CN110776454A - Synthesis method of bufotenine and quaternary ammonium salt thereof and application of bufotenine and quaternary ammonium salt thereof in preparation of analgesic and anti-inflammatory drugs - Google Patents
Synthesis method of bufotenine and quaternary ammonium salt thereof and application of bufotenine and quaternary ammonium salt thereof in preparation of analgesic and anti-inflammatory drugs Download PDFInfo
- Publication number
- CN110776454A CN110776454A CN201911259232.8A CN201911259232A CN110776454A CN 110776454 A CN110776454 A CN 110776454A CN 201911259232 A CN201911259232 A CN 201911259232A CN 110776454 A CN110776454 A CN 110776454A
- Authority
- CN
- China
- Prior art keywords
- bufotenine
- quaternary ammonium
- ammonium salt
- methanol
- tryptamine
- 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.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/04—Indoles; Hydrogenated indoles
- C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
- C07D209/14—Radicals substituted by nitrogen atoms, not forming part of a nitro radical
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pain & Pain Management (AREA)
- Pharmacology & Pharmacy (AREA)
- Rheumatology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Indole Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a method for synthesizing bufotoxin tryptamine and quaternary ammonium salt thereof, which has the advantages of simple steps, safe and convenient operation, high reaction efficiency, easy purification and separation, mild conditions, low cost, total reaction yields of the bufotoxin tryptamine and the quaternary ammonium salt thereof of more than 65 percent and 60 percent respectively, high purity (>99.0 percent) of synthesized products and suitability for industrial production. Through a large number of experimental screens, combined with the behavioral evaluation of thermal pain and mechanical pain and the phospholipidomics evaluation, the activity comparison of several tryptamine components shows that the bufotoxin tryptamine quaternary ammonium salt has the optimal analgesic effect and has the synergistic effect with morphine.
Description
Technical Field
The invention relates to the field of chemical synthesis, in particular to a synthetic method of bufotenine and quaternary ammonium salt thereof, and application of the bufotenine and the quaternary ammonium salt thereof in preparation of analgesic and anti-inflammatory drugs.
Background
The N, N-methylated product of serotonin (bufotenine) has a structure shown in formula (1), and the quaternary ammonium salt thereof has a structure shown in formula (2).
Bufotamine, which is an N, N-dimethylation product of 5-hydroxytryptamine, is known as bufotamine, and is found in frogs and toad skins, mushrooms, higher plants and mammals, since it is originally found in toads; there are also reports of schizophrenia in the brain, plasma and urine. Toad venom contains toad tryptamine and its quaternary ammonium salt, and the original acid radical form of toad tryptamine quaternary ammonium salt in traditional Chinese medicine toad venom is not clear because strong acid (such as trifluoroacetic acid) and alkaline solvent are often used in the conventional chromatographic separation and purification research, so that acid radical substitution is already generated. Considering that the existence forms of different acid radicals and different basic groups of the animal salt type medicine can influence the in-vivo bioavailability, the efficacy strength and the safety of the medicine. The activity of hydrochloride of the bufotamine quaternary ammonium salt is not reported in the literature. The invention takes a mouse as a model, combines the behavioral evaluation of thermal pain and mechanical pain with the phosphatidics to carry out the research on the anti-inflammatory and analgesic effects of the bufotamine and the hydrochloride of the bufotamine quaternary ammonium salt, and provides experimental basis for the analgesic and anti-inflammatory effects of the components.
The method for chemically synthesizing the toad venom tryptamine substances is especially important for safely, efficiently and environmentally synthesizing the toad venom tryptamine and the quaternary ammonium salt thereof by researching the synthesis conditions of the toad venom tryptamine substances. Bufotamine was synthesized as early as 2007 by Chinpiao Chen's project group, which designed a seven-step Synthesis route (Wang, Y. -Y., & Chen, C. (2007). Synthesis of a Deurierium-Labelled standard of bufotenine (5-HO-DMT). Journal of laboratory Compounds and Radiopharmaceuticals,50(14), 1262. the third generation of bufotamine was carried out by the same group as the first generation of bufotamine. The route takes 3-methyl-4-nitrobenzene as a raw material, an important intermediate 5-benzyloxy indole is obtained through a series of reactions, and the bufotoxin tryptamine is obtained through the reactions such as substitution, reduction, deprotection and the like. The whole route realizes the synthesis of the bufotenine with the theoretical total yield of 23.9 percent, but part of reagents are extremely toxic (hydrazine hydrate) and do not accord with the green chemical concept, so the synthesis route is not suitable for industrial production.
Disclosure of Invention
The invention aims to solve the defects of the prior art and provides a high-efficiency synthesis method of bufotenine and quaternary ammonium salt thereof. The invention uses 5-benzyloxy indole as initial material to synthesize bufotoxin tryptamine and its quaternary ammonium salt. The preparation method has the advantages of few and simple steps, safe and convenient operation, mild conditions, low cost, high reaction efficiency, easy purification and high product purity, and is suitable for industrial production.
The invention provides a preparation method of bufotoxin tryptamine, which comprises the following steps:
a. in an organic solvent, 5-benzyloxy indole (S-0) is used as a starting material, and 2- (5- (benzyloxy) -1H-indol-3-yl) -2-oxo-acetyl halide (S-1) is generated under the acylation action of oxalyl halide, wherein X is halogen;
b. mixing the 2- (5- (benzyloxy) -1H-indol-3-yl) -2-oxoacetoacetyl halide produced above with dimethylamine or a salt thereof in a solvent in the presence of a base to react and obtain 2- (5- (benzyloxy) -1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide (S-2);
c. catalytically hydrogenating the 2- (5- (benzyloxy) -1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide produced above in an organic solvent in the presence of a hydrogenation catalyst to remove benzyl groups to give 2- (5-hydroxy-1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide (S-3);
d. the generated 2- (5-hydroxy-1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide reduces ortho carbonyl to methylene under the action of a reducing agent to obtain 3- (2- (dimethylamino) ethyl) -1H-indol-5-ol (S-4, namely bufotenine).
The invention also provides a preparation method of the bufotamine quaternary ammonium salt, which is used for preparing the bufotamine through the method and further performing the step e:
e. reacting the generated 3- (2- (dimethylamino) ethyl) -1H-indol-5-ol with a methylating agent in an organic solvent to obtain the 2- (5-hydroxy-1H-indol-3-yl) -N, N, N-trimethylethan-1-amine chloride.
In the step a, X in S-1 is preferably chlorine or bromine; the oxalyl halide is selected from oxalyl chloride and oxalyl bromide, preferably oxalyl chloride, wherein the molar ratio of S-0 to oxalyl halide is 1: 1-1: 5, preferably 1: 1-1: 3. The organic solvent is anhydrous organic solvent selected from C
1-6Alcohol, C
1-6Ethers or cyclic ethers, C
1-6Esters, C
1-6Ketones, C
1-6Halogenated alkanes, C
1-6One or more of alkyl nitrile and DMF, preferably one or more of ethyl acetate, dichloromethane, methanol, diethyl ether, tetrahydrofuran, DMF, ethanol, acetonitrile and acetone solution, more preferably ethyl acetate, diethyl ether and tetrahydrofuran. The reaction time is 0.25-3 h, preferably 0.5-1 h. The temperature is 0-80 deg.C, preferably 10-50 deg.C, more preferably 20-40 deg.C, and most preferably room temperature.
In the above step b, the base is selected from the group consisting of alkali metal hydroxides, alkaline earth metal hydroxides, carbonates of alkali metals or alkaline earth metals, C of alkali metals or alkaline earth metals
1-6Alkoxide, triethylamine, diisopropylethylamine, pyridine and N, N-dimethylaminopyridine or a plurality of alkoxides; preferably one or more of sodium hydroxide, potassium hydroxide, magnesium hydroxide, calcium hydroxide, sodium carbonate, potassium carbonate, magnesium carbonate, calcium carbonate, sodium methoxide, sodium ethoxide, magnesium methoxide, triethylamine, diisopropylethylamine, pyridine and N, N-dimethylaminopyridine; more preferably sodium hydroxide, potassium hydroxide, magnesium hydroxide, calcium hydroxide, sodium carbonate, potassium carbonate, magnesium carbonate, calcium carbonate, sodium methoxide, sodium ethoxide and mixtures thereof; sodium hydroxide, potassium hydroxide, calcium hydroxide and mixtures thereof are particularly preferred. Wherein the molar ratio of S-1 to the alkali is 1: 1-15, preferably 1:1-10, and more preferably 1: 1-8. The molar ratio of S-1 to dimethylamine or salts thereof is 1: 1-8, preferably 1: 1-5, wherein the dimethylamine salt is selected from inorganic salts of dimethylamine, preferably hydrochloride, bromate, sulfate and phosphate. The solvent is selected from polar solventAgent, preferably water, C
1-6Alcohol, C
1-6Ketones, C
1-6Ether, DMF, DMSO; more preferably water, C
1-6An alcohol; water, methanol, ethanol are particularly preferred; most preferably water. The reaction time is 0.2-3 h, preferably 0.5-1 h. The temperature is 0-60 deg.C, preferably 10-50 deg.C, more preferably 20-40 deg.C, and most preferably room temperature.
In the above step c, the hydrogenation catalyst is selected from palladium catalysts and platinum catalysts, preferably palladium catalysts in supported form; more preferably Pd/C, Pd (OH)
2/C, PdO/C, Mixed PdO-Pd (OH)
2/C、PdO/Al
2O
3Mixed PdO-Pd (OH)
2/Al
2O
3、PdO/SiO
2Mixed PdO-Pd (OH)
2/SiO
2、Pd/CaCO
3(ii) a Pd/C, Pd (OH) is particularly preferred
2/C, PdO/C; most preferably Pd/C; wherein the loading of palladium is in the range of 0.5% to 25%, preferably 0.5% to 25%, more preferably 1% to 20%, most preferably 5 to 15%; the mass ratio of the catalyst to the S-2 is 1: 4-50, preferably 1: 5-20, and more preferably 1: 5-10. The catalytic hydrogenation is carried out in the presence of hydrogen at a pressure of from atmospheric to 10 MPa. The temperature of the catalytic hydrogenation reaction is 0 to 80 deg.C, preferably 10 to 50 deg.C, more preferably 20 to 40 deg.C. The organic solvent is selected from C
1-6Alcohol, C
1-6Ethers or cyclic ethers, C
1-6Esters, C
1-6Ketones, C
1-6Halogenated alkanes, C
1-6One or more of alkyl nitrile and DMF, preferably one or more of ethyl acetate, dichloromethane, methanol, diethyl ether, tetrahydrofuran, DMF, ethanol, acetonitrile and acetone solution; more preferably two or more of ethyl acetate, dichloromethane, methanol, diethyl ether, tetrahydrofuran, DMF, ethanol, acetonitrile and acetone solution; particularly preferred are two of ethyl acetate, dichloromethane, methanol, diethyl ether, tetrahydrofuran, DMF, ethanol, acetonitrile and acetone solution, and most preferred is a mixed solvent of methanol and tetrahydrofuran, wherein the ratio of the two solvents is 0.1:1-10, preferably 0.5:1-5, more preferably 0.5:1-2, and most preferably 0.5:1, 1:1,1: 2.
In the step d, the reducing agent is a group containing metal hydride selected from sodium borohydride, lithium aluminum hydride and diisobutyl hydrideAluminum hydride, lithium tri-sec-butylborohydride, sodium bis (2-methoxyethoxy) aluminum hydride, sodium cyanoborohydride, trialkoxyaluminum hydride; preferably sodium borohydride, lithium aluminum hydride, diisobutylaluminum hydride, lithium tri-sec-butylborohydride; more preferred are sodium borohydride and lithium aluminum hydride. The molar ratio of reducing agent to S-3 is from 1 to 25:1, preferably from 5 to 20:1, more preferably from 6 to 15: 1. The reduction reaction may be carried out in the presence of a solvent selected from C
1-6Alcohol, C
1-6Ethers or cyclic ethers, C
1-6Esters, C
1-6One or more of alkyl nitrile or toluene; preferably one or more of methanol, ethanol, ethyl acetate, methyl tert-butyl ether, diethyl ether or tetrahydrofuran, more preferably methanol, ethanol or tetrahydrofuran. The reaction temperature is-20 ℃ to 80 ℃, preferably-10 ℃ to 50 ℃, more preferably 0 ℃ to 40 ℃. Step d is optionally performed under nitrogen or argon atmosphere.
Preferably, the reducing agent is dissolved in the solvent at 0 ℃ and added dropwise to the solvent containing S-3, the reaction system is heated under reflux for 3 to 8 hours, and then stirred at 25 ℃ to 40 ℃ for 8 to 16 hours.
In the step e, the methylating agent is selected from methyl iodide, methyl bromide, methyl chloride, methyl trifluoromethanesulfonate and dimethyl sulfate; preferably methyl iodide, methyl bromide; more preferably methyl iodide. The molar ratio of methylating agent to S-4 is from 1 to 30:1, preferably from 5 to 25:1, more preferably from 10 to 15: 1. The organic solvent is selected from C
1-6Alcohol, C
1-6Esters, C
1-6Ketones, C
1-6Ethers or cyclic ethers, C
1-6One or more of alkyl nitrile, DMF, or DMSO, preferably one or more of methanol, ethanol, ethyl acetate, acetone, methyl tert-butyl ether, diethyl ether, acetonitrile, or tetrahydrofuran, more preferably methanol, ethanol. The reaction temperature is 0 to 60 ℃, preferably 10 to 50 ℃, more preferably 20 to 40 ℃. Step e is optionally carried out under dark conditions; and e, optionally adding silver chloride after the reaction is completed to perform replacement to obtain the bufotoxin tryptamine quaternary ammonium salt (S-5).
The bufotoxin tryptamine quaternary ammonium salt can be a salt formed by methylation of the bufotoxin tryptamine, wherein an acid part is selected from: chloride ion, sulfate ion, phosphate ion, ascorbate, methanesulfonate, malate, succinate, glutarate, adipate, pimelate, suberate, etc.
In the aspect of pharmacological activity test, a large number of experiments show that the bufotenine quaternary ammonium salt has a strong analgesic effect. Screening and comparing various tryptamine components, finding that the toad venom tryptamine quaternary ammonium salt has the strongest analgesic effect, wherein the analgesic intensity of the toad venom tryptamine quaternary ammonium salt is 3 times that of the toad venom tryptamine; and the toad venom tryptamine quaternary ammonium salt still has obvious analgesic effect under the dosage that other tryptamine components in the toad venom are ineffective. And the toad venom tryptamine quaternary ammonium salt and low-dose morphine have the synergistic analgesic effect.
The toad tryptamine quaternary ammonium salt can regulate pain mediators derived from cyclooxygenase COX derived from arachidonic acid AA, has a regulating effect on pain mediators derived from lipoxygenase LOX, cytochrome CYP, linoleic acid LA and DHA, and is completely different from a mechanism of a non-sterene anti-inflammatory analgesic.
Therefore, the invention also relates to a pharmaceutical composition which comprises the toad venom tryptamine or the toad venom tryptamine quaternary ammonium salt with effective treatment amount and pharmaceutically acceptable auxiliary materials.
The invention also relates to a pharmaceutical composition which comprises the bufogenin or the bufogenin quaternary ammonium salt and opioid such as morphine hydrochloride.
The invention also relates to application of the bufotamine or the bufotamine quaternary ammonium salt in preparation of analgesic or anti-inflammatory drugs.
Preferably, the bufotenine quaternary ammonium salt is 2- (5-hydroxy-1H-indol-3-yl) -N, N, N-trimethylethan-1-amine chloride.
Has the advantages that: compared with the prior art, the invention has the following advantages:
compared with the prior art, the method has the advantages of simple steps, safe and convenient operation, high reaction efficiency, easy purification and separation, mild conditions, low cost, total reaction yields of the bufotoxin tryptamine and the quaternary ammonium salt thereof of more than 65 percent and 60 percent respectively, high purity (> 99.0%) of the synthesized product, and suitability for industrial production.
According to the invention, a large number of experiments are carried out to analyze and research the analgesic effect of the bufotenine, the serotonin and the bufotenine quaternary ammonium salt, and the result shows that the bufotenine quaternary ammonium salt has the strongest analgesic effect, and the bufotenine quaternary ammonium salt still has the obvious analgesic effect under the ineffective dose of other tryptamine components. And the toad venom tryptamine quaternary ammonium salt and the morphine have the synergistic analgesic effect.
Drawings
Fig. 1 is a schematic structural diagram of bufotenine and quaternary ammonium salt thereof.
Fig. 2 is a reaction flow chart of the method for synthesizing bufotenine and quaternary ammonium salt thereof.
Figure 3 is a graph of the effect of bufotenine on the hind paw thermal pain threshold of mice.
FIG. 4 is a graph of the effect of bufotenine on the mechanical pain threshold of the right hind paw of a mouse.
Fig. 5 is a graph of the behavioral effects of bufotenine on formalin-induced pain model mice.
Figure 6 is a graph of the effect of bufotamine quaternary on the hind paw thermal pain threshold in mice.
FIG. 7 is a graph showing the effect of bufotamine quaternary ammonium salt on the mechanical pain threshold of the right hind paw of a mouse
Fig. 8 is a graph of the behavioral effects of bufotamine quaternary ammonium on formalin-induced pain model mice.
Detailed Description
The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the specific material ratios, process conditions and results thereof described in the examples are illustrative only and should not be taken as limiting the invention as detailed in the claims.
Example 1
Preparation of 2- (5- (benzyloxy) -1H-indol-3-yl) -2-oxoacetyl chloride (S-1)
At room temperature, a 100mL round-bottom flask is taken, 2.7mL of anhydrous ether and oxalyl chloride (175 mu L,2.0mmol) are sequentially added, after the mixture is fully and uniformly stirred, 5-benzyloxy indole (220mg,1.0mmol) is added into the flask, after the mixture is fully stirred for 30 minutes, reaction liquid is filtered by suction, and a filter cake is repeatedly washed by ether, so 302.9mg of reddish brown solid precipitate, namely S-1 crude product, is obtained, and the yield is 95%.
Preparation of 2- (5- (benzyloxy) -1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide (S-2)
At room temperature, a 100mL round-bottom flask is taken, potassium hydroxide (360mg,6.38mmol) and 4.15mL of water and dimethylamine hydrochloride (350mg,4.25mmol) are sequentially added into the round-bottom flask, after uniform mixing, S-1(310mg,1.0mmol) is slowly added into the mixed solution, after stirring for 40 minutes, suction filtration and washing with anhydrous ether are carried out, a white solid is obtained, namely an S-2 crude product, the crude product is directly stirred with silica gel after drying, column chromatography separation is carried out, pure ethyl acetate is taken as an eluent, a pure product of S-2 is obtained, about 287.1mg is a white solid, and the yield is 90%.
The structural analytic formula is:
1H NMR(500MHz,CDCl
3)δ8.96(s,1H),7.99(d,J＝2.1Hz,1H),7.91(d,J＝3.2Hz,1H),7.52(d,J＝7.4Hz,1H),7.42(t,J＝7.5Hz,1H),7.35(dd,J＝13.1,8.1Hz,1H),7.05(dd,J＝8.8,2.5Hz,1H),5.18(s,2H),3.12(s,2H),3.10(s,2H).
preparation of 2- (5-hydroxy-1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide (S-3)
At room temperature, a three-necked flask was taken and heated in the presence of a methanol-tetrahydrofuran ═ 1:2, preparing solution, taking 600 mu L of each solution, adding S-2(50mg,0.155mol) and Pd/C (10%, 5mg), filling hydrogen, reacting for 18 hours at normal temperature and normal pressure, after the reaction is completed, leaching with diatomite, removing the catalyst, repeatedly washing with ethyl acetate, drying the filtrate with anhydrous sodium sulfate, mixing the sample with silica gel, and performing column chromatography separation with dichloromethane: methanol 20:1 as eluent, to give S-3 as a pure product, about 33.5mg, as an oil, in 93% yield.
The structural analytic formula is:
1H NMR(500MHz,DMSO)δ12.07(s,1H),9.17(s,1H),7.96(d,J＝3.2Hz,1H),7.50(s,1H),7.32(d,J＝8.7Hz,1H),6.76(dd,J＝8.7,2.3Hz,1H),2.98(s,3H),2.91(s,3H).
preparation of 2- (2- (dimethylamino) ethyl) -1H-indol-5-ol (S-4; bufotenine)
Dissolving S-3(35mg,0.11mmol) in a round-bottom flask containing tetrahydrofuran (2mL) at 0 ℃, uniformly mixing sodium borohydride (42mg,1.1mmol) and tetrahydrofuran (1.5mL), slowly dropwise adding the mixture into the flask, cooling and refluxing for 5 hours under the protection of nitrogen, stirring for 8 hours at 40 ℃, cooling in an ice bath after the reaction is finished, adding sodium hydroxide to quench the reaction, stirring for 30 minutes, extracting, repeatedly extracting with ethyl acetate for three times, taking an organic layer, drying with anhydrous sodium sulfate, mixing with silica gel, performing column chromatography separation, and performing column chromatography with dichloromethane: methanol 10:1 as eluent, to obtain a pure product of S-4 (bufotamine), about 25.9mg, as a tan oil, with a yield of 84%.
The structural analytic formula is:
1H NMR(500MHz,DMSO)δ10.49(s,1H),8.65(s,1H),7.12(d,J＝8.6Hz,1H),7.04(d,J＝2.1Hz,1H),6.81(d,J＝2.2Hz,1H),6.59(dd,J＝8.6,2.3Hz,1H),2.80–2.71(m,2H),2.62–2.54(m,2H),2.29(s,7H).
preparation of 2- (5-hydroxy-1H-indol-3-yl) -N, N, N-trimethylethan-1-amine chloride (S-5)
Taking S-4(50mg,0.25mmol) in a round-bottom flask containing methanol (5ml) at room temperature in a dark place, adding methyl iodide (1.9ml,3.0mmol) into the round-bottom flask containing the methanol (5ml), uniformly mixing, reacting for 48h, performing suction filtration, repeatedly washing the methanol, taking filter residues, dissolving the filter residues in the round-bottom flask containing the methanol, adding excessive AgCl, reacting in the dark place, performing suction filtration after complete reaction, taking the filter residues, mixing the filter residues with silica gel, performing column chromatography separation, and performing column chromatography separation by using dichloromethane: methanol 5:1 as eluent, to give S-5 as a pure product, about 48.3mg, as a tan oil, yield 90%.
The structural analytic formula is:
1H NMR(500MHz,DMSO)δ10.49(s,1H),7.12(d,J＝8.6Hz,1H),7.04(d,J＝2.1Hz,1H),6.81(d,J＝2.2Hz,1H),6.59(dd,J＝8.6,2.3Hz,1H),2.80–2.71(m,2H),2.62–2.55(m,2H),2.29(s,6H).
example 2
Preparation of 2- (5- (benzyloxy) -1H-indol-3-yl) -2-oxoacetyl chloride (S-1)
At room temperature, a 100mL round-bottom flask was taken, 2.7mL of anhydrous ether and oxalyl chloride (175. mu.L, 2.0mmol) were sequentially added, and after stirring sufficiently and uniformly, 5-benzyloxyindole (220mg,1.0mmol) was added to the flask, and after stirring sufficiently and for 30 minutes, the reaction solution was suction filtered, and the filter cake was washed repeatedly with ether, to obtain 299.8mg of a reddish brown solid precipitate as a crude S-1 product, with a yield of 94%.
Preparation of 2- (5- (benzyloxy) -1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide (S-2)
At room temperature, a 100mL round-bottom flask is taken, sodium hydroxide (170mg,4.25mmol) and 4.15mL water and dimethylamine hydrochloride (350mg,4.25mmol) are sequentially added into the round-bottom flask, after uniform mixing, S-1(310mg,1.0mmol) is slowly added into the mixed solution, after stirring for 40 minutes, suction filtration and washing with anhydrous ether are carried out, a white solid is obtained, namely an S-2 crude product, after drying, direct silica gel stirring is carried out, column chromatography separation is carried out, pure ethyl acetate is taken as an eluent, a pure product of S-2 is obtained, about 290.2mg is a white solid, and the yield is 91%.
The structural analytic formula is:
1H NMR(500MHz,CDCl
3)δ8.96(s,1H),7.99(d,J＝2.1Hz,1H),7.91(d,J＝3.2Hz,1H),7.52(d,J＝7.4Hz,1H),7.42(t,J＝7.5Hz,1H),7.35(dd,J＝13.1,8.1Hz,1H),7.05(dd,J＝8.8,2.5Hz,1H),5.18(s,2H),3.12(s,2H),3.10(s,2H)。
preparation of 2- (5-hydroxy-1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide (S-3)
At room temperature, a three-necked flask was taken and heated in the presence of a methanol-tetrahydrofuran ═ 1:1, preparing solutions, taking 700mL of each solution, adding S-2(50mg,0.155mol) and Pd/C (10%, 5mg), filling hydrogen, reacting for 22 hours at normal temperature and normal pressure, after the reaction is completed, leaching with diatomite, removing a catalyst, repeatedly washing with ethyl acetate, drying filtrate with anhydrous sodium sulfate, mixing silica gel with a sample, and performing column chromatography separation with dichloromethane: methanol 20:1 as eluent, to give S-3 as a pure product, about 34.1mg, as an oil, in 94% yield.
The structural analytic formula is:
1H NMR(500MHz,DMSO)δ12.07(s,1H),9.17(s,1H),7.96(d,J＝3.2Hz,1H),7.50(s,1H),7.32(d,J＝8.7Hz,1H),6.76(dd,J＝8.7,2.3Hz,1H),2.98(s,3H),2.91(s,3H).
preparation of 2- (2- (dimethylamino) ethyl) -1H-indol-5-ol (S-4; bufotenine)
Dissolving S-3(35mg,0.11mmol) in a round-bottom flask containing tetrahydrofuran (2mL) at 0 ℃, uniformly mixing lithium aluminum hydride (37mg,1.1mmol) and tetrahydrofuran (1.5mL), slowly dropwise adding the mixture into the flask, cooling and refluxing for 4 hours under the protection of nitrogen, stirring for 10 hours at 40 ℃, cooling in an ice bath after the reaction is finished, adding sodium hydroxide to quench the reaction, stirring for 30 minutes, extracting, repeatedly extracting with ethyl acetate for three times, taking an organic layer, drying with anhydrous sodium sulfate, mixing with silica gel, performing column chromatography separation, and performing column chromatography with dichloromethane: methanol 10:1 was used as the eluent to give S-4 (bufotenine) as a pure product, about 26.2mg, as a tan oil in 85% yield.
The structural analytic formula is:
1H NMR(500MHz,DMSO)δ10.49(s,1H),8.65(s,1H),7.12(d,J＝8.6Hz,1H),7.04(d,J＝2.1Hz,1H),6.81(d,J＝2.2Hz,1H),6.59(dd,J＝8.6,2.3Hz,1H),2.80–2.71(m,2H),2.62–2.54(m,2H),2.29(s,7H).
preparation of 2- (5-hydroxy-1H-indol-3-yl) -N, N, N-trimethylethan-1-amine chloride (S-5)
Taking S-4(50mg,0.25mmol) in a round-bottom flask containing methanol (5ml) at room temperature in a dark place, adding methyl iodide (2.2ml,3.5mmol) into the round-bottom flask containing the methanol (5ml), uniformly mixing, reacting for 48h, performing suction filtration, repeatedly washing the methanol, taking filter residues, dissolving the filter residues in the round-bottom flask containing the methanol, adding excessive AgCl, reacting in the dark place, performing suction filtration after complete reaction, taking the filter residues, mixing the filter residues with silica gel, performing column chromatography separation, and performing column chromatography separation by using dichloromethane: methanol 5:1 as eluent, to give S-5 as a pure product, about 40.8mg, as a tan oil, 91% yield.
The structural analytic formula is:
1H NMR(500MHz,DMSO)δ10.49(s,1H),7.12(d,J＝8.6Hz,1H),7.04(d,J＝2.1Hz,1H),6.81(d,J＝2.2Hz,1H),6.59(dd,J＝8.6,2.3Hz,1H),2.80–2.71(m,2H),2.62–2.55(m,2H),2.29(s,6H).
example 3
Preparation of 2- (5- (benzyloxy) -1H-indol-3-yl) -2-oxoacetyl chloride (S-1)
At room temperature, a 100mL round-bottom flask was taken, 2.7mL of anhydrous ether and oxalyl chloride (175. mu.L, 2.0mmol) were sequentially added, and after stirring sufficiently and uniformly, 5-benzyloxyindole (220mg,1.0mmol) was added to the flask, and after stirring sufficiently and 40 minutes, the reaction solution was suction filtered, and the filter cake was washed repeatedly with ether, to obtain 293.1mg of a reddish brown solid precipitate as a crude S-1 product, with a yield of 95%.
Preparation of 2- (5- (benzyloxy) -1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide (S-2)
At room temperature, a 100mL round-bottom flask is taken, sodium hydroxide (255mg,6.38mmol) and 4.15mL of water and dimethylamine hydrochloride (350mg,4.25mmol) are sequentially added into the round-bottom flask, after uniform mixing, S-1(310mg,1.0mmol) is slowly added into the mixed solution, after stirring for 40 minutes, suction filtration and washing with anhydrous ether are carried out, a white solid is obtained, namely an S-2 crude product, after drying, direct silica gel stirring is carried out, column chromatography separation is carried out, pure ethyl acetate is taken as an eluent, a pure product of S-2 is obtained, about 293.5mg is a white solid, and the yield is 92%.
The structural analytic formula is:
1H NMR(500MHz,CDCl
3)δ8.96(s,1H),7.99(d,J＝2.1Hz,1H),7.91(d,J＝3.2Hz,1H),7.52(d,J＝7.4Hz,1H),7.42(t,J＝7.5Hz,1H),7.35(dd,J＝13.1,8.1Hz,1H),7.05(dd,J＝8.8,2.5Hz,1H),5.18(s,2H),3.12(s,2H),3.10(s,2H).
preparation of 2- (5-hydroxy-1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide (S-3)
At room temperature, a three-necked flask was taken and heated to room temperature under the following conditions: 1, preparing solutions, taking 700 mu L of each solution, adding S-2(50mg,0.155mol) and Pd/C (10%, 5mg), filling hydrogen, reacting for 20 hours at normal temperature and normal pressure, after the reaction is completed, leaching with diatomite, removing a catalyst, repeatedly washing with ethyl acetate, drying filtrate with anhydrous sodium sulfate, mixing silica gel with a sample, performing column chromatography separation, and performing column chromatography separation with dichloromethane: methanol 20:1 as eluent, to give S-3 as a pure product, about 33.1mg, as an oil, in 92% yield.
The structural analytic formula is:
1H NMR(500MHz,DMSO)δ12.07(s,1H),9.17(s,1H),7.96(d,J＝3.2Hz,1H),7.50(s,1H),7.32(d,J＝8.7Hz,1H),6.76(dd,J＝8.7,2.3Hz,1H),2.98(s,3H),2.91(s,3H).
preparation of 2- (2- (dimethylamino) ethyl) -1H-indol-5-ol (S-4; bufotenine)
Dissolving S-3(35mg,0.11mmol) in a round-bottom flask containing tetrahydrofuran (2mL) at 0 ℃, uniformly mixing lithium aluminum hydride (55.5mg,1.65mmol) and tetrahydrofuran (2.0mL), slowly dropwise adding into the flask, cooling and refluxing for 5 hours under the protection of nitrogen, stirring for 10 hours at 40 ℃, cooling in an ice bath after the reaction is finished, adding sodium hydroxide to quench the reaction, stirring for 1 hour, extracting, repeatedly extracting with ethyl acetate for three times, taking an organic layer, drying with anhydrous sodium sulfate, mixing with silica gel, performing column chromatography separation, separating with dichloromethane: methanol 10:1 as eluent, to obtain a pure product of S-4 (bufotamine), about 23.7mg, as a tan oil, with a yield of 84%.
The structural analytic formula is:
1H NMR(500MHz,DMSO)δ10.49(s,1H),8.65(s,1H),7.12(d,J＝8.6Hz,1H),7.04(d,J＝2.1Hz,1H),6.81(d,J＝2.2Hz,1H),6.59(dd,J＝8.6,2.3Hz,1H),2.80–2.71(m,2H),2.62–2.54(m,2H),2.29(s,7H).
preparation of 2- (5-hydroxy-1H-indol-3-yl) -N, N, N-trimethylethan-1-amine chloride (S-5)
Taking S-4(50mg,0.25mmol) in a round-bottom flask containing methanol (5ml) at room temperature in a dark place, adding methyl iodide (1.9ml,3.0mmol) into the round-bottom flask containing the methanol (5ml), uniformly mixing, reacting for 48h, performing suction filtration, repeatedly washing the methanol, taking filter residues, dissolving the filter residues in the round-bottom flask containing the methanol, adding excessive AgCl, reacting in the dark place, performing suction filtration after complete reaction, taking the filter residues, mixing the filter residues with silica gel, performing column chromatography separation, and performing column chromatography separation by using dichloromethane: methanol 5:1 as eluent, to give S-5 as a pure product, about 48.8mg, as a tan oil, 91% yield.
The structural analytic formula is:
1H NMR(500MHz,DMSO)δ10.49(s,1H),7.12(d,J＝8.6Hz,1H),7.04(d,J＝2.1Hz,1H),6.81(d,J＝2.2Hz,1H),6.59(dd,J＝8.6,2.3Hz,1H),2.80–2.71(m,2H),2.62–2.55(m,2H),2.29(s,6H).
activity measurement examples
Unless otherwise indicated, the quaternary ammonium salt of bufotenine used in the activity assay process of the present invention is 2- (5-hydroxy-1H-indol-3-yl) -N, N-trimethylethan-1-aminium chloride.
Example 4
1. Evaluation of analgesic effect of bufotenine by hot plate method
1.1 materials and methods
1.1.1 animals: healthy SPF grade adult ICR mice, female, having a weight of 18-22g and a age of 6-8 weeks, were provided by Qinglongshan laboratory animal farms, Nanjing. Before the experiment, all mice are adaptively raised in the animal room of the laboratory for 3-4 days, the room temperature is maintained at 20-25 ℃, the relative humidity is 40% -60%, the animals are raised in cages during the experiment, the light-dark period is 12h/12h, the animals can freely eat and drink water, and the food is standard granular feed. The experimental procedures strictly adhere to national institute approved guidelines for the care and use of laboratory animals.
1.1.2 apparatus: a numerical control super constant temperature tank, an electronic balance, a 1mL injector and a cage;
1.1.3 drugs: diluting bufotoxin tryptamine with 0.9% sodium chloride during injection to prepare a solution of 0.5mg/mL and 1.5 mg/mL; morphine hydrochloride injection is diluted by 0.9 percent sodium chloride during injection to prepare a solution of 0.025 mg/mL; 0.9% sodium chloride solution.
1.2 Experimental methods
1.2.1 preselecting animals
Mice with a painful response within 30s were pre-selected. The mice are placed in a hot plate instrument, the temperature is set to be 55 +/-0.5 ℃, the pain threshold value of the mice is determined, the pain threshold value is measured for 2 times, the pain threshold value is measured for 1 time every 5min, and the hindpaw of the mice is used as an observation index. If the mice do not lick the hind paw, escape, jump within 30s, they are discarded. The time for licking the feet for the first time before the drug is taken as the pain threshold value.
1.2.2 grouping and administration
Selecting pre-selected qualified mice, and dividing into normal saline group, morphine hydrochloride injection group, and bufogenin group (high and low dose), each group containing 12 mice. Each group of mice was then dosed i.p. at 0.1mL/10g, with dosing information given in table 1 below.
2.2.3 thermal threshold measurement method
Before drug intervention, the temperature of a hot plate is adjusted to 55 ℃, the mouse is stabilized for one hour, the mouse is placed in a hot plate instrument 30min, 60min, 90min and 120min after administration, the first time of foot licking reaction time, namely a pain threshold (HPPT), is observed, and if the mouse has no pain reaction within 60s, the mouse is immediately taken out and calculated according to 60 s.
Percent increase in pain threshold (post-dose pain threshold-pre-dose pain threshold)/pre-dose pain threshold × 100%
2.2.4 statistical treatment
Statistical analysis and mapping are carried out on the experimental data by adopting statistical analysis software Excel 2016 and GraphPad Prism 8 software. Data are presented as mean ± SEM, T-test for metrology data of two independent samples. P <0.05 indicates that the difference is statistically significant.
TABLE 1 administration information table for different groups of mice
| Group of | n | Intervention drug | Dosage form | Equivalent dose for human body |
| Physiological saline group | 12 | Physiological saline | - | - |
| Morphine hydrochloride group | 12 | Morphine hydrochloride | 0.25mg/kg | 0.002mg/kg |
| Low dose group of bufotenine | 12 | Toad venom tryptamine | 5mg/kg | 0.41mg/kg |
| High dose group of bufotenine | 12 | Toad venom tryptamine | 15mg/kg | 1.22mg/kg |
2.3 Experimental results and discussion
2.3.1 Effect of bufotenine on the Hot pain threshold of the hind paw of mice
The effect of bufotenine on the hind paw thermal pain threshold of mice is shown in table 2 below and fig. 2. The results show that compared with the normal saline group, the pain threshold value of the bufotamine high-dose group (15mg/kg) is obviously improved (P is less than 0.05) after administration, a certain analgesic effect is shown, compared with the group before administration, the analgesic effect reaches the highest in 60 minutes, and the pain threshold value improvement rate is 34.92%. The low dose group of bufotenine (5mg/kg) showed no analgesic effect.
TABLE 2 Effect of bufotenine on the hind paw thermal pain threshold in mice: (
n＝10-12)
Note: p is less than or equal to 0.01, and has extremely obvious difference compared with the normal saline; p is less than or equal to 0.05, and the difference is obvious compared with the normal saline.
##P is less than or equal to 0.01, and has very obvious difference compared with the prior medicine;
#p is less than or equal to 0.05, and is obviously different from the medicine before administration.
2. Evaluation of analgesic effect of formalin-induced pain model on toad venom tryptamine
2.1 materials and methods
2.1.1 animals: healthy SPF grade adult ICR mice, half male and female, 18-22g in weight, 6-8 weeks old, were provided by Qinglongshan laboratory animal farms, Nanjing. Before the experiment, all mice are adaptively raised in the animal room of the laboratory for 3-4 days, the room temperature is maintained at 20-25 ℃, the relative humidity is 40% -60%, the animals are raised in cages during the experiment, the light-dark period is 12h/12h, the animals can freely eat and drink water, and the food is standard granular feed. The experimental procedures were strictly in accordance with national institute approved guidelines for the care and use of laboratory animals.
2.1.2 apparatus: a Von Frey fiber stimulating needle, a cage, an electronic balance, a 1mL syringe, a video device and the like;
2.1.3 drugs: diluting bufotenine with 0.9% sodium chloride during injection to obtain 0.5mg/mL and 1.5mg/mL solutions; morphine hydrochloride injection is diluted by 0.9 percent sodium chloride during injection to prepare a solution of 0.025 mg/mL; 0.9% sodium chloride solution.
2.2 Experimental methods
2.2.1 grouping and administration
Healthy ICR mice were selected and randomly divided into model groups, morphine hydrochloride injection groups, and bufotoxin tryptamine groups (high and low doses) according to body weight. Each group of mice was then dosed i.p. at 0.1mL/10g, with dosing information as shown in table 3. After 15min post-administration, 20 μ L of 2.5% formalin (0.925% formaldehyde) solution was administered subcutaneously (s.c.) along the right foot of the mouse, and the animal was observed for behavioral performance by mechanical pain threshold determination.
2.2.2 mechanical pain threshold detection method
The threshold value of the foot contraction response of the right hind paw of the mouse to the mechanical stimulation, namely the mechanical foot contraction threshold value (PWMT), is measured by von Frey cellosilk, and the mouse is placed in a mouse cage which is about 30cm higher than the experimental table surface and is in a quiet state after being adapted for 5-10 min. During measurement, the fiber filaments representing different stimulation forces are used for vertically stimulating the right hind paw of the mouse from low to high in sequence, the force is slowly exerted until the fiber filaments are bent and angled for 1.5-2s, and the stimulation is finished. And (3) observing that any behavior of the mouse of hoarse, licking, flicking and bouncing legs is recorded as positive force, selecting the next type of fiber silk with the force lower than the force to continue stimulation until the low-force fiber silk is stimulated negatively, recording the last value of the last low-force fiber silk as the mechanical foot-retracting threshold value of the last low-force fiber silk, and recording the stimulation force represented by the fiber silk used at the moment. If the intensity is negative, selecting the fiber silk with the intensity larger than the intensity to stimulate until the intensity is positive, and recording the intensity value at the moment as the pain threshold value. Mechanical pain thresholds were recorded 15min, 30min, 45min, 60min, 75min and 90min after formalin administration in mice, respectively.
2.2.3 behavioral observations of pain
The right paw of the mice was observed for pain behavioural performance given 2.5% formalin solution. Mice given 2.5% formalin solution to the right paw were placed in a cage with a flat bottom surface, and the time for the right paw to leave the bottom surface of the cage (total time for licking, contracting and lifting the paw) after stimulation was observed for a total of 45min, and the total time for lifting the paw was recorded every 5 min.
TABLE 3 administration information table for different groups of mice
| Group of | n | Intervention drug | Dosage form | Equivalent dose for human body |
| Model set | 10 | Physiological saline | - | - |
| |
10 | Morphine hydrochloride | 0.25mg/kg | 0.002mg/kg |
| Low dose group of |
10 | Toad venom tryptamine | 5mg/kg | 0.41mg/kg |
| High dose group of |
10 | Toad venom tryptamine | 15mg/kg | 1.22mg/kg |
2.3 results of the experiment
2.3.1 Effect of bufotenine on the mechanical pain threshold of the right hind paw of mice
The results of the effects of the bufotenine on the mechanical pain threshold of the right hind paw of the mouse are shown in the following table 4 and fig. 3, and the results show that the mechanical pain threshold of the right paw of the bufotenine low-dose group (5mg/kg) is slightly increased (P is less than 0.05) and the mechanical pain threshold of the right paw of the high-dose group (15mg/kg) is significantly increased (P is less than 0.01) within 90 minutes after the administration of formalin, and the maximum mechanical pain threshold is reached in 60 minutes and is (1.32 +/-1.23) g and (2.1 +/-1.38) g respectively.
Table 4 effect of bufotenine on mechanical pain threshold of right hind paw of mouse (X ± SEM, n ═ 10)
Note: p is less than or equal to 0.01, and has extremely obvious difference compared with a model group; p is less than or equal to 0.05, and is significantly different from the model group.
2.3.2 Effect of bufotenine on mouse behaviourology in formalin-induced pain model
The formalin model of foot swelling and pain shows a biphasic response, one phase (0-5 min; first phase) being the direct stimulation of peripheral nociceptors by formalin, and two phases (15-45 min; second phase) being the central neuron sensitization, with a relatively short resting period (5-10 min) in between. The nociceptive behaviour of formalin was quantified in this experiment as the length of time it took to lick or bite the injection paw within 45 minutes (divided into 9 cycles of 5 minutes each). The behavioral results of the bufotenine compounds on formalin-induced pain model mice are shown in the following table 5 and fig. 4. The results show that the low and high dose bufotamine (5mg/kg and 15mg/kg) groups significantly inhibited mice from licking or biting the injected feet (P < 0.01) compared to the normal saline group within 45 minutes after formalin administration.
TABLE 5 Effect of bufotenine on formalin nociceptin model mouse behaviourology (X + -SEM, n ═ 6)
Note: p is less than or equal to 0.01, and has extremely obvious difference compared with a model group; p is less than or equal to 0.05, and is significantly different from the model group.
Example 5
1. Evaluation of analgesic effect of toad venom tryptamine quaternary ammonium salt by hot plate method
1.1 materials and methods
1.2 Experimental methods are as before
TABLE 6 administration information table for different groups of mice
| Group of | n | Dosage form | Equivalent dose for human body |
| Physiological saline group | 12 | - | - |
| Morphine hydrochloride group | 12 | 0.25mg/kg | 0.002mg/kg |
| Toad venom tryptamine quaternary ammonium salt low-dose group | 12 | 5mg/kg | 0.41mg/kg |
| High-dose group of toad venom tryptamine quaternary ammonium salt | 12 | 15mg/kg | 1.22mg/kg |
1.3. Experimental results and discussion
Effect of bufotenine Quaternary ammonium salt on the hind paw thermal pain threshold of mice
The effect of bufotamine quaternary ammonium salts on the hind paw thermal pain threshold in mice is shown in table 7 below and figure 6. The results show that compared with the normal saline group, the pain threshold value of the low-high dose group (5mg/kg and 15mg/kg) of the bufotamine quaternary ammonium salt is obviously improved (P is less than 0.05) after administration, a certain analgesic effect is shown, compared with the group before administration, the analgesic effect reaches the highest in 60 minutes, and the pain threshold value improvement rates are respectively 29.61% and 60.62% (P is less than 0.01).
TABLE 7 Effect of bufotenine Quaternary ammonium salt on the hind paw thermal pain threshold in mice: (
n＝10-12)
Note: p is less than or equal to 0.01, and has extremely obvious difference compared with the normal saline; p is less than or equal to 0.05, and the difference is obvious compared with the normal saline.
##P is less than or equal to 0.01, and has very obvious difference compared with the prior medicine;
#p is less than or equal to 0.05, and is obviously different from the medicine before administration.
2. Evaluation of analgesic effect of formalin-induced pain model on toad venom tryptamine quaternary ammonium salt
2.1 materials and methods
TABLE 8 administration information table for different groups of mice
| Group of | n | Dosage form | Equivalent dose for human body |
| Model set | 10 | - | - |
| |
10 | 0.25mg/kg | 0.002mg/kg |
| Toad venom tryptamine quaternary ammonium salt low- |
10 | 5mg/kg | 0.41mg/kg |
| High-dose group of toad venom tryptamine quaternary |
10 | 15mg/kg | 1.22mg/kg |
2.2 results of the experiment
Effect of bufotenine quaternary ammonium salt on mechanical pain threshold of right hind paw of mouse
The effect of the bufotamine quaternary ammonium salt on the mechanical pain threshold of the right hind paw of the mouse is shown in the following table 9 and fig. 7, and the results show that the mechanical pain threshold of the right paw of the mouse is obviously increased in the low-high dose group (5mg/kg and 15mg/kg) of the bufotamine quaternary ammonium salt and the model group (P is less than 0.01, the maximum mechanical pain threshold is reached in 60 minutes, and is respectively (2.24 +/-1.45) g and (9.04 +/-3.13) g (P is less than 0.05, and P is less than 0.010) within 90 minutes after formalin is administered.
TABLE 9 Effect of bufotenine quaternary ammonium salts on the mechanical pain threshold of the right hind paw in mice (X + -SEM, n ═ 10)
Note: p is less than or equal to 0.01, and has extremely obvious difference compared with a model group; p is less than or equal to 0.05, and is significantly different from the model group.
2.3 Effect of bufotamine Quaternary ammonium salt on mouse behaviourology in formalin-induced pain model
The formalin model of foot swelling and pain shows a biphasic response, one phase (0-5 min; first phase) being the direct stimulation of peripheral nociceptors by formalin, and two phases (15-45 min; second phase) being the central neuron sensitization, with a relatively short resting period (5-10 min) in between. The nociceptive behaviour of formalin was quantified in this experiment as the length of time it took to lick or bite the injection paw within 45 minutes (divided into 9 cycles of 5 minutes each). The behavioral results of bufotenine quaternary ammonium salt on formalin-induced pain model mice are shown in table 10 and fig. 8. The results show that the low and high dose groups (5mg/kg and 15mg/kg) of the bufotamine quaternary ammonium salt significantly inhibit the mice from licking or biting the injected feet (P < 0.01) compared with the normal saline group within 45 minutes after formalin administration.
TABLE 10 Effect of bufotenine quaternary ammonium salts on the behavioral analysis of formalin nociceptin model mice (X + -SEM, n ═ 6)
Note: p is less than or equal to 0.01, and has extremely obvious difference compared with a model group; p is less than or equal to 0.05, and is significantly different from the model group.
3. Influence of toad tryptamine quaternary ammonium salt on foot inflammation medium of formalin-induced pain model mouse
3.1 materials and methods
3.1.1 reagents
N-hexane (guo pharmaceutical group chemical reagent ltd, analytical grade), ethyl acetate (nanjing chemical reagent ltd, analytical grade), sodium chloride (guo pharmaceutical group chemical reagent ltd, analytical grade), acetonitrile (TEDIA reagent ltd, chromatographic grade), methanol (TEDIA reagent ltd, chromatographic grade), isopropanol (MERCK company, germany, chromatographic grade), formic Acid (Sigma company, chromatographic grade), Arachidonic Acid (available from Cayman).
3.1.2 instruments
AB SCIEX QTRAP 5500 mass spectrometer (USA AB), SHIMADZU LC-20AD XR ultra high performance liquid chromatograph (Shimadzu corporation), EPED ultra pure water instrument (Nanjing Yipu Yi Da science development Co., Ltd.), FA2004N analytical balance (Shanghai precision science instruments Co., Ltd.), ZLS-1 vacuum centrifugal concentrator (Hunan Hexi instruments and Equipment Co., Ltd.), KH3200B ultrasonic cleaner (Kunshan Seawa ultrasonic instruments Co., Ltd.), SHZ-D (III) circulating water type vacuum pump (Henan province Zhua instruments Co., Ltd.), XW-80A vortex mixer (Haimax Linbel instruments manufacturing Co., Ltd.), D3024R low temperature high speed centrifuge (American SCILOGEX corporation), IMS-20 ice maker (Hecheng Xuekao appliances Co., Ltd.).
3.2 Experimental methods
3.2.1 preparation of reagents
Preparation of 0.9% physiological saline: precisely weighing 0.9g of NaCl, adding 100mL of ultrapure water, uniformly stirring by using a glass rod, and standing at 4 ℃ for more than 30 min.
Preparing an extracting solution: preparing an extracting solution containing n-hexane and ethyl acetate according to a v/v-1: 1, and storing at-80 ℃ for more than 30 min.
Preparing a complex solution: 4.5mL of acetonitrile, 5mL of isopropanol and 0.5mL of ultrapure water were precisely aspirated to prepare 10mL of an isopropanol-acetonitrile-water solution. The AA-d8 solution with the mother liquor concentration of 10mg/mL is diluted by the solution to prepare the isopropanol-acetonitrile-water solution (compound solution) containing 200ng/mLAA-d 8.
3.2.2 sample handling
Placing the right hind paw of the mouse on an ice bag, shearing the paw with scissors, transferring the paw into a glass homogenizer, adding pre-cooled 0.9% physiological saline with the weight 4 times that of the paw, and inserting the glass homogenizer into an ice box for grinding until the paw is in a homogenate state. Transferring the homogenized tissues into a glass test tube, adding 8 times of full-weight precooled extracting solution, uniformly mixing for 1min in a vortex mode, carrying out ultrasonic extraction twice in ice-water bath for 20min each time, taking the upper layer liquid, centrifuging for 15min under the condition of 4000r/min, taking the supernatant, concentrating and volatilizing at 37 ℃, adding 8 times of full-weight precooled extracting solution into the lower layer sediment again, repeating the operation, combining the centrifuged supernatant with a corresponding tube sample, concentrating and volatilizing again. Precisely sucking 500 μ L of redissolution for redissolving, passing through a membrane, centrifuging at 4 deg.C and 14000r/min for 30min, and sampling the supernatant.
3.2.3 chromatographic conditions
The chromatographic column is an XBridge C18 chromatographic column (4.6mm multiplied by 100mm, 3.5 μm), the volume flow rate is 0.7mL/min, the sample injection amount is 10 μ L, and the column temperature is 50 ℃. Mobile phase a (water: acetonitrile: formic acid 70: 30: 0.02), mobile phase B (acetonitrile: isopropanol 50: 50), gradient elution procedure: 0-3 min, 25% B; 3-11 min, 45% B; 11-13 min, 60% B; 13-18 min, 75% B; for 18-18.5 min, 90% B; 20-21 min, 0% B; 21-25 min, 0% B.
Mass spectrum conditions: electrospray ion source (ESI); scanning mode: anion mode, mass spectrum parameters CUR 10psi, GSI 30psi, GS2 30psi, IS-4500V, CAD HIGH, TEMP 525, IHE ON, XP-10V.
3.2.4 data processing
Adopting Analyst 5.2 software of AB company to derive sample data, analyzing with GraphPad Prism 8 software, performing t test statistical processing with Excel, and adopting result data
It is shown that P <0.05 is significant, and P < 0.01 is significant. The specific calculation formula is as follows: n is Ax/Ai/Mx V20 (Ax: peak area of each substance in the sample, Ai: peak area of internal standard AA-d8, Mx: sample mass, V: sample volume, 500. mu.L: internal standard concentration 20ng/mL), and each substance is expressed as a relative amount (ng/g).
3.3 results of the experiment
When external pathogenic factors invade, the body can release different types of inflammation mediators, the steady state of endogenous metabolites in the body is influenced, the balance of proinflammatory and anti-inflammatory phospholipid mediators is imbalanced, inflammation is caused, the content change of the endogenous metabolites of Arachidonic acid can well reflect the change trend of the body inflammation, the influence of toad tryptamine quaternary ammonium salt on foot inflammation mediators of formalin pain-induced model mice is shown in Table 11, experimental results show that compared with blank groups, the right foot inflammation mediators of model groups of mice are obviously improved, and include dhkPGF2 α, 20-ethyl PGF2a, 1a,1b-dihomo-PGE1, dhk PGE2, 8-HETrE, 11,12-EET, 8,9-EET, 13-HpODE, 13-HDoHE, Arachidonic acid and the like, after the toad tryptamine quaternary ammonium salt is given, the inflammation mediators are obviously reduced, the blank groups (486.04 + -11.64 ng/g) 20-HDoHE, Arachidonic acid, and the like, and the administration of toad tryptamine quaternary ammonium salt is reduced in the blank groups (486.04 + -11, 24, 23, 11, 23, 5, 3, 5 and 5, 3, 5, 3, 5, 3.
TABLE 11 relative amounts of inflammatory substances in Right paw of mice given toad tryptamine Quaternary ammonium salt: (
n＝6，ng/g)
Note: p is less than or equal to 0.01, and has extremely significant difference compared with the model group; p is less than or equal to 0.05, and there is a significant difference compared with the model group.
##P is less than or equal to 0.01, and has very obvious difference compared with the normal saline group;
#p is less than or equal to 0.05, and the difference is obvious compared with the normal saline group. (for model group only)
Example 6 comparison of Central analgesic Effect of bufotenine, bufotenine Quaternary ammonium salt, and serotonin
1.1 materials and methods
1.2 results of the experiment
The result shows that compared with the group before administration, the pain threshold value of the bufotamine quaternary ammonium salt (5mg/kg) after administration is obviously improved (P is less than 0.05) 1 hour after administration, and the compound preparation shows a certain analgesic effect; both bufotenine and serotonin have no analgesic effect at the same test doses as described below.
TABLE 12 comparison of Central analgesic effects of bufotamine, bufotamine Quaternary ammonium salt, and serotonin ((
n＝10-12)
Note: p is less than or equal to 0.05, and is obviously different from the group before administration.
Example 7 synergistic analgesic Effect of combination of bufotenine Quaternary ammonium salt and morphine
1.1 materials and methods
1.2 Experimental results and discussion
Effect of combination of bufotenine and morphine on hind paw thermalgia threshold of mice
Three groups of drugs failed to prolong the pain threshold of the hind paw of the mice to the hot plate when administered intraperitoneally for half an hour at doses of morphine hydrochloride (0.075mg/kg) and bufotenine quaternary ammonium salt (5,10 mg/kg). The pain threshold extension values of mice in the toad venom tryptamine quaternary ammonium salt (5,10mg/kg) group are only 2.9 percent and 13.5 percent, which shows that the pain relieving effect cannot be generated 30min after the following medicaments are used alone under the selected dosage. However, when the drugs with the doses are combined (morphine + toad poison tryptamine quaternary ammonium salt), compared with the drugs before administration, the pain threshold elongation value of the mice is remarkably increased to 30.4 percent and 48.6 percent (P is less than 0.05), and the combined group has remarkable analgesic effect. The result shows that the combination of the toad venom tryptamine quaternary ammonium salt and the morphine has the synergistic effect.
TABLE 13 Central analgesic Effect of combination of bufotenine and morphine: (
n＝10-12)
The above embodiments are merely preferred embodiments of the present invention, which are provided for illustrating the technical concept and features of the present invention, and the purpose thereof is to enable those skilled in the art to understand the present invention and implement the present invention, and not to limit the scope of the present invention, and any modification, equivalent replacement, improvement, etc. made in accordance with the spirit of the present invention should be included in the scope of the present invention.
Claims (10)
1. A synthetic method of bufotenine is characterized by comprising the following steps:
a. in an organic solvent, 5-benzyloxy indole (S-0) is used as a starting material, and 2- (5- (benzyloxy) -1H-indol-3-yl) -2-oxo-acetyl halide (S-1) is generated under the acylation action of oxalyl halide, wherein X is halogen;
b. mixing the 2- (5- (benzyloxy) -1H-indol-3-yl) -2-oxoacetoacetyl halide produced above with dimethylamine or a salt thereof in a solvent in the presence of a base to react and obtain 2- (5- (benzyloxy) -1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide (S-2);
c. catalytically hydrogenating the 2- (5- (benzyloxy) -1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide produced above in an organic solvent in the presence of a hydrogenation catalyst to remove benzyl groups to give 2- (5-hydroxy-1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide (S-3);
d. reducing the ortho carbonyl of the generated 2- (5-hydroxy-1H-indol-3-yl) -N, N-dimethyl-2-oxoacetamide into methylene under the action of a reducing agent to obtain the bufotenine (S-4).
2. The method for synthesizing bufotenine according to claim 1, wherein in step a, X in S-1 is chlorine or bromine; the oxalyl halide is selected from oxalyl chloride and oxalyl bromide, preferably oxalyl chloride, wherein the molar ratio of S-0 to oxalyl halide is 1: 1-1: 5, preferably 1: 1-1: 3; the organic solvent is anhydrous organic solvent selected from C
1-6Alcohol, C
1-6Ethers or cyclic ethers, C
1-6Esters, C
1-6Ketones, C
1-6Halogenated alkanes, C
1-6One or more of alkyl nitrile and DMF, preferably one or more of ethyl acetate, dichloromethane, methanol, diethyl ether, tetrahydrofuran, DMF, ethanol, acetonitrile and acetone solution, more preferably ethyl acetate, diethyl ether and tetrahydrofuran; the reaction time is 0.25-3 h, preferably 0.5-1 h; the temperature is 0-80 deg.C, preferably 10-50 deg.C, more preferably 20-40 deg.C; preferably, X in S-1 is chlorine; the oxalyl halide is oxalyl chloride, wherein the molar ratio of S-0 to oxalyl halide is 1: 1-1: 3; the organic solvent is ethyl acetate, diethyl ether and tetrahydrofuran; the reaction time is 0.5-1 h.
3. The method for synthesizing bufotenine according to claim 1, wherein in step b, the base is selected from alkali metal hydroxide, alkaline earth metal hydroxide, carbonate of alkali metal or alkaline earth metal, and C of alkali metal or alkaline earth metal
1-6Alkoxide, triethylamine, diisopropylethylamine, pyridine and N, N-dimethylaminopyridine or a plurality of alkoxides; preferably one or more of sodium hydroxide, potassium hydroxide, magnesium hydroxide, calcium hydroxide, sodium carbonate, potassium carbonate, magnesium carbonate, calcium carbonate, sodium methoxide, sodium ethoxide, magnesium methoxide, triethylamine, diisopropylethylamine, pyridine and N, N-dimethylaminopyridine; more preferably sodium hydroxide, potassium hydroxide, magnesium hydroxide, calcium hydroxide, sodium carbonate, potassium carbonate, magnesium carbonate, calcium carbonate, sodium methoxide, sodium ethoxide and mixtures thereof; sodium hydroxide, potassium hydroxide, calcium hydroxide and mixtures thereof are particularly preferred; wherein the molar ratio of S-1 to the alkali is 1: 1-15, preferably 1:1-10, and more preferably 1: 1-8; the molar ratio of S-1 to dimethylamine or salts thereof is 1: 1-8, preferably 1: 1-5, wherein the dimethylamine salt is selected from inorganic salts of dimethylamine, preferably hydrochloride, bromate, sulfate and phosphate; the solvent is selected from polar solvents, preferably water, C
1-6Alcohol, C
1-6Ketones, C
1-6Ether, DMF, DMSO; more preferably water, C
1-6An alcohol; water, methanol, ethanol are particularly preferred; most preferably water; the reaction time is 0.2-3 h, preferably 0.5-1 h; the temperature is 0-60 deg.C, preferably 10-50 deg.C, more preferably 20-40 deg.C; preferably, the base is selected from the group consisting of sodium hydroxide, potassium hydroxide, sodium hydroxide,calcium hydroxide, wherein the molar ratio of S-1 to alkali is 1: 1-8; the molar ratio of S-1 to dimethylamine or salts thereof is 1: 1-5, wherein dimethylamine salt is selected from dimethylamine hydrochloride; the solvent is selected from water; the reaction time is 0.5-1 h.
4. The method for synthesizing bufotoxin tryptamine according to claim 1, wherein in step c, the hydrogenation catalyst is selected from palladium catalyst and platinum catalyst, preferably palladium catalyst in supported form; more preferably Pd/C, Pd (OH)
2/C、PdO/C、PdO/Al
2O
3、PdO/SiO
2、Pd/CaCO
3(ii) a Pd/C, Pd (OH) is particularly preferred
2/C, PdO/C; most preferably Pd/C; wherein the loading of palladium is in the range of 0.5% to 25%, preferably 0.5% to 25%, more preferably 1% to 20%, most preferably 5 to 15%; the mass ratio of the catalyst to the S-2 is 1: 4-50, preferably 1: 5-20, and more preferably 1: 5-10; the catalytic hydrogenation is carried out in the presence of hydrogen, the pressure of the hydrogen being from normal pressure to 10 MPa; the temperature of the catalytic hydrogenation reaction is 0 to 80 ℃, preferably 10 to 50 ℃, more preferably 20 to 40 ℃; the organic solvent is selected from C
1-6Alcohol, C
1-6Ethers or cyclic ethers, C
1-6Esters, C
1-6Ketones, C
1-6Halogenated alkanes, C
1-6One or more of alkyl nitrile and DMF, preferably one or more of ethyl acetate, dichloromethane, methanol, diethyl ether, tetrahydrofuran, DMF, ethanol, acetonitrile and acetone solution; more preferably two or more of ethyl acetate, dichloromethane, methanol, diethyl ether, tetrahydrofuran, DMF, ethanol, acetonitrile and acetone solution; particularly preferably two of ethyl acetate, dichloromethane, methanol, diethyl ether, tetrahydrofuran, DMF, ethanol, acetonitrile and acetone solution, and most preferably a mixed solvent of methanol and tetrahydrofuran, wherein the ratio of the two solvents is 0.1:1-10, preferably 0.5:1-5, more preferably 0.5:1-2, most preferably 0.5:1, 1:1,1: 2; preferably, the hydrogenation catalyst is selected from Pd/C; wherein the loading of palladium is in the range of 5 to 15%; the mass ratio of the catalyst to the S-2 is 1: 5-10; the temperature is 20-40 ℃; the organic solvent is selected from mixed solvent of methanol and tetrahydrofuran, wherein the ratio of the two solvents is 0.5: 1-2.
5. The method for synthesizing bufotenine according to claim 1, wherein in step d, the reducing agent is selected from sodium borohydride, lithium aluminum hydride, diisobutylaluminum hydride, lithium tri-sec-butylborohydride, sodium bis (2-methoxyethoxy) aluminum hydride, sodium cyanoborohydride, and trialkoxyaluminum hydride; preferably sodium borohydride, lithium aluminum hydride, diisobutylaluminum hydride, lithium tri-sec-butylborohydride; more preferably sodium borohydride, lithium aluminum hydride; the molar ratio of reducing agent to S-3 is 1-25:1, preferably 5-20:1, more preferably 6-15: 1; the solvent is selected from C
1-6Alcohol, C
1-6Ethers or cyclic ethers, C
1-6Esters, C
1-6One or more of alkyl nitrile or toluene; preferably one or more of methanol, ethanol, ethyl acetate, methyl tert-butyl ether, diethyl ether or tetrahydrofuran, more preferably methanol, ethanol or tetrahydrofuran; the reaction temperature is from-20 ℃ to 80 ℃, preferably from-10 ℃ to 50 ℃, more preferably from 0 ℃ to 40 ℃; step d is optionally carried out under nitrogen or argon atmosphere; preferably, the reducing agent is selected from sodium borohydride, lithium aluminum hydride; the molar ratio of the reducing agent to the S-3 is 6-15: 1; the solvent is selected from methanol, ethanol or tetrahydrofuran; the reaction temperature is 0 ℃ to 40 ℃.
6. A synthetic method of a bufotenine quaternary ammonium salt is characterized by comprising the following steps:
preparing bufotenine according to the process of claim 1;
e. in an organic solvent, reacting bufotoxin tryptamine with a methylation reagent to obtain 2- (5-hydroxy-1H-indol-3-yl) -N, N, N-trimethylethan-1-amine chloride.
7. The method for synthesizing the bufotenine quaternary ammonium salt according to claim 6, wherein in the step e, the methylating agent is selected from methyl iodide, methyl bromide, methyl chloride, methyl trifluoromethanesulfonate and dimethyl sulfate; preferably methyl iodide, methyl bromide; more preferably iodomethaneAn alkane; the molar ratio of methylating agent to S-4 is from 1 to 30:1, preferably from 5 to 25:1, more preferably from 10 to 15: 1; the organic solvent is selected from C
1-6Alcohol, C
1-6Esters, C
1-6Ketones, C
1-6Ethers or cyclic ethers, C
1-6One or more alkylnitriles, DMF, or DMSO, preferably one or more of methanol, ethanol, ethyl acetate, acetone, methyl tert-butyl ether, diethyl ether, acetonitrile, or tetrahydrofuran, more preferably methanol, ethanol; the reaction temperature is 0-60 ℃, preferably 10-50 ℃, and more preferably 20-40 ℃; step e is optionally carried out under dark conditions; optionally adding silver chloride after the reaction is completed; preferably, the methylating agent is selected from methyl iodide; the molar ratio of the methylating agent to the S-4 is 10-15: 1; the organic solvent is selected from methanol and ethanol; the reaction temperature is 20-40 ℃; step e is carried out under the condition of keeping out of the sun; and silver chloride is added after the reaction is complete.
8. The bufotoxin tryptamine or the bufotoxin tryptamine quaternary ammonium salt is applied to the preparation of analgesic or anti-inflammatory drugs, and preferably, the bufotoxin tryptamine quaternary ammonium salt is 2- (5-hydroxy-1H-indol-3-yl) -N, N, N-trimethylethan-1-amine chloride.
9. A pharmaceutical composition comprises bufotenine or bufotenine quaternary ammonium salt and opioid such as morphine hydrochloride, preferably, the bufotenine quaternary ammonium salt is 2- (5-hydroxy-1H-indol-3-yl) -N, N, N-trimethylethan-1-amine chloride.
10. The application of the bufotenine or the bufotenine quaternary ammonium salt in preparing the analgesic drug is characterized in that the bufotenine or the bufotenine quaternary ammonium salt and pharmaceutically acceptable auxiliary materials are prepared into a pharmaceutical preparation, preferably, the bufotenine quaternary ammonium salt is 2- (5-hydroxy-1H-indol-3-yl) -N, N, N-trimethylethan-1-amine chloride.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911259232.8A CN110776454B (en) | 2019-12-10 | 2019-12-10 | Synthesis method of bufotenine and quaternary ammonium salt thereof and application of bufotenine and quaternary ammonium salt thereof in preparation of analgesic and anti-inflammatory drugs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911259232.8A CN110776454B (en) | 2019-12-10 | 2019-12-10 | Synthesis method of bufotenine and quaternary ammonium salt thereof and application of bufotenine and quaternary ammonium salt thereof in preparation of analgesic and anti-inflammatory drugs |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110776454A true CN110776454A (en) | 2020-02-11 |
| CN110776454B CN110776454B (en) | 2021-03-23 |
Family
ID=69394326
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911259232.8A Active CN110776454B (en) | 2019-12-10 | 2019-12-10 | Synthesis method of bufotenine and quaternary ammonium salt thereof and application of bufotenine and quaternary ammonium salt thereof in preparation of analgesic and anti-inflammatory drugs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110776454B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114432247A (en) * | 2022-01-18 | 2022-05-06 | 南京中医药大学 | Toad tryptamine liposome and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001054681A2 (en) * | 2000-01-27 | 2001-08-02 | Massachusetts Institute Of Technology | Composition for treatment of stress |
| WO2002036114A1 (en) * | 2000-11-01 | 2002-05-10 | Respiratorius Ab | Composition comprising serotonin receptor antagonists, 5 ht-2 and 5 ht-3 |
| US20080293695A1 (en) * | 2007-05-22 | 2008-11-27 | David William Bristol | Salts of physiologically active and psychoactive alkaloids and amines simultaneously exhibiting bioavailability and abuse resistance |
-
2019
- 2019-12-10 CN CN201911259232.8A patent/CN110776454B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001054681A2 (en) * | 2000-01-27 | 2001-08-02 | Massachusetts Institute Of Technology | Composition for treatment of stress |
| WO2002036114A1 (en) * | 2000-11-01 | 2002-05-10 | Respiratorius Ab | Composition comprising serotonin receptor antagonists, 5 ht-2 and 5 ht-3 |
| US20080293695A1 (en) * | 2007-05-22 | 2008-11-27 | David William Bristol | Salts of physiologically active and psychoactive alkaloids and amines simultaneously exhibiting bioavailability and abuse resistance |
Non-Patent Citations (5)
| Title |
|---|
| JAMES L. COLLINS III，等: "Calixarene-mediated liquid membrane transport of choline conjugates3: The effect of handle variation on neurotransmitter transport", 《BIOORGANIC & MEDICINAL CHEMISTRY LETTERS》 * |
| L.F. DE OLIVEIRA，等: "BLOCKADE BY BUFOTENIDINE OF THE SEROTONIN- AND NARCOTIC-ANALGESICS-INDUCED CONTRACTION OF DOG INTESTINE IN VIVO", 《EUROPEAN JOURNAL OF PHARMACOLOGY》 * |
| WATKINS, L. R.等: "Reversal of spinal cord non-opiate analgesia by conditioned anti-analgesia in the rat", 《PAIN》 * |
| YU ZHANG，等: "The protection of indolealkylamines from LPS-induced inflammation in zebrafish", 《JOURNAL OF ETHNOPHARMACOLOGY》 * |
| YU-YUN WANG，等: "Synthesis of a deuterium-labelled standard of bufotenine(5-HO-DMT)", 《JOURNAL OF LABELLED COMPOUNDS AND RADIOPHARMACEUTICALS》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114432247A (en) * | 2022-01-18 | 2022-05-06 | 南京中医药大学 | Toad tryptamine liposome and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110776454B (en) | 2021-03-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP3341007B1 (en) | Malt1 inhibitors and uses thereof | |
| US10441553B2 (en) | Fluorinated CBD compounds, compositions and uses thereof | |
| KR20190038616A (en) | TLR7 / 8 antagonists and their uses | |
| SA518391218B1 (en) | Isoindolinone inhibitors of the MDM2-P53 interaction having anticancer activity | |
| CN1882581A (en) | Derivatives of n-[heteroaryl(piperidine-2-yl)methyl]benzamide, preparation method thereof and application of same in therapeutics | |
| CN1043763C (en) | Tropane-z-aldoxine derivatives and preparation process and use thereof | |
| CN1181077A (en) | Benzopyran-containing compounds and method for their use | |
| JP2009520697A (en) | Compounds having cytokine regulatory properties | |
| CN102459177B (en) | The dibasic Arylsulfonamide ccr 3 antagonists of 2,5- | |
| JP3529736B2 (en) | Hexahydropyrazolo [4,3-c] pyridine metabolite | |
| CN110776454B (en) | Synthesis method of bufotenine and quaternary ammonium salt thereof and application of bufotenine and quaternary ammonium salt thereof in preparation of analgesic and anti-inflammatory drugs | |
| CN1907291A (en) | Injectio for nanometer notoginsenoside and its preparing method | |
| WO2021255174A1 (en) | Small molecule modulators of il-17 | |
| CN114746095A (en) | HCK as therapeutic target of MYD88 mutant disease | |
| CN106256830B (en) | A kind of deuterated IDO inhibitor and its preparation method and application | |
| JP2016508489A (en) | Protopanaxadiol derivative, production method thereof and application thereof | |
| CN1902671A (en) | Language phonetic system | |
| CN1763030A (en) | Puerarin derivate and medical usage thereof | |
| EP3279200A1 (en) | Crystal of 5-((2-(6-amino)-9h-purin-9-yl)ethyl)amino)pentan-1-ol | |
| CN1876000A (en) | 'Yan Lu Ru Kang' pharmaceutical preparation for treating mammary gland hyperplasia, its preparation process and quality control method | |
| JP2023514291A (en) | Crystal forms of JAK inhibitors and uses thereof | |
| EP1793834B1 (en) | Indolin-2-one pyridine derivatives, preparation and therapeutic use thereof | |
| CN1857385A (en) | Medicine composition for treating cervical spondylosis and its preparing method | |
| CN1720248A (en) | Crystalline fumarate salts of 1-azabicyclo[2.2.2]oct substituted furo[2,3-c]pyridinyl carboxamide and compositions and preparations thereof | |
| CN111233877B (en) | Galanthamine pamoate and preparation method thereof |
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 |