CN111303210A - Preparation method of tenofovir alafenamide fumarate isomer impurities - Google Patents

Abstract

The invention discloses a preparation method of a fumaric acid propylphenol tenofovir isomer impurity, which comprises a preparation method of 4 isomer impurities, specifically 9- { (R) -2- [ ((R) - { [ (R) -1- (isopropoxycarbonyl) ethyl ] amino } -phenoxyphosphoryl) methoxy ] propyl } adenine, 9- { (R) -2- [ ((R) - { [ (S) -1- (isopropoxycarbonyl) ethyl ] amino } -phenoxyphosphoryl) methoxy ] propyl } adenine, 9- { (R) -2- [ ((S) - { [ (R) -1- (isopropoxycarbonyl) ethyl ] amino } -phenoxyphosphoryl) methoxy ] propyl } adenine and 9- { (S) -2- [ ((R) - { [ (R) -1- (isopropoxycarbonyl) ethyl ] amino } -phenoxyphosphoryl) methoxy ] propyl } adenine A process for the preparation of (isopropoxycarbonyl) ethyl ] amino } -phenoxyphosphoryl) methoxy ] propyl } adenine. The preparation method has the beneficial effects that the preparation method of the propane fumarate tenofovir isomer impurity is simple and feasible, the raw materials are easy to obtain, the condition is mild, the cost is low, the production is facilitated, and the obtained isomer impurity has important significance for preparing high-purity propane fumarate tenofovir.

Description

Preparation method of tenofovir alafenamide fumarate isomer impurities

technical field

The invention relates to the technical field of drug synthesis, in particular to a method for preparing tenofovir fumarate alafenamide isomer impurities.

Background technique

Tenofovir alafenamide fumarate (TAF) is a reverse transcriptase inhibitor developed by Gilead. Clinical data show that the dosage of TAF is one tenth of that of TFD (tenofovir disoproxil fumarate), and the anti-toxic It has the same ability, and has higher safety and tolerability. It is considered to be the best drug for the treatment of hepatitis B so far.

Impurities in drugs refer to substances that have no therapeutic effect or affect the stability and efficacy of drugs, and are even harmful to human health. The impurities contained in drugs are the main factors affecting the purity of drugs. Impurities may cause changes in physical and chemical constants, variation in appearance and properties, and affect the stability of drugs. Therefore, drug impurity inspection is a very important link to control drug purity and improve drug quality.

The isomer impurity of tenofovir fumarate is a process impurity produced in its production process. Its identification and detection are of great significance to the quality of tenofovir fumarate. Therefore, the study of tenofovir fumarate The preparation method of fovir isomer impurities is of great significance.

SUMMARY OF THE INVENTION

The purpose of the present invention is to solve the above-mentioned problems, and designs a kind of impurity preparation method of tenofovir alafenamide fumarate isomer.

To achieve the above objects, the technical solution of the present invention is, a preparation method of tenofovir alafenamide fumarate isomer impurities, including four preparation methods of isomer impurities, which are respectively the preparation method of compound I and the preparation method of compound II , the preparation method of compound III and the preparation method of compound IV, the preparation method of described compound I comprises the following steps:

S1. Tenofovir and triphenyl phosphite are generated by the action of a catalyst under alkaline conditions to generate monophenyl tenofo;

S2, described monophenyl tenofovir generates phosphorus oxychloride monophenyl tenofovir under the action of chlorination reagent;

S3, D-alanine isopropyl ester hydrochloride is freed under alkaline conditions to obtain D-alanine isopropyl ester;

S4, described phosphorus oxychloride monophenyl tenofovir and described D-alanine isopropyl ester are condensed to generate compound I crude product;

S5, adopt L-tartaric acid to split described compound I crude product, obtain compound I;

Wherein the compound I is impurity 9-{(R)-2-[((R)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methane Code for oxy]propyl}adenine, crude compound I is a mixture of compound I and its diastereomers.

As further description of the present invention, the alkali used in the alkaline condition in S1 is selected from one or more of triethylamine, sodium hydroxide, potassium hydroxide, the catalyst described in S1 is selected from DMAP; the chloride described in S2 The reagent is selected from one or more of thionyl chloride, chlorine, and NCS; the base used in the alkaline condition in S3 is selected from one or more of potassium carbonate, sodium carbonate, potassium bicarbonate, and sodium bicarbonate.

As a further description of the present invention, the preparation method of the compound II comprises the following steps:

S1, monophenyl tenofovir generates phosphorus oxychloride monophenyl tenofovir under the action of chlorination reagent;

S2, L-alanine isopropyl hydrochloride is freed under alkaline conditions to obtain L-alanine isopropyl ester;

S3, described phosphorus oxychloride monophenyl tenofovir and described L-alanine isopropyl ester condensation generates compound II crude product;

S4, adopt L-tartaric acid to split described compound II crude product, obtain compound II;

Wherein the compound II is impurity 9-{(R)-2-[((R)-{[(S)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methane Code for oxy]propyl}adenine, the crude compound II is a mixture of compound II and its diastereomers.

As a further description of the present invention, the chlorinating reagent described in S1 is selected from one or more of thionyl chloride, chlorine, NCS; the alkali used in the alkaline condition in S2 is selected from potassium carbonate, sodium carbonate, hydrogen carbonate One or more of potassium and sodium bicarbonate.

As a further description of the present invention, the preparation method of the compound III comprises the following steps:

S1, monophenyl tenofovir generates phosphorus oxychloride monophenyl tenofovir under the action of chlorination reagent;

S2, D-alanine isopropyl hydrochloride is freed under alkaline conditions to obtain D-alanine isopropyl ester;

S3, described phosphorus oxychloride monophenyl tenofovir and described D-alanine isopropyl ester are condensed to generate compound III crude product;

S4, recrystallize the compound III to obtain compound III;

Wherein the compound III is impurity 9-{(R)-2-[((S)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methane Code for oxy]propyl}adenine, the crude compound III is a mixture of compound III and its diastereomers.

As a further description of the present invention, the chlorinating reagent described in S1 is selected from one or more of thionyl chloride, chlorine, and NCS; the solvent used during the recrystallization in S4 is selected from a mixed solvent of toluene and acetonitrile.

As a further description of the present invention, the volume ratio of toluene and acetonitrile in the mixed solvent is 1:3-1:5.

As a further description of the present invention, the preparation method of the compound IV comprises the following steps:

S1. Tenofovir isomers and triphenyl phosphite generate monophenyl tenofovir isomers by the action of catalysts under alkaline conditions;

S2, the monophenyl tenofovir isomers generate phosphorus oxychloride monophenyl tenofovir isomers under the action of a chlorinating reagent;

S3, D-alanine isopropyl ester hydrochloride is freed under alkaline conditions to obtain D-alanine isopropyl ester;

S4, the phosphorus oxychloride monophenyl tenofovir isomer is condensed with the D-alanine isopropyl ester to generate compound IV crude product;

S5, adopt D-tartaric acid to split described compound IV crude product, obtain compound IV;

Wherein the compound IV is impurity 9-{(S)-2-[((R)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methane Code for oxy]propyl}adenine, crude compound IV is a mixture of compound IV and its diastereomers.

As a further description of the present invention, the alkali used in the alkaline condition in S1 is selected from one or more of triethylamine, sodium hydroxide, and potassium hydroxide; the chlorinating reagent described in S2 is selected from thionyl chloride, One or more of chlorine and NCS; the alkali used in the alkaline condition in S3 is selected from one or more of potassium carbonate, sodium carbonate, potassium bicarbonate, and sodium bicarbonate.

The beneficial effect is that the present invention provides a method for preparing four isomer impurities of tenofovir fumarate, wherein the four isomer impurities are respectively 9-{(R)-2-[(((R )-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methoxy]propyl}adenine, 9-{(R)-2-[( (R)-{[(S)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methoxy]propyl}adenine, 9-{(R)-2- [((S)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methoxy]propyl}adenine and 9-{(S)- 2-[((R)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methoxy]propyl}adenine, the 4 impurities The preparation method is simple and feasible, the raw materials are readily available, the conditions are mild, the cost is low, and the production is favorable.

Description of drawings

Fig. 1 is the 9-{(R)-2-[((R)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methyl of the present invention 1H NMR spectrum of oxy]propyl}adenine;

Fig. 2 is the 9-{(R)-2-[((R)-{[(S)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methyl of the present invention 1H NMR spectrum of oxy]propyl}adenine;

Fig. 3 is the 9-{(R)-2-[((S)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methane of the present invention 1H NMR spectrum of oxy]propyl}adenine;

Fig. 4 is the 9-{(S)-2-[((R)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methyl of the present invention 1H NMR spectrum of oxy]propyl}adenine;

Fig. 5 is the 9-{(R)-2-[((R)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methane of the present invention The synthetic route of oxy]propyl}adenine;

Fig. 6 is the 9-{(R)-2-[((R)-{[(S)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methyl of the present invention The synthetic route of oxy]propyl}adenine;

Fig. 7 is the 9-{(R)-2-[((S)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methan of the present invention The synthetic route of oxy]propyl}adenine;

Fig. 8 is the 9-{(S)-2-[((R)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methane of the present invention Synthetic scheme of oxy]propyl}adenine.

Detailed ways

Tenofovir fumarate is considered to be the best drug for the treatment of hepatitis B so far, because the impurities contained in the drug are the main factors affecting the purity of the drug. The constant changes, the appearance properties vary, and the stability of the drug is affected, so the isomer of tenofovir fumarate is of great significance for the preparation of high-purity tenofovir fumarate. The present invention provides fumaric acid Preparation method of tenofovir alafenamide isomer impurities.

For the convenience of the following description, the following codes will be used to represent the corresponding materials:

Compound I: 9-{(R)-2-[((R)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methoxy]propane base} adenine;

Compound II: 9-{(R)-2-[((R)-{[(S)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methoxy]propane base} adenine;

Compound III: 9-{(R)-2-[((S)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methoxy]propane base} adenine;

Compound IV: 9-{(S)-2-[((R)-{[(R)-1-(isopropoxycarbonyl)ethyl]amino}-phenoxyphosphoryl)methoxy]propane base} adenine.

The invention provides methods for preparing four isomer impurities of tenofovir fumarate, which are respectively a method for preparing compound I, a method for preparing compound II, a method for preparing compound III and a method for preparing compound IV, And the synthetic route diagram of each preparation method is shown in accompanying drawing 5-8, each preparation method will be further described below.

1. The preparation method of compound I comprises the following steps:

S1, tenofovir, triphenyl phosphite and the corresponding solvent are catalyzed by a catalyst to react under alkaline conditions, the reaction temperature is 90-100 ° C, and the reaction time is 8 hours to generate monophenyl tenofovir; Wherein the volume ratio of tenofovir and the solvent is 1:0.2-1:1, the molar ratio of tenofovir and triphenyl phosphite is 1:1-1:2, and the ratio of tenofovir and alkali is 1:1-1:2. The molar ratio is 1:1-1:2, and the molar ratio of tenofovir to the catalyst is 1:1-1:3;

S2, monophenyl tenofovir and the corresponding solvent are reacted under the action of the chlorinating reagent, the reaction temperature is 70-80 ℃, and the reaction time is 2 hours to generate phosphorus oxychloride monophenyl tenofovir; The volume ratio of phenyl tenofovir to the solvent is 1:5-1:15, and the molar ratio of monophenyl tenofovir to the chlorinating reagent is 1:1-1:5;

S3, D-alanine isopropyl hydrochloride and the corresponding solvent are freed under alkaline conditions, and the reaction temperature is 15-25 ℃, and the reaction time is 19 hours to generate D-alanine isopropyl ester; Wherein the volume ratio of D-alanine isopropyl hydrochloride and solvent is 1:2-1:5, and the mol ratio of D-alanine isopropyl hydrochloride and alkali is 1:1-1:3;

S4, phosphorus oxychloride monophenyl tenofovir, D-alanine isopropyl ester and corresponding solvent carry out condensation reaction, temperature of reaction is ^- 20--10 ℃, reaction times is 2 hours, obtains compound I crude product, wherein The crude product of compound I is a mixture of compound I and its diastereomer compound III; the volume ratio of phosphorus oxychloride monophenyl tenofovir to solvent is 1:5-1:10, phosphorus oxychloride monophenyl tenofovir The molar ratio of Wei and D-alanine isopropyl ester is 1:3-1:5;

S5, adopt L-tartaric acid and corresponding solvent to split compound I crude product, and temperature of reaction is 70-80 ℃, the reaction times is 0.5 hour, obtains compound I; Wherein compound I crude product and solvent volume ratio are 1:5- 1:10, the molar ratio of compound I crude product and L-tartaric acid is 1:0.5-1:2.

The solvent used in above-mentioned steps is all selected from one or more in methylene dichloride, acetonitrile, acetone, N,N-di-methylformamide, tetrahydrofuran, wherein the solvent of S1, S2 and S5 is most preferably acetonitrile, S3 And the solvent of S4 is most preferably methylene dichloride;

The alkali used in the above-mentioned S1 is selected from one or more of triethylamine, sodium hydroxide, potassium hydroxide, and triethylamine is most preferably used; the catalyst used in the above-mentioned S1 is most preferably DMAP; the alkali used in the S3 is selected from One or more in potassium carbonate, sodium carbonate, potassium bicarbonate, sodium bicarbonate, preferably potassium bicarbonate; The chlorinating reagent used in above-mentioned S2 is selected from one or more in thionyl chloride, chlorine, NCS species, most preferably thionyl chloride.

2. The preparation method of compound II comprises the following steps:

S1, monophenyl tenofovir and the corresponding solvent are reacted under the action of chlorinating reagent, the reaction temperature is 70-80 ° C, and the reaction time is 2 hours to generate phosphorus oxychloride monophenyl tenofovir; wherein monophenyl tenofovir; The volume ratio of phenyl tenofovir to the solvent is 1:5-1:15, and the molar ratio of monophenyl tenofovir to the chlorinating reagent is 1:1-1:5;

S2, L-alanine isopropyl hydrochloride and corresponding solvent are freed under alkaline conditions, and the reaction temperature is 15-25 ℃, and the reaction time is 19 hours to obtain L-alanine isopropyl ester; Wherein the volume ratio of L-alanine isopropyl ester hydrochloride and solvent is 1:2-1:5, and the mol ratio of L-alanine isopropyl ester hydrochloride and alkali is 1:1-1:3;

S3, phosphorus oxychloride monophenyl tenofovir, L-alanine isopropyl ester and corresponding solvent carry out condensation reaction, the reaction temperature is ^-20--10 ℃, and the reaction time is 2 hours, obtains compound II crude product, wherein The compound II is a mixture of compound II and its diastereomer; the volume ratio of phosphorus oxychloride monophenyl tenofovir and solvent is 1:5-1:10, phosphorus oxychloride monophenyl tenofovir and L -Alanine isopropyl ester molar ratio is 1:3-1:5;

S4, adopt L-tartaric acid and corresponding solvent to split compound II crude product, and reaction temperature is 70-80 ℃, and the reaction time is 2 hours, obtains compound II; Wherein compound II crude product and solvent volume ratio are 1:5- 1:10, the molar ratio of compound II crude product and L-tartaric acid is 1:0.5-1:2.

The solvent in the above steps is selected from one or more of toluene, dichloromethane, acetonitrile, acetone, N,N-di-methylformamide, tetrahydrofuran, wherein S1 and S4 are optimally selected acetonitrile, and S2 and S3 are the most optimal. Dichloromethane is preferred;

The chlorinating reagent in above-mentioned S1 is selected from one or more in thionyl chloride, chlorine, NCS, most preferably thionyl chloride; The alkali in above-mentioned S2 is selected from potassium carbonate, sodium carbonate, potassium bicarbonate, carbonic acid One or more of sodium hydrogen carbonate, potassium hydrogen carbonate is the best choice.

3. The preparation method of compound III comprises the following steps:

S1, monophenyl tenofovir and the corresponding solvent are reacted under the action of chlorinating reagent, the reaction temperature is 80-90 ℃, and the reaction time is 25 hours to generate phosphorus oxychloride monophenyl tenofovir; wherein monophenyl tenofovir; The volume ratio of phenyl tenofovir to the solvent is 1:5-1:15, and the molar ratio of monophenyl tenofovir to the chlorinating reagent is 1:1-1:5;

S2, D-alanine isopropyl hydrochloride and the corresponding solvent are freed under alkaline conditions, and the reaction temperature is 15-25 ° C, and the reaction time is 19 hours to generate D-alanine isopropyl ester; Wherein the volume ratio of D-alanine isopropyl hydrochloride and solvent is 1:2-1:5, and the mol ratio of D-alanine isopropyl hydrochloride and alkali is 1:1-1:3;

S3, phosphorus oxychloride monophenyl tenofovir, D-alanine isopropyl ester and corresponding solvent carry out condensation reaction, the reaction temperature is ^-20--10 ℃, and the reaction time is 2 hours, generates compound III crude product, wherein The crude product of compound III is the mixture of compound III and its diastereomer compound I; wherein the volume ratio of phosphorus oxychloride monophenyl tenofovir and solvent is 1:5-1:10, phosphorus oxychloride monophenyl tenofovir The molar ratio of Wei and D-alanine isopropyl ester is 1:3-1:5;

S4. Recrystallize the crude compound III to obtain compound III.

In the above-mentioned S1, the solvent is selected from one or more of toluene, acetonitrile, acetone, N,N-di-methylformamide, tetrahydrofuran, and toluene is optimally selected; in S2 and S3, the solvent is selected from methylene chloride, acetonitrile , one or more in acetone, toluene, tetrahydrofuran, all optimally select methylene chloride; When carrying out recrystallization in S4, solvent selects toluene and acetonitrile mixed solvent for use, and toluene and acetonitrile volume ratio are 1:3-1: 5;

In the above-mentioned S1, the chlorinating reagent is selected from one or more of thionyl chloride, chlorine, and NCS, and thionyl chloride is optimally selected.

4. The preparation method of compound IV comprises the following steps:

S1, tenofovir isomer, triphenyl phosphite and corresponding solvent are catalyzed by catalyst to react under alkaline conditions, the reaction temperature is 90-100 ℃, and the reaction time is 8 hours to generate monophenyl tenofovir isomer The volume ratio of tenofovir isomer and solvent is 1:0.2-1:1, the molar ratio of tenofovir isomer and triphenyl phosphite is 1:1-1:2, and the tenofovir isomer is 1:1-1:2. The molar ratio to base is 1:1-1:2, and the molar ratio of tenofovir isomer to catalyst is 1:1-1:3;

S2, the monophenyl tenofovir isomer and the corresponding solvent are reacted under the action of the chlorinating reagent, the reaction temperature is 70-80 ℃, and the reaction time is 2 hours, and the phosphorus oxychloride monophenyl tenofovir isomer is generated; Wherein the volume ratio of monophenyl tenofovir isomer and solvent is 1:5-1:15, and the molar ratio of monophenyl tenofovir isomer and chlorinating reagent is 1:1-1:5;

S3, D-alanine isopropyl hydrochloride and corresponding solvent are freed under alkaline conditions, and the reaction temperature is 15-25 ℃, and the reaction time is 19 hours to obtain D-alanine isopropyl ester; Wherein the volume ratio of D-alanine isopropyl hydrochloride and solvent is 1:2-1:5, and the mol ratio of D-alanine isopropyl hydrochloride and alkali is 1:1-1:3;

S4, phosphorus oxychloride monophenyl tenofovir isomer, D-alanine isopropyl ester and corresponding solvent carry out condensation reaction under alkaline conditions, the reaction temperature is -20 ^- -10 ℃, and the reaction time is 2 hours, Generate compound IV crude product, wherein compound IV crude product is a mixture of compound IV and its diastereomer; phosphorus oxychloride monophenyl tenofovir isomer and solvent volume ratio is 1:5-1:10, phosphorus oxychloride monophenyl The molar ratio of tenofovir isomer and D-alanine isopropyl ester is 1:3-1:5;

S5, adopt D-tartaric acid and corresponding solvent to split compound IV crude product, and temperature of reaction is 70-80 ℃, the reaction times is 0.5 hour, obtains compound IV; Wherein compound IV crude product and solvent volume ratio are 1:5- 1:10, the molar ratio of compound IV crude product and D-tartaric acid is 1:0.5-1:2.

In above-mentioned steps, solvent is selected from one or more in toluene, acetonitrile, methylene dichloride, acetone, N,N-di-methylformamide, tetrahydrofuran, wherein S1, S2 and S5 are most preferably acetonitrile, S3 and S4 Dichloromethane is most preferred;

Alkali in above-mentioned S1 is selected from one or more in triethylamine, sodium hydroxide, potassium hydroxide, most preferably uses triethylamine; In S2, chlorinating reagent is selected from thionyl chloride, chlorine, NCS. One or more, thionyl chloride is most preferred; In S3, the base is selected from one or more of potassium carbonate, sodium carbonate, potassium bicarbonate, and sodium bicarbonate, and potassium bicarbonate is most preferred.

Example 1.

S1. Add 100g tenofovir, 162.1g triphenyl phosphite, 42.6g DMAP, 50ml acetonitrile and 71.0g triethylamine to the 2L reaction flask, start heating, heat up to 90-100°C, and keep the temperature for 8h; reaction After the completion, the temperature was lowered to 40-50°C, the acetonitrile was evaporated under reduced pressure, 750ml of acetone and 250ml of acetonitrile were added, the temperature was controlled to 30-40°C, and concentrated hydrochloric acid was slowly added dropwise until the pH was 2.5-3.5. 30°C, heat preservation for 4 hours, filter, rinse the filter cake with 200ml of acetonitrile; put the filter cake into a 2L reaction flask, add 1000ml of acetonitrile, heat up to 50-60°C, stir for 2h, cool down to 20-30°C, keep warm for crystallization Crystallize for 4h, filter, rinse the filter cake with 200ml acetonitrile; put the filter cake into a 2L reaction flask, add 150ml methanol and 650ml acetonitrile, heat up to 50-60°C, stir for 2h, cool down to 20-30°C, and keep crystallization for 4h , filtered, and the filter cake was rinsed with 200 ml of acetonitrile; the filter cake was dried under reduced pressure at 40-50 °C for 12 h, and weighed to obtain 120.5 g of tenofovir monophenyl;

S2. Add 20g monophenyl tenofovir and 200ml acetonitrile into the 500ml reaction flask, turn on stirring, slowly add 19.6g thionyl chloride dropwise, heat up to 70-80°C after the dropwise addition, keep the reaction for 2h, evaporate under reduced pressure Dry, add 200ml of dichloromethane and stir to obtain the dichloromethane slurry of phosphorus oxychloride monophenyl tenofovir, nitrogen protection for use;

S3. Add 37.2g of D-alanine isopropyl hydrochloride, 150ml of dichloromethane and 55.0g of potassium bicarbonate to 500ml of reaction varieties, start stirring, generate a large amount of gas, control the temperature to 15-25°C and react for 19h, filter, Collect the filtrate, put it into a 500ml reaction flask, heat up to 50-60°C, separate water at normal pressure until the moisture content is lower than 0.1%, obtain a dichloromethane solution of D-alanine isopropyl ester and cool it to 10-20°C, Nitrogen protection backup;

S4. Put the dichloromethane slurry of phosphorus oxychloride monophenyl tenofovir into a 500ml reaction flask, turn on stirring, cool down to -20 ^- -10°C, and add the dichloromethane solution of D-alanine isopropyl ester Add it dropwise to the reaction flask, control the temperature to -20 ^- -10°C, keep the temperature for 2 hours after the dropwise addition, then heat up to 10-20°C, wash the solution in the reaction flask with 200ml of 10% sodium dihydrogen phosphate aqueous solution, and finish the washing. Then carry out liquid separation, keep the organic phase, repeat the above operation 3 times, and evaporate the organic phase to dryness under reduced pressure at 40-50 °C to obtain the crude product of compound I;

S5. get above-mentioned compound I crude product 14g and drop into 250ml reaction flask, add acetonitrile 126ml and L-tartaric acid 4.38g, be warming up to 70-80 ℃, stir 30min, dissolve clear, be slowly cooled to 20-30 ℃, filter, 20ml acetonitrile is poured Wash the filter cake; add the filter cake into a 250ml reaction flask, add 112ml of purified water, heat up to 60°C, filter, heat the filtrate to 55-65°C, stir for 30min, slowly cool down to 20-30°C, keep the temperature for 2h, Filter, wash the filter cake with 20ml of purified water, then put the filter cake into a 100ml reaction flask, add 30ml of purified water, heat up to 50-60°C, stir for 30min, slowly cool down to 20-30°C, keep the temperature for 2 hours, and filter , the filter cake was washed with 20 ml of purified water, the filter cake was dried at 50° C. under normal pressure, and weighed to obtain 2.4 g of the tartrate of compound I.

The prepared compound I was detected by nuclear magnetic resonance, and the hydrogen nuclear magnetic resonance spectrum was as shown in accompanying drawing 1;

The obtained compound I was measured by high performance liquid chromatography, and the detection purity was 94.5%, and the retention time was 39.2min;

The above-mentioned high performance liquid chromatography detection method is: chromatographic column: Waters Xbridge ShieldRP (4.6mm × 150mm, 3.5 μm), filler: octadecylsilane bonded silica gel; Mobile phase A: 20mmol/L dipotassium hydrogen phosphate ( Adjust pH to 6.0 with phosphoric acid) - tetrahydrofuran/acetonitrile (7/3) (99:1); mobile phase B: 20 mmol/L dipotassium hydrogen phosphate (adjust pH to 6.0 with phosphoric acid) - tetrahydrofuran/acetonitrile (7/3) (50:50); flow rate: 1.0ml/min; detection wavelength: 260nm; column temperature: 20°C; injector temperature: 5°C; gradient elution as follows:

time (min) Mobile phase A (%) Mobile phase B (%) 0 100 0 10 85 15 15 85 15 40 70 30 50 55 45 55 30 70 60 30 70 61 100 0 70 100 0

Example 2.

S1. Add 20g monophenyl tenofovir and 200ml acetonitrile to the 500ml reaction flask, turn on stirring, slowly add 19.6g thionyl chloride dropwise, heat up to 70-80°C after the dropwise addition, keep the reaction for 2h, reduce the pressure Evaporate to dryness, add 200 ml of dichloromethane and stir to obtain a dichloromethane slurry of phosphorus oxychloride monophenyl tenofovir, nitrogen protection for use;

S2. Add 37.2g of L-alanine isopropyl hydrochloride, 150ml of dichloromethane and 55.0g of potassium bicarbonate to 500ml of reaction varieties, start stirring, generate a large amount of gas, control the temperature at 15-25°C and react for 19h, filter and collect The filtrate was put into a 500ml reaction flask, heated to 50-60°C, and the water was separated at normal pressure until the moisture content was lower than 0.1% to obtain a dichloromethane solution of L-alanine isopropyl ester and cooled to 10-20°C, nitrogen protection spare;

S3. Put the dichloromethane slurry of phosphorus oxychloride monophenyl tenofovir into a 500ml reaction flask, turn on stirring, cool down to -20 ^- -10°C, and add the dichloromethane solution of L-alanine isopropyl ester Add it dropwise to the reaction flask, control the temperature to -20 ^- -10°C, keep the temperature for 2 hours after the dropwise addition, then heat up to 10-20°C, wash the solution in the reaction flask with 200ml of 10% sodium dihydrogen phosphate aqueous solution, and finish the washing. Then carry out liquid separation, keep the organic phase, repeat the above operation 3 times, and evaporate the organic phase to dryness under reduced pressure at 40-50 °C to obtain 23.4 g of compound II crude product;

S4. Take 22g of the above-mentioned compound II crude product and put it into a 250ml reaction flask, then add 198ml of acetonitrile and 6.93g of L-tartaric acid, heat up to 70-80°C, keep the temperature for 2h, slowly cool down to 20-30°C, and keep the temperature for 1.5h. , filtered, and the filter cake was dried under reduced pressure to obtain 26.2g of massive solid; the above-mentioned solid was transferred to a 500ml reaction flask, 196ml of purified water and 19.6ml of acetonitrile were added, the temperature was raised to 70-80°C, stirred for 10min after dissolving, and slowly cooled to to 0-10°C, keep for 1 hour, filter, wash the filter cake with 20ml of purified water; beat the filter cake with 40ml of purified water at room temperature for 1h, cool to 0-10°C, keep for 30min for crystallization, filter, and purify the filter cake with 20ml Wash with water, repeat the above beating operation 3 times to obtain a gray-green solid, and then dry the gray-green solid under reduced pressure at 40-50° C. for 12 h to obtain 2.0 g of the tartrate of compound II.

The prepared compound II was detected by nuclear magnetic resonance, and the hydrogen nuclear magnetic resonance spectrum was shown in accompanying drawing 2;

The obtained compound II was measured by high-performance liquid chromatography. The high-performance chromatography detection method was the same as that in Example 1. The detected purity was 96.12%, and the retention time was 43.9 min.

Example 3.

S1. Add 20g monophenyl tenofovir and 200ml toluene into a 500ml reaction flask, turn on stirring, slowly add 19.6g thionyl chloride dropwise, heat up to 80-90°C after the dropwise addition, keep the reaction for 25h, evaporate under reduced pressure Dry, add 200ml of dichloromethane and stir to obtain a dichloromethane slurry of phosphorus oxychloride monophenyl tenofovir, nitrogen protection for use;

S2. Add 37.2g D-alanine isopropyl hydrochloride, 150ml dichloromethane and 55.0g potassium bicarbonate to the 500ml reaction flask, turn on stirring, generate a large amount of gas, control the temperature at 15-25°C and react for 19h, filter , collect the filtrate, put it into a 500ml reaction flask, heat it up to 50-60°C, divide the water at normal pressure until the moisture is lower than 0.1%, and obtain the dichloromethane solution of D-alanine isopropyl ester and cool it to 10-20°C , nitrogen protection backup;

S3. put the dichloromethane solution of D-alanine isopropyl ester into a 500ml reaction flask, turn on stirring, cool down ^to -20--10°C, put the dichloromethane slurry of phosphorus oxychloride monophenyl tenofovir Add it dropwise to the reaction flask, control the temperature to -20 ^- -10°C, keep the temperature for 2 hours after the dropwise addition, then heat up to 10-20°C, wash the solution in the reaction flask with 200ml of 10% sodium dihydrogen phosphate aqueous solution, and finish the washing. Then carry out liquid separation, keep the organic phase, repeat the above operation 3 times, and evaporate the organic phase to dryness under reduced pressure at 40-50 °C to obtain 20.6 g of compound III crude product;

S4. Take 20g of the above-mentioned compound III crude product, put it into a 250ml reaction flask, add 80ml of toluene and 20ml of acetonitrile, heat up to 65-70°C, dissolve and then keep stirring for 1h, slowly cool down to 0-10°C, and keep crystallization for 6h , filtered, and the above operation was repeated twice to obtain a white solid, which was then dried under reduced pressure at 40-50° C. for 12 h to obtain 10.2 g of compound III.

The prepared compound III was detected by nuclear magnetic resonance, and the hydrogen nuclear magnetic resonance spectrum was shown in accompanying drawing 3;

The prepared compound III was measured by high-performance liquid chromatography. The high-performance chromatography detection method was the same as that in Example 1. The detected purity was 98.39% and the retention time was 31.1 min.

Example 4.

S1. Add 100g tenofovir isomer, 162.1g triphenyl phosphite, 42.6g DMAP, 50ml acetonitrile and 71.0g triethylamine to the 2L reaction flask, start heating, heat up to 90-100°C, and keep the temperature for 8h; to 40-50°C, evaporate the acetonitrile under reduced pressure, add 750ml of acetone and 250ml of acetonitrile, control the temperature to 30-40°C, slowly add concentrated hydrochloric acid dropwise to pH 2.5 to 3.5, keep stirring for 1h, cool down to 20-30°C, and heat for analysis. Crystallize for 4h, filter, rinse the filter cake with 200ml of acetonitrile; put the filter cake into a 2L reaction flask, add 1000ml of acetonitrile, heat up to 50-60°C, stir for 2h, cool to 20-30°C, keep the temperature for crystallization for 4h, filter, 200ml Rinse the filter cake with acetonitrile; put the filter cake into a 2L reaction flask, add 150ml methanol and 650ml acetonitrile, heat up to 50-60°C, stir for 2h, cool down to 20-30°C, keep for 4h crystallization, filter, rinse with 200ml acetonitrile filter cake; the filter cake was dried under reduced pressure at 40-50°C for 12 hours, and weighed to obtain 122.0 g of monophenyl tenofovir isomers;

S2. Add 20g monophenyl tenofovir isomer and 200ml acetonitrile into the 500ml reaction flask, turn on stirring, slowly add 19.6g thionyl chloride dropwise, heat up to 70-80°C after the dropwise addition, keep the reaction for 2h, reduce the pressure Evaporate to dryness, add 200 ml of dichloromethane and stir to obtain a dichloromethane slurry of phosphorus oxychloride monophenyl tenofovir isomer, nitrogen protection for use;

S3. Add 37.2g of D-alanine isopropyl hydrochloride, 150ml of dichloromethane, 55.0g of potassium bicarbonate to 500ml of reaction varieties, turn on stirring, generate a large amount of gas, control the temperature at 15-25°C and react for 19h, filter, Collect the filtrate, put it into a 500ml reaction flask, heat up to 50-60°C, separate water at normal pressure until the moisture is lower than 0.1%, and obtain a dichloromethane solution of D-alanine isopropyl ester and cool it to 10-20°C, nitrogen protection reserve;

S4. Put the dichloromethane slurry of phosphorus oxychloride monophenyl tenofovir isomer into a 500ml reaction flask, turn on stirring, cool down to -20 ^- -10°C, and put the dichloromethane slurry of D-alanine isopropyl ester into a 500ml reaction flask. The solution was added dropwise to the reaction flask, the temperature was controlled at -20 ^- -10 °C, and the temperature was maintained for 2 h after the dropwise addition; the temperature was raised to 10-20 °C, and the solution in the reaction flask was washed with 200 ml of 10% sodium dihydrogen phosphate aqueous solution. Then carry out liquid separation, retain the organic phase, repeat the above operation 3 times, and evaporate the organic phase to dryness under reduced pressure at 40-50 °C to obtain 18.5 g of compound IV crude product;

S5. get above-mentioned crude product 15g, add in 250ml reaction flask, add 135ml acetonitrile, 3.58g D-tartaric acid, be warming up to 70-80 ℃, keep stirring for 30min, slowly cool down to 20-30 ℃, filter, filter cake with 20ml acetonitrile leaching Wash; add the filter cake to a 250ml reaction flask, add 150ml of purified water, heat up to 55-65°C, filter while hot, cool the filtrate to 20-30°C, keep for 4 hours of crystallization, filter, wash with 5ml of purified water, and filter the filtrate. The cake was added to a 500ml reaction flask, 150ml of purified water, 150ml of dichloromethane were added, the pH was adjusted to 8 to 9 with 5% ammonia water, the liquid was separated, the organic phase was collected, the aqueous phase was extracted with 50ml of dichloromethane, and the organic phases were combined. , dried over anhydrous sodium sulfate, and the organic phase was concentrated to dryness under reduced pressure to obtain 2.1 g of compound IV.

The prepared compound IV is detected by nuclear magnetic resonance, and the hydrogen nuclear magnetic resonance spectrum is shown in accompanying drawing 3;

The obtained compound IV was measured by high performance liquid chromatography, and the detection purity was 94.21%, and the retention time was 28.87min;

The above-mentioned high-performance liquid chromatography detection method is: chromatographic column: CHIRALPAK ADH (250×4.6 mm, 5 μm), filler: amylose-tris(3,5-dimethylphenylcarbamate) coated porous Silica gel microspheres; mobile phase: ethanolamine-n-hexane-methanol-n-propanol (0.1:75:9:16); flow rate: 1.0ml/min; detection wavelength: 260nm; column temperature: 35℃; running time: 45 minutes .

The above technical solutions only represent the preferred technical solutions of the technical solutions of the present invention, and some changes that those skilled in the art may make to some parts of them all reflect the principles of the present invention and fall within the protection scope of the present invention.

Claims (9)

1. The preparation method of the isomer impurities of the propane fumarate tenofovir disoproxil comprises the preparation methods of 4 isomer impurities, namely the preparation method of a compound I, the preparation method of a compound II, the preparation method of a compound III and the preparation method of a compound IV, and is characterized in that the preparation method of the compound I comprises the following steps:

s1, reacting tenofovir and triphenyl phosphite under the action of a catalyst under an alkaline condition to generate monophenyl tenofovir;

s2, generating phosphoryl chloride monophenyl tenofovir under the action of a chlorinating reagent;

s3, dissociating D-alanine isopropyl ester hydrochloride under the alkaline condition to obtain D-alanine isopropyl ester;

s4, condensing the phosphoryl chloride monophenyl tenofovir and the D-isopropyl alanine to generate a crude compound I;

s5, resolving the crude product of the compound I by adopting L-tartaric acid to obtain a compound I;

wherein said compound I is the code for the impurity 9- { (R) -2- [ ((R) - { [ (R) -1- (isopropoxycarbonyl) ethyl ] amino } -phenoxyphosphoryl) methoxy ] propyl } adenine, and crude compound I is a mixture of compound I and its diastereoisomers.

2. The method for preparing the impurity of the isomer of propane fumarate tenofovir according to claim 1, wherein the alkali used in the basic condition of S1 is one or more selected from triethylamine, sodium hydroxide and potassium hydroxide, and the catalyst in S1 is DMAP; the chlorinating agent in S2 is selected from one or more of thionyl chloride, chlorine and NCS; the alkali used in the alkaline condition in S3 is one or more selected from potassium carbonate, sodium carbonate, potassium bicarbonate and sodium bicarbonate.

3. The method for preparing the isomer impurity of propane fumarate tenofovir according to claim 1, wherein the method for preparing the compound II comprises the following steps:

s1, generating phosphoryl chloride monophenyl tenofovir under the action of a chlorinating reagent;

s2, dissociating L-alanine isopropyl ester hydrochloride under the alkaline condition to obtain L-alanine isopropyl ester;

s3, condensing the phosphoryl chloride monophenyl tenofovir and the L-isopropyl alanine to generate a compound II crude product;

s4, resolving the crude compound II by using L-tartaric acid to obtain a compound II;

wherein said compound II is the code for the impurity 9- { (R) -2- [ ((R) - { [ (S) -1- (isopropoxycarbonyl) ethyl ] amino } -phenoxyphosphoryl) methoxy ] propyl } adenine and crude compound II is a mixture of compound II and its diastereoisomers.

4. The method for preparing the isomer impurity of propofol fumarate and tenofovir according to claim 3, wherein, the chlorinating agent in S1 is selected from one or more of thionyl chloride, chlorine, NCS; the alkali used in the alkaline condition in S2 is one or more selected from potassium carbonate, sodium carbonate, potassium bicarbonate and sodium bicarbonate.

5. The method for preparing the isomer impurity of propane fumarate tenofovir according to claim 1, wherein the preparation method of the compound III comprises the following steps:

s1, generating phosphoryl chloride monophenyl tenofovir under the action of a chlorinating reagent;

s2, dissociating D-alanine isopropyl ester hydrochloride under the alkaline condition to obtain D-alanine isopropyl ester;

s3, condensing the phosphoryl chloride monophenyl tenofovir and the D-isopropyl alanine to generate a compound III crude product;

s4, recrystallizing the compound III to obtain a compound III;

wherein said compound III is the code for the impurity 9- { (R) -2- [ ((S) - { [ (R) -1- (isopropoxycarbonyl) ethyl ] amino } -phenoxyphosphoryl) methoxy ] propyl } adenine and crude compound III is a mixture of compound III and its diastereoisomers.

6. The method for preparing the isomer impurity of propofol fumarate and tenofovir according to claim 5, wherein, the chlorinating agent in S1 is selected from one or more of thionyl chloride, chlorine, NCS; the solvent used for the recrystallization in S4 is a mixed solvent of toluene and acetonitrile.

7. The method for preparing the isomer impurity of malonic acid propofol, tenofovir fumarate as claimed in claim 6, wherein the volume ratio of toluene to acetonitrile in the mixed solvent is 1:3-1: 5.

8. The method for preparing an impurity, namely, propiofuran fumarate and tenofovir disoproxil according to claim 1, wherein the method for preparing the compound IV comprises the following steps:

s1, generating a single-phenyl tenofovir isomer by the action of a catalyst under the alkaline condition of the tenofovir isomer and triphenyl phosphite;

s2, generating phosphoryl chloride monophenyl tenofovir isomer under the action of a chlorinating reagent;

s3, dissociating D-alanine isopropyl ester hydrochloride under the alkaline condition to obtain D-alanine isopropyl ester;

s4, condensing the phosphoryl chloride monophenyl tenofovir isomer and the D-isopropyl alanine to generate a compound IV crude product;

s5, resolving the crude compound IV by using D-tartaric acid to obtain a compound IV;

wherein said compound IV is the code for the impurity 9- { (S) -2- [ ((R) - { [ (R) -1- (isopropoxycarbonyl) ethyl ] amino } -phenoxyphosphoryl) methoxy ] propyl } adenine and the crude compound IV is a mixture of compound IV and its diastereoisomers.

9. The method for preparing the impurity of the isomer of propane fumarate tenofovir according to claim 8, wherein the base used in the basic condition of S1 is one or more selected from triethylamine, sodium hydroxide and potassium hydroxide; the chlorinating agent in S2 is selected from one or more of thionyl chloride, chlorine and NCS; the alkali used in the alkaline condition in S3 is one or more selected from potassium carbonate, sodium carbonate, potassium bicarbonate and sodium bicarbonate.

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