CN115260276B

CN115260276B - Process for preparing steroid 16, 17-epoxy compound

Info

Publication number: CN115260276B
Application number: CN202210950723.2A
Authority: CN
Inventors: 曾春玲; 陈舟; 屈俊; 吴亚克; 刘喜荣
Original assignee: Hunan Keyixin Biomedical Co ltd
Current assignee: Hunan Keyixin Biomedical Co ltd
Priority date: 2022-08-09
Filing date: 2022-08-09
Publication date: 2024-06-07
Anticipated expiration: 2042-08-09
Also published as: CN115260276A

Abstract

The invention provides a preparation method of a steroid 16, 17-epoxy compound, which comprises the following steps: by reacting compound ACarrying out an alkynylation reaction to prepare a first intermediate product; carrying out dehydration reaction on the first intermediate product to prepare a second intermediate product, and then carrying out hydrolysis reaction on the second intermediate product in an acidic environment to prepare a third intermediate product; or carrying out Lu Pei rearrangement reaction on the first intermediate product to prepare a third intermediate product; a third intermediate product,And (3) withCarrying out ketal reaction to prepare a fourth intermediate product A; or the third intermediate productCarrying out an etherification reaction to prepare a fourth intermediate product B; subjecting the fourth intermediate A or the fourth intermediate B to olefin epoxidation to prepare a steroid 16, 17-epoxide; the structure of the steroid 16, 17-epoxy compound is shown as any one of the following:

Description

Process for preparing steroid 16, 17-epoxy compound

Technical Field

The invention relates to the field of pharmaceutical chemistry, in particular to a preparation method of a steroid 16, 17-epoxy compound.

Background

Melengestrin acetate, chemical name 17 alpha-acetoxy-6-methyl-16-methylene pregna-4, 6-diene-3, 20-dione (17 alpha-acetoxy-6-methyl-16-methylene-4, 6-prenadiene-3, 20-dione), has antitumor and progestin-like activity, and can be widely used as a growth promoting additive for livestock raising, which can improve raising efficiency, thereby accelerating weight gain of livestock. The structural formula of melengestrol acetate is shown as follows:

In the synthesis and preparation process of melengestrol acetate, the construction of the functional group on the steroid D ring is the most critical. The technical route of the British BDH (British drug houses limited) company, the technical route developed by the Puqiang company, the synthetic preparation process of the melengestrol acetate reported by the patent and journal mostly uses epoxy as an intermediate, and the route of the 16, 17-epoxy compound for synthesizing the melengestrol acetate is shown as follows:

Another technical route for melengestrol acetate developed by the common company is not to use epoxy as an intermediate, but the implementation condition is severe, the individual steps are required to be carried out at low temperature, the post-treatment of some steps is complicated, the yield is low, the individual steps are required to oxidize mercury and dimethyl oxalate by using highly toxic reagents, and the dangerous reagents butyl lithium, and the industrial use of benzene sulfinyl chloride and trimethoxy phosphorus is also more inconvenient. Therefore, the synthesis of melengestrol acetate by using an epoxy compound as an intermediate is an extremely important synthesis means, and 16, 17-epoxy compound is a key intermediate for preparing melengestrol acetate.

The traditional preparation method of the intermediate steroid 16, 17-epoxy compound for the synthesis of melengestrol acetate has the following defects: (1) The starting materials for synthesizing the steroid 16, 17-epoxy compound are not easily available and have high price, for example, the starting materials adopted by the traditional method need to be synthesized by yam saponin through 10 steps; (2) The traditional preparation method needs diazomethane addition step and high-temperature (170 ℃) denitrification gas process on the introduction of 16-position methyl, and the two processes have operation dangers and risk of poisoning and explosion.

Disclosure of Invention

Based on the above, the invention provides a preparation method of steroid 16, 17-epoxy compound, which has the advantages of low cost and easy obtaining of raw materials, high yield and safe and convenient operation.

The invention is realized by the following technical scheme.

A method for preparing a steroid 16, 17-epoxy compound, comprising the steps of:

by reacting compound A Performing an alkynylation reaction to prepare a first intermediate product

Subjecting the first intermediate product to dehydration reaction to prepare a second intermediate productThen the second intermediate product is subjected to hydrolysis reaction in an acidic environment to prepare a third intermediate productOr subjecting the first intermediate product to Lu Pei rearrangement reaction to prepare the third intermediate product;

The third intermediate product, And/>Ketal reaction to produce the fourth intermediate A/>Wherein R ₁ is independently selected from alkyl groups with 2-3 carbon atoms, and R ₂ is independently selected from alkyl groups with 1-3 carbon atoms; or (b)

The third intermediate product is subjected toCarrying out an etherification reaction to prepare a fourth intermediate product

Subjecting the fourth intermediate a or the fourth intermediate B to olefin epoxidation to prepare the steroid 16, 17-epoxide; the structure of the steroid 16, 17-epoxy compound is shown in any one of the following formulas (1) and (2):

in one embodiment, the conditions of the alkynylation reaction include:

Mixing the compound A, acetylene and inorganic base with a first organic solvent, and performing an alkynylation reaction at the temperature of-20 ℃ to 20 ℃ for 1h to 3h; or (b)

Combining said compound A withAnd (3) carrying out an etherification reaction, then mixing a product obtained by the etherification reaction with trimethylsilyl ethynyl lithium, carrying out an alkynylation reaction at the temperature of-78 ℃ to-20 ℃, and then mixing the product obtained by the alkynylation reaction with alkali and acid in sequence.

In one embodiment, the first organic solvent is selected from one or more of tetrahydrofuran and ethanol.

In one embodiment, the conditions for the dehydration reaction include:

Mixing the first intermediate product, phosphorus oxychloride and a second organic solvent, and carrying out dehydration reaction at 60-70 ℃ for 1-10 h; or (b)

Mixing the first intermediate product, copper sulfate and a third organic solvent, and carrying out dehydration reaction at the temperature of 110-130 ℃ for 4-6 h; or (b)

Mixing the first intermediate product with a fourth organic solvent, cooling to-35 ℃ to-15 ℃, mixing with N-chlorosuccinimide and a pyridine solution of sulfur dioxide, and carrying out dehydration reaction.

In one embodiment, the second organic solvent is pyridine;

the third organic solvent is selected from one or more of toluene and xylene;

The fourth organic solvent is pyridine.

In one embodiment, the conditions of the hydrolysis reaction include:

mixing the second intermediate product with organic acid, and carrying out hydrolysis reaction for 0.5-3 h at the temperature of 70-80 ℃; the organic acid is selected from one or two of formic acid and acetic acid; or (b)

Mixing the second intermediate product, hydrochloric acid and ethyl acetate, and carrying out hydrolysis reaction for 0.5-1.5 h at the temperature of 75-85 ℃.

In one embodiment, the conditions for the Lu Pei rearrangement reaction include:

Mixing the first intermediate product with acid, and carrying out Lu Pei rearrangement reaction at the temperature of 80-100 ℃ for 0.5-1.5 h.

In one embodiment, the ketal reaction conditions include:

The ketal reaction is carried out in a fifth organic solvent; the fifth organic solvent is selected from one or more of dichloromethane, tetrahydrofuran, dioxane and ethanol.

In one embodiment, the conditions of the etherification reaction include:

The etherification reaction is carried out in a sixth organic solvent; the sixth organic solvent is selected from one or more of dichloromethane, tetrahydrofuran, dioxane and ethanol.

In one embodiment, the conditions for the olefin epoxidation reaction include:

Mixing hydrogen peroxide, inorganic base and a seventh organic solvent with the fourth intermediate product A or the fourth intermediate product B, and carrying out olefin epoxidation reaction at the temperature of 25-40 ℃ for 24-72 h; the seventh organic solvent is selected from one or more of methanol, ethanol and dichloromethane.

Compared with the prior art, the preparation method of the steroid 16, 17-epoxy compound has the following beneficial effects:

The invention takes 4-AD methyl (androstane-4-alkene-16-methyl-3, 17-diketone) as a starting material to prepare the steroid 16, 17-epoxy compound through four-step or five-step reaction, and the starting compound can be prepared from the cheap raw material 4-AD (androstane-4-alkene-3, 17-diketone) through two steps according to the prior art, and can also be obtained by the market convenience, and is cheap and easy to obtain. Moreover, the preparation method of the invention has the advantages of high yield of the obtained product, few byproducts, direct reaction without purification in the second step to the fourth step, simple operation, no need of using high-toxicity and high-risk reagents in the whole preparation process, convenient recovery of solvents and reagents, and suitability for industrial production.

Drawings

FIG. 1 is a synthetic route for a steroid 16, 17-epoxide compound provided by the invention;

FIG. 2 is a nuclear magnetic pattern of a first intermediate provided by the present invention;

FIG. 3 is a nuclear magnetic pattern of a first intermediate provided by the present invention;

FIG. 4 is a nuclear magnetic pattern of a second intermediate provided by the present invention;

FIG. 5 is a nuclear magnetic pattern of a third intermediate provided by the present invention;

FIG. 6 is a nuclear magnetic resonance spectrum of a byproduct of a third intermediate preparation process provided by the present invention;

FIG. 7 is a nuclear magnetic resonance spectrum of a fourth intermediate B provided by the present invention;

FIG. 8 is a nuclear magnetic pattern of the steroid 16, 17-epoxy compound provided by the invention.

Detailed Description

In order that the invention may be readily understood, a more complete description of the invention will be rendered by reference to the appended drawings. The drawings illustrate preferred embodiments of the invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

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 herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.

Referring to FIG. 1, the present invention provides a method for preparing a steroid 16, 17-epoxy compound, comprising the steps of:

Subjecting the first intermediate product to dehydration reaction to prepare a second intermediate productThen the second intermediate product is subjected to hydrolysis reaction under an acidic environment to prepare a third intermediate product/>Or carrying out Lu Pei rearrangement reaction on the first intermediate product to prepare a third intermediate product;

A third intermediate product, And/>Ketal reaction to produce the fourth intermediate A/>Wherein R ₁ is independently selected from alkyl groups with 2-3 carbon atoms, and R ₂ is independently selected from alkyl groups with 1-3 carbon atoms; or (b)

The third intermediate productCarrying out an etherification reaction to prepare a fourth intermediate product

Subjecting the fourth intermediate A or the fourth intermediate B to olefin epoxidation to prepare a steroid 16, 17-epoxide; the structure of the steroid 16, 17-epoxy compound is shown in any one of the following formulas (1) and (2):

it is understood that in the present invention, the starting material compound A is 4-AD methyl (androsta-4-ene-16-methyl-3, 17-dione) and can be prepared from inexpensive 4-AD (androsta-4-ene-3, 17-dione) as a starting material in two steps, and the yield is high, and the compound A can also be obtained commercially.

In one specific example of this embodiment, the method comprises,Selected from trimethyl orthoformate, triethyl orthoformate or tripropyl orthoformate.

In one specific example of this embodiment, the method comprises,Selected from ethylene glycol or propylene glycol.

In a specific example, the conditions of the alkynylation reaction include:

Mixing the compound A, acetylene and inorganic base with a first organic solvent, and carrying out an alkynylation reaction at the temperature of-20 ℃ to 20 ℃ for 1h to 3h.

In a specific example, the first organic solvent is selected from one or more of tetrahydrofuran and ethanol. More specifically, the first organic solvent is a mixed solvent of tetrahydrofuran and ethanol.

In a specific example, the inorganic base is selected from one or more of potassium hydroxide and potassium tert-butoxide.

In a specific example, the weight ratio of compound a to inorganic base is 1:2.

More specifically, the conditions of the alkynylation reaction include: at the temperature of minus 20 ℃ to 20 ℃, acetylene gas is introduced into an organic solvent containing inorganic alkali, then solution of the compound A is dripped into the mixture, after the mixture reacts for 1 to 3 hours, the pH value is adjusted to be acidic by acid, water is added into the mixture for water precipitation after the concentration and the removal of part of the organic solvent, methanol is used for recrystallization after suction filtration, and the compound 5 is obtained after drying, and the yield is 80 to 85 percent. When starting compound a is 16αmethyl-4-AD, the first intermediate obtained is a mixture containing the diastereoisomer of the methyl group at position 16, most of the 16αmethyl group undergoes a conformational inversion, while the ethynyl group is all in the α configuration. The mixture ratio is temperature dependent and the ratio of 16α and 16β is about 1:4 at 0deg.C.

In a specific example, the conditions of the alkynylation reaction include:

Combining compound A with And (3) carrying out an etherification reaction, then mixing a product obtained by the etherification reaction with trimethylsilyl ethynyl lithium, carrying out an alkynylation reaction at the temperature of-78 ℃ to-20 ℃, and then mixing the product obtained by the alkynylation reaction with alkali and acid in sequence.

Preferably, the base is a carbonate. More specifically, the carbonate is potassium carbonate.

In one specific example, the acid is hydrochloric acid.

In a specific example, the etherification reaction is carried out in an organic solvent with an acid.

More specifically, the organic solvent is selected from one or more of ethanol, tetrahydrofuran and dichloromethane. Preferably, the organic solvent is ethanol.

More specifically, the acid is selected from one or more of p-toluenesulfonic acid, sulfuric acid and hydrogen chloride. Preferably, the acid is p-toluene sulfonic acid.

More specifically, the conditions of the alkynylation reaction include: under the catalysis of organic solvent and acid, the raw material compound A reacts with orthoformate to obtain an alkene etherified product protected by 3-site carbonyl, the obtained alkene etherified product reacts with trimethylsilyl ethynyl lithium at the temperature of between 78 ℃ below zero and 20 ℃ below zero, TMS groups on alkynyl groups are removed by potassium carbonate, and finally 3-site protecting groups are removed by hydrochloric acid in methanol to obtain a first intermediate product, wherein the first intermediate product obtained by the method is basically a product with a 16-methyl configuration.

In a specific example, the conditions of the dehydration reaction include:

Mixing the first intermediate product, phosphorus oxychloride and a second organic solvent, and carrying out dehydration reaction at 60-70 ℃ for 1-10 h.

In a specific example, the second organic solvent is pyridine.

In a specific example, the phosphorus oxychloride is fed in a 1-3 times weight ratio to the first intermediate product. Preferably, the phosphorus oxychloride is fed in a 2-fold weight ratio to the first intermediate product.

In a specific example, the second organic solvent is fed in a ratio of 5 to 8 volumes to the first intermediate product. Preferably, the second organic solvent is fed in a ratio of 6 volumes to the first intermediate product.

In a specific example, the heating temperature is from 60 ℃ to reflux, preferably 70 ℃; the reaction time is 1-10h, preferably at 70℃for 6h.

In one specific example, the second intermediate is prepared by quenching in dilute hydrochloric acid after the dehydration reaction has ended, extracting with ethyl acetate, concentrating to dryness, and pulping with ethanol.

In a specific example, the conditions of the dehydration reaction include:

mixing the first intermediate product, copper sulfate and a third organic solvent, and carrying out dehydration reaction at the temperature of 110-130 ℃ for 4-6 h.

In a specific example, the third organic solvent is selected from one or more of toluene and xylene. Preferably, the third organic solvent is xylene.

In a specific example, the heating temperature is 120 ℃ to reflux, preferably reflux, and the reaction time is preferably 5 hours under reflux.

In a specific example, the amount of copper sulfate is 0.1 to 5 times by weight, preferably 3 times by weight, of the first intermediate product.

Although the traditional dehydration reaction uses less copper sulfate, a water separator is needed to remove the generated water, which is not beneficial to industrial mass production, and more importantly, the raw materials are more left. The invention has better dehydration effect by using excessive copper sulfate, and the copper sulfate can be filtered and recovered. After the reaction is finished, the copper sulfate is filtered and removed, and the organic solvent is spin-dried to obtain yellow to brown solid. The reaction byproduct is mainly a Rupe rearrangement product, namely a third intermediate product of the next step, and the product synthesized by the method can be directly put into the next step without purification.

In a specific example, the conditions of the dehydration reaction include:

Reaction temperature control and drop rate are important because 18 methyl rearrangement reactions tend to occur.

In a specific example, the fourth organic solvent is pyridine.

In a specific example, the fourth organic solvent is added in an amount of 4 to 10 volumes, preferably 5 volumes, of the first intermediate product.

In a specific example, N-chlorosuccinimide is added in an amount of 0.8 to 1 times by weight, preferably 0.84 times by weight, of the first intermediate product.

In a specific example, the pyridine solution of sulfur dioxide has a mass concentration of 30%. More specifically, the pyridine solution of sulfur dioxide is added in an amount of 2 times the volume of the first intermediate product.

More specifically, the conditions for the dehydration reaction include: dissolving the first intermediate product in pyridine, cooling, adding NCS, dropwise adding pyridine sulfate solution, TLC detecting reaction, and quenching the reaction with hydrochloric acid.

In a specific example, the conditions of the hydrolysis reaction include:

Mixing the second intermediate product with organic acid, and carrying out hydrolysis reaction for 0.5-3 h at the temperature of 70-80 ℃; the organic acid is selected from one or two of formic acid and acetic acid.

In a specific example, at least one of sulfuric acid and hydrochloric acid is further added to the hydrolysis reaction. Both sulfuric acid and hydrochloric acid can catalyze and accelerate the reaction.

In a specific example, the reaction temperature is from 70 ℃ to reflux, preferably 80 ℃ in the presence of strong acid catalysis, preferably reflux in the absence of strong acid catalysis, and the reaction time is from 0.5 to 3 hours, preferably 1 hour.

In a specific example, the reaction solvent was spin-dried after completion of the reaction, and a brown oil was obtained after water-out, which was purified as a yellow solid. The third intermediate product synthesized by the method can be directly put into the next step without purification after the reaction solvent is dried by a rotary method.

In a specific example, the conditions of the hydrolysis reaction include:

More specifically, the second intermediate was mixed with a hydrochloric acid-ethyl acetate system and subjected to hydrolysis at a temperature of 80 ℃ for 1h.

In a specific example, both hydrochloric acid and ethyl acetate are added in an amount of 10 volumes of the second intermediate product.

In one specific example, the conditions for the Lu Pei rearrangement reaction include:

mixing the first intermediate product with acid, carrying out Lu Pei rearrangement reaction at 80-100 ℃ for 0.5-1.5 h.

It is understood that in the present invention, the acid added in the Lu Pei rearrangement reaction plays a catalytic role.

More specifically, the conditions for the Lu Pei rearrangement reaction include:

Mixing the first intermediate product with formic acid, carrying out Lu Pei rearrangement reaction at 80-100 ℃ for 0.5-1.5 h.

More specifically, lu Pei rearrangement reaction is completed by 1 hour at 100 ℃ with formic acid as a solvent or by 1 hour at 80 ℃ with formic acid as a solvent and sulfuric acid as a catalyst.

In a specific example, after completion of Lu Pei rearrangement, the third intermediate product is prepared by column chromatography.

In one specific example, the ketal reaction conditions include:

In a specific example, the ketal reaction described above also incorporates an acid. More specifically, the acid is one or more of sulfuric acid, p-toluenesulfonic acid, hydrochloric acid and phosphoric acid. Preferably, the ketal reaction is carried out in a methylene chloride-p-toluenesulfonic acid, methylene chloride-phosphoric acid or ethanol-phosphoric acid system. More preferably, the ketal reaction is carried out in a methylene chloride-p-toluenesulfonic acid system.

In a specific example, after the ketal reaction is completed, triethylamine is added to terminate the reaction, the solvent is dried by spinning, methanol is added to pulp, and the fourth intermediate product A is prepared.

In a specific example, the conditions of the etherification reaction include:

In a specific example, the above-described etherification reaction also adds an acid. More specifically, the acid is one or more of sulfuric acid, p-toluenesulfonic acid, hydrochloric acid and phosphoric acid. Preferably, the etherification reaction is carried out in a methylene chloride-p-toluenesulfonic acid, methylene chloride-phosphoric acid or ethanol-phosphoric acid system. More preferably, the etherification reaction is carried out in a methylene chloride-p-toluenesulfonic acid system.

In one specific example of this embodiment, the method comprises,The amount of (2) added is 0.6 to 2 times by volume, preferably 1 time by volume, of the third intermediate product. More specifically, the amount of triethyl orthoformate added is 0.6 to 2 times by volume, preferably 1 time by volume, of the third intermediate product.

In a specific example, after the etherification reaction of the alkene is finished, triethylamine is added to terminate the reaction, the solvent is dried by spinning, methanol is added to pulp, and the fourth intermediate product B is prepared.

In a specific example, the conditions for the olefin epoxidation reaction include:

Mixing hydrogen peroxide, inorganic base and seventh organic solvent with the fourth intermediate product A or the fourth intermediate product B, and carrying out olefin epoxidation reaction at 25-40 ℃ for 24-72 h.

In a specific example, the seventh organic solvent is selected from one or more of methanol, ethanol, and dichloromethane.

In a specific example, the inorganic base is an aqueous sodium hydroxide solution.

In a specific example, the steroid 16, 17-epoxide is prepared by concentrating after the olefin epoxidation reaction is completed, mixing with water, and performing water separation.

The method for producing the steroid 16, 17-epoxy compound of the present invention is described in further detail below with reference to specific examples. The raw materials used in the following examples are all commercially available products unless otherwise specified. The reaction charge ratio in the present invention is often expressed as a weight-to-volume ratio, which refers to the ratio of the weight of the reaction raw material to the weight or volume of the reaction reagent unless otherwise specified.

Example 1

The embodiment provides a preparation method of a first intermediate product, which specifically comprises the following steps:

Weighing 24g of potassium hydroxide powder, adding 120ml of tetrahydrofuran and 13ml of absolute ethyl alcohol, introducing acetylene gas into the ice bath under stirring for 1h, dropwise adding 12g of 40ml of tetrahydrofuran solution of the initiator compound A, after about half an hour, continuing to introduce acetylene gas and react for 2h, detecting that the reaction is basically complete by TLC, dropwise adding 10% hydrochloric acid to a pH value of 1-2, continuing to stir for 10 min, and concentrating under reduced pressure to remove most of the organic solvent. 150ml of water are added and suction filtration is carried out to obtain a yellow solid. The crude product is added into 24ml of methanol to be pulped for 3 hours, light yellow to white solid is obtained by suction filtration, 10.7g of product is obtained after drying, the yield is 82%, the product is a mixture of 16 alpha and 16 beta methyl, and the nuclear magnetic spectrum is shown in figure 2.

Characterization results are as follows ：¹H NMR(400MHz,CDCl₃)δ5.78(s,1H,4-H),2.63+2.56(s,0.2H+0.8H,21-H),1.20(s,3H,19-H),1.17+1.09(d,0.6H+2.4H,22-H),0.94+0.87(s,0.6H+2.4H,18-H);¹³C NMR(CDCl₃)δ199.58,171.18,123.86,88.98,84.06,83.81,79.37,77.17,73.49,53.51,53.43,48.52,48.35,47.52,47.47,44.63,41.26,38.64,36.05,35.68,34.36,34.00,33.95,32.80,32.70,31.88,31.74,31.42,20.69,20.60,17.48,17.45,17.43,17.13,13.47,13.02.

Example 2

4.41g of trimethylethynyl silane is dissolved in 20ml of tetrahydrofuran, the temperature is reduced to-30 ℃, 18ml of butyllithium (2.5 mol/L) is added dropwise under the protection of nitrogen, the reaction is continued for 1h after the completion of the dropwise addition, and then 8.2g of an etherate of compound A (obtained by catalytic reaction of compound A and triethyl orthoformate in ethanol solvent for 5h through p-toluenesulfonic acid) is added dropwise, and the reaction is continued for 2h after the completion of the dropwise addition. After the reaction, adding a mixed solution of 1.73g of potassium carbonate, 12.5g of water and 30ml of methanol, heating to 25 ℃ for continuous reaction for 2 hours, dropwise adding hydrochloric acid to adjust the pH value to 2-3, stirring overnight at room temperature, concentrating most of the solvent under reduced pressure, adding 40ml of water for water precipitation, separating out yellow solid, suction-filtering and washing with 40ml of water, pulping the obtained yellow solid with 19ml of isopropyl ether, suction-filtering to obtain pale yellow to white solid, drying and weighing 7.10g, and obtaining the yield of 87%. The nuclear magnetic pattern of the product is shown in figure 3 and is 16 alpha methyl configuration.

Characterization results are as follows ：¹H NMR(400MHz,CDCl₃)δ5.73(s,1H),2.63(s,1H),2.49–2.21(m,5H),2.03(ddd,1H),1.86–1.59(m,6H),1.59–1.37(m,4H),1.26–1.20(m,1H),1.19(s,3H),1.17(d,3H),1.08–0.95(m,2H),0.94(s,3H).¹³C NMR(CDCl₃)δ199.63,171.30,123.85,84.05,83.82,77.16,53.51,48.34,47.53,41.25,38.65,36.04,35.64,33.94,32.79,32.70,31.88,31.42,20.59,17.42,17.13,13.02.MS m/z：[M+H⁺]＝327.2.

Example 3

The embodiment provides a preparation method of a second intermediate product, which specifically comprises the following steps:

5g of the first intermediate synthesized in example 1 was dissolved in 50ml of xylene, 15g of copper sulfate was added, and the reaction was heated to reflux under nitrogen for 5 hours and checked by TLC until the reaction was substantially complete. After suction filtration, the filter cake is washed by xylene, the filtrates are combined and spin-dried to obtain yellow solid, and the product is directly put into the next step without purification. Passing through a column or recrystallizing to obtain the pure product of the second intermediate product. The nuclear magnetic pattern of the product is shown in figure 4.

Characterization results are as follows ：¹H NMR(400MHz,CDCl₃)δ5.72(s,1H),3.17(s,1H),2.50–2.22(m,5H),2.13–1.96(m,3H),1.85(s,5H),1.79–1.58(m,4H),1.50(ddd,1H)1.42–1.30(m,2H),1.20(s,3H),1.05(m,3H),0.87(s,3H);¹³C NMR(CDCl₃)δ199.43,170.98,148.48,129.26,123.99,82.44,79.00,55.07,54.32,47.94,38.79,36.80,35.54,34.63,34.22,33.94,32.77,31.82,20.77,17.22,16.60,16.04.MS m/z：[M+H⁺]＝309.2.

Example 4

5g of the first intermediate synthesized in example 1 was dissolved in 50ml of xylene, 15g of copper sulfate was added, and the reaction was heated to reflux under nitrogen for 5 hours and checked by TLC until the reaction was substantially complete. After suction filtration, the filter cake is washed by xylene, the filtrates are combined and spin-dried to obtain yellow solid, and the product is directly put into the next step without purification. Passing through a column or recrystallizing to obtain the pure product of the second intermediate product.

Example 5

Under the protection of nitrogen, 10g of the first intermediate product synthesized in the example 1 and 50ml of pyridine are added, a solution system is obtained by stirring, the temperature is reduced to-20 ℃, 8.4g of NCS is added at about-15 ℃ to-20 ℃, and then the temperature is reduced to-35 ℃. Weighing 20ml of 30% SO ₂/pyridine solution, diluting with 40ml of pyridine, dripping the diluted SO ₂/pyridine solution into a reaction system, heating the system severely, controlling the internal temperature to-25 to-40 ℃ for about 1 hour, keeping the temperature for reaction for 20-30min at-25 to-35 ℃ after dripping, and detecting the reaction to be complete by TLC. After the reaction is finished, pouring the system into hydrochloric acid aqueous solution precooled to below 5 ℃, precipitating a large amount of white solid, stirring for 1 hour at 0-5 ℃, carrying out suction filtration, washing with water, pressing to dry, and obtaining 8.2g of white solid with 86.8% yield.

Example 6

The embodiment provides a preparation method of a third intermediate product, which specifically comprises the following steps:

50ml of 98% formic acid is added into the second intermediate product synthesized in the embodiment 3, a drop of sulfuric acid is dripped into the second intermediate product, the second intermediate product is placed into an oil bath at 80 ℃ and heated for 1h, TLC detection reaction is complete, the formic acid is dried in a spinning way to obtain black to brown oil, water is added into the second intermediate product, brown viscous solid is obtained through water separation, 50ml of dichloro is added into the second intermediate product for dissolution, sodium carbonate is added into the second intermediate product for neutralization to remove residual sulfuric acid and formic acid, the second intermediate product is separated into liquid and dried, solvent is concentrated to obtain brown viscous solid, and the second intermediate product is directly put into the next step without purification. The third intermediate product can be obtained by column passing or recrystallization.

Characterization results are as follows ：¹H NMR(400MHz,CDCl₃)δ5.71(s,1H),2.48–2.31(m,3H),2.29(m,1H),2.26(s,3H),2.21(ddd,1H),2.14(m,2H),2.01(s,3H),1.98(m,1H),1.83(m,1H),1.76–1.55(m,3H),1.49(m,1H),1.30(m,3H),1.19(s,3H),1.14-1.01(m,1H),0.98(s,3H).¹³C NMR(CDCl₃)δ199.44,198.89,170.93,151.74,148.23,123.98,54.07,54.03,48.27,39.65,38.70,35.52,34.96,33.93,33.71,32.72,31.80,31.42,20.82,17.98,17.20,16.12.MS m/z：[M+H⁺]＝327.2.

Example 7

3g of the first intermediate obtained in example 1 was dissolved in 30ml of formic acid, a drop of sulfuric acid was added, and the mixture was heated in an oil bath at 80℃for 1h, and the TLC detection was complete, and the formic acid was dried by spin-drying to give a black to brown oil, and 1.44g of a yellow solid, the third intermediate, was obtained by column chromatography, in 48% yield. The nuclear magnetic pattern of the product is shown in FIG. 5, and the nuclear magnetic pattern of the by-product of column chromatography is shown in FIG. 6.

Example 8

The embodiment provides a preparation method of a fourth intermediate product B, which specifically comprises the following steps:

The compound 3 obtained in example 6 was dissolved in 50ml of methylene chloride, 5ml of triethyl orthoformate was added thereto, 0.05g of p-toluenesulfonic acid was further added thereto, and after stirring at room temperature for 3 hours, 1ml of triethylamine was added thereto to terminate the reaction. The solvent was removed by concentrating under reduced pressure, 25ml of methanol was added, and the solid was beaten by stirring, and a brown solid was obtained by suction filtration, and the obtained brown solid was beaten by adding 10ml of isopropyl ether for 1 hour, and a yellow solid was obtained by suction filtration, 2.21g. The overall yields of the three steps of example 3, example 6 and example 8 were 41%. The nuclear magnetic pattern of the product is shown in FIG. 7.

Characterization results are as follows ：¹H NMR(400MHz,CDCl₃)δ5.21(q,1H),5.12(d,1H),3.78(m,2H),2.28(s,3H),2.03(s,3H),1.30(t,3H),1.01(s,6H);¹³C NMR(CDCl₃)δ199.28,154.64,152.08,148.49,141.50,117.43,98.98,62.18,55.09,48.64,48.36,39.74,35.35,35.20,33.70,31.48,31.39,30.08,25.50,20.93,18.91,17.97,16.18,14.67.MS m/z：[M+H⁺]＝355.3.

Example 9

The embodiment provides a preparation method of a steroid 16, 17-epoxy compound, which comprises the following specific steps:

2.92g of the fourth intermediate B obtained in example 8 was weighed, 45ml of ethanol and 15ml of methylene chloride were added, 9.6ml of sodium hydroxide solution (10%) was further added, the mixture was placed in a water bath at 30℃and 15ml of 30% hydrogen peroxide was added, and the reaction was stirred until the reaction was substantially completed. The reaction solution was concentrated under reduced pressure to remove most of the reaction solution, and then 15ml of water was added thereto for water separation, followed by suction filtration to obtain 2.81g of a yellow solid with a yield of 92%. The nuclear magnetic pattern of the product is shown in figure 8.

Characterization results are as follows ：¹H NMR(400MHz,CDCl₃)δ5.10(s,1H),5.03(s,1H),3.70(m,2H),2.14(s,3H,21-H),1.37(s,3H,22-H),1.22(t,3H,24-H),0.98(s,3H,19-H),0.91(s,3H,18-H);¹³C NMR(CDCl₃)δ206.15,154.57,141.22,117.35,98.91,77.33,68.31,62.17,48.47,46.33,44.40,35.36,33.69,33.31,31.73,31.49,29.84,29.82,25.45,20.55,18.92,16.73,15.43,14.65.MS m/z：[M+H⁺]＝371.3.

Example 10

The embodiment provides a preparation method of a fourth intermediate product A, which specifically comprises the following steps:

1.63g of triethyl orthoformate, 0.66g of ethylene glycol was weighed, 50ml of methylene chloride was added, 0.05g of p-toluenesulfonic acid was added, the mixture was stirred at room temperature for 1 hour, 2.51g of Compound 3 was added, and after 0.5 hour of reaction, 1ml of triethylamine was added to terminate the reaction. Extracting the organic phase twice with water, separating, drying, filtering, concentrating under reduced pressure to remove the solvent, and passing through a column to obtain a fourth intermediate product A.

Example 11

2.92g of the fourth intermediate A obtained in example 10 was weighed, 45ml of ethanol and 15ml of methylene chloride were added, 9.6ml of sodium hydroxide solution (10%) was further added, the mixture was placed in a water bath at 40℃and 15ml of 30% hydrogen peroxide was added, and the reaction was stirred for 40 hours until the reaction was substantially completed. The reaction solution is concentrated under reduced pressure to remove most of the reaction solution, 15ml of water is added, water is separated out, and the yellow solid is obtained by suction filtration, 2.75g, and the yield is 90%.

Characterization results are as follows ：¹H NMR(400MHz,CDCl₃)δ5.72(s,1H),2.51–2.25(m,4H),2.21(s,3H),2.06–1.93(m,1H),1.88–1.74(m,2H),1.73–1.58(m,4H),1.58–1.46(m,2H),1.45(s,3H),1.36(t,1H),1.29–1.21(m,1H),1.19(s,3H),1.14-1.08/1.05-0.93(m,2H),1.07(s,3H).¹³C NMR(101MHz,CDCl₃)δ205.96,199.27,170.48,124.05,76.95,68.20,53.83,45.46,44.32,38.63,35.58,33.88,33.43,33.21,32.67,31.63,31.48,29.78,20.34,17.21,16.66,15.40.MS m/z：[M+H⁺]＝343.28.

The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.

The above examples merely represent a few embodiments of the present invention, which facilitate a specific and detailed understanding of the technical solutions of the present invention, but are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. It should be understood that, based on the technical solutions provided by the present invention, those skilled in the art may obtain technical solutions through logical analysis, reasoning or limited experiments, which are all within the scope of protection of the appended claims. The scope of the patent is therefore intended to be covered by the appended claims, and the description and drawings may be interpreted as illustrative of the contents of the claims.

Claims

1. A process for the preparation of a steroid 16, 17-epoxide comprising the steps of:

The conditions of the dehydration reaction include: mixing the first intermediate product, copper sulfate and a third organic solvent, and carrying out dehydration reaction at the temperature of 110-130 ℃ for 4-6 h; or (b)

2. The method for producing a steroid 16, 17-epoxy compound according to claim 1, wherein the condition of the alkynylation reaction comprises:

3. The method for producing a steroid 16, 17-epoxy compound according to claim 2, wherein the first organic solvent is selected from one or both of tetrahydrofuran and ethanol.

4. The method for producing a steroid 16, 17-epoxy compound according to claim 2, characterized in that the inorganic base is selected from potassium hydroxide.

5. A process for the preparation of a steroid 16, 17-epoxide according to claim 1, characterized in that,

The third organic solvent is selected from one or two of toluene and xylene;

The fourth organic solvent is pyridine.

6. The method for producing a steroid 16, 17-epoxy compound according to claim 1, wherein the conditions for the hydrolysis reaction include:

7. The method for producing a steroid 16, 17-epoxy compound according to any one of claims 1 to 5, characterized in that the conditions for the Lu Pei rearrangement reaction include:

8. The method for producing a steroid 16, 17-epoxy compound according to any one of claims 1 to 6, wherein the ketal reaction conditions include:

9. The process for producing a steroid 16, 17-epoxy compound according to any one of claims 1 to 6, wherein the conditions for the etherification reaction include:

10. The process for producing a steroid 16, 17-epoxy compound according to any one of claims 1 to 6, characterized in that the conditions for the olefin epoxidation reaction include: