WO2023237124A1 - 一种藏红花素混悬剂及其在制备快速抗抑郁药物中的应用 - Google Patents

一种藏红花素混悬剂及其在制备快速抗抑郁药物中的应用 Download PDF

Info

Publication number
WO2023237124A1
WO2023237124A1 PCT/CN2023/102757 CN2023102757W WO2023237124A1 WO 2023237124 A1 WO2023237124 A1 WO 2023237124A1 CN 2023102757 W CN2023102757 W CN 2023102757W WO 2023237124 A1 WO2023237124 A1 WO 2023237124A1
Authority
WO
WIPO (PCT)
Prior art keywords
crocin
suspension
mice
suspending agent
rapid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2023/102757
Other languages
English (en)
French (fr)
Inventor
王广基
阿基业
张悦
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
China Pharmaceutical University
Original Assignee
China Pharmaceutical University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by China Pharmaceutical University filed Critical China Pharmaceutical University
Publication of WO2023237124A1 publication Critical patent/WO2023237124A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7024—Esters of saccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A61K47/38—Cellulose; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/24—Antidepressants

Definitions

  • the invention belongs to the field of medicine, and relates to crocin suspension and its application in preparing rapid antidepressant drugs.
  • it relates to crocin suspension regulating the level of cyclic adenosine monophosphate (cAMP) in the nucleus accumbens brain area through the vagus nerve, exerting rapid , safe, efficient antidepressant effect and its application.
  • cAMP cyclic adenosine monophosphate
  • depression has become the second largest killer of humans after cancer.
  • the global average incidence of depression is about 4.4%, and in the past 10 years, the number of patients has increased by more than 18%.
  • the lifetime prevalence rate of depression in my country is 6.9%, and the 12-month prevalence rate is 3.6%. Based on this, it is estimated that the There are nearly 100 million people suffering from depression.
  • the consultation rate of patients with depression in my country is only 9.5%, and among the patients who seek treatment, only 0.5% receive adequate treatment.
  • ketamine a drug
  • ketamine can relieve depressive symptoms in 3-4 hours after a single injection.
  • antidepressant mechanism of ketamine is still unclear and it has strong addictive properties. It cannot be used on a large scale, but due to its precise antidepressant effect, the United States and the European Union have successively approved the marketing of esketamine for the treatment of "treatment-resistant depression.”
  • psilocybin another rapid antidepressant, psilocybin (from psychedelic mushrooms, a hallucinogen, which belongs to the category of drugs).
  • crocin can regulate cAMP levels in the nucleus accumbens and significantly improve depression-like behavior in experimental animals.
  • Intravenous injection of crocin showed no obvious effect, and the effect disappeared after cutting off the vagus nerve connecting the intestines and the central nervous system. It was demonstrated that the efficacy of oral crocin must depend on its entry into the intestinal tract. Further research also found that higher doses of crocin can accumulate in the cecum and colon through oral administration. In the intestine, it can activate the vagus nerve and increase cAMP levels in the nucleus accumbens brain area through neural circuit regulation, exerting a rapid antidepressant effect.
  • crocin has a mechanism for exerting rapid antidepressant effects through the intestines. It not only avoids the problem that drugs need to enter the brain through the blood-brain barrier, but also uses the body's own neural circuit regulation to specifically act on the nucleus accumbens brain area. It also has It has the characteristics of rapid onset of action and good safety, and the oral administration has good patient compliance. Preparing crocin into a preparation that is conducive to its rapid antidepressant effect in the intestines. After oral administration, it can quickly relieve patients' depressive symptoms within 1-2 days, which has extremely important clinical significance and broad market prospects.
  • crocin has low solubility under ordinary physiological environments, weak permeability of biofilms, instability in acidic and alkaline environments, and is easily degraded into other substances and reduce the efficacy of the medicine. Since ordinary tablets and capsules enter the stomach after oral administration and gradually disintegrate in the stomach, crocin and excipients will remain in the folds of the stomach for a long time and adhere to the gastric wall, causing damage in the acidic environment of the stomach and directly Reducing the amount of drug that enters the intestine to exert its effect is not conducive to the effective use of the drug.
  • the purpose of the present invention is to provide crocin in the preparation of suspensions and its application in rapid antidepressant drugs in view of the lack of existing depression treatment drugs and the above-mentioned deficiencies in technology.
  • a crocin suspension characterized by containing the following mass ratio of active ingredients and excipients: crocin: suspending agent ratio is 0.2-20:0.4-2, and the balance is water, in which the suspending agent and the suspending agent The mass percentage of the agent is 0.2-0.5%.
  • crocin exerts rapid antidepressant effects by activating the vagus nerve in the intestine and regulating cAMP levels in the nucleus accumbens through the gut-brain circuit.
  • crocin needs to be activated in the cecum to exert its effects through this mechanism.
  • CMC-Na sodium carboxymethylcellulose
  • HPMC hydroxypropyl methylcellulose
  • crocin of the present invention is as follows:
  • the suspending agent of the present invention is selected from the group consisting of Avicel CL611, Avicel CL591, hydroxypropyl cellulose, hydroxyethyl cellulose, sodium carboxymethyl cellulose, ethyl cellulose, methyl cellulose, xanthan Glue, sodium alginate, polyvinyl alcohol, povidone k-30, povidone k-90, polyvinylpyrrolidone.
  • the suspending agent is sodium carboxymethyl cellulose.
  • the mass ratio of crocin:sodium carboxymethylcellulose per unit suspension is 3-20:0.4-1.
  • the mass ratio of crocin:carboxymethylcellulose sodium per unit suspension is 4-8:1.
  • the mass ratio of crocin:carboxymethylcellulose sodium per unit suspension is 6:1.
  • the crocin suspension is simply physically mixed with the active ingredient crocin and the suspending agent, and then dissolved in water to form a suspension; or the active ingredient crocin is dispersed In the pre-prepared aqueous solution of suspending agent, it becomes a suspension.
  • each preparation contains crocin at a concentration of 1-100 mg/mL, preferably 15-30 mg/mL.
  • the dosage of crocin suspension for human administration is 0.5-4g/60kg based on crocin, preferably 1-2g/60kg.
  • the unit dose of crocin is 50-400 mg based on crocin.
  • the unit dose of crocin in the suspension is 100-200 mg calculated as crocin; the aforementioned high-dose crocin can be displayed within 24 hours-48 hours after a single administration Obvious rapid antidepressant effect.
  • the dosage form of the pharmaceutical preparation includes a dosage form for oral administration.
  • the unit dose of crocin is 50-400 mg based on crocin.
  • the unit dose of crocin in the crocin oral suspension is 100-200 mg calculated as crocin; the aforementioned crocin oral suspension is administered in a single oral administration within 24 hours - It can show obvious rapid antidepressant effects within 48 hours.
  • the volume of crocin suspension for human administration is 5-50 mL, preferably 10-20 mL, based on the suspension.
  • crocin suspension when the active ingredient crocin is significantly accumulated in the cecum and colon, it exerts a rapid antidepressant effect in the intestine by increasing the cAMP level in the nucleus accumbens through vagus nerve regulation.
  • cyclic adenosine monophosphate cAMP
  • cAMP cyclic adenosine monophosphate
  • crocin suspension accumulates in the intestine after oral administration, and exerts a rapid antidepressant effect by increasing cAMP levels in the nucleus accumbens through vagus nerve regulation. Cutting off the vagus nerve blocks crocin from increasing the level of cAMP in the nucleus accumbens.
  • the sources of crocin in the present invention include, but are not limited to, the Rubiaceae plant Gardenia grandiflora, the fruits of Gardenia, the Iris plant saffron and the stigmas of other plants of the same genus, the Strychnaceae plant Buddleia japonica, and the Oleaceae plant night flowers.
  • burdock, asteraceae plant, mimosa plant, leguminous plant, mimosa, etc. are used as raw materials, and are extracted using modern biotechnology, as well as other artificially synthesized crocin.
  • Saffron is a traditional and valuable traditional Chinese medicine. It is non-toxic. It is used for depression, persistent shortness of breath, and activating blood circulation. Eating it for a long time makes people feel happy.
  • crocin in the traditional Chinese medicine saffron can increase cAMP levels in the nucleus accumbens brain area by activating the vagus nerve, and quickly improves depressive symptoms 24-48 hours after administration. Its oral efficacy is comparable to intraperitoneal injection of ketamine.
  • the rapid antidepressant effects of crocin were demonstrated in three recognized depression models, in three independent laboratories, under double-blind conditions. In addition, studies have found that intravenous injection of a considerable amount of crocin is ineffective.
  • crocin suspension When crocin suspension accumulates in the intestine after oral administration, it exerts a rapid antidepressant effect by increasing cAMP levels in the nucleus accumbens through vagus nerve regulation. Cutting off the vagus nerve blocks the rise of crocin. High accumbens cAMP and counteracts its antidepressant effects. This shows that crocin is a new, rapid antidepressant drug that has a unique ability to exert a rapid antidepressant effect by regulating the cAMP in the nucleus accumbens through the intestinal vagus nerve and neural circuits. It has the characteristics of fast onset and good safety. Moreover, oral administration has better patient compliance. Crocin suspension can effectively accumulate a sufficient amount of crocin in the intestine, thereby exerting antidepressant effects through a new mechanism.
  • the crocin suspension prepared by the process of the present invention is beneficial to reducing degradation in gastric acid environment.
  • the crocin suspension is in a suspension state with a certain viscosity. After oral administration, the liquid will not only be discharged from the stomach faster than the solid particles Empty into the intestinal tract, and most of the particles in the suspension are dispersed in a solution with a certain viscosity, reducing contact with acidic gastric juice and direct exposure in the stomach.
  • the above-mentioned dual factors also cause the crocin in the suspension to be less affected and destroyed by gastric juice, which increases the amount of drug entering the intestine and is more conducive to exerting its medicinal effect in the intestine.
  • the present invention clarifies that crocin exerts antidepressant effects through a new mechanism, fills the gap in international fast, safe, and efficient antidepressant treatment drugs, will effectively support its research and development as an antidepressant drug, and benefit hundreds of millions of people suffering from depression around the world. patients, producing huge economic benefits and far-reaching social benefits.
  • Figure 1 Statistical diagram of crocin concentration in the intestinal contents of each intestinal segment 2h, 6h and 12h after administration of three crocin preparations.
  • FIG. 2 Statistical graph of crocin concentration in the intestinal contents of each intestinal segment 2h, 6h and 12h after intragastric administration of crocin suspension prepared with different suspending agents.
  • A crocin suspension prepared by using sodium carboxymethyl cellulose (CMC-Na) as the suspending agent
  • B saffron prepared by using hydroxypropyl methylcellulose (HPMC) as the suspending agent.
  • crocin suspension; 1, 2, and 3 are respectively 7.5, 15, and 30 mg/mL crocin suspensions prepared with CMC-Na as the suspending agent; 4, 5, and 6 are respectively the crocin suspensions with HPMC as the suspending agent.
  • Crocin suspensions with specifications of 7.5, 15, and 30 mg/mL were prepared.
  • FIG. 3 Statistical graph of crocin concentration in the intestinal contents of each intestinal segment 2h, 6h and 12h after administration of crocin suspensions prepared with different CMC-Na concentrations.
  • A Statistical chart of social coefficient of mice in each group
  • B Statistical chart of forced swimming immobility time of mice in each group
  • 1, 2, and 3 are specifications 7.5, 15, and 15 respectively made with CMC-Na as the suspending agent.
  • A Statistical chart of tail suspension immobility time of mice in each group
  • B Statistical chart of forced swimming immobility time of mice in each group
  • 1, 2, and 3 are specifications 7.5 prepared using CMC-Na as the suspending agent respectively. , 15, 30 mg/mL crocin suspension (0.2 mL/g body weight), ketamine dosage is 10 mg/kg (intraperitoneal injection), analyzed by one-way ANOVA, *p ⁇ 0.05, **p ⁇ 0.01 .
  • A Statistical chart of sugar water preference coefficient of mice in each group
  • B Statistical chart of forced swimming immobility time of mice in each group
  • 3 is a 30 mg/mL crocin suspension prepared with CMC-Na as the suspending agent.
  • dose 0.2mL/g body weight
  • the dose of ketamine was 10mg/kg (intraperitoneal injection)
  • one-way ANOVA was used to analyze, ***p ⁇ 0.001, ****p ⁇ 0.0001.
  • A Immunofluorescence staining diagram of c-fos expression in the nucleus tractus solitarius (NTS) of mice in each group; B: Statistical diagram of cAMP levels in the nucleus accumbens of mice in each group; 1, 2, and 3 are respectively based on CMC-Na.
  • A Statistical graph of social coefficient of mice in each group
  • B Statistical graph of tail-suspended immobility time of mice in each group
  • C Statistical graph of sugar water preference coefficient of mice in each group
  • D Nucleus accumbens brain area of mice in each group cAMP concentration statistical chart
  • blank solvent represents intragastric administration of an equal volume of 0.5% CMC-Na, analyzed by two-way ANOVA, *p ⁇ 0.05, **p ⁇ 0.01, ***p ⁇ 0.001, ****p ⁇ 0.0001, n.s. indicates no statistical difference.
  • Preparation process Take 5.0g of crocin, which (extracted from saffron or gardenia, purchased from Mansite Biotechnology Company, purity 99.07%, batch number MUST-20070213) is passed through a 60-mesh sieve; lactose ( Lactose, MEGGLE, Germany, batch number 101504319) 2.0g passed through an 80 mesh sieve; microcrystalline cellulose (JRS, Germany, batch number 5610264626) 3.0g, passed through an 80 mesh sieve.
  • CMC-Na carboxymethyl cellulose sodium
  • Preparation process Take 0.75g/1.5g/3.0g of crocin and pass through a 60-mesh sieve; 0.5g of sodium carboxymethylcellulose (Sinopharm Group, batch number 20131022) is passed through an 80-mesh sieve. Disperse and dissolve sodium carboxymethylcellulose in 100 mL of distilled water. Disperse and suspend crocin in 0.5% sodium carboxymethyl cellulose aqueous solution before use. Shake and mix evenly. 10 mL of each preparation, new Administer after fresh preparation.
  • HPMC hydroxypropyl methylcellulose
  • Preparation process Take 0.75g/1.5g/3.0g of crocin and pass through a 60-mesh sieve; 0.5g of hydroxypropyl methylcellulose (Anhui Shanhe Pharmaceutical Excipients Co., Ltd., batch number 181231) pass through an 80-mesh sieve. Disperse and dissolve hydroxypropyl methylcellulose in 100 mL of distilled water. Disperse and suspend crocin in 0.5% hydroxypropyl methylcellulose aqueous solution before use. Shake and mix evenly. Each preparation is 10 mL. Prepare freshly. After administration.
  • CMC-Na sodium carboxymethyl cellulose
  • Preparation process Take 3.0g of crocin and pass through a 60-mesh sieve; take 0.2g/1.0g of sodium carboxymethylcellulose and pass through an 80-mesh sieve. Disperse and dissolve sodium carboxymethyl cellulose in 100 mL of distilled water. Disperse and suspend crocin in 0.2%/1.0% sodium carboxymethyl cellulose aqueous solution before use. Shake and suspend evenly. Each preparation is 10 mL. Administer freshly prepared.
  • Suspensions were prepared with different amounts of crocin, using sodium carboxymethylcellulose (CMC-Na) as the suspending agent: (Specifications 9 and 10 are 1.0 and 100mg/mL respectively)
  • Preparation process Take 0.1g/10g of crocin and pass through a 60-mesh sieve; 0.5g of sodium carboxymethylcellulose and pass through an 80-mesh sieve. Disperse and dissolve sodium carboxymethyl cellulose in 100 mL of distilled water. Disperse and suspend crocin in 0.5% sodium carboxymethyl cellulose aqueous solution before use. Shake and suspend evenly. Each preparation is 10 mL. Freshly prepared and given. medicine.
  • Example 2 Intestinal accumulation of crocin in three dosage forms after administration
  • C57BL/6J mice (7-8 weeks) were selected, with 6 mice in each group, and the crocin granules, crocin solution and crocin suspension (specification 3) prepared in Example 1 were orally administered (with crocin (calculated, the dosage is 300 mg/kg), and the intestinal accumulation of different crocin preparations after intragastric administration was investigated.
  • crocin calculated, the dosage is 300 mg/kg
  • the intestinal accumulation of different crocin preparations after intragastric administration was investigated.
  • Take the contents of the jejunum, ileum, cecum and colon at 2h, 6h and 12h after administration add ultrapure water at a ratio of 100mg intestinal contents/1mL water and mix thoroughly, then centrifuge and take the supernatant to measure the concentration of crocin. .
  • the conventional pharmacological dosage of crocin in mice is in the range of 50-300 mg/kg.
  • the use of crocin suspension at higher doses is more conducive to the long-term accumulation of crocin in the cecum and colon contents.
  • the crocin suspension dosage form is more conducive to the long-term accumulation of crocin in the cecum and colon.
  • Example 1 of the present invention Take three crocin preparations prepared in Example 1 of the present invention and observe the changes in appearance properties before and after storage at room temperature. The results showed that the crocin suspension and granules were relatively stable, with no obvious change in properties after one week of storage, while the solution showed obvious stratification after one week of storage, see Table 1.
  • Table 1 Changes of three dosage forms before and after being placed at room temperature for 1 week
  • C57BL/6J mice (7 to 8 weeks old) were selected, with 5 mice in each group, and the crocin suspension of the specifications 123456 prepared in Example 1 (0.2 mL/20g) was administered by gavage. body weight) to examine the intestinal accumulation of different crocin suspensions after intragastric administration.
  • crocin suspensions After intragastric administration, Take the contents of the jejunum, ileum, cecum and colon at 2h, 6h and 12h after administration, add ultrapure water at a ratio of 100mg/1mL and mix thoroughly, then centrifuge the supernatant to measure the crocin concentration.
  • C57BL/6J mice (7-8 weeks) were selected, 5 in each group, and were administered intragastrically.
  • the crocin suspension (0.2 mL/20g body weight) of specifications 378 prepared in Example 1 was used to examine the intestinal accumulation of different crocin preparations after intragastric administration. Take the contents of the jejunum, ileum, cecum and colon at 2h, 6h and 12h after administration, add ultrapure water at a ratio of 100mg/1mL and mix thoroughly, then centrifuge the supernatant to measure the crocin concentration.
  • the present invention also selected other suspending agents for testing.
  • the suspending agents include but are not limited to (1) low molecular suspending agents, such as glycerin, syrup, etc.; (2) polymer suspending agents 1 natural: gum arabic, Tragacanth gum, agar, sodium alginate, white gum, pectin, etc.; 2Synthetic products such as Avicel CL611, Avicel CL591, hydroxypropyl cellulose, hydroxyethyl cellulose, ethyl cellulose, Methylcellulose, xanthan gum, sodium alginate, polyvinyl alcohol, povidone k-30, povidone k-90, polyvinylpyrrolidone, etc.; 3 Silicic acids such as colloidal silica, aluminum silicate, Silica bentonite. The test results found that sodium carboxymethyl cellulose is the most preferred suspending agent.
  • Example 5 Acute toxicity test of crocin suspension
  • Test article Take the crocin suspension of the above specification 3, containing 30mg/mL crocin.
  • mice Take 20 mice weighing 18 to 22g, 10 in each group, and administer 0.2 mL of suspension by gavage, which is equivalent to a dosage of 300 mg/kg in terms of crocin. The mice are continuously administered for 7 days. No deaths or abnormalities were seen.
  • the chronic social defeat stress model is the most widely used mouse model of depression in recent years. This model has been used in the professional field because of its obvious inducement (social defeat stress), high modeling success rate, and obvious phenotypic presentation. widely accepted. Since most depression-related stress has social characteristics, including environmental stress, stressful life events, and interpersonal frustrations, it is social in nature. The chronic social frustration stress model can better simulate the social factors that lead to depression. The real process is highly similar to the characteristics of human depression. In addition, it also has lasting behavioral and biological changes. It is one of the best animal models for studying depression.
  • the chronic social defeat model used CD-1 mice (retired mice, male, 4-6 months old, purchased from Beijing Vitong Lihua Experimental Animal Technology Co., Ltd., weight 48 ⁇ 5g) and C57BL/6J mice (male, 7 ⁇ 8 weeks old, purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd., body weight 20 ⁇ 2g), and were adaptively raised for one week under standard feeding environment (free food and water, alternating day and night, 12 hours each).
  • CD-1 mice were screened continuously for 3 days, and aggressive CD-1 mice were selected for modeling.
  • CD-1 mice and C57 mice were separated by transparent plastic partitions with holes, and C57 mice were placed on the same side of the CD-1 mice every day to accept the challenge.
  • a new CD-1 mouse was challenged every day for 10 minutes each time. After the end of the day, the C57 mouse was placed on the opposite side and continued to receive visual and gustatory stress from the CD-1 mouse for the remaining 24 hours. .
  • Modeling continued for 10 days. On the 11th day, mice with a social coefficient less than 1 (i.e., depression-sensitive mice) were screened based on social avoidance behavior for the test.
  • Dosing method The crocein suspension preparation of specifications 123 prepared in Example 1 (0.2 mL/20 g body weight) was orally administered to depression-sensitive mice, with ketamine as the positive drug (intraperitoneal injection, 10 mg/kg), control group and model group was given an equal volume of 0.5% CMC-Na.
  • mice Social behavior was evaluated 24 hours after administration. Before the experiment, the mice were placed in the behavioral testing room to adapt to the light and temperature for more than 30 minutes. Place the C57 mice in an open field of 40 ⁇ 40 ⁇ 40 cm, and place an empty mesh cage in the social area. Record the activities of the C57 mice within 2.5 minutes. After the end, remove the C57 mice and Clear the open area. During the second period, a new CD-1 mouse was placed in the mesh cage and recorded for the same 2.5 minutes. The social coefficient (SI) was calculated as the ratio of the time that C57 mice entered the social area in the second period to the time that they entered the social area in the first period.
  • SI social coefficient
  • mice in the model group decreased significantly compared with the control group.
  • Crocin of specification 3 and the positive drug ketamine could significantly improve the social coefficient.
  • Crocin in other groups had no significant improvement in social avoidance behavior (see Figure 4A) .
  • ketamine is currently the only highly effective and rapid antidepressant drug on the market (it takes effect within a few hours)
  • the crocin suspension prepared by this patented process has an antidepressant effect close to that of ketamine after oral administration, and has no obvious toxic and side effects. It is expected to become a new generation of highly effective, rapid and low-toxic antidepressants.
  • mice Use the forced swimming test (FST) to evaluate the desperate behavior of mice. Place the C57 mice in a 5L (diameter 11.8 cm, height 27.5 cm) glass beaker (the water level is 18-20 cm high) , water temperature 23-25°C), the mice were visually isolated from each other, and the activities of the mice were recorded for 6 minutes. The time the mice spent immobile in the water within 4 minutes was analyzed by 2 blind persons, where activity was defined as swimming and climbing, and inactivity was defined as floating or making small strokes to keep the head above the water.
  • FST forced swimming test
  • the restraint stress model is another recognized mouse depression model, which can better simulate the "uncontrollable” crowding, frustration and other living conditions in human life. It is similar to the process of human stress and the resulting psychosomatic diseases. , and can reflect the development trend of stress state from acute to chronic, which is in line with the theoretical basis of the onset of depression.
  • the model is highly operable and reproducible, and can well simulate the core symptoms of depression such as behavioral despair and lack of interest.
  • the crocin suspension preparation (0.2 mL/20 g body weight) of the specifications 123 prepared in Example 1 is administered intragastrically, with ketamine as the positive drug (intraperitoneal injection, 10 mg/kg), the control group and the model group was given an equal volume of 0.5% CMC-Na.
  • mice Use the tail suspension test (TST) to evaluate the desperate behavior of mice.
  • TST tail suspension test
  • C57 mice were glued 1 cm from the tip of their tail with medical tape and fixed in a tail suspension box, with their heads about 20 meters above the ground. cm, the mice were visually isolated from each other, and the activities of the mice were recorded on video for 6 minutes.
  • the immobility time of mice in the last 4 minutes was analyzed by 2 blind persons, where immobility was defined as complete stillness or only slight limb movement.
  • Oral dosage 3crocin suspension has a rapid and efficient antidepressant effect on restraint stress model mice, and its efficacy is equivalent to intraperitoneal injection of ketamine.
  • the chronic unpredictable mild stress model is a widely used depression model traditionally. It simulates the chronic low-intensity stress that humans receive in daily life and is close to the disease state of patients with mild to moderate depression. It can simulate the behaviors of depression, such as despair, Core symptoms such as loss of interest.
  • Dosing method After modeling, administer the 30 mg/mL crocin suspension preparation of specification 3 (0.2 mL/20 g body weight) prepared in Example 1 by gavage, with ketamine as the positive drug (intraperitoneal injection, 10 mg/kg), The control group and model group were given equal volumes of 0.5% CMC-Na.
  • mice Use the sugar water preference test (SPT), give 1% sucrose water or drinking water respectively, pre-adapt for 24 hours before the experiment, and change the positions of the sugar water and drinking water bottles every 12 hours, and weigh and record during the experiment The mice drank the weight of sugar water and drinking water respectively within 12 hours at night.
  • the formula for calculating the sugar water preference coefficient is: the weight of drinking sugar water/(the weight of drinking sugar water + the weight of drinking purified water)*100%.
  • the results showed that 24 hours after administration, oral dosage 3 of crocin suspension and intraperitoneal injection of ketamine significantly improved the sugar water preference behavior of mice (see Figure 6A).
  • Oral dosage 3 of crocin suspension has a rapid and efficient antidepressant effect on chronic unpredictable stress model mice, and its effect is equivalent to intraperitoneal injection of ketamine.
  • a social defeat stress model was established in the same manner as in Example 6, and depression-sensitive mice were screened out using the social coefficient, and the experiments were conducted after random grouping.
  • the nucleus of the solitary tract is the site where the vagus nerve enters the brain, and c-fos is a marker of neuronal activation.
  • the brain tissue of mice in each group was taken for c-fos immunofluorescence staining.
  • the results showed that only the crocin mixed with specification 3
  • the suspension group had a large amount of c-fos expression in the nucleus of the solitary tract (see Figure 7A), suggesting that crocin can activate the vagus nerve after accumulation in the intestine.
  • the cAMP levels in the nucleus accumbens were measured.
  • the results showed that the cAMP levels in the nucleus accumbens of depression model mice were significantly lower than those in the control group.
  • Crocin suspension of specification 3 could increase the cAMP levels in the nucleus accumbens (see Figure 7B).
  • crocin suspension can activate the vagus nerve and increase the level of cAMP in the nucleus accumbens after intestinal accumulation.
  • a social defeat stress model was established as in Example 6, and depression-sensitive mice were screened out using social coefficients. After random grouping, they underwent vagotomy or sham surgery. In the vagotomy group, after the mice were anesthetized, the abdomen was depilated and disinfected. The abdominal skin was cut with sterile surgical scissors to find the entrance to the stomach of the mouse. The end of the esophagus connected to the cardia was separated and carefully separated and cut with fine surgical forceps. A vagus nerve was removed from each side, and the abdominal skin was sutured and sterilized. Continuous intraperitoneal injection of antibiotics was given for two days after surgery to prevent infection. Except for not cutting the vagus nerve, the other operations in the sham operation group were the same as those in the operation group.
  • the crocin suspension of specification 3 (0.2 mL/20 g body weight) prepared in Example 1 was administered intragastrically. 24 hours after administration, social avoidance behavior, tail-suspending despair behavior and sugar water preference loss of interest behavior were evaluated. The results showed that social The frustration model plus sham operation group was given crocin, which significantly increased the social coefficient and improved social avoidance behavior. However, the social frustration model plus vagus nerve cutting group was given crocin and there was no significant improvement in social avoidance behavior (see Figure 8A). Similarly, administration of crocin to the sham operation group significantly reduced tail suspension immobility time, while vagotomy offset the antidepressant effect of crocin (see Figure 8B).
  • the sugar water preference experiment showed that administration of crocin to the sham operation group increased the sugar water preference coefficient, while vagus nerve severing further reduced the sugar water preference coefficient of mice, and vagus nerve severing offset most of the improvement effect of crocin (see Figure 8C).
  • nucleus accumbens cAMP levels in the nucleus accumbens brain area were measured. The results showed that the administration of crocin to the sham operation group could increase the cAMP level in the nucleus accumbens brain area. Cutting off the vagus nerve blocked the regulatory effect of crocin on nucleus accumbens cAMP (see Figure 8D ). Comprehensive previous studies have found that nucleus accumbens cAMP is a target of depression regulation, suggesting that the main mechanism for crocin to exert rapid antidepressant effects is to regulate the level of nucleus accumbens cAMP through the vagus nerve.
  • Crocin exerts a rapid antidepressant effect by regulating cAMP in the nucleus accumbens through the vagus nerve.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Inorganic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Neurology (AREA)
  • Dispersion Chemistry (AREA)
  • Nutrition Science (AREA)
  • Pain & Pain Management (AREA)
  • Psychiatry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Physiology (AREA)
  • Neurosurgery (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicinal Preparation (AREA)

Abstract

发明公开了一种藏红花素混悬剂，含有下述质量比的活性成分与辅料：藏红花素：助悬剂=0.2-20:0.4-2，余量为水，其中助悬剂与混悬剂的质量百分比为0.2-0.5%，及其在制备快速抗抑郁药物中的应用。

Description

一种藏红花素混悬剂及其在制备快速抗抑郁药物中的应用 技术领域
本发明属于药物领域，涉及藏红花素混悬剂及其在制备快速抗抑郁药物中的应用，特别涉及藏红花素混悬剂通过迷走神经调控伏隔核脑区环磷酸腺苷(cAMP)水平，发挥快速、安全、高效抗抑郁作用及其应用。
背景技术
抑郁症发病率高，抑郁治疗药物市场空间巨大：抑郁症已成为仅次于癌症的人类第二大杀手。全球抑郁症平均发病率约4.4％，且在近10年来，患病人数增长了18％以上。据2019年北京大学第六医院黄悦勤教授等在《柳叶刀精神病学》上发表文章显示，我国抑郁症的终身患病率为6.9％，12个月患病率为3.6％，据此推算我国抑郁症患者接近1亿人。但我国抑郁症患者就诊率仅为9.5％，在就诊的患者群体中，得到充分治疗的仅有0.5％。近年来，国家政策支持要普遍开展抑郁精神障碍的防治，抑郁症治疗率要在现有基础上提高50％。因此，在刚性需求和政策支持下，抗抑郁药物研发及抗抑郁药物的市场价值上拥有巨大空间。
临床亟需快速、有效、安全治疗药物：据统计，2019年我国抗抑郁药市场销售额为94亿元，全球市场规模达809亿元。然而，现有的一线治疗药物——单胺再摄取抑制剂存在严重弊端：1)通常用药3-4周以上才起效；2)对约1/3抑郁患者无效，且用药人群中约一半患者在治疗后出现复发；3)副作用多，甚至诱发患者自杀。由于相当大比例的抑郁患者存在自杀意念，单胺再摄取抑制剂起效迟缓，在用药(前3-4周)不起效情况下，可能加深患者的失望/绝望情绪，并进一步诱发自杀。因此研发快速起效的抗抑郁药，将起效时间由大约3-4周缩短至一周内甚至1-2天，具有极其重要的临床意义。
由于极度缺乏快速起效药物，有研究发现氯胺酮(属于毒品范畴)单次注射后可在3-4小时缓解抑郁症状，尽管氯胺酮抗抑郁的作用机制仍未明确、有较强的成瘾性，无法大规模使用，但由于确切的抗抑郁作用，美国和欧盟已经相继批准上市了艾斯氯胺酮用于治疗“难治性抑郁症”。此外，在抗抑郁药物研发领域，目前国际上十分关注另外一种快速抗抑郁药裸盖菇素(来自迷幻蘑菇，致幻剂，属于毒品范畴)，研究发现单剂量裸盖菇素可显著减少被试者的抑郁和焦虑。上述两种药物虽能快速缓解抑郁症状，但由于成瘾、致幻等严重毒副 作用，临床上难以广泛应用。因此，亟需快速、有效、安全的抗抑郁药。开发具有明确抗抑郁药效基础上，有着自己独特药效特点和新型抗抑郁作用机制，同时具备副作用相对较小、多靶点、多功效等优点的新型抗抑郁药物将会有广阔的市场前景。
调控伏隔核cAMP可快速、有效治疗抑郁：我们前期研究发现伏隔核中环磷酸腺苷(cAMP)水平对抑郁敏感性至关重要，并证实下调cAMP引起抑郁敏感性增加，上调cAMP水平可发挥抗抑郁作用，提示伏隔核cAMP是抑郁调控重要靶标。然而开发调控伏隔核cAMP的药物面临需透过血脑屏障和特异性作用于伏隔核脑区的两个难题。进一步地，我们经过大量筛选研究，发现口服藏红花素可以调节伏隔核cAMP水平，并且显著改善实验动物的抑郁样行为。静脉注射藏红花素未见明显药效，并且切断肠道与中枢之间联通的迷走神经后，药效消失。证明口服藏红花素的药效必定依赖于其进入肠道。进一步研究还发现较高剂量藏红花素经口给药方式可在盲肠和结肠部位蓄积，在肠道通过激活迷走神经、经神经回路调控提高伏隔核脑区cAMP水平，发挥快速抗抑郁作用，而较低剂量下，盲肠和结肠部位药物量很少，短时间内难以出现明显药效(需要二周左右才能起效)。这表明藏红花素具有通过肠道发挥快速抗抑郁作用的机制，不仅规避了药物需要通过血脑屏障入脑的难题，又借助机体自身的神经回路调控特异性作用于伏隔核脑区，还具有快速起效和安全性好的药效特点，且口服给药患者依从性较好。将藏红花素制备成有利于其在肠道发挥快速抗抑郁作用的制剂，口服给药后，能够在1-2天内快速缓解患者的抑郁症状，具有极其重要的临床意义和广阔的市场前景。
然而，藏红花素在肠道发挥快速抗抑郁作用缺乏有效的制剂：藏红花素在普通生理环境下溶解度不高，生物膜透过能力较弱，在偏酸、偏碱环境中不稳定，容易降解成为其它物质而降低药效。由于普通片剂、胶囊剂口服后进入胃内，在胃中逐渐崩解，藏红花素与辅料会较长时间滞留于胃内皱褶、粘附在胃壁，造成在胃酸性环境中受到破坏，直接减少进入肠道后发挥药效的药物量，不利于药效的有效发挥。
发明内容
本发明的目的是针对现有抑郁治疗药物的缺乏和技术的上述不足，提供藏红花素在制备混悬剂及其在快速抗抑郁药物中的应用。
本发明的目的可通过如下技术方案实现：
一种藏红花素混悬剂，其特征在于含有下述质量比的活性成分与辅料：藏红花素：助悬剂比例为0.2-20:0.4-2，余量为水，其中助悬剂与混悬剂的质量百分比为0.2-0.5％。
我们经过近十年的研究发现藏红花素通过在肠道激活迷走神经，并经肠脑神经回路调控伏隔核cAMP水平发挥快速抗抑郁作用的新机制，且藏红花素通过这种机制发挥作用需要在盲肠和结肠部位大量且长时间蓄积。因此分别制备藏红花素混悬剂、颗粒剂和溶液剂，对藏红花素在肠道蓄积情况进行考察，筛选得到藏红花素混悬剂为优选制剂。对助悬剂种类羧甲基纤维素钠(CMC-Na)和羟丙基甲基纤维素(HPMC)对藏红花素在肠道蓄积的影响进行考察，筛选得到CMC-Na为优选助悬剂。进一步对CMC-Na的浓度对藏红花素在肠道蓄积的影响进行考察，筛选得到0.2％-0.5％CMC-Na更有助于藏红花素在肠道蓄积。考虑到藏红花素须在肠道蓄积以便充分发挥药效，发明人研究发现当藏红花素制成混悬剂形式，能够在肠道蓄积和持续产生作用，获得非常好的治疗效果。
本发明的所述的藏红花素结构如下：
分子式：C44H64O24；分子量：976.96。
本发明所述助悬剂选自艾维素CL611、艾维素CL591、羟丙基纤维素、羟乙基纤维素、羧甲基纤维素钠、乙基纤维素、甲基纤维素、黄原胶、海藻酸钠、聚乙烯醇、聚维酮k-30、聚维酮k-90、聚乙烯吡咯烷酮。
作为本发明的一种优选，所述的助悬剂为羧甲基纤维素钠。
作为本发明的一种优选，每单位混悬剂的藏红花素：羧甲基纤维素钠的质量比为3-20:0.4-1。
作为本发明的更进一步优选，每单位混悬剂的藏红花素：羧甲基纤维素钠的质量比为4-8:1。
作为本发明的更进一步优选，每单位混悬剂的藏红花素：羧甲基纤维素钠的质量比为6:1。
作为本发明的一种优选，所述的藏红花素混悬剂是将活性成分藏红花素与所述的助悬剂经过简单物理混合，再溶于水中成为混悬剂；或者将活性成分藏红花素分散于事先配制好的助悬剂水溶液中，成为混悬剂。
作为本发明的一种优选，每制剂含有藏红花素浓度为1-100mg/mL，优选15-30mg/mL。
本发明所述的藏红花素混悬剂在制备快速起效的抗抑郁药物中的应用，所述的快速起效为24-48小时内起效。
作为本发明的一种优选，藏红花素混悬剂人体给药剂量为以藏红花素计，0.5-4g/60kg，优选1-2g/60kg。
作为本发明的一种优选，所述藏红花素混悬剂中，其中所述的藏红花素的单位剂量为以藏红花素计为50-400mg。作为本发明的一种优选，所述混悬剂中所述的藏红花素的单位剂量为以藏红花素计为100-200mg；前述高剂量藏红花素单次给药，24小时-48h内即可显示明显的快速抗抑郁效果。
作本发为本发明的进一步优选，所述药物制剂的剂型包括口服给药形式的剂型。
作为本发明的一种优选，所述藏红花素口服混悬剂中，其中所述的藏红花素的单位剂量为以藏红花素计为50-400mg。作为本发明的一种优选，所述藏红花素口服混悬剂中所述的藏红花素单位剂量为以藏红花素计为100-200mg；前述藏红花素口服混悬剂单次口服给药，24小时-48h内即可显示明显的快速抗抑郁效果。作为本发明的一种优选，藏红花素混悬剂人体给药体积以混悬剂计，体积为5-50mL,优选10-20mL。
作为本发明的一种优选，藏红花素混悬剂口服给药后，活性成分藏红花素在盲肠和结肠部位显著蓄积时，在肠道通过迷走神经调控升高伏隔核cAMP水平发挥快速抗抑郁作用。
发明人基于发现伏隔核中环磷酸腺苷(cAMP)水平对抑郁敏感性至关重要，并证实下调cAMP引起抑郁敏感性增加，上调cAMP水平可发挥抗抑郁作用，提示伏隔核cAMP是抑郁调控重要靶标，经过大量筛选和研究发现藏红花素混悬剂口服给药后在肠道蓄积，并通过迷走神经调控升高伏隔核cAMP水平发挥快速抗抑郁作用，切断迷走神经阻断藏红花素升高伏隔核cAMP并抵消其抗抑郁作用。重要的是，由国内权威实验室使用两个经典的抑郁模型在双盲的情况下均证实了高剂量藏红花素混悬剂可在给药后24-48小时快速改善抑郁症状。证明藏红花素通过全新的调控机制发挥快速、高效抗抑郁作用，可用于快速抗抑郁药物的制备。
本发明所述藏红花素来源包括但不限于通过茜草科植物大花栀子、栀子的果实、鸢尾科植物藏红花及同属其它植物的花柱头等、马钱科植物密蒙花、木犀科植物夜花、菊科植物牛蒡、百部科植物蔓生百部都豆科植物含羞草等为原料，利用现代的生物技术提取，以及其它人工合成的藏红花素。
有益效果：
藏红花是传统名贵中药，无毒，用于心忧郁积、气闷不散、活血，久食令人心喜。我们研究发现中药藏红花中的藏红花素可通过激活迷走神经提高伏隔核脑区cAMP水平，并在给药24-48小时快速改善抑郁症状，其口服药效强度与腹腔注射的氯胺酮相当。重要的是，藏红花素的快速抗抑郁作用在三种公认的抑郁模型、三个独立实验室中在双盲的情况下都得到验证。另外研究发现静注相当量的藏红花素无效，藏红花素混悬剂口服给药后在肠道蓄积时，通过迷走神经调控升高伏隔核cAMP水平发挥快速抗抑郁作用，切断迷走神经阻断藏红花素升高伏隔核cAMP并抵消其抗抑郁作用。这表明藏红花素是一种新型、快速的抗抑郁药物，具有独特的通过肠道迷走神经、经神经回路调控伏隔核cAMP发挥快速抗抑郁作用，具有起效快和安全性好的药效特点，且口服给药患者依从性较好，藏红花素混悬剂能实现足量的藏红花素在肠道的有效蓄积，从而通过全新机制发挥抗抑郁药效。
本发明所述工艺制备的藏红花素混悬剂有利于减少在胃酸环境下的降解，藏红花素混悬剂是具有一定粘度的混悬液状态，口服后液体不但会比固体颗粒更快从胃排空进入肠道，而且混悬剂中的颗粒大部分分散于具有一定粘度的溶液中，减少了与酸性胃液的接触和在胃中的直接暴露。上述二重因素也导致混悬液中藏红花素更少受到胃液的影响和破坏，增加进入肠道的药物量，更有利于在肠道中发挥药效。
本发明明确了藏红花素通过全新机制发挥抗抑郁药效，填补了国际上快速、安全、高效抗抑郁治疗药物的空白，将有力支持其作为抗抑郁药物的研发，惠及全球数以亿计抑郁症患者，产生巨大经济效益和深远社会效益。
附图说明
图1三种藏红花素制剂给药后2h、6h和12h在各肠段肠内容物中藏红花素浓度统计图。
图2不同助悬剂制得的藏红花素混悬剂灌胃给药后2h、6h和12h在各肠段肠内容物中藏红花素浓度统计图。
其中，A：以羧甲基纤维素钠(CMC-Na)为助悬剂制得的藏红花素混悬剂；B：以羟丙基甲基纤维素(HPMC)为助悬剂制得的藏红花素混悬剂；①、②、③分别为以CMC-Na为助悬剂制得的规格7.5、15、30mg/mL的藏红花素混悬剂，④、⑤、⑥分别为以HPMC为助悬剂制得的规格7.5、15、30mg/mL的藏红花素混悬剂。
图3不同CMC-Na浓度制得的藏红花素混悬剂给药后2h、6h和12h在各肠段肠内容物中藏红花素浓度统计图。
图4藏红花素混悬剂在社交挫败应激模型中快速抗抑郁。
其中，A：各组小鼠社交系数统计图；B：各组小鼠强迫游泳不动时间统计图；①、②、③分别为以CMC-Na为助悬剂制得的规格7.5、15、30mg/mL的藏红花素混悬剂(0.2mL/g体重)，氯胺酮给药剂量为10mg/kg(腹腔注射)，采用one-way ANOVA分析，*p＜0.05，**p＜0.01，***p＜0.001。
图5藏红花素混悬剂在束缚应激模型中快速抗抑郁。
其中，A：各组小鼠悬尾不动时间统计图；B：各组小鼠强迫游泳不动时间统计图；①、②、③分别为以CMC-Na为助悬剂制得的规格7.5、15、30mg/mL的藏红花素混悬剂(0.2mL/g体重)，氯胺酮给药剂量为10mg/kg(腹腔注射)，采用one-way ANOVA分析，*p＜0.05，**p＜0.01。
图6高剂量藏红花素混悬剂在慢性不可预知应激模型中快速抗抑郁。
其中，A：各组小鼠糖水偏好系数统计图；B：各组小鼠强迫游泳不动时间统计图；③是以CMC-Na为助悬剂制得的规格30mg/mL的藏红花素混悬剂(0.2mL/g体重)，氯胺酮给药剂量为10mg/kg(腹腔注射)，采用one-way ANOVA分析，***p＜0.001，****p＜0.0001。
图7高剂量藏红花素混悬剂在肠道蓄积并激活迷走神经。
其中，A：各组小鼠孤束核(NTS)c-fos表达情况免疫荧光染色图；B：各组小鼠伏隔核cAMP水平统计图；①、②、③分别为以CMC-Na为助悬剂制得的规格7.5、15、30mg/mL的藏红花素混悬剂(0.2mL/g体重)，氯胺酮给药剂量为10mg/kg(腹腔注射)，采用one-way ANOVA分析，*p＜0.05，**p＜0.01。
图8切断迷走神经抵消藏红花素的抗抑郁作用。
其中，A：各组小鼠社交系数统计图；B：各组小鼠悬尾不动时间统计图；C：各组小鼠糖水偏好系数统计图；D：各组小鼠伏隔核脑区cAMP测定浓度统计图；空白溶剂表示灌胃给予等体积0.5％CMC-Na，采用two-way ANOVA分析，*p＜0.05，**p＜0.01，***p＜0.001，****p＜0.0001，n.s.表示无统计差异。
具体实施方式
以下通过具体实施例的方式，对本发明做进一步详述，但不应理解为是对本发明的限制。本领域普通技术人员根据上述技术方案，还可以做出多种形式的调整、修改、替换、变更、 改进。凡基于上述技术思想所作的调整、修改、替换、变更、改进均属于本发明的范围。
本发明通过下面的实施例进行详细的解释，但并不意味着本发明仅限于此。
实施例1 藏红花素颗粒剂、溶液剂和混悬剂的制备
(1)藏红花素颗粒剂制备工艺：
制备工艺：取藏红花素5.0g，所述藏红花素(由藏红花或栀子中提取，购自曼思特生物科技公司，纯度99.07％，批号MUST-20070213)过60目筛；乳糖(乳糖，德国美剂乐MEGGLE，批号101504319)2.0g过80目筛；微晶纤维素(德国JRS，批号5610264626)3.0g，过80目筛。将藏红花素与乳糖、微晶纤维素充分混合，加入适量0.5％羧甲基纤维素钠(国药集团，批号20131022)制成软材，再使用20目筛分散成颗粒，烘干，每制剂0.5g。
(2)藏红花素溶液剂制备工艺：
以PEG400作为助溶剂，规格30mg/mL
取藏红花素3.0g，过60目筛，加入50mL 90℃的热水溶解，加入30mL PEG400(上海凌峰，批号20200608)混匀，再加入20mL热水，一共100mL，混匀后分装，每制剂10mL，新鲜配制后给药。
(3)藏红花素混悬剂制备工艺：
以羧甲基纤维素钠(CMC-Na)为助悬剂：(规格①、②、③分别为7.5、15、30mg/mL)
藏红花素        0.75g/1.5g/3.0g
羧甲基纤维素钠  0.5g
蒸馏水          加至100mL
制备工艺：分别取藏红花素0.75g/1.5g/3.0g，过60目筛；羧甲基纤维素钠(国药集团，批号20131022)0.5g过80目筛。将羧甲基纤维素钠分散并溶于100mL蒸馏水中，临用时再将藏红花素分别分散混悬于0.5％羧甲基纤维素钠水溶液中，震荡混悬均匀，每制剂10mL，新 鲜配制后给药。
以羟丙基甲基纤维素(HPMC)为助悬剂：(规格④、⑤、⑥分别为7.5、15、30mg/mL)
藏红花素          0.75g/1.5g/3.0g
羟丙基甲基纤维素  0.5g
蒸馏水            加至100mL
制备工艺：分别取藏红花素0.75g/1.5g/3.0g，过60目筛；羟丙基甲基纤维素(安徽山河药用辅料公司，批号181231)0.5g过80目筛。将羟丙基甲基纤维素分散并溶于100mL蒸馏水中，临用时再将藏红花素分别分散混悬于0.5％羟丙基甲基纤维素水溶液中，震荡混悬均匀，每制剂10mL，新鲜配制后给药。
以不同浓度的羧甲基纤维素钠(CMC-Na)为助悬剂：(规格⑦、⑧分别为0.2％、1.0％CMC-Na)
藏红花素          3.0g
羧甲基纤维素钠    0.2g/1.0g
蒸馏水            加至100mL
制备工艺：分别取藏红花素3.0g，过60目筛；分别取羧甲基纤维素钠0.2g/1.0g过80目筛。分别将羧甲基纤维素钠分散并溶于100mL蒸馏水中，临用时再将藏红花素分别分散混悬于0.2％/1.0％羧甲基纤维素钠水溶液中，震荡混悬均匀，每制剂10mL，新鲜配制后给药。
以不同量藏红花素制备混悬剂，以羧甲基纤维素钠(CMC-Na)为助悬剂：(规格⑨、⑩分别为1.0、100mg/mL)
藏红花素          0.1g/10.0g
羧甲基纤维素钠    0.5g
蒸馏水            加至100mL
制备工艺：分别取藏红花素0.1g/10g，过60目筛；羧甲基纤维素钠0.5g过80目筛。将羧甲基纤维素钠分散并溶于100mL蒸馏水中，临用时再将藏红花素分别分散混悬于0.5％羧甲基纤维素钠水溶液中，震荡混悬均匀，每制剂10mL，新鲜配制后给药。
实施例2 三种剂型的藏红花素给药后在肠道蓄积情况
选择C57BL/6J小鼠(7-8周)，每组6只，分别灌胃给予实施例1制备的藏红花素颗粒剂、藏红花素溶液剂和藏红花素混悬剂(规格③)(以藏红花素计，给药剂量为300mg/kg)，考察不同藏红花素制剂灌胃给药后在肠道蓄积情况。在给药后2h、6h和12h分别取空肠、回肠、盲肠和结肠内容物，以100mg肠道内容物/1mL水的比例加超纯水充分混匀后，离心取上清测定藏红花素的浓度。结果显示，在给药后2h，给予颗粒剂组藏红花素在回肠中浓度更高，但给药后6h-12h，混悬剂组藏红花素在盲肠和结肠中浓度更高(参见图1)。在小鼠使用藏红花素的常规药理剂量50-300mg/kg范围内，较高剂量时使用藏红花素混悬剂更有利于藏红花素在盲肠和结肠内容物中长时间蓄积。
结论：藏红花素混悬剂剂型更有利于藏红花素在盲肠和结肠中长时间蓄积。
实施例3 三种剂型的理化性质
(1)取本发明实施例1制备的三种藏红花素制剂，观察在室温存放前后外观性状变化。结果显示藏红花素混悬剂和颗粒剂较为稳定，存放一周后未见明显性状变化，而溶液剂存放一周后出现明显分层，参见表1。
表1三种剂型在室温放置1周前后变化
实施例4 助悬剂种类和浓度筛选
为筛选更有效的助悬剂种类，选择C57BL/6J小鼠(7～8周)，每组5只，分别灌胃给予实施例1制备的规格①②③④⑤⑥的藏红花素混悬剂(0.2mL/20g体重)，考察不同藏红花素混悬剂灌胃给药后在肠道蓄积情况。在给药后2h、6h和12h分别取空肠、回肠、盲肠和结肠内容物，以100mg/1mL的比例加超纯水充分混匀后，离心取上清测定藏红花素浓度。结果显示，在小鼠使用藏红花素的常规药理剂量50-300mg/kg范围内，相比羟丙基甲基纤维素(HPMC)，以羧甲基纤维素钠(CMC-Na)为助悬剂制得的藏红花素混悬剂，在给药后6h至12h，藏红花素在盲肠和结肠中浓度更高，且停留时间更长(参见图2)。
为筛选更优的助悬剂CMC-Na的浓度，选择C57BL/6J小鼠(7-8周)，每组5只，分别灌胃给 予实施例1制备的规格③⑦⑧的藏红花素混悬剂(0.2mL/20g体重)，考察不同藏红花素制剂灌胃给药后在肠道蓄积情况。在给药后2h、6h和12h分别取空肠、回肠、盲肠和结肠内容物，以100mg/1mL的比例加超纯水充分混匀后，离心取上清测定藏红花素浓度。结果显示，相比以1.0％CMC-Na为助悬剂，使用0.2％或0.5％CMC-Na为助悬剂制得的藏红花素混悬剂，在给药后2h、6h和12h，藏红花素在各肠段中的浓度更高(参见图3)。
本发明还选择了其它助悬剂进行试验，所述助悬剂包括但不限于(1)低分子助悬剂，如甘油、糖浆等；(2)高分子助悬剂①天然：阿拉伯胶、西黄蓍胶、琼脂、海藻酸钠、白及胶、果胶等；②合成类如艾维素CL611、艾维素CL591、羟丙基纤维素、羟乙基纤维素、乙基纤维素、甲基纤维素、黄原胶、海藻酸钠、聚乙烯醇、聚维酮k-30、聚维酮k-90、聚乙烯吡咯烷酮等；③硅酸类如胶体二氧化硅、硅酸铝、硅皂土。试验结果发现羧甲基纤维素钠为最优选的助悬剂。
实施例5 藏红花素混悬剂的急性毒性试验
供试品：取上述规格③的藏红花素混悬剂，含30mg/mL藏红花素。
检测方法：取体重18～22g小鼠20只，每组10只，分别灌胃给予0.2mL混悬液，以藏红花素计，相当于给药剂量300mg/kg，连续给药7天，小鼠未见死亡或异常。
实施例6 藏红花素混悬剂对社交挫败应激模型小鼠的快速抗抑郁作用
慢性社交挫败应激模型是近年来最为广泛使用的抑郁小鼠模型，这种模型因具有明显的诱因(社交挫败应激)、建模成功率高、具有明显的表型呈现而在专业领域被广泛认可。由于大多数抑郁相关的应激具有社会性特征，包括环境压力、生活压力事件和人际交往挫折等，其本质上是社会性的，慢性社交挫败应激模型能更好地模拟社会性因素导致抑郁的真实过程，与人类抑郁症特征具有很高的相似性，此外还具有持久的行为学和生物学变化，是研究抑郁症最好的动物模型之一。
慢性社交挫败模型采用CD-1小鼠(退役鼠，雄性，4-6个月，购自北京维通利华实验动物技术有限公司，体重48±5g)和C57BL/6J小鼠(雄性，7～8周，购自北京维通利华实验动物技术有限公司，体重20±2g)，在标准饲养环境下(自由饮食与饮水，昼夜交替，各12小时)，适应性饲养一周。
动物模型的建立：对CD-1小鼠连续筛选3天，筛选出有攻击性的CD-1小鼠用于造模。将CD-1小鼠和C57小鼠用带孔透明塑料隔板隔开，每天将C57小鼠放置在CD-1小鼠同侧使其接受攻击， 每次10分钟，每天接受一只新的CD-1小鼠的攻击，结束后将C57小鼠放在对侧，在剩余24小时内继续在视觉和味觉上接受CD-1小鼠的应激。连续造模10天。在第11天根据社交回避行为筛选社交系数小于1的小鼠(即抑郁敏感鼠)进行试验。
给药方式：对抑郁敏感鼠灌胃给予实施例1制备的规格①②③的藏红花素混悬制剂(0.2mL/20g体重)，以氯胺酮为阳性药(腹腔注射，10mg/kg)，对照组和模型组给予等体积0.5％CMC-Na。
对小鼠社交回避行为的影响：给药后24小时进行社交行为评价。实验前将小鼠置于行为学测试房间适应光线和温度30分钟以上。将C57小鼠放于一个40×40×40cm的旷场内，同时在社交区域放入一个空的网笼，记录C57小鼠在2.5分钟内的活动情况，结束后将C57小鼠移走并清理旷场区域。第二段时间在网笼内放入一只新的CD-1小鼠，同样记录2.5分钟。社交系数(SI)计算方法为C57小鼠在第二段时间内进入社交区域的时间与第一段时间内进入社交区域的时间的比值。
结果表明，模型组小鼠的社交系数与对照组相比显著下降，规格③藏红花素和阳性药氯胺酮可显著提高社交系数，其他组的藏红花素对社交回避行为没有明显改善作用(参见图4A)。
考虑到氯胺酮是目前唯一一个已上市的高效、快速抗抑郁药物(数小时内起效)，本专利工艺制备的藏红花素混悬剂口服后具有接近氯胺酮的抗抑郁效果，而且无明显毒副作用，有望成为新一代高效、快速、低毒抗抑郁药。
对小鼠绝望行为的影响：使用强迫游泳实验(FST)评价小鼠的绝望行为，将C57小鼠放于一个5L(直径11.8厘米，高27.5厘米)的玻璃烧杯中(水位高18～20厘米，水温23-25℃)，小鼠彼此之间视觉隔离，记录小鼠活动情况，持续6分钟。由2名单盲人员分析后4分钟内小鼠在水中静止不动的时间，其中活动定义为游泳和攀爬，静止定义为漂浮不动或为了保持头部抬出水面而小幅度划水。
结果表明，抑郁模型小鼠的不动时间与对照组相比显著增加，提示小鼠出现绝望行为，口服规格③藏红花素混悬剂和腹腔注射氯胺酮显著降低不动时间，改善了小鼠的绝望行为(参见图4B)。
进一步地，时程效应研究发现规格③的藏红花素混悬剂给药后在2h开始显现改善抑郁行为的作用，在给药后24h显著改善抑郁样行为，作用持续48h。安全性评价研究结果显示本发明所用的藏红花素给药剂量不引起肝肾损伤，对各种神经和精神行为无明显影响。
结论：口服规格③藏红花素对社交挫败应激模型小鼠具有快速、高效抗抑郁作用，其药 效与腹腔注射的氯胺酮相当。同时，口服藏红花素安全性好，不会引起成瘾问题。
实施例7 藏红花素对束缚应激模型小鼠的快速抗抑郁作用
束缚应激模型是另一种公认的小鼠抑郁模型，能够较好地模拟人类生活中“无法控制”的拥挤、挫折等生活状态，既与人类产生应激及其导致的心身性疾病过程相似，又能够体现应激状态由急性转入慢性的发展趋势，符合抑郁症发病的理论基础。该模型可操作性强，重复性好，并能够很好模拟抑郁症的行为绝望、兴趣缺失等核心症状。
动物模型的建立：同实施例6选择C57小鼠，将50mL EP管打孔用于束缚应激，造模时每天在固定时间将C57小鼠放入带孔的50mL EP管，并注意在每只小鼠之间放置隔板，每天4-6小时，连续21天。
给药方式：造模结束后灌胃给予实施例1制备的规格①②③的藏红花素混悬制剂(0.2mL/20g体重)，以氯胺酮为阳性药(腹腔注射，10mg/kg)，对照组和模型组给予等体积0.5％CMC-Na。
对小鼠绝望行为的影响：使用悬尾实验(TST)评价小鼠绝望行为，将C57小鼠分别用医用胶带粘住尾尖1厘米处并固定于悬尾箱中，头部离地约20厘米，小鼠彼此之间视觉隔离，录像记录小鼠的活动情况，持续记录6分钟。由2名单盲人员分析小鼠在后4分钟的不动时间，其中不动定义为完全静止不动或只有轻微的四肢运动。
结果表明，束缚应激造模显著增加小鼠不动时间，口服规格③藏红花素混悬剂和腹腔注射氯胺酮显著改善小鼠的绝望行为(参见图5A)。
另外同实施例6使用强迫游泳实验评价小鼠的绝望行为，结果表明，模型组小鼠强迫游泳不动时间显著增加，规格③藏红花素混悬剂和氯胺酮显著降低不动时间(参见图5B)。
结论：口服规格③藏红花素混悬剂对束缚应激模型小鼠具有快速、高效抗抑郁作用，其药效与腹腔注射的氯胺酮相当。
实施例8 藏红花素对慢性不可预知应激模型小鼠的快速抗抑郁作用
慢性不可预知温和应激模型是传统上广泛使用的一种抑郁模型，模拟人类日常生活中接受的慢性低强度应激，接近轻中度抑郁症患者的疾病状态，能够模拟抑郁症的行为绝望、兴趣缺失等核心症状。
动物模型的建立：同实施例6选择C57小鼠，将小鼠用1％蔗糖水适应后，连续2周测定糖水偏好系数作为基础值。第2周结束后，根据糖水偏好系数随机分组。每天随机给予2种以下 应激：冰水游泳(10分钟)，束缚(4小时)，潮湿垫料(12小时)，禁水禁食(24小时)，社交挫败(10分钟)，悬尾(30分钟)，夜间光照(12小时)，连续4周。
给药方式：造模结束后灌胃给予实施例1制备的规格③的30mg/mL的藏红花素混悬制剂(0.2mL/20g体重)，以氯胺酮为阳性药(腹腔注射，10mg/kg)，对照组和模型组给予等体积0.5％CMC-Na。
对小鼠兴趣丧失行为的影响：选用糖水偏好实验(SPT)，分别给予1％蔗糖水或饮用水，实验前预适应24h，且每12h调换糖水和饮用水的水瓶位置，实验时称重记录夜间12h内小鼠分别饮用糖水和饮用水的重量，糖水偏好系数计算公式为：饮用糖水重量/(饮用糖水重量+饮用纯净水重量)*100％。结果表明，给药后24小时，口服规格③的藏红花素混悬剂和腹腔注射的氯胺酮显著提高小鼠的糖水偏好行为(参见图6A)。
另外，同实施例6使用强迫游泳实验评价小鼠的绝望行为，结果表明，在慢性不可预知应激模型中，模型组小鼠的不动时间显著增加，单次灌胃给予规格③的藏红花素混悬剂和腹腔注射氯胺酮后24小时显著降低不动时间(参见图6B)。
结论：口服规格③的藏红花素混悬剂对慢性不可预知应激模型小鼠具有快速、高效抗抑郁作用，其作用与腹腔注射氯胺酮相当。
实施例9 藏红花素混悬剂对迷走神经的作用
动物模型的建立：同实施例6建立社交挫败应激模型，使用社交系数筛选出抑郁敏感小鼠，随机分组后进行实验。
孤束核(NTS)是迷走神经入脑的部位，c-fos是神经元激活的标志物，取各组小鼠脑组织进行c-fos免疫荧光染色，结果显示，仅有规格③的藏红花素混悬剂组在孤束核有大量c-fos表达(参见图7A)，提示藏红花素在肠道蓄积后可激活迷走神经。测定伏隔核cAMP水平，结果显示，抑郁模型小鼠伏隔核cAMP水平相比对照组显著下降，规格③的藏红花素混悬剂可升高伏隔核cAMP水平(参见图7B)。
体内分布研究表明只有规格③的藏红花素混悬剂口服后在盲肠和结肠部位显著蓄积，规格①②没有明显蓄积。因此，藏红花素混悬剂在肠道蓄积后可激活迷走神经并升高伏隔核cAMP水平。
结论：藏红花素混悬剂口服后在盲肠和结肠部位蓄积时，可在肠道通过激活迷走神经提高伏隔核cAMP水平产生抗抑郁作用。
实施例10 切断迷走神经抵消藏红花素的抗抑郁作用
动物模型的建立：同实施例6建立社交挫败应激模型，使用社交系数筛选出抑郁敏感小鼠，随机分组后，做迷走神经切断或假手术。迷走神经切断组，在小鼠麻醉后，将腹部脱毛处理并消毒，用灭菌手术剪刀切开腹部皮肤，找到小鼠胃入口处，分离与贲门相连的食道末端，用细手术镊小心分离并剪断两侧各一条迷走神经，缝合腹部皮肤并消毒。术后两天连续腹腔注射抗生素防止感染。假手术组除不剪断迷走神经外，其余操作与手术组相同。
灌胃给予实施例1制备的规格③的藏红花素混悬剂(0.2mL/20g体重)，给药24小时后，评价社交回避行为、悬尾绝望行为和糖水偏好兴趣缺失行为，结果表明，社交挫败模型复合假手术组给予藏红花素后显著提高社交系数，改善了社交回避行为，而社交挫败模型复合切断迷走神经组给予藏红花素后对社交回避行为没有明显改善(参见图8A)。同样，假手术组给予藏红花素可显著降低悬尾不动时间，而迷走神经切断后抵消了藏红花素的抗抑郁作用(参见图8B)。糖水偏好实验表明，假手术组给予藏红花素可提高糖水偏好系数，而切断迷走神经进一步降低了小鼠糖水偏好系数，且切断迷走神经抵消了大部分藏红花素的改善作用(参见图8C)。
此外，测定伏隔核脑区cAMP水平，结果表明，假手术组给予藏红花素可提高伏隔核脑区cAMP水平，切断迷走神经后阻断了藏红花素对伏隔核cAMP的调控作用(参见图8D)。综合前期研究发现伏隔核cAMP为抑郁调控靶标，提示藏红花素发挥快速抗抑郁药效的主要机制为通过迷走神经调控伏隔核cAMP水平。
结论：藏红花素通过迷走神经调控伏隔核cAMP发挥快速抗抑郁作用。
对于本领域技术人员，本公开不只局限于前述说明性实施例，在不脱离其必要属性的情况下能以其它特定形式体现。因此期望认为，所有方面均作为说明性而不是限制性、对所附权利要求进行参考的实施例而不是前述实施例，引用文献只是针对附加的权利要求而不是上述的实例，以及落入权利要求等效性的含义和范围之内的所有变化因此预期包含于此。
本说明书中列举的所有专利、专利申请和文献参考均在此以其全部内容引入作为参考。在不一致的情况下，包括定义的本公开将是有说服力的。

Claims (10)

  1. 一种藏红花素混悬剂，其特征在于含有下述质量比的活性成分与辅料：藏红花素：助悬剂＝0.2-20:0.4-2，余量为水，其中助悬剂与混悬剂的质量百分比为0.2-0.5％。
  2. 根据权利要求1所述的藏红花素混悬剂，其特征在于所述的助悬剂为羧甲基纤维素钠。
  3. 根据权利要求2所述的藏红花素混悬剂，其特征在于所述藏红花素：羧甲基纤维素钠的质量比为3-20:0.4-1。
  4. 根据权利要求3所述的藏红花素混悬剂，其特征在于所述藏红花素：羧甲基纤维素钠的质量比为4-8:1，优选羧甲基纤维素钠的质量比为6:1。
  5. 根据权利要求1～4中任一项所述的藏红花素混悬剂，其特征在于所述的藏红花素混悬剂是将活性成分藏红花素与所述的助悬剂经过简单物理混合，再溶于水中成为混悬剂；或者将活性成分藏红花素分散于事先配制好的助悬剂水溶液中，成为混悬剂。
  6. 根据权利要求5所述的藏红花素混悬剂，其特征在于所述的藏红花素过40-80目筛，助悬剂羧甲基纤维素钠过60-100目筛。
  7. 权利要求1～4中任一项所述的藏红花素混悬剂在制备快速起效的抗抑郁药物中的应用。
  8. 根据权利要求7所述的应用，其特征在于所述的快速起效为24-48小时内起效。
  9. 根据权利要求8所述的应用，其特征在于所述的藏红花素混悬剂口服给药后，活性成分藏红花素在盲肠和结肠部位显著蓄积，在肠道通过迷走神经调控升高伏隔核cAMP水平发挥快速抗抑郁作用。
  10. 根据权利要求7所述的应用，其特征在于所述的藏红花素混悬剂人体给药剂量为以藏红花素计，0.5-4g/60kg，优选1-2g/60kg。
PCT/CN2023/102757 2022-06-06 2023-06-27 一种藏红花素混悬剂及其在制备快速抗抑郁药物中的应用 Ceased WO2023237124A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202210631541.9A CN115105471B (zh) 2022-06-06 2022-06-06 一种藏红花素混悬剂及其在制备快速抗抑郁药物中的应用
CN202210631541.9 2022-06-06

Publications (1)

Publication Number Publication Date
WO2023237124A1 true WO2023237124A1 (zh) 2023-12-14

Family

ID=83325995

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2023/102757 Ceased WO2023237124A1 (zh) 2022-06-06 2023-06-27 一种藏红花素混悬剂及其在制备快速抗抑郁药物中的应用

Country Status (2)

Country Link
CN (1) CN115105471B (zh)
WO (1) WO2023237124A1 (zh)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115105471B (zh) * 2022-06-06 2023-01-20 中国药科大学 一种藏红花素混悬剂及其在制备快速抗抑郁药物中的应用
CN116585332B (zh) * 2022-06-06 2026-02-06 中国药科大学 高剂量藏红花素在制备快速抗抑郁药物中的应用
CN120391392A (zh) * 2025-07-02 2025-08-01 北京大学第六医院 病理性社交记忆泛化小鼠模型的构建方法及其应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101791314A (zh) * 2009-02-04 2010-08-04 复旦大学 藏红花素在制备催眠类药物中的用途
CN110283220A (zh) * 2019-06-11 2019-09-27 浙江工业大学 西红花苷-1的分离纯化方法及其应用
CN113730387A (zh) * 2021-10-18 2021-12-03 中国药科大学 藏红花素在制备修复血脑屏障损伤药物中的应用
CN115105471A (zh) * 2022-06-06 2022-09-27 中国药科大学 一种藏红花素混悬剂及其在制备快速抗抑郁药物中的应用

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107648346A (zh) * 2017-10-26 2018-02-02 暨南大学 藏红花色素类组合物在制备治疗或改善抑郁症的药物中的应用

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101791314A (zh) * 2009-02-04 2010-08-04 复旦大学 藏红花素在制备催眠类药物中的用途
CN110283220A (zh) * 2019-06-11 2019-09-27 浙江工业大学 西红花苷-1的分离纯化方法及其应用
CN113730387A (zh) * 2021-10-18 2021-12-03 中国药科大学 藏红花素在制备修复血脑屏障损伤药物中的应用
CN115105471A (zh) * 2022-06-06 2022-09-27 中国药科大学 一种藏红花素混悬剂及其在制备快速抗抑郁药物中的应用

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CHEN CHUN-LIN; HU LING-RONG; YANG LING; TONG YAN-LING; HU WANG-SHU: "RAPID ANTIDEPRESSANT EFFECT OF CROCIN BASED ON mTOR-BDNF SIGNALING PATHWAY", JOURNAL OF JINGGANGSHAN UNIVERSITY (NATURAL SCIENCE), vol. 43, no. 3, 31 May 2022 (2022-05-31), pages 41 - 46, XP009550982, ISSN: 1674-8085 *
WANG PING, LIN SUSU, CHEN ZHUOYI, JIANG YU, JIANG JINYAN, LIU LING: "Antidepressant Effects of Crocin-1 from Saffron in Mice with Chronic Unpredictable Mild Stress-Induced Depression", THE FASEB JOURNAL, FEDERATION OF AMERICAN SOCIETIES FOR EXPERIMENTAL BIOLOGY, UNITED STATES, vol. 33, no. s1, 1 April 2019 (2019-04-01), United States , pages lb75, XP009550981, ISSN: 1530-6860, DOI: 10.1096/fasebj.2019.33.1_supplement.lb75 *

Also Published As

Publication number Publication date
CN115105471B (zh) 2023-01-20
CN115105471A (zh) 2022-09-27

Similar Documents

Publication Publication Date Title
WO2023237124A1 (zh) 一种藏红花素混悬剂及其在制备快速抗抑郁药物中的应用
CN107951869A (zh) 含有大麻二酚的药物制剂及其应用
ES2773834T5 (es) Métodos y composiciones para tratar la depresión usando ciclobenzaprina
US20090264522A1 (en) Methods of treating central nervous system disorders with a low dose combination of escitalopram and bupropion
CN113413461A (zh) 抗老年痴呆药物及其制备方法
CN118593457A (zh) 芬戈莫德在制备治疗缺血性脑卒中口鼻吸入制剂中的应用
CN105919991B (zh) 泽兰素在制备治疗抑郁症药物中的应用
CN114948980B (zh) 一种预防和/或治疗焦虑性失眠的药物组合物及其应用
CN107007590A (zh) 淫羊藿苷元在制备预防或治疗帕金森病药物中的用途
CN105169290A (zh) 一种治疗原发性三叉神经痛的中药组合物及其用途
CN117599076A (zh) 三乙酰基-3-羟基苯基腺苷在制备运动模拟药物中的应用
JP7342259B2 (ja) 過敏性腸症候群の予防又は治療のための医薬的組み合わせ
CN112438994B (zh) 一种对双相情感障碍具有防治作用的小肠粘膜提取物
CN116585332B (zh) 高剂量藏红花素在制备快速抗抑郁药物中的应用
CN101181486A (zh) 一种治疗抑郁症的药物及其制备方法
WO2016041439A1 (zh) 一种柚皮苷与盐酸左西替利嗪药物组合物及其制剂
CN106138072A (zh) 基于积雪草苷的治疗抑郁症复合药物
CN116617313B (zh) 一种兼具抗抑郁、抗焦虑和抗疲劳功效的中药组合物及其制备方法
CN112023027B (zh) 胸腺素或其衍生物的应用和治疗快感缺乏型抑郁症的药物
WO2016041438A1 (zh) 一种柚皮苷与盐酸非索非那丁药物组合物及其制剂
Neerati A textbook of pharmacology-III
KR20240070564A (ko) 알츠하이머병 예방 또는 치료용 약학적 조성물
KR102790590B1 (ko) 지방 분해용 조성물
CN115671106B (zh) 吗啡的应用
CN110448614A (zh) 一种干预特禀体质的中药组合物、破壁组合物及其制备方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23819287

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 23819287

Country of ref document: EP

Kind code of ref document: A1