KR101439255B1 - The New Tenofovir disoproxil salt and the preparation method thereof - Google Patents

Abstract

The present invention relates to novel tenofovir disoproxil aspartate compounds.
The salt compound can minimize the generation of a flexible substance according to changes over time as compared with the terbinafide disoproxil fumarate salt, so that the stability of the preparation can be improved by greatly reducing the amount of impurities generated during storage of the product containing the salt compound , Eliminating the need to perform separate studies of toxic effects. In addition, since the salt compound of the present invention greatly improves the stability, the wetting property and the solubility according to the pH change and has a physico-chemical property, it can be used for the treatment of HIV-1 infection and chronic hepatitis B together with a pharmaceutically acceptable carrier May be used as an active ingredient of a pharmaceutical composition useful in the present invention.

Description

[0001] The present invention relates to a novel salt of tenofovir disoproxil salt and a preparation method thereof,

The present invention relates to a novel salt of tenofovir disoproxyl and its preparation.

Tenofovir dysoproxyl fumarate (hereinafter referred to as TDF) has the chemical name 9- [2- ( R ) - [[bis [[(isopropoxycarbonyl) oxy] methoxy] phosphinoyl] methoxy] Adenine fumaric acid (1: 1), a prodrug that is hydrolyzed upon absorption, and is a phosphonomethoxynucleotide analog useful in the treatment of HIV-1 infection and chronic hepatitis B.

TDF was approved and approved by the United States Food and Drug Administration as a hepatitis B treatment in Europe in April 2008 and in November 2008. It was approved by the United States Food and Drug Administration received. Tenofovir, an active metabolite of TDF, has similar structure and efficacy to adefovir (Hexera) but has a stronger B-response in lamivudine-resistant patients (300 mg) than adefovir (10 mg) Hepatitis virus DNA. In addition, Tenofovir is an antiviral drug belonging to the FDA's FDA's Classification of Fetal Risk Category B (drug without risk of fetal risk in animal studies).

Despite these advantages, it is difficult to secure the physico-chemical stability of Tenofovir, and the original developer has conducted various studies to overcome it. For example, it is known that tenofovir disoproxil (hereinafter referred to as TD) has a serious problem in stability due to the influence of moisture. Published Research, 2001, 18, 234-237; Pharmaceutical Research, 2000, 17, 1098-1103 ) in the TD is hydrolyzed in the presence of water to generate formaldehyde, the generated formaldehyde and TD N ⁶ - amine groups cause a condensation reaction impurities, tenofovir D. Thorpe It has been reported to produce a lock polymer (Tenofovir disoproxil Dimer).

Generally, in order to exhibit a certain effect at all times, it is necessary not only to suppress the decrease of the content of the active ingredient in the storage period as well as immediately after the production of the product, but also to minimize the increase of the decomposition product of the active ingredient, . Therefore, prevention of impurities in medicines is very important for the quality control of medicines.

As for the purity test, the Korean Pharmacopoeia sets the purity test item separately, and the US Pharmacopoeia is defined as 'Ordinary impurities'. Unless otherwise specified, Is 2.0% or less, or regulates the amount of each of the flexible substances in related compounds and chromatographic purity in each medicine. In European Pharmacopoeia and UK Pharmacopoeia, it is defined as related substances. In Japanese Pharmacopoeia, it is regulated as a flexible substance in the purity test and regulates its amount.

In addition, ICH Guideline Q3A, which is an international agreement on medicines, describes general information on the preparation of new medicines and Q3B provides general information on the specification of impurities in new medicinal products. Among these, the impurity specifications in the product are further subdivided into Reporting Thresholds, Identification Thresholds, and Safety Validation Thresholds. The report threshold is simply a brief report of the detected substance itself. The structure identification threshold should identify the physicochemical structure, and the safety validity threshold should be determined by evaluating not only the physicochemical structure but also the toxicological effects, And to provide a basis for judging whether or not the human body is safe due to the impurities. In other words, it can be said that there is a great difference in the width of the data that must be provided according to the increase of the trace amount of impurities, and that the possibility of the toxic hazard can be greatly increased.

Thus, the management of impurities contained in medicines is treated as a very important issue so as to establish common agreements not only for individual countries but also internationally.

Therefore, the ability to maintain the incidence rate of impurities during the storage period of a drug containing a tenofovir disoproxil can be prevented from further physicochemical structural identification and / or toxicological studies on the corresponding substance, However, it can be said that it is significant in that it can largely eliminate the toxicological danger that may be caused by the impurities. Therefore, the medicament containing tenofovir disoproxil should be able to maintain the impurity generation rate during the storage period to a certain level or less.

Gillard, the original developer of Tenofovir, is suitable for oral administration, and is capable of inhibiting the production and increase of polymer impurities by inactivating the N ⁶ -amine group, (U.S. Pat. No. 5,935,946), International Patent Application No. 2010 / 142,761 discloses a method for improving the processability as a tablet formulation by reducing the wettability and improving the solubility and securing stability of the tablet of the present invention. Disclose terpovoid disoproxil succinate.

The inventors of the present invention have made efforts to investigate a novel salt capable of minimizing the generation of a flexible substance while having physico-chemical advantages equal to or higher than that of the existing tenofovir disoproxil fumarate salt. As a result, Sooproxyl aspartate was developed.

It is an object of the present invention to provide a novel tenofovir disoproxil salt compound which has physico-chemical advantages equal to or higher than that of terfenoviride soaproxyl fumarate and can minimize the generation of a flexible substance.

According to the above object, the present invention provides a pharmaceutical composition for treating HIV-1 infection and chronic hepatitis B, which is pharmaceutically acceptable and has improved physico-chemical conditions, ( R ) - [[bis [(isopropoxycarbonyl) oxy] methoxy] phosphinoyl] methoxy] propyl] adenine aspartic acid (hereinafter referred to as terfenoviridisoproxylaspartate) to provide.

≪ Formula 1 >

The tenofovir disoproxil aspartate of the present invention is present as a salt material produced by ion-binding 1: 1 of aspartic acid and terfenoviride disoproxil free base.

The tenofovir disoproxil aspartate according to the present invention can be prepared by reacting aspartic acid with tenofovir disoproxil free base to produce tenofovir disoproxil aspartate. This can be illustrated by the following Reaction Scheme 1.

<Reaction Scheme 1>

Detailed Description of the Invention The process for the preparation of tenofovir disoproxil aspartate according to the present invention will be described in detail.

1) dissolving terbinafide disoproxil free base in water;

2) adding aspartic acid to a solution in which the tepofovir disoproxil free base is dissolved in the first step, followed by heating and stirring; And

3) Tenofovir disoproxylaspartate can be prepared from the step of crystallizing the solution heated and stirred in the second step by cooling and stirring at room temperature or below room temperature to obtain terfenoviride soaproxyl aspartate have.

In the present invention, the Tenofovir disoproxil free base may be prepared directly or by a known method, and water is used as a reaction solvent.

The aspartic acid used in the production method of the present invention is preferably used in an amount of 1 to 3 equivalents, more preferably 1.1 to 1.5 equivalents, based on the terpovoid disoproxil free base. In addition, in the present invention, water used as a reaction solvent is preferably used in an amount of 5 to 100 ml per 1 g of GS-free isodisulfroxylic free base, more preferably 10 to 50 ml per 1 g of GS-free GS Do.

In the production process of the present invention, at the time of heating and stirring The reaction temperature is from room temperature to the boiling point of the reaction solvent at a temperature of 100 DEG C or less, preferably 40 to 60 DEG C. [

In the production process of the present invention, the reaction time during heating and stirring may vary depending on the reaction temperature, but is preferably from 10 hours to 15 hours.

In the present invention, the crystallization method means a known crystallization method. For example, the mixture is cooled to a room temperature (20 to 30 ° C) or a room temperature or lower, and then the mixture is stirred or allowed to crystallize.

The tenofovir disoproxil aspartate according to the present invention can minimize the generation of the suppository due to the change over time as compared with the terpovoid disoproxil fumarate, so that the amount of impurities generated during the storage of the product containing the salt compound Significantly lowering the stability of the formulation and eliminating the need to perform separate studies of toxicological effects. In addition, since the salt compound of the present invention greatly improves the stability, the wetting property and the solubility according to the pH change and has a physico-chemical property, it can be used for the treatment of HIV-1 infection and chronic hepatitis B together with a pharmaceutically acceptable carrier May be used as an active ingredient of a pharmaceutical composition useful in the present invention.

In addition, even when the preparation of terfenoviride soaproxyl aspartate is carried out by a simple manufacturing process, it is possible to obtain a more pure terfenoviride sofferoxil in the course of synthesizing the acid addition salt.

FIG. 1 is a graph showing the pH-dependent stability results of the control group, tenofovir disoproxil fumarate,
FIG. 2 is a graph showing the stability results of the pH of the tablet of the present invention, pH-dependent,
Fig. 3 is an X-ray diffraction chart of the terpovoid disoproxyl fumaric acid salt,
4 is an X-ray diffraction chart of the tenofovir disoproxil aspartate,
5 is a differential scanning calorimetry spectrum of the terpovoid disoproxyl fumarate salt,
FIG. 6 is a differential scanning calorimetry spectrum of teporbidisoproxylaspartate,
7 is a ¹ H nuclear magnetic resonance spectrum of terpovoid disoproxil fumarate,
Fig. 8 is a ¹ H nuclear magnetic resonance spectrum of teporbidisoproxylaspartate. Fig.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Bruker UltraShield (TM) 400 (400 MHz) was used for ¹ H NMR in the following examples. Agilent 1200 series was used for HPLC, and Electrothermal IA9000 and Mettler Toledo DSC 823 were used for melting point. Reagents and solvents not specifically mentioned by the manufacturer were purchased from Aldrich, Sigma and Acros.

< Reference example  1> Tenofovir Dissociol Free base  Produce

In a 1,000 mL reaction portion, 50.0 g of tepofovir disoproxyl fumaric acid salt (purity: 98.60%, Luna chemicals; china), 250 mL of ethyl acetate and 250 mL of purified water were added and stirred and stirred at room temperature for 10 minutes. A saturated aqueous solution of sodium bicarbonate is added dropwise to the reaction part to adjust the pH to 7.0, and the mixture is stirred for 30 minutes and left for 10 minutes. Discard the water layer. The organic layer is dried over anhydrous magnesium sulfate, filtered and concentrated. When the volume of the concentrate reaches 125 mL, the solid starts to precipitate, and the concentration is stopped, followed by stirring at room temperature (2 hours). The temperature of the reaction part is cooled to 5 캜 and stirred for 2 hours. After filtration, the solution was dried at 45 캜 for 2 hours to obtain 35.0 g of a white colored title compound, terfenoviride soaproxyl free base.

HPLC purity: 99.61%

_{^{1 H NMR (400㎒, D 2}} O): 8.16 (s, 1H), 8.05 (s, 1H), 5.52-5.39 (m, 3H), 5.33-5.28 (m, 1H), 4.89-4.77 (m, 2H), 4.34-4.30 (m, 1H), 4.16-4.05 (m, 2H), 3.93-3.90 (m, 1H), 3.80-3.74

Melting point: 101.5 DEG C

&Lt; Example 1 > Preparation of tenofovir disoproxil aspartate

In a 2,000 mL reaction vessel, 50.0 g of terfenoviride soaproxyl free base, 750 mL of purified water and 15.4 g of L-aspartic acid (Aldrich) were added, and the temperature was raised to 46-51 캜 and stirred for 2 hours. After cooling to room temperature, the mixture was stirred for 15 hours. After filtration, the filtrate was washed with 50 mL of purified water and dried at 45 ° C for 2 hours to obtain 50.6 g of a white colored title compound, terfenoviride soaproxyl aspartate.

HPLC purity: 99.68%

_{^{1 H NMR (400㎒, D 2}} O): 8.30 (s, 1H), 8.18 (s, 1H), 5.54-5.42 (m, 3H), 5.39-5.34 (m, 1H), 4.91-4.83 (m, 2H), 4.42-4.38 (m, IH), 4.23-4.18 (m, IH), 4.12-4.06 2.86 (m, 1H), 2.82-2.75 (m, 1H), 1.28-1.24 (m, 15H) ppm

Melting point: 109 DEG C

< Experimental Example  1> Flexible material  Evaluation of stability according to the degree of occurrence (acceleration, harshness)

Stability test of pharmaceuticals refers to a test to evaluate the stability of quality due to changes over time in order to set the storage method and the period of use of medicines, etc. After establishing appropriate standards, the pharmaceuticals are analyzed on the basis of the prescribed test method, and the significant changes are evaluated, and the validity period is determined by judging whether or not the standard conforms. Therefore, ensuring the appropriate stability of the drug is one of the most important factors in the commercialization of the drug.

Therefore, in order to evaluate the stability (acceleration, harshness) according to the degree of occurrence of the soft matter of the cannabisidisoproxylaspartate of the present invention (Example 1), tenofovir disoproxil fumarate (Luna chemicals; china) Were subjected to acceleration and harshness stability tests according to the ICH guidelines and analyzed using a high performance liquid chromatography (HPLC) analysis method described in USP (USP). The results are shown in Tables 1 and 2 Respectively.

As can be seen from Tables 1 and 2, it can be seen that the tenofovir disoproxylaspartate of the present invention is a very stable substance in comparison with terbinafide desoperoxyl fumarate under accelerated and harsh conditions. In particular, since the flexible substance generated in the tenofovir disoproxil fumarate is hardly generated in the tenofovir disoproxil aspartate of the present invention, high purity can be maintained, There is no problem due to decomposition products that may occur. Furthermore, in the case of Tenofovir isoproxil monoester, the major degradation product, tenofovir disoproxil fumarate increased to 1.54% (30 days) at 60 ° C harsh conditions, while tenofovir disoproxil aspartate increased to 0.271% , And the stability was greatly improved by physicochemical properties.

Therefore, it can be seen that the tenofovir disoproxil aspartate according to the present invention is a drug which can keep a high purity for a long time by minimizing the generation of a flexible substance and is easy to store and has excellent storage stability.

< Experimental Example  2> pH  Evaluation of stability according to change

When the drug is orally administered, the drug is absorbed into the body through the stomach, duodenum, small intestine, and the like. Therefore, ensuring adequate stability according to the pH change until the drug is absorbed in the body has a great influence on the elution amount of the drug, which is an important factor that determines the drug efficacy in the end.

Therefore, a pH buffer solution (pH 1.2, pH 3.2, pH 5.2, pH 7.2, pH 9.2) was prepared to evaluate the stability of each drug according to the behavior of the drug in the body. Then, tenofovir disoproxil aspartate of the present invention was used as a control and tenofovir disoproxil fumarate was used as a high concentration (5 mg / mL), and the solution was completely dissolved in a mixture of acetonitrile and water (1/19) The pH of the buffer solution prepared above was diluted with buffer solution to maximize the exposure to the pH buffer solution. The samples were placed in a 40 ° C chamber, and the time-dependent changes in the contents were measured. The reaction was stopped with a mixture of acetonitrile and water (1/19) in water, Filtered through a 0.2 um filter and quantitated using high performance liquid chromatography (HPLC).

The pH-dependent stability results of tenofovir disoproxil aspartate and tenofovir disoproxil fumarate are shown in FIGS. 1 and 2, respectively.

As can be seen from FIGS. 1 and 2, the tenofovir disoproxil aspartate according to the present invention has stability equivalent to that of the existing commercialized terfenoviridisofroxyl fumarate salt at various pHs. In the basic environment, both of the substances are rapidly degraded due to the nature of the drug (hydrolyzed prodrug).

< Experimental Example  3> An assessment of conventionality

Since the low specificity is a very important factor for the processing and storage of actual medicines among the physical properties required for the compound to be used as a raw material for medicines, the degree of customary properties of the present invention is evaluated with regard to the tenofovir disoproxil aspartate.

Tenofovir disoproxil aspartate (500 mg) was placed in a glass tube for the measurement of humor and dried at 25 ° C for 12 hours under nitrogen. After 12 hours, the mass change rate of the sample was automatically measured at a relative humidity of 15, 35, 55, 75 and 95% using a conventional measuring device (Model: Hydrosorb 1000, manufacturer: Quantachrome Instruments) Tenofovir disoproxil fumarate was used as a control.

According to the Korea Food and Drug Administration (KFDA) standards, tenofovir disoproxil fumarate is controlled and regulated to have a water content of 1.0% or less. As can be seen from Table 3, the control group, Tenofoviride soaproxyl fumaric acid salt, was relatively poor in the properties of the moisture due to the rapid moisture content of the air as the relative humidity was increased.

However, it can be seen that the tenofovir disoproxylaspartate according to the present invention exhibits a very low specific humidity of about 0.2% at the entire range of the experiment conducted, and can be stably stored even if the raw material is exposed to moisture in the air have.

Therefore, the inventive tenofovir disoproxil aspartate exhibits excellent non-humectant properties over tenofovir disoproxil fumarate, which is excellent in storage stability and can be stored for a long time and can be further easily commercialized.

< Experimental Example  4> Solubility assessment

Solubility is a very important property of a drug. Even if the drug is very excellent, product development often fails due to extremely low solubility. That is, when the solubility is low, it precipitates and is present in a precipitated state, which is a great factor for reducing oral absorption. Therefore, ensuring adequate solubility is an essential factor in the commercialization of drugs.

The degree of solubility of the present invention to the tenofovir disoproxylaspartate was evaluated and is shown in Table 4 below. Tenofovir disoproxil fumarate was used as a control.

50 mg of teporbidisoproxylaspartate was added to water, pH 1.2, pH 4.0, pH 6.8 solution, and shaken at 25 ° C and 150 rpm for 3 days. After 3 days, each sample was filtered with a 0.45 mu m filter, diluted with a mixture of acetonitrile and water (1/19), analyzed by high performance liquid chromatography (HPLC) and quantified.

As can be seen from the results of Table 4, the inventive tablet of Tropofiber disoproxylaspartate showed equivalent or higher solubility than that of the existing commercialized terfenoviride soaproxyl fumaric acid salt, In 4.0 (duodenum), the solubility is about twice that of tenofovir disoproxil fumarate, which greatly improves the physicochemical properties of the drug, which is very advantageous for the absorption and dissolution of the drug.

< Experimental Example  5> Crystalline polymorphism ( XRD )

The X-ray powder diffraction pattern is a unique characteristic of a drug and is widely used for distinguishing a crystal form and a hydrate. Therefore, the X-ray powder diffraction pattern of the present inventive Tenofovir disoproxil aspartate and the existing commercialized Tenofovier disoproxyl fumarate was measured.

The X-ray powder diffraction pattern of the two materials was measured at a scan rate of 0.020 ° 2-theta (2θ) per minute using a Bruker AXS GmbH model D8 Advance X-ray diffractometer and an evaluation software system interface. And scanned between 3 and 40 ° 2θ by exposure to an X-ray generator operating at 40 kV and 40 mA using a standard line focus copper x-ray tube (Siemens) equipped with a one-dimensional semiconductor X-ray detector using a silicon strip. The weighted average value of the X-ray wavelength used in the calculation was Cu-Ka1.541838.

As a result of the measurement, the tenofovir disoproxyl fumaric acid salt was observed at 4.9 °, 10.2 °, 10.8 °, 18.2 °, 21.9 °, 24.0 °, 25.0 °, and 25.4 ° almost in the same manner as described in the conventional patent (US Pat. No. 5,935,946) , 27.9 °, 30.1 ° and 30.5 °, respectively. In contrast, tenofovir disoproxil aspartate exhibited a characteristic curve at 7.4 °, 13.0 °, 14.5 °, 18.2 °, 19.2 °, 19.4 °, 19.9 °, 20.9 °, 22.9 °, 26.7 °, 27.1 ° and 28.1 ° Ray powder diffraction peaks showing that the two materials are different from each other.

< Experimental Example  6> Differential scanning calorie ( DSC )

Differential scanning calorimetry is also widely used as a means of identifying drug properties. Thus, thermal analysis of the tenofovir disoproxil aspartate and tenofovir disoproxyl fumarate of the present invention was carried out. The thermal analysis was obtained using a scan rate of 5 ° C per minute under a nitrogen atmosphere. Calorimetric scans were obtained using a differential injection calorimeter (Mettler Toledo, DSC 823e) and the amount of sample used was 3.0 mg. As expected, it was confirmed as shown in Table 5 that the starting and characteristic peaks of the two materials were different from each other.

Claims (6)

Which is 9- [2 [( R ) - [[bis [(isopropoxycarbonyl) oxy] methoxy] phosphinoyl] methoxy] propyl] adenine aspartic acid.
&Lt; Formula 1 >

delete The method of claim 1, wherein the compound of formula (1) is at least one selected from the group consisting of 7.4 °, 13.0 °, 14.5 °, 18.2 °, 19.2 °, 19.4 °, 19.9 °, 20.9 °, 22.9 ° Deg.], 26.7 [deg.], 27.1 [deg.] And 28.1 [deg.]. delete delete delete

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