CN113292524B - Butylphthalide derivative and application thereof in preparation of medicine for protecting nerve cells - Google Patents

Butylphthalide derivative and application thereof in preparation of medicine for protecting nerve cells Download PDF

Info

Publication number
CN113292524B
CN113292524B CN202110846974.1A CN202110846974A CN113292524B CN 113292524 B CN113292524 B CN 113292524B CN 202110846974 A CN202110846974 A CN 202110846974A CN 113292524 B CN113292524 B CN 113292524B
Authority
CN
China
Prior art keywords
butylphthalide
acid
cells
derivative
disease
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.)
Active
Application number
CN202110846974.1A
Other languages
Chinese (zh)
Other versions
CN113292524A (en
Inventor
秦川
李彦红
梁唯
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.)
Institute of Laboratory Animal Science of CAMS
Original Assignee
Institute of Laboratory Animal Science of CAMS
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 Institute of Laboratory Animal Science of CAMS filed Critical Institute of Laboratory Animal Science of CAMS
Priority to CN202110846974.1A priority Critical patent/CN113292524B/en
Publication of CN113292524A publication Critical patent/CN113292524A/en
Application granted granted Critical
Publication of CN113292524B publication Critical patent/CN113292524B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
    • C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D307/87—Benzo [c] furans; Hydrogenated benzo [c] furans
    • C07D307/88—Benzo [c] furans; Hydrogenated benzo [c] furans with one oxygen atom directly attached in position 1 or 3
    • 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
    • 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
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • 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
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs
    • 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
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia

Abstract

The invention relates to the fields of biological medicine and chemistry, and provides a butylphthalide derivative and application thereof in preparing a medicament for protecting nerve cells, in particular to application of the butylphthalide derivative or a medicinal composition consisting of the butylphthalide derivative in preparing a medicament for preventing and/or treating diseases related to cell injury and neurodegenerative diseases; the nerve cell damage diseases include but are not limited to stroke, spinal cord injury and nervous system diseases accompanied with blood brain barrier injury; the neurodegenerative disease includes but is not limited to Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, spinal muscular atrophy, primary lateral sclerosis, or spinocerebellar ataxia.

Description

Butylphthalide derivative and application thereof in preparation of medicine for protecting nerve cells
Technical Field
The invention relates to the fields of biological medicine and chemistry, in particular to a butylphthalide derivative and application thereof in preparing a medicament for protecting nerve cells.
Background
Butylphthalide is a small molecular protein extracted from celery or celery seeds. This protein is small and can easily pass through the blood-brain barrier to exert its effect on brain cells. In 1978, the institute of medicine of Chinese academy of medicine sciences separated levo-butylphthalide from cress seeds. Butylphthalide is first chemically synthesized in 1980, the initial research direction of the butylphthalide is anti-epilepsy, but the butylphthalide has larger potential safety hazard due to the fact that the therapeutic dose for resisting epilepsy is close to the toxic dose, and the research is accordingly stranded.
The butylphthalide is mainly used for treating mild and moderate acute ischemic stroke, can reduce the concentration of calcium ions in cells, inhibit the release of glutamic acid, reduce the generation of arachidonic acid, clear oxygen free radicals, improve the activity of oxidase and the like, and acts on a plurality of pathological links of cerebral ischemia. Meanwhile, the composition has a strong anti-cerebral ischemia effect, can obviously improve microcirculation and blood flow of cerebral ischemia areas, increase the number of capillaries in the cerebral ischemia areas, relieve cerebral edema, reduce the volume of cerebral infarction, improve brain energy metabolism and also has a certain effect of inhibiting thrombus. The earlier in onset, the better the prognosis for the patient. The butylbenzene peptide can take effect within 24 hours physiologically, but if the patient needs to have obvious improvement of symptoms clinically, at least 5-7 days are needed.
All the medicines have positive and side effects, and the butylphthalide has the same side effect as the positive effect, and can activate brain cells and promote the metabolism of the brain cells. When the brain cells are improperly activated after activation, hallucinations and epilepsy may occur. In addition, all drugs are metabolized by liver and kidney functions, and there may be some patients with elevated transaminase. Since it increases blood flow in brain and increases proliferation of blood vessels, it can produce blood seepage at cerebral infarction area in case of some special cerebral infarction or few cerebral infarction.
Disclosure of Invention
In one aspect, the invention provides butylphthalide derivatives and pharmaceutically acceptable salts thereof, as shown in the following general formulae.
Figure DEST_PATH_IMAGE001
Wherein R is1、R2Each independently may be an atom or a substituent.
In one embodiment, the atoms include hydrogen atoms or halogen atoms; preferably, the halogen atom includes a fluorine atom, a chlorine atom, a bromine atom, an iodine atom.
In one embodiment, the substituents include, but are not limited to, the following substituents and heteroatom-containing substituents that are common: alkyl, carboxyl, sulfonic acid group, hydrocarbonoxy carbonyl, formyl, haloformyl, oxo, carbamoyl, cyano, phenolic alkyl, phenolic hydroxyl, alcoholic hydroxyl, amino, hydrocarbonoxy, alkyl, nitro, nitroso; the substituents are alsoMay include C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Cycloalkyl, aryl, heteroaryl, heterocyclyl- (CH)2) n-, aryl-C1-6Alkyl-, heteroaryl-C1-6Alkyl-, aryl- (CH)2) n-O-, heteroaryl- (CH)2)n-O-、C3-8cycloalkyl-C (O) -, heterocyclyl-C (O) -, aryl-C (O) -, or heteroaryl-C (O) -, wherein C1-6Alkyl radical, C1-6Alkoxy radical, C3-8Cycloalkyl, aryl, heterocyclyl- (CH)2) n-, aryl-C1-6Alkyl-, heteroaryl-C1-6Alkyl-, aryl- (CH)2) n-O-, heteroaryl- (CH)2)n-O-、C3-8cycloalkyl-C (O) -, heterocyclyl-C (O) -, aryl-C (O) -, or heteroaryl-C (O, hydroxy, mercapto, amino, nitro, acyl, silyl, acyloxy, oxyacyl, borono, hydroxyamino, nitroso, silyl.
In one embodiment, the heteroatoms include nitrogen, oxygen, sulfur, phosphorus, boron, chlorine, bromine, iodine.
In one embodiment, the number of heteroatoms is from 1 to 6; specifically, 1, 2, 3, 4, 5, 6 are included; more preferably, 1-3.
In one embodiment, the substituent may or may not contain carbon atoms.
In one embodiment, the substituent contains a carbon atom, the number of which is not particularly limited; preferably, 1 to 20 carbon atoms; more preferably, 1 to 10 carbon atoms.
In one embodiment, the carbon atom-containing substituent comprises hydrocarbyl, substituted hydrocarbyl, phenyl, butylphenyl, p-methylphenyl, trifluoromethyl, 2, 2, 2-trifluoroethyl, methyl, ethyl, n-propyl, isopropyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, benzyl, substituted C1-20Alkyl, substituted aryl, substituted C1-20An open chain heterocarbon group,Substituted heteroaryl hydrocarbon groups, and the like.
In one embodiment, the substituted hydrocarbyl group is a 1 carbon atom-containing group substituted with 1 to 3 heteroatoms.
In one embodiment, the substituted hydrocarbyl is trifluoromethyl, difluoromethyl, monofluoromethyl, tribromomethyl, dibromomethyl, bromomethyl, trichloromethyl, dichloromethyl, chloromethyl, iodomethyl, diiodomethyl.
In one embodiment, the substituted hydrocarbyl group is trifluoromethyl.
In one embodiment, the substituent free of carbon atoms includes hydroxyl, mercapto, amino, nitro, acyl, silyl, acyloxy, oxyacyl, boroxy, hydroxyamino, nitroso, silyl.
In one embodiment, the substituent containing no carbon atom is a nitroso group, a nitro group.
In one embodiment, the substituent containing no carbon atoms is a nitro group.
In one embodiment, the R is1、R2Selected from any one atom or substituent of hydrogen atom, nitro or trifluoromethyl.
In one embodiment, the R is1Selected from hydrogen atom, nitro or trifluoromethyl, the R2Selected from a hydrogen atom or a trifluoromethyl group.
In one embodiment, the derivatives of butylphthalide include: 3-butyl-7-nitroisobenzofuran, 3-butyl-7- (trifluoromethyl) isobenzofuran, 3-butyl-5- (trifluoromethyl) isobenzofuranone.
In one embodiment, the 3-butyl-7-nitroisobenzofuran has the formula:
Figure 669323DEST_PATH_IMAGE002
in one embodiment, the 3-butyl-7- (trifluoromethyl) isobenzofuran has the formula:
Figure DEST_PATH_IMAGE003
in one embodiment, the 3-butyl-5- (trifluoromethyl) isobenzofuranone has the formula:
Figure 914360DEST_PATH_IMAGE004
in the present invention, the pharmaceutically acceptable salts include salts derived from inorganic acids and organic acids.
Preferably, the inorganic acid may be hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid.
Preferably, the organic acid may be an amino acid, methanesulfonic acid, trifluoromethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, 1-naphthalenesulfonic acid, 2-naphthalenesulfonic acid, acetic acid, trifluoroacetic acid, malic acid, tartaric acid, citric acid, oxalic acid, succinic acid, fumaric acid, maleic acid, benzoic acid, salicylic acid, phenylacetic acid or mandelic acid, camphoric acid, glutaric acid, oxalic acid, lactic acid or malonic acid, pamoic acid, hydroxynaphthoic acid, gentisic acid, glycolic acid, mandelic acid, 4-acetamidobenzoic acid or nicotinic acid; wherein the amino acid comprises glycine, alanine, lysine, arginine, serine, phenylalanine, proline, tyrosine, aspartic acid, glutamic acid, histidine, leucine, methionine, threonine, pyroglutamic acid, tryptophan or valine.
In another aspect, the present invention provides a pharmaceutical composition comprising the butylphthalide derivative of the present invention as an active ingredient. The pharmaceutical composition may be prepared according to methods well known in the art.
In one embodiment, the pharmaceutical composition may be a tablet (including sugar-coated tablets, film-coated tablets, sublingual tablets, orally disintegrating tablets, buccal tablets, and the like), pill, powder, granule, capsule (including soft capsules, microcapsules), lozenge, syrup, liquid, emulsion, suspension, controlled release formulation (e.g., immediate release formulation, sustained release microcapsule), aerosol, film (e.g., orally disintegrating film, oral mucosa-adhesive film), injection (e.g., subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection), intravenous drip, transdermal absorption formulation, ointment, lotion, adhesive formulation, suppository (e.g., rectal suppository, vaginal suppository), pellet, nasal formulation, pulmonary formulation (inhalation), eye drop, and the like, oral or parenteral formulation (e.g., intravenous, intramuscular, subcutaneous, intraorgan, intranasal, intradermal, instillation, intracerebral, intrarectal, etc., administration to the vicinity of tumors and directly to lesions).
In one embodiment, a pharmaceutically acceptable solid/liquid excipient is also included in the pharmaceutical composition.
In one embodiment, the pharmaceutically acceptable solid/liquid excipients include binders, fillers, lubricants, and disintegrants.
In one embodiment, the binder includes, but is not limited to, corn starch, potato starch or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethylcellulose calcium, carboxymethylcellulose sodium), polyvinylpyrrolidone, methyl cellulose, pregelatinized starch, hydroxypropylmethylcellulose microcrystalline cellulose, and mixtures thereof.
In one embodiment, the filler includes, but is not limited to, talc, calcium carbonate (e.g., granules or powder), lactose, microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof.
In one embodiment, the lubricant includes, but is not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerol, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, and mixtures thereof.
In one embodiment, the disintegrant includes, but is not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pregelatinized starch, other starches, clays, other algins, other celluloses, gums, and mixtures thereof.
In one embodiment, the pharmaceutical composition of the present invention may be administered by any of the following means: oral, aerosol inhalation, rectal, nasal, buccal, topical, parenteral, such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intracardiac, intrasternal or intravenous administration.
In one embodiment, the pharmaceutical composition of the present invention can be administered alone or in combination with other agents for treating neurological-related disorders. Preferably, the nerve-related disease medicament comprises traditional Chinese medicines, western medicines or bioactive preparations.
In one embodiment, the animal to be treated comprises a mammal, reptile, crustacean, amphibian, fish, poultry. The main range is mammalian, especially human.
In another aspect, the invention provides a use of the derivative or the pharmaceutical composition of butylphthalide in preparation of a medicament for preventing and/or treating cell injury related diseases and neurodegenerative diseases.
In one embodiment, the cell comprises an adipocyte, inflammatory cell, endothelial cell, epithelial cell, neural cell, stem cell, lymphocyte.
In one embodiment, the neural cell comprises a neuron, a glial cell, a neuroblast; more specifically, human neuroblastoma cells; more specifically, the SHY5Y cell line.
In one embodiment, the neuronal cell injury disorder includes, but is not limited to, stroke, spinal injury, and neurological disorders associated with blood brain barrier damage.
In one embodiment, the neurodegenerative disease includes, but is not limited to, parkinson's disease, alzheimer's disease, huntington's disease, amyotrophic lateral sclerosis, spinal muscular dystrophy, primary lateral sclerosis, or spinocerebellar ataxia.
Drawings
FIG. 1 is a graph showing the results of quality inspection (HNMR) s of 3-butyl-7-nitroisobenzofuran.
FIG. 2 is a graph showing the results of quality control (HNMR) s of 3-butyl-7- (trifluoromethyl) isobenzofuran.
FIG. 3 is a graph showing the results of quality inspection (HNMR) s of 3-butyl-5-bromoisobenzofuranone.
FIG. 4 is a graph showing the results of (HNMR) s quality inspection of 3-butyl-5- (trifluoromethyl) isobenzofuranone.
FIG. 5 shows the Butylphthalide derivative vs. Abeta25-35Graph of the results of MTT activity assay performed on treated SY5Y cells.
FIG. 6 shows the Butylphthalide derivative vs. Abeta25-35Graph of the results of injury reactive oxygen species release assay performed on treated SY5Y cells.
FIG. 7 is a graph showing the results of LDH release assay of butylphthalide derivatives on SY5Y cells cultured in the absence of sugar and oxygen.
Detailed Description
The present invention will be further described with reference to the following examples, which are intended to be illustrative only and not to be limiting of the invention in any way, and any person skilled in the art can modify the present invention by applying the teachings disclosed above and applying them to equivalent embodiments with equivalent modifications. Any simple modification or equivalent changes made to the following embodiments according to the technical essence of the present invention, without departing from the technical spirit of the present invention, fall within the scope of the present invention.
Example 1 Synthesis of derivatives
(ii) 3-butyl-7-nitroisobenzofuran
The chemical formula is as follows:
Figure DEST_PATH_IMAGE005
the synthetic route is as follows:
Figure 959676DEST_PATH_IMAGE006
FIG. 1 shows the quality inspection results.
3-butyl-7- (trifluoromethyl) isobenzofuran
The chemical formula is as follows:
Figure DEST_PATH_IMAGE007
the synthetic route is as follows:
Figure 417202DEST_PATH_IMAGE008
FIG. 2 shows the quality inspection results.
3-butyl-5-bromoisobenzofuranone
The chemical formula is as follows:
Figure 762733DEST_PATH_IMAGE009
the synthetic route is as follows:
Figure DEST_PATH_IMAGE010
FIG. 3 shows the quality inspection results.
3-butyl-5- (trifluoromethyl) isobenzofuranone
The chemical formula is as follows:
Figure 146047DEST_PATH_IMAGE011
the synthetic route is as follows:
Figure 209818DEST_PATH_IMAGE012
FIG. 4 shows a quality control result chart.
The synthesis of the compounds is qualified, and can be applied to the next experiment.
Example 2 assay of Activity of Butylphthalide derivatives on anti-apoptotic MTT of A β -induced SY5Y neural cells
The experimental steps are as follows:
SHY5Y cells at 1 x105Per cm2The culture was carried out in 96-well plates at a density of 24 h.
2. When the confluence degree of the cells reaches 80% -90%, the culture medium is discarded, the cells are washed for 3 times by PBS, the drugs (0.5 mM, diluted by normal medium DMEM) are respectively added, and the cells are cultured for 4 hours at 37 ℃.
3. The drug was discarded, washed 2 times with PBS, and 20. mu.M of Abeta was added thereto25-35(PBS is added in advance for dissolution, and the mixture is incubated for 2 days at 37 ℃) and the mixture is incubated for 48 hours at 37 ℃ with the drug, wherein A beta25-35Groups served as model controls. PBS groups (marginal wells filled with sterile PBS to eliminate marginal effects) and blank controls (normal cells cultured in normal medium without any treatment) were included.
4. MTT solution (5 mg/mL) in 10. mu.L PBS was added to each well and incubated at 37 ℃ for 4 h. The medium was carefully aspirated out using a 1mL syringe, 150. mu.L of LDMSO was added to each well, and after fully shaking to completely dissolve the formazan crystals, the absorbance was measured at 570nm using a microplate reader. Meanwhile, a PBS group and a blank cell group cultured by a serum-free culture medium are respectively set as positive control and negative control.
5. Percent active cells in each group = (each group value-PBS group mean)/(blank group mean-PBS mean) × 100%
The experimental results are as follows:
the results of MTT assay are shown in FIG. 5: a novel compound, namely a butylphthalide derivative (3-butyl-7-nitroisobenzofuran); the 3-butyl-7- (trifluoromethyl) isobenzofuran has relatively high activity value, is superior to Butylphthalide (BNP) and levogyration (BNP-L, BNP-R) compounds, is superior to 3-butyl-5 bromo isobenzofuranone, and has statistical significance in difference. The activity values of the two compounds are not obviously different from those of 3-butyl-5- (trifluoromethyl) isobenzofuranone, and the activity value of the two compounds is higher than that of a model group and a BNP group, but is not different from that of BNP-L and BNP-R. The compounds I, II and III have better anti-nerve cell apoptosis activity, but the compounds I and II are relatively better;
on all pictures of the invention: abeta is cell treated by Abeta 25-35, BNP-R refers to D-BNP, BNP-L refers to L-BNP, BNP is racemic BNP, 7F-BNP refers to butylphthalide derivative (7 NO)2BNP refers to butylphthalide derivative (r), 5Br-BNP refers to butylphthalide derivative (r), and 5F-BNP refers to butylphthalide derivative (r).
Example 3 Butylphthalide derivative active oxygen Release test for A β -induced SY5Y nerve cell injury
According to the MTT experiment result, the nerve cell injury active oxygen release experiment is further detected, and all compounds, but butylphthalide and left-right-handed and third-handed cells are dead and do not participate in analysis.
The experimental steps are as follows:
SHY5Y cells at 1 x105Per cm2The culture was carried out in 96-well plates at a density of 24 h.
2. When the confluence degree of the cells reaches 80% -90%, the culture medium is discarded, the cells are washed for 3 times by PBS, and medicines (0.5 mM, normal culture medium DMEM is diluted) are respectively added to the cells to be cultured for 4 hours at 37 ℃.
3. The drug was discarded, washed 2 times with PBS, and 20. mu.M of Abeta was added thereto25-35(PBS was added in advance for dissolution, and incubated at 37 ℃ for 2 days) and the drug mixture was incubated at 37 ℃ for 48h, in which A.beta.was added25-35Groups served as model controls. PBS groups (marginal wells filled with sterile PBS to eliminate marginal effects) and blank controls (normal cells cultured in normal medium without any treatment) were included.
4. And after the incubation is finished, performing ROS detection according to an ROS kit.
PBS washing for 2 times, adding probe, incubating at 37 deg.C for 20min (diluting DCFH-DA with serum-free culture solution according to the instruction 1:1000 to make the final concentration 10. mu.M), and mixing by reversing every 3-5 min to make the probe and the cells fully contact.
And 6, washing the cells for 3 times by PBS, and detecting the light absorption value by a fluorescence microplate reader (488 nm excitation wavelength and 525nm emission wavelength).
ROS activity value = (group mean-PBS group mean)/(blank group mean-PBS group mean) experimental results:
the results are shown in FIG. 6: the ROS activity value is lowest, and the ROS activity value is statistically different from other groups. The activity values of the compounds (II) and (IV) are lower than that of a model group, and both have statistical difference. ② and (iv) have no statistical difference. The activity of the compounds (i), (ii) and (iv) against neuronal damage was further verified, preferably (i). The MTT experiment is integrated, and the first and the second are better.
Example 4 Butylphthalide derivatives glucose deprivation and hypoxia for SY5Y nerve cells for 12 hours, LDH Release test
The experimental steps are as follows:
SHY5Y cells at 1 x105Per cm2The culture was carried out in 24-well plates at a density of 24 h.
2. When the confluence degree of the cells reaches 80-90%, the culture medium is discarded, the cells are washed for 3 times by PBS, medicines of different compounds (diluted by normal DMEM) are respectively added, and the cells are cultured for 4 hours at 37 ℃.
3. The drugs were discarded, washed 2 times with PBS, added with different concentrations of drugs (low-sugar DMEM dilution), and the periphery of the plate was sealed with tape and incubated at 37 ℃ for 12 h. The blank group is in normal culture medium and is not closed; the model groups were closed with low sugar medium and negative and positive controls were performed, respectively.
4. After the incubation, the cells were washed 3 times with PBS, digested with 10. mu.L of trypsin per well, resuspended and centrifuged with 200. mu.L of LPBS, and then detected using LDH kit.
5. Press 5x1061ml of the extract was added to each cell. Ultrasonically crushing cells (ice bath, power 20% or 200w, ultrasonic for 3s, interval of 10s, repeated for 30 times); centrifuging at 8000g and 4 deg.C for 10min, collecting supernatant, and testing on ice.
6. Adding the sample according to the instruction, mixing, performing water bath at 25 deg.C for 15min, adding the sample, mixing, and performing water bath at 255 deg.C for 15 min; adding sample, mixing, standing for 3min, placing 200 μ l in 96-well plate, and measuring absorbance at 450 nm.
7. And (5) making a standard curve according to the concentration and the absorbance value of the standard substance to obtain a formula. And calculating the LDH activity value according to a formula.
The experimental results are as follows:
the results are shown in FIG. 7: the novel medicament has the advantages that the novel medicament has LDH release experiments on the glucose-deficient hypoxia injury protection activity of nerve cells, and found that butylphthalide and levorotatory cells are dead more and do not participate in comparative analysis.
The LDH activity values of the compounds I and II are small relative to the model group, and both have statistical difference. The compound (iv) is lower than the model group but has no obvious difference, and the compound (iii) is higher than the model group and has statistical difference, and the effect is the worst.
The butylphthalide derivatives I and II have excellent protective activity on the 12-hour injury of the nerve cells due to sugar deficiency and oxygen deficiency.

Claims (4)

1. The derivatives of butylphthalide or pharmaceutically acceptable salts thereof, wherein the derivatives of butylphthalide are any one or more of 3-butyl-7-nitroisobenzofuran, 3-butyl-7- (trifluoromethyl) isobenzofuran and 3-butyl-5- (trifluoromethyl) isobenzofuranone.
2. A pharmaceutical composition comprising the butylphthalide derivative according to claim 1, or a pharmaceutically acceptable salt thereof.
3. Use of the butylphthalide derivative or pharmaceutically acceptable salt thereof according to claim 1 and the pharmaceutical composition according to claim 2 for preparing a medicament for preventing and/or treating cell injury related diseases and neurodegenerative diseases.
4. The use of claim 3, wherein the disease associated with cell damage comprises a disease associated with nerve cell damage.
CN202110846974.1A 2021-07-27 2021-07-27 Butylphthalide derivative and application thereof in preparation of medicine for protecting nerve cells Active CN113292524B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110846974.1A CN113292524B (en) 2021-07-27 2021-07-27 Butylphthalide derivative and application thereof in preparation of medicine for protecting nerve cells

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110846974.1A CN113292524B (en) 2021-07-27 2021-07-27 Butylphthalide derivative and application thereof in preparation of medicine for protecting nerve cells

Publications (2)

Publication Number Publication Date
CN113292524A CN113292524A (en) 2021-08-24
CN113292524B true CN113292524B (en) 2021-10-08

Family

ID=77331003

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110846974.1A Active CN113292524B (en) 2021-07-27 2021-07-27 Butylphthalide derivative and application thereof in preparation of medicine for protecting nerve cells

Country Status (1)

Country Link
CN (1) CN113292524B (en)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114478450A (en) * 2022-01-10 2022-05-13 四川大学 Benzyloxybelphthalide compound, preparation method and application thereof
CN114773330B (en) * 2022-05-24 2024-03-15 贵州医科大学 Butylphthalide-oxa/thiadiazole compound and preparation method and application thereof
WO2024061367A1 (en) * 2022-09-23 2024-03-28 山东如至生物医药科技有限公司 Novel compound, pharmaceutical composition thereof, and use thereof

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100554259C (en) * 2007-04-12 2009-10-28 郑州大学 2-benzo [c] furan ketone compound and application thereof
CN112794831B (en) * 2021-04-06 2021-07-27 北京理工大学 3- (3' -hydroxybutyl) isobenzofuran-1 (3H) -one derivative, composition, preparation method and application thereof

Also Published As

Publication number Publication date
CN113292524A (en) 2021-08-24

Similar Documents

Publication Publication Date Title
CN113292524B (en) Butylphthalide derivative and application thereof in preparation of medicine for protecting nerve cells
JP7470151B2 (en) Use of chloroquine and clemizole compounds to treat inflammatory and cancerous conditions
US7888355B2 (en) Compositions and methods for inhibiting growth and metastasis of melanoma
US20040248983A1 (en) N-arylphenylacetamide derivatives and medicinal compositions containing the same
KR20100100584A (en) Isosorbide mononitrate derivatives for the treatment of intestinal disorders
US9802962B2 (en) Tricyclic pyrone compounds reduce amyloid beta aggregates
KR20070026901A (en) An anticancer composition comprising obovatol or obovatal
WO2011008052A2 (en) Composition for the prevention or treatment of bone diseases comprising colforsin daropate
JP5852653B2 (en) Drug for prevention or treatment of degenerative nervous system brain disease
US20130338116A1 (en) Compounds and methods for the treatment of pain and other diseases
CA2911307C (en) Use of fluorinated derivatives of 4-aminopyridine in therapeutics and medical imaging
EP4166140A1 (en) Prophylactic or therapeutic agent for porphyria
EP3750904A1 (en) Therapeutic drug for neurodegenerative disease and application thereof
KR20220037067A (en) Pharmaceutical composition for the treatment or prevention of cerebrovascular disease
WO2020204605A1 (en) Cp2c-targeting peptide-based anticancer agent
CN115073463B (en) Matrine type dimer alkaloid compound, pharmaceutical composition and application thereof
US20240059725A1 (en) Coumarin compounds and a process for preparation thereof
WO2023050037A1 (en) Extracellular cyclophilin inhibitor and use thereof
US10766922B2 (en) Substituted steroids for the treatment of cancer
WO2021187314A1 (en) Mitochondrial dysfunction improving agent
KR20240012958A (en) Composition for anti-obesity comprising HDAC inhibitor
KR20190039271A (en) Neurodegenerative disease
US20230405134A1 (en) Novel protac chimeric compound, and pharmaceutical composition comprising same for preventing, ameliorating, or treating diseases through target protein degradation
KR100979921B1 (en) Stereum ostrea extracts, lactone compounds isolated therefrom and antiobesity composition comprising the same
US10058618B2 (en) PAK1-blocking 1,2,3-triazolyl esters

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant