KR20160135112A - A fast dissolving film-formulation for oral dosage of tenofovir disoproxil fumarate and the manufacturing method thereof - Google Patents

Abstract

The present invention relates to an oral dissolving film dosage form using tenofovir disoproxil fumarate (Teno-DF), which is used as a therapeutic agent for hepatitis B or AIDS due to physiological activity promoting function thereof, and to a preparing method therefor. The method for preparing tenofovir disoproxil fumarate of the present invention can increase the yield and purity in the synthesis of the compound by using an ion exchange resin (Dowex 50W hydrogen form, sulfonic acidic cation exchange resin), and furthermore, unlike a dosage form obtained by coating a film on a tablet as the conventional art, the oral dissolving film dosage form provided in the present invention is convenient for a patient to take.

Description

[0001] The present invention relates to an oral dissolution film (ODF) formulation using tenofovir disoproxil fumarate and a manufacturing method thereof, and a fast dissolving film-

The present invention relates to a method for effectively synthesizing the above-mentioned compounds by using an appropriate catalyst and the use of an ion exchange resin in the synthesis of tenofovir and its derivatives used for hepatitis B and HIV infection due to antiviral function And a method for producing a therapeutic agent for hepatitis B infection or HIV infection, which comprises using the above-described synthetic method and high-purity ginsenosides, as an effective ingredient, into a formulation of an oral dissolution film.

Up to now, no case has been reported for the synthesis of tenofovir disoproxyl fumarate in the same manner as the present invention. Particularly, in the synthesis of tenofovir, the hydrolysis process included in the intermediate process and the alkali neutralization process proceeded after the hydrolysis are included, so that many by-products and decomposition products are produced due to the above processes and mixed with such by- The tenofovir has low solubility in water and a low synthesis yield due to a problem of containing crystal water, and therefore, when proceeding to the next step reaction of the produced taboflavir material, it is necessary to remove the crystal water contained in the tenofovir molecule The occurrence of various types of ester compounds as by-products can be reduced. Therefore, in order to remove the crystal water from Tenofovir H ₂ O (MW 305.3) containing the crystalline water produced above, it generally includes a step of drying under reduced pressure at a high temperature. However, in the synthesis method of the present invention, It is possible to minimize the byproducts generated in the synthesis step of the synthesis solution so that the synthesis solution is precisely neutralized using an ion exchange resin and the treatment with an organic solvent is relatively There is an advantage that it is possible to synthesize high purity Tenofovir disiproxyl fumarate having a small amount of by-products and decomposition products by passing through a simple decompression drying step proceeding at a low temperature.

 The present invention also relates to a pharmaceutical composition for the treatment of hepatitis B or AIDS, which comprises, as an active ingredient, GS-10, which is synthesized by the synthesis method of the present invention, To a method of making oral dissolution film formulations.

Viread®, a hepatitis B treatment developed by the Gillery Advisory Company of the United States, has the generic name Tenofovir disoproxil fumarate (hereinafter referred to as 'Teno-DF', MP 219 ° C, MW 635.52) and the chemical name according to the structural formula is 9- [2- (R) - [[bis [[(isopropoxycarbonyl) oxy] methoxy] phosphinoyl] methoxy] propyl] adenine fumaric acid (9 - [(R) -2 - [[bis [[(isopropoxycarbonyl) oxy] methoxy] phosphinyl]] methoxy] propyl] adenine fumarate.

 The Teno-DF has been approved by the US FDA for the treatment of AIDS in 2001. In April 2008, Teno-DF was approved as a hepatitis B (HBV) It was once again approved as a hepatitis treatment. Teno-DF was prepared by replacing the isopropylcarbonylmethoxy group, a structural component of the lipid component, with tenofovir to enhance the in vivo absorption capacity of Tenofovir (MP 267-280 ° C, MW 287.21) as a prodrug . Therefore, the most important problem in the synthesis of the Teno-DF is to synthesize the main component having the antiviral effect with high yield and high purity.

 (R) -9- [2- (hydroxy) propyl] adenine, MW. 193.2). The raw material used in the synthesis of tenofovir is hydroxypropyladenine (HPA, Minimizing impurities and degradation materials is an important challenge.

Tenofovir (phosphonomethoxypropyl adenine,

(R) -9- [2- (phosphonomethoxy) propyl] adenine, PMPA), which is one of the processes for synthesizing the HPA raw material, - (Diethylphosphono methoxy) propyl] adenine, the yield and purity of the final product, Tenofovir, depends on the catalytic chemical reaction method. However, since the process of synthesizing DEPMPA from HPA at a high yield and hydrolyzing it to produce highly pure Tenofovir (PMPA) is very difficult, a great deal of research has been conducted and a synthesis method emphasizing excellent And is continuously applying for papers and patents.

 The catalytic chemical reaction is carried out by using various alkaline catalysts, and the reaction organic solvent and the extraction method are differently used according to each reaction. As the alkoxide catalyst as a condensation catalyst, methylene phosphate monomer is introduced in the presence of lithium butoxide, a catalyst such as magnesium butoxide, magnesium isopropoxide, sodium amide and magnesium acetate, and butylmagnesium chloride in advance The raw material, diethyl para-toluenesulfonyloxymethylphosphonate

(Diethyl-p-toluenesulfonyloxymethylphosphonate, DEPTSMP) and HPA. Once the Tenofovir is produced in high purity and high yield by using the above-mentioned catalyst or the like, Teno-DF, which is a precursor derivative due to the subsequent reaction, can be produced according to the conventional method.

 The problem that occurs when a large amount of impurities are present in the synthesized Tenofovir is that the stability of the product using the final product, Teno-DF, is lowered due to a large influence on the reaction of the next step for synthesizing Teno-DF, The decomposition products which are not filtered during the separation and removal of by-product residues in the reaction process are generated because the decomposition products exist in excess, exceeding the allowable standards, and industrialization using Teno-DF becomes very difficult. For this reason, the price of the raw material is considerably high.

Tenofovir disoproxil (Teno-D, MP 61-65 ° C, MW 519.45) has been reported to produce decomposed products in storage and produce a self-polymer, Teno-D Dimer, if the purity is poor (Organic Process Research & Development, 2010, Vol. 14, 1194-1201, Pharmaceutical Research, 2001, Vol. 18, 234-237).

≪ Synthesis Reaction Scheme of Tenofovirdisofroxyl fumarate (Teno-DF) & Process Diagram >

<Review of existing patent documents>

 0008-1. Published Patent CN 102219805A (Mar. 10, 2011) Novel production process of Tenofovir.

In the above patent, HPA is methylene phosphorylated using various catalysts, and the resulting diethyl ester substance is hydrolyzed. Then, the strongly acidic substance is neutralized to pH 3, which is a hydrogen ion concentration, by using 40% sodium hydroxide, And to a method for obtaining crystals. However, in the above patent, in consideration of the solubility of tenofovir, it is inevitable to use a 40% sodium hydroxide solution, which is a concentrated alkaline neutralizing solution. In this method, it is difficult to obtain tenofovir, which is a substance having an accurate isoelectric point.

 0008-2. Open Patent CN 102295660A (July 4, 2011) Process for synthesizing Tenofovir

This patent relates to a process for preparing ethyl phosphate from HPA and then hydrolyzing it with hydrobromic acid followed by adjusting the pH to about 3.2 with sodium hydroxide solution.

 0008-3. Open Patent CN 103374039A (Apr. 20, 2012) Synthesis of Tenofovir

The above patent relates to a method for obtaining PH by adjusting the pH to 2.5 to 4 after hydrolysis by the same process as the above-mentioned 0008-2.

 0008-4. [Patent Document 1] Japanese Patent Application Laid-Open No. 102899367B (2012.09.24) A biological method and chemically combined with the method of synthetic Tenofovir

The above patent relates to a method for neutralization by neutralizing PH to 3.0 using a carbonate as a neutralized alkali solution.

 0008-5. Registered Patent CN 01239189B (Mar. 17, 2008) Tenofovir, Adefovir and intelligent polymer conjugates and preparation and use thereof.

The above patent relates to a method for adjusting pH to 2 to 3 by using a phosphoric acid buffer solution at neutralization.

 0008-6. Open Patent US 2013/0165413 A1 (2013.06.27) Process for the preparation of Tenofovir

The above patent discloses that various metal alkoxide catalysts are used as alkali condensation catalysts. Various catalysts such as sodium amide and magnesium chloride complex catalysts are used. After the hydrolysis, the reaction solution is concentrated at a high concentration And the pH is neutralized to 2 to 3 using an aqueous alkaline solution. The X-ray powder diffraction pattern (XRPD) of Teno-DF is also presented.

 0008-7. US 2014/0303368 A1 (October 9, 2014) Process for the preparation of Tenofovir

The above patent is similar to the patent of 0008-6, but it relates to a method of synthesizing aluminum and an amic acid catalyst in the synthesis of diethylenepoiovir and then hydrolyzing and then neutralizing PH to 2 to 3 with an aqueous alkaline solution .

0008-8. Open Patent CN 101948485B (Jul. 25, 2012) Alpha crystal form of tenofovir disoproxil fumarate, and preparation method and application thereof

The above patent relates to a new crystal form of tenofovir fumarate, which is an effective ingredient for producing a pharmaceutical composition, and the novel crystalline form of tenofovir fumarate described in the above patent is characterized by excellent solubility and human hygroscopicity. In addition, the above-mentioned patents disclose that the above-mentioned tenofovir fumarate crystal form can be produced by a formulation such as an oral tablet, a controlled release tablet, a film-coated tablet and the like, which is coated with a film using the above-mentioned tenofovir fumarate crystal form (Publication No. 32 of the publication), so that it is possible to prepare tablets for oral dissolution using the same, and the contents of the film formulation for oral dissolution, which is another kind of medicines, are described It is not.

 <Non-Patent Documents>

 0008-9. PMP (PMPA, (R) -9- [2- (Phosphononomethoxy) propyl] adenine), &lt; / RTI &gt; Tetrahedron Letters 39 (1998) 1853-1856)

In the above document, the diester compound produced after the methylene phosphorylation reaction is carried out by using sodium hydride (NaH) or lithium tertiarybutoxide as the catalyst from the HPA, is hydrolyzed with a bromotrimethylsilane solution, and then a 50% aqueous solution of sodium hydroxide To neutralize the PH to 3.

 0008-10. Organic Process Research & Development (2010), Vol.14, 1194p-1201, Process Improvements for the manufacture of Tenofovir disoproxil fumarate at commercial scale.

The above literature discloses that, when methylene phosphorylated ester is prepared by using HPA as a catalyst of magnesium tertiary butoxide and hydrolyzed to obtain Tenofovir crystals, the pH is adjusted to 2.8 to 3.2 with a 40% sodium hydroxide aqueous solution to obtain crystals Lt; / RTI &gt;

 0008-11. Authorized USP Pending Monograph version 1 (2011) Tenofovir Disoproxil Fumarate

The USP Pending specification of the above literature limits the content of 10 or more impurities, thereby standardizing the purity of the raw materials. It can be seen from the above document that the original developer (Gilead, USA) has installed a technical protection network so that other companies can not easily synthesize the tenofovir with precise technology (the expiration period of the original developer Gilead patent is 2017 If the duration of the patent is extended due to delay of the patent examination, the duration of the patent shall be until 2018).

 0008-12. World Health Organization, Final Text for addition to the International Pharmacopoeia (2010.06) -Tenofovir Disoproxil Fumarate

The data in the above literature have been provisionally established as the WHO International Raw Material Standard.

 The technical patents and reference documents of the twelve items described above show that the synthesis yield is not high due to a large amount of impurities generated in the synthesis of tenofovir.

 Accordingly, the present invention relates to a novel synthesis method capable of solving the problems of the technical patents and the references and meeting the high purity and high yield of the end product in the synthesis of tenofovir.

Viread®, a hepatitis B treatment developed by the Gillery Advisory Company of the United States, has the generic name Tenofovir disoproxil fumarate (hereinafter referred to as 'Teno-DF', MP 219 ° C, MW 635.52) and the chemical name according to the structural formula is 9- [2- (R) - [[bis [[(isopropoxycarbonyl) oxy] methoxy] phosphinoyl] methoxy] propyl] adenine fumaric acid salt

(9 - [(R) -2 - [[bis [[(isopropoxycarbonyl) oxy] methoxy] phosphinyl]] methoxy] propyl] adenine fumarate.

 The Teno-DF has been approved by the US FDA for the treatment of AIDS in 2001. In April 2008, Teno-DF was approved as a hepatitis B (HBV) It was once again approved as a hepatitis treatment. Teno-DF was prepared by replacing the isopropylcarbonylmethoxy group, a structural component of the lipid component, with tenofovir to enhance the in vivo absorption capacity of Tenofovir (MP 267-280 ° C, MW 287.21) as a prodrug . Therefore, the most important problem in the synthesis of the Teno-DF is to synthesize the main component having the antiviral effect with high yield and high purity. However, in the conventional technology for synthesizing tenofovir, PH is controlled by using an alkaline substance in the neutralization process, so that many by-products and decomposition products generated during the crystallization of the tenofovir are generated, and the contamination degree of tenofovir And thus the stability of the drug was also problematic in the preparation of the drug using the Tenofovir.

Accordingly, the present invention provides a method for synthesizing tenofovir at a high purity and a high yield by minimizing by-products and decomposition materials generated in the synthesis of tenofovir, which was a problem in the prior art, And to provide a method for preparing the drug into a film formulation to promote its activity.

The present invention relates to a rapid reaction using a catalyst in the synthesis of tenofovir disoproxyl fumarate, a reaction condition under which less impurities and byproducts can be produced, and a method capable of estimating the progress of the reaction conveniently and precisely during the process (Dowex 50W x 4 Hydrogen form, Sulfonic acidic cation exchange resin) during the purification of the intermediate product to obtain the final product, high purity, To a solution film formulation.

The present invention differs from the conventional art in that a synthetic solution is precisely neutralized by mixing an alkaline compound used as a neutralizing agent and an ion exchange resin in the preparation of tenofovir, and then subjected to a simple vacuum drying process , It is possible to synthesize highly pure Tenofovir disoproxyl fumarate with few by-products and decomposition products, and it has an advantage that the stability of the compound is superior to that produced by the conventional preparation method have.

The present invention relates to a process for producing pureer raw materials higher than the synthesis yield of the prior art claiming that they have already been improved in the synthesis of tenofovir, and in particular, isomerization and deliquescence of the reactants occurring during long- And to a method for synthesizing a high-purity raw material by suppressing generation of a molecular polymer during the synthesis process.

 Tenofovir is typically prepared by the reaction of (R) -9- [2- (hydroxy) propyl] adenine (HPA) with diethyl para- toluenesulfonyloxymethylphosphonate (DEPTSMP) It is the main process to obtain the well-being ester compound (DEPMPA) and hydrolyze it again to produce the terbinafil (PMPA), and the resulting terfenovir is the antiviral substance. The synthesis of Teno-D and Teno-DF, which are synthesized later, is merely to make Tenofovir a precursor to physiological activation of lipophilic substances. Until now, as we have seen in several patent cases above, patents on technologies developed globally depend on the improvement of yield and the improvement of purity in this process.

The present invention relates to a process for preparing DEPMPA in HPA by adding calcium hydride to magnesium butoxide as a typical reagent catalyst to shorten the conventional reaction time of 18 hours to within 6 hours, (Dowex 50W, sulfonic acid cation exchage resin) was used to neutralize the ester compound with a reversed alkali solution during the hydrolysis of the terpovorol alcohol, and a precise isoelectric point (PH 3.2 ), Thereby further increasing the synthesis yield of tenofovir. Generally, in the case of PH of 2.5 to 3.5 at neutralization, tenofovir (H ₂ O, single molecule crystal number) is dissolved in the aqueous solution to about 7%, but when neutralized with ion exchange resin, it can be reduced to 4% or less .

 In addition, as a method for closely observing the progress of the reaction during the reaction process, it is inevitable to check the progress of the reaction by the HPLC method which takes a long time in the conventional literature. However, by adding a simple TLC method, the HPLC result which waits for a long time can be taken into consideration in advance.

 Various reagents can be used during the hydrolysis process, but a reagent with few byproducts is selected to be selected as an alkyl salicyl bromide reagent which is fast and can shorten the reaction time. The reaction temperature is set at 75 to 80 ° C for 4 to 6 hours, the hydrolysis reaction is 80 to 90 ° C, and the reaction time is 4 hours to 6 hours. Tenofovir crystallization precipitation process is 0 ~ 5 ℃ in initial reaction.

 <Synthesis of diethylenophorin in HPA Step reaction test>

 (MC) / Ethyl alcohol (EtOH) = 10/2, Silca gel 60 F254, 25 Aluminum sheets 20 X 20 cm, RF (HPA) = 0.3, RF (Diethyl Tenofovir ) = 0.4

 HPLC analysis conditions: USP Pending Monograph Tenofovir Disoproxil Fumarate (2011), UV Detector 260 nm, column 4.6-mm 25 cm; 5micro Packing L1, Col. Temp. 35 C, Sam.Temp. 4 ° C Flow Rate 1 ml / min. Inj. Sz. 10micro L.

 &Lt; Hydrolysis reaction of diethylenepoi &

 Silica gel 60 F254 25 Aluminum Sheets, RF (Tenofovir) = 0.62, Unreacted Tenofovir ester was observed with MC / EtOH = 10/2 as TLC analysis conditions (UV 254 nm): developing solvent methyl alcohol alone solvent (100% MtOH)

 HPLC analysis conditions; Same as during HPA reaction progress test (same as USP Pending M. data)

 <Tenofovir disoproxil (Teno-D) synthesis reaction progress test in Tenofovir>

 TLC analysis conditions (UV 254 nm): developing solvent methyl alcohol single solvent, Silica gel 60 F254, Al. sheets, RF (Teno-D) = 0.75, RF (Tenofovir) = 0.62

 HPLC analysis conditions; Same as in HPA reaction above (same as USP Pending M. data)

 <Comparative Example>

 [Comparative Example 1]

(R) -9- (2- hydroxypropyl) adenine ((R) -9- (2-hydroxypropyl) adenine, HPA)

(R) -propylene carbonate ((R) -propylene carbonate (R)) was obtained by dissolving 40 g (0.296 mol) of adenine in 200 ml of dimethylformamide (DMF) and adding 0.94 g (0.0235 mol, 0.08 equiv.) Of sodium hydroxide (0.390 mol, 1.32 Equiv.) was added thereto, followed by stirring for 30 minutes.

The reaction temperature was raised to 120 DEG C and the mixture was heated and condensed for 24 hours. Thereafter, the reaction was checked by TLC and HPLC. After the reaction was almost completed (90% or more), the reaction temperature was cooled to 70 ° C and 240 ml of isopropyl alcohol (IPA) was added to crystallize. Thereafter, the solution was continuously stirred and cooled. The crystals were filtered at 10 ° C, washed with IPA, and dried in a vacuum dryer at 70 ° C to obtain 41g of HPA (yield: 71.8%).

Synthesis of tenopovir ((R) -9- [2- (phosphonomethoxy) propyl] adenine, Tenofovir)

(Using lithium tertiary butoxide catalyst) (Ref: Tetrahedron Letters 39 (1998) 1853)

40 g (0.21 mol) of HPA was dissolved in 200 ml of dimethylformamide (DMF), and 25.2 g (0.315 mol, 1.5 equiv.) Of lithium t-butoxide was added thereto and suspended. (0.325 mol, 1.5 Equiv.) Of diethyl-p-toluenesulfonyloxy methanephosphonate (DEPTSMP) was added in two portions at a temperature of 30 to 35 DEG C. The mixture was reacted at 80 DEG C for 5 hours or more The reaction progress was confirmed by TLC and HPLC (takes 12 hours).

After the reaction was completed, the reaction mixture was cooled to 20 ° C, and glacial acetic acid was added thereto to adjust pH to 6-6.5. 1,000 ml of ethyl acetate was added thereto, and the mixture was heated to 50 to 60 占 폚, followed by cooling to 40 占 폚 and filtration. The reaction by-product lithium inorganic residue was added to 500 ml of ethyl acetate and stirred. The inorganic matter was removed by filtration at the same temperature. The organic solvents were combined, washed twice with 100 ml of cold distilled water at 10 ° C or less, and washed with 50 ml of saturated sodium chloride solution After the organic layer was dehydrated with anhydrous sodium sulfate, the organic layer was vacuum-distilled to obtain terpovir ester (DEPMPA). 300 ml of toluene was added to the residue (DEPMPA), and the mixture was distilled under reduced pressure to remove DMF and ethyl acetate. The residue was distilled under reduced pressure with 300 ml of cyclohexane. Ethyl acetate and DMF were distilled off and the DEPMPA was dealkylated. Decomposition reaction was carried out.

 Tenofovir ester (DEPMPA) obtained in the above procedure was dissolved in 200 ml of acetonitrile, and 120 ml of bromotrimethylsilane was added thereto to effect a reflux reaction. After 5 hours reaction, the reaction was completed (14 hours) while checking the reaction process by TLC and HPLC. The reaction solution was diluted with 200 ml of cold purified water and then washed twice with 200 ml of ethyl acetate to remove the trimethylsilane component. The aqueous solution was concentrated under reduced pressure at room temperature to form a residual solvent amount in the vessel, Respectively.

Next, the aqueous solution was neutralized with 50% sodium hydroxide to adjust pH to 3. The resultant crystals were filtered and washed with cold purified water (5 캜) to obtain a purified water (H ₂ O). The crystals were dried at 65 ° C under reduced pressure (30 Torr) to obtain 35.2 g of the crystals (yield: 55.6%). The crystals obtained above were again dried at 100 ° C under reduced pressure (30 Torr) to finally obtain 33 g of teporbir crystals (anhydrous yield: 55.4%). Tenofovir disoproxyl (Teno-D) and tenofovir disiproxyl fumarate (Teno-DF) were synthesized using the above anhydrous tenofovir in accordance with the conventional method described in the technical literature and the like.

 [Comparative Example 2]

Synthesis of Tenofovir

(Use of magnesium tertiary butoxide catalyst) (Ref: Published US2009 / 0286981 A1)

40 g (0.21 mol) of hydroxypropyl adenine is dissolved in 80 ml of dimethylformamide (DMF), 28.4 g (0.167 mol, 0.79 Equiv.) Of magnesium t-butoxide is added and suspended. 80 g (0.2486 mol, 1.18 Equiv.) Of diethyl-p-toluenesulfonyloxymethylphosphonate (DEPTSMP) was added dropwise over 21 hours while the reaction temperature was maintained at 74 DEG C, and further reaction was carried out at 80 DEG C for 5 hours And the reaction progress was confirmed by TLC and HPLC (takes 26 hours).

After the reaction was completed, the mixture was cooled to 20 ° C, and 24 g (0.4 mol) of glacial acetic acid was added thereto to adjust the pH to 6-6. Then, the reaction solution was completely distilled off under reduced pressure at 80 占 폚. The residue reaction solution was cooled to 20 ° C, 240 ml of methylene chloride (MC) and 40 ml of purified water were added, and the resulting inorganic substance was filtered, and the organic layer was separated. The aqueous layer and the inorganic residue were further extracted with 100 ml of MC and filtered at the same temperature to remove the inorganic substance. The organic solvent was added thereto, followed by washing with 50 ml of cold purified water at 10 ° C or less, followed by washing with 50 ml of a saturated sodium chloride solution, After dehydration, the organic layer was distilled under reduced pressure to give terpovir ester (DEPMPA). To the residue was added 200 ml of toluene, distilled under reduced pressure to reconstitute the residual DMF, and the remaining DEPMPA was hydrolyzed by the dealkylation reaction.

260 ml of HBr aqueous solution was added to the prepared teppovir ester (DEPMPA) and stirred at 90 ° C for 5 hours. After 5 hours of reaction, the reaction was completed by TLC and HPLC, respectively (10 hours ). 120 ml of cold purified water was carefully added to the reaction solution, and the solution was diluted with 120 ml of MC to remove the trimethylsilane component. The aqueous solution was concentrated under reduced pressure at room temperature to obtain a residual amount of MC in the vessel, To be 5 [deg.] C.

Next, the cooled aqueous solution was neutralized with 40% sodium hydroxide solution to adjust the pH to 2.5 to 3.0. Crystals produced during the above process were filtered and washed with cold purified water (5 ° C) to obtain tenofovir crystals. The crystals were dried under reduced pressure (30 Torr) at 65 deg. C to obtain 44 g (yield: 70%). The crystals were further dried under reduced pressure at 100 ° C. (30 Torr) to obtain 41 g of terpovorin crystals (anhydrous yield: 69%). The above anhydrous terpovorin was used to prepare tenopovir disoproxil Tenofovir disoproxyl fumarate (Teno-DF) was synthesized according to the conventional method described in the technical literature and the like.

 [Comparative Example 3]

Synthesis of Tenofovir

(Use of sodium amide and magnesium chloride complex catalyst) (Ref: published patent application US 2013/0165413 A1)

40 g (0.21 mol) of hydroxypropyl adenine is dissolved in 80 ml of dimethylformamide (DMF). The reaction temperature was maintained at 0 ° C and 16.16 g of sodium amide was added. After stirring for 30 minutes, 19.7 g of magnesium chloride was added at 25 ° C, and the mixture was stirred for 1 hour. To the mixture was added 120 ml of toluene, 100 g (0.31 mol, 1.48 Equiv.) Of sulfonyloxymethylphosphonate (DEPTSMP) was added dropwise over 4 hours. The reaction was further maintained at 80 ° C for 5 hours while stirring, and the progress of the reaction was confirmed by TLC and HPLC (Takes about 16 hours).

 After completion of the reaction, the reaction solution was completely distilled off under reduced pressure at 70 占 폚. The residue reaction solution was cooled to 20 ° C, 240 ml of methylene chloride (MC) and 40 ml of purified water were added, and the resulting inorganic substance was filtered, and the organic layer was separated. The aqueous layer and the inorganic residue were further extracted with 100 ml of MC and filtered at the same temperature to remove the inorganic substance. The organic solvent was added, and the organic layer was washed with 50 ml of saturated sodium chloride solution The organic layer was dried under reduced pressure to give terpovir ester (DEPMPA). 100 ml of toluene was added to the residue, and the remaining DMF was distilled off under reduced pressure to carry out a hydrolysis reaction in which DEPMPA was dealkylated.

 260 ml of HBr aqueous solution was added to the above obtained terpovir ester (DEPMPA) and stirred at 90 ° C for 5 hours. After 5 hours of reaction, the reaction was completed by TLC and HPLC. To the reaction mixture, 120 ml of cold purified water was carefully added, diluted, and then washed twice with 120 ml of MC to remove the trimethylsilane component. The aqueous solution was concentrated under reduced pressure at room temperature to obtain a residual amount of MC in the container. Respectively.

Next, the pH was adjusted to 2.5 to 3.0 by neutralizing with 50% sodium hydroxide. The resulting crystals were filtered, washed with cold purified water (5 ° C) and again washed with 40 ml of cold acetone to obtain tenofovir 1 crystal water (H ₂ O). The crystal is heated at 90 DEG C for 30 minutes with 360 mL of purified water, cooled to 0 DEG C and filtered after 4 hours. The crystals were dried at a dryer temperature of 65 DEG C under reduced pressure (30 Torr) to obtain 36 g of a crystal (yield: 57%). The crystals were dried at 100 ° C under reduced pressure (30 Torr) to finally obtain 33.7 g of a terpinovir anhydride crystal (anhydrous yield: 56.6%).

 With the anthophyll anhydride, the following reaction was carried out with tenofovir disoproxil

(Teno-D) and Tenofovir disiproxyl fumarate (Teno-DF) were synthesized according to the conventional methods described in the literature.

 [Example]

Synthesis method of Tenofovir (Synthesis method according to the present invention)

(Magnesium t-butoxide, calcium hydride complex catalyst and method of using ion exchange resin)

40 g (0.21 mol) of hydroxypropyladenine (R) -9- (2-hydroxypropyl) adenine and HPA were dissolved in 200 ml of dimethylformamide (DMF), 143.2 g (0.84 mol) of magnesium t-butoxide, 4,0 Equiv.) And 4.42 g (0.105 mol) of calcium hydride as a reaction catalyst. 100 g (0.325 mol, 1.5 Equiv.) Of diethyl-p-toluenesulfonyloxymethylphosphonate (DEPTSMP) was added over 2 hours at a reaction temperature of 30 to 35 DEG C and the mixture was reacted at 80 DEG C for 5 hours or more The reaction progress is confirmed by TLC and HPLC (takes 7 hours).

 After completion of the reaction, glacial acetic acid is added dropwise at 20 ° C to adjust pH to 6-6, which is slightly acidic. Ethyl acetate (1,000 ml) is added to the reaction mixture, the mixture is heated to 50 to 60 占 폚, cooled to 40 占 폚 and filtered. 500 ml of ethyl acetate is further added to extract the inorganic residue and the filtrate is removed at the same temperature to remove the inorganic matter. The organic solvent is added thereto and the solvent is distilled off at 70 ° C or lower. The organic layer was washed with an additional 100 ml of cold water twice, followed by washing with 50 ml of a saturated sodium chloride solution (sodium chloride solution), and then the organic layer was dehydrated with anhydrous sodium sulfate. The organic layer was dried over anhydrous magnesium sulfate, The organic layer is vacuum-dried to obtain terpovir ester (DEPMPA). 300 ml of toluene was added to the residue, and the remaining DMF and ethyl acetate were distilled off under reduced pressure. The residue was distilled under reduced pressure with 300 ml of cyclohexane to reconstitute ethyl acetate and DMF. The DEPMPA residue was subjected to hydrolysis Respectively.

 The above obtained terpovir ester (DEPMPA) was dissolved in 200 ml of acetonitrile, and 120 ml of bromotrimethylsilane was added to perform reflux reaction. After 5 hours of reaction, the reaction was completed by examining the reaction process by TLC and HPLC (12 cycles). The reaction solution was carefully diluted with 200 ml of cold purified water and then washed twice with 200 ml of ethyl acetate to remove the trimethylsilane component. The aqueous solution was concentrated under reduced pressure at room temperature to obtain the remaining amount of the solvent in the container. Respectively.

The solution was neutralized with 40% sodium hydroxide to adjust the pH to 3.5. Conversely, the pH was precisely adjusted to 3.2 using an ion resin Dowex 50W cation exchange resin, and the resulting crystals were filtered. The crystals were washed with cold purified water (5 캜) to obtain tenofovir crystals. This crystal was dissolved in 400 ml of methanol and filtered to remove the ionic resin and concentrated under reduced pressure. The residue crystals were dried at a dryer temperature of 65 ° C under reduced pressure (30 Torr) to obtain 48.0 g of terpovorin crystals (yield: 75.9%). The crystals were dried under reduced pressure at 95 ° C (30 Torr) to finally obtain 45 g of anhydrous tehnoporbir crystals (anhydrous yield: 75.7%).

 Tenofovir disoproxil (Teno-D) was synthesized by the following reaction with the above anhydrous terfenovir.

[Method for synthesizing Tenofovir disuproxyl (Teno-D)] [

 25 g of the terpovinyl anhydride obtained in the above example was added to 100 ml of N-methyl-2-pyrrolidinone (NMP) and 50 ml of toluene, distilled under reduced pressure at 60 ° C to remove moisture, and again 50 ml of toluene was distilled off.

The reaction solution was cooled to 35 DEG C and 35.2 g (4 equiv.) Of triethylamine was added to the reaction mixture to form crystals. The mixture was homogenized and suspended with stirring, 28 g (1.0 Equiv.) Of tetrabutylammonium bromide was added And heated to 45 캜.

To this was added 66.4 g (5.0 Equiv.) Of chloromethylisopropyl carbonate and stirring was continued at 45 to 55 占 폚. After 5 hours, the progress of the reaction was monitored by TLC and HPLC. When the reaction was completed and the viscous liquid turned transparent red, it was cooled to room temperature, and 150 ml of cyclohexane was added thereto. After stirring vigorously, Removed and again washed with 100 ml of cyclohexane to remove. 300 ml of ethyl acetate and 100 ml of cold water were added to the reaction layer and stirred to remove the aqueous layer. The aqueous layer was washed twice with 50 ml of ethyl acetate and added to the undiluted solution. The organic layer was washed with 50 ml of cold water and 50 ml of saturated brine, dehydrated with magnesium sulfate, and then vacuum-dried to obtain 53 g of crystalline sludge, Tenofovir disoproxil. The product was further cooled at a lower temperature for crystallization, and the content was analyzed by HPLC (purity: 85%, actual amount: 45 g, area: 96%).

 [Synthesis of Tenofovir disisoproxyl fumarate (Teno-DF)

 (0.087 mol) was dissolved in 120 ml of isopropyl alcohol, and 12.5 g (0.107 mol, 1.2 Equiv.) Of fumaric acid was added thereto. The mixture was stirred at 50 to 55 ° C for 2 hours Respectively. The reaction solution was cooled to 3 to 5 DEG C, and the resulting crystals were sufficiently stirred and then filtered. The crystals were suspended in 250 ml of ethyl acetate and stirred for 1 hour. The reaction solution was warmed to 10 DEG C or lower, filtered and washed. The crystals obtained in this way were vacuum-dried at 40 DEG C to obtain 35 g of terpovorbidisoproxyl fumarate (HPLC area Purity 98.5%, yield 63.6%).

 [Production method of film-like preparation]

Furtherance Example 1 Example 2 Example 3
Tenofovir Distemoloxil
Fumarate 37.5%
(Tenofovir disiproxyl
Fumarate as 150 mg) 37.5%
(Tenofovir disiproxyl
Fumarate as 150 mg) 37.5%
(Tenofovir disiproxyl
Fumarate as 150 mg) Hydroxypropyl
Methylcellulose
-
20.00%
20.00% Pullulan 35.0% 15.00% 15.00% Polyethylene glycol 600 15% 15% - Polyethylene glycol 400 - - 15% glycerin 2% 2% 2% Polysorbate 20 0.10% 0.10% 0.10% Acesulfame 10.40% 10.40% 10.40% Purified water dose dose dose sub Total 100% 100% 100% Gross weight (mg / h) 400 mg 400 mg 400 mg

(The content of each component in the above table means the weight% with respect to the total weight)

Example 1

 1. Add polyethylene glycol 600, glycerin, polysorbate 20 and acesulfame to purified water and dissolve completely.

 2. Pullulan is added to the solution of step 1 and completely dissolved to prepare a mixed solution.

 3. The mixture is aged at room temperature (25 ° C) for 12 hours or more.

 4. Add the main component, terpoviridisoproxyl fumarate, to the mixture and stir well.

5. Apply the above mixed solution so that the main component is contained in an amount of 150 mg in an area of 12 cm ² .

 6. After applying, dry at 65 ° C for 30 minutes and at 45 ° C for 3 hours or more.

Example 2

 Except that the pullulan content was changed to 15% by weight and the hydroxypropylmethylcellulose content was changed to 20% by weight.

Example 3

 Except that polyethylene glycol 400 was used instead of polyethylene glycol 600 and the content thereof was changed to 15% by weight.

 [Sexuality Test and Analysis Method and Evaluation of Film]

1) Sensory test

 1: little bitter, 2: little bitter, 3: slightly bitter, 4: bitter, 5: very bitter

2) Bending test

 The bending test was carried out using two fingers after the film was folded in half and repeatedly folded until it was cut in half. After 30 minutes of opening the film, the temperature and humidity were measured at 22 ° C and 55 ° C % RH. The higher the number, the better the brilliance.

3) Dissolution test

 General Test Methods for Korean Pharmacopoeia The test is carried out using an assisting plate in accordance with the General Release Method. When tested according to the above test method, the dissolution time should not be more than 3 minutes.

4) Peel test

 The peel test measures the degree of peeling from the PET film as the support film.

 1: very poor peeling, 2: poor peeling, 3: peeling is normal, 4: peeling is good, 5: peeling is very good

5) Film Evaluation Table

Example 1 Example 2 Example 3 Sensory test 3 3 3 bending test 3 5 8 Dissolution test Less than 3 minutes Less than 3 minutes Less than 3 minutes Peeling test 5 5 5

Test and analysis methods and evaluation

<Synthesis HPLC data: Comparison of Tenofovir and Tenofovir-DF Compositions HPLC Data>

 Tenofovir Compound

Tenofovir standard

Teno-DF compound

Teno-DF standard product

[Film Formulation HPLC Analysis Evaluation Data: 24 week stability HPLC data]

The content test of Example 1 (Tenofovir DF standard)

The content test of Example 1 (Tenofovir DF film)

The 4-week stability test (Tenofovir DF standard product)

The four-week stability test (made by Tenofovir DF film) of Example 1

The 12-week stability test (Tenofovir DF standard)

The 12-week stability test of Example 1 (Tenofovir DF film)

The 24-week stability test (Tenofovir DF standard)

The 24-week stability test (made by Tenofovir DF Film)

[Improvement and comparative evaluation of synthetic method of tenofovir and comprehensive evaluation of film formulations]

In this synthesis method (Example), 45 g (the weight of the final product, tenofovir crystal) of 40 g (the weight of the HPA as a starting material) is produced based on the amount of raw material used when synthesizing tenofovir in HPA, 112.5%) was obtained, which was superior to the 0.825 ratio (82.5%, 33/40 = 0.825) of Comparative Example 1 and superior to the 1.03 ratio (103%, 41.2 / 40 = 1.03) of Comparative Example 2 (84.2%, 33.7 / 40 = 0.842) of Comparative Example 3, which was increased by 9.5%, 69% -> 75.7% = 6.7%

 In addition, Tenofovir disoproxil fumarate (Teno-DF) synthesized by the synthetic method according to the present invention, which has been synthesized through intermediate steps of Tenofovir disoproxyl (Teno-D) with Tenofovir (anhydrous) ), The yield of the synthetic step of the present invention is 1.4% (140%), which is 35 g at 25 g based on the amount of the raw material of the present invention, (138%, 69/50 = 1.36) of the combined yield ratio shown in Development (2010, Vol.14, 1194-1201).

 Therefore, it has been found that the synthesis method according to the present invention, which can produce tenofovir in high yield, is superior and effective than the conventional synthesis method.

 It has also been found that the film formulation of medicines using the above-synthesized terfenoviridisoproxyl fumarate is also stable. As a result, it is possible to develop medicines using terpovoridylsofosylxyl fumarate synthesized according to the present invention as a novel film-type product .

Claims (5)

A method for producing a therapeutic agent for viral infection, comprising administering to a patient an effective amount of at least one stabilizing agent selected from the group consisting of hydroxypropylmethylcellulose and pullulan, And a method for producing a therapeutic agent for viral infection using the present invention as an effective ingredient The method according to claim 1, wherein 0.1 to 10 parts by weight of hydroxypropyl methylcellulose is added to the total weight of the infectious agent for therapeutic treatment of viral infection comprising an effective amount of GS- The pharmaceutical composition according to claim 1 or 2, which comprises 5 to 35 parts by weight of pullulan based on the total weight of the infectious agent, and 1 to 15 parts by weight of polyethylene glycol, wherein the effective amount of terpoviridisoproxyl fumarate For the preparation of a therapeutic agent for viral infection The method according to claim 3, wherein the polyethylene glycol is any one selected from polyethylene glycol 400 and polyethylene glycol 600. Description: TECHNICAL FIELD [0001] The present invention relates to a method for producing a therapeutic agent for viral infection comprising an effective component of GS- 5. The oral cavity film formulation according to claim 4, wherein the orally dissolvable film formulation has a thickness of 10 to 400 micrometers, and the content of tenofovir disoproxyl fumarate is 7.5 to 30 mg / A method for producing a therapeutic agent for viral infection using roxil fumarate as an active ingredient