Drug-drug salt nanocrystals, preparation and application thereof

Abstract

The application provides a drug-drug salt nanocrystal of vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen and a preparation and application thereof. The obtained nanocrystal can realize synergistic effect of vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen, significantly enhances the antidepressant activity, realizes the slow-release treatment effect, and thus achieves the purpose of long-acting treatment of depression.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a drug-drug salt nanocrystal, its preparation method, a pharmaceutical composition containing the same, and its applications. Background Technology

[0002] Depression is a common mental health disorder characterized by persistent sadness and anhedonia. This illness imposes a significant socioeconomic burden, severely impairs quality of life, and is associated with significant morbidity and mortality. However, mainstream antidepressants have limitations such as poor efficacy and the need for frequent dosing. Drug-drug salts are an emerging pharmaceutical technology that combines two therapeutically active drugs (usually an acid and a base) through a salt-forming process, forming a single molecular entity with dual pharmacological activities. This strategy enhances therapeutic efficacy by improving the physicochemical properties of the drugs and also increases therapeutic effect through synergistic mechanisms.

[0003] In recent years, nanocrystal technology has played an important role in long-acting injection systems, with significant advantages including effectively reducing dosing frequency, maintaining stable plasma drug concentrations, and prolonging the duration of action. For example, abilify, a once-monthly injectable drug approved for maintenance treatment of schizophrenia... Currently, nanocrystal technology continues to receive widespread attention in the development of novel formulations, demonstrating promising prospects.

[0004] Vortioxetine is a novel antidepressant developed by Lundbeck Pharmaceuticals of Denmark and Takeda Pharmaceutical Company of Japan. It received FDA approval in September 2013 under the brand name Brintellix. This drug acts as a 5-HT3, 5-HT7, and 5-HT1D receptor antagonist, a partial agonist of the 5-HT1B receptor, a agonist of the 5-HT1A receptor, and a 5-HT transporter inhibitor. It exerts its antidepressant effect by regulating 5-HT levels and is used to treat major depressive disorder in adults. The chemical name of vortioxetine is 1-[2-(2,4-dimethylphenylthio)phenyl]piperazine, and its structure is shown in formula (I).

[0005]

[0006] Currently, vortioxetine, with its multi-target mechanism of action, has shown efficacy in relieving depression, improving anxiety, and improving sleep. However, common gastrointestinal side effects and the need for frequent medication often lead to low patient adherence. Recent studies have confirmed that neuroinflammation is one of the key mechanisms in the pathogenesis of depression, with the core process being the activation of microglia, which triggers a cascade of pro-inflammatory cytokines. Nonsteroidal anti-inflammatory drugs (NSAIDs) can exert their anti-inflammatory effects by inhibiting cyclooxygenase and reducing the production of pro-inflammatory mediators such as prostaglandins. Examples include diclofenac and (S)-flurbiprofen, whose structural formulas are shown below.

[0007] Summary of the Invention

[0008] Objective of the Invention: This invention aims to overcome the shortcomings of existing technologies and provide a drug-drug salt nanocrystal of vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen. This drug-drug salt nanocrystal can achieve a synergistic effect of vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen, enhancing antidepressant activity while achieving a sustained-release effect, reducing the frequency of dosing, and fundamentally improving patient treatment adherence. Furthermore, the preparation process of this invention is simple, reproducible, and suitable for industrial production.

[0009] To achieve the above-mentioned objectives, the present invention adopts the following technical solution:

[0010] In a first aspect, the present invention provides a drug-drug salt nanocrystal, characterized in that: the nanocrystal contains a) a drug-drug salt of vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen; and b) a stabilizer.

[0011] As one implementation scheme, the drug-drug salt nanocrystals are vortioxetine-diclofenac drug-drug salt nanocrystals.

[0012] As one embodiment, the nanocrystals are vortioxetine-diclofenac drug-drug salt nanocrystals, and the powder X-ray diffraction pattern of the nanocrystals includes the following characteristic peaks at 2θ angles: 5.656°, 7.859°, 11.179°, 13.481°, 14.381°, 16.781°, 18.521°, 19.240°, 20.178°, 20.860°, 22.359°, 23.080°, 25.077°, 28.883°, and 33.400°, with a 2θ value accuracy of ±0.2°;

[0013] As one embodiment, the powder X-ray diffraction pattern of the vortioxetine-diclofenac drug-drug salt nanocrystals, expressed in terms of diffraction angle 2θ, is essentially as follows: Figure 3 As shown.

[0014] As one embodiment, the nanocrystals are vortioxetine-diclofenac drug-drug salt nanocrystals, and the differential scanning calorimetry curve of the nanocrystals has an endothermic peak at 185.3±1℃.

[0015] As one implementation scheme, the DSC spectrum of the vortioxetine-diclofenac drug-drug salt nanocrystals is basically as follows: Figure 2 As shown (VOT-DIC nanocrystal group).

[0016] As one embodiment, the nanocrystals are vortioxetine-diclofenac drug-drug salt nanocrystals, the nanocrystals having a particle size of 100-500 nm and a dispersibility index of 0.000-0.300.

[0017] As one embodiment, the vortioxetine-diclofenac drug-drug salt nanocrystals are prepared using vortioxetine-diclofenac drug-drug salt as raw material, wherein the molar ratio of vortioxetine to diclofenac is approximately 1:1;

[0018] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, which are prepared from vortioxetine-diclofenac drug-drug salt crystals. The drug-drug salt crystals are monoclinic with space group P21 / C and cell parameters as follows: α = 90°, β = 102.073(3)°, γ = 90°, and the unit cell volume is

[0019] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, which are prepared from vortioxetine-diclofenac drug-drug salt crystals. The single-crystal structure of the vortioxetine-diclofenac drug-drug salt is basically as shown in the figure. Figure 1 As shown.

[0020] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, which are prepared using vortioxetine-diclofenac drug-drug salt crystals as raw materials. The powder X-ray diffraction pattern of the drug-drug salt crystals includes the following characteristic peaks at 2θ angles: 5.663°, 7.877°, 11.193°, 13.501°, 14.418°, 16.794°, 18.546°, 19.285°, 20.179°, 20.878°, 22.068°, 22.370°, 23.113°, 25.094°, 28.016°, and 33.371°, with an accuracy of ±0.2° for the 2θ values.

[0021] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, which are prepared from vortioxetine-diclofenac drug-drug salt crystals. The powder X-ray diffraction pattern of the vortioxetine-diclofenac drug-drug salt crystals, expressed in terms of diffraction angle 2θ, is essentially as follows: Figure 3 As shown.

[0022] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, prepared from vortioxetine-diclofenac drug-drug salt as raw material. The infrared absorption spectrum of the vortioxetine and diclofenac drug-drug salt crystals, measured using KBr pellets, is at 3433.62 cm⁻¹. -1 1634.70cm -1 1578.16cm -1 1558.55cm -1 1469.58cm -1 1450.78cm -1 1373.43cm -1 1315.38cm -1 1230.18cm -1 1151.23cm -1 1044.82cm -1 813.78cm -1 746.51cm -1 There is an absorption peak at this point.

[0023] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, which are prepared from vortioxetine-diclofenac drug-drug salt as raw material, wherein the differential scanning calorimetry curve of vortioxetine and diclofenac drug-drug salt crystals has an endothermic peak at 190.5±1℃.

[0024] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, which are prepared using vortioxetine-diclofenac drug-drug salt crystals as raw materials. The DSC spectrum of the vortioxetine-diclofenac drug-drug salt crystals is essentially as follows: Figure 2 As shown (VOT-DIC group).

[0025] As one implementation scheme, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, which are prepared from vortioxetine-diclofenac drug-drug salt as raw material. The thermogravimetric analysis curve of the vortioxetine and diclofenac drug-drug salt crystals shows that weight loss begins at 240.9±1℃, corresponding to the melting and decomposition process of the vortioxetine-diclofenac drug-drug salt crystals.

[0026] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, which are prepared from vortioxetine-diclofenac drug-drug salt crystals. The TG spectrum of the vortioxetine-diclofenac drug-drug salt crystals is approximately as shown in the figure. Figure 5 As shown.

[0027] As one embodiment, the drug-drug salt nanocrystals are vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals.

[0028] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine-(S)-flurbiprofen, and the powder X-ray diffraction pattern of the nanocrystals includes the following characteristic peaks at 2θ angles: 5.600°, 9.159°, 11.100°, 13.276°, 15.624°, 16.202°, ​​18.200°, 19.641°, 20.120°, 22.280°, 23.476°, 23.999°, 25.201°, 26.201°, 28.619°, and 31.740°, with a 2θ value accuracy of ±0.2°;

[0029] As one embodiment, the powder X-ray diffraction pattern of the vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals, expressed in terms of diffraction angle 2θ, is essentially as follows: Figure 13 As shown.

[0030] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine-(S)-flurbiprofen, and the differential scanning calorimetry curve of the nanocrystals shows endothermic activity at 122.7±1℃.

[0031] As one implementation scheme, the DSC spectrum of the vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals is basically as follows: Figure 12 As shown (VOT-SFLU nanocrystal group).

[0032] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine-(S)-flurbiprofen, and the nanocrystals have a particle size of 100-500 nm and a dispersibility index of 0.000-0.300.

[0033] As one embodiment, the vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals are prepared using vortioxetine-(S)-flurbiprofen drug-drug salt as raw material, wherein the molar ratio of vortioxetine to (S)-flurbiprofen is approximately 1:1;

[0034] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, prepared from vortioxetine-(S)-flurbiprofen drug-drug salt crystals. The vortioxetine and (S)-flurbiprofen drug-drug salt crystals are monoclinic with space group P212121 and cell parameters as follows: α = 90°, β = 90°, γ = 90°, cell volume is

[0035] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, which are prepared from vortioxetine-(S)-flurbiprofen drug-drug salt crystals. The single-crystal structure diagram of the vortioxetine and (S)-flurbiprofen drug-drug salt crystals is basically as shown below. Figure 11 As shown.

[0036] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, which are prepared from vortioxetine-(S)-flurbiprofen drug-drug salt crystals. The powder X-ray diffraction pattern of the vortioxetine and (S)-flurbiprofen drug-drug salt crystals includes the following characteristic peaks at 2θ angles: 5.527°, 9.129°, 11.053°, 13.249°, 15.628°, 16.186°, 18.176°, 19.630°, 20.156°, 22.223°, 23.428°, 23.996°, 25.174°, 26.112°, 28.752°, and 31.442°, with an accuracy of ±0.2° for the 2θ values.

[0037] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, prepared from vortioxetine-(S)-flurbiprofen drug-drug salt crystals. The powder X-ray diffraction pattern of the vortioxetine and (S)-flurbiprofen drug-drug salt crystals, expressed in terms of diffraction angle 2θ, is essentially as follows: Figure 13 As shown.

[0038] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, prepared from vortioxetine-(S)-flurbiprofen drug-drug salt. The infrared absorption spectrum of the vortioxetine and (S)-flurbiprofen drug-drug salt crystals, measured using KBr pellets, is at 3434.65 cm⁻¹. -1 1620.78cm -1 1580.05cm -1 1471.23cm -1 1383.22cm -1 1351.34cm -1 1229.42cm -1 1125.53cm -1 1044.85cm -1 926.74cm -1 873.08cm -1 766.84cm -1 696.48cm -1 There is an absorption peak at this point.

[0039] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, which are prepared from vortioxetine-(S)-flurbiprofen drug-drug salt as raw material, wherein the differential scanning calorimetry curve of vortioxetine and (S)-flurbiprofen drug-drug salt crystals has an endothermic peak at 127.0±1℃.

[0040] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, which are prepared from vortioxetine-(S)-flurbiprofen drug-drug salt as raw materials. The DSC spectra of the vortioxetine and (S)-flurbiprofen drug-drug salt crystals are essentially as follows: Figure 12 As shown (VOT-SFLU group).

[0041] As one implementation scheme, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, which are prepared from vortioxetine-(S)-flurbiprofen drug-drug salt. The thermogravimetric analysis curve of the vortioxetine and (S)-flurbiprofen drug-drug salt crystals shows that weight loss begins at 249.3±1℃, corresponding to the melting and decomposition process of the vortioxetine-(S)-flurbiprofen drug-drug salt crystals.

[0042] As one embodiment, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, which are prepared from vortioxetine-(S)-flurbiprofen drug-drug salt as raw materials. The TG curves of the vortioxetine and (S)-flurbiprofen drug-drug salt crystals are essentially as follows: Figure 15 As shown.

[0043] Secondly, the present invention provides a method for preparing the above-mentioned raw material drug-drug salt, which is a solution method.

[0044] As a specific implementation plan, the solution method includes the following steps:

[0045] 1) Add vortioxetine to an organic solvent and mix to obtain system A;

[0046] 2) Add diclofenac or (S)-flurbiprofen to an organic solvent and mix to obtain system B;

[0047] 3) Mix system A and system B, then add organic solvent or water to obtain system C, and let it stand;

[0048] 4) Filter, wash, collect filter cake, and dry to obtain the drug-drug salt of vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen;

[0049] As one implementation, in steps 1-2), the organic solvent is selected from n-propanol or acetone.

[0050] As one implementation, in step 3), the organic solvent is selected from one or more of ketone solvents, alcohol solvents, alkyl solvents, haloalkyl solvents, alkyl sulfoxide solvents, or cyclic ether solvents.

[0051] As one embodiment, in step 3), the organic solvent is selected from one or more of the following: C1-C6 ketone solvents, C1-C6 alcohol solvents, C1-C6 alkyl solvents, C1-C6 haloalkyl solvents, C1-C6 alkyl sulfoxide solvents, or C1-C6 cyclic ether solvents.

[0052] As one implementation, in step 3), the organic solvent is selected from one or more of acetone, methanol, ethanol, isopropanol, chloroform, n-hexane, ethyl acetate, or dimethyl sulfoxide.

[0053] As one implementation scheme, in step 3), the organic solvent is selected from n-hexane.

[0054] As one implementation scheme, in step 4), the total mass ratio of vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen to the total volume ratio of the solvent in system C is 29-30 mg / ml.

[0055] As one implementation scheme, in step 4), the molar ratio of vortioxetine to diclofenac or vortioxetine to (S)-flurbiprofen is 1:1.

[0056] As one implementation scheme, in step 4), the standing conditions are: standing at 10-28℃ for 12-36 hours.

[0057] Thirdly, the present invention provides a method for preparing the above-mentioned drug-drug salt nanocrystals, wherein the preparation method is wet grinding.

[0058] As one embodiment, the method for preparing vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen drug-drug salt nanocrystals is characterized in that: the nanocrystals contain vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen drug-drug salt, stabilizer and water;

[0059] As one embodiment, the stabilizer is selected from any one or more of hydroxypropyl methylcellulose, povidone, poloxamer, sodium carboxymethyl cellulose, Tween 80, and Tween 20. Preferably, the stabilizer is selected from any one or more of hydroxypropyl methylcellulose, poloxamer, and Tween 80. More preferably, the stabilizer is selected from Tween 80.

[0060] As one implementation, the weight ratio of the nanocrystals to the stabilizer is (1-3):(0.5-2).

[0061] As one embodiment, the preparation method of the vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen drug-drug salt nanocrystals includes the following steps: mixing vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen drug-drug salt, Tween 80 and water, wherein the mass ratio of the drug-drug salt to Tween 80 is (1-3):(0.5-2), wet milling the resulting mixture at 600 rpm for 10 minutes using a ball mill to obtain vortioxetine and diclofenac or (S)-flurbiprofen drug-drug salt nanocrystals with a particle size of 100-500 nm and a dispersibility index of 0.000-0.300, and then freeze-drying them to obtain vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen drug-drug salt nanocrystal powder.

[0062] As one implementation scheme, the average particle size distribution and morphological characteristics of the vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen drug-drug salt nanocrystals are basically as follows: Figure 8 As shown in 9, 18 and 19.

[0063] Fourthly, the present invention provides a pharmaceutical composition comprising the drug-drug salt nanocrystals and pharmaceutically acceptable excipients.

[0064] Fifthly, the present invention provides the use of the aforementioned drug-drug salt nanocrystals, or drug compositions thereof, in the preparation of antidepressant drugs.

[0065] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0066] 1. The drug-drug salt nanocrystals provided by this invention can achieve synergistic effects of vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen, significantly enhancing antidepressant activity.

[0067] 2. The drug-drug salt nanocrystals provided by this invention utilize their sustained-release effect to extend the dosing interval, thereby achieving long-term treatment and solving the clinical problem of patients with depression needing to take medication frequently.

[0068] 3. The drug-drug salt nanocrystal preparation method provided by the present invention is simple to operate, has low energy consumption, is easy to repeat, has good product reproducibility, and has a high yield, making it suitable for industrial production.

[0069] Terminology Definitions and Explanations

[0070] In the specification and claims of this application, unless otherwise stated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. However, for a better understanding of this disclosure, definitions and explanations of some related terms are provided below. Furthermore, in the event of any discrepancy between the definitions and explanations of terms provided herein and their commonly understood meanings by those skilled in the art, the definitions and explanations provided herein shall prevail.

[0071] The term "about" is used in this application to mean approximately, around, roughly, or approximately. When the term "about" is used in conjunction with a numerical range, the range is modified by expanding the upper and lower limits of the stated numerical range. Unless otherwise stated, the term "about" is used herein to modify the upper and lower limits of the stated value by a numerical value with a 10% deviation.

[0072] The term "pharmaceuticalally acceptable excipient" refers to excipients that do not cause significant irritation to the organism and do not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, etc. Attached Figure Description

[0073] Figure 1 This is a single-crystal structure diagram of vortioxetine-diclofenac drug-drug salt;

[0074] Figure 2 DSC images of vortioxetine-diclofenac drug-drug salt crystals and their drug-drug salt nanocrystals;

[0075] Figure 3 PXRD patterns of vortioxetine-diclofenac drug-drug salt crystals and their drug-drug salt nanocrystals;

[0076] Figure 4 IR chromatogram of vortioxetine-diclofenac drug-drug salt crystals;

[0077] Figure 5 The TG spectrum of vortioxetine-diclofenac drug-drug salt crystals;

[0078] Figure 6 X-ray powder diffraction pattern for long-term stability test of vortioxetine-diclofenac drug-drug salt crystals;

[0079] Figure 7 X-ray powder diffraction pattern of vortioxetine-diclofenac drug-drug salt crystals in accelerated stability test;

[0080] Figure 8 Average particle size distribution of vortioxetine-diclofenac drug-drug salt nanocrystals

[0081] Figure 9 Morphological characteristics of vortioxetine-diclofenac drug-drug salt nanocrystals

[0082] Figure 10 Stability diagrams of vortioxetine-diclofenac drug-drug salt nanocrystals under (a) 4℃ and (b) 25℃ conditions;

[0083] Figure 11 The single-crystal structure diagram of vortioxetine-(S)-flurbiprofen drug-drug salt;

[0084] Figure 12 DSC images of vortioxetine-(S)-flurbiprofen drug-drug salt crystals and their drug-drug salt nanocrystals;

[0085] Figure 13 PXRD patterns of vortioxetine-(S)-flurbiprofen drug-drug salt crystals and their drug-drug salt nanocrystals;

[0086] Figure 14 IR chromatogram of vortioxetine-(S)-flurbiprofen drug-drug salt crystals;

[0087] Figure 15 The TG spectrum of vortioxetine-(S)-flurbiprofen drug-drug salt crystals;

[0088] Figure 16 X-ray powder diffraction pattern for long-term stability test of vortioxetine-(S)-flurbiprofen drug-drug salt crystals;

[0089] Figure 17 X-ray powder diffraction pattern of vortioxetine-(S)-flurbiprofen drug-drug salt crystals in accelerated stability test;

[0090] Figure 18 The average particle size distribution of vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals;

[0091] Figure 19 Morphological characteristics of vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals;

[0092] Figure 20 Stability diagrams of vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals under (a) 4℃ and (b) 25℃ conditions;

[0093] Figure 21 The equilibrium solubility results of vortioxetine and diclofenac or (S)-flurbiprofen drug-drug salt nanocrystals in buffer solutions at pH 1.2, 4.5, 6.8, and 7.4 are presented.

[0094] Figure 22 Dissolution profiles of vortioxetine and diclofenac or (S)-flurbiprofen drug-drug salt nanocrystals in phosphate buffer solution at pH 6.8.

[0095] Figure 23 This diagram illustrates the effects of vortioxetine and diclofenac drug-drug salt nanocrystals on the behavior of LPS-induced depression model mice (Figure a is the experimental protocol flowchart. Figures b and d show the total activity time of mice in the first 5 minutes of the open field experiment on days 6, 11, and 13, respectively. Figures eg and e show the cumulative immobility time of mice in the tail suspension experiment on days 6, 11, and 13, respectively).

[0096] Figure 24 A schematic diagram showing the effect of vortioxetine and diclofenac drug-drug salt nanocrystals on inflammatory factors in the brains of LPS-induced depressed mice (Figure (a) shows the TNF-α content in the hippocampus of mice, and Figure (b) shows the IL-1β content in the hippocampus of mice).

[0097] Figure 25 A schematic diagram showing the effect of vortioxetine and diclofenac drug-drug salt nanocrystals on the neurotransmitter (5-HT) in the brains of LPS-induced depressed mice.

[0098] Figure 26Representative histopathological images of muscle tissue after intramuscular injection of vortioxetine and diclofenac drug-drug salt nanocrystals (black arrows indicate: granulocyte and macrophage infiltration; myofibril degeneration, manifested as loss of striations and fragmentation, scale bar = 100 micrometers). Detailed Implementation

[0099] The invention can be better understood from the following embodiments. However, the descriptions in the embodiments are for illustrative purposes only and should not, and will not, limit the invention as detailed in the claims.

[0100] In the following examples, "PM" refers to a mixture of two single drugs physically mixed in a molar ratio of 1:1, such as... Figure 2 The VOT-DIC-PM group represents the physical mixing of the drugs vortioxetine (VOT) and diclofenac (DIC) in a 1:1 molar ratio. (See attached image.) Figure 12 The VOT-SFLU-PM group represents the physical mixing of the drug vortioxetine (VOT) and the drug (S)-flurbiprofen (SFLU) in a molar ratio of 1:1.

[0101] The instruments used in this invention for detecting the structure and properties of drug-drug salt crystals and their nanocrystals are as follows:

[0102] (1) X-ray powder diffractometer, manufactured by Panaco GmbH, Netherlands, model X'Perrt PRO MPD, Cu-K(α), tube voltage 40kV, tube current 40mA, scanning speed 2° / min, 2θ range 3-60°;

[0103] (2) Differential scanning calorimeter, Netzsch DSC 200F3, Germany. This invention uses a nitrogen atmosphere and a heating rate of 10K / min.

[0104] (3) Fourier transform infrared spectrometer, manufactured by Bruker GmbH, Germany, model Vertex 70, with an absorbance range of 4000–500 cm⁻¹. -1 ;

[0105] (4) Thermogravimetric analyzer, Netzsch TG 209F3, Germany. This invention uses a nitrogen atmosphere. The test temperature range is selected from room temperature to 800℃ (inclusive). The test mode is selected as heating curve. The specific test temperature range is 30~350 (℃). The heating rate is 10℃ / min.

[0106] (5) Single-crystal X-ray diffractometer, detector model: Rigaku HyPix, this invention uses a nitrogen atmosphere, low-temperature detection at 100K, graphite monochromator, copper target X-ray, wavelength of Bruker Apex 2 CCD diffractometer, graphite monochromator, molybdenum target X-ray, wavelength:

[0107] (6) Nanoparticle size potentiometer, Malvern Panaco Ltd., UK, model: Zetasizer Nano ZS90, temperature 25℃, measurement at a 90° detection angle.

[0108] Example 1: Preparation and Characterization of Vortioxetine-Diclofenac Drug-Drug Salt Crystals (VOT-DIC)

[0109] 149.3 mg (0.5 mmol) of vortioxetine was dissolved in 3 mL of n-propanol, and 148.1 mg (0.5 mmol) of diclofenac was dissolved in 4 mL of n-propanol. The two systems were then mixed in a test tube, and 3 mL of purified water was added. The mixture was allowed to stand at 20 ± 5 °C for one day. Crystals precipitated, and the solid was filtered. The obtained solid was washed with n-heptane, dried under vacuum, and collected to give 223.1 mg of the title compound. The product obtained is vortioxetine-diclofenac drug-drug salt crystals.

[0110] The single-crystal X-ray diffraction results of the drug-drug salt crystals obtained in Example 1 are as follows: Figure 1 As shown in the table below, the single-crystal data are as follows:

[0111]

[0112] The differential thermal (DSC) spectrum of the vortioxetine-diclofenac drug-drug salt crystals obtained in Example 1 is shown below. Figure 2 As shown, the melting point of the drug-drug salt crystals is different from that of the active pharmaceutical ingredient and the co-formed drug. There is an endothermic peak at 190.5℃, which proves that a new phase is formed.

[0113] The powder X-ray diffraction (PXRD) pattern of the vortioxetine-diclofenac drug-drug salt crystals obtained in Example 1 is shown below. Figure 3 As shown, characteristic peaks are present at diffraction angles 2θ of 5.680°, 7.903°, 11.218°, 13.519°, 14.383°, 16.781°, 18.523°, 19.278°, 20.182°, 20.883°, 22.142°, 22.381°, 23.508°, 24.702°, 28.041°, and 33.763°, proving the formation of a new crystalline phase. The XRPD diffraction peak parameters are shown in the table below.

[0114] 1 5.680 15.5476 6165 100 3 11.218 7.8808 4814 78.1 6 16.781 5.2789 1644 26.7 12 22.381 3.9691 5664 91.9 15 28.041 3.1796 2691 43.7

[0115] The infrared spectrum (IR) of the vortioxetine-diclofenac drug-drug salt crystals obtained in Example 1 is as follows: Figure 4As shown: Due to intermolecular ionic interactions, the -NH- and C=O groups in vortioxetine, along with the -OH groups formed in the drug, all exhibit red-shift or blue-shift in the IR spectrum of the vortioxetine drug-drug salt crystals, at 3433.62 cm⁻¹. -1 1634.70cm -1 1578.16cm -1 1558.55cm -1 1469.58cm -1 1450.78cm - 1. 1373.43cm -1 1315.38cm -1 1230.18cm -1 1151.23cm -1 1044.82cm -1 813.78cm -1 746.51cm -1 The presence of a characteristic absorption peak further confirms the formation of vortioxetine-diclofenac drug-drug salt crystals.

[0116] The TG curve of the vortioxetine-diclofenac drug-drug salt crystals obtained in Example 1 is shown below. Figure 5 The diagram shows the weight loss starting at 240.9±1℃, corresponding to the melting and decomposition process of vortioxetine-diclofenac drug-drug salt crystals. (From...) Figure 5 Thermogravimetric analysis data of vortioxetine and diclofenac drug-drug salt crystals show that vortioxetine and the co-formed drug do not have solvent weight loss peaks before their respective melting points, indicating that they do not contain solvent. After the temperature reaches the corresponding melting point, the sample begins to decompose gradually.

[0117] Example 2 Preparation and Characterization of Votexetine-Diclofenac Drug-Drug Salt Nanocrystals (VOT-DIC nanocrystaI)

[0118] Weigh 200 mg of vortioxetine-diclofenac drug-drug salt crystals (VOT-DIC, prepared in Example 1), add 10 ml of purified water and 0.15 g of Tween 80, and run in a ball mill at 600 rpm for 10 minutes to obtain a vortioxetine-diclofenac drug-drug salt nanocrystal suspension. Then, freeze-dry the suspension to obtain powdered drug-drug salt nanocrystals.

[0119] The differential thermal (DSC) spectrum of the vortioxetine-diclofenac drug-drug salt nanocrystals obtained in Example 2 is shown below. Figure 2As shown, the nanocrystals exhibit an endothermic peak at 185.3℃. The change in melting point between the nanocrystals and the vortioxetine-diclofenac drug-drug salt crystals can be attributed to the interaction with the co-ground surfactant during the preparation process.

[0120] The powder X-ray diffraction (PXRD) pattern of the vortioxetine-diclofenac drug-drug salt nanocrystals obtained in Example 2 is shown below. Figure 3 As shown, characteristic peaks are observed at diffraction angles 2θ of 5.663°, 7.877°, 11.193°, 13.501°, 14.418°, 16.794°, 18.546°, 19.285°, 20.179°, 20.878°, 22.068°, 22.370°, 23.113°, 25.094°, 28.016°, and 33.371°, which are highly consistent with the drug-drug salt, confirming that the drug-drug salt nanocrystals maintain the same crystal structure. The XRPD diffraction peak parameters are shown in the table below.

[0121] l 5.663 15.5928 3572 100 3 11.193 7.8986 1477 41.4 6 16.794 5.2748 808 22.6 11 22.370 3.9710 1149 32.2 14 28.016 3.1823 477 13.3

[0122] The average particle size distribution and morphological characteristics of the vortioxetine-diclofenac drug-drug salt nanocrystals obtained in Example 2 are shown in the figure below. Figure 8 As shown in Figure 9, the average particle size of the vortioxetine-diclofenac drug-drug salt nanocrystals is 316.57±11.46 μm, and the polydispersity index (PDI) is 0.080±0.072, which confirms that the particles are uniformly distributed and that the particles are mainly spherical.

[0123] Example 3 Preparation and characterization of vortioxetine-(S)-flurbiprofen drug-drug salt crystals (VOT-SFLU)

[0124] 149.23 mg (0.5 mmol) of vortioxetine and 122.1 mg (0.5 mmol) of (S)-flurbiprofen were dissolved in 5 mL of acetone, followed by the addition of 4 mL of n-hexane and allowed to stand at 20 ± 5 °C. After one day, crystals precipitated. The solution was filtered, and the resulting solid was washed with n-heptane, dried under vacuum, and collected to yield 253.84 mg of the title compound. The product obtained is vortioxetine-(S)-flurbiprofen drug-drug salt crystals.

[0125] The single-crystal X-ray diffraction results of the drug-drug salt crystals obtained in Example 3 are as follows: Figure 11 As shown in the table below.

[0126]

[0127] The differential thermal (DSC) spectrum of the vortioxetine-(S)-flurbiprofen drug-drug salt crystals obtained in Example 3 is shown below. Figure 12 As shown: The melting point of the drug-drug salt crystals is different from that of the active pharmaceutical ingredient and the co-formed drug. There is an endothermic peak at 127.0℃, which proves that a new phase is formed.

[0128] The powder X-ray diffraction (PXRD) pattern of the vortioxetine-(S)-flurbiprofen drug-drug salt crystals obtained in Example 3 is shown below. Figure 13 As shown, characteristic peaks are present at diffraction angles 2θ of 5.863°, 9.421°, 11.362°, 13.544°, 15.221°, 16.499°, 18.380°, 19.257°, ​​19.863°, 20.401°, 22.060°, 23.725°, 24.276°, 25.441°, 26.460°, 28.241°, and 31.658°, proving the formation of a new crystalline phase. The XRPD diffraction peak parameters are shown in the table below.

[0129] 2 9.421 9.3795 2867 100 3 11.362 7.7818 1656 57.7 6 16.449 5.3684 1460 50.9 9 19.863 4.4663 2276 79.4 12 23.725 3.7473 2789 97.3

[0130] The infrared spectrum (IR) of the vortioxetine-(S)-flurbiprofen drug-drug salt crystals obtained in Example 3 is as follows: Figure 14 As shown: Due to intermolecular ionic interactions, the -NH- and C=O groups in vortioxetine, along with the -OH groups formed in the drug, all exhibit red-shift or blue-shift in the IR spectrum of the vortioxetine drug-drug salt crystals, at 3434.65 cm⁻¹. -1 1620.78cm -1 1580.05cm -1 1471.23cm -1 1383.22cm -1 1351.34cm -1 1229.42cm -1 1125.53cm -1 1044.85cm -1 926.74cm -1 873.08cm -1 766.84cm -1 696.48cm -1 The presence of a characteristic absorption peak further confirms the formation of vortioxetine-(S)-flurbiprofen drug-drug salt crystals.

[0131] The TG curve of the vortioxetine-(S)-flurbiprofen drug-drug salt crystals obtained in Example 3 is shown below. Figure 15 The diagram shows the weight loss starting at 249.3±1℃, corresponding to the melting and decomposition process of vortioxetine-(S)-flurbiprofen drug-drug salt crystals. (From...) Figure 15Thermogravimetric analysis data of vortioxetine-(S)-flurbiprofen drug-drug salt crystals show that vortioxetine and the co-formed drug do not have solvent weight loss peaks before their respective melting points, indicating that they do not contain solvent. After the temperature reaches the corresponding melting point, the sample begins to decompose gradually.

[0132] Example 4 Preparation and characterization of vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals (VOT-SFLU nanocrystal)

[0133] Weigh 200 mg of vortioxetine-(S)-flurbiprofen drug-drug salt crystals (VOT-SFLU, prepared in Example 3), add 10 ml of purified water and 0.15 g of Tween 80, and run in a ball mill at 600 rpm for 10 minutes to obtain a vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystal suspension. Then, freeze-dry the suspension to obtain powdered drug-drug salt nanocrystals.

[0134] The differential thermal (DSC) spectrum of the vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals obtained in Example 4 is shown below. Figure 12 As shown, the nanocrystals exhibit an endothermic peak at 122.7℃. The change in melting point between the nanocrystals and the vortioxetine-(S)-flurbiprofen drug-drug salt crystals can be attributed to the interaction with the co-milled surfactant during the preparation process.

[0135] The powder X-ray diffraction (PXRD) pattern of the vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals obtained in Example 4 is shown below. Figure 13 As shown, characteristic peaks are observed at diffraction angles 2θ of 5.527°, 9.129°, 11.053°, 13.249°, 15.628°, 16.186°, 18.176°, 19.630°, 20.156°, 22.223°, 23.428°, 23.996°, 25.174°, 26.112°, 28.752°, and 31.442°, which are highly consistent with the drug-drug salt, confirming that the drug-drug salt nanocrystals maintain the same crystal structure. The XRPD diffraction peak parameters are shown in the table below.

[0136] 2 9.129 9.6790 2253 100 3 11.053 7.9987 846 37.5 6 16.186 5.4717 607 26.9 8 19.630 4.5187 905 40.2 12 23.996 3.7100 293 13

[0137] The average particle size distribution and morphological characteristics of the vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals obtained in Example 4 are shown in the figure below. Figure 18 As shown in Figure 19, the average particle size of the vortioxetine-diclofenac drug-drug salt nanocrystals was 250.3±7.46 nm, and the polydispersity index (PDI) was 0.117±0.027, confirming that the particles were uniformly distributed and that the particles were mainly spherical.

[0138] Example 5: Equilibrium Solubility Experiment

[0139] Excess vortioxetine and the obtained drug-drug salt nanocrystals were placed in a 20 mL glass test tube, and 5 mL of pH 1.2 hydrochloric acid solution, pH 4.5 acetate buffer solution, pH 6.8 phosphate buffer solution and pH 7.4 phosphate buffer solution were added respectively. The sample was shaken thoroughly in a constant temperature shaker at 37℃ for 24 hours at a shaking speed of 100 rpm. The supernatant was filtered through a 0.45 μm organic membrane filter, and the filtrate was then analyzed by Shimadzu liquid chromatography. Separation was performed using a C18 column (4.6 × 250 mm, 5.0 μm, Shimadzu) at 35℃. Mobile phase A was 10 mM potassium dihydrogen phosphate (pH 2.5), and mobile phase B was acetonitrile. A gradient elution program was used as follows: time (min) / B% (v / v): 0 / 20; 8 / 80; 15 / 80; 15.01 / 20; 25 / 20, flow rate was 1 mL / min, injection volume was 10 μL, and detection wavelength was 254 nm. The peak area was recorded, and the equilibrium solubility was calculated by substituting it into the standard curve. Figure 21 As shown, in a pH 6.8 phosphate buffer solution, the solubility of vortioxetine is 0.042 mg / ml. In vortioxetine-(S)-flurbiprofenic acid drug-drug salt nanocrystals, the solubility of vortioxetine is 0.0604 mg / ml, an increase of 1.4 times. In contrast, the solubility of vortioxetine in vortioxetine-diclofenac drug-drug salt nanocrystals is 0.011 mg / ml, a decrease of 0.38 times. The low solubility of vortioxetine-diclofenac drug-drug salt nanocrystals is beneficial for achieving sustained drug release, thereby achieving long-term therapeutic effects.

[0140] Example 6 Powder dissolution experiment

[0141] To investigate the dissolution of nanocrystals, evaluate the release and absorption characteristics of the drug in vivo, and ensure its safety and efficacy, this invention conducted an in vitro powder dissolution test on vortioxetine and its drug-drug salt nanocrystals. The powder dissolution rate was determined according to the paddle method (FADT-1202, Shanghai Fukesi) required by the Chinese Pharmacopoeia. The specific test method is as follows: First, 100 mg of vortioxetine and an equal amount of drug-drug salt nanocrystal powder containing vortioxetine were accurately weighed and poured into 500 ml of pH 6.8 phosphate buffer solution. The test temperature was 37 ± 0.5 °C, and the paddle rotation speed was 50 rpm. Sample solutions of 2 mL were collected at time points of 5, 10, 15, 30, 45, 60, 90, 120, 180, 240, 300, 360, 480, 720, 1080, and 1440 min. Immediately after collection, an equal volume of buffer solution was added to the original solution to make up the volume to avoid errors. The extracted sample solution was analyzed by HPLC (Shimadzu). The separation was performed using a C18 column (4.6×250mm, 5.0μm, Shimadzu), with a column temperature of 35℃. Mobile phase A was 10mM potassium dihydrogen phosphate (pH 2.5), and mobile phase B was acetonitrile. The gradient elution program was as follows: time (min) / B% (v / v): 0 / 20; 8 / 80; 15 / 80; 15.01 / 20; 25 / 20, flow rate was 1mL / min, injection volume was 10μL, and detection wavelength was 254nm.

[0142] Figure 22 The figure shows the dissolution curves of the obtained vortioxetine and its drug-drug salt nanocrystals in a phosphate buffer solution at pH 6.8. As can be seen from the figure, the dissolution rate (70%) of the vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals provided by this invention is comparable to that of pure vortioxetine (67%). However, the equilibrium solubility of the vortioxetine-diclofenac drug-drug salt nanocrystals is significantly lower, approximately 9%. The release curve of the vortioxetine-diclofenac drug-drug salt nanocrystals is consistently lower than that of the parent drug VOT, indicating that it possesses sustained-release dissolution characteristics.

[0143] Example 7 Stability Experiment

[0144] Long-term stability tests were conducted on the obtained drug-drug salt crystals of vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen using a constant temperature and humidity chamber. The crystals were stored for 8 weeks at 25±2℃ and 60±5% relative humidity, and samples were taken at 0 days, 2 weeks, 4 weeks, and 8 weeks for X-ray powder diffraction analysis. (See relevant spectra...) Figure 6 and Figure 16 As shown in the figure, the X-ray powder diffraction pattern remains unchanged, indicating that the crystal structure has not changed and its thermal stability is good.

[0145] Accelerated stability tests were conducted on the obtained drug-drug salt crystals of vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen using a constant temperature and humidity chamber. The crystals were stored for 8 weeks at 40±2℃ and 75±5% relative humidity, and samples were taken at 0, 2, 4, and 8 weeks for X-ray powder diffraction analysis. (See relevant spectra...) Figure 7 and Figure 17 As shown in the figure, the X-ray powder diffraction pattern remains unchanged, indicating that the crystal structure has not changed and its wet stability is good.

[0146] Vortioxetine nanocrystals with diclofenac or (S)-flurbiprofen were stored at 4°C and 25°C for 21 days. Samples were collected on days 1, 3, 5, 7, 14, and 21, and particle size and particle size distribution index were analyzed using a particle size analyzer. (See related chromatograms.) Figure 10 and Figure 20 As shown in the figure, the particle size and particle diameter distribution are good and there are no significant changes, indicating that the nanocrystalline system is stable.

[0147] Example 8: Evaluation of the antidepressant activity of vortioxetine-diclofenac drug-drug salt nanocrystals in an LPS-induced depression model.

[0148] Lipopolysaccharide (LPS) is commonly used in depression models to rapidly induce depression and provides experimental evidence for the screening and efficacy evaluation of antidepressants. This model is considered to be able to effectively elicit depression-like symptoms, and the behavioral characteristics of animals will change after stress. Moreover, the model is stable and has high reference value as a depression model.

[0149] To investigate the effects of vortioxetine and diclofenac drug-drug salt nanocrystals on lipopolysaccharide-induced depressive-like behavior, and to evaluate these effects using open field and tail suspension tests, a mouse depressive-like behavior model was established by intraperitoneal injection of LPS (0.83 mg / kg) for 5 consecutive days. The control group received an equal volume of saline. Mice were divided into six groups of eight each: control group, model group (LPS), vortioxetine group (VOT), diclofenac group (DIC), vortioxetine + diclofenac combined administration group (VOT+DIC), and vortioxetine-diclofenac drug-drug salt nanocrystal group (VOT-DIC). Except for the VOT-DIC drug-drug salt nanocrystals, which were administered intramuscularly, all other drugs were dissolved in sterile saline and administered via intraperitoneal injection. All interventions were initiated 24 hours after the first LPS injection.

[0150] Behavioral assessments were performed on days 1, 5, and 7 after initial drug treatment. Figure 23As shown, 5 days after intraperitoneal injection of lipopolysaccharide (LPS), mice exhibited significant depressive-like behaviors, specifically shortened movement time in the open field test and increased immobility time in the tail suspension test; both of these behaviors are core characteristics of depressive-like behaviors. Notably, treatment with VOT-DIC drug-drug salt nanocrystals effectively reversed these behavioral trends. Figure 23 d[F(5,30)=10.46,p<0.0001] represents the time mice spent moving in the open field in the first 5 minutes on the seventh day after treatment. The control group (Control) was 219.7±19.54s, the model group (LPS) was 149.7±7.53s, the vortioxetine group (VOT) was 154.9±29.43s, the diclofenac fentanyl (DIC) group was 158.1±26.46s, the vortioxetine + diclofenac (VOT+DIC) group was 153.3±22.74s, and the vortioxetine-diclofenac (VOT-DIC) group was 191.5±14.99s. From the behavioral data, LPS can significantly reduce the mice's movement and exploration ability in the open field, mainly manifested as a decrease in movement time. After treatment with VOT-DIC drug-drug salt nanocrystals, mice regained their motor and exploratory abilities. Simultaneously, they exhibited a significant antidepressant effect in the tail suspension test, manifested as a significant reduction in immobility time, such as... Figure 23 g[F(5, 30) = 8.497, p < 0.0001] represents the seventh day after treatment. The immobility time in the control group (Control) was 121.7 ± 16.85 s in the last 4 minutes, while it was 178.38 ± 24.91 s in the model group (LPS), 178.6 ± 27.11 s in the vortioxetine group (VOT), 188.54 ± 17.92 s in the diclofenac sodium (DIC) group, and 188.54 ± 17.92 s in the vortioxetine + diclofenac sodium (VOT) group. The immobility time in the VOT+DIC group was 179.8±23.23s, and in the vortioxetine-diclofenac (VOT-DIC) group it was 159.1±6.42s. The data show that the immobility time in the model group (LPS) was significantly higher than that in the control group (Control), indicating a decreased will to live in LPS mice, exhibiting behavioral despair. The VOT-DIC drug-drug salt nanocrystal group significantly reduced immobility time, alleviated despair behavior, and enhanced the will to live in the mice. In conclusion, the improvement effect of vortioxetine-diclofenac drug-drug salt nanocrystals (VOT-DIC) is significantly better than that of vortioxetine or diclofenac monotherapy or their combination therapy, and this effect can last until day 7 after administration.

[0151] Example 9: Effect of vortioxetine-diclofenac drug-drug salt nanocrystals on the content of inflammatory factors in the hippocampus of LPS-induced depressed mice.

[0152] Studies have shown a correlation between the overexpression of inflammatory factors and depression, and related literature has reported the generation of large amounts of inflammatory factors in patients with depression. To investigate whether the antidepressant effect of VOT-DIC drug-drug salt nanocrystals is achieved through anti-neuroinflammatory effects, this invention quantitatively detected the concentrations of pro-inflammatory cytokines IL-1β and TNF-α in the hippocampus after administration of lipopolysaccharide (LPS). The results are as follows... Figure 24 The TNF-α levels shown in a[F(5,6)=2635, p<0.0001] were as follows: Control group 119.7±4.89pg / ml, Model group (LPS) 473.1±6.67pg / ml, Vortioxetine group (VOT) 30.6±3.49pg / ml, Diclofenac group (DIC) 105.1±4.36pg / ml, Vortioxetine + Diclofenac (VOT+DIC) group 150.58±4.82pg / ml, and Vortioxetine-Diclofenac (VOT-DIC) group 25.5±1.43pg / ml. Figure 24 As shown in b[F(5,6)=6260, p<0.0001], the IL-1β levels in each group were as follows: Control group 69.4±0.62pg / ml, Model group (LPS) 275.5±3.37pg / ml, Vortioxetine group (VOT) 15.8±0.45pg / ml, Diclofenac group (DIC) 60.6±0.59pg / ml, Vortioxetine + Diclofenac (VOT+DIC) group 83±2.36pg / ml, and Vortioxetine - Diclofenac (VOT-DIC) group 13.5±0.45pg / ml. LPS induced a significant increase in TNF-α and IL-1β levels in the hippocampus of mice, successfully establishing an inflammation model. After treatment with VOT-DIC drug-drug salt nanocrystals, the expression of pro-inflammatory factors was continuously suppressed, and this inhibitory effect remained significant on day 7 post-treatment.

[0153] Example 10: Effect of vortioxetine-diclofenac drug-drug salt nanocrystals on neurotransmitter levels in the brains of LPS-induced depressed mice

[0154] Inflammation leads to the overexpression of NF-κB and COX-2. Studies have reported that inflammation affects neurotransmitter levels in the brain, resulting in reduced 5-HT and NE levels. Furthermore, neurotransmitters are associated with depressive-like behaviors. To assess whether VOT-DIC drug-drug salt nanocrystals can increase neurotransmitter levels, this invention detected 5-HT levels in the hippocampus of mice. Figure 25[F(5,6)=126.5, p<0.0001], the 5-HT content in each group was as follows: Control group 11.9±0.2 ng / ml, Model group (LPS) 7.7±0.25 ng / ml, Vortioxetine group (VOT) 9.4±0.01 ng / ml, Diclofenac group (SFLU) 8.1±0.13 ng / ml, Vortioxetine + Diclofenac group (VOT+DIC) 9.2±0.28 ng / ml, Vortioxetine-Diclofenac group (VOT-DIC) 10.8±0.3 ng / ml. The results showed that LPS treatment significantly reduced 5-HT levels, while the intervention of VOT-DIC nanocrystals reversed this downward trend and caused an increase in 5-HT concentration. Figure 25 Furthermore, in an LPS-induced depression model, VOT-DIC drug-drug salt nanocrystals showed superior serotonin (5-HT) enhancement in the hippocampus compared to single drugs or their combinations. These results suggest that VOT-DIC drug-drug salt nanocrystals exert a sustained antidepressant effect in LPS-treated mice, likely by maintaining elevated 5-HT levels in the hippocampus.

[0155] Example 11: Muscle Stimulation Assessment of Vortioxetine-Diclofenac Drug-Drug Salt Nanocrystals

[0156] To assess potential muscle irritation, cross-sectional tissue samples were stained and histopathologically analyzed using an optical microscope. Figure 26 At all observation time points (days 0, 1, 3, 5, and 7 post-injection), the saline control group showed no signs of inflammatory cell infiltration or myofiber damage, confirming its biocompatibility. Conversely, histological sections of the 1.7% acetic acid group showed severe pathological changes, including loss of myofiber striations, extensive necrosis, interstitial edema, and dense granulocytic / macrophage infiltration. Notably, this damage persisted and was not fully resolved by day 7. For vortioxetine-diclofenac drug-drug salt nanocrystals (19.92 mg / kg), intramuscular injection induced transient and mild local damage and inflammation on days 1 and 3, which resolved completely within 5 days. In summary, histological examination indicates that VOT-DIC drug-drug salt nanocrystals induced a reversible inflammatory response.

[0157] The vortioxetine-diclofenac or vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals provided by this invention can be used to prepare drugs for the prevention and / or treatment of depression, and have broad application prospects.

[0158] In summary, this invention selects a suitable nonsteroidal anti-inflammatory drug (NSAID) and vortioxetine to form drug-drug salt crystals, which are then further prepared into nanocrystals. The resulting drug-drug salt nanocrystals can achieve synergistic effects between vortioxetine and diclofenac or between vortioxetine and (S)-flurbiprofen, enhancing antidepressant activity while achieving long-acting sustained release and comprehensively improving therapeutic efficacy.

[0159] The embodiments of the present invention have been described in detail above with reference to specific examples. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention.

Claims

1. A drug-drug salt nanocrystal, characterized in that: The nanocrystals contain a) a drug-drug salt of vortioxetine and diclofenac or a drug-drug salt of vortioxetine and (S)-flurbiprofen; and b) a stabilizer.

2. The drug-drug salt nanocrystals according to claim 1, characterized in that, The drug-drug salt nanocrystals mentioned are vortioxetine-diclofenac drug-drug salt nanocrystals; Alternatively, the drug-drug salt nanocrystals are vortioxetine-diclofenac drug-drug salt nanocrystals, and the powder X-ray diffraction pattern of the nanocrystals includes the following characteristic peaks shown by the 2θ angle: 5.656°, 7.859°, 11.179°, 13.481°, 14.381°, 16.781°, 18.521°, 19.240°, 20.178°, 20.860°, 22.359°, 23.080°, 25.077°, 28.883°, 33.400°, with a 2θ value accuracy of ±0.2°; Alternatively, the nanocrystals are drug-drug salt nanocrystals of vortioxetine-diclofenac, and the differential scanning calorimetry curve of the nanocrystals has an endothermic peak at 185.3±1℃. Alternatively, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, wherein the nanocrystal particle size is 100-500 nm and the dispersibility index is 0.000-0.

300.

3. The drug-drug salt nanocrystals according to claims 1-2, characterized in that, The nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, which are prepared from vortioxetine-diclofenac drug-drug salt as raw materials, wherein the molar ratio of vortioxetine and diclofenac is approximately 1:

1. Alternatively, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, prepared from vortioxetine-diclofenac drug-drug salt as a raw material, wherein the vortioxetine-diclofenac drug-drug salt crystal is monoclinic with space group P21 / C and cell parameters as follows: α = 90°, β = 102.073(3)°, γ = 90°, and the unit cell volume is Alternatively, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, prepared from vortioxetine-diclofenac drug-drug salt as raw material. The powder X-ray diffraction pattern of the vortioxetine-diclofenac drug-drug salt crystals includes the following characteristic peaks at 2θ angles: 5.663°, 7.877°, 11.193°, 13.501°, 14.418°, 16.794°, 18.546°, 19.285°, 20.179°, 20.878°, 22.068°, 22.370°, 23.113°, 25.094°, 28.016°, and 33.371°, with an accuracy of ±0.2° for the 2θ values. Alternatively, the nanocrystals may be drug-drug salt nanocrystals of vortioxetine and diclofenac, prepared from vortioxetine-diclofenac drug-drug salt as a raw material, wherein the infrared absorption spectrum of the vortioxetine-diclofenac drug-drug salt crystals is at 3433.62 cm⁻¹. -1 1634.70cm -1 1578.16cm -1 1558.55cm -1 1469.58cm -1 1450.78cm -1 1373.43cm -1 1315.38cm -1 1230.18cm -1 1151.23cm -1 1044.82cm -1 813.78cm -1 746.51cm -1 There is an absorption peak at this point; Alternatively, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and diclofenac, prepared from vortioxetine-diclofenac drug-drug salt as raw material, wherein the differential scanning calorimetry curve of the vortioxetine-diclofenac drug-drug salt crystals has an endothermic peak at 190.5±1℃, and the thermogravimetric analysis curve shows weight loss starting from 240.9±1℃.

4. The drug-drug salt nanocrystals according to claim 1, characterized in that, The drug-drug salt nanocrystals mentioned are vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals; Alternatively, the drug-drug salt nanocrystals are vortioxetine-(S)-flurbiprofen drug-drug salt nanocrystals, and the powder X-ray diffraction pattern of the nanocrystals includes the following characteristic peaks shown by the 2θ angle: 5.600°, 9.159°, 11.100°, 13.276°, 15.624°, 16.202°, ​​18.200°, 19.641°, 20.120°, 22.280°, 23.476°, 23.999°, 25.201°, 26.201°, 28.619°, 31.740°, with a 2θ value accuracy of ±0.2°; Alternatively, the nanocrystals are drug-drug salt nanocrystals of vortioxetine-(S)-flurbiprofen, and the differential scanning calorimetry curve of the nanocrystals shows endothermic activity at 122.7±1℃. Alternatively, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, wherein the nanocrystal particle size is 100-500 nm and the dispersibility index is 0.000-0.

300.

5. The drug-drug salt nanocrystals according to claims 1 and 4, characterized in that, The nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, and the nanocrystals are prepared using vortioxetine-(S)-flurbiprofen drug-drug salt as raw material, wherein the molar ratio of vortioxetine and (S)-flurbiprofen is approximately 1:

1. Alternatively, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, prepared from vortioxetine-(S)-flurbiprofen drug-drug salt, wherein the vortioxetine-(S)-flurbiprofen drug-drug salt crystal is monoclinic with space group P212121 and cell parameters as follows: α = 90°, β = 90°, γ = 90°, cell volume is Alternatively, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, prepared from vortioxetine-(S)-flurbiprofen drug-drug salt. The powder X-ray diffraction pattern of the vortioxetine-(S)-flurbiprofen drug-drug salt crystals includes the following characteristic peaks at 2θ angles: 5.527°, 9.129°, 11.053°, 13.249°, 15.628°, 16.186°, 18.176°, 19.630°, 20.156°, 22.223°, 23.428°, 23.996°, 25.174°, 26.112°, 28.752°, and 31.442°, with an accuracy of ±0.2° for the 2θ values. Alternatively, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, prepared from vortioxetine-(S)-flurbiprofen drug-drug salt, wherein the infrared absorption spectrum of the vortioxetine-(S)-flurbiprofen drug-drug salt crystals is at 3434.65 cm⁻¹. -1 1620.78cm -1 1580.05cm -1 1471.23cm -1 1383.22cm -1 1351.34cm -1 1229.42cm -1 1125.53cm -1 1044.85cm -1 926.74cm -1 873.08cm -1 766.84cm -1 696.48cm -1 There is an absorption peak at this point; Alternatively, the nanocrystals are drug-drug salt nanocrystals of vortioxetine and (S)-flurbiprofen, which are prepared from vortioxetine-(S)-flurbiprofen drug-drug salt as raw material. The differential scanning calorimetry curve of the vortioxetine-(S)-flurbiprofen drug-drug salt crystals has an endothermic peak at 127.0±1℃, and the thermogravimetric analysis curve shows weight loss starting from 249.3±1℃.

6. The drug-drug salt nanocrystals according to any one of claims 1-5, characterized in that: The stabilizer is selected from any one or more of hydroxypropyl methylcellulose, povidone, poloxamer, sodium carboxymethyl cellulose, Tween 80, and Tween 20. The weight ratio of the nanocrystals to the stabilizer is (1-3):(0.5-2).

7. The drug-drug salt nanocrystals according to claim 4, wherein the stabilizer is Tween 80, and the weight ratio of the nanocrystals to the stabilizer is (1-3):(0.5-2).

8. The method for preparing drug-drug salt nanocrystals according to claims 1-7, comprising the following steps: A drug-drug salt of vortioxetine and diclofenac or a drug-drug salt of vortioxetine and (S)-flurbiprofen, Tween 80, and water are mixed, wherein the mass ratio of the drug-drug salt to Tween 80 is (1-3):(0.5-2). The resulting mixture is wet-milled in a ball mill at 600 rpm for 5-20 minutes to obtain vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen drug-drug salt nanocrystals with a particle size of 100-500 nm and a dispersibility index of 0.000-0.

300. The nanocrystals are then freeze-dried to obtain vortioxetine and diclofenac or vortioxetine and (S)-flurbiprofen nanocrystal powder.

9. A pharmaceutical composition comprising the drug-drug salt nanocrystals of claims 1-7 and pharmaceutically acceptable excipients.

10. The use of the drug-drug salt nanocrystals according to any one of claims 1-7, and the composition according to claim 9, in the preparation of antidepressant drugs.

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