CN114907228B - Colchicine and magnolol compound, synthesis method thereof and application thereof in resisting new coronaviruses - Google Patents
Colchicine and magnolol compound, synthesis method thereof and application thereof in resisting new coronaviruses Download PDFInfo
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- CN114907228B CN114907228B CN202210732889.7A CN202210732889A CN114907228B CN 114907228 B CN114907228 B CN 114907228B CN 202210732889 A CN202210732889 A CN 202210732889A CN 114907228 B CN114907228 B CN 114907228B
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- IAKHMKGGTNLKSZ-INIZCTEOSA-N (S)-colchicine Chemical compound C1([C@@H](NC(C)=O)CC2)=CC(=O)C(OC)=CC=C1C1=C2C=C(OC)C(OC)=C1OC IAKHMKGGTNLKSZ-INIZCTEOSA-N 0.000 title claims abstract description 134
- 229960001338 colchicine Drugs 0.000 title claims abstract description 67
- VVOAZFWZEDHOOU-UHFFFAOYSA-N honokiol Natural products OC1=CC=C(CC=C)C=C1C1=CC(CC=C)=CC=C1O VVOAZFWZEDHOOU-UHFFFAOYSA-N 0.000 title claims abstract description 62
- -1 magnolol compound Chemical class 0.000 title claims abstract description 39
- 241000711573 Coronaviridae Species 0.000 title claims abstract description 10
- 238000001308 synthesis method Methods 0.000 title abstract description 5
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims abstract description 62
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 48
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000003513 alkali Substances 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 15
- 238000002156 mixing Methods 0.000 claims abstract description 13
- 150000001413 amino acids Chemical class 0.000 claims abstract description 11
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims abstract description 9
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229940079593 drug Drugs 0.000 claims abstract description 5
- 239000003814 drug Substances 0.000 claims abstract description 5
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 5
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 54
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 33
- 239000000243 solution Substances 0.000 claims description 22
- 238000006243 chemical reaction Methods 0.000 claims description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 239000007787 solid Substances 0.000 claims description 14
- 238000005406 washing Methods 0.000 claims description 13
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 239000012071 phase Substances 0.000 claims description 9
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 8
- 239000000126 substance Substances 0.000 claims description 7
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 238000010898 silica gel chromatography Methods 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 5
- 239000012074 organic phase Substances 0.000 claims description 5
- 238000000746 purification Methods 0.000 claims description 4
- 238000002390 rotary evaporation Methods 0.000 claims description 4
- 239000003599 detergent Substances 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 238000010791 quenching Methods 0.000 claims description 3
- 238000012986 modification Methods 0.000 abstract description 4
- 230000004048 modification Effects 0.000 abstract description 4
- 230000000694 effects Effects 0.000 description 12
- 239000000047 product Substances 0.000 description 7
- 101800000535 3C-like proteinase Proteins 0.000 description 6
- 101800002396 3C-like proteinase nsp5 Proteins 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 238000001514 detection method Methods 0.000 description 4
- 108010028275 Leukocyte Elastase Proteins 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 208000028399 Critical Illness Diseases 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 102000016799 Leukocyte elastase Human genes 0.000 description 2
- 241000234280 Liliaceae Species 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 239000008346 aqueous phase Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- BQJCRHHNABKAKU-KBQPJGBKSA-N morphine Chemical compound O([C@H]1[C@H](C=C[C@H]23)O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4O BQJCRHHNABKAKU-KBQPJGBKSA-N 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- 206010000748 Acute febrile neutrophilic dermatosis Diseases 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 208000009137 Behcet syndrome Diseases 0.000 description 1
- 208000008439 Biliary Liver Cirrhosis Diseases 0.000 description 1
- 208000033222 Biliary cirrhosis primary Diseases 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 241000206576 Chondrus Species 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010016207 Familial Mediterranean fever Diseases 0.000 description 1
- 241000131505 Gloriosa Species 0.000 description 1
- 201000005569 Gout Diseases 0.000 description 1
- 101000851058 Homo sapiens Neutrophil elastase Proteins 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical group Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 206010021118 Hypotonia Diseases 0.000 description 1
- 244000056623 Iphigenia indica Species 0.000 description 1
- 241001673966 Magnolia officinalis Species 0.000 description 1
- 208000037581 Persistent Infection Diseases 0.000 description 1
- 208000012641 Pigmentation disease Diseases 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 206010065159 Polychondritis Diseases 0.000 description 1
- 208000012654 Primary biliary cholangitis Diseases 0.000 description 1
- 201000004681 Psoriasis Diseases 0.000 description 1
- 241000219061 Rheum Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 208000010265 Sweet syndrome Diseases 0.000 description 1
- 201000009594 Systemic Scleroderma Diseases 0.000 description 1
- 206010042953 Systemic sclerosis Diseases 0.000 description 1
- 206010047115 Vasculitis Diseases 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 206010002022 amyloidosis Diseases 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000767 anti-ulcer Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 206010015037 epilepsy Diseases 0.000 description 1
- 102000052502 human ELANE Human genes 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 229960005181 morphine Drugs 0.000 description 1
- 230000036640 muscle relaxation Effects 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 238000012805 post-processing Methods 0.000 description 1
- CHKVPAROMQMJNQ-UHFFFAOYSA-M potassium bisulfate Chemical compound [K+].OS([O-])(=O)=O CHKVPAROMQMJNQ-UHFFFAOYSA-M 0.000 description 1
- 229910000343 potassium bisulfate Inorganic materials 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 208000022256 primary systemic amyloidosis Diseases 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000012265 solid product Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C233/00—Carboxylic acid amides
- C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
- C07C233/34—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by amino groups
- C07C233/41—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by amino groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a ring other than a six-membered aromatic ring
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
- A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
- A61K31/222—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin with compounds having aromatic groups, e.g. dipivefrine, ibopamine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
- A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
- A61K31/223—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin of alpha-aminoacids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C231/00—Preparation of carboxylic acid amides
- C07C231/12—Preparation of carboxylic acid amides by reactions not involving the formation of carboxamide groups
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2603/00—Systems containing at least three condensed rings
- C07C2603/02—Ortho- or ortho- and peri-condensed systems
- C07C2603/04—Ortho- or ortho- and peri-condensed systems containing three rings
- C07C2603/30—Ortho- or ortho- and peri-condensed systems containing three rings containing seven-membered rings
- C07C2603/34—Benzoheptalenes; Hydrogenated benzoheptalenes
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
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Abstract
The invention belongs to the technical field of colchicine modification, and provides a colchicine and magnolol compound, a synthesis method thereof and application thereof in resisting new coronaviruses. The method provided by the invention comprises the following steps: (1) Mixing colchicine, amino acid, alkali solution and ethanol, and reacting to generate amino acid modified colchicine; (2) Mixing and reacting amino acid modified colchicine, magnolol, methylene dichloride, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and 4-dimethylaminopyridine to generate a colchicine and magnolol compound. The invention adopts magnolol to modify colchicine for the first time. The chemical synthesis method of the colchicine and magnolol compound provided by the invention is simple to operate and high in yield. The obtained colchicine and magnolol compound can be used for preparing anti-novel coronavirus medicines.
Description
Technical Field
The invention relates to the technical field of colchicine modification, in particular to a colchicine and magnolol compound, a synthesis method thereof and application thereof in resisting new coronaviruses.
Background
Colchicine is an alkaloid extracted from corms and seeds of colchicine, a plant of the family Liliaceae, and also present in the plants of the family Liliaceae, gloriosa and Iphigenia indica. The structural formula is shown as formula A:
colchicine has good clinical curative effect and plays a vital role in treating acute gout, behcet's disease, familial mediterranean fever, chondrus calcareous pigmentation disease, systemic scleroderma, amyloidosis and the like; and has application value in the treatment of cardiovascular diseases, dermatitis, primary biliary cirrhosis, sweet's syndrome, psoriasis, chronic infection, amyloidosis, recurrent polychondritis, alzheimer's disease, epilepsy, allergic diseases, necrotic and leucocyte destructive vasculitis and other diseases (J.Med. Chem.2020,63,10618). Notably, colchicine is effective in reducing mortality in critically ill patients (ann.rheum.dis., 2020,79,1286) in the treatment of critically ill patients with new coronaries, showing great potential for application.
However, when in clinical medication, colchicine is easy to bring severe side effects such as diarrhea and vomiting, and in the aspect of anti-tumor research, colchicine is difficult to directly apply because of the great toxicity to normal cells, so that it is important to carry out proper structural modification on colchicine so as to weaken or even eliminate the side effects.
Magnolol (magnolol) is separated from Magnolia officinalis, has obvious and durable central muscle relaxation, central nerve inhibition, antiinflammatory, antibacterial, pathogenic microorganism resisting, antiulcer, antioxidant, antitumor, morphine withdrawal inhibiting, and platelet aggregation inhibiting effects. The structural formula is shown as formula B:
at present, the technology of splicing colchicine and magnolol has not been reported yet.
Disclosure of Invention
The invention aims to provide a colchicine and magnolol compound, a synthesis method thereof and application thereof in resisting new coronaviruses so as to make up for the blank of the prior art.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a chemical synthesis method of colchicine and magnolol compound, which comprises the following steps:
(1) Mixing colchicine, amino acid, alkali solution and ethanol, and reacting to generate amino acid modified colchicine;
(2) Mixing and reacting amino acid modified colchicine, magnolol, methylene dichloride, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and 4-dimethylaminopyridine to generate a colchicine and magnolol compound.
Preferably, the amino acid has the structural formulaWherein n is greater than or equal to 1.
Preferably, the molar ratio of colchicine to amino acid is 5 (8-12);
the molar volume ratio of colchicine to ethanol is 5mmol: (8-13) mL;
the mass volume ratio of alkali and water in the alkali solution is (0.3-0.6) g:10mL of the alkali solution is sodium hydroxide aqueous solution or potassium hydroxide aqueous solution;
the volume ratio of the alkali solution to the ethanol is 10: (8-13).
Preferably, the temperature of the reaction in the step (1) is 60-70 ℃ and the time is 3-7 hours.
Preferably, the molar ratio of colchicine to magnolol is 5 (8-12);
the molar ratio of colchicine to 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride is 5 (8-12);
the molar ratio of colchicine to 4-dimethylaminopyridine is 5 (0.3-0.8);
the molar volume ratio of colchicine to dichloromethane is 5mmol: (40-60) mL.
Preferably, the temperature of the reaction in the step (2) is room temperature and the time is 10 to 15 hours.
Preferably, after the reaction in the step (1) is finished, purifying the obtained product system to obtain pure amino acid modified colchicine for the step (2);
the purification treatment comprises the following steps:
performing rotary evaporation on the obtained product system to remove ethanol and obtain a solid substance;
washing the obtained solid substance with ethyl acetate, retaining a water phase, adding acid to adjust the pH value to 4-5, and separating out white solid;
washing the obtained white solid with dichloromethane for three times, drying with anhydrous sodium sulfate, and concentrating to obtain pure amino acid modified colchicine;
the volume ratio of the ethyl acetate to the ethanol in the step (1) is 15: (8-13);
the volume ratio of dichloromethane used for each washing and ethanol used in step (1) was independently 30: (8-13).
Preferably, after the reaction in the step (2) is finished, carrying out post-treatment on the product system to obtain a pure colchicine and magnolol compound;
the post-treatment comprises the following steps:
adding water to quench the reaction, and washing and separating the solution to obtain a water phase;
extracting the obtained water phase with dichloromethane for three times, mixing the organic phases, drying with anhydrous sodium sulfate, concentrating, and subjecting to silica gel column chromatography to obtain pure colchicine and magnolol compound;
the volume ratio of dichloromethane used for each extraction to ethanol used in step (1) was independently 50: (8-13);
the detergent used in the silica gel column chromatography is a mixture of ethyl acetate and methanol, and the volume ratio of the ethyl acetate to the methanol is (8-12): 1.
The invention also provides the colchicine and magnolol compound obtained by the method.
The invention also provides application of the colchicine and magnolol compound in preparation of anti-new coronavirus medicines.
The invention adopts magnolol to modify colchicine for the first time. The chemical synthesis method of the colchicine and magnolol compound provided by the invention is simple to operate and high in yield. The obtained colchicine and magnolol compound can be used for preparing anti-novel coronavirus medicines.
Drawings
FIG. 1 is a graph showing that colchicine and magnolol complexes significantly inhibit the activity of the main protease;
FIG. 2 is a graph showing that colchicine and magnolol complexes significantly inhibit neutrophil elastase protease activity.
Detailed Description
The invention provides a chemical synthesis method of colchicine and magnolol compound, which comprises the following steps:
(1) Mixing colchicine, amino acid, alkali solution and ethanol, and reacting to generate amino acid modified colchicine;
(2) Mixing and reacting amino acid modified colchicine, magnolol, methylene dichloride, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and 4-dimethylaminopyridine to generate a colchicine and magnolol compound.
In the present invention, the amino acid has the structural formulaWherein n.gtoreq.1, is preferably 1 to 20, more preferably 1 to 10, still more preferably 1 to 4, and may specifically be 1, 2, 3 or 4.
In the invention, the molar ratio of colchicine to amino acid is 5 (8-12), preferably 5 (9-11), and more preferably 5:10; the molar volume ratio of colchicine to ethanol is 5mmol: (8-13) mL, preferably 5mmol: (9-12) mL, more preferably 5mmol: (10-11) mL; the mass volume ratio of alkali and water in the alkali solution is (0.3-0.6) g:10mL, preferably (0.4 to 0.5) g:10mL; the volume ratio of the alkali solution to the ethanol is 10: (8-13), preferably 10: (9 to 12), more preferably 10: (10-11).
In the present invention, the temperature of the reaction in the step (1) is 60 to 70 ℃, preferably 64 to 68 ℃, and more preferably 65 to 66 ℃; the time is 3 to 7 hours, preferably 4 to 6 hours, and more preferably 5 hours.
In the invention, the molar ratio of colchicine to magnolol is 5 (8-12), preferably 5 (9-11), and more preferably 5:10; the molar ratio of colchicine to 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride is 5 (8-12), preferably 5 (9-11), more preferably 5:10; the molar ratio of colchicine to 4-dimethylaminopyridine is 5 (0.3-0.8), preferably 5 (0.4-0.7), and more preferably 5 (0.5-0.6); the molar volume ratio of colchicine to dichloromethane is 5mmol: (40-60) mL, preferably 5mmol: (45-55) mL, more preferably 5mmol: (50-53) mL.
In the present invention, the temperature of the reaction in the step (2) is room temperature for 10 to 15 hours, preferably 11 to 14 hours, and more preferably 12 to 13 hours.
In the invention, after the reaction of the step (1), purifying the obtained product system to obtain pure amino acid modified colchicine for the step (2);
the purification treatment comprises the following steps:
performing rotary evaporation on the obtained product system to remove ethanol and obtain a solid substance;
washing the obtained solid substance with ethyl acetate, retaining a water phase, adding acid to adjust the pH value to 4-5, and separating out white solid; the acid used for regulating the pH value is dilute hydrochloric acid, dilute sulfuric acid or potassium hydrogen sulfate aqueous solution;
washing the obtained white solid with dichloromethane for three times, drying with anhydrous sodium sulfate, and concentrating to obtain pure amino acid modified colchicine;
the volume ratio of the ethyl acetate to the ethanol in the step (1) is 15: (8-13), preferably 15: (10-11);
the volume ratio of dichloromethane used for each washing and ethanol used in step (1) was independently 30: (8-13), preferably 30: (10-12).
In the invention, after the reaction of the step (2) is finished, post-processing is carried out on a product system to obtain a pure colchicine and magnolol compound;
the post-treatment comprises the following steps:
adding water to quench the reaction, and washing and separating the solution to obtain a water phase; specifically, adding water equivalent to the system after finishing, and washing the reaction solution; re-separating the liquid and retaining the organic phase;
extracting the obtained water phase with dichloromethane for three times, mixing the organic phases, drying with anhydrous sodium sulfate, concentrating, and subjecting to silica gel column chromatography to obtain pure colchicine and magnolol compound;
the volume ratio of dichloromethane used for each extraction to ethanol used in step (1) was independently 50: (8-13), preferably 50: (10-12);
the detergent used in the silica gel column chromatography is a mixture of ethyl acetate and methanol, and the volume ratio of the ethyl acetate to the methanol is (8-12): 1, preferably (9-10): 1.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
The reactions for the various embodiments of the present invention are as follows:
wherein D1 is example 1, D2 is example 2, D3 is example 3, and D4 is example 4.
Colchicine (2.0 g,5 mmol) and amino acids of different chain lengths (10 mmol) were added to ethanol (10 mL), and then sodium hydroxide (0.4 g, dissolved in 10mL of water) was added and reacted at 70℃for 5 hours. The ethanol solvent was removed by rotary evaporation, washed once with ethyl acetate (15 mL), the aqueous phase was retained, the pH was adjusted to 4.5 with acid, white solid precipitated, and washed three times with dichloromethane (30 mL each). Dried over anhydrous sodium sulfate and concentrated. The resulting solid product C was dissolved in methylene chloride (50 mL), magnolol (2.6 g,10 mmol), 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI, 1.9g,10 mmol) and 4-dimethylaminopyridine (DMAP, 61mg,0.5 mmol) were added. The reaction was continued at room temperature for 12 hours, quenched with water, the separated liquid was washed, and the aqueous phase was extracted three times with dichloromethane (50 mL each). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated and chromatographed on a silica gel column (ethyl acetate/methanol, volume ratio 10:1) to give product D as a tan solid.
D1:2.9g, yield 84%; r is R f =0.4 (ethyl acetate/methanol, volume ratio 10:1); 1 H NMR (400MHz,DMSO-d 6 )δ9.36(s,1H),8.57(d,J＝7.6Hz,1H),7.77(t,J＝6.3Hz, 1H),7.25–7.10(m,4H),7.06(d,J＝11.0Hz,1H),6.96(dd,J＝8.3,2.3Hz,1H),6.90–6.81(m,2H),6.76(s,1H),6.12(d,J＝11.2Hz,1H),5.98(ddt,J＝16.9, 10.0,6.8Hz,1H),5.85(ddt,J＝16.8,9.9,6.8Hz,1H),5.13(dq,J＝17.1,1.7Hz,1H),5.06(ddd,J＝10.0,2.2,1.2Hz,1H),4.98(dt,J＝17.0,1.8Hz,1H),4.93 (ddt,J＝10.0,2.4,1.4Hz,1H),4.37(dt,J＝12.1,7.2Hz,1H),4.30(dd,J＝18.5,6.2Hz,1H),4.18(dd,J＝18.4,6.3Hz,1H),3.84(s,3H),3.82(s,3H),3.48(s, 3H),3.41(d,J＝6.9Hz,2H),3.21(d,J＝6.8Hz,2H),2.55(dd,J＝12.9,6.1Hz, 1H),2.16(td,J＝12.9,7.0Hz,1H),2.08–1.99(m,1H),1.85(s,4H)；
HRMS(ESI):m/z calcd for C 19 H 18 NO 4 Cl[M-Cl] + :*,found:*
d2:3.2g, yield 90%; r is R f =0.4 (ethyl acetate/methanol, volume ratio 10:1); 1 H NMR (400MHz,DMSO-d 6 )δ9.33(s,1H),8.57(d,J＝7.7Hz,1H),7.64(t,J＝6.1 Hz,1H),7.26–7.16(m,2H),7.16–7.07(m,3H),6.94(dd,J＝8.2,2.3Hz,1H),6.87–6.80(m,2H),6.76(s,1H),6.54(d,J＝11.4Hz,1H),6.07–5.94(m,1H), 5.94–5.83(m,1H),5.14(dq,J＝17.0,1.5Hz,1H),5.06(ddt,J＝10.0,2.3,1.2Hz,1H),5.04–4.98(m,1H),4.96(ddt,J＝10.0,2.2,1.2Hz,1H),4.37(dt,J＝ 12.0,7.1Hz,1H),3.83(s,3H),3.79(s,3H),3.49(s,3H),3.49–3.43(m,2H),3.40(d,J＝6.9Hz,2H),3.23(d,J＝6.7Hz,2H),2.69(t,J＝7.1Hz,2H), 2.60–2.52(m,1H),2.18(td,J＝12.9,7.1Hz,1H),1.85(s,4H).
d3:3.3g, yield 92%; r is R f =0.4 (ethyl acetate/methanol, volume ratio 10:1); 1 H NMR (400MHz,DMSO-d 6 )δ9.26(s,1H),8.56(d,J＝7.7Hz,1H),7.65(t,J＝6.2 Hz,1H),7.19–7.14(m,2H),7.13–7.07(m,3H),6.90(dd,J＝8.4,2.2Hz,1H),6.81–6.77(m,2H),6.75(s,1H),6.58(d,J＝11.4Hz,1H),5.98(ddt,J＝16.8, 10.0,6.8Hz,1H),5.85(ddt,J＝16.7,10.0,6.7Hz,1H),5.13(dq,J＝17.0,1.5Hz,1H),5.06(ddd,J＝10.0,2.2,1.2Hz,1H),5.01–4.91(m,2H),4.37(dt,J＝ 10.2,7.1Hz,1H),3.82(s,3H),3.78(s,3H),3.47(s,3H),3.39(d,J＝6.8Hz, 2H),3.24–3.16(m,4H),2.56(d,J＝6.3Hz,1H),2.38(t,J＝7.2Hz,2H),1.85(s,4H),1.75(p,J＝7.1Hz,2H).
d4:3.1g, 86% yield; r is R f =0.4 (ethyl acetate/methanol, volume ratio 10:1); 1 H NMR (400MHz,DMSO-d 6 )δ9.24(s,1H),8.56(d,J＝7.7Hz,1H),7.57(t,J＝6.0 Hz,1H),7.19(d,J＝11.1Hz,1H),7.16(dd,J＝8.2,2.2Hz,1H),7.13–7.09(m,2H),7.06(d,J＝8.2Hz,1H),6.93(dd,J＝8.3,2.3Hz,1H),6.85–6.78(m,2H), 6.75(s,1H),6.62(d,J＝11.4Hz,1H),6.07–5.93(m,1H),5.87(ddt,J＝16.8,10.0,6.6Hz,1H),5.13(dq,J＝17.1,1.7Hz,1H),5.06(ddt,J＝10.0,2.3,1.2 Hz,1H),5.03–4.93(m,2H),4.38(dt,J＝12.1,7.1Hz,1H),3.82(s,3H),3.78(s,3H),3.48(s,3H),3.39(d,J＝7.0Hz,2H),3.26(d,J＝6.1Hz,2H),3.21(d,J＝ 6.7Hz,2H),2.61–2.52(m,1H),2.41–2.27(m,2H),2.23–2.12(m,1H), 2.09–1.99(m,1H),1.85(s,4H),1.48(dd,J＝6.7,3.6Hz,4H).
experimental example 1: inhibition of novel coronavirus Main protease (Mpro) by colchicine and magnolol complexes.
The detection is carried out by using a Biyun-Tian kit (P0315M), and a blank group, a control group and a sample group are arranged in a 96-hole blackboard, wherein the detection system is 100 mu L. 96. Mu.L of buffer was added to the blank, 1. Mu.L of the main protease and 95. Mu.L of buffer were added to the control, 1. Mu.L of the main protease, 90. Mu.L of buffer and 5. Mu.LD 1 solution (100. Mu.M), or D2 solution (10. Mu.M, 30. Mu.M, 100. Mu.M), or D3 solution (100. Mu.M), or D4 solution (10. Mu.M, 30. Mu.M, 100. Mu.M) were added to the sample, and incubation was performed at 37℃for 10 minutes. Then, 4. Mu.L of substrate was added to each well, and immediately after mixing, the fluorescence value of each well was measured at Ex/Em=325/393 nm, once every 20 seconds for 30 minutes. The average fluorescence value of each group is calculated and recorded as RFU blank, RFU contrast and RFU sample, and the relative enzyme activity is calculated, wherein the formula is as follows, the enzyme activity= (RFU sample-RFU blank)/(RFU contrast-RFU blank) ×100%
Results: as shown in fig. 1, each of the 4 complexes significantly inhibited the activity of the main protease.
Experimental example 2: inhibition of human neutrophil elastase (neutrophil elastase, NE) by colchicine and magnolol complexes
Abcam kit (ab 118971) is adopted for detection, and a blank group, a control group and a sample group are arranged in a 96-hole blackboard, wherein the detection system is 100 mu L. 75. Mu.L buffer was added to the blank, 50. Mu.L protease and 25. Mu.L buffer were added to the control, 50. Mu.L protease and 25. Mu.L complex solution (10. Mu.M, 30. Mu.M) were added to the sample, and incubation was carried out at 37℃for 5 minutes. Then 25 μl of substrate was added to each well, and immediately after mixing, the fluorescence value of each well was measured at Ex/em=400/505 nm, once every 20 seconds, and the kinetics were observed over 0-40 minutes. In the linear range of the dynamic process, two different time points (T1 and T2) are selected, fluorescence values (RFUT 1 and RFUT 2) corresponding to the time points are obtained, and the slope of the fluorescence values is calculated and can be regarded as the enzyme activity of each hole. Slope= (RFUT 2-RFUT 1)/(T2-T1). Finally, the relative enzyme activity of each group relative to the negative control well was calculated with the enzyme activity of the negative control well being 100%.
Results: as shown in fig. 2, 4 complexes D1, D2, D3, D4 all significantly inhibited neutrophil elastase activity.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (9)
1. A chemical synthesis method of a colchicine and magnolol compound, which is characterized by comprising the following steps:
(1) Mixing colchicine, amino acid, alkali solution and ethanol, and reacting to generate amino acid modified colchicine;
(2) Mixing and reacting amino acid modified colchicine, magnolol, methylene dichloride, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and 4-dimethylaminopyridine to generate colchicine and magnolol compound;
the structural formula of the amino acid is
Wherein n is 2 or 3 or 4;
the chemical structure of the colchicine and magnolol compound is as follows:
wherein n is 2 or 3 or 4.
2. The method according to claim 1, wherein the molar ratio of colchicine to amino acid is 5: (8-12);
the molar volume ratio of colchicine to ethanol is 5mmol: (8-13) mL;
the mass volume ratio of alkali and water in the alkali solution is (0.3-0.6) g:10mL of the alkali solution is sodium hydroxide aqueous solution or potassium hydroxide aqueous solution;
the volume ratio of the alkali solution to the ethanol is 10: (8-13).
3. The process according to claim 2, wherein the reaction in step (1) is carried out at a temperature of 60 to 70 ℃ for a period of 3 to 7 hours.
4. A method according to claim 1 or 3, wherein the molar ratio of colchicine to magnolol is 5 (8-12);
the molar ratio of colchicine to 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride is 5 (8-12);
the molar ratio of colchicine to 4-dimethylaminopyridine is 5: (0.3 to 0.8);
the molar volume ratio of colchicine to dichloromethane is 5mmol: (40-60) mL.
5. The method according to claim 4, wherein the temperature of the reaction in the step (2) is room temperature for 10 to 15 hours.
6. The method according to claim 1, 3 or 5, wherein the product system obtained after the end of the reaction in step (1) is subjected to a purification treatment to obtain pure amino acid modified colchicine for use in step (2);
the purification treatment comprises the following steps:
performing rotary evaporation on the obtained product system to remove ethanol and obtain a solid substance;
washing the obtained solid substance with ethyl acetate, retaining a water phase, adding acid to adjust the pH value to 4-5, and separating out white solid;
washing the obtained white solid with dichloromethane for three times, drying with anhydrous sodium sulfate, and concentrating to obtain pure amino acid modified colchicine;
the volume ratio of the ethyl acetate to the ethanol in the step (1) is 15: (8-13);
the volume ratio of dichloromethane used for each washing and ethanol used in step (1) was independently 30: (8-13).
7. The method of claim 6, wherein the product system is post-treated after the reaction of step (2) to obtain a pure colchicine and magnolol complex;
the post-treatment comprises the following steps:
adding water to quench the reaction, and washing and separating the solution to obtain a water phase;
extracting the obtained water phase with dichloromethane for three times, mixing the organic phases, drying with anhydrous sodium sulfate, concentrating, and subjecting to silica gel column chromatography to obtain pure colchicine and magnolol compound;
the volume ratio of dichloromethane used for each extraction to ethanol used in step (1) was independently 50: (8-13);
the detergent used in the silica gel column chromatography is a mixture of ethyl acetate and methanol, and the volume ratio of the ethyl acetate to the methanol is (8-12): 1.
8. a colchicine and magnolol complex obtained by the method of any one of claims 1 to 7;
the colchicine and magnolol compound has the structure that:
wherein n is 2 or 3 or 4.
9. Use of the colchicine and magnolol complex of claim 8 for preparing an anti-novel coronavirus drug.
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| GB780557A (en) * | 1954-11-03 | 1957-08-07 | Roussel Uclaf | Improvements in or relating to colchicine compounds |
| US5843910A (en) * | 1994-10-05 | 1998-12-01 | Indena S.P.A. | Colchicine derivatives and the therapeutical use thereof |
| WO2012053832A2 (en) * | 2010-10-20 | 2012-04-26 | 주식회사 바이오랜드 | Method for synthesising 4-o-methylhonokiol |
| WO2021052270A1 (en) * | 2019-09-20 | 2021-03-25 | 广东省禾基生物科技有限公司 | Honokiol derivative, preparation method therefor and use thereof |
| CN113181124A (en) * | 2021-05-21 | 2021-07-30 | 劲牌持正堂药业有限公司 | Application of Chinese patent medicine particle composition in preparation of anti-respiratory virus medicine |
| WO2021184128A1 (en) * | 2020-03-20 | 2021-09-23 | Institut De Cardiologie De Montreal | Methods of treating a coronavirus infection using colchicine |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| GB780557A (en) * | 1954-11-03 | 1957-08-07 | Roussel Uclaf | Improvements in or relating to colchicine compounds |
| US5843910A (en) * | 1994-10-05 | 1998-12-01 | Indena S.P.A. | Colchicine derivatives and the therapeutical use thereof |
| WO2012053832A2 (en) * | 2010-10-20 | 2012-04-26 | 주식회사 바이오랜드 | Method for synthesising 4-o-methylhonokiol |
| WO2021052270A1 (en) * | 2019-09-20 | 2021-03-25 | 广东省禾基生物科技有限公司 | Honokiol derivative, preparation method therefor and use thereof |
| WO2021184128A1 (en) * | 2020-03-20 | 2021-09-23 | Institut De Cardiologie De Montreal | Methods of treating a coronavirus infection using colchicine |
| CN113181124A (en) * | 2021-05-21 | 2021-07-30 | 劲牌持正堂药业有限公司 | Application of Chinese patent medicine particle composition in preparation of anti-respiratory virus medicine |
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