KR20210125298A - New process for the preparation of Tenofovir alafenamide hemi-tartrate - Google Patents

Abstract

The present invention relates to a novel method for manufacturing tenofovir alafenamide hemitartrate. The manufacturing method according to the present invention can produce crystals of tenofovir alafenamide hemitartrate with high yields and good purity in a simple process.

Description

New process for the preparation of Tenofovir alafenamide hemi-tartrate

The present invention relates to a novel process for the preparation of tenofovir alafenamide hemitartrate.

Tenofovir (Formula: (R)-9-(2-phosphonomethoxypropyl)adenine, PMPA) represented by the following Chemical Formula A is a nucleotide reverse transcriptase inhibitor that can effectively treat infections of various viruses, at physiological pH. It exhibits low cell membrane permeability, poor oral bioavailability and certain toxicity due to the divalent anion of the phosphate salt formed.

(Formula A)

In order to compensate for the above disadvantages of tenofovir, tenofovir disoproxil fumarate represented by Formula B, which is a diester prodrug of PMPA, was developed by Gilead.

(Formula B)

However, tenofovir disoproxil fumarate cannot maintain an appropriate concentration in the infected area, and it not only poses a specific renal toxicity risk, but also has safety issues such as a decrease in bone density. Accordingly, tenofovir alafenamide free base represented by the following formula (C), which is a prodrug containing tenofovir as a main ingredient, such as tenofovir disoproxil fumarate from Gilead (Formula: 9-[(R)-2) -[[(S)-[[(S)-1-(isopropoxylcarbonyl)-1-methyl]ethyl]amino]phenoxyphosphinyl]methoxyl]propyl]adenine) GS7340 was developed (see WO2013052094) ).

The tenofovir alafenamide is converted into tenofovir diphosphate in the body and integrated into viral DNA by HBV reverse transcriptase, thereby inhibiting HBV proliferation and causing a DNA chain termination reaction.

(Formula C)

On the other hand, tenofovir alafenamide free base is not suitable for the preparation and dissolution of pharmaceutical preparations due to its low solid melting point and low solubility in water. Tenofovir alafenamide monofumarate, which is a salt form of tenofovir alafenamide with excellent physical properties, has been developed (see US Patent No. 7,803,788).

The tenofovir alafenamide fumarate exhibits similar antiviral efficacy even at a 25 mg dose, which is 1/10 less than that of the existing tenofovir disoproxil fumarate 300 mg, and reduces toxicity to kidney and bone metabolism It is an improved tenofovir drug. Tenofovir alafenamide fumarate reduced the systemic exposure of the drug by lowering the plasma tenofovir concentration by 89% compared to tenofovir disoproxil fumarate. Keep similar to tenofovir disoproxil fumarate.

However, the tenofovir alafenamide fumarate has poor chemical stability and thermodynamic stability, so it is difficult to easily separate it from impurities of (R)-9-(2-phosphonomethoxypropyl)adenine.

Accordingly, tenofovir alafenamide hemifumarate represented by the following formula (D) having the advantage of being easily separated from the above impurities compared to tenofovir alafenamide monofumarate (Formula: 9-[(R) Developed by -2-[[(S)-[[(S)-1-(isopropoxycarbonyl)-1-methyl]ethyl]amino]phenoxyphosphinyl]methoxyl]propyl]adenine hemifumarate) (refer to Korean Patent No. 10-1612642).

(Formula D)

As described above, tenofovir alafenamide hemifumarate can be easily separated from impurities of (R)-9-(2-phosphonomethoxypropyl)adenine, but such a salt of tenofovir alafenamide hemifumarate The preparation method is complicated in that it is necessary to add tenofovir alafenamide hemifumarate seed along with fumaric acid and tenofovir alafenamide during the preparation process.

Accordingly, as it is required to develop a new salt that is simple to prepare and has high physical properties and chemical stability, tenofovir alafenamide hemitartrate represented by the following formula E, in which some crystalline forms have high biological activity, has been developed, the tenofovir alafenamide hemitartrate Beer alafenamide hemitartrate has chemical stability and thermal stability suitable for use as a drug compared with tenofovir alafenamide fumarate (see Chinese Patent No. 105237571 B, etc.).

(Formula E)

값 및 그 상세한 값 또는 도면에 대한 언급도 없다.Regarding such tenofovir alafenamide hemitartrate, Chinese Patent Registration No. 105237571 B describes tenofovir alafenamide hemitartrate and a method for preparing tenofovir alafenamide hemisuccinate. However, in the preparation of tenofovir alafenamide hemitartrate and hemisuccinate, n-hexane, which is prohibited for use in pharmaceuticals, and petroleum ether, which does not yet have a residual solvent standard on the ICH Guideline, are used, and it is preferable from a pharmaceutical standpoint. can't In addition, in the above patent, 2 of XRD crystal analysis for crystalline forms

No reference is made to the values and their detailed values or drawings.

값 및 그 상세한 도면을 개시하고 있다.Chinese Patent Publication No. 104926872 A discloses a method for preparing tenofovir alafenamide hemitartrate and a value of DSC crystallization analysis for the prepared crystalline form and detailed drawings thereof, XRD crystallization analysis 2

Values and detailed drawings thereof are disclosed.

Indian Patent Publication No. 04260/CHE/2013 discloses various salts such as succinate, L-tartrate, and methanesulfonate of tenofovir alafenamide. In particular, Indian Patent Publication No. 00118/MUM/2014 In the heading, L-tartaric acid, D-tartaric acid, (1S)-(+)-10-camphorsulfonic acid, (1R)-(-)-10-camphorsulfonic acid, camphoric acid, L-only of tenofovir alafenamide Delic acid, D-mandelic acid, benzoic acid. A method for purifying the optical isomers of tenofovir alafenamide by salt formation using L-malic acid, succinic acid, oxalic acid, orthophosphoric acid and maleic acid followed by desalting is mentioned.

The present invention relates to a novel manufacturing method different from the manufacturing methods of the above patents, which are the prior art. It has excellent chemical and thermal stability, no hygroscopicity, and high yield of tenofovir alafenamide hemitartrate crystals useful in pharmaceutical manufacturing. and to provide a method capable of easily manufacturing a large quantity with high purity.

International Patent Publication WO2013052094 US Patent No. 7,803,788 Domestic registered patent 10-1612642 China Registered Patent No. 104926872 China Registered Patent No. 105237571 Indian Patent Publication No. 04260/CHE/2013 Indian Patent Publication No. 00118/MUM/2014

Hemitartrate of tenofovir alafenamide prepared from the free base of existing tenofovir alafenamide is of very low purity. In general, when a salt is prepared, the purity is increased, but in the case of tenofovir alafenamide, there is no change in the purity or the purity is sometimes low. This is because (R)-9-(2-phosphonomethoxypropyl)adenine produced by decomposition of tenofovir alafenamide upon dissolution is not removed.

Therefore, while the present inventors were conducting research to remove (R)-9-(2-phosphonomethoxypropyl)adenine, (R)-9-(2-phosphonomethoxypropyl)adenine was removed from the organic solvent. It was confirmed that it was difficult and easy to remove from water, and when the salt was prepared after removal, it was confirmed that the content of (R)-9-(2-phosphonomethoxypropyl)adenine was very low. Based on this study, we developed an effective method for industrial mass production by optimizing the production method of tenofovir alafenamide hemitartrate and the crystallization conditions to obtain crystals with excellent chemical and thermal stability and no hygroscopicity. invention was completed.

An object of the present invention is to provide a method for easily producing tenofovir alafenamide hemitartrate with high yield and high purity, which is pharmaceutically stable and has a form and physical properties useful as a pharmaceutical.

Another object of the present invention is to provide a crystalline form of tenofovir alafenamide hemitartrate prepared by the above preparation method.

The method for preparing the crystal form of tenofovir alafenamide hemitartrate according to the present invention comprises the steps of: A) dissolving or suspending tenofovir alafenamide free base, or tenofovir alafenamide organic acid or inorganic acid salt in an organic solvent Wow; B) adding a solution of an inorganic base in water; C) concentrating the organic layer after dehydration; D) adding and dissolving alcohol and an organic solvent, or a mixture thereof to the residue obtained by the concentration, and then adding L-tartaric acid; E) reacting the reaction mixture through stirring, and aging the crystals produced by the reaction through cooling; will be manufactured with

The method for producing tenofovir alafenamide hemitartrate crystals according to the present invention is easy to manufacture, has a high purity, has a high yield, and has the effect of lowering the production cost, and has an excellent form for use as a drug, such as high stability and physical properties.

1 is an X-ray diffraction spectrum of a crystalline form of tenofovir alafenamide hemitartrate according to Example 1. FIG.
FIG. 2 is a differential scanning calorimetry spectrum for the crystalline form of tenofovir alafenamide hemitartrate according to Example 1. FIG.
3 is an X-ray diffraction spectrum of the crystalline form of tenofovir alafenamide hemitartrate according to Example 2. FIG.
4 is a differential scanning calorimetry spectrum for the crystalline form of tenofovir alafenamide hemitartrate according to Example 2. FIG.

Hereinafter, preferred embodiments and the like will be described to help the understanding of the present invention. However, the embodiments according to the present invention may be modified in various other forms, and the scope of the present invention is not limited to the scope of the present invention due to the following examples.

The method for preparing the crystal form of tenofovir alafenamide hemitartrate according to the present invention comprises the steps of: A) dissolving or suspending tenofovir alafenamide free base, or tenofovir alafenamide organic acid or inorganic acid salt in an organic solvent Wow; B) adding a solution of an inorganic base in water; C) concentrating the organic layer after dehydration; D) adding and dissolving alcohol and an organic solvent, or a mixture thereof to the residue obtained by the concentration, and then adding L-tartaric acid; E) reacting the reaction mixture through stirring, and aging the crystals produced by the reaction through cooling;

Specifically, in step A), the tenofovir alafenamide organic acid or inorganic acid salt is fumaric acid, ferulic acid, phosphoric acid, succinic acid, citric acid, L-tartaric acid, D-tartaric acid, methanesulfonic acid, (1S)-(+)- 10-camphorsulfonic acid, (1R)-(-)-10-camphorsulfonic acid, camphoric acid, L-mandelic acid, D-mandelic acid, lactic acid, benzoic acid. Tenofovir alafenamide organic acid using L-malic acid, maleic acid, hydrochloric acid, benzenesulfonic acid, ethanesulfonic acid, galactaric acid, malonic acid, pamophosphoric acid, sebacic acid, vanillic acid, oxalic acid, sulfuric acid, orthophosphoric acid or maleic acid or an inorganic acid salt.

In addition, as the organic solvent as the reaction solvent in step A), ethyl acetate, methylene chloride, chloroform, isopropyl ether, methyl tertbutyl ether or a mixture thereof is used, and methylene chloride or ethyl acetate is preferably used.

In step B), the inorganic base is sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, or sodium hydrogen carbonate, preferably sodium hydrogen carbonate. The concentration of the aqueous inorganic base solution is 0.1% to 20%, and the amount of the aqueous inorganic base solution is 0.1 to 20 times compared to 1 g of tenofovir alafenamide free base, or tenofovir alafenamide organic acid or inorganic acid salt.

As the solvent used in step D), methanol, ethanol, isopropanol, n-butanol and ethyl acetate or mixtures thereof are used, and preferably a mixture of isopropanol and ethyl acetate is used.

The molar ratio of L-tartaric acid to tenofovir alafenamide free base or tenofovir alafenamide organic acid or inorganic acid salt is 1:0.45 to 1:0.60, preferably tenofovir alafenamide free base. , or tenofovir alafenamide organic acid or inorganic acid salt and L-tartaric acid are used in a molar ratio of 1:0.50 to 1:0.55.

In step E), the reaction mixture solution of step D is stirred at a temperature of 20-100° C. for 0.5 to 12 hours, preferably for 0.5 to 2 hours, to obtain tenofovir alafenamide free base, or tenofovir alafen. The amide organic or inorganic acid salt is reacted with L-tartaric acid.

After the reaction, the reaction mixture is first aged by stirring for 0.5 to 24 hours, preferably 1 to 3 hours at a temperature of 20 to 30° C., and then again at a temperature of 0 to 15° C. for 0.5 to 24 hours, preferably 1 After secondary aging by stirring for ~4 hours or the like, the precipitated crystals are filtered to obtain crystals of tenofovir alafenamide hemitartrate.

Comparative Examples and Examples are disclosed below.

<Comparative Example 1>

According to the method of Example 8 described in Chinese Patent Registration No. 104926872, tenofovir alafenamide hemitartrate was prepared (yield: 93.1%, purity 99.71%)

<Example 1>

Preparation of tenofovir alafenamide hemitartrate from tenofovir alafenamide free base

After dissolving 20.0 g of tenofovir alafenamide free base in 100 ml of methylene chloride, 20 ml of a 5% aqueous sodium hydrogen carbonate solution was added and stirred. The organic layer is separated, dehydrated, and then concentrated. After the concentrate and 3.15 g of L-tartaric acid were suspended in 120 ml of isopropanol and 80 ml of ethyl acetate, the reaction was carried out by stirring at 80° C. for 30 minutes. After cooling to room temperature and stirring for 1 hour, the precipitated crystals were aged by stirring at 0-5°C for 2 hours, filtered, washed with ethyl acetate, and dried under reduced pressure at 40°C to obtain the crystalline form of the title compound. (Yield: 94.9%, Purity: 99.87%)

The XRD pattern of the crystal obtained in Example 1 is shown in Fig. 1, and Fig. 2 shows the DSC of the crystal.

<Example 2>

Preparation of tenofovir alafenamide hemitartrate from tenofovir alafenamide organic acid salt

20.0 g of tenofovir alafenamide fumarate salt was suspended in 100 ml of ethyl acetate, and 60 ml of 5% aqueous sodium hydroxide solution was added thereto and stirred. The organic layer is separated, dehydrated, and then concentrated. The concentrate and 2.61 g of L-tartaric acid were suspended in 120 ml of n-butanol and 80 ml of ethyl acetate, stirred at 85° C. for 30 minutes to proceed with the reaction, then slowly cooled to room temperature, stirred for 1 hour, and stirred at 0-5° C. for 2 hours The precipitated crystals were aged, filtered, washed with ethyl acetate, and dried under reduced pressure at 40°C to obtain crystals of the title compound having the same crystalline form as those of Example 1 (yield: 93.9%, purity: 99.88%).

<Crystalline Analysis>

go. Powder X-ray diffraction (XRD) analysis of tenofovir alafenamide hemitartrate crystalline form of Example 1

회절각이 4.16±0.2°, 9.55±0.2°, 17.89±0.2°, 19.07±0.2°, 19.84±0.2°, 20.50±0.2°및 21.80 ± 0.2°에서 각각 피크를 보였다.As a result of powder X-ray diffraction analysis on the crystal form of tenofovir alafenamide hemitartrate obtained in Example 1, 2

The diffraction angles showed peaks at 4.16±0.2°, 9.55±0.2°, 17.89±0.2°, 19.07±0.2°, 19.84±0.2°, 20.50±0.2° and 21.80±0.2°, respectively.

At this time, the measurement conditions of the powder X-ray diffraction (XRD) spectrum are as follows.

1) Apparatus: Rigaku's MiniFlex 600/X-ray source: Cu

2) Tube voltage: 40 kV / Tube current: 15mA

3) Divergence slit: 1°/scatter slit: 1°/receive slit: 0.15mm

/샘플링 간격 0.04℃4) Injection range: 3 to 40° 2

/sampling interval 0.04℃

5) Scan speed: 10°/min

me. Differential scanning calorimetry (DSC) analysis

Differential scanning calorimetry (DSC) analysis was performed on the crystal form of tenofovir alafenamide hemitartrate obtained in Example 1, and the results are attached to FIG. 2 . As shown in FIG. 2 , the crystal showed an onset temperature of 174±1°C and an endothermic peak of 176±1°C.

The measurement conditions of the differential scanning calorimeter spectrum are as follows.

1) Device: Q20 (TA instrument)

2) Measuring range: 10 to 200°C/temperature increase interval: 10°C/min

all. Powder X-ray diffraction (XRD) analysis of tenofovir alafenamide hemitartrate crystalline form of Example 2

회절각이 4.16±0.2°, 9.55±0.2°, 17.89±0.2°, 19.07±0.2°, 19.84±0.2°, 20.50±0.2°및 21.80±0.2°에서 각각 피크를 보였다.As a result of performing powder X-ray diffraction analysis on the crystal form of tenofovir alafenamide hemitartrate obtained in Example 2, 2

The diffraction angles showed peaks at 4.16±0.2°, 9.55±0.2°, 17.89±0.2°, 19.07±0.2°, 19.84±0.2°, 20.50±0.2° and 21.80±0.2°, respectively.

At this time, the measurement conditions of the powder X-ray diffraction (XRD) spectrum are as follows.

1) Apparatus: Rigaku's MiniFlex 600/X-ray source: Cu

2) Tube voltage: 40 kV / Tube current: 15mA

3) Divergence slit: 1°/scatter slit: 1°/receive slit: 0.15mm

/샘플링 간격 0.04℃ 4) Injection range: 3 to 40° 2

/sampling interval 0.04℃

5) Scan speed: 10°/min

La. Differential scanning calorimetry (DSC) analysis

Differential scanning calorimetry (DSC) analysis was performed on the crystal form of tenofovir alafenamide hemitartrate obtained in Example 2, and the results are attached to FIG. 4 . As shown in FIG. 4 , the crystal showed an onset temperature of 174 + 1 °C and an endothermic peak of 176 + 1 °C, similar to the crystal form of Example 1.

The measurement conditions of the differential scanning calorimeter spectrum are as follows.

1) Device: Q20 (TA instrument)

2) Measuring range: 10 to 200°C/temperature increase interval: 10°C/min

mind. Hygroscopicity test

The hygroscopicity of the tenofovir alafenamide hemitartrate crystals obtained in Examples 1 and 2 and the crystals obtained in Comparative Example 1 was measured, and the results are compared and shown in Table 1 below. Specific hygroscopicity measurement conditions are measurements of the water content (K.F. moisture %) of each compound over time under the conditions of a temperature of 25° C. and a relative humidity of 30%.

Sample Early 1 week 2 weeks 4 weeks Example 1
(tenofovir alafenamide hemitartrate)
0.12%
0.12%
0.12%
0.12% Example 2
(tenofovir alafenamide hemitartrate)
0.14%
0.14%
0.14%
0.14% Comparative Example 1
(tenofovir alafenamide hemitartrate)
0.16%
0.16%
0.18%
0.19%

As shown in Table 1, tenofovir alafenamide hemitartrate prepared by the preparation methods of Examples 1 and 2 of the present invention did not exhibit hygroscopicity. That is, compared with the initial water content, the water content is maintained at an almost equal level even after 1 week, 2 weeks, or 4 weeks. On the other hand, as the tenofovir alafenamide hemitartrate of Comparative Example 1 elapsed, it was confirmed that the moisture content in the air increased as the time elapsed.

bar. Stability test

The results of HPLC analysis of the tenofovir alafenamide hemitartrate crystals of Examples 1 and 2 of the present invention and the tenofovir alafenamide hemitartrate crystals prepared in Comparative Example when stored at room temperature were analyzed by HPLC. It is shown in Table 2 below.

Sample Early 1 week 2 weeks 4 weeks Example 1
(tenofovir alafenamide hemitartrate)
99.874%
99.873%
99.874%
99.873% Example 2
(tenofovir alafenamide hemitartrate)
99.881%
99.881%
99.881%
99.880% Comparative Example 1
(tenofovir alafenamide hemitartrate)
99.711%
99.709%
99.706%
99.704%

As can be seen in Table 2 above, the crystals of tenofovir alafenamide hemitartrate of Examples 1 and 2 of the present invention showed little difference in HPLC analysis results even after a period of time when stored at room temperature, whereas Comparative Example 1 of tenofovir alafenamide hemitartrate crystals were stored at room temperature, and HPLC analysis results gradually decreased over time. It can be seen that the crystal of tenofovir alafenamide hemitartrate of Example 1 is very stable.

As described above, the tenofovir alafenamide hemitartrate of the present invention is easy to store and has excellent storage stability, and this stability is also shown to be consistent with the results of the above hygroscopicity test.

Claims (8)

A process for preparing the crystalline form of tenofovir alafenamide hemitartrate, comprising the steps of: A) dissolving or suspending tenofovir alafenamide free base or tenofovir alafenamide organic acid or inorganic acid salt in an organic solvent; B) adding a solution of an inorganic base in water; C) concentrating the organic layer after dehydration; D) adding and dissolving alcohol and an organic solvent, or a mixture thereof to the residue obtained by the concentration, and then adding L-tartaric acid; E) reacting the reaction mixture through stirring, and aging the crystals produced by the reaction through cooling; According to claim 1, wherein said tenofovir alafenamide organic acid or inorganic acid salt in step A) is fumaric acid, ferulic acid, phosphoric acid, succinic acid, citric acid, L-tartaric acid, D-tartaric acid, methanesulfonic acid, (1S)-( +)-10-camphorsulfonic acid, (1R)-(-)-10-camphorsulfonic acid, camphoric acid, L-mandelic acid, D-mandelic acid, lactic acid, benzoic acid. L-malic acid, maleic acid, hydrochloric acid, benzenesulfonic acid, ethanesulfonic acid, galactaric acid, malonic acid, pamophosphoric acid, sebacic acid, vanillic acid, oxalic acid, sulfuric acid, orthophosphoric acid, characterized in that the salt of maleic acid, tenofo A method for preparing a crystalline form of beer alafenamide hemitartrate. The method according to claim 1, wherein the crystallization solvent is methanol, ethanol, isopropanol, n-butanol and ethyl acetate or mixtures thereof. 4. The method according to claim 3, wherein the crystallization solvent is a mixture of isopropanol and ethyl acetate. According to claim 1, wherein the inorganic base is sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, or sodium hydrogen carbonate, the concentration of the inorganic base aqueous solution is 0.1% to 20%, the amount of the inorganic base aqueous solution used is tenofovir ala 0.1 to 1 g of fenamide free base, or tenofovir alafenamide organic or inorganic acid salt A method for producing a crystal form of tenofovir alafenamide hemitartrate, characterized in that the ratio is 20 times. A crystalline form of tenofovir alafenamide hemitartrate, prepared by the method according to any one of claims 1 to 5 7. The method of claim 6, wherein in powder X-ray diffraction (XRD) analysis 2

Tenofovir alafenamide hemitart with diffraction angles of 4.16±0.2°, 9.55±0.2°, 17.89±0.2°, 19.07±0.2°, 19.84±0.2°, 20.50±0.2° and 21.80±0.2° rate crystalline form The crystalline form of claim 6, wherein tenofovir alafenamide hemitartrate has an onset temperature of 174±1° C. and an endothermic peak of 176±1° C. in differential scanning calorimetry (DSC) analysis.

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