KR101548724B1

KR101548724B1 - Antiviral Compounds in the Form of Solid and Method for Preparing thereof

Info

Publication number: KR101548724B1
Application number: KR1020140151114A
Authority: KR
Inventors: 지준홍; 유형철; 김재선; 엄기안; 이정범; 이보람; 김봉; 노광영
Original assignee: 주식회사 휴온스; 지준홍; 제이투에이치바이오텍 (주)
Priority date: 2014-04-25
Filing date: 2014-11-03
Publication date: 2015-09-02
Also published as: WO2015163724A1

Abstract

The present invention relates to a novel crystal form of a tenofovir disoproxil free base. According to the present invention, the tenofovir disoproxil free base ensures to enhance a stability of a medicine by reducing an impurity production during storage by being able to minimize a production of impure elements due to changes by a time flow compared to a tenofovir disoproxil fumarate or a previously known tenofovir disoproxil free base. In addition, the novel crystal form of the tenofovir disoproxil free base has an improved hygroscopicity, an excellent solubility in each pH, and physicochemically outstanding properties, thereby being able to be used as a useful active ingredient for a pharmaceutical composition.

Description

TECHNICAL FIELD [0001] The present invention relates to an antiviral agent and a method for preparing the same,

The present invention relates to a novel crystal form of tennofovirinoceptor free base and a process for its preparation.

Tenofovir Diisoproxil Fumarate is a compound having the chemical name 9- [2- (R) - [[bis [[(isopropoxycarbonyl) oxy] methoxy ] Phosphinoyl] methoxy] propyl] adenine fumaric acid (CAS Registry number 201238-50-9). The tennofovirino soaproxyl fumaric acid salt has a molecular weight of 635.52 and has the following structural formula 1).

Formula 1

Tenofovir isofloxyl fumarate is a nucleotide analog reverse transcriptase and HBV polymerase inhibitor developed by Gillian Adcience and marketed under the trade name Viread and is used in the treatment of HIV-1 infection and chronic hepatitis B It is a useful drug.

Tennophobia isoproxyl fumaric acid salt has an endothermic peak at about 118 ° as measured in differential scanning calorimetry (DSC) as a crystalline solid, as described in prior patent publications WO99 / 05150 and EP998480. Tennophobia isoproxyl fumarate is known to have several crystalline polymorphs, and crystal formations can occur due to formulation changes and aging. On the other hand, it is known that tennophobia soaproxil has a problem in stability due to the influence of water. Tenofoviridisoproxil is hydrolyzed in the presence of water to generate formaldehyde in Pharmaceutical Research, 18: 234-237 (2001); Pharmaceutical Research, 17: 1098-1103 (2000) , It has been reported that the formed formaldehyde and the amine group of tennophobia soaproxyl generate a condensation reaction to produce a tenofoviridisoproxil dimer, which is an impurity. Therefore, the preparation of tennofovirinosofroxyl fumarate is required to be strictly controlled in order to prevent moisture absorption, and to maintain dry conditions in packaging and storage.

According to the International Pharmacopoeia of the World Health Organization (WHO), the structure of several flexible substances in the case of tennofovirinoforosiloxyl fumarate is known, including optical isomers and hydrolysis products, hydrolyzed alkyl substituted substances, dimers Dimer), and partially hydrolyzed or all hydrolyzed of the dimer, and a combination of these materials. The storage conditions are as follows. Refrigerate in sealed and sealed containers at 2-8 degrees Celsius Condition. On the other hand, 9-prop-1-enyl-9H-purin-6-amine (9- (prop-1-enyl) It is regulated to be less than 5 ppm in the raw medicine.

In the prior published patent publications US2004 / 0018150, US6,465,649, US5,935,946, and US5,977,089, the synthesis of tennophobia isoproxyl and its salts is described. From the mucilaginous oil form tennofovirino sofferoxyl free base, Where the process is to prepare the soaproxyl fumarate salt or to produce the tropopino soaproxyl fumarate salt without isolating the tropopino soaproxyl salt.

Various problems may arise when the salt is formed by reacting tennophobia norsofloxil in the form of an oil and the salt formation reaction is carried out without isolation. Soproxyl has a problem in stability compared to the solid form, and it is difficult to control the equivalence depending on the salt and exact molar ratio, and it is difficult to strictly manage the produced acid addition salts . For example, an excess of added organic acid can be precipitated and mixed during the salt formation process, and it is highly unlikely that it will meet the criteria for a drug substance that requires strict management standards.

U.S. Patent Publication No. 2011/0112292 discloses a process for producing a crystalline tennofovirinoceptor free base in order to solve such a problem and discloses a novel crystal form having a melting point of 61 ° C to 66 ° C, . However, such a low melting point has a low thermal stability, and therefore, there are many limitations in its use as an active ingredient of a pharmaceutical drug, and the present invention also relates to the usefulness of crystalline tennofovirinoflorosil as an intermediate for the production of tennofovirinosofroxyl fumarate It emphasizes.

Open Patent Publication No. 2012-0095874 describes a novel salt of tennofovirino soaproxyl and a process for preparing it, and also discloses a process for obtaining a free base from tennofovirino soaproxyl fumarate. In the case of the free base disclosed in the present invention, ethyl acetate is added to the fumaric acid salt, neutralized with sodium bicarbonate, dried and concentrated, and the precipitated solid is obtained by stirring at low temperature. The free base disclosed in the present invention was used as a raw material for producing aspartate which is a crystalline acid addition salt instead of a raw material drug.

The present inventors prepared and analyzed a free base according to the method disclosed in Korean Unexamined Patent Publication No. 2002-0095874. As a result, it was found that a solid having a high tackiness, having a melting point of 94-97 ° C, It is problematic in stability, hygroscopicity and poor purity of the product. Therefore, further purification is required, and the amount of residual solvent after drying conforms to ICH (International Standardization Standard) standard It could not be used as a medicinal raw material in itself.

Accordingly, the present inventors have disclosed a novel crystalline tropopride N-isopropylxyl free base which is thermodynamically stable and has low hygroscopicity and can be stored at room temperature for an extended period of time, has no stickiness and is excellent in solubility, do.

Crystalline acid addition salts of tenofoviruzofoxil have disclosed various crystalline polymorphs in a number of prior arts. However, they are more stable than the conventional crystalline acid addition salts, and have the specifications required by the drug substance and the physico-chemical properties Have not been reported in the prior art or the literature.

When a crystalline free base is used instead of a crystalline acid salt as a pharmaceutical raw material, it is industrially meaningful because it can reduce one step in the production of GMP raw materials. The production method disclosed in the present invention has a very high production yield, It is highly efficient and economical because it has high purity that is not needed.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors have sought to obtain a novel crystalline free base of tennophobia sooperoxil, which overcomes the disadvantages of conventional tennophobia soaploxil fumarate and is superior in physicochemical properties and stability.

As a result, the present inventors have developed methods and conditions for producing novel crystalline forms of tennophobic niobasiloxane free bases having the above-mentioned advantages, and the thus prepared tennofovirino soaproxyl free base is thermodynamically stable, As such, has a high purity suitable for use as an active ingredient of a medicament.

Accordingly, it is an object of the present invention to provide a novel crystalline tropobenzoylphosphoryl free base.

It is a further object of the present invention to provide a process for preparing crystalline tennofovirinoforosol free base.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, the present invention provides a method for determining the specific peak diffraction angles (2 & tilde & 0.1 DEG) in an X-ray diffraction (PXRD) analysis of 7.4 +/- 0.2 DEG, 8.1 +/- 0.2 DEG, 10.8 +/- 0.2 DEG, 14.4 ± 0.2 °, 14.8 ± 0.2 °, 16.2 ± 0.2 °, 16.7 ± 0.2 °, 17.6 ± 0.2 °, 18.04 ± 0.2 °, 19.19 ± 0.2 °, 19.4 ± 0.2 °, 20.9 ± 0.2 °, 22.9 ± 0.2 °, (2), characterized by having a differential scanning calorie (DSC) characteristic of FIG. 2 and a melting point in the range of 107 ± 3 ° C. Lt; RTI ID = 0.0 > sulfopropyl < / RTI >

(2)

The present inventors have sought to obtain a novel crystalline free base of tennophobia sooperoxyl, which overcomes the disadvantages of conventional tennofovirinoforo fosylxyl fumarate and has excellent purity and excellent physico-chemical properties and stability.

As a result, the present inventors have developed methods and conditions for producing novel crystalline forms of tennophobic niobasiloxane free bases having the above-mentioned advantages, and the thus prepared tennofovirino soaproxyl free base is thermodynamically stable, Which is itself suitable for use as an active ingredient in medicines.

The crystalline tropopribisubrozil free base of the present invention represented by formula (2) exhibits an X-ray diffraction pattern different from other known free bases.

According to one embodiment, the crystalline teno Poby where soft lock the free base of the invention has X-ray diffraction pattern shown in Figure 1;

More specifically, the crystalline tropospheric free fluoro silicon free base of the present invention has a differential scanning calorimetric (DSC) characteristic of FIG. 2 and a melting point of 107 3C.

The crystalline tennofovirinoflorosiloxane free base of the present invention can be prepared by reacting a crystalline free base (for example, those disclosed in U.S. Published Patent Application No. 2011/0112292 and Korean Unexamined Patent Publication No. 2002-0095874) and crystalline tannins Compared with povidoserofoxyl fumarate salt, it is excellent in stability, is suitable for the standards of flexible substances and residual solvents, and is a crystalline solid with no hygroscopicity and tackiness, and is suitable for use as an active ingredient of medicines.

According to another aspect of the present invention, the present invention provides a process for preparing a crystalline tropheformed isoproxyl free base comprising the steps of:

(a) dissolving or suspending tennophobic niobasiloxane salt in methylene chloride or a mixed solvent of methylene chloride and methanol, adjusting the pH by adding sodium carbonate or sodium hydrogen carbonate aqueous solution, and separating the organic layer;

(b) adding a desiccant magnesium sulfate (MgSO ₄₎ or sodium sulfate (Na ₂ SO ₄₎ as a result of the step (a) by stirring;

(c) filtering the resultant of the step (b), followed by concentration under reduced pressure and drying;

(d) adding to the result of step (c) ethanol, isopropanol; Or a co-solvent of ethanol and isopropanol, followed by vigorous stirring and selective seeding to obtain a crystalline tropheformed isoproxyl free base.

The manufacturing method of the present invention will be described in detail in each step as follows:

Step (a): Tennophobia norsofloxacin acid salt treatment

In the present invention, the step of dissolving or suspending the tennophobia soapoxylic acid salt in methylene chloride or a mixed solvent of methylene chloride and methanol, adjusting the pH by adding sodium carbonate or an aqueous solution of sodium bicarbonate, and then separating the organic layer by extraction .

On the other hand, the treatment with a sodium carbonate or sodium hydrogen carbonate aqueous solution and methylene chloride or a mixed solvent of methylene chloride and methanol, which are added to the tennophobic niobacillus acid addition salt, are not bound in that order.

The tennofovirinoflorosiloxane salt used in the present invention includes various acid addition salts of tennophobia norsofloxil, for example tennofovirinosofroxyl fumarate.

PH control by treatment with aqueous sodium carbonate or sodium bicarbonate means making the conditions of pH 9-10.

Step (b) - (c): Obtaining crystalline tennofovirinoceptor free base by recrystallization

In step (b), magnesium sulfate (MgSO ₄ ) or sodium sulfate (Na ₂ SO ₄ ) is added as a desiccant and stirred to remove moisture.

Filtering the resultant in step (c), preferably at 5 [deg.] C, and concentrating under reduced pressure and drying at 50 [deg.] C or lower.

Step (d): Obtaining crystalline tennophobic niobasiloxane free base by recrystallization

The resultant product of step (c) or the starting material of step (a) (ii) is added with ethanol or isopropanol and a co-solvent of the two solvents, followed by vigorous stirring and optionally seeding, Lt; / RTI > to afford crystalline tennophobic niobasiloxane free base.

The mixing ratio of ethanol or isopropanol and the two solvents can be varied, and the amount of solvent used is specifically 10-50 ml per 1 g of tennofovirinoceptor free base.

For crystallization, it is advantageous to vigorously agitate the tennophobic niobasiloxane free base and the solvent, and the agitation time is 1-8 hours, more specifically 1-4 hours.

Upon stirring, the precipitation time of crystals can be shortened by selectively seeding the tennophobic niobasiloxane crystalline free base according to the present invention. Seeding is not essential, but it is effective when a crystalline solid is not precipitated even with stirring for several hours.

(a) ethanol, isopropanol in a mucilage free base, an amorphous free base or a crystalline free base of tennophobia isoproxyl; Or a mixed solvent of ethanol and isopropanol (co-solvent);

(b) filtering the result of step (a);

(c) seeding the resultant of step (b) to obtain a crystalline tennofobaranthroxyl free base.

Steps (a) - (b): Treatment of tennophobic nitroso free radical base (mucilage oil or amorphous and crystalline solid)

In this preparation method, a mucinous free base of tennophobia norsofloxacin or an amorphous and other crystalline free base (for example, Korean Unexamined Patent Publication No. 2002-0095874) is prepared in place of tennophobia soaproxylic acid salt as a starting material preparation.

The present invention relates to a process for producing a crystalline tennofovirinoceptor free base using a tennofovir isoproxyl free base as a starting material, which is characterized in that the crystalline tennophobia isoproxyl free base is amorphous or amorphous, As starting materials.

Ethanol, isopropanol, or the like in the crystalline tropospheric soaproxyl free base different from the above-mentioned mucilage oil, amorphous or crystalline form of the present invention; Or a mixed solvent of these two solvents (co-solvent) is added.

Step (c): Obtaining crystalline tennophobic niobasiloxane free base by recrystallization

Crystalline tennophobic niobasiloxane free base is obtained through vigorous stirring and optionally seeding for crystallization of the result of steps (a) - (b).

It is advantageous to vigorously agitate the tennophobic naturally-occurring free base and the solvent, and the agitation time is 1-8 hours, more specifically 1-4 hours.

The features and advantages of the present invention are summarized as follows:

(a) The tennofovirinoflorosilan free base according to the present invention can minimize the generation of a flexible substance according to changes over time compared with the tenofovirnoploxyl fumarate, thereby lowering the amount of impurities generated during the storage of the product, .

(b) The tennofovirisoplosilan free base according to the present invention can also be manufactured with a much higher purity than the known tennophobia soaproxyl free base

(c) In addition, the tennofovirinoceptor free base according to the present invention has physico-chemical properties such as hygroscopicity, solubility and tackiness, and thus can be used as a useful active ingredient of a pharmaceutical composition.

(d) The tennofovirisoplosilan free base according to the present invention is industrially useful since it is suitable for a high yield and high purity, as a raw material for pharmaceutical materials and a residual solvent standard.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a diagram showing an X-ray diffraction pattern of a crystalline tennophobic niobasiloxane free base according to the present invention. FIG.
Figure 2 is a DSC (differential scanning calorimetry) recording of the tennofovir < RTI ID = 0.0 > novolacs free base < / RTI >
FIG. 3 is a graph showing the X-ray diffraction pattern (lower) of the free base prepared by the method described in Korean Patent Laid-Open No. 10-2014-0028790 and the X-ray diffraction pattern of the tennophobic niobasiloxane free base type crystal form of the present invention Respectively.
4 is a graph comparing the DSC data of the tennofovirisoploxil free base crystal form (green graph) of the present invention and the tennofovirinoceptor free base (blue graph) of Korean Patent Publication No. 10-2014-0028790 .
Fig. 5 is a diagram showing the particle size of the tennofovirinoceptor free base of the present invention. Fig.
6 is a diagram showing the particle size of the free base described in Patent Publication No. 10-2014-0028790.
7 is a comparison of powder images of the free base (left) according to the invention disclosed in Korean Patent Laid-Open No. 10-2014-0028790 and the free base of the present invention (right). The free base according to Korean Patent Laid-Open No. 10-2014-0028790 is sticky and lumpy, while the tenofovir's isophosphoryl free base according to the present invention shows a homogeneous powder form.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

In the examples, the 1 H-NMR spectrometer was Agilent 400 MHz, the HPLC was Dionex UltiMate-3000, the melting point was DSC 823 from Mettler Toledo and B-545 from Buchi, Unless otherwise noted, reagents and solvents were purchased from Aldrich.

Example

Example 1: Preparation of a new crystalline form of tennofovirino soaproxyl free base

At room temperature, 200 g of tennophobia iso-roxil fumarate (Asta chemical, China) was dissolved by the addition of 3.6 L of methylene chloride and 360 mL of methanol. 10% sodium carbonate aqueous solution was added thereto, and the pH was adjusted to about 9-10 while stirring for 10 minutes, and then the organic layer was separated. Water was removed from the organic layer using magnesium sulfate, followed by filtration and concentration under reduced pressure at 45 ° C. Thereafter, 600 ml of isopropyl alcohol was added thereto, and the mixture was stirred at 24 ° C for 2 hours and then filtered (washed with 200 ml of isopropyl alcohol) to obtain 140 g of a novel crystalline form of tennophobia soveroxyl base (mp: (Purity): 99.90%).

(1H, d, J = 8.3 Hz), 7.36 (1H, s) 2H), 4.194 (1H, d, J = 6 Hz), 3.999-3.953 (3H, m), 1.246 (12H, d, J = 6H), 1.070 (3H, d, J =

Example 2: Preparation of a new crystalline form of tennofovirino soaproxyl free base

At room temperature, 200 g of tennofovirinosofroxyl fumarate was dissolved in 3.6 L of methylene chloride and 360 mL of methanol. Then, 10% sodium carbonate aqueous solution was added thereto, and the mixture was adjusted to pH 9-10 while stirring for 10 minutes, and then the organic layer was separated. Water was removed from the organic layer using magnesium sulfate, followed by filtration and concentration under reduced pressure at 45 ° C. Thereafter, 300 ml of isopropyl alcohol and 300 ml of ethanol were added, followed by stirring at 24 ° C for 2 hours, followed by filtration (washing with 200 ml of isopropyl alcohol) to obtain 135 g of a new crystalline form of tennophobic niobasiloxane base (mp: , Purity (Purity): 99.90%).

Example 3: Preparation of a new crystalline form of tennofovirino soaproxyl free base

At room temperature, 200 g of tennofovirinosofroxyl fumarate was dissolved in 3.6 L of methylene chloride and 360 mL of methanol. Then, a 10% sodium carbonate aqueous solution was added thereto, and the pH was adjusted to about 9-10 while stirring for 10 minutes, and then the organic layer was separated. Water was removed from the organic layer using magnesium sulfate, followed by filtration and concentration under reduced pressure at 45 ° C. Thereafter, 600 ml of ethanol was added thereto, followed by stirring at 24 ° C for 2 hours and then filtration (washing with 200 ml of isopropyl alcohol) to obtain 135 g of a new crystalline form of tennophobia soveroxyl base (mp: 106 ° C, purity Purity: 99.88%).

Example 4: Preparation of a new crystalline form of tennofovirino soaproxyl free base

At room temperature, 100 g of tennofovirino soaproxylinapadasylate was dissolved by the addition of 1.8 L of methylene chloride and 180 mL of methanol. Then, a 10% sodium carbonate aqueous solution was added thereto, and the pH was adjusted to about 9-10 while stirring for 10 minutes, and then the organic layer was separated. Water was removed from the organic layer using magnesium sulfate, filtered, and concentrated under reduced pressure at 45 ° C. Thereafter, 300 ml of ethanol was added thereto, and the mixture was stirred at 24 ° C for 2 hours and then filtered (washing with 200 ml of isopropyl alcohol) to obtain 56.5 g of a new crystalline form of tennophobia soveroxyl group (melting point: : 99.80%).

Example 5: Preparation of a new crystalline form of tennofovirino soaproxyl free base

400 ml of isopropyl alcohol was added to 100 g of liquid tennophobia soaproxyl free base prepared by the method of US-A-5935946, stirred vigorously at room temperature for 4 hours, and the resulting solid was filtered (isopropyl alcohol (Melting point (mp): 106 캜, purity: 99.89%) as a new crystalline form of tennophobic niobasiloxane free base.

Example 6: Preparation of a new crystalline form of tennofovirino soaproxyl free base

At room temperature, 100 g of tennophobia soaproxylpypidolate was dissolved in 1.8 L of methylene chloride and 180 mL of methanol. Thereafter, the prepared 10% sodium carbonate aqueous solution was added, the pH was adjusted to about 9-10 while stirring for 10 minutes, and the organic layer was separated. Water was removed from the organic layer using magnesium sulfate, followed by filtration and concentration under reduced pressure at 45 ° C. Thereafter, 300 ml of ethanol was added thereto, and the mixture was stirred at 24 ° C for 2 hours and then filtered (washed with 200 ml of isopropyl alcohol) to obtain 61 g of a new crystalline form of tennophobia soveroxyl base (mp: 105.5 ° C, Purity: 99.90%).

Example 7: Preparation of a new crystalline form of tennofovirino soaproxyl free base

300 ml of isopropyl alcohol / 300 ml of ethanol were added to 100 g of the free base obtained by the method disclosed in Korean Patent Publication No. 10-2014-0028790, stirred vigorously at 24 캜 for 4 hours and then filtered (200 ml of isopropyl alcohol (Melting point (mp): 106 占 폚, purity: 99.93%) as a novel crystalline form of tennophobic niobasiloxane base.

Comparative Example 1: Preparation of free base of the method disclosed in Korean Patent Publication No. 10-2014-0028790

125 g of a free base (mp: 96) and a purity of 98.37% were obtained using 200 g of tennofovirinoforosulfosulfur fumarate by a method known in Korean Patent Laid-Open Publication No. 10-2014-0028790. ).

<Experimental Example>

Experimental Example 1: Determination of the crystalline polymorph (XRD) of the novel tennofovirinoceptor free base

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. Thus, the X-ray powder pattern of the tennophobia isoproxyl free base of the present invention was measured.

As a result of the powder X-ray diffraction (PXRD) analysis of the new crystalline form of tennophobivasiloxyl free base, the 2θ diffraction angles were 7.4 ± 0.2 °, 8.1 ± 0.2 °, 10.8 ± 0.2 °, 13 ± 0.2 °, 14.4 ± 0.2 °, 14.8 ± 0.2 °, 16.2 ± 0.2 °, 16.7 ± 0.2 °, 17.6 ± 0.2 °, 18.04 ± 0.2 °, 19.19 ± 0.2 °, 19.4 ± 0.2 °, 20.9 ± 0.2 °, 22.9 ± 0.2 °, 24.4 ± 0.2 °, 26.7 ± 0.2 ° and 28.6 ± 0.2 °, respectively.

Scanning range: 5 to 40 degrees, 2 &thetas;

Scan speed: 6 deg / min

The numerical values of the powder X-ray diffraction (PXRD) angles of the new crystalline form of tennofoibyrofosiloxane free base exhibit diffraction angles different from those of the known tennophobia soaproxyl free base of the previously disclosed U.S. Patent Publication No. 2011/0112292 , And it was confirmed that it is a new crystalline type.

In addition, a diffraction angle pattern different from that of the free base prepared in Korean Patent Laid-Open No. 10-2014-0028790 is shown, and the difference is shown in FIG. 3 as comparative experimental data.

As shown in Fig . 3 , the graph on the upper side is the tennofovir nonosofroxyl free base of the present invention, and the graph on the lower side is free base data described in Korean Patent Laid-Open Publication No. 10-2014-0028790.

Experimental Example 2: Differential Scanning Calorimetry (DSC) analysis of a novel tennofovirino soaproxyl free base

The temperature differential scanning calorimetry (DSC) of the novel crystalline form of tennophobic niobasiloxane free base presented in the present invention for comparison with the novel tennophobia soaproxyl free base having a melting point of 61 ° C to 66 ° C disclosed in US2011 / 0112292 ), The endothermic peak was shown at an endothermic initiation temperature of 103 ± 3 ° C and an endothermic temperature of 107 ± 3 ° C.

Temperature rise range: 20 - 150 ℃

Heating rate: 10 ° C / min

The new crystalline form of tennofovirinoceptor free base proposed in the present invention is thermodynamically stable because it has a melting point of 100 ° C or higher and can be used as an active ingredient of a pharmaceutical product itself, There is also an advantage in producing acid addition salts.

On the other hand, the DSC measurement of the salt prepared in Korean Patent Laid-Open Publication No. 10-2014-0028790 revealed that the endothermic peak was 95 ° C, which was different from the crystal form of the present invention and was not a thermodynamically stable crystalline form.

DSC data of the novel crystalline form of tennofovirinoceptor free base of the present invention (green graph) and tennophobia isoproxyl free base (blue graph) of Korean Patent Laid-Open No. 10-2014-0028790 are shown in FIG . 4 .

Experimental Example 3: Stability Comparative Evaluation (Acceleration / Severe)

The stability test of a drug means to set the validity period by judging the significant change based on the test method after setting the appropriate standard so as to set the storage method and the period of use of the drug etc. Therefore, Is one of the most important factors in the commercialization of drugs.

Therefore, in order to confirm the commercialization of tennophobia soaproxyl free base having the novel crystal form of the present invention, crystalline tennofovir sofloxil fumarate acid (Asta chemicals, china) was used as a control group, and according to the ICH guidelines, Stability and analyzed by liquid chromatography (HPLC) method described in USP (USP). The results are shown in Tables 1 and 2 .

Acceleration (40 ° C ± 2 ° C),
RH 75% ± 5% Flexible material Relative maintenance
time TENOPOVY WHERE SOPROCSYL FUMARICAN New tennophobia isoproxyl free base Patent disclosure publication
The free base of 10-2014-0028790 Early 7 days 30 days Early 7 days 30 days Early 7 days 30 days Tenofovir 0.14 0.002 0.003 0.005 _ _ _ 0.204 0.305 0.405 Adenine 0.16 _ _ _ _ _ _ 0.003 0.006 0.006 TENOPOVY NARROSOCLOSIL MONOESTER 0.24 0.217 0.317 0.356 _ _ _ 0.256 0.776 0.978 Tennophoby isoproxyl ethyl ester 0.80 0.052 0.082 0.231 0.005 0.008 0.010 0.025 0.036 0.046 Tenofovir Soproxil isoester 0.82 _ _ 0.103 _ _ _ 0.011 0.015 0.078 TENOPOVY NARSOPROCSYL CABAMATE 1.40 0.026 0.035 0.037 0.022 0.023 0.029 0.010 0.025 0.036 Tenno Povi Yeroproxil dimer 1.76 _ _ _ _ _ _ _ 0.26 0.28

Severe (60 ° C ± 2 ° C) conditions Flexible material Relative retention time TENOPOVY WHERE SOPROCSYL FUMARICAN New tennophobia isoproxyl free base The free base of Patent Publication No. 10-2014-0028790 Early 7 days 30 days Early 7 days 30 days Early 7 days 30 days Tenofovir 0.14 0.002 0.011 0.105 _ _ _ 0.204 0.305 0.512 Adenine 0.16 _ _ _ _ _ _ 0.003 0.008 0.015 TENOPOVY NARROSOCLOSIL MONOESTER 0.24 0.217 0.335 1.246 _ _ _ 0.256 0.340 0.518 Tennophoby isoproxyl ethyl ester 0.80 0.052 0.105 0.198 0.005 0.005 0.005 0.025 0.105 0.602 Tenofovir Soproxil isoester 0.82 _ _ 0.125 0.001 0.001 0.003 0.011 0.019 0.030 TENOPOVY NARSOPROCSYL CABAMATE 1.40 0.026 0.050 0.053 0.022 0.024 0.024 0.010 0.035 0.356 Tenno Povi Yeroproxil dimer 1.76 _ _ _ _ _ _ _ 0.037 0.056

As shown in Tables 1 and 2 , the tennofovirinoceptor free base having a novel crystal form represented by the present invention was found to be effective as an antioxidant, in comparison with tennophobia soaproxyl fumarate as a control substance under accelerated and severe conditions, It can be seen that it is a very stable substance compared to the free base specified in -2014-0028790. Particularly, it is advantageous that the number and the width of the flexible material are small, so that there is no problem of being affected by degradation products that may occur during storage. In particular, Tenofovirisopropoxylmonoester, which is the main degradation product in tennophobivasiloxyl fumarate acid, is 1.246% in the case of tennofovirinosofroxyl fumarate in harsh conditions, Korean Patent Laid-Open Publication No. 10-2014 For the free bases specified in -0028790 0.518%, respectively. However, the amount of decomposition products of the tennofovirnoploxyl free base prepared in the present invention was 0.1% or less, and the physical and chemical stability was greatly improved.

Therefore, it can be seen that the tennofovirisoplosil free base according to the present invention can maintain a high purity for a long time by minimizing the generation of a flexible substance, and is excellent in storage stability.

Experimental Example 4: Comparative Evaluation of Solubility According to pH

A buffer solution of pH as shown in Table 3 was prepared, and 100 mg of each test substance was stirred at a stirring speed of 25 degrees and 150 rpm for 3 days at each pH, after which each sample was filtered to remove insoluble matter, (HPLC) method.

Based on the solubility obtained under the above conditions, the solubility in terms of pH was determined.

Solubility by pH
Solubility (mg / ml) Tenno Poby Nasofroxil Fumaric acid New free base DW 6.2mg / ml 5.88 mg / ml-equivalent pH 1.2 35.0 mg / ml 34.88 mg / ml-equivalent pH 4.0 4.88 mg / ml 5.12 mg / ml-equivalent pH 6.8 6.45 mg / ml 6.91 mg / ml-equivalent

As shown in Table 3 above, the novel crystalline tennofovirinoflorosilan free base prepared in the present invention has an equivalent degree of solubility in pH buffer solutions as compared to tenofovir soaproxyl fumaric acid. Thus, it can be seen that there is no difference in solubility between the tennofovir isoflocsyl fumaric acid and the tennofovirino soaproxyl free base of the present invention due to the ionization effect.

Experimental Example 5: Hygroscopicity and Particle Size Comparative Evaluation

Since hygroscopicity is one of the important factors for processing and storage among the physical properties required to be used as a raw material drug, the degree of hygroscopicity of the novel crystalline tennofobyrosofloxacin free base of the present invention is compared. Experimental methods were as follows: 500 mg of terfenoviripipifyl fumarate as a control group and 200 mg of the compound of patent publication 10-2014-0028790 500 mg of free base was used and 500 mg of a new crystallized tinofoviruzole free base was placed in a glass tube for hygroscopicity measurement and dried with nitrogen at 25 ° C for 12 hours. After exposure for 12 hours, a hygroscopicity measuring instrument (Model Hydrosorb 1000 ), The mass change rate of the sample was measured automatically at 15, 35, 55, 75, and 95% relative humidity, and is shown in Table 4 .

According to the Korea Food and Drug Administration 's safety standards, tenofovir isofloxyl fumarate is controlled to have a water content of less than 1.0%. As shown in Table 4 , the tenofovir dipivoxil fumarate salt and the free base of Korean Patent Laid-Open Publication No. 10-2014-0028790 rapidly increase in water content as the relative humidity increases, Whereas the soaproxyl free base has relatively low hygroscopicity, it can be regarded as a pharmaceutically stable crystalline form.

Tenno Poby Nasofroxil Fumaric acid New free base Patent disclosure publication
The free base described in < RTI ID = 0.0 > 10-2014-0028790 Remarks Initial mass 0% 0% 0% Relative humidity 15% 0.10% 0% 0% Relative humidity 35% 0.33% 0% 0% Relative humidity 55% 0.56% 0.01% 0.02% Relative humidity 75% 0.80% 0.02% 0.12% Relative humidity 95% 1.42% 0.04% 0.86%

In addition, in the case of solid powder, compatibility with the excipient during tableting, flow rate of the particles, uniformity of the particles, and the like are very important. This is because it is known that the particles are homogeneously distributed on the particle distribution diagram and that the ideal shape powder having a very uniform and symmetrical particle distribution on the PSD is easy to be crushed. The particle size of the free base described in Publication No. 10-2014-0028790 was comparatively measured. Particle size was measured using a conventional solid dispersion method using a mastersizer 2000 instrument. The particle diameters of the novel crystalline tennofovirinoflorosiloxane free base and Patent Document 10-2014-0028790 are shown in Fig . 5 and Fig. 6 , respectively.

As shown in FIG . 5 and FIG. 6 , the novel crystalline tennofovirisoplosil free base of the present invention exhibits a symmetrical particle distribution shape having a narrow and narrow distribution, but the glass of the present invention disclosed in Korean Patent Publication No. 10-2014-0028790 In the case of bases, it has a dispersed morphology of the formulations with a wide variety of peaks.

Further, as shown in Fig . 7 , the free base (left) according to Korean Patent Laid-Open No. 10-2014-0028790 is sticky and lumpy, while the tenofovir < RTI ID = 0.0 > ) Shows a homogeneous powder form.

Claims

In the X-ray diffraction (PXRD) analysis, the specific peak diffraction angles (2? 0.1) were 7.4 ± 0.2 °, 8.1 ± 0.2 °, 10.8 ± 0.2 °, 13 ± 0.2 °, 14.4 ± 0.2 °, 20.9 ± 0.2 °, 22.9 ± 0.2 °, 24.4 ± 0.2 °, 26.7 ± 0.2 °, and 28.6 ± 0.2 °, 16.7 ± 0.2 °, 17.6 ± 0.2 °, 18.04 ± 0.2 °, 19.19 ± 0.2 °, And having a differential scanning calorie value (DSC) characteristic of FIG. 2 and a melting point in the range of 107 占占占 폚. The crystalline tennofobuylsoplosilane free base represented by the following general formula (2)
(2)

2

2. The crystalline base of the tropospheric niobasiloxane according to claim 1, wherein the crystalline tropospheric niobasiloxane free base has an X-ray diffraction pattern as shown in Fig.
1

A process for the preparation of crystalline < RTI ID = 0.0 > tropa-nio < / RTI >
(a) dissolving or suspending tennophobic niobasiloxane salt in methylene chloride or a mixed solvent of methylene chloride and methanol, adjusting the pH by adding sodium carbonate or sodium hydrogen carbonate aqueous solution, and separating the organic layer;
(b) adding a desiccant magnesium sulfate (MgSO ₄₎ or sodium sulfate (Na ₂ SO ₄₎ as a result of the step (a) by stirring;
(c) filtering the resultant of the step (b), followed by concentration under reduced pressure and drying; And
(d) adding to the result of step (c) ethanol, isopropanol; Or a co-solvent of ethanol and isopropanol, followed by vigorous stirring and selective seeding to obtain a crystalline tropheformed isoproxyl free base; And,
The crystalline trophemorpholinosofroxil free base has a specific peak diffraction angle (2? 0.1) of 7.4 ± 0.2 °, 8.1 ± 0.2 °, 10.8 ± 0.2 °, 13 ± 0.2 ° and 14.4 ° in the X-ray diffraction (PXRD) 14.2 ± 0.2, 16.2 ± 0.2, 16.7 ± 0.2, 17.6 ± 0.2, 18.04 ± 0.2, 19.19 ± 0.2, 19.4 ± 0.2, 20.9 ± 0.2, 22.9 ± 0.2, 24.4 ± 0.2 °, 26.7 ± 0.2 ° and 28.6 ± 0.2 °, and has a differential scanning calorie (DSC) characteristic of FIG. 2 and a melting point of 107 ± 3 ° C.
(2)

2

A process for the preparation of crystalline < RTI ID = 0.0 > tropa-nio < / RTI >
(a) ethanol, isopropanol in a mucilage free base, an amorphous free base or a crystalline free base of tennophobia isoproxyl; Or a mixed solvent of ethanol and isopropanol (co-solvent);
(b) filtering the result of step (a);
(c) selectively seeding the resultant of step (b) to obtain a crystalline tennofobaranthroxyl free base; And,
The crystalline trophemorpholinosofroxil free base has a specific peak diffraction angle (2? 0.1) of 7.4 ± 0.2 °, 8.1 ± 0.2 °, 10.8 ± 0.2 °, 13 ± 0.2 ° and 14.4 ° in the X-ray diffraction (PXRD) 14.2 ± 0.2, 16.2 ± 0.2, 16.7 ± 0.2, 17.6 ± 0.2, 18.04 ± 0.2, 19.19 ± 0.2, 19.4 ± 0.2, 20.9 ± 0.2, 22.9 ± 0.2, 24.4 ± 0.2 °, 26.7 ± 0.2 ° and 28.6 ± 0.2 °, and has a differential scanning calorie (DSC) characteristic of FIG. 2 and a melting point of 107 ± 3 ° C.
(2)

2

4. The method according to claim 3, wherein the pH of step (a) is adjusted to a pH range of from 9 to 10.

4. The process of claim 3, wherein the ethanol of step (d), isopropanol; Or a co-solvent of ethanol and isopropanol is used in the range of 10-50 ml per 1 g of GS-free isoproxyl free base.

4. The method according to claim 3, wherein the stirring time for the crystallization of step (d) is 1 to 8 hours.

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