Patchouli oil-cyclodextrin inclusion compound co-crystal and its pharmaceutical composition and application

Abstract

The present application relates to the field of pharmaceutical preparations and fine chemical technology, and specifically to a patchouli oil-cyclodextrin inclusion complex eutectic and its pharmaceutical composition and application. The patchouli oil-cyclodextrin inclusion complex eutectic is an inclusion complex eutectic formed by co-crystallization of the cyclodextrin and the patchouli oil after the patchouli oil is encapsulated with cyclodextrin; the mass ratio of the cyclodextrin to the patchouli oil is (2-30):1. Compared with traditional inclusion complexes, the patchouli oil-cyclodextrin inclusion complex eutectic of the present application has a clear crystal structure and better stability, and the efficacy of the treatment of ulcerative colitis is also significantly improved.

Description

Technical Field

[0001] The present application relates to the field of pharmaceutical preparations and fine chemical technology, and in particular to a patchouli oil-cyclodextrin inclusion complex eutectic and its pharmaceutical composition and application. Background Art

[0002] Patchouli is a commonly used Chinese medicine for the clinical treatment of digestive system diseases in traditional Chinese medicine, and it has a long history of use. Patchouli oil is the main effective part of Patchouli, which is included in the 2020 edition of the Chinese Pharmacopoeia. It has pharmacological effects such as the treatment of gastrointestinal inflammatory diseases and is a component of the prescription of Huoxiang Zhengqi Water, a major Chinese medicine variety. Patchouli oil is a volatile liquid substance with poor stability and high transportation difficulty. This characteristic hinders its further application in drug development and clinical practice.

[0003] Existing methods for improving the stability of volatile oil drugs mainly include the use of new formulation technologies such as soft capsules, microcapsules, nanotechnology, etc. for physical isolation - this method usually requires more special production equipment, which may increase production costs, and the use of pharmaceutical excipients such as silicon dioxide, PEG, etc. for adsorption and solidification to improve the stability and water solubility of the drug - this method may have potential effects on drug release and bioavailability, and it is also necessary to ensure the safety and stability of all additives and carrier materials.

[0004] In view of this, it is necessary to provide a patchouli oil pharmaceutical preparation that is simple to prepare, safe and stable. Summary of the invention

[0005] Based on this, one or more embodiments of the present application provide a patchouli oil-cyclodextrin inclusion compound co-crystal and its pharmaceutical composition and application. The patchouli oil-cyclodextrin inclusion compound of the present application has high safety, good stability and simple preparation method.

[0006] The technical solution of this application includes the following contents:

[0007] A patchouli oil-cyclodextrin inclusion compound eutectic comprises patchouli oil and cyclodextrin wrapped on the surface of the patchouli oil in its structure, and the mass ratio of the cyclodextrin to the patchouli oil is (2-30):1.

[0008] Optionally, the cyclodextrin is β-cyclodextrin or γ-cyclodextrin;

[0009] When the cyclodextrin is β-cyclodextrin, the X-ray powder diffraction pattern of the patchouli oil-cyclodextrin inclusion complex cocrystal has characteristic diffraction peaks at the following 2θ (°) angles: 5.7°±0.2°, 6.0°±0.2°, 6.6°±0.2°, 7.2°±0.2°, 9.9°±0.2°, 11.5°±0.2°, 12.2°±0.2°, 12.7°±0.2°, 14.2°±0.2°, 15.8°±0.2°, 16.1°±0.2°, 17.3°±0.2°, 18.9°±0.2°, 19.4°±0.2°, 20.8°±0.2°, 21.3°±0.2°, 22.4°±0.2°, 23.9°±0.2°, 24.8°±0.2°, 25.9°±0.2°, 26.8°±0.2°, 27.9°±0.2°, 28. .2°, 14.7°±0.2°, 15.6°±0.2°, 16.3°±0.2°, 17.2°±0.2°, 17.6°±0.2°, 18.8°±0.2°, 19.7°±0.2°, 21.3°±0.2°, 22.5°±0.2°, 23.0°±0.2°, 23.4°±0.2°, 24.0°±0.2°, 26.0°±0.2°;

[0010] When the cyclodextrin is γ-cyclodextrin, the X-ray powder diffraction pattern of the patchouli oil-cyclodextrin inclusion complex cocrystal has characteristic diffraction peaks at the following 2θ (°) angles: 5.5°±0.2°, 7.6°±0.2°, 9.4°±0.2°, 10.7°±0.2°, 12.0°±0.2°, 13.8°±0.2°, 14.4°±0.2°, 15.9°±0.2°, 16.0°±0.2°, 17.1°±0.2°, 18.8°±0.2°, 19.4°±0.2°, 20. 5.1°±0.2°, 16.8°±0.2°, 17.7°±0.2°, 19.4°±0.2°, 20.5°±0.2°, 21.3°±0.2°, 22.0°±0.2°, 23.8°±0.2°, 24.3°±0.2°, 25.6°±0.2°, 26.9°±0.2°, 27.6°±0.2°.

[0011] A method for preparing the patchouli oil-cyclodextrin inclusion complex co-crystal as described above comprises the following steps:

[0012] Dissolving patchouli oil in an organic solvent to obtain a patchouli oil solution, wherein the organic solvent is miscible with water;

[0013] Under stirring and heating conditions, injecting the patchouli oil solution into the cyclodextrin aqueous solution to obtain a mixed solution, wherein the amount of injection is such that the mixed solution contains cyclodextrin and patchouli oil in a mass ratio of (2-30):1;

[0014] The mixed solution was allowed to stand for crystallization.

[0015] Optionally, the organic solvent comprises at least one of methanol, ethanol, acetonitrile, acetone, tetrahydrofuran, isopropanol and ethylene glycol; and / or

[0016] The cyclodextrin is β-cyclodextrin or γ-cyclodextrin.

[0017] Furthermore, the preparation method satisfies one or more of the following conditions:

[0018] (1) The concentration of the patchouli oil solution is 0.1 mg / mL to 10 mg / mL;

[0019] (2) The concentration of the cyclodextrin aqueous solution is 3 mg / mL to 100 mg / mL;

[0020] (3) The stirring speed is 50 rpm to 1000 rpm;

[0021] (4) The heating temperature is 30°C to 60°C;

[0022] (5) The rate of injecting the patchouli oil solution into the cyclodextrin aqueous solution is 0.5 to 2 mL / min.

[0023] (6) In the mixed solution, the volume ratio of the organic solvent to water is 2:(3-40).

[0024] Optionally, when the aqueous solution of cyclodextrin is an aqueous solution of β-cyclodextrin, the concentration is 3 mg / mL to 20 mg / mL;

[0025] When the aqueous solution of cyclodextrin is an aqueous solution of γ-cyclodextrin, the concentration is 5 mg / mL to 100 mg / mL.

[0026] Further, the mixed solution is allowed to stand for crystallization:

[0027] The standing is performed at 4°C to 30°C; and / or

[0028] The standing time is 2h to 48h.

[0029] Further, the mixed solution is allowed to stand, after crystallization, the solid is collected and dried;

[0030] Optionally, the drying temperature is 30°C to 60°C;

[0031] Optionally, the drying time is 1 hour to 48 hours.

[0032] A pharmaceutical composition comprising:

[0033] 1) the patchouli oil-cyclodextrin inclusion compound co-crystal described above, and

[0034] 2) Pharmaceutically acceptable carriers and / or excipients.

[0035] A use of the patchouli oil-cyclodextrin inclusion complex co-crystal described above in preparing a drug for treating ulcerative colitis.

[0036] The present application uses cyclodextrin to encapsulate patchouli oil, and allows cyclodextrin and patchouli oil to co-crystallize to form inclusion complex co-crystals. Compared with traditional inclusion complexes, the patchouli oil-cyclodextrin inclusion complex co-crystals of the present application have a clear crystal structure and better stability. Unlike conventional co-crystal preparation methods that utilize hydrogen bonds, electrostatic effects, etc. between drugs and co-crystal ligands to form crystals, the inclusion complex co-crystals utilize the characteristic that cyclodextrin and patchouli oil can form inclusion complexes. The two participate in crystal formation in the form of inclusion complexes, which increases the water solubility of patchouli oil on the one hand, and increases the stability of patchouli oil on the other hand. At the same time, the efficacy of the patchouli oil-cyclodextrin inclusion complex co-crystals of the present application in treating ulcerative colitis is also significantly improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 This is an optical microscope photo of the white powder obtained in Examples 1 and 2 of the present application;

[0038] Figure 2 H-NMR spectra of the white powder, patchouli oil, β-cyclodextrin, and γ-cyclodextrin obtained in Examples 1 and 2 of the present application, wherein the abscissa represents chemical shift (ppm) and the ordinate represents intensity;

[0039] Figure 3 The infrared spectra of the white powder, patchouli oil, β-cyclodextrin and γ-cyclodextrin prepared in Examples 1 and 2 of the present application, wherein the abscissa is the wave number (cm -1 ), the ordinate represents absorbance;

[0040] Figure 4 The PXRD spectra of the white powder, β-cyclodextrin, and γ-cyclodextrin prepared in Examples 1 and 2 of the present application, wherein the abscissa represents the angle 2θ (°) and the ordinate represents the intensity;

[0041] Figure 5 The comparison results of the patchouli oil preparations of Example 1 and Comparative Examples 1-2 of the present application and the storage stability of patchouli oil at 60°C are shown;

[0042] Figure 6 The comparison results of the in vitro drug release of the products of Example 1 and Comparative Examples 1-2 of the present application and patchouli oil;

[0043] Figure 7 These are the results of the efficacy evaluation of the patchouli oil-cyclodextrin inclusion complex co-crystal in the treatment of acute colitis according to the examples of the present application. DETAILED DESCRIPTION

[0044] In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the relevant drawings. The preferred embodiments of the present invention are given in the drawings. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0045] The words "preferably", "more preferably", etc. in the present invention refer to embodiments of the present invention that may provide certain beneficial effects in certain circumstances. However, other embodiments may also be preferred under the same circumstances or other circumstances. In addition, the description of one or more preferred embodiments does not imply that other embodiments are not applicable, nor is it intended to exclude other embodiments from the scope of the present invention.

[0046] When a numerical range is disclosed herein, the above range is considered to be continuous and includes the minimum and maximum values ​​of the range, as well as every value between such minimum and maximum values. Further, when a range refers to an integer, every integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe features or characteristics, the ranges can be combined. In other words, unless otherwise indicated, all ranges disclosed herein should be understood to include any and all subranges included therein.

[0047] Unless otherwise specified, all percentages, fractions and ratios are calculated by the total mass of the composition of the present invention. Unless otherwise specified, all masses of the listed ingredients are given to the content of active substances, so they do not include solvents or by-products that may be contained in commercially available materials. The term "mass percentage content" herein can be represented by the symbol "%". Unless otherwise specified, all molecular weights in this article are weight average molecular weights expressed in Daltons. Unless otherwise specified, all formulations and tests in this article occur in an environment of 25°C. "Including", "including", "containing", "containing", "having" or other variants herein are intended to cover non-closed inclusions, and these terms are not distinguished. The term "comprising" means that other steps and ingredients that do not affect the final result can be added. The composition and method / process of the present invention include, consist of and are essentially composed of the necessary elements and restrictions described herein and any additional or optional ingredients, components, steps or restrictions described herein. The terms "efficacy", "performance", "effect" and "efficacy" are not distinguished between herein.

[0048] "Pharmaceutically acceptable" refers to those ligands, materials, compositions, and / or dosage forms that are suitable for administration to a patient within the scope of sound medical judgment and commensurate with a reasonable benefit / risk ratio.

[0049] "Pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. As used herein, the language "pharmaceutically acceptable carrier" includes buffers, sterile water for injection, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are compatible with drug administration. Each carrier must be "pharmaceutically acceptable" in the sense of being compatible with the other ingredients in the formulation and not harmful to the patient. Suitable examples include, but are not limited to: (1) sugars such as lactose, glucose and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) 10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffered saline; and (21) other non-toxic compatible substances used in pharmaceutical formulations.

[0050] As used herein, "drug" includes any agent, compound, composition or mixture that provides physiological and / or pharmacological effects in vivo or in vitro, and often provides beneficial effects. The scope of the physiological and / or pharmacological effects produced by the "drug" in vivo is not particularly limited, and may be systemic effects or may only produce effects locally. The activity of the "drug" is not particularly limited, and may be an active substance that can interact with other substances, or an inert substance that does not interact.

[0051] The dosage form and administration method of the compound of the present invention or its pharmaceutical composition are not particularly limited.

[0052] Representative routes of administration include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous) injection, and topical administration.

[0053] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or solubilizers, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain a buffer. Solid dosage forms such as tablets, sugar pills, capsules, pills and granules may be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions may be delayed in a certain part of the digestive tract. Examples of embedding components that may be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the above-mentioned excipients.

[0054] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, specifically, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances. In addition to these inert diluents, the composition may also contain adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and spices. For example, the suspension may contain a suspending agent, specifically, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitan esters, microcrystalline cellulose, aluminum methylate and agar or mixtures of these substances.

[0055] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous or non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0056] Dosage forms for topical administration include ointments, powders, patches, sprays and inhalants, which are prepared by mixing the active ingredient with a pharmaceutically acceptable carrier and any preservatives, buffers, or propellants that may be required under sterile conditions.

[0057] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0058] In one aspect of the present application, a patchouli oil-cyclodextrin inclusion complex co-crystal is provided, wherein the structure of the inclusion complex co-crystal includes patchouli oil and cyclodextrin wrapped on the surface of patchouli oil, and the mass ratio of cyclodextrin to patchouli oil is (2-30):1.

[0059] Optionally, the cyclodextrin is β-cyclodextrin or γ-cyclodextrin;

[0060] When the cyclodextrin is β-cyclodextrin, the X-ray powder diffraction pattern of the patchouli oil-cyclodextrin inclusion complex cocrystal has characteristic diffraction peaks at the following 2θ (°) angles: 5.7°±0.2°, 6.0°±0.2°, 6.6°±0.2°, 7.2°±0.2°, 9.9°±0.2°, 11.5°±0.2°, 12.2°±0.2°, 12.7°±0.2°, 14.2°±0.2 °, 14.7°±0.2°, 15.6°±0.2°, 16.3°±0.2°, 17.2°±0.2°, 17.6°±0.2°, 18.8°±0.2°, 19.7°±0.2°, 21.3°±0.2°, 22.5°±0.2°, 23.0°±0.2°, 23.4°±0.2°, 24.0°±0.2°, 26.0°±0.2°;

[0061] When the cyclodextrin is γ-cyclodextrin, the X-ray powder diffraction pattern of the patchouli oil-cyclodextrin inclusion complex cocrystal has characteristic diffraction peaks at the following 2θ (°) angles: 5.5°±0.2°, 7.6°±0.2°, 9.4°±0.2°, 10.7°±0.2°, 12.0°±0.2°, 13.8°±0.2°, 14.4°±0.2°, 15. 1°±0.2°, 16.8°±0.2°, 17.7°±0.2°, 19.4°±0.2°, 20.5°±0.2°, 21.3°±0.2°, 22.0°±0.2°, 23.8°±0.2°, 24.3°±0.2°, 25.6°±0.2°, 26.9°±0.2°, 27.6°±0.2°.

[0062] In yet another aspect of the present application, a method for preparing patchouli oil-cyclodextrin inclusion compound co-crystal is provided. This preparation method can prepare the patchouli oil-cyclodextrin inclusion compound co-crystal mentioned above.

[0063] The preparation method of the present application combines the process principles of preparing nanoparticles by solvent injection method and crystallization by anti-solvent method. The key lies in: 1) selecting an organic solvent miscible with water as a good solvent for patchouli oil; 2) preparing patchouli oil-cyclodextrin inclusion complex eutectic by solvent injection method.

[0064] This preparation method comprises the following steps:

[0065] S100: dissolving patchouli oil in an organic solvent to obtain a patchouli oil solution, wherein the organic solvent is miscible with water;

[0066] S200: injecting a patchouli oil solution into a cyclodextrin aqueous solution under stirring and heating conditions to obtain a mixed solution, wherein the amount of the injected solution is such that the mixed solution contains cyclodextrin and patchouli oil in a mass ratio of (2 to 30):1;

[0067] S300: The mixed solution is allowed to stand for crystallization.

[0068] On the one hand, the organic solvent is a good solvent for patchouli oil and can be used as a diluent and transfer medium for patchouli oil. When the organic solution of patchouli oil is injected into the aqueous solution of cyclodextrin, the organic solvent can quickly combine with water molecules, thereby promoting the dispersion of patchouli oil in water and the formation of inclusion complexes. On the other hand, the organic solvent can be used as an anti-solvent for cyclodextrin to promote the precipitation of the inclusion complex from the solution.

[0069] Optionally, the organic solvent is selected from methanol, ethanol, acetonitrile, acetone, tetrahydrofuran, isopropanol, ethylene glycol, and combinations thereof.

[0070] The patchouli oil solution has a suitable concentration to facilitate dispersion. Optionally, the concentration of the patchouli oil solution is 0.1 mg / mL to 10 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL, 5 mg / mL, 10 mg / mL, etc.

[0071] The cyclodextrin may be selected from β-cyclodextrin or γ-cyclodextrin as required.

[0072] Optionally, the concentration of the cyclodextrin aqueous solution is 3 mg / mL to 100 mg / mL, for example 3 mg / mL, 10 mg / mL, 20 mg / mL, 50 mg / mL, 80 mg / mL, 100 mg / mL, etc.

[0073] Furthermore, when the aqueous solution of cyclodextrin is an aqueous solution of β-cyclodextrin, the concentration is 3 mg / mL to 20 mg / mL, for example 3 mg / mL, 5 mg / mL, 10 mg / mL, 15 mg / mL, 20 mg / mL, etc.;

[0074] When the aqueous solution of cyclodextrin is an aqueous solution of γ-cyclodextrin, the concentration is 5 mg / mL to 100 mg / mL, and further 10 mg / mL to 30 mg / mL, for example 5 mg / mL, 10 mg / mL, 20 mg / mL, 30 mg / mL, 50 mg / mL, 100 mg / mL, etc.

[0075] In S200, stirring and heating not only promote the dissolution of cyclodextrin, but also improve the dispersion effect of patchouli oil in the aqueous solution of cyclodextrin, thereby affecting the coating result and the subsequent crystallization result. Therefore, it is necessary to carry out the process under appropriate stirring and heating.

[0076] Optionally, the stirring speed is 50 rpm to 1000 rpm. This speed is conducive to the formation of a suitable crystal inclusion compound in combination with the other parameters of step S200. Further, the stirring speed can be selected from 100 rpm to 1000 rpm, for example, 100 rpm, 200 rpm, 300 rpm, 400 rpm, 500 rpm, 600 rpm, 700 rpm, 800 rpm, 900 rpm, 1000 rpm, etc.

[0077] Optionally, the heating temperature is 30° C. to 60° C. This temperature, combined with other parameters of step S200 , is conducive to the formation of inclusion complexes.

[0078] Optionally, the speed of injecting the patchouli oil solution into the cyclodextrin aqueous solution is 0.5-2 mL / min. This speed, combined with the other parameters of step S200, is conducive to the formation of inclusion complexes of suitable crystal forms.

[0079] Optionally, the speed of injecting the patchouli oil solution into the aqueous solution of cyclodextrin is 0.5-2 mL / min, and examples of the speeds include 0.5 mL / min, 1 mL / min, 1.5 mL / min, 2 mL / min, etc.

[0080] In S200, a patchouli oil solution is injected into an aqueous solution of cyclodextrin to obtain a mixed solution. The amount of injection is sufficient to make the mixed solution contain cyclodextrin and patchouli oil in a mass ratio of (2 to 30):1. Injecting a suitable amount of patchouli oil into the aqueous solution of cyclodextrin is for better dispersion and obtaining a desired crystalline inclusion complex.

[0081] In some embodiments, the mass ratio of cyclodextrin to patchouli oil in the mixed solution is (2-30):1, for example 2:1, 5:1, 10:1, 12:1, 15:1, 18:1, 20:1, 25:1, 30:1, etc.

[0082] Optionally, in the mixed solution, the volume ratio of the organic solvent to water is 2:(3-40).

[0083] To obtain the above-mentioned preferred concentration and volume ratio, after the patchouli oil solution is completely injected, an organic solvent may be optionally added. The organic solvent is preferably the organic solvent used to prepare the patchouli oil solution in step S100.

[0084] Further, the mixed solution is allowed to stand for crystallization:

[0085] Standing at 4°C to 30°C; and / or

[0086] The standing time is 2h to 48h.

[0087] Further, the mixed solution is allowed to stand, after crystallization, the solid is collected and dried;

[0088] Optionally, the drying temperature is 30°C to 60°C;

[0089] Optionally, the drying time is 1 hour to 48 hours.

[0090] In another aspect of the present application, a pharmaceutical composition is provided, comprising:

[0091] 1) the patchouli oil-cyclodextrin inclusion complex co-crystal described in any of the above technical solutions, and

[0092] 2) Pharmaceutically acceptable carriers and / or excipients.

[0093] In another aspect of the present application, there is provided a use of the patchouli oil-cyclodextrin inclusion complex co-crystal described in any of the above technical solutions in the preparation of a drug for treating ulcerative colitis.

[0094] The present application is further described in detail below in conjunction with specific examples. The raw materials used in the following examples, unless otherwise specified, are all commercially available products.

[0095] 1. Preparation

[0096] Example 1

[0097] This example provides a patchouli oil-cyclodextrin inclusion complex eutectic and its preparation process. The cyclodextrin raw material used is β-cyclodextrin. The specific steps are as follows:

[0098] 1) preparing a β-cyclodextrin aqueous solution and a patchouli oil solution respectively, wherein the solvent of the patchouli oil is ethanol, the concentration of the patchouli oil solution is 2 mg / mL, and the concentration of the β-cyclodextrin aqueous solution is 10 mg / mL.

[0099] 2) The cyclodextrin aqueous solution was heated to 60° C., and the patchouli oil solution was injected into 20 mL of the cyclodextrin aqueous solution at a rate of 1 mL / min under stirring at 1000 rpm, and stirring was continued for 5 minutes to obtain a mixed solution.

[0100] 3) Stop heating and stirring, cool the mixed solution to 20°C, and place it at 4°C for 24 hours until the crystals in the solution are completely precipitated.

[0101] 4) The white solid was filtered off and washed with ethanol for three times. The product was then dried at 60° C. for 24 hours to obtain a white powdery product. The yield of the product based on patchouli oil was 85%.

[0102] Example 2

[0103] This example provides a patchouli oil-cyclodextrin inclusion complex eutectic and its preparation process. The cyclodextrin raw material used is γ-cyclodextrin. The specific steps are as follows:

[0104] 1) preparing a γ-cyclodextrin aqueous solution and a patchouli oil solution respectively, wherein the solvent of the patchouli oil is ethanol, the concentration of the patchouli oil solution is 5 mg / mL, and the concentration of the γ-cyclodextrin aqueous solution is 30 mg / mL.

[0105] 2) The cyclodextrin aqueous solution was heated to 50° C., and the patchouli oil solution was injected into 7 mL of the cyclodextrin aqueous solution at a rate of 0.5 mL / min under stirring at 50 rpm. 2 mL of ethanol was added and stirring was continued for 3 minutes to obtain a mixed solution.

[0106] 3) Stop heating and stirring, cool the mixed solution to 25°C, and leave it for 12 hours until the crystals in the solution are completely precipitated.

[0107] 4) The white solid was filtered off and washed with methanol for 3 times. The product was then dried under vacuum at 40° C. for 12 hours to obtain a white powdery product. The yield of the product based on patchouli oil was 90%.

[0108] Comparative Example 1

[0109] This comparative example provides a patchouli oil product obtained by solidifying patchouli oil using mesoporous silica adsorption technology, and the preparation steps are as follows:

[0110] Preparation of patchouli oil adsorbed on silica: Patchouli oil (80 mg) was dissolved in 200 μL of ethanol, and then 300 mg of mesoporous silica was added to the solution. The resulting mixture was stirred for 5 h, and the resulting product was collected by centrifugation, washed twice with 5% ethanol, and then dried at 50°C under vacuum for 6 h.

[0111] Comparative Example 2

[0112] This comparative example provides a patchouli oil product obtained by solidifying patchouli oil using inclusion technology, and the preparation steps are as follows:

[0113] Weigh about 4g of β-cyclodextrin, add appropriate amount of water and heat to dissolve to make a saturated solution. Keep this saturated solution at 45℃. Take 1mL of patchouli oil and slowly drip it into the saturated solution of β-cyclodextrin, stir at constant temperature for 2h, take it out and cool it, put it in the refrigerator for 24h, take it out and filter it, wash it with a little anhydrous ethanol 3 times. Dry the powder in a vacuum dryer for 24h.

[0114] 2. Characterization

[0115] The white powder products of Examples 1 and 2 were characterized respectively, including morphology, H-NMR, infrared spectrum and X-ray powder diffraction (XRD), and finally confirmed that the products prepared in Examples 1 and 2 were all patchouli oil-cyclodextrin inclusion compound eutectics. The characterization results are as follows:

[0116] 2.1 Morphology observation

[0117] The white powders prepared in Examples 1 and 2 were respectively placed on glass slides, and the morphology of each sample was observed and recorded under an optical microscope (10×20). The results are as follows: Figure 1 shown.

[0118] Figure 1 It shows that the white powders obtained in Examples 1 and 2 have clear crystal forms, which are block-shaped (white powder obtained in Example 1) and long rod-shaped (white powder obtained in Example 2). Under the same experimental conditions, cyclodextrin does not precipitate solids in the solvent. This shows that the white powders obtained in Examples 1-2 are crystalline.

[0119] 2.2H-NMR characterization

[0120] Patchouli oil, β-cyclodextrin, γ-cyclodextrin, and the white powders prepared in Examples 1 and 2 were dissolved in DMSO-d6 and subjected to H NMR spectrum detection. The test results were as follows: Figure 2The H-NMR data of the white powder of Example 1 respectively contain the signals of patchouli oil and β-cyclodextrin, and the H-NMR data of the white powder of Example 2 contain the signals of patchouli oil and γ-cyclodextrin, indicating that the white powder prepared in Example 1 or 2 is composed of patchouli oil and cyclodextrin, confirming that the white powder prepared in Examples 1 and 2 is a eutectic.

[0121] 2.3 Infrared characterization

[0122] Patchouli oil, β-cyclodextrin, γ-cyclodextrin, and the white powders obtained in Examples 1 and 2 were subjected to infrared spectroscopy tests. The test results were as follows: Figure 3 . Figure 3 The characteristic peak of patchouli oil is at 2922cm -1 、2867cm -1 、1641cm -1 、1557cm -1 、1451cm -1 、1374cm -1 、1278cm -1 、1056cm -1 、1038cm -1 、999cm -1 、884cm -1 、787cm -1 、532cm -1 ; The characteristic peak of cyclodextrin is at 400cm -1 ~2000cm -1 Compared with the above two examples, most of the characteristic peaks of patchouli oil in the infrared spectrum of the white powder obtained in Examples 1 and 2 disappeared, and its infrared spectrum was consistent with cyclodextrin (corresponding to β-cyclodextrin and γ-cyclodextrin, respectively), indicating that patchouli oil and cyclodextrin formed an inclusion complex, so the characteristic peaks of patchouli oil in the structure were covered by cyclodextrin. This shows that the white powder obtained in Examples 1 and 2 is an inclusion complex.

[0123] 2.4 XRD characterization

[0124] Patchouli oil, β-cyclodextrin, γ-cyclodextrin, and the white powders obtained in Examples 1 and 2 were subjected to PXRD analysis. The analysis results are as follows: Figure 4 .

[0125] The X-ray powder diffraction pattern of the white powder of Example 1 has characteristic diffraction peaks at the following 2θ (°) angles: 5.7°±0.2°, 6.0°±0.2°, 6.6°±0.2°, 7.2°±0.2°, 9.9°±0.2°, 11.5°±0.2°, 12.2°±0.2°, 12.7°±0.2°, 14.2°±0.2°, 14.7°±0.2°. 0.2°, 15.6°±0.2°, 16.3°±0.2°, 17.2°±0.2°, 17.6°±0.2°, 18.8°±0.2°, 19.7°±0.2°, 21.3°±0.2°, 22.5°±0.2°, 23.0°±0.2°, 23.4°±0.2°, 24.0°±0.2°, 26.0°±0.2°;

[0126] The X-ray powder diffraction pattern of the white powder of Example 2 has characteristic diffraction peaks at the following 2θ (°) angles: 5.5°±0.2°, 7.6°±0.2°, 9.4°±0.2°, 10.7°±0.2°, 12.0°±0.2°, 13.8°±0.2°, 14.4°±0.2°, 15.1°±0.2°, 16.8°±0.2°, 17.7°±0.2°, 19.4°±0.2°, 20.5°±0.2°, 21.3°±0.2°, 22.0°±0.2°, 23.8°±0.2°, 24.3°±0.2°, 25.6°±0.2°, 26.9°±0.2°, and 27.6°±0.2°.

[0127] By comparison, it can be seen that the XRD diffraction peaks of the inclusion complex co-crystals of each embodiment are completely different from the characteristic peaks of each cyclodextrin and patchouli oil, indicating that it is not a simple physical mixture, but a new crystal form is formed, and the HNMR results show that the crystal contains both cyclodextrin and patchouli oil components, so it can be determined to be a cyclodextrin inclusion complex co-crystal of patchouli oil.

[0128] 3. Eutectic performance evaluation

[0129] 3.1 Accelerated stability test

[0130] Patchouli oil, the patchouli oil-cyclodextrin inclusion compound eutectic of Example 1, and the patchouli oil products of Comparative Examples 1-2 were subjected to an accelerated stability test at 60°C. The test results are as follows: Figure 5 .

[0131] according to Figure 5It can be seen that patchouli oil completely volatilizes within one week, the patchouli oil-cyclodextrin inclusion compound eutectic of Example 1 volatilizes within 20% within one week, the patchouli oil inclusion compound (patchouli oil product of Comparative Example 2) and the patchouli oil adsorbed on mesoporous silica (patchouli oil product of Comparative Example 1) volatilize by 40% and more than 70%, respectively. It can be seen that compared with patchouli oil alone or its conventional preparation, the thermal stability of the patchouli oil-cyclodextrin inclusion compound eutectic of the embodiment of the present application is greatly improved.

[0132] 3.2 In vitro dissolution test

[0133] Patchouli oil, the patchouli oil-cyclodextrin inclusion compound co-crystal of Example 1, and the patchouli oil product of Comparative Example 1 were subjected to in vitro dissolution experiments in a simulated gastrointestinal fluid environment. The experimental results are as follows: Figure 6 .

[0134] according to Figure 6 It can be seen that patchouli oil is completely released in gastric juice within half an hour, and the patchouli oil-cyclodextrin inclusion compound co-crystal of Example 1 releases 70% in gastric juice within 2 hours and 30% in intestinal juice within 1 hour, indicating that the patchouli oil-cyclodextrin inclusion compound co-crystal of the present application has a certain sustained-release effect.

[0135] 3.3 Pharmacodynamic experiments

[0136] The dextran sulfate sodium (DSS)-induced ulcerative colitis (IBD) mouse model was used to investigate the therapeutic effect of patchouli oil on intestinal inflammation. The steps are as follows:

[0137] 3.3.1 Establishment of acute IBD model

[0138] Female C57 mice were divided into 5 groups, with 6 mice in each group, namely blank control group, model group, positive drug treatment group, free patchouli oil treatment group, and patchouli oil-cyclodextrin inclusion complex cocrystal treatment group.

[0139] The entire modeling cycle was 10 days, of which: the blank control group was not treated and drank normal sterile water. The mice in the model group, positive drug treatment group, free patchouli oil treatment group, and patchouli oil-cyclodextrin inclusion complex cocrystal treatment group began to drink sterile water containing 2.5% DSS on day 0, and were replaced with normal drinking water on day 8.

[0140] In addition, drug treatment was started on day 3. The positive drug treatment group was treated with mesalazine at a dose of 30 mg / kg, and the free patchouli oil treatment group and the patchouli oil-cyclodextrin inclusion complex cocrystal treatment group were treated with 50 mg / kg (white powder of Example 1) orally.

[0141] All mice were sacrificed on day 10.

[0142] 3.3.2 Calculation of disease activity index

[0143] During the experiment, the body weight change rate was recorded daily, stool characteristics were observed, and blood in stool was measured to calculate the disease activity index.

[0144] 3.3.3 Tissue sample processing

[0145] When the animal experiment reached the end point, the mice were euthanized and dissected. The colon and cecum tissues of the mice were collected, and the length of the colon was recorded and weighed.

[0146] 3.4 Experimental Results

[0147] Figure 7 A is the weight change rate of mice in each group. Figure 7 B is the disease activity index (DAI) score of each group of mice. From the weight of each group of mice and their DAI scores, it can be seen that patchouli oil inclusion complex cocrystal can significantly reduce loose stools and bloody stools in IBD mice. The therapeutic effect of the patchouli oil inclusion complex cocrystal treatment group on the ulcerative colitis mouse model is equivalent to that of the positive drug sulfasalazine (SASP), and there is no significant difference in the efficacy of the free patchouli oil group compared with the model group.

[0148] Figure 7 Middle C is the colon length of mice in each group at the end point of treatment; Figure 7 D is the ratio of colon mass to length of each group of mice. The colon length and mass of each group of mice increased to varying degrees compared with the model group, and the colon length of the PAO eutectic group was significantly higher than that of the PAO group and the model group. The above efficacy results show that the efficacy of PAO volatile oil is significantly improved after solidification into inclusion eutectic.

[0149] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0150] The above-described embodiments only express several implementation methods of the present invention, which are convenient for understanding the technical solutions of the present invention in detail, but they cannot be understood as limiting the scope of protection of the invention patent. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. It should be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided by the present invention are all within the protection scope of the claims attached to the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the contents of the attached claims, and the description and drawings can be used to interpret the contents of the claims.

Claims

1. A patchouli oil-cyclodextrin inclusion compound eutectic, characterized in that: The structure of the inclusion compound eutectic comprises patchouli oil and cyclodextrin coated on the surface of the patchouli oil, and the mass ratio of the cyclodextrin to the patchouli oil is (2-30):

1.

2. The patchouli oil-cyclodextrin inclusion compound co-crystal according to claim 1, characterized in that: The cyclodextrin is β-cyclodextrin or γ-cyclodextrin; When the cyclodextrin is β-cyclodextrin, the X-ray powder diffraction pattern of the patchouli oil-cyclodextrin inclusion complex cocrystal has characteristic diffraction peaks at the following 2θ (°) angles: 5.7°±0.2°, 6.0°±0.2°, 6.6°±0.2°, 7.2°±0.2°, 9.9°±0.2°, 11.5°±0.2°, 12.2°±0.2°, 12.7°±0.2°, 14.2°±0.2°, 15.8°±0.2°, 16.1°±0.2°, 17.3°±0.2°, 18.9°±0.2°, 19.4°±0.2°, 20.8°±0.2°, 21.3°±0.2°, 22.4°±0.2°, 23.9°±0.2°, 24.8°±0.2°, 25.9°±0.2°, 26.8°±0.2°, 27.9°±0.2°, 28. .2°, 14.7°±0.2°, 15.6°±0.2°, 16.3°±0.2°, 17.2°±0.2°, 17.6°±0.2°, 18.8°±0.2°, 19.7°±0.2°, 21.3°±0.2°, 22.5°±0.2°, 23.0°±0.2°, 23.4°±0.2°, 24.0°±0.2°, 26.0°±0.2°; When the cyclodextrin is γ-cyclodextrin, the X-ray powder diffraction pattern of the patchouli oil-cyclodextrin inclusion complex cocrystal has characteristic diffraction peaks at the following 2θ (°) angles: 5.5°±0.2°, 7.6°±0.2°, 9.4°±0.2°, 10.7°±0.2°, 12.0°±0.2°, 13.8°±0.2°, 14.4°±0.2°, 15.9°±0.2°, 16.0°±0.2°, 17.1°±0.2°, 18.8°±0.2°, 19.4°±0.2°, 20. 5.1°±0.2°, 16.8°±0.2°, 17.7°±0.2°, 19.4°±0.2°, 20.5°±0.2°, 21.3°±0.2°, 22.0°±0.2°, 23.8°±0.2°, 24.3°±0.2°, 25.6°±0.2°, 26.9°±0.2°, 27.6°±0.2°.

3. A method for preparing the patchouli oil-cyclodextrin inclusion complex cocrystal according to claim 1 or 2, characterized in that: The steps include: Dissolving patchouli oil in an organic solvent to obtain a patchouli oil solution, wherein the organic solvent is miscible with water; Under stirring and heating conditions, injecting the patchouli oil solution into the cyclodextrin aqueous solution to obtain a mixed solution, wherein the amount of injection is such that the mixed solution contains cyclodextrin and patchouli oil in a mass ratio of (2-30):1; The mixed solution was allowed to stand for crystallization.

4. The preparation method according to claim 3, characterized in that: The organic solvent comprises at least one of methanol, ethanol, acetonitrile, acetone, tetrahydrofuran, isopropanol and ethylene glycol; and / or The cyclodextrin is β-cyclodextrin or γ-cyclodextrin.

5. The preparation method according to claim 3 or 4, characterized in that: The preparation method meets one or more of the following conditions: (1) The concentration of the patchouli oil solution is 0.1 mg / mL to 10 mg / mL; (2) The concentration of the cyclodextrin aqueous solution is 3 mg / mL to 100 mg / mL; (3) The stirring speed is 50 rpm to 1000 rpm; (4) The heating temperature is 30°C to 60°C; (5) injecting the patchouli oil solution into the cyclodextrin aqueous solution at a rate of 0.5 to 2 mL / min; (6) In the mixed solution, the volume ratio of the organic solvent to water is 2:(3-40).

6. The preparation method according to claim 5, characterized in that: When the aqueous solution of cyclodextrin is an aqueous solution of β-cyclodextrin, the concentration is 3 mg / mL to 20 mg / mL; When the aqueous solution of cyclodextrin is an aqueous solution of γ-cyclodextrin, the concentration is 5 mg / mL to 100 mg / mL.

7. The preparation method according to claim 3 or 4, characterized in that: The mixed solution is allowed to stand for crystallization: The standing is performed at 4°C to 30°C; and / or The standing time is 2h to 48h.

8. The preparation method according to claim 3 or 4, characterized in that: The mixed solution is allowed to stand for crystallization, and then the solid is collected and dried; Optionally, the drying temperature is 30°C to 60°C; Optionally, the drying time is 1 hour to 48 hours.

9. A pharmaceutical composition, characterized in that It includes: 1) The patchouli oil-cyclodextrin inclusion compound co-crystal according to claim 1 or 2, and 2) Pharmaceutically acceptable carriers and / or excipients.

10. Use of the patchouli oil-cyclodextrin inclusion complex co-crystal according to claim 1 or 2 in the preparation of a medicament for treating ulcerative colitis.

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