CN114767690B - Application of cephalothin acid in preparation of antidepressant drug - Google Patents

Application of cephalothin acid in preparation of antidepressant drug Download PDF

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CN114767690B
CN114767690B CN202210712283.7A CN202210712283A CN114767690B CN 114767690 B CN114767690 B CN 114767690B CN 202210712283 A CN202210712283 A CN 202210712283A CN 114767690 B CN114767690 B CN 114767690B
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antidepressant
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CN114767690A (en
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郭秋岩
王继刚
徐承超
王启新
谷丽维
张书杰
唐欢
柴新
杨通
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/54Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
    • A61K31/542Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/545Compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins, cefaclor, or cephalexine
    • A61K31/546Compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins, cefaclor, or cephalexine containing further heterocyclic rings, e.g. cephalothin
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00Drugs for disorders of the nervous system
    • A61P25/24Antidepressants

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Abstract

The invention relates to the technical field of biological medicines, and in particular relates to application of cephalothin acid in preparation of antidepressant medicines. The application of the cephalothin acid in the preparation of antidepressant drugs is provided. The cephalothin acid can effectively relieve PC12 cell injury caused by corticosterone induction and relieve neuroinflammation. The corticosterone-induced depression model can simulate the pathological process of depression, and can be used for the efficacy evaluation and mechanism exploration of antidepressant medicaments. The patent can provide scientific basis for clinical medication of depression.

Description

Application of cephalothin acid in preparation of antidepressant drugs
Technical Field
The invention relates to the technical field of biological medicines, and in particular relates to application of cephalothin acid in preparation of antidepressant medicines.
Background
Depression is a type of mood disorder characterized primarily clinically by a marked and persistent mood drop, the most common affective disorder. It has the characteristics of high disease incidence, high recurrence rate and high disability rate. The world health organization survey in 2021 showed that about 3.5 million people worldwide had depression, which would be the second leading cause of premature death or disability. Depression not only affects the quality of life of patients, but also places a heavy economic burden on patients and families.
Researchers believe that inflammatory cytokines, neurotrophic factors, hypothalamic-pituitary-adrenal Hormone (HPA) axons may be present as biomarkers of depression. At present, one of the accepted theories of the pathogenesis of depression is HPA axial hyperactivity. Corticosterone is an important hormone at the end of the HPA axis, and its abnormal increase can lead to depressive features. Nerve cell damage is one of important indexes of nerve cell apoptosis, and rat adrenal pheochromocytoma cell PC12 is a common nerve cell strain for researching depression and has the general characteristics of neuroendocrine cells. Apoptosis refers to the autonomous, ordered death of cells under the control of genes in order to maintain homeostasis, and a decrease in mitochondrial membrane potential is a marker event in the early stages of apoptosis. Corticosterone-induced poorly differentiated PC12 cells are used as an in vitro research model for depression, apoptosis-related indexes are detected, and the protective effect of the medicine on nerve cells can be observed.
At present, first-line antidepressant drugs mainly comprise fluoxetine, paroxetine, sertraline, fluvoxamine and citalopram, target serotonin and norepinephrine pathways, and can relieve depression symptoms to a certain extent, but have the limitations of easy relapse after stopping treatment and the like. Therefore, it is important to obtain a drug which can alleviate depression by protecting nerve cells with little side effects.
The cephalothin acid is a broad-spectrum sterilization type antibiotic in the breeding period, is stable to penicillinase and has strong gram-positive bacteria resistance. The antibacterial and anti-infective activities of antibiotics have been widely used clinically. In addition, antibiotics have a series of neuroprotective functions, such as prevention of occurrence of nerve infection, prevention of mitochondria-mediated cytochrome C release, prevention of microglial activation, reduction of cytotoxicity due to accumulation of glutamic acid, and the like. Antibiotics have potential therapeutic significance for partial types of neurological diseases and mental disorders, such as neurological damage caused by cerebral ischemia, mental diseases, depression, schizophrenia and the like. Through retrieval, no report related to the antidepressant efficacy of cephalothin acid is found at present.
The Chinese patent application with application publication number CN201010283126.6 discloses a method for synthesizing cephalothin acid by 7-ACA, which takes one or more of n-hexane, cyclohexane and petroleum ether as a crystallization solvent to crystallize to separate out the cephalothin acid, thereby achieving the characteristics of quick drying, light color and high yield, and the treatment effect of the cephalothin acid for resisting depression by inhibiting inflammation is not clear.
Disclosure of Invention
The application aims to provide an application of cephalothin acid in preparation of an antidepressant drug, and aims to solve the problem that a drug capable of relieving depression symptoms by reducing inflammation level and having small side effects is absent in the prior art.
In order to achieve the purpose of the application, the technical scheme adopted by the application is as follows:
in a first aspect, the application provides the use of cephalothin acid in the preparation of an antidepressant medicament.
Further, the cephalothin acid can realize the antidepressant effect by playing the role of protecting nerve cells.
Further, the exerting of the nerve cell protection effect comprises improving the activity of rat adrenal pheochromocytoma PC12 cells.
Further, the exerting the neural cell protection effect comprises reducing the apoptosis rate of the PC12 cell and increasing the mitochondrial membrane potential.
Further, the cephalothin acid is used as the only active ingredient of the antidepressant drug.
Further, the antidepressant drug comprises at least one of a pharmaceutically acceptable salt, a eutectic crystal, a stereoisomer, a prodrug, a solvate and a metabolite of the cephalothin acid.
Further, the antidepressant drug also comprises pharmaceutically acceptable auxiliary materials; wherein the adjuvants comprise at least one of diluent, excipient, filler, binder, humectant, absorption enhancer, surfactant, lubricant, stabilizer, flavoring agent, sweetener, and pigment.
Furthermore, the antidepressant medicament can be prepared into any pharmaceutically acceptable dosage form, and the dosage form comprises any one of emulsion, cream, pills, dripping pills, capsules, granules, powder, liniment, cataplasm, spray, oral liquid, decoction, injection, sustained release preparation or controlled release preparation.
Further, the drug concentration of the cephalothin acid is 0.1-10 nM.
In a second aspect, the present application provides an antidepressant drug comprising cephalothin acid or a pharmaceutically acceptable salt, cocrystal, stereoisomer, prodrug, solvate, metabolite thereof.
The application of cephalothin acid provided by the first aspect of the application in preparing antidepressant drugs. The cephalothin acid can effectively relieve PC12 cell injury caused by corticosterone induction and relieve neuroinflammation. The pathological process of the corticosterone-induced depression model is close to that of clinical patients, the actual curative effect of the antidepressant can be reflected, a new strategy and a clinical research basis are provided for the antidepressant, and a basis is further provided for guiding clinical medication of the depressive disorder.
An antidepressant drug provided in the second aspect of the present application, comprising cephalothin acid or a pharmaceutically acceptable salt, cocrystal, stereoisomer, prodrug, solvate, metabolite thereof. In the using process, the cephalothin acid is applied to the preparation of antidepressant drugs. The cephalothin acid can effectively relieve PC12 cell injury caused by corticosterone induction and relieve neuroinflammation. The pathological process of the corticosterone-induced depression model is close to that of clinical patients, the actual curative effect of the antidepressant can be reflected, a new strategy and a clinical research basis are provided for the antidepressant, and a basis is further provided for guiding clinical medication of the depressive disorder.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
Wherein:
FIG. 1 shows the effect of cephalothin acid on the viability of PC12 cells at different concentrations;
FIG. 2 is a graph of the effect of cephalothin acid on the cell viability of corticosterone-induced PC12 cell injury at different concentrations;
FIG. 3 is a graph showing the effect of cephalothin acid on the rate of corticosterone-induced apoptosis of PC12 cells at various concentrations.
FIG. 4 is a graph showing the effect of cephalothin acid on the rate of PC12 apoptosis induced by corticosterone at different concentrations.
FIG. 5 is a graph showing the effect of cephalothin acid on corticosterone-induced mitochondrial membrane potential of PC12 cells at various concentrations.
FIG. 6 is a graph showing the effect of cephalothin acid on corticosterone-induced mitochondrial membrane potential of PC12 cells at different concentrations.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In a first aspect of the embodiments herein there is provided the use of cephalothin acid in the manufacture of an antidepressant medicament.
The application of cephalothin acid provided by the first aspect of the embodiment of the application in preparing antidepressant drugs. The cephalothin acid can effectively relieve PC12 cell injury caused by corticosterone induction and relieve neuroinflammation. The pathological process of the corticosterone-induced depression model is close to that of clinical patients, the actual curative effect of the antidepressant can be reflected, a new strategy and a clinical research basis are provided for the antidepressant, and a basis is further provided for guiding clinical medication of the depressive disorder.
In some embodiments, cephalothin acid has the formula C 16 H 16 N 2 O 6 S 2 The molecular weight is 396.438, and the structure is shown in formula (I):
Figure 378506DEST_PATH_IMAGE001
(I)。
in some embodiments, the cephalothin acid achieves antidepressant effect by exerting a neuronal protective effect.
In some embodiments, the exerting a neuroprotective effect comprises increasing viability of rat adrenal pheochromocytoma PC12 cells.
In some embodiments, the exerting a neuroprotective effect comprises decreasing the rate of apoptosis of PC12 cells and increasing mitochondrial membrane potential.
In some embodiments, the cephalothin acid is the only active ingredient of the antidepressant drug.
In some embodiments, the antidepressant drug comprises at least one of a pharmaceutically acceptable salt, co-crystal, stereoisomer, prodrug, solvate, metabolite of cephalothin acid.
In some embodiments, the antidepressant further comprises a pharmaceutically acceptable excipient; wherein the adjuvants comprise at least one of diluent, excipient, filler, binder, humectant, absorption enhancer, surfactant, lubricant, stabilizer, flavoring agent, sweetener, and pigment.
In some embodiments, the antidepressant drug may be formulated into any one of pharmaceutically acceptable dosage forms, including any one of emulsion, cream, pill, drop pill, capsule, granule, powder, liniment, cataplasm, spray, oral liquid, decoction, injection, sustained-release preparation or controlled-release preparation.
In some embodiments, the drug concentration of the cephalothin acid is 0.1-10 nM.
In a second aspect of the embodiments herein there is provided an antidepressant drug comprising cephalothin acid or a pharmaceutically acceptable salt, cocrystal, stereoisomer, prodrug, solvate, metabolite thereof.
An antidepressant drug as provided in the second aspect of the embodiments of the present application, comprising cephalothin acid or a pharmaceutically acceptable salt, cocrystal, stereoisomer, prodrug, solvate, metabolite thereof. In the using process, the cephalothin acid is applied to the preparation of antidepressant drugs. The cephalothin acid can effectively relieve PC12 cell injury caused by corticosterone induction and relieve neuroinflammation. The pathological process of the corticosterone-induced depression model is close to that of clinical patients, can reflect the real curative effect of depression resistance, provides a new strategy and clinical research basis for depression resistance, and further provides a basis for guiding clinical medication of depression.
Specific examples are provided below for illustration.
Cell lines used in the examples: PC12 cells were purchased from Biotech limited, Jones and Xinzhou, Shanghai, by culturing in RPMI-1640 medium containing 10% fetal bovine serum and 1% double antibody at 37 deg.C and 5% CO 2 Culturing in a cell culture box.
The drugs used in the examples: cephalothin acid, provided by Shanghai-sourced leaf Biotechnology Limited, product batch number: B12O10D 99935.
Example 1
(one)) experimental method
The used corticosterone-induced PC12 cell injury model is a well-known ideal external model of depression, simulates the pathological state of depression, and is suitable for inspecting the antidepressant action characteristics of cephalothin acid.
The invention adopts a CCK8 method to evaluate the protective activity of cephalothin acid on PC12 cells damaged by corticosterone. PC12 cells are digested with pancreatin and then cultured in complete medium to obtain cells with a cell content of 1 × 10/ml 5 The suspension of (4) was inoculated into a 96-well plate at 100. mu.L/well, and placed at 37 ℃ in 5% CO 2 And (5) incubator culture. Removing cell sap after 24h, adding a blank group into a complete culture medium, adding a dosing group into a complete culture medium with the concentration of 0.1, 0.25, 0.5, 1, 2.5, 5 and 10nM cephalothin acid respectively, and setting 6 multiple holes for each concentration; after 24h incubation, 10. mu.L of CCK8 solution was added to each well, and after further 1.5 h incubation, the absorbance (OD) of each well was measured at 450nm using a microplate reader, and the cell viability of each group was calculated.
After 24h of plating culture, removing cell sap, adding complete culture medium into a blank group and a model group, adding complete culture medium with the concentrations of 0.25, 0.5, 1, 2.5, 5 and 10nM cephalothin acid into an administration group, and setting 6 multiple holes for each concentration; after 24h incubation, the medium was discarded, the blank group was added with complete medium, and the model group and the administration group were added with complete medium containing 150 μ M corticosterone; after 24h incubation, 10. mu.L of CCK8 solution was added to each well, and after further 1.5 h incubation, the absorbance (OD) of each well was measured at 450nm using a microplate reader, and the cell viability of each group was calculated.
PC12 cells are digested with trypsin and cultured in a complete medium to obtain a cell content of 1 × 10/ml 5 The suspension of (4) was inoculated into a 24-well plate at 500. mu.L/well, and placed at 37 ℃ in 5% CO 2 And (5) incubator culture. After 24h, cell sap is discarded, complete culture media are added into the blank group and the model group, complete culture media with the concentrations of 2.5, 5 and 10nM cephalothin acid are added into the administration group, and 3 multiple wells are arranged at each concentration. And after 24h of incubation, removing the culture medium, adding the complete culture medium into a blank group, adding the complete culture medium containing 150 mu M corticosterone into a model group and an administration group, after 24h of incubation, digesting the cells with pancreatin, collecting the cells, centrifuging for 5 min at 1000 g, removing the supernatant, adding PBS (phosphate buffer solution) for washing once, centrifuging for 5 min at 1000 g, removing the supernatant, adding a binding solution into the cell precipitate for resuspension, then adding an Annexin V-FITC/PI double staining agent in the dark, incubating for 15 min in the dark at room temperature, and detecting the apoptosis rate on a computer.
After cell precipitation is prepared as above, JC-1 staining working solution is added, incubation is carried out for 20 min at 37 ℃, centrifugation is carried out for 4 min at 600 g, cells are collected, 1 mL of JC-1 staining buffer solution is added for washing twice, the cells are precipitated, 200 μ L of JC-1 staining buffer solution is added for resuspension, and mitochondrial membrane potential is detected on a machine.
(II) results of the experiment
2.1 the cephalothin acid concentration is not higher than 10nM and has no obvious toxicity to PC12 cell.
The effect of cephalothin acid on the activity of PC12 cells is shown in FIG. 1. The result shows that when the concentration is not higher than 10nM, the cephalothin acid has no obvious cytotoxicity to PC12 cells, and provides a safe range reference for the use concentration of the cephalothin acid in subsequent experiments.
2.2 the cephalothin acid can obviously improve PC12 cell damage induced by corticosterone.
The protective effect of cephalothin acid on PC12 cells injured by corticosterone is shown in FIG. 2 (C: (C)) ### P<0.001vs blank; *** P<0.001vs model set). The results show that the cell survival rate of the administration group at each concentration is improved to a certain extent compared with that of the model group, and statistical differences exist, which indicates that the compound can effectively relieve corticosterone-induced PC12 cell injury.
2.3 the cephalothin acid can obviously reduce the PC12 cell apoptosis rate induced by corticosterone.
The effect of cephalothin acid on the rate of corticosterone-induced apoptosis of PC12 cells is shown in FIGS. 3 and 4: ( ### P<0.001vs blank; * P<0.05 vs model set). Compared with a blank control group, the apoptosis rate of the model group is obviously increased; compared with the model group, the apoptosis rate of the cephalothin acid administration group is obviously reduced, and the 5nM concentration shows obvious difference; the application of the cephalothin acid drug treatment has obvious anti-apoptosis effect on PC12 cells damaged by corticosterone.
2.4 Cefthiophene acid can significantly reverse corticosterone-induced decrease in mitochondrial membrane potential of PC12 cells.
The effect of cephalothin acid on corticosterone-induced mitochondrial membrane potential of PC12 cells is shown in fig. 5 and 6. ( ### P<0.001vs blank; ** P<0.01 vs model set). Compared with a blank control group, the model group has the advantages that the cell mitochondrial membrane potential is obviously reduced; compared with a model group, the mitochondrial membrane potential of the cephalothin acid administration group is obviously increased, and the concentration of 10nM shows obvious difference; the method shows that the early apoptosis of PC12 cells damaged by corticosterone can be improved by adopting cephalothin acid for treatment.
In conclusion, the application of the cephalothin acid provided by the application in preparing the antidepressant drug is provided. The cephalothin acid can effectively relieve PC12 cell injury caused by corticosterone induction and relieve neuroinflammation. The corticosterone-induced depression model can simulate the pathological process of depression, and can be used for the efficacy evaluation and mechanism exploration of antidepressant medicaments. The patent can provide scientific basis for clinical medication of depression.
The above disclosure is only for the purpose of illustrating the preferred embodiments of the present invention, and it is therefore to be understood that the invention is not limited by the scope of the appended claims.

Claims (6)

1. The application of cephalothin acid in preparing antidepressant drugs is disclosed, wherein the structure of cephalothin acid is shown as formula I:
Figure 845989DEST_PATH_IMAGE001
formula I; the cephalothin acid is used as the only active ingredient of the antidepressant drug; the cephalothin acid realizes the anti-depression effect by playing the role of protecting nerve cells; wherein the drug concentration of the cephalothin acid is 0.1-10 nM.
2. The use of claim 1, wherein said neuroprotective effect comprises increasing the viability of rat adrenal pheochromocytoma PC12 cells.
3. The use of claim 1, wherein the neuroprotective effect comprises decreasing the rate of apoptosis of PC12 cells and increasing mitochondrial membrane potential.
4. The use according to any one of claims 1 to 3, wherein the antidepressant comprises cephalothin acid and pharmaceutically acceptable salts thereof.
5. The use of any one of claims 1 to 3, wherein the antidepressant further comprises a pharmaceutically acceptable excipient; wherein the adjuvants comprise at least one of diluent, excipient, filler, binder, humectant, absorption enhancer, surfactant, lubricant, stabilizer, flavoring agent, sweetener, and pigment.
6. The use of any one of claims 1 to 3, wherein the antidepressant is formulated into any one of pharmaceutically acceptable dosage forms, including emulsion, cream, pill, drop pill, capsule, granule, powder, liniment, cataplasm, spray, oral liquid, decoction, injection, sustained release preparation or controlled release preparation.
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5977090A (en) * 1996-09-27 1999-11-02 Guilford Pharmaceuticals Inc. Pharmaceutical compositions and methods of treating compulsive disorders using NAALADase inhibitors
CN1871243A (en) * 2003-08-25 2006-11-29 雷瓦尔克斯药品有限公司 Oral neurotherapeutic cefazolin compositions
CN101979393A (en) * 2010-09-16 2011-02-23 苏州中联化学制药有限公司 Method for synthesizing cephalothin acid by using 7-aminocephalosporanic acid (ACA)
CN104151242A (en) * 2014-06-06 2014-11-19 四川大学 Dihydro isoquinoline compound and application thereof in preparation of nerve protection or antidepressant medicament
CN105820053A (en) * 2016-04-23 2016-08-03 黄亦琼 Pharmaceutical composition of cefalexin and application of pharmaceutical composition in biological medicine

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Publication number Priority date Publication date Assignee Title
EP1212055B1 (en) * 1999-08-16 2011-08-03 Revaax Pharmaceuticals LLC Neurotherapeutic composition comprising a beta-lactam compound
US8101600B2 (en) * 2008-12-23 2012-01-24 Brookhaven Science Associates, Llc Method of treating depression

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5977090A (en) * 1996-09-27 1999-11-02 Guilford Pharmaceuticals Inc. Pharmaceutical compositions and methods of treating compulsive disorders using NAALADase inhibitors
CN1871243A (en) * 2003-08-25 2006-11-29 雷瓦尔克斯药品有限公司 Oral neurotherapeutic cefazolin compositions
CN101979393A (en) * 2010-09-16 2011-02-23 苏州中联化学制药有限公司 Method for synthesizing cephalothin acid by using 7-aminocephalosporanic acid (ACA)
CN104151242A (en) * 2014-06-06 2014-11-19 四川大学 Dihydro isoquinoline compound and application thereof in preparation of nerve protection or antidepressant medicament
CN105820053A (en) * 2016-04-23 2016-08-03 黄亦琼 Pharmaceutical composition of cefalexin and application of pharmaceutical composition in biological medicine

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