Pharmaceutical composition and application thereof

Abstract

The invention relates to a pharmaceutical composition. The pharmaceutical composition comprises two active ingredients: (1) votioxetine or a pharmaceutically acceptable salt thereof; and (2) olanzapine, the weight ratio of the vothioxetine or the pharmaceutically acceptable salt thereof to the olanzapine is (6-50): (3-6), preferably (9-50): 6, and the weight of the two active ingredients is calculated in a free form. The invention further relates to a pharmaceutical preparation prepared from the composition and application of the pharmaceutical composition and the pharmaceutical preparation in prevention and / or treatment of depression and other diseases, and the application comprises the step of applying an effective amount of the pharmaceutical composition or the pharmaceutical preparation to a patient in need. The pharmaceutical composition disclosed by the invention can be used for treating bipolar affective disorder, refractory depression and anxiety and improving cognition, and also has the advantages of small side effect and good patient compliance.

Description

Technical Field

[0001] The present invention belongs to the field of medicine and relates to a pharmaceutical composition and use thereof, and in particular to a pharmaceutical composition and preparation of vortioxetine or a pharmaceutically acceptable salt thereof and olanzapine, and use thereof. Background Art

[0002] Depressive disorder (DDD) is a common psychiatric disorder characterized by depressed mood, a lack of interest, attention deficits, feelings of guilt, and suicidal tendencies, often accompanied by sleep disturbances, behavioral and social dysfunction. It is characterized by high prevalence, relapse rates, and disability. The etiology and pathogenesis of depression remain incompletely understood. In 1965, Schildkraut proposed the classic "monoamine hypothesis," which states that depression is closely linked to low brain levels of monoamines such as serotonin (5-HT), norepinephrine (NE), or dopamine (DA). 5-HT deficiency increases appetite and anxiety, which may explain the bulimia, mood reactivity, and sensitivity to interpersonal rejection seen in atypical depression. NE deficiency reduces energy, interest, and motivation. DA deficiency reduces drive, and these two deficiencies may explain the excessive sleepiness and lethargy seen in atypical depression.

[0003] Bipolar disorder (BD) is a special type of depression. Its full name is bipolar affective disorder, also known as manic-depressive illness. It is characterized by debilitating mood instability with alternating depression and excitement. Patients experience alternating depression and mania. Simply taking antidepressants (such as fluoxetine) will aggravate the patient's manic symptoms and even cause suicidal tendencies.

[0004] Anxiety is an emotional response to potential future threats or dangers. Moderate anxiety is a normal emotional response in humans and animals, motivating them to achieve enhanced physical and intellectual performance. However, excessive anxiety, in terms of intensity and duration, can trigger negative emotional, physical, behavioral, and cognitive symptoms. The lifetime prevalence of anxiety-related disorders in the general Western population is approximately 20-30%, making it the most common neuropsychiatric disorder, with a significantly higher incidence in women than in men (Bandelow, B. & Michaelis, S. Epidemiology of anxiety disorders in the 21st century. Dialogues in Clinical Neuroscience 17, 327-335, 2015). Medical genetics studies have identified numerous anxiety susceptibility genes involved in specific anxiety-related phenotypes and / or the etiology of pathological anxiety, including genes for the serotonin transporter (5-HTT), serotonin 1A receptor (5-HT1AR), monoamine oxidase A, and brain-derived neurotrophic factor (BDNF). Patients with anxiety disorders may have insufficient release of serotonin and noradrenergic neurotransmitters (Connor, KM &

[0005] Davidson, JRT Generalized anxiety disorder: neurobiological and pharmacotherapeutic perspectives. Biological Psychiatry 44, 1286-1294, 1998.), anxiety disorders and major depressive disorder may have a common genetic basis (Tyrer, P. & Baldwin, D. Generalized anxiety disorder. 368, 2006.).

[0006] Cognitive disorders (CDs), also known as neurocognitive disorders (NCDs), are mental health disorders that primarily affect cognitive abilities, including learning, memory, perception, and problem-solving. Acquired neurocognitive disorders include those caused by diseases like Alzheimer's disease (AD) and Parkinson's disease (PD); congenital neurocognitive disorders are more common in developmental delays, such as those on the autism spectrum. Additionally, anxiety disorders, mood disorders, and psychiatric disorders can also affect cognition and memory. Many cognitive impairments are associated with neurotransmitter abnormalities. For example, dopaminergic neuron damage plays a key role in the pathogenesis of Parkinson's disease (PD). Various strategies have been developed to improve dopaminergic neuronal function, including drug supplementation of its precursor L-dopamine, cell transplantation to replace dopaminergic neurons, gene therapy to implant enzyme genes that promote dopamine synthesis to promote dopamine production in the striatum, and neurotrophic factor genes to prevent dopaminergic neuron death or stimulate regeneration and functional recovery of the damaged nigrostriatal system. Therefore, dopamine supplementation can improve cognitive impairment to a certain extent.

[0007] In summary, the occurrence of mental illnesses such as depression, anxiety, and cognitive impairment is related to insufficient levels of relevant neurotransmitters in the brain.

[0008] Treatment-resistant depression (TRD) refers to depression that responds poorly or ineffectively to adequate doses and durations of two or more different types of antidepressants. In clinical practice, physicians often try different treatment combinations based on their experience, but the results are often poor and unpredictable. Few medications are currently marketed for the treatment of TRD. Only Symbyax is approved for this indication in China, the US, and Europe. In December 2003, the FDA approved Eli Lilly's fluoxetine and olanzapine combination capsules, marketed as Symbyax, for the treatment of acute depressive episodes and refractory depression in bipolar I disorder. However, Symbyax is associated with significant gastrointestinal side effects, such as retching and sexual dysfunction, which can affect patient compliance and treatment efficacy. Furthermore, Symbyax is ineffective against the manic symptoms and cognitive impairment associated with bipolar disorder.

[0009] Vortioxetine, also translated into Chinese as Vortioxetine, has a structural formula shown in Formula 1. It is a multi-modal multi-target drug approved by the FDA in September 2013 (six pharmacological targets, namely SERT inhibition, 5-HT 1A Receptor agonist, 5-HT 1B Partial agonist of 5-HT receptors 1D Receptor antagonism 、 Antidepressants (5-HT3 receptor antagonists and 5-HT7 receptor antagonists) primarily affect the 5-HT neurotransmitter system and may also affect neurotransmitter systems such as dopamine and norepinephrine. However, the precise contribution of individual targets to the observed pharmacodynamic characteristics remains unclear, and the specific pharmacological mechanisms are not well defined. Vortioxetine is marketed as a hydrobromide, lactate, and hemihydrobromide salts, and has significant therapeutic effects on depression.

[0010]

[0011] The literature "Efficacy of vortioxetine monotherapy compared with combined therapy vortioxetine and olanzapine in the treatment of major depression-first results", Eur.Pharm.J., 64(1):13-16, Oppa M. et al. reported that there was no statistical difference in MADRS and HAM-A scores between the two groups in the treatment of depression, indicating that the combination therapy and vortioxetine alone were equally effective in treating depression, that is, the combination of vortioxetine and olanzapine had no therapeutic advantage over vortioxetine alone in the treatment of depression.

[0012] Zhu Qian et al., "Observation on the Clinical Efficacy of Olanzapine Combined with Vortioxetine in the Treatment of Depression," Chinese Journal of Medical Science, 2019, vol. 21, pp. 56-59, and Jiao Genlong et al., "Exploration of the Clinical Efficacy of Olanzapine Combined with Vortioxetine in the Treatment of Depression," Clinical Medicine, 2021, vol. 8, pp. 0022-0023, compared the efficacy of vortioxetine and olanzapine combined with olanzapine in the treatment of depression, but did not compare it with vortioxetine alone. Olanzapine is not approved for depression, so these two study groups are not helpful for studies of depression.

[0013] Combination medication affects patients' medication compliance, especially for patients with depression. Medication compliance is very important in the treatment process. By making a compound prescription, the convenience and compliance of medication will be greatly improved.

[0014] In order to solve the above problems, it is necessary to develop a pharmaceutical composition that has a rapid onset of action, few side effects, and good patient compliance, and can treat bipolar disorder, refractory depression, anxiety, and improve cognitive impairment. Summary of the Invention

[0015] The present invention aims to provide a new pharmaceutical composition and a new pharmaceutical preparation, as well as the use of the new pharmaceutical composition and pharmaceutical preparation in the preparation of drugs for treating bipolar disorder, refractory depression, anxiety and improving cognitive impairment.

[0016] While researching antidepressants, the applicant unexpectedly discovered that vortioxetine or its pharmaceutically acceptable salts, particularly its hydrobromide or hemihydrobromide, and olanzapine can be used in combination, significantly outperforming the marketed antidepressant Symbyax in terms of gastrointestinal side effects. Compared to vortioxetine hemihydrobromide alone, the combination of vortioxetine hemihydrobromide and olanzapine significantly increased brain concentrations of 5-HT, dopamine, epinephrine, and norepinephrine, demonstrating a synergistic or potentiating effect between vortioxetine or its pharmaceutically acceptable salts and olanzapine, resulting in improved and faster efficacy in treating conditions such as depression, anxiety, and cognitive impairment. For convenience, vortioxetine hemihydrobromide is referred to herein as "SF001," olanzapine as "Y004," and their combination as "SF001+Y004."

[0017] SF001+Y004 significantly outperforms the marketed antidepressant Symbyax in terms of gastrointestinal side effects. Symbyax can also cause sexual dysfunction, withdrawal effects after discontinuation, and cognitive impairment. The SF001+Y004 combination does not exhibit the adverse side effects of Symbyax. In animal studies, behavior quickly returned to normal after discontinuation of the drug, with no withdrawal effects. In reproductive studies, no sexual dysfunction was observed with the SF001+Y004 combination.

[0018] Compared with SF001, SF001+Y004 significantly increases the concentrations of neurotransmitters such as 5-HT, dopamine, epinephrine, and norepinephrine in the brain through unexpected synergistic or potentiating effects. For example, by increasing dopamine levels, it overcomes the deficiency of SF001 alone in insufficiently improving the lack of satisfaction. SF001+Y004 also improves nerve cell activity and synaptic plasticity, suggesting its potential as a drug for improving cognitive impairment. Overall, increased levels of these neurotransmitters are beneficial for treating depression, anxiety, or improving cognitive impairment.

[0019] The present invention is achieved through the following technical solutions:

[0020] In a first aspect, the present invention provides a pharmaceutical composition comprising (1) vortioxetine or a pharmaceutically acceptable salt thereof; and (2) olanzapine; wherein the weight ratio of vortioxetine or a pharmaceutically acceptable salt thereof to olanzapine is 6-50:3-6, the weights of the two active ingredients being calculated in free form.

[0021] In one or more embodiments, the weight ratio of vortioxetine or a pharmaceutically acceptable salt thereof to olanzapine is 9 to 50:6, for example, 9:6, 10:6, 12:6, 15:6, 18:6, 20:6, 22:6, 25:6, 27:6, 30:6, 35:6, 40:6, 45:6, or 50:6.

[0022] In one or more embodiments, the pharmaceutically acceptable salt is a salt of vortioxetine formed with hydrobromic acid, hydrochloric acid, methanesulfonic acid, fumaric acid, maleic acid, tartaric acid, sulfuric acid, phosphoric acid or nitric acid.

[0023] In one or more embodiments, the pharmaceutically acceptable salt of vortioxetine is vortioxetine hydrobromide or vortioxetine hemihydrobromide.

[0024] In one or more embodiments, the pharmaceutically acceptable salt of vortioxetine is vortioxetine hemihydrobromide.

[0025] In one or more embodiments, the combination of two active ingredients is vortioxetine hemihydrobromide and olanzapine, vortioxetine hydrobromide and olanzapine, preferably a combination of vortioxetine hemihydrobromide and olanzapine.

[0026] In one or more embodiments, the two active ingredients are administered in combination, and the combined administration is simultaneous administration.

[0027] In one or more embodiments, the two active ingredients are administered in combination, and the combined administration is sequential administration.

[0028] In a second aspect, the present invention provides a pharmaceutical preparation, which is composed of a therapeutically effective amount of the pharmaceutical composition described in the first aspect and a pharmaceutically acceptable carrier.

[0029] In one or more embodiments, the pharmaceutical preparation is a tablet, capsule, powder, granule, emulsion, suspension or injection, preferably a tablet.

[0030] In one or more embodiments, the weight ratio of vortioxetine or a pharmaceutically acceptable salt thereof to olanzapine is 5 mg:1.5 mg, 9 mg:6 mg, 9 mg:3 mg, 10 mg:3 mg, 12 mg:6 mg, 15 mg:6 mg, 18 mg:6 mg, 20 mg:6 mg, 22 mg:6 mg, 25 mg:6 mg, 27 mg:6 mg, 30 mg:6 mg, 35 mg:6 mg, 40 mg:6 mg, 45 mg:6 mg or 50 mg:6 mg.

[0031] In one or more embodiments, the weight ratio of vortioxetine or a pharmaceutically acceptable salt thereof to olanzapine is 9 mg:3 mg, 10 mg:3 mg, 8 mg:6 mg or 20 mg:6 mg.

[0032] In one or more embodiments, the pharmaceutical preparation contains one or more fillers, one or more binders, one or more disintegrants, and one or more lubricants, wherein the filler is selected from at least one of mannitol, microcrystalline cellulose, lactose, starch, and sorbitol; the binder is selected from at least one of hydroxypropyl cellulose, ethyl cellulose, and copovidone; the disintegrant is selected from at least one of cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium starch glycolate, and low-substituted hydroxypropyl cellulose; and the lubricant is selected from at least one of talc, magnesium stearate, and sodium stearyl fumarate.

[0033] In one or more embodiments, the pharmaceutical formulation comprises:

[0034]

[0035] In one or more embodiments, the pharmaceutical formulation comprises:

[0036]

[0037] In one or more embodiments, the pharmaceutical formulation comprises:

[0038]

[0039]

[0040] In a third aspect, the present invention provides use of the pharmaceutical composition of the first aspect or the pharmaceutical formulation of the second aspect for preparing a medicament for treating a disease, wherein the disease is affective disorder, depression, anxiety, stress urinary incontinence, vomiting, IBS (irritable bowel syndrome), eating disorders, chronic pain, Alzheimer's disease, ADHD (attention deficit hyperactivity disorder), PTSD (post-traumatic stress disorder), hot flashes, sleep apnea, alcoholism, nicotine or carbohydrate addiction, drug abuse, alcohol or drug abuse, major depressive disorder, postpartum depression, depression associated with bipolar disorder, psychosis, cancer, aging or Parkinson's disease, generalized anxiety disorder, social anxiety disorder, obsessive-compulsive disorder, panic disorder, panic attack, phobia, social phobia, agoraphobia, partial response, resistant depression, cognitive impairment or dysthymia, preferably affective disorder, depression, anxiety, resistant depression or cognitive impairment.

[0041] The pharmaceutical composition of the present invention can be prepared into specific dosage forms and administered by appropriate routes, such as oral, topical (buccal and sublingual), intraperitoneal, intravenous, and intramuscular routes, with oral administration being preferred. Pharmaceutical preparations for oral administration include tablets, capsules, powders, emulsions, and suspensions, while pharmaceutical preparations for intravenous administration include injections.

[0042] The pharmaceutical compositions or formulations of the present invention can be prepared using methods known to those skilled in the art. Pharmaceutically acceptable carriers include pharmaceutical carriers, diluents, excipients, or other additives commonly used in the art. Pharmaceutical carriers include inert solid diluents such as lactose, sucrose, cyclodextrin, talc, agar, pectin, gum arabic, stearic acid, lower alkyl ethers of cellulose, corn starch, potato starch, magnesium stearate, gelatin, and sterile aqueous solutions. The pharmaceutical compositions or formulations of the present invention may also incorporate other excipients or additives commonly used for coloring, flavoring, preservatives, and the like, as long as they are compatible with the active ingredient or ingredients already used.

[0043] The dosage of the drugs used in the present invention is determined by comprehensively considering the pharmaceutical properties of each component to be combined and the properties of the pharmaceutical composition or pharmaceutical formulation. For the application of the pharmaceutical composition or pharmaceutical formulation of the present invention, a therapeutically effective amount of the active ingredient should be provided. For example, if an animal treatment regimen is applied to humans, the effective dose of the active ingredient in humans can be converted from the effective dose of the drug in animals. This is foreseeable and easily achievable by those skilled in the art.

[0044] The "simultaneous administration" of the pharmaceutical composition of the present invention refers to the simultaneous administration of vortioxetine or a pharmaceutically acceptable salt thereof and olanzapine.

[0045] As used herein, when referring to numerical ranges, unless otherwise specified, the numerical ranges are considered continuous and include the minimum and maximum values of the range, as well as every value between such minimum and maximum values. Furthermore, when a range refers to an integer, every integer between the minimum and maximum values of the range is included. Unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges subsumed therein.

[0046] The dosage of experimental animals (C57 mice) involved in this article has been converted from the clinical dose mg / day (based on an adult weight of 60kg) to mg / kg (for the convenience of comparison, the unit dose of the marketed drug Symbyax is benchmarked, with the two active ingredients in contents of 50mg / day and 6mg / day). The corresponding dosage for mice is obtained according to the conversion formula. Because the weight of individual C57 mice varies, listing the oral administration dose for each C57 mouse one by one is not concise. For the convenience of description, this article uses the human clinical dose instead when describing the oral administration dose for mice, and the oral administration in this article is all one-time oral administration. The dosage conversion formula between humans and animals known to those skilled in the art is as follows:

[0047] Animal dose (mg / kg body weight) = human dose (mg / kg body weight) × human Km / animal Km

[0048] The calculation method is based on body surface area per square meter (BSA, m 2 ) weight (kg), that is, the unit of the conversion factor Km is kg / m 2 For conversion, the Km of different animals is different. In this article, the Km of adults is 36.9, and the Km of mice is 3.

[0049] In a biological experiment, the present invention used vortioxetine hemihydrobromide and olanzapine in combination to administer the drug, and verified that the beneficial effect of the combined administration of the two active ingredients was superior to that of individual drugs and the marketed drug Symbyax. Vortioxetine hemihydrobromide and olanzapine were then prepared into a compound tablet. The prepared compound tablet has good dissolution, stability and content uniformity. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 : Characterization of OD (450nm) values of cell damage by D-GAL and recovery of damaged cells after administration of different drugs DETAILED DESCRIPTION

[0051] The above contents of the present invention are further described in detail below through specific embodiments, but this should not be construed as limiting the present invention.

[0052] Sources and specifications of materials and animals used in this invention:

[0053] Experimental reagents: pH = 4.5 acetic acid-sodium acetate buffer, 0.5% CMC-Na (sodium carboxymethyl cellulose)

[0054] Experimental equipment: balance, mouse gavage needle, camera, all purchased from the market

[0055] Experimental animals: C57 mice, 6-8 weeks old, 20-25 g, male, purchased from Zhejiang Weitonglihua Experimental Animal Technology Co., Ltd.

[0056] Test drugs: Olanzapine and pharmaceutically acceptable salts of vortioxetine were provided by Puji Biotechnology (Taizhou) Co., Ltd.; fluoxetine hydrochloride was purchased from McLean

[0057] The "room temperature" herein refers to 25 to 30°C.

[0058] The "each unit preparation" mentioned in the present invention refers to each vial, tablet, capsule or other unit preparation.

[0059] Example 1: Preparation of test drug solution

[0060] 1. Preparation of olanzapine test solution: Weigh an appropriate amount of olanzapine and dissolve it in acetic acid-sodium acetate buffer (pH 4.5) to prepare a clear solution;

[0061] 2. Preparation of vortioxetine hemihydrobromide test solution: Weigh an appropriate amount of vortioxetine hemihydrobromide and dissolve it in acetic acid-sodium acetate buffer (pH 4.5) to prepare a clear solution;

[0062] 3. Preparation of vortioxetine hydrobromide test solution: Weigh an appropriate amount of vortioxetine hydrobromide and dissolve it in acetic acid-sodium acetate buffer (pH 4.5) to prepare a clear solution;

[0063] 4. Preparation of vortioxetine hemihydrobromide + olanzapine compound solution: Weigh appropriate amounts of vortioxetine hemihydrobromide and olanzapine and dissolve them in acetic acid-sodium acetate buffer (pH 4.5) to prepare a clear solution;

[0064] 5. Preparation of vortioxetine hydrochloride test solution: Weigh an appropriate amount of vortioxetine hydrochloride and dissolve it in acetic acid-sodium acetate buffer (pH 4.5) to prepare a clear solution;

[0065] 6. Preparation of the compound solution of fluoxetine hydrochloride and olanzapine: Weigh appropriate amounts of fluoxetine hydrochloride and olanzapine and dissolve them in an ethanol aqueous solution (ethanol content is 1%), then add 2% SDS solution to dilute and obtain a clear solution.

[0066] 7. Preparation of Fluoxetine Hydrochloride Solution: Weigh an appropriate amount of Fluoxetine Hydrochloride and dissolve it in ethanol aqueous solution (ethanol content is 1%).

[0067] Then add 2% SDS solution to dilute and obtain a clear solution.

[0068] The concentration of the above test drug solution is adjusted according to the specific experimental requirements. This example is only used to illustrate the preparation method of the test drug solution and the solvent used.

[0069] Example 2: Comparison of gastrointestinal side effects in mice after administration

[0070] Group (1): Six male C57 mice were fasted for 12 hours before administration and then orally administered with 50 mg / day of fluoxetine hydrochloride and 6 mg / day of olanzapine, the active ingredients of Symbyax, in a compound solution prepared according to the method of Example 1. The mice were videotaped throughout the administration process and monitored at 30 minutes, 2 hours, 4 hours, 24 hours, and 72 hours. The mice showed signs of retching, erect hair, and prone position at 30 minutes.

[0071] (2) Groups: Group A, Group B, and Group C. Six male C57 mice were prepared in each group. They were fasted for 12 hours before administration and then orally administered with 50 mg / day of vortioxetine hemihydrobromide and three different doses of olanzapine, prepared as compound solutions according to the method of Example 1. The mice were videotaped throughout the administration process and monitored at 30 minutes, 2 hours, 4 hours, 24 hours, and 72 hours. Table 1 below shows the responses of the three groups of mice at different time points under different compound doses:

[0072] Table 1. Comparison of gastrointestinal side effects in mice

[0073]

[0074]

[0075] It can be concluded from Table 1 that SF001+Y004 (50 mg / day+6 mg / day) had no obvious side effects throughout the entire treatment period, but fluoxetine hydrochloride+Y004 (50 mg / day+6 mg / day) had obvious side effects, indicating that at the same dose, SF001+Y004 can significantly reduce the side effect of dry retching compared with fluoxetine hydrochloride+Y004.

[0076] Example 3: Determination of neurotransmitters in the hippocampus of mice in the compound drug group and the blank group

[0077] Experimental Methods: Male C57 mice were divided into two groups of 10 each: one group was gavaged with an equal volume of normal saline (blank group); the other group was gavaged with SF001 + Y004 (clinical dose 30 mg / day + 15 mg / day). They were fasted for 12 hours before the experiment and given the drug by gavage for three consecutive days. On the fourth day, the mice were killed by cervical dislocation. The brain tissue was quickly dissected on ice, and the hippocampus was isolated and quickly frozen in liquid nitrogen. Neurotransmitter concentrations were detected using LC-MS (liquid chromatography-mass spectrometry). The LC-MS detection method is as follows:

[0078] a. Weigh 50 mg of hippocampal brain sample from each mouse and place it in a centrifuge tube; b. Add 400 uL of 50% methanol and magnetic beads for disruption; c. Centrifuge at 25,000 rpm for 2 minutes and set aside; d. Prepare the standard curve: Take a mixed solution of 39 neurotransmitter standards and perform a gradient dilution; e. Take 20 uL of sample and standard curve, add 60 uL of pre-cooled 50% methanol, shake for 5 minutes, precipitate at -20°C for 4 hours, centrifuge at 20,000g, 4°C for 15 minutes, and take 20 uL of the supernatant to an EP tube; f. Derivatization: Add 20 uL of 200 mM 3-NPH and 20 uL of 200 mM EDC-6% pyridine mixture to the EP tube containing 20 uL of supernatant, incubate at 25°C with shaking for 30 minutes, centrifuge, take the supernatant, load onto the analyzer, and detect.

[0079] The above-mentioned detection experiment was conducted using a Waters Class I-AB Sciex 6500 LC / MS instrument with a Waters BEH C18 column (model: 1.7 μm x 2.1 μm x 100 mm). Mobile phase A consisted of water + 0.1% formic acid, and mobile phase B consisted of methanol + 0.1% formic acid; the flow rate was 0.35 mL / min; the gradient was set as 2% B (0-2 min), 20-80% B (2.5-15 min).

[0080] Experimental results: The results of the neurotransmitter measurement in the hippocampus are shown in Table 2 below:

[0081] Table 2. Results of neurotransmitter measurements in the mouse hippocampus

[0082]

[0083] Note: t.test_p.value, that is, P value < 0.05, indicates that there is a statistical difference between the groups.

[0084] In the article "Modulation of glutamate receptor functions by glutathione. Neurochemistry International 37, 299-306, 2000," it is pointed out that glutathione also plays a dual role as a neurotransmitter and neuromodulator in the central nervous system. Neurochemical, neuropharmacological and electrophysiological studies have all demonstrated these two functions of glutathione. As an excitatory neurotransmitter, glutathione depolarizes neurons through ion conduction, depolarizing neurons by acting as its own ion receptor (unlike any other excitatory amino acid receptor). As a neuromodulator, it can remove ionotropic glutamate receptor ligands from their binding sites and regulate the influx of calcium ions through ion channels controlled by N-methyl-D-aspartate receptors. In brain slices, glutathione has been shown to regulate the release of other neurotransmitters, for example, glutathione can regulate other neurotransmitters through N-methyl-D-aspartate receptors. During antidepressant treatment, it is found that certain neurotransmitter metabolites increase, such as normetanephrine (NMN), an intermediate metabolite of NE, and HVA, a metabolite of DA. The decrease in norepinephrine (NE) levels is considered to be the main pathogenic factor of depression. As shown in Table 2, after treatment with SF001+Y004, the concentrations of GSH, NE, NEM, and HVA neurotransmitters increased significantly compared with the blank group (P < 0.05), indicating that the SF001+Y004 dosing regimen for treating depression has potential efficacy. Example 4: Determination of neurotransmitters in the hippocampus of mice in the SF001 group and the SF001+Y004 group

[0085] Experimental Methods: Male C57 mice were divided into two groups of 10 each: one group was gavage-administered SF001 (clinical dose 30 mg / day); the other group was gavage-administered SF001 + Y004 (clinical dose 30 mg / day + 15 mg / day). They were fasted for 12 hours before the experiment and given oral administration for three consecutive days. On the fourth day, the mice were killed by cervical dislocation. The brain tissue was quickly dissected on ice, and the hippocampus was isolated and quickly frozen in liquid nitrogen. Neurotransmitter concentrations were detected using LC-MS (liquid chromatography-mass spectrometry). The LC-MS detection method is as follows:

[0086] a. Weigh 50 mg of hippocampal brain sample from each mouse and place it in a centrifuge tube; b. Add 400 uL of 50% methanol and magnetic beads for disruption; c. Centrifuge at 25,000 rpm for 2 minutes and set aside; d. Prepare the standard curve: Take a mixed solution of 39 neurotransmitter standards and perform a gradient dilution; e. Take 20 uL of sample and standard curve, add 60 uL of pre-cooled 50% methanol, shake for 5 minutes, precipitate at -20°C for 4 hours, centrifuge at 20,000g, 4°C for 15 minutes, and take 20 uL of the supernatant to an EP tube; f. Derivatization: Add 20 uL of 200 mM 3-NPH and 20 uL of 200 mM EDC-6% pyridine mixture to the EP tube containing 20 uL of supernatant, incubate at 25°C with shaking for 30 minutes, centrifuge, take the supernatant, load on the analyzer, and detect.

[0087] The above-mentioned detection experiment was conducted using a Waters Class I-AB Sciex 6500 LC / MS instrument with a Waters BEH C18 column (model: 1.7 μm x 2.1 μm x 100 mm). Mobile phase A consisted of water + 0.1% formic acid, and mobile phase B consisted of methanol + 0.1% formic acid; the flow rate was 0.35 mL / min; the gradient was set as 2% B (0-2 min), 20-80% B (2.5-15 min).

[0088] Experimental results: The results of the neurotransmitter measurement in the hippocampus are shown in Table 3 below:

[0089] Table 3. Results of neurotransmitter measurements in the mouse hippocampus

[0090]

[0091] 5-HT is a tryptophan metabolite, and Tryptophol is a metabolite of tryptophan in another pathway. The two are in a competitive relationship. Therefore, when the concentration of 5-HT in the brain increases, the concentration of Tryptophol, another competitive metabolite of tryptophan, decreases. N-Acetylserotonin is a reaction intermediate in the endogenous synthesis of 5-HT to melatonin. DOPAC (dihydroxyphenylacetic acid), HVA and Vanillmandelic_acid are dopamine metabolites. L-Dopa is a dopamine precursor. NE and Normetanephrine are dopamine metabolites.

[0092] From the data in Table 3, it can be concluded that when the combination of SF001 and Y004 was administered to mice, the concentrations of 10 neurotransmitters in the hippocampus were upregulated and the concentration of 1 neurotransmitter was downregulated compared to the administration of SF001 alone. Among them, the concentrations of 5-HT, dopamine (DA) and norepinephrine (NE) were significantly increased, indicating that the combined administration of SF001 and Y004 had a synergistic effect. This enhanced synergistic effect of SF001 and Y004 is also an intuitive evidence that the SF001 + Y004 combination has a faster onset and better efficacy in treating depression, anxiety and improving cognitive impairment.

[0093] Example 5: Determination of neurotransmitters in the hippocampus of mice in the SF001+Y004 group and the fluoxetine+Y004 group

[0094] Experimental methods: Male C57 mice were divided into two groups of 10 each. One group was gavage-administered SF001+Y004 (clinical dose 30 mg / day+15 mg / day), and the other group was gavage-administered fluoxetine+Y004 (clinical dose 30 mg / day+15 mg / day). The mice were fasted but not watered for 12 hours before the experiment. The mice were gavage-administered for 3 consecutive days. On the 4th day, the mice were killed by cervical dislocation, and the brain tissue was quickly peeled off on ice. The hippocampus was isolated and quickly frozen in liquid nitrogen. The neurotransmitter concentrations were detected by LC-MS (liquid chromatography-mass spectrometry).

[0095] Among them, the LC-MS detection method is:

[0096] a. Weigh 50 mg of hippocampal brain sample from each mouse and place it in a centrifuge tube; b. Add 400 uL of 50% methanol and magnetic beads for disruption; c. Centrifuge at 25,000 rpm for 2 minutes and set aside; d. Prepare the standard curve: Take a mixed solution of 39 neurotransmitter standards and perform a gradient dilution; e. Take 20 uL of sample and standard curve, add 60 uL of pre-cooled 50% methanol, shake for 5 minutes, precipitate at -20°C for 4 hours, centrifuge at 20,000g, 4°C for 15 minutes, and take 20 uL of the supernatant to an EP tube; f. Derivatization: Add 20 uL of 200 mM 3-NPH and 20 uL of 200 mM EDC-6% pyridine mixture to the EP tube containing 20 uL of supernatant, incubate at 25°C with shaking for 30 minutes, centrifuge, take the supernatant, load on the analyzer, and detect.

[0097] The above-mentioned detection experiment was conducted using a Waters Class I-AB Sciex 6500 LC / MS instrument with a Waters BEH C18 column (model: 1.7 μm x 2.1 μm x 100 mm). Mobile phase A consisted of water + 0.1% formic acid, and mobile phase B consisted of methanol + 0.1% formic acid; the flow rate was 0.35 mL / min; the gradient was set as 2% B (0-2 min), 20-80% B (2.5-15 min).

[0098] Table 4: Results of neurotransmitter measurements in the hippocampus

[0099]

[0100] Compared with the administration of the fluoxetine + Y004 combination, the melatonin content decreased after the administration of the SF001 + Y004 combination, which can offset the side effect of drowsiness caused by taking Y004 to a certain extent. In the document Identification of a Novel Strain Lactobacillus Reuteri and Anti-Obesity Effect through Metabolite Indole-3-Carboxaldehyde in Diet-Induced Obese Mice., Journal of Agricultural and Food Chemistry., 2023, 71(7), 3239-3249, it is pointed out that indole and its derivatives, as the main metabolites of tryptophan, play an important role in reducing inflammation, regulating immunity, and inhibiting obesity. The abundance of indole-3-carbaldehyde is negatively correlated with obesity-related indicators. It mainly participates in the anti-obesity mechanism by regulating the cGMP / cAMP signaling pathway and inhibiting lipid deposition. Compared with the administration of the fluoxetine + Y004 combination, the indole-3-carboxaldehyde content was reduced after the administration of the SF001 + Y004 combination, which alleviated the obesity-related side effect of taking Y004 to a certain extent.

[0101] Example 6:

[0102] The experimental materials, experimental cells, and experimental equipment are shown in Table 5 below:

[0103] Table 5. Experimental materials, experimental cells, and experimental equipment

[0104]

[0105]

[0106] Experimental steps:

[0107] (1) Take out the 15% FBS DMEM high glucose medium from the refrigerator in advance and return it to room temperature. Irradiate with UV light in the biosafety cabinet for 30 minutes. (2) Inoculate 96-well plates: Digested SH-SY5Y cells were plated at a rate of 2.5×10 5 The cells were seeded into 96-well plates at a density of 100 μg / mL, with 3 blank wells on each plate. 200 μL of PBS buffer was added to each well around the plate. The cells were gently shaken to mix and then placed in a CO2 incubator for culture.

[0108] (3) Preparation of D-galactose (D-GAL) solution: 0.8 g D-galactose was added to 800 μL H2O, shaken and mixed, and then heated in a 60°C water bath to dissolve to a concentration of 1000 g / L.

[0109] (4) Drug administration: 12 hours after SH-SY5Y cells (human neuroblastoma cells) were plated, the culture medium in the 96-well plate was removed, and the pre-prepared D-GAL was diluted to 60 g / L and added to the 96-well plate (no D-GAL was added to the normal control group); the drug SF001 (1uM, 100nM, 10nM, 1nM) at the corresponding concentration was added to the drug group 1, and the drug Y004 (120nM, 12nM, 1.2nM, 0.12nM) at the corresponding concentration was added to the drug group 2 and incubated for 24 hours; the drug group 3 was added with the corresponding concentrations of SF001+Y004 (1uM+120nM; 100nM+12nM; 10nM+1.2nM; 1nM+0.12nM) and incubated for 24 hours;

[0110] (5) CCK-8 assay for cell viability: Prepare a CCK-8 mixed solution (i.e., 5 mL of 15% FBS-containing DMEM + 0.5 mL of CCK-8 solution) in advance; remove the culture medium from the wells and wash once with 100 μL of PBS buffer. Add 110 μL of the CCK-8 mixed solution to each well. Incubate in a CO2 incubator for 2.5 h; measure the absorbance (OD value) of the corresponding wells using a microplate reader at a wavelength of 450 nm.

[0111] Experimental results:

[0112] Compared with the normal control group, the cell activity in the D-GAL group was significantly decreased, indicating that the cell aging damage model was successfully established. Compared with the D-GAL group, the cell activity increased to varying degrees after the addition of SF001 (1μM, 100nM) to group 1, Y004 (120nM, 12nM) to group 2, and SF001+Y004 (1μM+120nM, 100nM+12nM) to group 3. The activity of group 3 was significantly higher than that of groups 1 and 2, with statistical differences (P<0.05). Figure 1 In the normal control group, cells grew well and had normal morphology. However, 24 hours after addition of D-GAL (60g / L), significant cell death and abnormal morphology (rounding, detachment, and lack of distinct dendrites and axons) were observed. Addition of SF001 (1μM, 100nM) and Y004 (120nM, 12nM) increased the number of cells attached to the wall, and their morphology was partially restored. Addition of SF001+Y004 significantly increased the number of cells attached to the wall, and their morphology was partially restored. This suggests that SF001+Y004 can help enhance neuronal activity and improve synaptic morphology, potentially improving cognitive impairment.

[0113] Example 7:

[0114] Because Parkinson's disease is caused by a decrease in dopaminergic neurons, SH-SY5Y dopaminergic cells are a typical cell model for studying Parkinson's disease. Previous studies have shown that treatment with MPP+ (1-methyl-4-phenylpyridinium ion) reduces cell survival as the concentration of MPP+ increases. In this study, the SH-SY5Y dopaminergic cell line, which undergoes MPP+-mediated damage, was used as a cell model for studying Parkinson's disease (PD).

[0115] Experimental steps:

[0116] (1) Take out the 15% FBS DMEM high glucose medium from the refrigerator in advance and return it to room temperature. Irradiate with UV light in the biosafety cabinet for 30 minutes. (2) Inoculate 96-well plates: Digested SH-SY5Y cells were plated at a rate of 2.5×10 5 The cells were seeded into 96-well plates at a density of 100 μg / mL, with 3 blank wells on each plate. 200 μL of PBS buffer was added to each well around the plate. The cells were gently shaken to mix and then placed in a CO2 incubator for culture.

[0117] (3)MPP + Dissolve in DMSO (dimethyl sulfoxide) to a final working concentration of 1.5 mM, and store in aliquots at -20°C.

[0118] (4) Administration: 12 hours after SH-SY5Y cells were plated, the culture medium in the first well of the administration group was removed and SF001 (100nM, 10nM, 1nM) was added for pretreatment for 24h, and then the pre-prepared MPP was added. + (No treatment was added in the normal control group) for 24 hours; in the treatment group 2, corresponding concentrations of SF001+Y004 (100nM+12nM; 10nM+1.2nM; 1nM+0.12nM) were added.

[0119] (5) CCK-8 assay for cell viability: Prepare a CCK-8 mixed solution (i.e., 5 mL of 15% FBS-containing DMEM + 0.5 mL of CCK-8 solution) in advance; remove the culture medium from the wells and wash once with 100 μL of PBS buffer. Add 110 μL of the CCK-8 mixed solution to each well. Incubate in a CO2 incubator for 2.5 h; measure the absorbance (OD value) of the corresponding wells using a microplate reader at a wavelength of 450 nm.

[0120] Experimental results:

[0121] MPP +Compared with the normal control group, the cell activity of the group was significantly decreased, indicating that the cell PD injury model was successfully established; and MPP + Compared with the control group, the cell activity of group 1 after adding SF001 (10nM) and group 2 after adding SF001+Y004 (10nM+1.2nM) increased significantly, but the cell activity of group 2 was significantly higher than that of group 1, with a statistically significant difference (P<0.05). This shows that SF001+Y004 can help improve the activity of nerve cells and has the potential to improve cognitive impairment.

[0122] Example 8

[0123]

[0124] Tablet preparation: vortioxetine hemihydrobromide, mannitol, hydroxypropyl cellulose, and magnesium stearate are passed through a 40-mesh sieve; wet granulation is performed: 0.08 g of hydroxypropyl cellulose is added to 1.75 g of purified water and stirred to dissolve; 1 / 3 by weight of mannitol, vortioxetine hemihydrobromide, olanzapine, the remaining mannitol, and the remaining hydroxypropyl cellulose are sequentially mixed and stirred; after stirring, an aqueous solution of hydroxypropyl methylcellulose is added to the mixture, and the mixture is stirred again, granulated, dried, and sized; cross-linked polyvinylpyrrolidone and magnesium stearate are added, mixed, and tableted to obtain vortioxetine hemihydrobromide and olanzapine combination tablets weighing 120 mg.

[0125] Example 9

[0126]

[0127] The above tablet preparation method is referred to the tablet preparation method of Example 8 to obtain a compound tablet with a tablet weight of 120 mg.

[0128] Example 10

[0129]

[0130]

[0131] The above tablet preparation method is referred to the tablet preparation method of Example 8 to obtain a compound tablet with a tablet weight of 120 mg.

[0132] Example 11

[0133]

[0134] The above tablet preparation method is referred to the tablet preparation method of Example 8 to obtain a compound tablet with a tablet weight of 120 mg.

[0135] Example 12

[0136]

[0137] The above tablet preparation method is referred to the tablet preparation method of Example 8 to obtain a compound tablet with a tablet weight of 60 mg.

[0138] Test Example 1 Determination of tablet dissolution curve

[0139] The dissolution and release rate determination method 0931 of the 2020 edition of the Chinese Pharmacopoeia was used, and the second method (paddle method) was used for detection. The dissolution medium was 0.1N HCl solution, the volume was 900 mL, the rotation speed was 50 r / min, and the sampling time was 5, 10, 15, 20, and 30 min. The drug content was determined by ultraviolet detection, and the dissolution rate was calculated. The results are shown in the table below:

[0140] Table 6 Dissolution curve measurement results of vortioxetine hemihydrobromide and olanzapine compound tablets

[0141]

[0142] As can be seen from the above table, the dissolution amount of the compound tablets prepared in the present application in 0.1N HCl solution within 15 minutes is greater than 85%, which proves that the tablets of the present application can achieve rapid dissolution after disintegration and dispersion.

[0143] Test Example 2 Stability Investigation

[0144] The compound tablets prepared in Examples 8 to 12 of the present application were stored sealed for 6 months and unsealed for 6 months at 40°C ± 2°C / 75% RH ± 5% RH, respectively. Test results showed that the maximum single impurity content ranged from 0.023% to 0.056%, the total impurity content of SF001 ranged from 0.016% to 0.084%, and the total impurity content of Y004 ranged from 0.091% to 0.447%.

[0145] Test Example 3 Gastrointestinal side effects in mice after administration

[0146] Referring to the experimental method of Example 2: Take one tablet from each of Examples 8-12, crush it, and add it to water to obtain a solution. Prepare five groups of male C57 mice, each with six mice. Fast for 12 hours before administration, but not water. The C57 mice were gavage-administered with solutions of the tablets from Examples 8-12. The entire process of administration was videotaped, and the mice were monitored at 30 minutes, 2 hours, 4 hours, 24 hours, and 72 hours. The mice showed no obvious side effects throughout the entire process and behaved normally. Based on the experimental results, the vortioxetine hemihydrobromide and olanzapine compound tablets prepared in this application have essentially no gastrointestinal side effects.

[0147] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not mean that the essence of the corresponding technical solutions deviates from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A pharmaceutical composition, characterized in that Contains two active ingredients: (1) vortioxetine or a pharmaceutically acceptable salt thereof; and (2) Olanzapine; The weight ratio of vortioxetine or a pharmaceutically acceptable salt thereof to olanzapine is 6-50:3-6, preferably 9-50:6, and the weights of the two active ingredients are calculated in free form.

2. The pharmaceutical composition according to claim 1, characterized in that The weight ratio of vortioxetine or a pharmaceutically acceptable salt thereof to olanzapine is 9:6, 10:6, 12:6, 15:6, 18:6, 20:6, 22:6, 25:6, 27:6, 30:6, 35:6, 40:6, 45:6 or 50:

6.

3. The pharmaceutical composition according to any one of claims 1 to 2, characterized in that The pharmaceutically acceptable salt is a salt formed by vortioxetine and hydrobromic acid, hydrochloric acid, methanesulfonic acid, fumaric acid, maleic acid, tartaric acid, sulfuric acid, phosphoric acid or nitric acid.

4. The pharmaceutical composition according to claims 1 to 3, characterized in that The pharmaceutically acceptable salt of vortioxetine is vortioxetine hydrobromide or vortioxetine hemihydrobromide, preferably vortioxetine hemihydrobromide.

5. The pharmaceutical composition according to any one of claims 1 to 4, characterized in that The combination of the two active ingredients is vortioxetine hemihydrobromide and olanzapine, or vortioxetine hydrobromide and olanzapine, preferably a combination of vortioxetine hemihydrobromide and olanzapine.

6. The pharmaceutical composition according to any one of claims 1 to 5, characterized in that The two active ingredients are administered by combined administration, which is simultaneous administration or sequential administration, preferably simultaneous administration.

7. A pharmaceutical preparation, characterized in that The pharmaceutical preparation is composed of a therapeutically effective amount of the pharmaceutical composition according to any one of claims 1 to 6 and a pharmaceutically acceptable carrier.

8. The pharmaceutical preparation according to claim 7, characterized in that The pharmaceutical preparation is a tablet, capsule, powder, granule, emulsion, suspension or injection, preferably a tablet.

9. The pharmaceutical preparation according to claim 7 or 8, characterized in that The weight ratio of vortioxetine or a pharmaceutically acceptable salt thereof to olanzapine in each unit preparation of the pharmaceutical preparation is 5mg:1.5mg, 9mg:6mg, 9mg:3mg, 10mg:3mg, 12mg:6mg, 15mg:6mg, 18mg:6mg, 20mg:6mg, 22mg:6mg, 25mg:6mg, 27mg:6mg, 30mg:6mg, 35mg:6mg, 40mg:6mg, 45mg:6mg or 50mg:6mg.

10. The pharmaceutical preparation according to claim 8 or 9, characterized in that The pharmaceutical preparation contains one or more fillers, one or more binders, one or more disintegrants, and one or more lubricants, wherein the filler is selected from at least one of mannitol, microcrystalline cellulose, lactose, starch, and sorbitol; the binder is selected from at least one of hydroxypropyl cellulose, ethyl cellulose, and copovidone; the disintegrant is selected from at least one of cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium starch glycolate, and low-substituted hydroxypropyl cellulose; and the lubricant is selected from at least one of talc, magnesium stearate, and sodium stearyl fumarate.

11. The pharmaceutical preparation according to claim 10, characterized in that The pharmaceutical preparation contains: 。 12. The pharmaceutical preparation according to claim 11, characterized in that The pharmaceutical preparation contains: 。 13. Use of the pharmaceutical composition of any one of claims 1 to 7 or the pharmaceutical formulation of any one of claims 8 to 12 in the preparation of a medicament for treating a disease, wherein the disease is a mood disorder, depression, anxiety, stress urinary incontinence, vomiting, IBS (irritable bowel syndrome), eating disorders, chronic pain, Alzheimer's disease, ADHD (attention deficit hyperactivity disorder), PTSD (post-traumatic stress disorder), hot flashes, sleep apnea, alcoholism, nicotine or carbohydrate addiction, drug abuse, alcohol or drug abuse, major depressive disorder, postpartum depression, depression associated with bipolar disorder, psychosis, cancer, aging or Parkinson's disease, generalized anxiety disorder, social anxiety disorder, obsessive-compulsive disorder, panic disorder, panic attacks, phobia, social phobia, agoraphobia, partial response, resistant depression, cognitive impairment or dysthymia, preferably a mood disorder, depression, anxiety, resistant depression or cognitive impairment.

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