CN101671339B - Citric acid alidenafil crystal form B and preparation method and application thereof - Google Patents

Abstract

The invention relates to 1- [3- (6, 7-dihydro-1-methyl-7-oxo-3-propyl-1H-pyrazolo [4, 3-d ] pyrimidine-5-yl) -4-ethoxy benzenesulfonyl ] -cis-3, 5-dimethyl piperazine citrate or citric acid alidenafil crystal form B and a preparation method thereof, and also relates to a pharmaceutical composition containing the citric acid alidenafil crystal form B and application thereof in the preparation of medicaments for treating male Erectile Dysfunction (ED). Proves that an unprecedented citric acid alidenafil crystal form B is obtained, the crystal form B can form a composition with one or more pharmaceutically acceptable carriers, excipients or diluents, and can be effectively applied to the treatment of andropathy.

Description

Aldenafil citrate crystal form B and its preparation method and use

technical field

The present invention relates to 1-[3-(6,7-dihydro-1-methyl-7-oxo-3-propyl-1H-pyrazolo[4,3-d]pyrimidin-5-yl)- Crystal form B of 4-ethoxybenzenesulfonyl]-cis-3,5-dimethylpiperazine citrate (Aildenafil citrate), its preparation method, and the The pharmaceutical composition of the obtained crystal form B and the application of the crystal form B in the manufacture of medicines for treating male erectile dysfunction (male erectile dysfunction, ED) are disclosed.

Background technique

Male erectile dysfunction (male erectile dysfunction, ED) is a common disease, which can be defined as the inability to erect the penis, ejaculate or both. According to statistics, its incidence rate accounts for 1.9% in men over 40 years old, and ED in men over 65 years old For men it reaches 65%. About 125 million men in the world suffer from different degrees of erectile dysfunction, and it is expected to reach 322 million by 2025 (Moreland RB, et al, J Pharmacol Exp Ther, 2001, 296 (2): 225-234.). Therefore, the research and development of safe and effective new drugs or new drug delivery systems for ED has important clinical value and social benefits. For this reason, some new drugs with novel structures and unique mechanisms of action have been introduced to the market or are undergoing clinical and preclinical research.

Aildenafil citrate, chemical name: 1-[3-(6,7-dihydro-1-methyl-7-oxo-3-propyl-1H-pyrazolo[4 ,3-d]pyrimidin-5-yl)-4-ethoxybenzenesulfonyl]-cis-3,5-dimethylpiperazine citrate, molecular formula: C ₂₃ H ₃₂ N ₆ O ₄ S ·C ₆ H ₈ O ₇ , molecular weight: 680.73, chemical structural formula:

Is a new drug in clinical research, effective for ED. The Chinese patent (Application No. 02100198.7) discloses edenafil and its preparation method, etc., but does not involve the crystal form of edenafil citrate and its preparation method.

In the process of studying and preparing Aldenafil citrate, the inventors found that Aldenafil citrate has polymorphism, and there are four crystal forms: Type A, Type B, Type C, and Type D. Type A, Type B, and Type C are crystalline forms without water and other solvents, and Type D is a hydrate. The four crystal forms have high purity and good stability, are superior in industrial production, and are suitable for preparation process and long-term storage.

Contents of the invention

The invention discloses an aldenafil citrate crystal form B, a preparation method of aldenafil citrate crystal form B, a pharmaceutical composition containing aldenafil citrate crystal form B and citric acid Application of Aldenamorph B in the manufacture of drugs for the treatment of male erectile dysfunction (male erectile dysfunction, ED).

The content of the present invention will now be specifically described in conjunction with the purpose of the present invention.

The present invention provides a kind of Aldenafil citrate compound crystal form B, through X-ray crystallographic research, it is characterized in that:

The crystal system and space group are: monoclinic crystal system, the space group is P2(1)/c;

Unit cell parameters: a=13.0384(12) alpha=90deg

b=22.876(2) beta=115.8500(10)deg

gamma＝90deg；c=12.1075(11)

gamma=90deg;

晶胞内分子数＝4，含有4个枸橼酸爱地那非分子；Cell Volume: 3250.0(5)

The number of molecules in the unit cell=4, containing 4 aldenafil citrate molecules;

The calculated crystal density is 1.391 mg/mm ³ ;

Final determined stoichiometric formula: C ₂₉ H ₄₀ N ₆ O ₁₁ S;

Calculated relative molecular mass: 680.73.

Single crystal X-ray diffraction structure analysis is a direct, accurate and most effective crystal form analysis method. It is internationally recognized as the most reliable method for confirming polymorph structure. It can directly obtain crystal unit cell parameters, space groups, crystal The number of molecules in the cell, the three-dimensional structure information of the molecule (including solvent), and then calculate the configuration and conformation of the molecule, the arrangement of the molecule in the crystal, the hydrogen bond, the salt bond and the coordination bond in the molecule and between the molecules.

The detailed crystallographic data of the crystal form B, such as atomic coordinates, angles between bonds (bond angles), and characteristic torsion angles are listed in Table 1-Table 5.

Table 1. Positional parameters of Form B (crystal)

Table 2. Bond lengths and bond angles of Form B (crystal)

Table 3. Anisotropic Position Parameters of Form B (Crystalline)

Table 4. Positional parameters of hydrogen in Form B (crystal)

Table 5. Conformation and characteristic torsion angles of Form B (crystal)

X-ray crystallographic research, especially the crystallographic data in Table 1-Table 5, proves the structure of crystal form B (crystal body), which can be seen intuitively in Figure 1 and Figure 2.

Structural Reliability Factor:

Final R indices[I＞2sigma(I)]

R ₁ =0.0492, WR ₂ =0.1127

R indices(all data)

R ₁ =0.0808, WR ₂ =0.1315

S (Goodness-of-fit on F ² ): 1.042

The edenafil citrate of the crystal form B has a melting point of 212.5-215.0° C. measured by heating the oil bath according to the melting point determination method of the Chinese Pharmacopoeia.

Due to the weak interaction force (such as hydrogen bond, complex bond, etc.) between the molecules inside the unit cell in the crystalline drug, there is a certain difference in the strength of the covalent bond in the molecules of different crystal forms. Infrared absorption spectroscopy (IR) is the result of the energy level transition of the molecular covalent bond, and the difference in the strength of the covalent bond will lead to changes in the IR spectrum. The differences in the infrared spectra of solid drugs with different crystalline forms mainly include changes in peak shape, shift in peak position, and change in peak intensity. Measure with Nicolet 5700 infrared spectrometer, before measuring, calibrate the instrument with polystyrene film, make accord with the regulation of Chinese Pharmacopoeia, use the edenafil citrate of this crystal form B and Potassium bromide (excellent grade pure) mixed pressure during determination. slices, and record the spectra at 4000-400cm ^-1 . The results show that its infrared spectrum at 3580±5cm ^-1 ; 3459±5cm ^-1 ; 3419±5cm ^-1 ; 3266±5cm ^-1 ; 1713±2cm ^-1 has the characteristics that can distinguish it from other crystal forms Absorption peak, see Figure 3. The error range is determined according to the Chinese Pharmacopoeia.

Another object of the present invention discloses the preparation method of edenafil citrate compound crystal form B, the process comprising: adding 25-30 times (weight- Volume ratio, g/ml) of distilled water, add or not add activated carbon accounting for 5% of the weight of Aldenafil citrate, start stirring, heat up to reflux temperature, filter while hot after 15-20 minutes, and seal the filtrate Put it in a filter bottle, place it indoors, cool down naturally, lower it to 25°C-30°C, keep it for 30-48 hours, precipitate crystals, filter, place it indoors for 1-1.5 hours, then move it to a vacuum drying oven and dry it in vacuum for 3 -4 hours, the above crystalline form B of edinamorph citrate was obtained. During the filtrate insulation placement process, it is a necessary condition to stand still and seal the filter bottle holding the filtrate.

Used edenafil citrate is obtained according to the following synthetic route:

Compound 2: 4-amino-1-methyl-3-n-propylpyrazole-5-carboxamide

Compound 3: 2-ethoxybenzoyl chloride

Compound 4: 4-(2-ethoxybenzamide)-1-methyl-3-n-propylpyrazole-5-amide

Compound 5: 1-methyl-3-propyl-5-[(2-ethoxy)phenyl]-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidine-7- ketone

Compound 6: 1-methyl-3-propyl-5-[(2-ethoxy-5-sulfonyl)phenyl]-1,6-dihydro-7H-pyrazolo[4,3-d ]pyrimidin-7-one

Compound 7: cis-2,6-dimethylpiperazine

Compound 8: Aldenafil

Final Compound 1: Aldenafil Citrate

Wherein, compound 2, 3, 7 can be purchased, if can not buy in the market, can be made by the raw material that is easy to obtain by conventional synthetic method according to the precedent of literature, as compound 2 can be according to literature (Chem.Pharm.Bull.1984, 32(4): 1568-1577; Fine Chemical Industry, 2001, 18(7): 396-397, etc.); Compound 3 can be prepared according to the literature (Chemical Research and Application, 2002, 14(5): 605-607, etc.) ; Compound 7 can also be easily prepared according to existing literature methods.

Wherein, compound 4,5,6,8, edenafil citrate, can be according to literature (US4666908; Chinese Journal of Pharmaceutical Industry, 2000,31 (4): 145-147; Chemical Research and Application, 2002, 14( 5): 605-607; Journal of Shenyang Pharmaceutical University, 2002, 19 (3), 174-175. etc.) provides the method for preparing compound 8 from compound 6, only need to use N-methylpiperazine in cis Aldenafil (8) can be conveniently synthesized by replacing formula-2,6-dimethylpiperazine. In order to obtain high-purity Aldenafil citrate, Aldenafil (8) can be recrystallized once with methanol. Then, Aldenafil reacts with equimolar citric acid for 0.5-1 hour in 23-28 times of methanol or ethanol (mass-volume ratio, g/ml) at reflux temperature to generate Aldenafil citrate Non-crude product, recrystallized from methanol, used for crystal form study. Its chemical structure was confirmed by hydrogen nuclear magnetic resonance spectrum ( ¹ H-NMR) and carbon nuclear magnetic resonance spectrum ( ¹³ C-NMR), which proved that the chemical structure was correct, as shown in Fig. 4 and Fig. 5 . In addition, when studying the crystal form, the chemical structure can also be proved to be correct from the single-crystal X-diffraction patterns of the B-type and D-type.

All of the above reactions are routine reactions, and it will be apparent to those skilled in the art that appropriate reagents and conditions for carrying out these reactions can be easily determined by referring to general textbooks and related literature.

Another object of the present invention is to provide a pharmaceutical composition comprising crystalline form B of aldenafil citrate.

Any conventionally known and widely used excipients in the art, such as carriers, fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, lubricants or thinner. Wherein the carrier includes but not limited to lactose, white sugar, sodium chloride, glucose, starch, calcium carbonate, crystalline cellulose and silicic acid. Binders include, but are not limited to, water, ethanol, propanol, dextrose solution, starch solution, gelatin solution, carboxymethylcellulose, methylcellulose, potassium phosphate, and polyvinylpyrrolidone. Disintegrants include, but are not limited to, dry starch, sodium alginate, agar powder, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, monoglyceride stearate, starch, or lactose. Moisturizers include, but are not limited to, glycerin or starch. Lubricants include, but are not limited to, purified talc, stearates, boric acid powder and polyethylene glycols.

The preferred route of administration of the pharmaceutical composition of the present invention is oral. Dosage forms include tablets, granules, capsules, sustained-release tablets, sustained-release pellets and the like. Tablets, granules, capsules are preferred.

The amount of the crystal form edenafil citrate contained in the pharmaceutical composition is 50-70 mg in unit dosage form.

The present invention also provides the application of the aldenafil citrate crystal form B in the manufacture of medicaments for treating male erectile dysfunction.

Pharmacodynamics test: Determination of sexual function in castrated mice

60 Kunming mice (60 female mice with a weight of 18-22 g were fed at the same time) were randomly divided into 6 groups according to body weight, 10 mice in each group, and the bilateral testes of 5 groups were removed under ether anesthesia for castration. The remaining group only underwent surgical separation without removing the testes, and served as the sham operation control group. The animals in each group were returned to the cage for feeding, and the experiment was carried out 3 days later. Both the sham operation control group and the model control group were given 0.5% CMC, the test drug was given aldenafil crystal form B 2, 6, 20 mg.kg ^-1 , and the positive control group was given sildenafil 6 mg.kg ^-1 . For ig administration, the administration volume was 10ml.kg ^-1 . 60 minutes after the administration, the male mice were placed in cages alone, and one female mouse was added to each cage, and the time since the female mice were thrown into the cage for the first time that the male mice caught the female mice (i.e. the capture latency period) was recorded. , and the number of times the male mice climbed their backs within 30 minutes. The results showed that 15mg/kg ^-1 could shorten the capture latency of castrated mice by 180%, and increase the capture times by 3.9 times.

Influencing factor test:

Effect on Appearance

sample project 0 month 0.5 months January Aldenafil citrate compound crystal form B High temperature test High humidity test Lighting test White crystals White crystals White crystals White crystals White crystals White crystals White crystals White crystals White crystals

Effect on content (HPLC area normalization method)

samples project 0 months (%) 0.5 months (%) January(%) Aldenafil citrate compound crystal form B High temperature test High humidity test Lighting test 99.9399.9399.93 99.9299.9399.91 99.9399.9299.93

Effects on Related Substances

sample project 0 months (%) 0.5 months (%) January(%) Aldenafil citrate compound crystal form B High temperature test High humidity test Lighting test 0.070.070.07 0.090.080.08 0.080.080.09

Influence on Infrared Absorption Spectrum

samples project 0 month 0.5 months January Aldenafil citrate compound crystal form B High temperature test High humidity test Lighting test See Figure 3 See Figure 3 See Figure 3 Unchanged Unchanged Unchanged Unchanged Unchanged Unchanged

High temperature, high humidity, light test, 0-1 month, the melting point of Aldina citrate amorphous form B does not change, and the melting point is 212.5-215.0°C.

Results: Aldenafil Citrate Form B was exposed to strong light (4500lx±500lx), high temperature (60±2°C) and high humidity (RH92.5%) from 0 to 1 month, the appearance, melting point, The infrared absorption spectrum has not changed, indicating that the crystal form is stable, no crystal transformation occurs, and the original crystal form is still maintained; in addition, the relevant substances and contents have not changed, indicating that the new crystal form has good chemical stability and is suitable for the manufacture and long-term storage of pharmaceutical preparations .

Attached to the manual:

Fig. 1 is the projection diagram of the molecular three-dimensional structure of aldenafil citrate crystal form B;

Fig. 2 is the unit cell projection figure of aldenafil citrate crystal form B;

Fig. 3 is the infrared spectrogram of aldenafil citrate crystal form B;

Figure 4 is the proton nuclear magnetic resonance spectrum ( ¹ H-NMR) of edenafil citrate;

Fig. 5 is a carbon nuclear magnetic resonance spectrum ( ¹³ C-NMR) of aldenafil citrate.

Detailed ways:

The present invention will be further described below in conjunction with the embodiments and accompanying drawings, so that those skilled in the art can better understand the present invention. The examples are illustrative only and are in no way meant to limit the scope of the invention in any way.

The edenafil citrate used in the present invention has been described above, and its chemical structure is analyzed by elemental analysis, proton nuclear magnetic resonance spectrum ( ¹ H-NMR), carbon nuclear magnetic resonance spectrum ( ¹³ C-NMR, DEPT) 1. Confirmation by high-resolution mass spectrometry (HRMs), which proves that the chemical structure is correct, wherein hydrogen nuclear magnetic resonance spectrum ( ¹ H-NMR) and carbon nuclear magnetic resonance spectrum ( ¹³ C-NMR,) are shown in Figure 4 and Figure 5 .

Example 1

In a 100ml reaction bottle, add 3 grams of edenafil citrate and 88ml of distilled water, start stirring, heat up to the reflux temperature, filter while it is hot after 15 minutes, seal the filtrate in the filter bottle, place it indoors, and cool down naturally , lowered to 25°C-30°C, kept it for 35 hours, precipitated crystals, filtered, placed indoors for 1 hour, then moved to a vacuum drying oven, and dried in vacuum for 3 hours to obtain the above crystal form of aldenamorph citrate B 2.1g, its melting point is m.p 212.5～215.0℃, and the refining rate is 70%. The content measured by HPLC area normalization method is 99.93%. See Fig. 1-Fig. 3, which are the molecular three-dimensional structure projection diagram, unit cell projection diagram and infrared spectrogram of Aldenafil citrate crystal form B, showing the characteristics of Aldenafil crystal form B citrate.

Example 2

In a 250ml reaction bottle, add 5 grams of edenafil citrate, 130ml of distilled water, and 0.25 grams of activated carbon, start stirring, heat up to reflux temperature, and filter while hot after 18 minutes. Seal the filtrate in a filter bottle, place it indoors, lower the temperature naturally to 25°C-30°C, keep it warm for 45 hours, precipitate crystals, filter, place it indoors for 1.5 hours, then move it to a vacuum drying oven and dry it in vacuum for 4 After 1 hour, 3.7 g of the crystalline form B of the edenafil citrate compound was obtained, with a melting point of m.p 212.5 to 215.0° C. and a purification rate of 74%. The content measured by HPLC area normalization method is 99.93%. After testing, it showed the characteristics of edenamorph citrate crystal form B.

Example 3

Granules Containing Aldenafil Citrate Form B

Prescription: 50 grams of Aldenafil Citrate Form B, 650 grams of lactose, 100 grams of crospovidone, 90 grams of PEG-4000, 135 grams of hydroxypropyl methylcellulose, appropriate amount of distilled water, made into 1000 bags.

Process: PEG-4000 and aldenafil citrate crystal form B are crushed together, passed through an 80-mesh sieve, mixed with other materials, made into soft materials with distilled water, granulated, dried at low temperature, and then packed into granules.

Example 4

Capsules containing Aldenafil Citrate Form B

Prescription: 60 grams of Aldenafil citrate crystal form B, 50 grams of starch, 40 grams of lactose, 10 grams of sucrose, 35 grams of microcrystalline cellulose, an appropriate amount of 10% polyvinylpyrrolidone ethanol solution, 1 gram of magnesium stearate, Make 1000 capsules.

Process: Aldenafil citrate crystal form B and auxiliary materials are passed through a 80-mesh sieve, weighed according to the prescription amount, 10% polyvinylpyrrolidone ethanol solution is used as a binder, and a 16-mesh sieve is used to make suitable granules, 65°C Dry, sieve with a 14-mesh sieve, add magnesium stearate and mix evenly, measure the content of the granules, calculate the filling amount, and put it into capsules.

Example 5

Tablets containing Aldenafil Citrate Form B

Prescription: Aldenafil Citrate Form B 70g, Microcrystalline Cellulose 5g, Lactose 140g, PEG-4000 10g, Magnesium Stearate 1g, Povidone K30 14g, Croscarmellose Sodium cellulose 10 grams, appropriate amount of distilled water, made into 1000 tablets.

Process: PEG-4000 and aldenamorph citrate crystal form B are pulverized together, passed through a 80-mesh sieve, mixed with other materials, made into soft materials with distilled water, granulated with a 16-mesh sieve, and placed in a drying oven at 40-45 Dry at ℃, granulate with a 16-mesh sieve, add magnesium stearate to the dry granules, mix evenly, and compress into tablets.

The above content is only a preferred embodiment of the present invention. For those of ordinary skill in the art, according to the idea of the present invention, there will be changes in the specific implementation and application scope. limits.

Claims (7)

1. A crystalline form B of aldenafil citrate, characterized in that: crystal system and space group are: Monoclinic crystal system, the space group is P2(1)/c; Unit cell parameters: a=13.0384(12)

alpha=90deg b=22.876(2) beta=115.8500(10)deg c=12.1075(11)

gamma=90deg; Cell Volume: 3250.0(5)

The number of molecules in the unit cell=4, containing 4 aldenafil citrate molecules; The calculated crystal density is 1.391 mg/mm ³ ; Final determined stoichiometric formula: C ₂₉ H ₄₀ N ₆ O ₁₁ S; Calculated relative molecular mass: 680.73. 2. The crystalline form B of aldenafil citrate according to claim 1, characterized in that: the infrared spectrum of the crystalline form B is at 3580±5cm ^-1 , 3459±5cm ^-1 , 3419±5cm ^{-1 1.} There are characteristic peaks at 3266±5cm ^-1 and 1713±2cm ^-1 . 3. The crystalline form B of edenafil citrate according to claim 1, characterized in that its melting point is 212.5-215.0°C. 4. the preparation method of edenafil citrate crystalline form B described in claim 1 is characterized in that: edenafil citrate is dissolved in 25-30 times (mass-volume ratio, g/ml) In boiling distilled water, the temperature was lowered to 25°C-30°C, airtight, heat-preserved and left to stand for 30-48 hours, and crystals were precipitated to obtain the above crystalline form B of aldenamorph citrate. 5. A use of edenafil citrate crystalline form B according to any one of claims 1 to 3, characterized in that: edenafil citrate crystalline form B is combined with one or more pharmaceutically Acceptable carriers and excipients make up the composition. 6. The use of the crystalline form B of edenafil citrate according to claim 5, characterized in that: the composition is used to prepare oral preparations. 7. The use of edenafil citrate crystalline form B according to claim 5, characterized in that: the application of edenafil citrate crystalline form B in the manufacture of medicines for treating male erectile dysfunction.

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