WO2011091692A1 - Uses of benzoate and its derivatives - Google Patents

Abstract

The uses of benzoate and its derivatives for anti-aging of brain, and for preventing and treating the neurodegenerative diseases such as the senile dementia and the like are provided in the present invention. A series of benzoate and its derivatives are synthesized through a chemical method in the present invention. Proved through cell activity experiments in vitro, the synthesized benzoate and its derivatives have distinguished activities similar to nerve growth factors. Proved through an animal experiment, benzoate and its derivatives do not induce toxic reaction during long term oral or celiac administration, they are able to cross the blood-brain barrier and promote the regeneration of cerebral neuron.

Description

 Application of benzoic acid esters and their derivatives

Technical field

 The present invention is in the field of pharmaceuticals and relates to the use of benzoic acid esters and derivatives thereof in neurological diseases and anti-cerebral aging diseases.

Background technique

 With the aging of the social population, the prevalence of neurodegenerative diseases, especially Alzheimer's disease, has increased significantly, which has become the fourth leading cause of adult death. In particular, China's population is aging. At present, the number of patients with Alzheimer's disease, including Alzheimer's disease and vascular dementia, has exceeded 5 million, accounting for about 1/4 of the world's total cases. The lack of effective preventive and therapeutic measures has made Alzheimer's disease a serious impact on social stability and development. Therefore, the development of effective new drugs for the prevention and treatment of neurodegenerative diseases such as senile dementia is a medical problem that is urgently needed in the world. Book

 At present, the main target for clinical treatment of Alzheimer's disease drugs is the cholinergic nervous system, such as acetylcholine and acetylcholinesterase inhibitors: tacrine, rivastigmine tartrate, and huperzine A), donepezil (donepezil) and the like. However, these drugs can only partially replace the cholinergic nervous system function, temporarily improving cognitive function, and can not prevent and delay the progression of neurodegenerative diseases. Moreover, the long-term use of the effect is gradually reduced, with side effects. Therefore, the development of new drugs against brain aging and prevention of neurodegenerative diseases has become the focus of current research.

Neuronal cell death and decreased neurological function in the brain are the main causes of cognitive dysfunction. Recent studies have shown that newborn neurons (neural regeneration) in adult brain can replace and repair neuronal loss due to natural aging or lesions, and play a key role in maintaining the structure and function of the brain. Nerve growth factor (NGF) is the first biologically active peptide that has important regulatory effects on the growth, development, differentiation and function maintenance of nerves. It is the most important neurotrophic factor. Especially in the process of nervous system diseases, neurotrophic factors have important protective effects on nerve cells and nerve regeneration. Neurotrophic factors can prevent or reduce nerve atrophy, neurodegeneration, and promote nerve repair after trauma. However, it is a protein composed of more than 100 amino acids; due to its large molecular weight and strong polarity, it cannot pass the blood-brain barrier, which is a bottleneck restricting the clinical application of nerve growth factor. Currently, there is no direct administration other than intracerebral surgery. Find a better treatment. Therefore, the search for neurotrophic factor mimics (NGF mimi _CS ), NGF enhancer and NGF inducer is considered to be a new target for the prevention and treatment of neurodegenerative diseases.

 Chinese herbal medicine is the material basis of traditional Chinese medicine, and it is a cornucopia of natural active organic compounds. China has thousands of years of Chinese medicine civilization, and it has a lot of valuable local medicine. Because the utilization of Chinese herbal medicine resources is convenient, it is very important to study its new ingredients and new activities. Studies have shown that Chinese medicine has a significant effect on Alzheimer's disease, the most famous of which is the alkaloid-huperzine A extracted from Huperzia serrata. It is an anti-independent intellectual property developed by the Institute of Materia Medica, Chinese Academy of Sciences. Drugs for senile dementia. It mainly inhibits acetylcholinesterase activity, reduces glutamate-induced neuronal cell death, and has anti-beta-like peptide neurotoxicity and antioxidant activity.

Gentian, alias: bitter grass, gallbladder. Excavated in the spring and autumn seasons, washed and dried. Sexual taste, bitterness, cold. Liver, gallbladder Jing. Introduced in the 2005 edition of the Pharmacopoeia, the function and function of gentian: heat and dampness, diarrhea and biliary fire. For damp heat jaundice, vaginal itching, under the belt, strong, eczema itching, red eyes, deafness, hypochondriac pain, mouth pain, convulsions. Using the biological activity identification system established by PC12 cells, more than ten kinds of 2,3-dihydroxybenzoic acid with significant nerve growth factor activity were isolated and purified from the extracts of dried roots and rhizomes of Gmtiana rigescens Franch. A new ester compound, named gentisides o. Based on this, a series of benzoates and their derivatives (see Example 1) were synthesized. It was found that benzoate and its derivatives have similar nerve growth factor. Growth effect. In particular, the low concentration treatment of bismuth 2,3-dihydroxybenzoate (designated ABG-001) showed significant nerve growth promoting effects (see Example 2-1 for details). To date, no parabens have been reported to have similar NGF activity against Alzheimer's disease.

Summary of the invention

 The object of the present invention is to provide a preparation method and application of a benzoic acid ester and a derivative thereof in view of the deficiencies of the prior art. The object of the present invention is achieved by the following technical solutions: a benzoate and a derivative thereof for use in the preparation of a medicament for preventing and treating neurodegenerative diseases and anti-cerebral aging, the benzoate and The derivative has the following structural formula:

Wherein R is a linear, branched, saturated and unsaturated alkyl group having from 1 to 30 carbon atoms, an aliphatic ring or a derivative thereof; and R ₅ are each independently selected from the group consisting of hydrogen, a hydroxyl group, a carboxyl group, and an aldehyde group. Ester, fluorine, chlorine, bromine, iodine, sulfhydryl, amino, amide, cyano, nitro, sulfonate, trifluoromethyl, propenyl, alkyl, alkoxy, substituted benzyl, substituted phenyl , aryl, heteroaryl, glycosyl and amino acid residues; X is selected from C(0)0, OC(0), C(0)NH, NHC(0), C(0), CH ₂ , S卩0.

 Further, R is a linear, branched, saturated and unsaturated alkyl group having from 6 to 22 carbon atoms, an aliphatic ring or a derivative thereof.

Further, the benzoate and its derivative are: ethyl 2, 3-dihydroxybenzoate, amyl 2, 3-dihydroxybenzoate, octyl 2, 3-dihydroxybenzoate, 2, Ethyl 3-dihydroxybenzoate, dodecyl 2,3-dihydroxybenzoate, tetradecyl 2,3-dihydroxybenzoate, cetyl 2,3-dihydroxybenzoate, 2, Octadecyl 3-dihydroxybenzoate, eicosanyl 2,3-dihydroxybenzoate, behenyl 2,3-dihydroxybenzoate, triacontyl 2,3-dihydroxybenzoate , tetradecyl 2,6-dihydroxybenzoate, eicosanyl 2,6-dihydroxybenzoate, tetradecyl 2,4-dihydroxybenzoate, 4,4-dihydroxybenzoic acid Alkyl ester, tetradecyl 3,4,5-trihydroxybenzoate, tetradecyl 2-hydroxybenzoate, tetradecyl 3-hydroxybenzoate, 1,4-dimethoxybenzoic acid Alkyl ester, octadecyl 2, 3-dimethoxybenzoate, tetradecyl 2-hydroxy-3-methoxybenzoate, tetradecyl 2-chlorobenzoate, 2, 3-dihydroxy -N-tetradecylbenzamide, 2,3-dihydroxy-N-octylbenzene Amide, 2, 3-dihydroxy-N-dodecylbenzamide, 2,3-dihydroxy-N-hexadecylbenzamide, 2,3-dihydroxy-N-octadecylbenzene Formamide, 2,3-dimethoxy-N-tetradecylbenzamide, 3,4-dihydroxy-N-tetradecylbenzamide, 3,4,5-trihydroxy-N- Tetradecylbenzamide, N-(3,4-dimethoxyphenyl)tetradecaneamide, N- (3, 4- Dihydroxyphenyl)tetradecylidene, 1-(3,4-dimethoxyphenyl)tetradecane-1-one, 1-(3,4-dihydroxyphenyl)tetradecane- 1-ketone, 3-(tetradecyloxycarbonyl)-1,2-phenyldiacetate, 3-(tetradecylformyl)-1,2-phenyldiacetate, 5- (tetradecyloxycarbonyl)benzene-1,2,3-triacetate, 5-(tetradecylformyl)benzene-1, 2,3-triacetate, 1,2-phenylene Pericododecanoate, 2,3-dihydroxyphenyltetradecanoate, 3-(tetradecyloxy)benzene-1,2-diol.

 The beneficial effects of the invention are as follows: (1) 2,3-dihydroxybenzoate compound (ABG-001) has no toxic effect in oral or intraperitoneal administration, and the compound can rapidly enter the brain through the blood-brain barrier. Regulating the function of the cranial nervous system; (2) The results of animal pharmacology experiments prove that the 2,3-dihydroxybenzoate compound of the present invention can promote the regeneration of neurons in the adult brain, anti-brain aging, and prevent β-amyloid polypeptide The neurotoxicity, reduction of acetylcholinesterase activity, has the effect of preventing and treating senile dementia and anti-brain aging.

DRAWINGS

 Figure 1 is a dose-dependent relationship diagram of compound ABG-001 promoting neurite elongation in PC12 cells;

Figure 2 is a photomicrograph of neurites of PC 12 cells after 48 hours of addition of ABG-001, where a is 1% DMSO as a negative control; b is NGF 40 ng/ml as a positive control; c is ABG- 001 (1 μΜ) _;

 Figure 3 shows that ABG-001 has the role of a nerve growth factor, (a) is the expression of hippocampal nerve growth factor, and (b) is the number of newborn neurons;

 Figure 4 shows that ABG-001 treatment promotes nerve regeneration and migration after cerebral ischemia;

 Figure 5 shows that ABG-001 can pass the blood-brain barrier;

 Figure 6 shows that ABG-001 promotes adult nerve regeneration through the blood-brain barrier, where (a) is the number of newborn neurons, (b) is the length of the new neurons, and (c) is the differentiation of the new neurons. ;

 Figure 7 shows that ABG-001 has anti-brain aging effects, in which (a) is the number of newborn neurons, and (b), (c) and (d) are Morris water maze test plots, (e), (0 , (g) and (h) are the biochemical indicators of oxidative stress in mice; Figure 8 shows that ABG-001 has anti-cerebral aging effects, wherein (a) is the total number of times of the Y-maze,

 (b) alternate arm rates for the Y maze, and (c) and (d) are the discriminating power maps for the novel objects;

 Figure 9 shows the Morris water maze test after ABG-001 treatment for Αβ damage and Αβ damage;

 Figure 10 shows that ABG-001 treatment can improve nerve regeneration in Αβ lesions (APP/PS1 mice), where (a) is the number of newborn neurons, (b) is the length of the neonatal nerve, and (c) Acetylcholinesterase (AChE) activity map. detailed description

 2,3-dihydroxybenzoate compounds can cause a high proportion of PC12 cells to undergo neurite elongation, indicating that 2,3-dihydroxybenzoate has a very similar NGF activity and has been developed to prevent senile dementia. The value of the therapeutic drug. A series of benzoate derivatives were designed and synthesized using 2,3-dihydroxybenzoate as a lead, and their in vitro activity studies were extensively carried out to find the structure-activity relationship of the substances. It would be important to find a compound with potentially superior activity and/or lower toxicity and to prevent and treat neurodegenerative and anti-cerebral aging diseases such as Alzheimer's disease.

The benzoate derivative of the present invention, the structure of which can be represented by the formula I:

I

Wherein: R is a linear, or branched, or saturated and unsaturated alkyl group having from 1 to 30 carbon atoms, or an aliphatic ring, or a derivative thereof, especially C _{6 to} C _{22 ;}

〜 R ₅ are each independently selected from the group consisting of hydrogen, hydroxy, carboxy, aldehyde, ester, fluoro, chloro, bromo, iodo, decyl, amino, amide, cyano, nitro, sulfonyl, trifluoromethyl, Propenyl, alkyl, alkoxy, substituted benzyl, substituted phenyl, aryl, heteroaryl, glycosyl and amino acid residues;

X is selected from C (0) 0, OC ( 0), C (0) NH, NHC (0), C (0), CH 2, S and 0.

The preparation method of the benzoic acid ester and the derivative thereof of the invention is achieved by the following steps:

1) When X is c(o)o or oc(o) ester compound preparation method: first dissolve the acid with alcohol or phenol completely with solvent, cold to 0 °C, add dehydrating agent under stirring, and then rise The reaction was allowed to reach room temperature for 1 to 2 days, and the reaction was followed by thin layer chromatography. After completion of the reaction, the solvent was distilled off, worked up, and then purified by silica gel column chromatography to give an ester compound. The acid is a substituted benzoic acid or a linear, or branched, or saturated and unsaturated alkyl group having 1 to 30 carbon atoms, or an aliphatic ring, or a derivative thereof, particularly a C ₆ -C ₂₂ fatty acid; a linear or branched chain having from 1 to 30 carbon atoms, or a saturated and unsaturated alkyl group, or an aliphatic ring, or a derivative thereof, particularly a C _{6 to} C ₂₂ fatty alcohol; the phenol is a substituted phenol; The dehydrating agent is concentrated sulfuric acid, diisopropylcarbodiimide or dicyclohexylcarbodiimide; the solvent is a protic solvent methanol, ethanol or tetrahydrofuran, or an aprotic solvent dichloromethane, chloroform, benzene, toluene, two Toluene, dimethyl sulfoxide or acetonitrile; the molar ratio of acid to alcohol is 1:1 to 1:20, and the molar ratio of acid to dehydrating agent is 1:0.2~1:3.

 2) Preparation method of amide compound when X is C(0)NH or NHC(O): The acid, amine, and hydroxybenzotriazole are dissolved in a dry solvent, cooled to 0 ° C, and condensed under stirring. Agent, react at room temperature for 2 to 5 days. After the completion of the reaction, the solvent was evaporated to dryness crystals eluted eluted eluted eluted eluted eluted The acid is a substituted benzoic acid or a linear or branched, saturated or unsaturated fatty acid having from 1 to 30 carbon atoms; the amine is a substituted aniline, or a linear or branched chain having from 1 to 30, saturated or not Saturated fatty amine. The molar ratio of acid to amine is 1:1 to 1:5, and the molar ratio of acid to condensing agent is 1:1 to 1:5.

3) Preparation method of ketone compound when X is C(O): Under the protection of an inert gas, activated magnesium metal is added to anhydrous diethyl ether, and a solution of a bromoalkane in diethyl ether is added dropwise. After the start of the format reaction, the dropwise addition of a bromoalkane solution in diethyl ether was continued, and the reaction was refluxed for 1 hour. Subsequently, the prepared format reagent was added dropwise to an aldehyde solution in diethyl ether under an inert gas atmosphere at 0 ° C, and the mixture was allowed to react to room temperature for 2 hours. After the reaction is completed, the reaction solution is cooled to 0 ° C, 1 mol of hydrochloric acid per liter of hydrochloric acid solution is slowly added, and the reaction solution is extracted with diethyl ether. The organic phase is washed successively with 1 mol of diluted hydrochloric acid, saturated sodium hydrogencarbonate solution and saturated brine. Drying with anhydrous magnesium sulfate, concentrating, and purifying by silica gel column chromatography to obtain an alcohol compound. The inert gas is nitrogen or argon; the solvent is anhydrous diethyl ether or tetrahydrofuran; the aldehyde is a substituted benzaldehyde; the brominated alkane is a linear or branched, saturated or unsaturated brominated alkane having from 1 to 30; The temperature can be from -80 to 50 degrees; the molar ratio of brominated alkane to magnesium is 1:1 to 1:10; The molar ratio of the brominated alkane to the substituted benzaldehyde is from 1:1 to 1:10.

 The alcohol compound is completely dissolved in a solvent, and the oxidizing agent is slowly added. After the reaction is completed, the mixture is concentrated under reduced pressure. The obtained concentrate is diluted with ethyl acetate, and washed with 1 mol of diluted hydrochloric acid, saturated sodium hydrogen carbonate solution and saturated brine. Drying with anhydrous magnesium sulfate, concentrating, and purifying by silica gel column chromatography to obtain a ketone compound. The solvent may be tetrahydrofuran, dichloromethane, acetone, chloroform, dimethyl sulfoxide or acetonitrile; the oxidizing agent may be chromium trioxide, manganese dioxide, dimethyl sulfoxide, periodate or N-methylmorpholine oxide. The molar ratio of alcohol to oxidant is from 1:1 to 1:10.

4) Preparation method of ether compound with X being 0: The phenol is dissolved in dry tetrahydrofuran under the protection of inert gas, and alcohol, triphenylphosphine and diethyl azodicarboxylate are added under stirring, and reacted at room temperature. 1 to 24 hours. After the reaction was completed, the solvent was evaporated under reduced pressure. The concentrate was diluted with ethyl acetate, washed with brine and brine, dried over anhydrous magnesium sulfate, and evaporated. The phenol is a substituted phenol, a substituted naphthol; the alcohol is a linear or branched, saturated or unsaturated fatty alcohol having from 1 to 30 carbon atoms; the azo reagent may be diethyl azodicarboxylate or azodicarboxylic acid. Dipropyl ester, diisopropyl azodicarboxylate, di-tert-butyl azodicarboxylate or dibenzyl azodicarboxylate; the phosphine reagent may be triphenylphosphine, triisopropylphosphine, tri-p-tolyl Phosphine, triethylphosphine, tributylphosphine or tricyclohexylphosphine; solvent is protic solvent methanol or ethanol, or aprotic solvent tetrahydrofuran, dichloromethane, chloroform, benzene, toluene, xylene, dimethyl sulfoxide Or acetonitrile; the molar ratio of phenol to alcohol is 1:20, the molar ratio of phenol to azo reagent is 1 _: 0.2~1:20; the molar ratio of phenol to phosphine reagent is 1:0.2~1:20.

 The object of the present invention is the use of benzoic acid esters and derivatives thereof for the preparation of a medicament for the treatment of Alzheimer's disease and anti-cerebral aging diseases.

 The present invention further provides a pharmaceutical composition for treating senile dementia and anti-brain aging, the pharmaceutical composition comprising a physiologically effective amount of a benzoate and a derivative thereof (I) and a pharmaceutically acceptable carrier or Thinner. The weight ratio of the benzoate and its derivative (I) shown in the drug is from 0.1% to 90% by weight.

 The pharmaceutically acceptable carrier as used herein refers to a conventional pharmaceutical carrier in the pharmaceutical field, such as a diluent, an excipient, etc., a filler such as starch, sucrose, microcrystalline cellulose, etc.; a binder such as starch slurry, hydroxy Propylene, gelatin, polyethylene glycol, etc.; wetting agents such as magnesium stearate, micronized silica gel, polyethylene glycol, etc.; absorption enhancer polysorbate, lecithin, etc., surfactant poloxamer, fatty acid Yamanite, polysorbate, etc., and other adjuvants such as flavoring agents, sweeteners and the like may also be added to the composition.

 The benzoate derivatives of the present invention can be administered in unit dosage form for enteral and parenteral administration, including oral, intramuscular, and subcutaneous. The route of administration of the compound can also be intravenous. Injections include intravenous, intramuscular, subcutaneous and acupoint injections.

 The various dosage forms of the pharmaceutical compositions of the present invention can be prepared according to conventional methods of manufacture in the pharmaceutical arts, e.g., by mixing the active ingredient with one or more carriers, and then bringing it into such preparation.

 The dosage form can be a solid preparation, a capsule or a liquid preparation, including tablets, capsules, dispersible tablets, oral liquids, large infusions, small needles, freeze-dried powder needles.

The benzoate and its derivative of the present invention have remarkable nerve growth factor-like activity, have neuroprotective and anti-cerebral aging effects, and can be obtained in the prevention of neurodegenerative diseases such as Alzheimer's disease and anti-brain aging. application. The benzoate compound of the present invention exhibits remarkable NGF in the in vitro screening model PC12 cells of Alzheimer's disease Mimics activity. It is of great practical significance to use such compounds as a lead to optimize the structure and to conduct basic research for the development of new drugs for the prevention and treatment of neurodegenerative diseases such as Alzheimer's disease.

 The benzoate and its derivative of the present invention can pass the blood-brain barrier and have high-efficiency neurotrophic effects. The benzoic acid esters and derivatives thereof according to the present invention have anti-brain aging and the effects of preventing and treating senile dementia. The above-mentioned contents of the present invention will be further described in detail below by way of embodiments and drawings in which a plurality of specific compounds are prepared, but the scope of the above-mentioned subject matter of the present invention should not be construed as being limited to the following examples. The technology implemented by the above contents of the present invention is within the scope of the present invention.

Example 1

Compound 1 -1 : 2,3-dihydroxybenzoic acid ethyl ester

(154 mg, l mmol) 2,3-dihydroxybenzoic acid, 10 ml of ethanol was placed in a 25 ml round bottom flask, cooled to 0 ° C, 2 to 3 drops of concentrated sulfuric acid were added dropwise, and stirred under reflux for 24 h. The reaction was followed by thin layer chromatography (developing solvent: n-hexane/ethyl acetate, 5/1, V/V). After the reaction was stopped, ethanol was evaporated to give 390 mg of crude product. /ethyl acetate, 5/1, V/V) gave a white solid, 180 mg. 1H NMR (500 MHz, CDC1 ₃ ) δ: 11.00 (s, 1H, benzene 2-OH), 7.38 (dd, 1 Η, / = 1.5, 8.0 Hz, benzene H-6), 7.12 (dd, 1H, / = 1.0, 7.5 Hz, benzene H-4), 6.80 (t, 1H, J = 8.0 Hz, benzene H-5), 5.66 (s, 1H, benzene 3-OH), 4.41 (q, 2H, J = 7.0 Hz ), 1.42 (t, 3H, J = 7.0 Hz); HRMS: m/z [M+H]+ calcd for C ₉ H _u 0 ₄ ⁺ : 183.0652, found: 183.0639.

Compound I -2: Amyl 2,3-dihydroxybenzoate

The synthesis was carried out in the same manner as Compound 1-1, and the reaction was carried out as follows: (154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, 10 ml of pentanol, to give a white solid, 170 mg o 1H NMR (500 MHz, CDC1 ₃ ) δ: 11.00 (s, 1H), 7.38 (dd, 1H, J = 1.5, 8.0 Hz), 7.12 (dd, 1H, / = 1.0, 8.0 Hz), 6.80 (t, 1H, / = 8.0 Hz), 5.65 (s , 1H), 4.41 (t, 2H, / = 7.0 Hz), 1.78 (m, 2H), 1.33~ 1.38 (m, 4H), 0.90 (t, 3H, / = 7.0 Hz); ¹³ C NMR (125 MHz , CDC1 ₃ ) δ: 170.4, 148.9, 145.1, 120.5, 119.7, 119.1, 112.7, 65.7, 28.2, 28.1, 22.3, 13.9 ppm; IR (KBr) v: 3458, 2931, 2865, 1674, 1469, 1309, 1268 , 1067, 754 cm ¹ ; HRMS: m/z [M+H]+ calcd for C ₁₂ H ₁₇ 0 ₄ ⁺ : 225.1121 , found: 225.1104.

Compound I -3 : 2,3-dihydroxybenzoic acid octyl ester

The synthesis was carried out in the same manner as Compound 1-1, and the reaction was carried out as follows: (154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, 10 ml of octanol, obtained as a white solid, 128 mg o 1H NMR (500 MHz, CDC1 ₃ ) δ: 11.01 (s, 1Η), 7.38 (dd, 1Η, / = 1.5, 8.0 Hz), 7.10 (dd, 1H, / = 1.0, 8.0 Hz), 6.80 (t, 1H, / = 8.0 Hz), 5.64 (s , 1H), 4.35 (t, 2H, / = 7.0 Hz), 1.78 (m, 2H), 1.44 (m, 2H), 1.26~ 1.38 (m, 8H), 0.89 (t, 3H, / = 7.0 Hz) ; HRMS: m/z [M+H]+ calcd for C ₁₅ H ₂₃ 0 ₄ ⁺ : 267.1591 , found: 267.1590.

Compound I -4: 2,3-dihydroxybenzoate decyl ester

(154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (316 mg, 2 mmol) n-nonanol, 10 ml tetrahydrofuran in a 25 ml round bottom flask, cooled to 0 ° C, added (250 mg , 1.2 mmol) Dicyclohexylcarbodiimide, stirred at room temperature for 24 h. The reaction was followed by thin layer chromatography (developing solvent: n-hexane/ethyl acetate, 2/1, V/V). After the reaction was stopped, the solvent was evaporated, the residue was evaporated, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated. Primary product 440 mg, silica gel column chromatography (developing solvent: n-hexane/ethyl acetate, 2/1, V/V) gave a white solid 132 mg. 1HNMR (500 MHz, CDC1 ₃ ) δ: 11.01 (s, 1H), 7.37 (dd, 1Η, /= 1.5, 8.0 Hz), 7.09 (dd, 1H, /= 1.5, 8.0 Hz), 6.80 (t, 1H , /= 8.0 Hz), 5.65 (s, 1H), 4.35 (t, 2H, /= 7.0 Hz), 1.78 (m, 2H), 1.44 (m, 2H), 1.27~ 1.35 (m, 12H), 0.88 (t, 3H, / = 7.0 Hz); ¹³ C NMR (125 MHz, CDCI3) δ: 170.8, 148.9, 145.0, 120.5, 120.0, 119.6, 119.1, 65.7, 31.9, 29.5, 29.3, 29.2, 28.5, 27.6, 25.9, 22.7, 14.1 ppm; IR (KBr) v: 3478, 2926, 2885, 1667, 1469, 1309, 1267, 1067, 753 cm"¹; HRMS: m/z [M+H]+ calcd for C ₁₇ H ₂₇ 0 ₄ ⁺ : 295.1904, found: 295.1910.

Compound I -5: Dodecyl 2,3-dihydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: (154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (372 mg, 2 mmol), dodecanol, (250 mg, 1.2 mmol) dicyclohexyl A carbodiimide, 15 ml of tetrahydrofuran afforded a white solid, 180 mg. 1H NMR (500 MHz, CDC1 ₃ ) δ: 11.01 (s, 1Η), 7.38 (dd, 1Η, /= 1.5, 8.0 Hz), 7.10 (dd, 1Η, /= 1.0, 8.0 Hz), 6.80 (t, 1H, /= 8.0 Hz), 5.64 (s, 1H), 4.35 (t, 2H, /= 6.5 Hz), 1.78 (m, 2H), 1.44 (m, 2H), 1.26~ 1.35 (m, 16H), 0.88 (t, 3H, /= 7.0 Hz); HRMS: m/z [M+H]+ calcd for C ₁₉ H ₃₁ 0 ₄ ⁺ : 323.2217, found: 323.2236.

Compound I -6: tetradecyl 2,3-dihydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: ( 154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (428 mg, 2 mmol) tetradecyl alcohol, (250 mg, 1.2 mmol) dicyclohexyl A carbodiimide, 15 ml of tetrahydrofuran afforded 150 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 11.01 (s, 1Η), 7.37 (dd, 1Η, /= 1.5, 8.5 Hz), 7.10 (dd, 1Η, /= 1.0, 8.0 Hz), 6.80 (t, 1H, /= 8.0 Hz), 5.63 (s, 1H), 4.35 (t, 2H, /= 6.5 Hz), 1.78 (m, 2H), 1.44 (m, 2H), 1.26~ 1.35 (m, 20H), 0.88 (t, 3H, / = 7.0 Hz); 13 C NMR (125 MHz, CDC1 3) δ: 170.4, 148.9, 145.0, 120.5, 119.6, 119.1, 112.6, 65.7, 31.9, 29.7~ 29.6, 29.5, 29.4, 29.2, 28.5, 25.9, 22.7, 14.1 ppm; IR (KBr) v: 3485, 2917, 2849, 1669, 1467, 1310, 1267, 1157, 1067, 759 cm"¹; HRMS: m/z [M+H] + calcd for C ₂₁ H ₃₅ 0 ₄ ⁺ : 351.2530, found: 351.2536.

Compound I -7: Cetyl 2,3-dihydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: (154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (484 mg, 2 mmol) cetyl alcohol, (250 mg, 1.2 mmol) dicyclohexyl A carbodiimide, 15 ml of tetrahydrofuran afforded 178 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 11.01 (s, 1Η), 7.37 (dd, 1Η, /= 1.5, 8.0 Hz), 7.10 (dd, 1Η, /= 1.0, 8.0 Hz), 6.80 (t, 1H, /= 8.0 Hz), 5.65 (s, 1H), 4.35 (t, 2H, /= 6.5 Hz), 1.78 (m, 2H), 1.44 (m, 2H), 1.25~ 1.35 (m, 24H), 0.88 (t, 3H, /= 7.0 Hz); HRMS: m/z [M+H]+ calcd for C ₂₃ H ₃₉ 0 ₄ ⁺ : 379.2843, found: 379.2848.

Compound I -8: Octadecyl 2,3-dihydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: (154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (540 mg, 2 mmol) octadecyl alcohol, (250 mg, 1.2 mmol) dicyclohexyl The carbodiimide, 15 ml of tetrahydrofuran afforded 170 mg of a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 11.01 (s, 1Η), 7.37 (dd, 1Η, /= 1.5, 8.0 Hz), 7.10 (dd, 1Η, /= 1.0, 8.0 Hz), 6.80 (t, 1H, /= 8.0 Hz), 5.63 (s, 1H), 4.35 (t, 2H, /= 6.5 Hz), 1.78 (m, 2H), 1.44 (m, 2H), 1.25~ 1.35 (m, 28H), 0.88 (t, 3H, /= 7.0 Hz); ¹³ C NMR (125 MHz, CDC1 ₃ ) δ: 170.4, 148.9, 145.1, 120.5, 119.6, 119.1, 112.7, 65.7, 31.9, 29.7~ 29.6, 29.5, 29.4, 29.2, 28.5, 25.9, 22.7, 14.1 ppm; IR (KBr) v: 3473, 2917, 2850, 1665, 1467, 1266, 1159, 1064, 799, 762 cm- ¹ ; HRMS: mJz [M+H]+ calcd for C ₂₅ H ₄₃ 0 ₄ ⁺ : 407.3156, found : 407.3192.

Compound I -9: Eicosanyl 2,3-dihydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: ( 154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (896 mg, 3 mmol) eicosyl alcohol, (206 mg, 1 mmol) dicyclohexyl The carbodiimide, 15 ml of tetrahydrofuran afforded 104 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 11.00 (s, 1Η), 7.37 (dd, 1Η, / = 1.5, 8.0 Hz), 7.10 (dd, 1Η, / = 0.5, 8.0 Hz), 6.80 (t, 1H, / = 8.0 Hz), 5.63 (s, 1H), 4.34 (t, 2H, / = 6.5 Hz), 1.78 (m, 2H), 1.43 (m, 2H), 1.25~ 1.35 (m, 32H), 0.88 (t, 3H, / = 7.0 Hz); ¹³ C NMR (125 MHz, CDC1 ₃ ) δ: 170.4, 148.9, 145.0, 120.5, 119.6, 119.1, 112.7, 65.7, 31.9, 29.7~ 29.6, 29.5, 29.4, 29.2, 28.5, 25.9, 22.7, 14.1 ppm; IR (KBr) v: 3403, 2919, 2850, 1675, 1467, 1313, 1254, 1159, 1067, 752 cm- ¹ ; HRMS: mJz [M+H]+ calcd For C ₂₇ H ₄₇ 0 ₄ ⁺ : 435.3469, found: 435.3489.

Compound I -10: behenyl 2,3-dihydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: ( 154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (653 mg, 2 mmol) dodecanol, (250 mg, 1.2 mmol) Hexylcarbodiimide, 15 ml of tetrahydrofuran afforded 102 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 11.01 (s, 1H), 7.37 (dd, 1Η, / = 1.5, 8.0 Hz), 7.10 (dd, 1Η, / = 1.0, 8.0 Hz), 6.80 (t, 1H, / = 8.0 Hz), 5.63 (s, 1H), 4.34 (t, 2H, / = 6.5 Hz), 1.78 (m, 2H), 1.43 (m, 2H), 1.25~ 1.35 (m, 36H), 0.88 (t, 3H, / = 7.0 Hz); ¹³ C NMR (125 MHz, CDC1 ₃ ) δ: 170.4, 148.9, 145.1, 120.5, 119.6, 119.1, 112.7, 65.7, 31.9, 29.7~ 29.6, 29.5, 29.4, 29.2, 28.5, 25.9, 22.7, 14.1 ppm; IR (KBr) v: 3397, 2918, 2849, 1675, 1469, 1312, 1256, 1158, 1065, 750 cm"¹; HRMS: m/z [M+H] + calcd for C ₂₉ H ₅₁ 0 ₄ ⁺ : 463.3782, found: 463.3790.

Compound I -11 : Trialkyl ester of 2,3-dihydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: (154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, ( 1.32 g, 3 mmol) triacontanol, (250 mg, 1.2 mmol) dicyclohexyl A carbodiimide, 20 ml of tetrahydrofuran afforded a white solid, 75 mg. 1H NMR (500 MHz, CDC1 ₃ ) δ: 11.00 (s, 1H), 7.37 (dd, 1Η, / = 1.5, 8.5 Hz), 7.10 (dd, 1Η, / = 1.0, 8.0 Hz), 6.80 (t, 1H, / = 8.0 Hz), 5.63 (s, 1H), 4.34 (t, 2H, / = 6.5 Hz), 1.77 (m, 2H), 1.45 (m, 2H), 1.25~ 1.35 (m, 52H), 0.88 (t, 3H, / = 7.0 Hz); ¹³ C NMR (125 MHz, CDC1 ₃ ) δ: 164.5, 149.4, 145.0, 120.5, 119.6, 119.1, 112.6, 65.7, 34.9, 31.9, 29.7~ 29.5, 29.4, 29.2, 28.5, 25.9, 25.4, 24.7, 22.7, 14.1 ppm; IR (KBr) v: 3484, 2920, 2848, 1665, 1468, 1313, 1262, 1162, 1076, 801 cm- ¹ ; MS (mz): 575 [M] ⁺ .

Compound I -12: tetradecyl 2,6-dihydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: ( 154 mg, 1 mmol) 2,6-dihydroxybenzoic acid, (428 mg, 2 mmol) tetradecyl alcohol, (250 mg, 1.2 mmol) dicyclohexyl A carbodiimide, 15 ml of tetrahydrofuran afforded a white solid, 66 mg. 1H NMR (500 MHz, CDC1 ₃ ) δ: 9.79 (s, 2H), 7.32 (t, 1H, J = 8.5 Hz), 6.48 (d, 2H, J = 8.5 Hz), 4.35 (t, 2H, / = 6.5 Hz), 1.78 (m, 2H), 1.44 (m, 2H), 1.26~ 1.35 (m, 20H), 0.88 (t, 3H, / = 7.0 Hz); ¹³ C NMR (125 MHz, CDC1 ₃ ) δ : 169.8, 149.3, 145.4, 160.9, 136.5, 108.2, 100.1, 66.7, 53.4, 31.9, 29.6 to 29.7, 29.5, 29.4, 29.3, 29.1 , 28.5, 25.9, 22.7, 14.1 ppm; IR (KBr) v: 3472, 2919, 2849, 1669, 1630, 1576, 1465, 1327, 1293, 1156, 1067, 782 cm"¹; HRMS: m/z [M+H]+ calcd for C ₂₁ H ₃₅ 0 ₄ ⁺ : 351.2530, found: 351.2516.

Compound I -13 : Eicosanyl 2,6-dihydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: ( 154 mg, 1 mmol) 2,6-dihydroxybenzoic acid, (597 mg, 2 mmol) eicosyl alcohol, (250 mg, 1.2 mmol) dicyclohexyl The carbodiimide, 15 ml of tetrahydrofuran afforded 104 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 9.79 (s, 2Η), 7.31 (t, 1H, / = 8.5 Hz), 6.48 (d, 2H, / = 8.5 Hz), 4.50 (t, 2H, / = 6.5 Hz), 1.83 (m, 2H), 1.43 (m, 2H), 1.25~ 1.35 (m, 32H), 0.88 (t, 3H, / = 7.0 Hz); MS (m/z): 434 [M] ⁺ .

Compound I -14: tetradecyl 2,4-dihydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: ( 154 mg, 1 mmol) 2,4-dihydroxybenzoic acid, (428 mg, 2 mmol) tetradecyl alcohol, (250 mg, 1.2 mmol) dicyclohexyl A carbodiimide, 15 ml of tetrahydrofuran afforded 158 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 11.06 (s, 1H), 7.74 (d, / = 8.5 Hz, 1H), 7.54 (m, 1H), 6.38 (m, 1H), 5.39 (s, 1H) , 4.30 (t, / = 6.5 Hz, 2H), 1.76 (m, 2H), 1.41 (m, 2H), 1.25~ 1.35 (m, 20H), 0.88 (t, / = 7.0 Hz, 3H) ppm; HRMS : m/z [M+H]+ calcd for C ₂₁ H ₃₅ 0 ₄ ⁺ : 351.2530, found: 351.2557.

Compound I -15 : tetradecyl 3,4-dihydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: ( 154 mg, 1 mmol) 2,4-dihydroxybenzoic acid, (428 mg, 2 mmol) tetradecyl alcohol, (250 mg, 1.2 mmol) dicyclohexyl A carbodiimide, 15 ml of tetrahydrofuran afforded a white solid, 180 mg. 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.59 (d, J = 2.0 Hz, 1H), 7.57 (dd, J = 2.0, 8.0 Hz, 1H), 6.91 (d, J = 8.5 Hz, 1H), 4.26 (t, / = 6.5 Hz, 2H), 1.73 (m, 2H), 1.42 (m, 2H), 1.25~ 1.37 (m, 20H), 0.89 (t, / = 7.0 Hz, 3H) ppm; HRMS: m /z [M+H]+ calcd for C ₂₁ H ₃₅ 0 ₄ ⁺ : 351.2530, found: 351.2551.

Compound I -16: tetradecyl 3,4,5-trihydroxybenzoate

The synthesis method is the same as the compound I -4, and the reaction charge is: (170 mg, 1 mmol) 3,4,5-trihydroxybenzoic acid, (428 mg, 2 mmol) tetradecanol, (250 mg, 1.2 mmol) Cyclohexylcarbodiimide, 15 ml of tetrahydrofuran afforded 176 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.24 (s, 2H), 4.24 (t, / = 6.5 Hz, 2H), 1.72 (m, 2H), 1.41 (m, 2H), 1.25~ 1.35 (m, 20H), 0.88 (t, J = 7.0 Hz, 3H) ppm; HRMS: m/z [M+H]+ calcd for C ₂₁ H ₃₅ 0 ₅ ⁺ : 367.2479, found: 367.2473.

Compound I -17 : tetradecyl 2-hydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: ( 138 mg, 1 mmol) 2-hydroxybenzoic acid, (428 mg, 2 mmol) tetradecyl alcohol, (250 mg, 1.2 mmol) dicyclohexylcarbazide Amine, 15 ml of tetrahydrofuran afforded 164 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 10.88 (s, 1H), 7.37 (m, 1H), 7.10 (m, 1H), 6.91 (m, 1H), 6.80 (t, 1H, J = 8.0 Hz) , 4.35 (t, 2H, / = 6.5 Hz), 1.78 (m, 2H), 1.44 (m, 2H), 1.26~ 1.35 (m, 20H), 0.88 (t, 3H, / = 7.0 Hz); HRMS: m/z [M+H]+ calcd for C ₂₁ H ₃₅ 0 ₃ ⁺ : 335.2581 , found: 335.2564.

Compound I -18 : tetradecyl 3-hydroxybenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: ( 138 mg, 1 mmol) 3-hydroxybenzoic acid, (428 mg, 2 mmol) tetradecyl alcohol, (250 mg, 1.2 mmol) dicyclohexylcarbazide Amine, 15 ml of tetrahydrofuran afforded 174 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.57 (m, 1H), 7.37 (m, 1H), 7.25 (m, 1H), 7.01 (m, 1H), 5. 67 (s, 1H), 4.35 (t, 2H, / = 6.5 Hz), 1.78 (m, 2H), 1.44 (m, 2H), 1.26~ 1.35 (m, 20H), 0.88 (t, 3H, / = 7.0

Hz); MS (m/z): 334 [M] ⁺ .

Compound I -19: tetradecyl 2,3-dimethoxybenzoate

The synthesis method is the same as the compound I -4, and the reaction charge is: (182 mg, l mmol) 2,3-dimethoxybenzoic acid, (428 mg, 2 mmol) tetradecanol, (250 mg, 1.2 mmol) Cyclohexylcarbodiimide, 15 ml of tetrahydrofuran afforded 162 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.08 (t, 1 Η, / = 7.5 Hz), 6.95 (dd, 1H, / = 1.5, 8.0 Hz), 6.79 (dd, 1H _: / = 1.0, 7.0 Hz) , 4.35 (t, 2H, J = 6.5 Hz), 3.92 (s, 3H), 3.88 (s, 3H), 1.78 (m, 2H), 1.44 (m, 2H), 1.26~ 1.35 (m, 20H), 0.88 (t, 3H, / = 7.0 Hz); ¹³ C NMR (125 MHz, CDC1 ₃ ) δ: 166.4, 153.5, 148.9, 126.6, 123.8, 122.2, 115.6, 65.3, 61.5, 56.0, 31.9, 29.7~ 29.6, 29.5, 29.3, 29.2, 28.7, 26.0, 22.7, 14.1 ppm; IR (KBr) v: 2926, 2855, 1726, 1587, 1474, 1266, 1148, 1062, 754 cm"¹; HRMS: m/z [ +H ] ⁺ calcd for C ₂₃ H ₃₉ 0 ₄ ⁺ : 379.2843, found: 379.2865.

Compound I -20: Octadecyl 2,3-dimethoxybenzoate

The synthesis method is the same as the compound I -4, and the reaction charge is: (182 mg, l mmol) 2,3-dimethoxybenzoic acid, (540 mg, 2 mmol) stearyl alcohol, (250 mg, 1.2 mmol) Cyclohexylcarbodiimide, 15 ml of tetrahydrofuran afforded a white solid, 170 mg. 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.06 (t, 1H, / = 8.0 Hz), 6.93 (dd, 1H, / = 1.5, 8.0 Hz), 6.77 (dd, 1H _: / = 1.5, 8.0 Hz) , 4.35 (t, 2H, J = 6.5 Hz), 3.92 (s, 3H), 3.88 (s, 3H), 1.78 (m, 2H), 1.44 (m, 2H), 1.25~ 1.35 (m, 28H), 0.88 (t, 3H, / = 7.0 Hz); MS (mz): 434 [M]+.

Compound I -21 : tetradecyl 2-hydroxy-3-methoxybenzoate

The synthesis method is the same as the compound I -4, and the reaction charge is: (168 mg, 1 mmol) 2-hydroxy-3-methoxybenzoic acid, 428 mg, 2 mmol) tetradecanol, (250 mg, 1.2 mmol) Cyclohexylcarbodiimide, 15 ml of tetrahydrofuran, obtained as a yellow liquid, 109 mg, 1H NMR (500 MHz, CDC1 ₃ ) δ: 11.12 (s, 1H), 7.44 (dd, / = 1.5, 8.5 Hz, 1H), 7.04 ( Dd, 1H, J = 1.5, 8.0 Hz, 1H), 6.83 (t, / = 8.0 Hz, 1H), 4.34 (t, / = 6.5 Hz, 2H), 3.91 (s, 3H), 1.77 (m, 2H) ), 1.44 (m, 2H), 1.25~ 1.36 (m, 20H), 0.88 (t, J = 7.0 Hz, 3H) ppm; ¹³ C NMR (125 MHz, CDC1 ₃ ) δ: 170.5, 152.1, 148.5, 121.0 , 118.4, 116.4, 112.9, 65.6, 56.2, 31.9, 29.7 to 29.6, 29.5, 29.3, 29.2, 28.5, 25.9, 22.7, 14.1 ppm; IR (KBr) v: 3080, 2923, 2849, 1667, 1587, 1466, 1341, 1242, 1167, 1062, 767 cm ¹ ; HRMS: m/z [M+H]+ calcd for C ₂₂ H ₃₇ 0 ₄ ⁺ : 365.2686, found: 365.2654.

Compound I -22: tetradecyl 2-chlorobenzoate

The synthesis was carried out in the same manner as the compound I -4, and the reaction was carried out as follows: ( 156 mg, 1 mmol) 2-chlorobenzoic acid, (428 mg, 2 mmol) tetradecyl alcohol, (250 mg, 1.2 mmol) dicyclohexylcarbodiimide Amine, 15 ml of tetrahydrofuran afforded 150 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.65 (m, 1H), 7.30 (m, 2H), 7.08 (m, 1H), 4.35 (t, 2H, / = 6.5 Hz), 1.78 (m, 2H) , 1.44 (m, 2H), 1.26~ 1.35 (m, 20H), 0.88 (t, 3H, / = 7.0 Hz); MS (mz): 352 [M]+.

Compound I -23 : 2,3-dihydroxy-N-tetradecylbenzamide

(154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (135 mg, 1 mmol) 1-hydroxybenzotriazole hydrate, (253 mg, 1.1 mmol) tetradecylamine, 10 ml of tetrahydrofuran In a 25 ml round bottom flask, cold at 0 ° C, (250 mg, 1.2 mmol) of dicyclohexylcarbodiimide was added and stirred at room temperature for 4 h. The reaction was followed by thin layer chromatography (developing solvent: n-hexane/ethyl acetate, 3/1, V/V). After the reaction was stopped, the solvent was evaporated, the residue was crystallised from ethyl acetate, filtered, and the filtrate was dissolved in saturated sodium hydrogen carbonate. The organic layer was dried over anhydrous sodium sulfate, filtered, and evaporated to give 440 mg of the crude product. ¹ l·^ NMR (500MHz, CDC1 ₃ ) δ: 12.81 (s, 1H), 7.04 (dd, /= 1.0, 8.0 Hz, 1H), 6.87 (dd, /= 1.0, 8.0 Hz, 1H), 6.76 ( t, /= 8.0 Hz, 1H), 6.30 (s, 1H), 5.78 (s, 1H), 3.44 (m, 2H), 1.62 (m, 2H), 1.25~ 1.40 (m, 22H), 0.88 (t , / = 7.5 Hz, 3H) ppm; ¹³ C NMR (125 MHz, CDC1 ₃ ) δ: 169.9, 149.1, 146.0, 118.5, 117.9, 115.7, 114.0, 39.7, 31.9, 29.7~ 29.6, 29.5, 29.4, 29.3, 29.2, 26.9, 22.7, 14.1 ppm; IR (KBr) v: 3482, 3389, 3268, 2920, 2850, 1639, 1586, 1546, 1462, 1338, 1270, 1236, 1176, 1077, 723, 746 cm"¹; HRMS: m/z [M+Na]+ calcd for C ₂₁ H ₃₅ N0 ₃ Na ⁺ : 372.2509, found: 372.2499. Compound I -24: 2,3-dihydroxy-N-octylbenzamide

The synthesis was carried out in the same manner as the compound I-23, and the reaction was carried out as follows: (154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (185 mg, 1 mmol) octylamine, (135 mg, 1 mmol) 1-hydroxybenzo Triazole hydrate, (250 mg, 1.2 mmol), dicyclohexylcarbodiimide, 15 ml of tetrahydrofuran, 158 mg, 1H NMR (500 MHz, CDC1 ₃ ) δ: 12.81 (s, 1 Η), 7.04 (d, / = 8.0 Hz, 1H), 6.86 (d, /= 8.5 Hz, 1H), 6.76 (t, /= 8.0 Hz, 1H), 6.31 (s, 1H), 5.78 (s, 1H), 3.44 (q, J = 7.0 Hz, 2H), 1.63 (m, 2H), 1.27~ 1.39 (m, 10H), 0.88 (t, /= 7.0 Hz, 3H); MS (m/z): 265 [M]+. I -25: 2,3-dihydroxy-N-dodecylbenzamide

Synthetic method with compound I-23, the reaction charge was: (154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (428 mg, 2 mmol) dodecylamine, (135 mg, 1 mmol) 1-hydroxyl Benzotriazole hydrate, (250 mg, 1.2 mmol) of dicyclohexylcarbodiimide, 15 ml of tetrahydrofuran afforded 158 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 12.81 (s, 1H, Ph2-OH), 7.04 (dd, /= 1.0, 8.0 Hz, 1H), 6.87 (dd, /= 1.0, 8.5 Hz, 1H), 6.75 (t, /= 8.0 Hz, 1H), 6.32 (s, 1H), 5.80 (s, 1H), 3.44 (q, /= 7.0 Hz, 2H), 1.62 (m, 2H), 1.25~ 1.40 (m , 10H), 0.88 (t, / = 7.0 Hz, 3H); MS (m/z): 321 [M] ⁺ .

Compound I -26: 2,3-dihydroxy-N-hexadecylbenzamide

Synthetic method with compound I-23, the reaction charge was: (154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (241 mg, 1 mmol) cetylamine, (135 mg, 1 mmol) 1-hydroxyl Benzotriazole hydrate, (250 mg, 1.2 mmol) of dicyclohexylcarbodiimide, 15 ml of tetrahydrofuran afforded 308 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 12.81 (s, 1H, Ph2-OH), 7.04 (dd, /= 1.0, 8.0 Hz, 1H), 6.86 (dd, /= 1.0, 8.5 Hz, 1H), 6.75 (t, /= 8.0 Hz, 1H), 6.30 (s, 1H), 5.77 (s, 1H), 3.44 (q, J= 6.5 Hz, 2H), 1.61 (m, 2H), 1.25~ 1.40 (m , 10H), 0.88 (t, / = 7.0 Hz, 3H); MS (m/z): 377 [M] ⁺ .

Compound I -27: 2,3-dihydroxy-N-octadecylbenzamide

Synthetic method with compound I-23, the reaction charge was: (154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (269 mg, 1 mmol) octadecylamine, (135 mg, 1 mmol) 1-hydroxyl Benzotriazole hydrate, (250 mg, 1.2 mmol) of dicyclohexylcarbodiimide, 15 ml of tetrahydrofuran afforded 340 mg of white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 12.81 (s, 1H, Ph2-OH), 7.04 (dd, J=1.0, 8.0 Hz, 1H), 6.87 (dd, /= 1.0, 8.0 Hz, 1H), 6.76 (t, /= 8.0 Hz, 1H), 6.30 (s, 1H), 5.77 (s, 1H), 3.44 (q, /= 6.5 Hz, 2H), 1.63 (m, 2H), 1.25~ 1.38 (m , 30H), 0.88 (t, / = 7.5 Hz, 3H); MS (m/z): 405 [M] ⁺ .

Compound I -28: 2,3-dimethoxy-N-tetradecylbenzamide

The synthesis method is the same as the compound I -23, and the reaction charge is: (182 mg, 1 mmol) 2,3-dimethoxybenzoic acid, (230 mg, l Methyl)tetradecylamine, (135 mg, 1 mmol) 1-hydroxybenzotriazole hydrate, (250 mg, 1.2 mmol) dicyclohexylcarbodiimide, 15 ml of tetrahydrofuran afforded 305 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.97 (s, 1 Η, ΝΗ), 7.68 (dd, /= 1.0, 8.0 Hz, 1 Η), 7.14 (t, /= 8.0 Hz, 1H), 7.02 (dd, / = 1.0, 8.0 Hz, 1H); MS (mz): 377 [M] ⁺ .

Compound I -29: 3,4-dihydroxy-N-tetradecylbenzamide

Synthetic method with compound I -23, the reaction charge was: (154 mg, 1 mmol) 2,3-dihydroxybenzoic acid, (230 mg, 1 mmol) tetradecylamine, (135 mg, 1 mmol) 1-hydroxyl Benzotriazole hydrate, (250 mg, 1.2 mmol) of dicyclohexylcarbodiimide, 15 ml of tetrahydrofuran afforded 297 mg as a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.66 (d, / = 2.0 Hz, 1H), 7.09 (dd, /= 2.0, 8.0 Hz, 1H), 6.88 (d, /= 8.0 Hz, 1H), 6.12 (s, 1H), 3.43 (m, 2H), 1.93 (m, 2H), 1.25~ 1.39 (m, 22H), 0.88 (t, / = 7.0 Hz, 3H) ppm; ¹³ C NMR (125 MHz, CDC1 ₃ ) δ: 167.9, 148.0, 144.1, 118.8, 115.5, 115.7, 114.0, 40.3, 31.9, 29.7 to 29.6, 29.5, 29.3, 27.0, 22.7, 14.1 ppm; IR (KBr) v: 3494, 3382, 3182, 2921 , 2851, 1587, 1517, 1467, 1439, 1294, 1169, 1106, 769 cm- ¹ ; HRMS: mJz [M+H]+ calcd for C ₂₁ H ₃₆ N0 ₃ ⁺ : 350.2690, found: 350.2683.

Compound I -30: 3,4,5-trihydroxy-N-tetradecylbenzamide

The synthesis was carried out in the same manner as the compound I-23, and the reaction was carried out as follows: (170 mg, 1 mmol) 3,4,5-trihydroxybenzoic acid, (230 mg, 1 mmol) tetradecylamine, (135 mg, 1 mmol) 1 -Hydroxybenzotriazole hydrate, (250 mg, 1.2 mmol) of dicyclohexylcarbodiimide, 15 ml of tetrahydrofuran afforded 314 mg as a white solid. 1HNMR (500 MHz, CDC1 ₃ ) S: 7.34~ 7.37 (m, 3H), 3.65 (t, /= 8.0 Hz, 2H), 2.26 (s, 3H), 2.24 (s, 3H), 1.52 (m, 2H) ), 1.21~ 1.31 (m, 22H), 0.88 (t, /= 7.0 Hz, 3H); MS (m/z): 365 [M] ⁺ .

Compound I -31: N-(3,4-dimethoxyphenyl)tetradecanoylamide

The synthesis method is the same as the compound I-23, and the reaction charge is: (312 mg, 2 mmol) 3,4-dimethoxyaniline, (465 mg, 2 mmol) myristic acid, (135 mg, 1 mmol) 1-hydroxybenzene The triazole hydrate, (450 mg, 2.2 mmol) of dicyclohexylcarbodiimide, 30 ml of tetrahydrofuran afforded 560 mg of white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.40 (d, /= 2.0 Hz, 1H), 7.05 (s, 1H), 6.82 (dd, / = 2.0, 8.5 Hz, 1H), 6.80 (s, 1H) , 3.88 (s, 3H), 3.86 (s, 3H), 2.33 (t, /= 7.5 Hz, 2H), 1.72 (m, 2H), 1.25~ 1.40 (m, 20H), 0.88 (t, J= 7.0 Hz, 3H) ppm; MS (m/z): 363 [M]+. Compound I -32: N-(3,4-dihydroxyphenyl)tetradecylidene

(150 mg, 0.41 mmol) of compound 1-31 was dissolved in 10 ml of dichloromethane, and boron tribromide (0.3 ml, 3 mmol) was added dropwise at 0 °, and the mixture was stirred at room temperature and stirred overnight. After the completion of the reaction, the mixture was evaporated to dryness eluted Silica gel column chromatography (developing solvent: petroleum ether / ethyl acetate, 8/1, V/V) as a white solid, 118 mg, 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.29 (d, / = 2.0 Hz, 1H) , 7.15 (s, 1H), 6.79 (d, /= 8.5 Hz, 1H), 6.40 (dd, /= 2.5, 8.5 Hz, 1H), 5.30 (s, 2H), 2.38 (t, / = 7.5 Hz, 2H), 1.74 (m, 2H), 1.26~ 1.40 (m, 20H), 0.88 (t, /= 7.0 Hz, 3H) ppm; MS (mz): 335 [M]+.

Compound I -33: l-(3,4-Dimethoxyphenyl)tetradecane-1-one

Tetradecanic acid (2.28 g, 0.01 mol) was dissolved in 30 ml of thionyl chloride, refluxed under nitrogen overnight, and excess SOCl ₂ was distilled off under reduced pressure to give tetradecyl chloride. 2 ml of 1,2-dimethoxybenzene was dissolved in 20 ml of carbon disulfide, and A1C1 ₃ (1.4 g, 10 mmol) was added portionwise with stirring. After the addition, the mixture was stirred for another 15 minutes, and the acid chloride was added dropwise, and stirring was continued for 4 hours. After the reaction is completed, the reaction will be Pour the solution into 30 ml of ice water until the reddish brown color disappears. Extract with ethyl acetate, dry over anhydrous sodium sulfate, filtered and evaporated. Silica gel column chromatography (developing solvent: petroleum ether / ethyl acetate, 10/1, V/V) 1H NMR (500 MHz, CDCl ₃ ) δ: 7.58 (dd, J = 2.0, 8.5 Hz, 1H), 7.53 (d, J = 2.0 Hz, 1H), 6.88 (d, J = 8.5 Hz, 1H), 6.92 (d, J = 8.5 Hz, 1H), 3.95 (s, 3H), 3.94 (s, 3H), 2.92 (t, / = 7.5 Hz, 2H), 1.72 (m, 2H), 1.25~ 1.40 (m, 20H), 0.88 (t, / = 6.5 Hz, 3H) ppm; HRMS: m/z [M+H]+ calcd for C ₂₂ H ₃₇ 0 ₃ ⁺ : 349.2737, found: 349.2715. Compound I -34: l- (3,4-dihydroxyphenyl)tetradecane-1-one

(348 mg, 1 mmol) of compound 1-33 was dissolved in 20 ml of dichloromethane, and boron tribromide (0.6 ml, 6 mmol) was added dropwise at 0 °, and the mixture was stirred at room temperature and stirred overnight. After completion of the reaction, the mixture was evaporated to dryness eluted Silica gel column chromatography (developing solvent: petroleum ether/ethyl acetate, 8/1, V/V) 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.67 (d, / = 2.0 Hz, 1H), 7.50 (dd, / = 2.0, 8.5 Hz, 1H), 6.92 (d, / = 8.5 Hz, 1H), 6.17 (s, 1H), 5.87 (s, 1H), 2.90 (t, / = 7.5 Hz, 2H), 1.71 (m, 2H), 1.25~ 1.37 (m, 20H), 0.88 (t, J = 6.5 Hz, 3H) ppm; HRMS: m/z [ +H] ⁺ calcd for C ₂₀ H ₃₃ O ₃ ⁺ : 321.2424, found: 321.2434.

Compound I -35 : 3-(tetradecyloxycarbonyl)-1,2-phenyldiacetate

(140 mg, 0.4 mmol) of compound I-6, (49 mg, 0.4 mmol) 4-dimethylaminopyridine DMAP was dissolved in 4 ml of dry pyridine, and 1 ml of acetic anhydride was added dropwise with stirring, and stirred at room temperature for 5 h. After completion of the reaction, it was extracted with EtOAc. Silica gel column chromatography (developing solvent: n-hexane/ethyl acetate, 5/1, V/V). White solid, 130m, 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.90 (dd, / = 1.0, 7.5 Hz, 1H ), 7.37 (dd, / = 1.0, 8.0 Hz, 1H), 7.32 (t, / = 8.0 Hz, 1H), 4.26 (t, / = 6.5 Hz, 2H), 2.34 (s, 3H), 2.32 (s , 3H), 1.72 (m, 2H), 1.40 (m, 2H), 1.26~ 1.33 (m, 20H), 0.88 (t, / = 7.0 Hz, 3H); ¹³ C NMR (125 MHz, CDC1 ₃ ) δ : 168.4, 168.2, 163.9, 143.5, 142.6, 128.9, 127.6, 126.0, 125.2, 65.6, 31.9, 29.6~ 29.7, 29.5, 29.4, 28.6, 25.9, 22.7, 20.6, 14.1 ; IR (KBr) v: 2919, 2850 , 1775, 1710, 1463, 1375, 1306, 1294, 1197, 1106, 761 cm - ¹ ; HRMS: m/z [ +H] ⁺ calcd for C ₂₅ H ₃₉ 0 ₆ ⁺ : 435.2741 , found: 435.2752.

Compound I -36: 3-(Tetradecylcarbonyl)-1,2-phenyldiacetate

The synthesis method is the same as the compound 1-35, and the reaction charge is: (100 mg, 0.29 mmol) of compound 1 -23, (37 mg, 0.3 mmol) of DMAP, (0.6 ml) acetic anhydride, 2 ml of dry pyridine to obtain a pale yellow liquid 1 10 mg. 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.34~ 7.37 (m, 3H), 3.65 (t, / = 8.0 Hz, 2H), 2.26 (s, 3H), 2.24 (s, 3H), 1.52 (m, 2H), 1.21~ 1.31 (m, 22H), 0.88 (t, / = 7.0 Hz, 3H); MS (m/z): 434 [M] ⁺ .

Compound I -37 : 5-(tetradecyloxycarbonyl)benzene -1,2,3-triacetate

Synthetic method with compound I-35, the reaction charge was: (28 mg, 0.076 mmol) of compound I -17, (12 mg, 0.1 mmol) DMAP, (0.3 ml) acetic anhydride, 1 ml dry pyridine to give a white solid 29 mg . 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.80 (s, 3Η), 4.31 (t, / = 6.5 Hz, 2H), 2.32 (s, 9H), 1.75 (m, 2H), 1.21~ 1.31 (m, 22H), 0.88 (t, J = 7.0 Hz, 3H); MS (m/z): 492 [M]+.

Compound I -38 : 5-(tetradecylformyl)benzene -1,2,3-triacetate

The synthesis was carried out in the same manner as the compound I-35, and the reaction was carried out as follows: Compound I -31 (100 mg, 0.27 mmol) (37 mg, 0.3 mmol) DMAP, acetic anhydride (0.6 ml), 2 ml dry pyridine to give a white solid 1 13 mg . 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.53 (s, 2H), 6.04 (s, 1H), 3.42 (q, / = 6.5 Hz, 2H), 2.32 (s, 9H), 1.59 (m, 2H), 1.27~ 1.39 (m, 22H) , 0.89 (t, / = 7.0 Hz, 3H); MS (mz): 491 [M] ⁺ .

Compound I -39: 1,2-phenylenic acid ester

The synthesis method is the same as the compound I -4, and the reaction charge is: o-diphenol (110 mg, 1 mmol), eicosanoic acid (310 mg, 1 mmol), dicyclohexylcarbodiimide (250 mg, 1.2 mmol), 15 ml of tetrahydrofuran afforded 391 mg of a white solid. 1H NMR (500 MHz, CDC1 ₃ ) δ: 7.13~ 7.16 (t, 1H, / = 7.5 Hz), 7.09~ 7.11 (d, 1H, / = 7.5 Hz), 7.02~ 7.04 (d, 1H, J = 1.5 Hz), 6.92~ 6.95 (t, 1H, / = 7.5 Hz), 2.63 (t, 2H, / = 6.5 Hz), 2.42 (t, 2H, / = 6.5 Hz), 1.96~ 1.99 (m, 12H), 1.17~ 1.42 (m, 60H), 0.89 (bs, 6H); MS (mz): 699 [M]+.

Compound I -40: 2,3-dihydroxyphenyltetradecanoate

The synthesis method is the same as the compound I -4, and the reaction charge is: o-trisphenol (126 mg, 1 mmol), myristic acid (456 mg, 2 mmol), dicyclohexylcarbodiimide (250 mg, 1.2 mmol), 15 ml of tetrahydrofuran afforded 120 mg of a white solid. 1H NMR (500 MHz, CDCI3) δ: 6.82 (m, 1H), 6.61 (dd, J=3.0, 7.0 Hz, 1H), 6.57 (d, / = 8.0 Hz, 1H), 5.74 (s, 1H), 5.30 (s, 1H), 2.64 (m, 2H), 1.77 (m, 2H), 1.41 (m, 2H), 1.26 to 1.35 (m, 18H), 0.88 (t, / = 6.5 Hz, 3H) ppm; ¹³ C NMR (125 MHz, CDC1 ₃ ) δ: 173.2, 148.0, 146.9, 126.8, 121.1, 113.4, 112.8, 109.7, 34.4, 34.1, 31.9, 29.6~ 29.7, 29.4, 29.3, 29.2, 29.0, 24.9, 22.7, 14.1 ppm; IR (KBr) v: 3551 , 3341 , 2920, 2848, 1735, 1604, 1473, 1413, 1273, 1142 cm"¹; HRMS: mJz [M+H]+ calcd for C ₂₀ H ₃₃ O ₄ ⁺ : 337.2373, found: 337.2361.

Compound I -41 : 3- (tetradecyloxy)benzene -1,2-diol

Adding o-trisphenol (200 mg, 1.58 mmol), tetradecyl alcohol (339 mg, 1.58 mmol), triphenylphosphine (414 mg, 1.58 mmol) to anhydrous tetrahydrofuran, and adding azodicarboxylic acid dropwise in an ice bath Diethyl ester (275 mg, 1.58 mmol), reacted at room temperature overnight, the reaction was completed, the solvent was evaporated under reduced pressure, and concentrated toluene (degrading solvent: petroleum ether/ethyl acetate, 20/1, VA, pale red Solid 220 mg. 1H NMR (500 MHz, CDC1 ₃ ) δ: 6.73 (t, / = 6.5 Hz, 1H), 6.58 (dd, / = 1.0, 8.0 Hz, 1H), 6.45 (dd, / = 1.0, 8.5 Hz, 1H), 5.41 (s, 1H), 5.29 (s, 1H), 4.02 (t, / = 6.5 Hz, 2H), 1.80 (m, 2H), 1.44 (m, 2H), 1.26~ 1.36 (m , 20H), 0.88 (t, / = 6.5 Hz, 3H) ppm; ¹³ C NMR (125 MHz, CDC1 ₃ ) δ: 146.4, 144.1, 132.7, 119.7, 108.6, 104.1, 69.2, 31.9, 29.6 to 29.7, 29.4 , 29.3, 26.0, 22.7, 14.1 ppm; IR (KBr) v: 3444, 3373, 2922, 2851, 1627, 1526, 1471, 1234, 1178, 1071, 765, 717 cm- ¹ ; HRMS: mJz [M+H ]+ calcd for C ₂ .H ₃₅ 0 ₃ ⁺ : 323.2581 , found: 323.2560.

Example 2. Benzoate derivatives have the function of nerve growth factor

2-1:

 Studies have found that NGF can prevent or reduce neuronal degeneration, and promote nerve growth and neuroprotection. Since PC 12 cells have the general characteristics of nerve cells, under the action of NGF, PC12 cells will stop dividing, grow protrusions, and transform into neuron-like cells. Therefore, a compound which can cause PC12 cells to transform into neuron-like cells has an application value for preventing and treating neurodegenerative diseases such as senile dementia.

Experimental operation:

1) Culture of PC 12 cells: Connect ²⁰ ×10 ⁴ PC 12 cells in a 100 mm culture dish containing 10 ml DMEM medium (containing 10% horse serum, 5% fetal bovine serum), and replace the culture once every two days. Base, another three days to succeed. First with Wash the cells twice with PBS, add 10 ml of PBS to the culture dish, incubate in a 37 ° C, 5% C0 ₂ incubator for 10 minutes, purge, transfer to a 15 ml disposable centrifuge tube, and centrifuge the blood cells. Count on the count board. Add 24 ml of serum-containing DMEM medium to each well of the 24-well cell culture plate. After counting the cells, connect 2><10 ⁴ cells per well, and incubate in a C0 ₂ incubator for 24 hours.

2) Activity test: DMSO was used as a negative control, NGF 40 ng was used as a positive control, and Compound I was configured to a different concentration of DMSO solution. The original medium of each well of a 24-well cell plate was replaced with 1 ml of a DMEM solution containing 1% DMSO and a sample (without serum), and then placed in a 37 V, 5% C0 ₂ incubator. The morphological changes of the cells were observed every 24 hours and 6 consecutive days under an inverted microscope, and the neurite differentiation rate of the cells (the ratio of the number of cells whose neurites were longer than the diameter of the cell body to the total number of cells in the field of view) was recorded, about 100 per field of view. For each cell, 3 were randomly selected, averaged, and statistically plotted.

) Experimental results:

 Table 1. Series of compounds I after 48 hours at the optimal concentration of neurite outgrowth rate of PC 12 cells Compound optimal concentration (μΜ) Nerve protrusion

 I -1 10 37

 I -2 3 35

 I -3 3 73

 I -4 1 80

 I -5 1 82

 I -6(ABG-001) 1 87

 I -7 1 82

 I -8 3 82

 I -9 10 79

 I -10 10 70

 I -11 30 61

 I -12 3 27

 I -13 3 25

 I -14 1 36

 I -15 3 83

 I -16 0.3 80

 I -17 3 32

 I -18 3 28

 I -19 10 30

 I -20 10 33

 I -21 1 32

I -22 3 27 I -23 0.03 61

 I -24 0.03 58

 I -25 0.1 35

 I -26 0.1 50

 I -27 0.1 39

 -28 0.1 49

 -29 1 73

 -30 0.3 53

 -31 3 30

 -32 3 57

 -33 1 84

 -34 3 15

 -35 0.3 71

 -36 0.3 68

 -37 0.3 70

 -38 0.3 65

 -39 3 40

 -40 3 37

 -41 0.3 44

 Negative control (DMSO) 1% 10

 Positive control (NGF) 40ng/ml 83

 The results of preliminary pharmacological tests showed that the length of the alkyl chain, the type, position and number of substituents on the benzene ring, and the way the benzene ring was linked to the side chain had an important effect on the neurite activity of PC 12 cells. Among them, compounds 1-4, 1-5, I -6 (ABG-001), I -7, I -8, I -9, I -10, I -15, I -16 were optimally concentrated after 48 hours. The neurite differentiation rate of PC 12 cells was comparable to or higher than that of the positive control NGF, and these compounds have further research and development significance.

 Figure 1 shows the change of neurite differentiation rate of PC 12 cells with dose increase after 48 hours of addition of compound ABG-001. C: 1% DMS0 is a negative control; NGF C40ng/ml) is a positive control, compound ABG- The concentration unit of 001 is μΜ.

 Figure 2 is a photomicrograph of neurites of PC 12 cells after 48 hours of addition of compound ABG-001, Figure a, 1% DMSO as a negative control; Figure b, NGF 40 ng/ml as a positive control; Figure c, Compound ABG The concentration of -001 is 1 μΜ. 2-2:

1,25 (ΟΗ) ₂ Vitamin D3 can induce the expression of nerve growth factor (NGF) by activating its receptor. Our study found that let-hydroxylase knockout mice showed reduced expression of NGF and hippocampal neuronal regeneration compared with wild-type mice. Supplementation with 1,25(OH) ₂ vitamin D3 or NGF improved neurological regeneration in let-hydroxylase knockout mice.

Main materials of the experiment: A 12-week-old target knockout let-hydroxylase gene mouse was selected as an experimental animal model of adult NGF deficiency. (This lab was developed in cooperation with McGill University, Canada) Experimental procedure: 12-week old let-hydroxylase knockout mice were injected intraperitoneally with BU. After 10 days, brain tissue was fixed by perfusion of 4% paraformaldehyde through the left ventricle, and BrdU immunostaining was performed to label newborn neurons. ABG-001 is dissolved in

Oral administration was carried out in 99.5% ethanol, followed by dilution with physiological saline.

 Experimental results (Fig. 3): Compared with the NGF treatment group (intraventricular administration), ABG-001 intraperitoneal administration can effectively improve the nerve regeneration function of let-hydroxylase knockout mice; Figure 3 (a) It can be seen that the expression of nerve growth factor (NGF) in the hippocampus of let-hydroxylase knockout mice (-/-) is significantly lower than that of wild-type mice (+/+); as can be seen from Figure 3 (b) Compared with wild-type mice, the survival of hippocampal neonatal neurons in hydroxylase knockout mice was reduced. NGF treatment (intraventricular administration) improves the survival of neonatal neurons in hydroxylase knockout mice. Compared with the NGF-treated group, ABG-001 treatment (abdominal administration) can effectively ameliorate the neuroregeneration disorder caused by NGF deficiency.

2-3:

 On the third day after transient cerebral ischemia, cell proliferation increased in the dentate gyrus subgranules (SGZ) of the hippocampus, peaked on days 7-10, and then decreased. Recent studies have found that the vast majority of neonatal neural cells induced by cerebral ischemia die within 1-2 weeks after birth. Although cerebral ischemia activates neural stem cells and stimulates endogenous nerve regeneration, the number of precursor cell proliferation is limited, and factors that regulate neonatal neural cell migration, differentiation, survival, neuronal repair, and synapse formation are also insufficient. Endogenous nerve regeneration after cerebral ischemia does not achieve its own role in repairing neurological deficits. Therefore, how to reduce the death of newborn nerve cells, promote their migration to ischemic necrosis, induce differentiation into functional neurons, and play an endogenous repair role has become the focus of current ischemic brain injury research. The proliferation, migration and differentiation of neural stem cells are regulated by the interaction of many in vitro and in vivo environmental factors. Although the mechanism of nerve regeneration after cerebral ischemia is still unclear, neuronal regeneration after cerebral ischemia in neurotrophic factor knockout rats is significantly attenuated, and intraventricular injection of neurotrophic factors can promote stem cell proliferation and increase the number of new nerve cells.

Main materials of the experiment: Sprague-Dawley (SD) rats (10 males and females) weighing 200 g to 250 g were purchased from the Experimental Animal Center of Jiangsu Province.

 Experimental operation: 10% chloral hydrate was intraperitoneal anesthesia, the right common carotid artery and external carotid artery were ligated, the distal end of the internal carotid artery was clamped with an artery clamp, and the fishing line was inserted into the distal end of the internal carotid artery. The middle cerebral artery was applied for 60 minutes. Obstruction (middle cerebral artery occlusion in the figure referred to as MCAO), a mouse model of focal cerebral ischemia was prepared (using an electric blanket to maintain the anus temperature of the experimental animals at 37 ± 0.5 degrees). Intraperitoneal injection of ABG-001 (0.5-1.0 mg/kg) for 14 consecutive days from 48 hours after cerebral ischemia. BrdU was given on the third day after cerebral ischemia. Brain tissue was fixed by perfusion of 4% paraformaldehyde in the left ventricle 28 days after cerebral ischemia, and BrdU immunostaining was performed to label new neurons.

 EXPERIMENTAL RESULTS (Fig. 4): ABG-001 administration promotes the survival of new neurons and migration to the striatum in the brain injury area after cerebral ischemia, and is easy to restore the nervous system function after ischemic brain injury.

 Theoretical analysis and preliminary experimental results indicate that all of the other compounds in Example 1 also have similar effects as ABG-001.

Example 3 ABG-001 acute high-dose oral, long-term oral and intraperitoneal administration did not show toxicity

 The main materials of the experiment: ICR mice (body weight 22g~25g, 5 males and 5 females, purchased from Experimental Animal Center of Zhejiang University).

Experimental operation: ABG-001 is less soluble in water. (1) Dissolve ABG-001 in 99.5% ethanol, then add 1% Tween 80, diluted with physiological saline to the use concentration (the final concentration of ethanol is less than 2%). Oral medication 5g / kg. (2) intraperitoneal administration, ABG-001 was dissolved in dimethyl sulfoxide (DMSO), and then diluted with physiological saline to the use concentration (final concentration of DMSO below 1%);

 Acute poisoning test: 20 ICR male mice of 4 weeks old, male and female, were randomly divided into control group and 5g/kg treatment group. The compound ABG-001 dissolved in 1% Tween-80 was orally administered at 5 g/kg for one week. The mental state of the animals was observed every day, and the body weight and food intake were measured. After 10 minutes of compound administration, the limbs of the mice contracted and the amount of exercise decreased. After 1 hour, all returned to normal. There was no death in the mice within one week, and there was no significant change in food intake, but the change in body weight was significantly reduced. There were no significant differences in heart, liver, spleen, kidney and white adipose tissue weight and eye.

Experimental results:

 (1) Table 2 shows that compared with the saline control group, the ABG-001 treatment group did not show weight gain/decrease (including the weight of each organ), nor did breathing (50 to 60 times/min) and heart rate ( 305 to 400 beats/min), abnormality of mean arterial pressure (210 mmHg), and symptoms such as lethargy, mania, and abnormal motor behavior.

 Table 2: Blood biochemical parameters of ICR mice (early =10, =10 in each group; ABG-001=5g/kg)

 |

 GLO

TP ALB ^

 B ALT AST TBIL DBIL CHE

 Group (g/L (g/L A/G)

 (g/L (U/L) (U/L) (umol/L) (umol/L) (U/L) ) )

 )

 56.86 40.20 16.66 2.42 31.00

1.62 0.82 9350.80 士 士 士 士 士 Early

1.62 0.82 9350.80 SST

 Show ±0.34 ± 0.09 ± 156.32

0.43 0.37 0.32 0.06 2.28

 57.54 39.00 18.54 2.12 33.40

 As early as 111.40 1.62 0.96 10075.40, a gentleman

 5g/kg ± 15.80 ± 0.16 ± 0.08 ± 411.98

 1.24 1.30 0.98 0.14 1.44

 57.40 36.50 20.90 1.75 37.75

 Inch 88.75 1.48 0.87 6081.50 Judges

 Show ± 12.16 ±0.33 ± 0.11 ± 184.27

2.11 0.86 1.33 0.09 2.49

 54.68 18.68 1.94 32.00

 75.40 1.64 0.91 5974.60 士士士士士

 5g/kg ±2.34 ± 0.27 ± 0.04 ± 153.90

 0.88 0.24 0.07 1.10 In Table 2, TP (total protein), ALB (albumin), GLOB (globulin), ALT (alanine aminotransferase), AST (aspartate aminotransferase), TBIL (total bilirubin), DBIL (direct bilirubin), CHE (acetylcholinesterase) (2) Compared with the saline control group, the ABG-001 5g/kg group did not show abnormalities in blood biochemical indicators (Table 3), suggesting liver, kidney, Functions such as hematopoietic organs are not impaired.

Table 3: Body weight and organ coefficients of ICR mice (10 per group, =5, early = 5; ABG-001 = 5 g/kg) Group weight (g) Heart (%) Liver (%) Kidney (%) Spleen (%) Fat (%)

24.64 0.685 2.102 0.684 3.185

3⁄4 control group

 ±0.38 ±0.086 ±0.148 ±0.078 ±0.260

23.67 0.607 7.904 2.200 0.698 2.129

5g/kg group ±0.86 ±0.044 ±0.289 ±0.110 ±0.036 ±0.105

20.30 0.658 6.669 1.681 0.607 2.848 Early control group

 ±0.56 ±0.031 ±0.205 ±0.046 ±0.038 ±0.384 Early 19.12 0.726 6.373 1.756 0.701 2.095

5g/kg group ±1.41 ±0.037 ±0.234 ±0.047 ±0.039 ±0.405 Organ coefficient=organ weight/weight (%)

 Chronic toxicity test: ABG-001 was administered by intraperitoneal injection (10 mg/kg/day) for 60 consecutive days or by oral administration (10 mg/kg/day) (administered as saline as a control group). Abdominal anesthesia with 10% chloral hydrate was used to record respiration, heart rate, and mean arterial pressure. Blood is then taken from the left ventricle for testing of hematology and blood biochemical indicators. And take the internal organs to take observations of histology.

Experimental results:

 (1) Table 4 shows that there was no weight gain/decrease (including the weight of each organ) in the ABG-001 treated group compared with the saline control group. There were also no abnormalities in breathing (50 to 60 beats/min), heart rate (305 to 400 beats/min), average arterial pressure (210 mmHg), and lethargy, arrogance, and abnormal motor behavior.

Table 4: Mouse body weight and various organ coefficients (Ϋ=10, ί=10 in each group; ABG-001=10 mg/kg gavage) Group weight (g) Lung (%) Heart (%) Liver (%) Kidney (%) spleen (%)

36.50 0.71 0.502 5.093 1.302 0.419 Control group

 ±2.84 ±0.21 ±0.10 ±0.32 ±0.20 ±0.12 Solvent 33.20 0.82 0.605 5.128 1.235 0.405

 (gavage) ±2.72 ±0.24 ±0.19 ±0.43 ±0.17 ±0.09 Solvent 36.22 0.78 0.524 4.912 1.193 0.368

(abdominal cavity) ±2.47 ±0.12 ±0.16 ±0.55 ±0.18 ±0.14 ABG-001

 35.83 0.87 0.482 5.243 1.203 0.390 (gavage)

 ±2.11 ±0.28 ±0.22 ±0.67 ±0.32 ±0.07 lOmg/kg

 ABG-001

 32.30 0.80 0.523 5.031 1.133 0.371 (abdominal cavity)

 ±3.42 ±0.19 ±0.15 ±0.37 ±0.15 ±0.09 5mg/kg organ coefficient = organ weight / weight

 (2) Compared with the saline control group, the chronic ABG-001 administration group showed no abnormality in the blood biochemical index (Table 5), suggesting that the liver, kidney, and hematopoietic organs were not damaged.

Table 5: Main blood biochemical indicators of mice (Ϋ10 for each group = 10; ABG-001= ^: 10 mg/kg for gavage)

 GLO

TP ALB ALT TBIL DBIL

B AST CHE

 Group (g/ (g/L A/G (U/L (umol (umol/L)

 (g/L (U/L) (U/L)

L) ) ) /L) )

 )

 57.2 38.5 17.7 32.2 85.4 7863.2

 2.14 1.71

 8 2 9 5 2 0.71 4 Control group ±0.0 ± 0.2

 ±0.6 ±0.5 ±0.9 ±1.4 ±15. ±0.10 ±120.6

 8 4

 9 8 2 3 2 7

56.2 39.9 19.1 32.8 111. 8017.4

 2.01 1.58

 Solvent 3 4 4 5 41 0.92 0

 ±0.0 ±0.1

 (turn) ±1.0 ±0.9 ±1.2 ±1.4 ±17. ±0.08 ±168.5

 7 9

 4 7 1 4 10 4

57.1 36.5 18.4 33.6 75.7 7684.3

 1.91 1.63

 Solvent 0 7 3 1 8 0.85 8

 ±0.1 ±0.2

 (abdominal cavity) ±1.1 ±0.6 ±0.8 ±1.9 ±14. ±0.09 ±176.5

 3 6

 1 7 1 9 72 9

ABG-00 55.7 38.2 20.0 33.0 85.6 8344.7

 2.04 1.70

 1 (gavage) 8 5 2 4 3 0.91 5

 ±0.0 ±0.1

 10mg/k ±0.6 ±1.0 ±1.0 ±1.0 ±11. ±0.05 ±191.8

 8 9

 g 0 2 4 3 41 3

 56.8 37.4 18.3 32.1 89.4 7976.4

ABG-00 1.88 1.51

 9 3 5 9 7 0.89 3

1 (abdominal cavity) ±0.0 ±0.3

 ±1.1 ±0.7 ±0.8 ±1.5 ±15. ±0.09 ±143.0

5mg/kg 9 5

7 2 7 4 98 6 Table 5, TP (total protein), ALB (albumin), GLOB (globulin), ALT (alanine aminotransfer) Enzyme), AST (aspartate aminotransferase), TBIL (total bilirubin), DBIL (direct bilirubin), CHE (acetylcholinesterase). Theoretical analysis and preliminary experimental results indicate that all of the other compounds in Example 1 also have similar effects to ABG-001. Example 4 ABG-001 can pass the blood-brain barrier (Figure 5)

 Main materials of the experiment: Sprague-Dawley (SD) rats (10 males and females) weighing 200 g to 250 g were purchased from Jiangsu Experimental Animal Center.

 Experimental operation: 10% chloral hydrate was intraperitoneally anesthetized. The microelectrode was inserted into the pyramidal cell layer of hippocampal CA1 region according to the coordinates of 0.3 mm, 1.0 mm, and 2.5 mm depth, and the spontaneous discharge frequency of hippocampal CA1 neurons was recorded. Baseline stable recording After 20 minutes, ABG-001 was administered intraperitoneally and recorded continuously for 1-2 hours. Analysis of whether ABG-001 regulates the excitability of hippocampal neurons through the blood-brain barrier.

 Experimental results (Fig. 5): After 20-40 minutes of intraperitoneal injection of ABG-001 (0.5-lmg/kg), the spontaneous discharge frequency of hippocampal CA1 neurons increased significantly and lasted for more than 2 hours, suggesting that ABG-001 can pass blood. The brain barrier regulates the excitability of nerve cells.

 Theoretical analysis and preliminary experimental results indicate that all of the other compounds in Example 1 also have similar effects as ABG-001. Example 5, ABG-001 can promote the regeneration of brain neurons through the blood-brain barrier (Fig. 6)

 In 1988, Erikssion et al. used BrdU (5-bromodeoxyuridine) to label mitotic cells and found that neural stem cells of adult mammalian hippocampus can differentiate into neuronal-neurogenesis. Neonatal neurons in the dentate gyrus of adult hippocampus are similar to mature granulosa cells and can establish synaptic connections with synaptic plasticity in CA3 neurons. Adult neurogenesis can replace neuronal death due to natural aging or disease, maximizing brain function and structural integrity. Studies have shown that the reduction of nascent cytokines in the aged brain (neural neonatal decline) is associated with senile cognitive decline. This study focused on the effects of ABG-001 on neurogenesis (mainly including neural stem cell proliferation, survival, differentiation).

 Main materials of the experiment: 2 months old Kunming mice (40 males and females) weighing 25g~30g were purchased from Jiangsu Experimental Animal Center.

Experimental procedure: 1 Cells with mitosis were labeled with BrdU (5-bromodeoxyuridine), and ABG-001 (0.5 mg/kg/d) was administered for 14 consecutive days after BU injection. Then, BrdU-positive (BrdU ⁺ ) cells in the hippocampal DG region were detected by immunohistochemistry at 48 hours, sputum days and 28 days after the first dose of BrdU to determine the proliferation of ABG-001 (24-hour-BrdU ⁺ Effects of cells (7 days old - BrdU ⁺ cells) and maturation (28 days old - BrdU ⁺ cells). 2 Immunostaining with neuron-specific nuclear protein (NeuN) and glial cells (GFAP) was used to observe the effect of ABG-001 on neonatal differentiation. 3 Immunostaining with Doublecortin (DCX) to observe the effect of ABG-001 on the growth of neonatal neurons. Administration of ABG-001: ABG-001 was dissolved in 99.5% ethanol or dimethyl sulfoxide, and then diluted intraperitoneally with tea oil or diluted with physiological saline for oral administration.

Experimental results (Fig. 6): Oral or intraperitoneal administration of ABG-001 can promote the proliferation of hippocampal dentate gyrus neural stem cells (a), promote the growth of new neurons (b), and promote the differentiation of stem cells into neurons (c). Tip ABG-001 has God Through nutrition.

 Theoretical analysis and preliminary experimental results indicate that all of the other compounds in Example 1 also have similar effects as ABG-001. Example 6. ABG-001 has anti-aging effect (Figure 7)

6-1:

 Mammals obtain galactose from lactose, which is hydrolyzed in the body to produce glucose and galactose, while galactose is rapidly digested into glucose in the liver. Excess galactose is reduced to galactitol by aldose reductase catalysis. Galactose is a metabolic end product that cannot be further catabolized and accumulates, increasing the osmotic pressure of cells, leading to cell swelling, dysfunction and metabolic disorders, which eventually lead to aging of the body. A large number of studies have reported D-gal galactose-induced aging animal models. D-gal causes rat embryonic brain neurons to develop degenerative changes such as slow growth, neurite outgrowth, and increased mortality. D-gal leads to a decrease in the number of in vitro division algebras in rat lung fibroblasts, a decrease in G2-M phase cells, an increase in G-G1 phase cells, and a slower cell proliferation rate in the diploid fibroblast dividing cycle. D-gal inhibits cell growth and development and reduces the number of divisions, accelerating cell senescence.

 Main materials of the experiment: Kunming mice of 2 months old (30 males and 30 females) weighing 25g~30g were purchased from the Experimental Animal Center of Jiangsu Province.

Experimental procedure: D-gal galactose (100 mg/kg/day) was administered intraperitoneally for 60 days. After 6 weeks, the mice were slow-moving, with dark coats and thin bodies, showing obvious signs of aging, proving that the aging model was successfully established. After 20 days of D-gal galactose injection, ABG-001 was orally administered (0.1, 5.0, 10.0 mg/kg/day) or intraperitoneally (0.1, 0.5, 5 mg/kg/day). When D-gal was injected for 30 days, BrdU (5-bromodeoxyuridine, 50.0 mg/kg) was administered 4 times at intervals of 6 hours. Then, 28 days after the administration of BrdU, brain tissue was fixed by perfusion of the left ventricle with 4% paraformaldehyde, and BrdU immunostaining was performed to label the newborn neurons. BrdU-positive (BrdU ⁺ ) cells in the hippocampal DG region were examined to determine the effect of ABG-001 on neonatal cell survival and maturation (28-day-brown U ⁺ cells). The effect of ABG-001 on the growth of neonatal neurons was observed by immunostaining with Doublecortin (DCX). The mice were sacrificed on the 2nd day after the last administration, and the hippocampus and cerebral cortex were taken out to measure glutathione peroxidase (GSH2Px), superoxide dismutase (SOD), and monoamine oxidase (monoamine oxidase). MAO) Biochemical indicators of aging such as total anti-oxidation competence (T-ACO).

Experimental results (Fig. 7): (1) Long-term injection of D-gal galactose can significantly reduce the number of new neurons and impair the growth of new neurons (a); (2) D-gal galactose-aging mice There was no obvious abnormality in motor function (b), but spatial learning and memory function were significantly reduced; (3) ABG-001 treatment improved D-gal-induced survival and neurite outgrowth of neonatal neurons (a); (4) ABG- 001 treatment can improve the memory function of mice with brain aging (c) and (d); (5) biochemical indicators of oxidative stress in mice (early group = 10, 3 = 10; ABG-001 = 10 mg / kg) ) shows that the improvement in total antioxidant capacity is not obvious (e), (f), (g) and (h). The main materials of the experiment: SAM R1 (6-week-old males, 18 each) and SAM P8 (9-week-old males, 18 each) were purchased from the First Affiliated Hospital of Tianjin Medical University. SAM P8 mice are a rapidly aging animal model. Experimental procedure: SAM P8 9-month-old mice were randomly divided into 3 groups, control group (1% Tween 80 saline), low dose (lmg/kg) group and high dose (3 mg/kg) group, each group 6 only. In addition, SAM R1 six-week-old mice served as a youth control group for normal aged mice. Each group was intraperitoneally injected with 1% Tween 80 saline or ABG-001 daily for two weeks. Detection of learning and spatial memory functions. (1) Y-maze experiment: Spontaneous alternation behavior can be assessed using the Y-maze experiment. Y-maze is a black Y-shaped three-part radiant lost box, consisting of 3 arms and a connecting zone. The three arms are at an angle of 120° to each other, each arm is 38.5 cm long, 8 cm wide and 3.5 cm wide. , 12 cm high, one arm can be selected as the starting arm. Each mouse was released from the end of the fixed arm, and the mice were allowed to move freely for 8 minutes, and the order of each arm was recorded. Three consecutive entries into the three different arms were defined as alternating arms (eg ABC, ACB, BAC, BCA, CAB, CBA). The calculation of the alternate arm rate is: 1 (number of errors / total number of times 2) χ 100%. In addition, the total number of arms can be determined. (2) Novel object discrimination experiment: The novel object discrimination experiment is based on the ability of rodents to distinguish familiar and novel objects. The device used was a white uncovered box (length X width X height: 40 x 20 x 18 cm). First, there is no object in the box. Each time the mouse is released from the same position, let it move freely for 5 minutes to adapt to the environment, called the adaptation period, for two days. The third day is the familiarization period and the test period. Familiarization period: After placing two identical objects, put the mouse in, freely move for 5 minutes, and count the mouse nose as less than 2 cm from the object or on the recognized object, and record the two pairs. The exploration time of the same object. Test period: After 1 hour interval, replace with two different objects (one of which is the same object as the familiar period (0, the other is a novel object (n)), let it move freely for 5 minutes, and record the mouse pair Familiar with the exploration time of the object f and the exploration time of the novel object n. The familiar period and the test period are combined by two kinds of objects and positions according to random numbers. The calculation formula of the cognitive index is the time n/total exploration of the novel object. Time n+f.

 Experimental results (Fig. 8): (1) Compared with SAM R1 mice, there was no significant change in the total number of arms and alternate arm in the SAM P8 mice (a); (2) receiving high or low doses of ABG- 001-treated SAM P8 mice showed significantly enhanced learning and memory function compared to untreated SAM R1 mice and SAM P8 mice (b); (3) SAM P8 mice versus novelty objects compared to SAM R1 mice The discriminating ability was significantly reduced (c); (4) ABG-001 treatment was able to restore the discrimination ability of SAM P8 mice to novel objects in a concentration-dependent manner (d).

 Theoretical analysis and preliminary experimental results indicate that all of the other compounds in Example 1 also have similar effects as ABG-001. Example 7. ABG-001 has the effect of preventing and treating senile dementia (Fig. 9)

74

Main materials of the experiment: Sprague-Dawley (SD) rats (20 males and females) weighing 200 g to 250 g were purchased from Jiangsu Experimental Animal Center. 1 Using stereo positioning technology, the osmotic pump (osmotic minipump: Alzet 2002; Alze, CA) is connected to the cannula and fixed in the lateral ventricle of one side (0.3mm behind the anterior iliac crest, 1mm on the right side, deep) 2.5mm). Αβ _{( 1} _42) was perfused into the lateral ventricle for 2 weeks (100 pM/day). The stability of the AD model was verified by observing the deposition of Αβ, spatial memory function, and cholinergic system function.

Experimental procedure: ABG-001 (0.5-1.0 mg/kg) was administered intraperitoneally for 14 consecutive days starting on the 2nd day after Αβ ₍₁ _42) perfusion. A water maze test was performed on the 7th day after Αβ α-4 _{2 )} perfusion to examine the spatial memory function. The trajectory of the rat was recorded and the staging latency was calculated. 15 days after Αβ _α _ ₄₂₎ perfusion, brain tissue was fixed by perfusion of 4% paraformaldehyde through the left ventricle. After the paraffin is embedded, the hippocampus is serially sliced. After staining with 1% toluidine blue, the density of viable pyramidal cells in the CA1 region of the hippocampus was measured (the number of granule cells per mm length of the pyramidal cell layer).

Experimental results Figure 9: Αβ _α _ ₄₂₎ perfusion caused about 60% loss of neuronal death in the hippocampal CA1 region, leading to prolonged escape latency, suggesting a reduction in spatial cognitive function. ABG-001 treatment can significantly reduce the death of hippocampal neurons in Αβ _α _ ₄₂₎ dementia rats, and improve the spatial learning and memory function of Αβ rats.

ABG-001 treatment improves endogenous neurological regenerative disorders in Alzheimer's disease (AD)

 The neural stem cells of the adult mammalian hippocampal dentate gyrus can differentiate into neural cells called neurogenesis. Adult nerves are thought to replace and repair neuronal loss due to natural aging or disease, maximizing the structure and function of the brain. Alzheimer's disease (AD) is a neurodegenerative disorder characterized by progressive cognitive dysfunction. Whether newborn neurons can replace diseased neurons and improve cognitive dysfunction of AD has become a new target for AD research. The study found that the differentiation ratio of precursor cells to neurons in AD brain hippocampus was significantly reduced, while the survival rate of newborn neurons was significantly reduced, and the neurite outgrowth of newborn neurons was abnormal. These studies have confirmed that the nerve regeneration process of AD brain is seriously damaged, suggesting that nerve regeneration disorder may be one of the important pathological mechanisms of progressive cognitive decline in AD.

Main materials of the experiment: APP/PS1 transgenic AD mice (10-month-old male and female mice) (purchased at the Animal Model Center of Nanjing University). Experimental procedure: Cells with mitosis were labeled with BrdU (5-bromodeoxyuridine) and administered with ABG-001 (0.5 mg/kg/day) for 14 consecutive days after BU injection. Then, BrdU-positive (BrdU ⁺ ) cells in the hippocampal DG region were detected by immunohistochemistry 28 days after the first administration of BrdU. The effects of ABG-001 on neonatal differentiation were observed by immunostaining of neuron-specific nuclear protein (NeuN) and glial cells (GFAP). Doublecortin (DCX) is specifically expressed during the migration of neural precursor cells during the development of the central nervous system. DCX, a marker of neuronal precursor cells, can be used to study the proliferation and migration of neuronal precursor cells. The growth of neonatal neuronal processes can be observed by DCX staining. Mice were sacrificed on the 2nd day after the last administration, and hippocampus was taken out to measure acetylcholinesterase (AChE) activity.

 Experimental results (Fig. 10): 28-day-brdU+ cells of the dentate gyrus of APP/PS1 mice showed a 50% reduction compared to wild-type mice (a); the number and length of neonatal neuronal processes were significantly reduced (b). ABG-001 treatment improved the 28-day-old B U+ cells in APP/PS1 mice (a) and protected neurite outgrowth (b), suggesting that ABG-001 has improved brain neuronal regeneration and cognitive dysfunction in AD. effect. There was a slight increase in acetylcholinesterase (AChE) activity in hippocampus of APP/PS mice, and ABG-001 treatment significantly reduced hippocampal acetylcholinesterase (AChE) activity in AD mice (c).

 Theoretical analysis and preliminary experimental results indicate that all of the other compounds in Example 1 also have similar effects as ABG-001. Example 8, preparation method of ABG-001 tablet

 Prescription:

 ABG-001 100g

Starch 40g Microcrystalline cellulose 80g

 Magnesium stearate 3.0g

 Hydroxypropyl methylcellulose (E-30) (40% solution)

 Made into 1000 pieces

Method: Prepare 4% propylmethylcellulose (E-30) solution and set aside. 10 g of starch was weighed and dried at 105 ° C for 5 hours. 20 g of starch and a prescribed amount of the invented ABG-001, microcrystalline cellulose were weighed, mixed, and pulverized through an 80 mesh sieve. The material was made into a soft material using 4% propylmethylcellulose (E-30) solution, granulated with a 20 mesh sieve, and dried at 50 ° C to 60 ° C until the moisture in the granules was about 3%. After 20 mesh sieves, add the prescribed amount of dry starch (dried at 105 °C for 5 hours), magnesium stearate, and finally mix, measure the intermediate content, and fix the tablet weight;

Example 9. Preparation method of ABG-001 injection

Prescription:

 ABG-001 10g

 Propylene glycol 500mL

 Water for injection 500mL

 Made lOOOOmL

Method: Weigh the prescribed amount of ABG-001 and propylene glycol, add 500 mL of water for injection, stir to dissolve; add 0.1% activated carbon to the above solution, stir, leave for 15 minutes, decarburize the 5 micron titanium rod, and then pass the cartridge filter Fine filtration of 0. 45 micron and 0.22 micron microfiltration membrane; potting in a 10 ml ampoule and steam sterilization at 100 °C for 45 minutes, the injection of the invented ABG-001.

Claims

Claim A use of a benzoic acid ester and a derivative thereof for the preparation of a medicament for preventing and treating a neurodegenerative disease, characterized in that the benzoic acid has the following structural formula:

Wherein R is a linear, branched, saturated and unsaturated fluorenyl group having from 1 to 30 carbon atoms, an aliphatic ring or a derivative thereof; each independently selected from the group consisting of hydrogen, a hydroxyl group, a carboxyl group, an aldehyde group, an ester group, Fluorine, chlorine, bromine, iodine, sulfhydryl, amino, amide, cyano, nitro, sulfonate, trifluoromethyl, propenyl, fluorenyl, decyloxy, substituted benzyl, substituted phenyl, aryl , a heteroaryl group, a sugar group or an amino acid residue group; X is selected from C (0) 0, OC ( 0), C (0) NH, NHC (0), C (0), CH 2, S or 0.  2. Use according to claim 1, characterized in that said R is a linear, branched, saturated and unsaturated sulfhydryl group having from 6 to 22 carbon atoms, an aliphatic ring or a derivative thereof. 3. Use according to claim 1 wherein the benzoate and its derivatives are: 2,3-dihydroxybenzoic acid ethyl ester, 2,3-dihydroxybenzoic acid amyl ester, 2 , octyl 3-dihydroxybenzoate, decyl 2, 3-dihydroxybenzoate, dodecyl 2, 3-dihydroxybenzoate, tetradecyl 2, 3-dihydroxybenzoate, 2, 3 - hexadecanoyl dihydroxybenzoate, octadecyl 2, 3-dihydroxybenzoate, decyl 2, 3-dihydroxybenzoate, behenyl 2, 3-dihydroxybenzoate, Tridecyl 2,3-dihydroxybenzoate, tetradecyl 2,6-dihydroxybenzoate, behenyl 2,6-dihydroxybenzoate, tetradecyl 2,4-dihydroxybenzoate Ester, tetradecyl 3, 4-dihydroxybenzoate, tetradecyl 3,4,5-trihydroxybenzoate, tetradecyl 2-hydroxybenzoate, tetradecyl 3-hydroxybenzoate, Tetradecyl 2, 3-dimethoxybenzoate, octadecyl 2,3-dimethoxybenzoate, tetradecyl 2-hydroxy-3-methoxybenzoate, 2-chlorobenzene Tetradecyl formate, 2, 3-dihydroxy-N-tetradecyl Benzoylamide, 2,3-dihydroxy-N-octylbenzamide, 2,3-dihydroxy-N-dodecylbenzamide, 2,3-dihydroxy-N-hexadecanylbenzene Formamide, 2, 3-dihydroxy-N-octadecylbenzamide, 2,3-dimethoxy-N-tetradecylbenzamide, 3,4-dihydroxy^-tetradecane Benzobenzamide, 3, 4, 5-trihydroxy-N-tetradecylbenzamide, N-(3,4-dimethoxyphenyl)tetradecylideneamide, N- (3, 4-dihydroxyphenyl)tetradecylidene, 1-(3,4-dimethoxyphenyl)tetradecano-1-one, 1-(3,4-dihydroxyphenyl)tetradecene Indole-1-one, 3-(tetradecyloxycarbonyl)-1,2-phenyldiacetate, 3-(tetradecanoyl)-1,2-phenyldiacetate, 5-(tetradecyloxycarbonyl)benzene-1, 2,3-triacetate, 5-(tetradecylamyl)benzene-1, 2,3-triacetate, 1, 2- Phenyl phenyl Acid ester, 2, 3-dihydroxyphenyltetradecanoate, 3-(tetradecyloxy)benzene-1,2-diol.  A use of a benzoic acid ester and a derivative thereof for the preparation of a medicament for preventing and treating anti-brain aging, characterized in that the benzoic acid ester and its derivative have the following structural formula:

Wherein R is a linear, branched, saturated and unsaturated fluorenyl group having from 1 to 30 carbon atoms, an aliphatic ring or a derivative thereof; each independently selected from the group consisting of hydrogen, a hydroxyl group, a carboxyl group, an aldehyde group, an ester group, Fluorine, chlorine, bromine, iodine, sulfhydryl, amino, amide, cyano, nitro, sulfonate, trifluoromethyl, propenyl, fluorenyl, decyloxy, substituted benzyl, substituted phenyl, aryl , a heteroaryl group, a sugar group or an amino acid residue group; X is selected from C (0) 0, OC ( 0), C (0) NH, NHC (0), C (0), CH 2, S or 0.  5. Use according to claim 4, characterized in that said R is a linear, branched, saturated and unsaturated sulfhydryl group having from 6 to 22 carbon atoms, an aliphatic ring or a derivative thereof.  The use according to claim 4, wherein the benzoate and its derivative are: 2,3-dihydroxybenzoic acid ethyl ester, 2,3-dihydroxybenzoic acid amyl ester, 2 , octyl 3-dihydroxybenzoate, decyl 2, 3-dihydroxybenzoate, dodecyl 2, 3-dihydroxybenzoate, tetradecyl 2, 3-dihydroxybenzoate, 2, 3 - hexadecanoyl dihydroxybenzoate, octadecyl 2, 3-dihydroxybenzoate, decyl 2, 3-dihydroxybenzoate, behenyl 2, 3-dihydroxybenzoate, Tridecyl 2,3-dihydroxybenzoate, tetradecyl 2,6-dihydroxybenzoate, behenyl 2,6-dihydroxybenzoate, tetradecyl 2,4-dihydroxybenzoate Ester, tetradecyl 3, 4-dihydroxybenzoate, tetradecyl 3,4,5-trihydroxybenzoate, tetradecyl 2-hydroxybenzoate, tetradecyl 3-hydroxybenzoate, Tetradecyl 2, 3-dimethoxybenzoate, octadecyl 2,3-dimethoxybenzoate, tetradecyl 2-hydroxy-3-methoxybenzoate, 2-chlorobenzene Tetradecyl formate, 2, 3-dihydroxy-N-ten Mercaptobenzamide, 2,3-dihydroxy-N-octylbenzamide, 2,3-dihydroxy-N-dodecylbenzamide, 2,3-dihydroxy-N-hexadecene Benzobenzamide, 2, 3-dihydroxy-N-octadecylbenzamide, 2,3-dimethoxy-N-tetradecylbenzamide, 3,4-dihydroxy^-ten Tetradecylbenzamide, 3,4,5-trihydroxy-N-tetradecylbenzamide, N-(3,4-dimethoxyphenyl)tetradecylideneamide, N- ( 3, 4-dihydroxyphenyl)tetradecanoylamide, 1-(3,4-dimethoxyphenyl)tetradecano-1-one, 1-(3,4-dihydroxyphenyl) Tetradecyan-1-one, 3-(tetradecyloxycarbonyl)-1,2-phenyldiacetate, 3-(tetradecanoylformyl)-1,2-phenyldiacetate Salt, 5-(tetradecyloxycarbonyl)benzene-1, 2,3-triacetate, 5-(tetradecylamyl)benzene-1, 2,3-triacetate, 1, 2-phenylphenylecosate, 2,3-dihydroxyphenyltetradecanoate, 3-(tetradecyloxy)benzene-1,2-diol.