CN114456140A

CN114456140A - Eutectic of icaritin and theophylline

Info

Publication number: CN114456140A
Application number: CN202011238564.0A
Authority: CN
Inventors: 翟立海; 王聚聚; 姚景春; 张明明
Original assignee: Lunan Pharmaceutical Group Corp
Current assignee: Lunan Pharmaceutical Group Corp
Priority date: 2020-11-09
Filing date: 2020-11-09
Publication date: 2022-05-10

Abstract

The invention provides an icariin-theophylline eutectic and a preparation method thereof. The X-ray diffraction spectrum of the eutectic expressed by 2 theta has characteristic peaks at 7.3 +/-0.2 degrees, 9.1 +/-0.2 degrees, 16.0 +/-0.2 degrees and 26.1 +/-0.2 degrees. The preparation method provided by the invention is simple and convenient to operate and is suitable for large-scale application. The prepared eutectic can obviously improve the dissolution characteristic of the icariin and has better stability.

Description

Eutectic of icaritin and theophylline

Technical Field

The invention relates to the field of pharmaceutical co-crystals, in particular to the technical field of icariin organic pharmaceutical co-crystals, and specifically relates to a pharmaceutical co-crystal of icariin and theophylline, and a preparation method and application thereof.

Background

Icaritin (IT) is a polyhydroxy flavonoid monomer component of herba Epimedii of Epimedium of berberidaceae. Pharmacological research shows that the icariin has stronger anti-osteoporosis activity than other flavonoid glycoside compounds in epimedium herb, and has the effects of promoting osteoblast activity and inhibiting osteoclast activity in vitro. The traditional Chinese medicine epimedium has the effects of tonifying kidney and yang, strengthening tendons and bones, dispelling wind and removing dampness, washing sores and killing insects, and relieving fatigue and pain. Icariin, one of the main active ingredients, attracts the attention of numerous scholars at home and abroad in recent years, and deeply and widely researches the pharmacological action of icariin, so far, the results show that the main physiological activity of icariin is to improve the functions of cardiovascular and cerebrovascular systems, enhance the immunity of organisms and regulate endocrine, and simultaneously has the effects of resisting tumor, liver toxicity, hypoxia reoxygenation, bone strengthening and the like.

Although icaritin has a good clinical application prospect, the poor water solubility of icaritin greatly limits the clinical application of icaritin. The solubility of icaritin in water is less than 1 mu g/mL, and the poor water solubility causes the problems of poor oral absorption and low bioavailability. At present, researches on solving the solubility of icariin mainly focus on two aspects, one is to research and prepare a new crystal form of icariin or a derivative thereof, and the other is to improve the solubility of the icariin or the derivative thereof by means of a preparation in a more conventional way. For example, the solubility of a raw material medicine and a physical mixture is remarkably improved by preparing an icariin phospholipid complex (jiadongshi, zhazhili, schfeng and the like. preparation of the icariin phospholipid complex and solid dispersion research [ J ] Chinese herbal medicine, 2010, 9(41)), but the method has the problem that a dissolving medium can damage a medicine structure or evaporate; the solubilizing study of icariin (Jinyan, armor, Zhang Zhenhai and the like) is carried out on icariin by microemulsion technology, and the like, (J, Chinese herbal medicine, 2012, 3(43)) the intestinal absorption characteristic of the icariin self-microemulsion in a Caco-2 cell model is preliminarily studied, although the dissolving and absorption of the icariin can be improved, a large amount of emulsifiers and co-emulsifiers are used in the process, and certain quality safety hazards exist.

However, there are few reports about the research on the raw material form of icariin, and patent CN101200743A reports that crystalline icariin is prepared, but the dissolution property is not improved. Patent CN104860958A reports crystal form A and crystal form B of anhydroicaritin, and research results show that compared with the anhydroicaritin of amorphous powder, the solubility of anhydroicaritin is improved to a certain extent, but the solubility of anhydroicaritin is not improved obviously.

At present, many reports are related to icariin, but the reports mainly relate to the properties of preparation, physicochemical properties, pharmacology and the like, but the solubility of the icariin is not obviously improved. Therefore, a new crystal for remarkably improving the solubility of the icariin still needs to be researched, the physicochemical property of the crystal can be effectively improved, the problem of the solubility of the icariin is fundamentally solved, the addition of auxiliary materials in the production process of the preparation is reduced as much as possible, and the hidden danger of medication is reduced as much as possible.

Disclosure of Invention

In view of the problems of poor solubility of the icariin and the defects of the existing improvement method reported in the prior art, the invention successfully prepares an icariin-theophylline eutectic by a eutectic technology, the eutectic can obviously improve the solubility of the icariin, can obviously improve the solubility and dissolution property of the icariin as an active ingredient of a preparation, and has better physicochemical stability.

In a first aspect of the invention, an icaritin-theophylline co-crystal is provided, wherein the icaritin and theophylline are combined in the following manner in the crystal structure:

preferably, in the co-crystal, the molar ratio of icariin to theophylline is 1:1.

Preferably, the icariin-theophylline eutectic crystal has characteristic peaks at 7.3 +/-0.2 degrees, 9.1 +/-0.2 degrees, 16.0 +/-0.2 degrees and 26.1 +/-0.2 degrees in an X-ray diffraction spectrum expressed by 2 theta by using Cu-Ka radiation.

Preferably, the icariin-theophylline eutectic crystal has characteristic peaks at 7.3 +/-0.2 degrees, 9.1 +/-0.2 degrees, 9.7 +/-0.2 degrees, 14.5 +/-0.2 degrees, 16.0 +/-0.2 degrees, 16.8 +/-0.2 degrees, 26.1 +/-0.2 degrees and 39.8 +/-0.2 degrees by an X-ray diffraction spectrum expressed by 2 theta through Cu-Kalpha radiation.

Preferably, the icariin-theophylline eutectic is irradiated by Cu-Ka, and the characteristic peak of the icariin-theophylline eutectic accords with an X-ray powder diffraction spectrum shown in figure 1.

Preferably, the icariin-theophylline eutectic has an endothermic peak of 225.59 ℃ in a Differential Scanning Calorimetry (DSC) curve.

Preferably, the second aspect of the present invention provides a preparation method of an icariin-theophylline eutectic, which comprises the following specific preparation steps: dissolving icariin and theophylline in an organic solvent, heating and stirring, cooling and crystallizing, filtering and drying to obtain the icariin-theophylline eutectic crystal.

Preferably, the organic solvent is selected from one or more of acetonitrile, acetone, tetrahydrofuran, methanol, ethanol and isopropanol.

Further preferably, the organic solvent is selected from one or two of acetonitrile and acetone.

Preferably, the molar ratio of the icariin to the theophylline is 1: 0.9-1.2; preferably 1: 1.05-1.1.

Preferably, the mass-volume ratio of the icariin to the organic solvent is 5-7: 1, wherein the mass is in mg and the volume is in ml.

Preferably, the heating and stirring time is 2-6 h; preferably 3-4 h.

Preferably, the temperature for dissolving and heating is 45-75 ℃.

Preferably, the temperature reduction and crystallization temperature is 0-15 ℃; preferably 5 to 10 ℃.

Preferably, the crystallization time is 3-5 h.

Further preferably, the preparation method comprises the following steps:

dissolving icariin and theophylline in an organic solvent, heating to dissolve at 45-75 ℃, stirring, refluxing, reacting for 2-6 hours, cooling to 5-10 ℃, crystallizing for 3-5 hours, filtering, washing a filter cake, and drying to obtain the icariin and theophylline eutectic crystal.

Preferably, the solvent for washing the filter cake is selected from one of ethanol, acetonitrile or acetone.

Preferably, the drying temperature is 50-70 ℃, and the drying time is 6-8 hours.

In a third aspect of the invention, a pharmaceutical composition is provided, wherein the composition contains the icariin-theophylline eutectic and is mixed with other components.

Preferably, the pharmaceutical composition of the present invention is prepared as follows: the compounds of the present invention are combined with pharmaceutically acceptable solid or liquid carriers and optionally with pharmaceutically acceptable adjuvants and excipients using standard and conventional techniques to prepare pharmaceutical dosage forms.

Preferably, the other components include pharmaceutically active ingredients, excipients, fillers, etc., which may be used in combination.

Preferably, the pharmaceutical composition is a spray, a tablet, a capsule, a powder injection, an injection and the like.

In the fourth aspect of the invention, the application of the icariin-theophylline eutectic crystal as an active ingredient in preparing a medicament for treating diseases is provided; preferred diseases include various inflammatory diseases, immune diseases, tumors, hyperglycemia, anemia, cardiovascular and cerebrovascular diseases, and the like.

Confirmation of the Crystal Structure

X-ray crystal data were collected on a jtaab Synergy model instrument, japan, testing temperature 293(2) K, irradiating with CuKa, collecting data in an omega scan fashion and Lp correction. The crystal structure is calculated by a ShelXT program in 01ex-2 software, the ShelXL program is adopted to correct structure parameters and judge atom species by a least square method, all hydrogen atom positions are obtained by a geometric calculation method and a difference Fourier method, the degree of similarity (goof value) is 1.019 and is close to 1.0, and the weight scheme is suitable and the structure is accurate.

The crystallographic data obtained by testing and analyzing the icariin-theophylline eutectic prepared by the invention are shown in the table 1, wherein the crystallographic parameters are as follows: triclinic systemThe space group is P-1; the unit cell parameters are:

a is 97.4527(9) °, β is 108.6420(10) °, γ is 114.5138(12) °, unit cell volume

The molecular formula is: c₂₈H₂₈N₄O₈The molecular weight is: 548.54. the ORTEP diagram of the icaritin-theophylline eutectic shows that the icaritin and the theophylline are connected together through intramolecular hydrogen bonds, wherein the hydroxyl H3 of the icaritin and the N3 on the theophylline form the hydrogen bonds, the H4 on the hydroxyl of the icaritin and the keto O5 form the intramolecular hydrogen bonds, and the stacking diagram of the eutectic is shown in figure 4.

TABLE 1 Epimedium aglycone-theophylline eutectic major crystallography data

The X-ray powder diffraction test instrument and the test conditions of the invention are as follows: x-ray powder diffractometer PANalytical E; Cu-K alpha; a sample stage: a flat plate; the incident light path is BBHD; diffraction light path: PLXCEL; voltage 45kv and current 40 mA; a diverging slot 1/4; an anti-scatter slit 1; 0.04rad of cable pull slit; step length: 0.5 s; scanning range: 3 to 50 degrees.

According to the crystallography data, the characteristic peaks in the corresponding X-ray powder diffraction pattern (Cu-Ka) are detailed in attached figure 1 and table 2.

TABLE 2 Main PXRD Peak of icariin-theophylline eutectic

All samples prepared in the examples according to the method of the present invention essentially comply with the X-ray powder diffraction pattern.

The TGA/DSC thermal analysis tester and the test conditions in the invention are as follows: TGA/DSC thermogram METTLER TOLEDO TGA/DSC3 +; dynamic temperature section: 30-300 ℃; heating rate: 10 ℃/min; segment gas N₂(ii) a Gas flow rate: 50 mL/min; crucible: an aluminum crucible of 40. mu.l.

The TGA/DSC test result of the icariin theophylline eutectic prepared by the method is shown in figure 2, and the DSC test result has an endothermic peak with the corresponding temperature of 225.59 ℃. According to the TGA detection result, a weight loss step exists, and calculation shows that the icariin theophylline eutectic does not have solvate.

The method for preparing the icariin-theophylline eutectic crystal provided by the invention is simple and convenient to operate, the prepared crystal is high in purity, the icariin-theophylline eutectic crystal provided by the invention has better stability, the solubility of the icariin is obviously improved, the solubility and dissolution characteristics of the icariin serving as an active ingredient of a preparation can be obviously improved, and a high-quality raw material selection is provided for the production of the preparation of the icariin.

Drawings

FIG. 1: x-ray powder diffraction pattern of icariin-theophylline eutectic.

FIG. 2: DSC-TGA chart of icaritin-theophylline eutectic crystal.

FIG. 3: an ORTEP diagram of the icariin-theophylline eutectic crystal.

FIG. 4: a stacking diagram of the icariin-theophylline eutectic.

Detailed Description

The invention is further illustrated by the following examples, which should be properly understood: the examples of the present invention are intended to be illustrative only and not to be limiting, and therefore, the present invention is intended to be simply modified within the scope of the present invention as claimed.

Example 1

368mg of icariin and 189mg of theophylline are added into a mixed solvent of 30ml of acetone and 40ml of ethanol, the mixture is heated to 50 ℃, stirred, refluxed and reacted for 3 hours, the temperature is slowly reduced to 5-10 ℃, the mixture is kept at a controlled temperature, kept stand and crystallized for 4 hours, filtered, a filter cake is washed by ethanol, and vacuum drying is carried out for 8 hours at 50 ℃ to obtain the icariin and theophylline eutectic crystal, the yield is 90.33%, and the purity is 99.95%.

Example 2

368mg of icariin and 198mg of theophylline are added into a mixed solvent of 30ml of acetonitrile and 40ml of acetone, the mixture is heated to 50 ℃, stirred, refluxed and reacted for 4 hours, the temperature is slowly reduced to 5-10 ℃, the mixture is kept at a controlled temperature, kept stand and crystallized for 4 hours, filtered, a filter cake is washed by acetonitrile, and vacuum drying is carried out for 6 hours at 50 ℃ to obtain the icariin and theophylline eutectic crystal, wherein the yield is 89.68%, and the purity is 99.88%.

Example 3

368mg of icariin and 216mg of theophylline are added into a mixed solvent of 30ml of tetrahydrofuran and 30ml of ethanol, the mixture is heated to 60 ℃, stirred, refluxed and reacted for 5 hours, the temperature is slowly reduced to 5-10 ℃, the mixture is kept at a controlled temperature, kept stand and crystallized for 4 hours, filtered, a filter cake is washed by ethanol, and vacuum drying is carried out for 7 hours at 50 ℃ to obtain the icariin and theophylline eutectic crystal, wherein the yield is 91.25%, and the purity is 99.32%.

Example 4

368mg of icariin and 189mg of theophylline are added into 52ml of acetone, heated to 50 ℃, stirred, refluxed and reacted for 5 hours, slowly cooled to 5-10 ℃, kept at a controlled temperature, kept stand and crystallized for 5 hours, filtered, a filter cake is washed by acetone with the temperature of about 5 ℃, and vacuum-dried for 7 hours at the temperature of 50 ℃ to obtain the icariin and theophylline eutectic crystal, wherein the yield is 87.56%, and the purity is 99.35%.

Example 5

368mg of icariin and 189mg of theophylline are added into 70ml of acetonitrile, the mixture is heated to 50 ℃, stirred, refluxed and reacted for 5 hours, the temperature is slowly reduced to 5-10 ℃, the mixture is subjected to temperature control, standing and crystallization for 4 hours, the filtration is carried out, a filter cake is washed by acetonitrile, and vacuum drying is carried out for 6 hours at 50 ℃ to obtain the icariin and theophylline eutectic crystal, the yield is 87.36%, and the purity is 99.21%.

Stability test

The specific stability test method is carried out according to the guidance method related to stability investigation in the fourth part of the Chinese pharmacopoeia 2015 edition, the purity detection is carried out by an HPLC method, and the specific detection results are shown in Table 3.

TABLE 3 stability test results of icariin-theophylline co-crystal under illumination, high temperature and high humidity conditions

Solubility test

The method comprises the following steps: respectively measuring 10ml of medium (water and 0.01mol/LHCl solution) into penicillin bottles, adding excessive samples to be detected, sealing the penicillin bottles, placing in a constant temperature water bath at 25 ℃, stirring for 1 hour, filtering through a 0.45 mu m filter membrane, and taking filtrate; the absorbance was measured at a wavelength of 270nm, and the solubility was calculated by measuring the absorbance of the standard control.

TABLE 4 solubility of icaritin-theophylline cocrystal in different media

Claims

1. An icaritin-theophylline eutectic crystal is characterized in that the icaritin and the theophylline are combined in the following mode in the crystal structure:

2. the icariin-theophylline co-crystal of claim 1, wherein the molar ratio of icariin to theophylline in the co-crystal is 1:1.

3. The icaritin-theophylline eutectic of claim 1, wherein an X-ray diffraction spectrum expressed in terms of 2 θ using Cu-Ka radiation has characteristic peaks at 7.3 ± 0.2 °, 9.1 ± 0.2 °, 16.0 ± 0.2 °, 26.1 ± 0.2 °.

4. The icaritin-theophylline co-crystal of claim 1, wherein an X-ray diffraction spectrum expressed in terms of 2 θ using Cu-ka radiation has characteristic peaks at 7.3 ± 0.2 °, 9.1 ± 0.2 °, 9.7 ± 0.2 °, 14.5 ± 0.2 °, 16.0 ± 0.2 °, 16.8 ± 0.2 °, 26.1 ± 0.2 °, 39.8 ± 0.2 °.

5. The icariin-theophylline co-crystal of claim 1, wherein the characteristic peak of the icariin-theophylline co-crystal conforms to the X-ray powder diffraction pattern shown in fig. 1 using Cu-ka radiation.

6. The icariin-theophylline co-crystal of claim 1, having crystallographic parameters of: triclinic system, space group is P-1; the unit cell parameters are:

7. A preparation method of icaritin-theophylline eutectic crystal as claimed in any one of claims 1 to 6, comprising the following specific preparation steps: dissolving icariin and theophylline in an organic solvent, heating for dissolving, cooling for crystallization after the solution is clarified, filtering and drying to obtain the icariin-theophylline eutectic crystal.

8. The method according to claim 7, wherein the organic solvent is selected from one or more of acetonitrile, acetone, tetrahydrofuran, methanol, ethanol, and isopropanol.

9. Use of the icariin-theophylline co-crystal as claimed in any one of claims 1 to 6 as an active ingredient in the preparation of a medicament for the treatment of a disease.

10. A pharmaceutical composition comprising the icariin-theophylline co-crystal of any one of claims 1 to 6, and further comprising other ingredients.