prepared from compound I by side chain sulfonylation, dehydrogenation, esterification, reductive dehydrogenation; the structural formula of the compound I is shown as the following formula I:

(formula I).

6. A process for the preparation of ergosterol and its derivatives as claimed in claim 5, characterized in that the preparation of compound V comprises the following steps:

s1, adding tosyl chloride into the compound I to react under an alkaline condition to obtain a compound II;

7. A process for the preparation of ergosterol and its derivatives as claimed in claim 6, wherein in step S1, at least one of DMAP, triethylamine, pyridine and collidine is added under basic conditions; the reaction solvent is at least one of dichloromethane, chloroform, toluene, ethyl acetate and dichloroethane.

8. A process for the preparation of ergosterol and its derivatives as claimed in claim 6, wherein in step S2, the dehydrogenation reagent is DDQ and/or chloranil.

9. A process for preparing ergosterol and its derivatives as claimed in claim 6, wherein in step S3, the esterifying reagent is at least one of anhydride, acetyl chloride and isopropyl acetate, and the catalyst is pTS, HCl, H ₂ SO ₄ ，HClO ₄ And MsOH.

10. A process for the preparation of ergosterol and its derivatives as claimed in claim 6, wherein in step S4, the borohydride compound is Ca (BH) ₄ ) ₂ ,NaBH ₄ , KBH ₄ At least one of; the alkaline condition is at least one of NaOH, KOH, naOMe, t-BuOK, pyridine, triethylamine and DMAP.