CN106361731B - Bibenzyl compound, preparation method thereof and application thereof in preparing antitumor drugs - Google Patents
Bibenzyl compound, preparation method thereof and application thereof in preparing antitumor drugs Download PDFInfo
- Publication number
- CN106361731B CN106361731B CN201610719859.7A CN201610719859A CN106361731B CN 106361731 B CN106361731 B CN 106361731B CN 201610719859 A CN201610719859 A CN 201610719859A CN 106361731 B CN106361731 B CN 106361731B
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- Prior art keywords
- eluent
- ethyl acetate
- ethanol
- water
- compound
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- -1 Bibenzyl compound Chemical class 0.000 title abstract description 14
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- 239000003480 eluent Substances 0.000 claims description 49
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- BXWNKGSJHAJOGX-UHFFFAOYSA-N hexadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCO BXWNKGSJHAJOGX-UHFFFAOYSA-N 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000003906 humectant Substances 0.000 description 1
- 125000001183 hydrocarbyl group Chemical group 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 229920003063 hydroxymethyl cellulose Polymers 0.000 description 1
- 229940031574 hydroxymethyl cellulose Drugs 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 229940049920 malate Drugs 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N malic acid Chemical compound OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- CQDGTJPVBWZJAZ-UHFFFAOYSA-N monoethyl carbonate Chemical compound CCOC(O)=O CQDGTJPVBWZJAZ-UHFFFAOYSA-N 0.000 description 1
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 1
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920005862 polyol Polymers 0.000 description 1
- 150000003077 polyols Chemical class 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 239000003380 propellant Substances 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 150000003856 quaternary ammonium compounds Chemical class 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000008159 sesame oil Substances 0.000 description 1
- 235000011803 sesame oil Nutrition 0.000 description 1
- 150000004760 silicates Chemical class 0.000 description 1
- RMAQACBXLXPBSY-UHFFFAOYSA-N silicic acid Chemical compound O[Si](O)(O)O RMAQACBXLXPBSY-UHFFFAOYSA-N 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 229940095064 tartrate Drugs 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 238000001269 time-of-flight mass spectrometry Methods 0.000 description 1
- UAEJRRZPRZCUBE-UHFFFAOYSA-N trimethoxyalumane Chemical compound [Al+3].[O-]C.[O-]C.[O-]C UAEJRRZPRZCUBE-UHFFFAOYSA-N 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- 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
- A61K31/075—Ethers or acetals
- A61K31/085—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon
-
- 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
- A61K31/075—Ethers or acetals
- A61K31/085—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon
- A61K31/09—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon having two or more such linkages
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C41/00—Preparation of ethers; Preparation of compounds having groups, groups or groups
- C07C41/01—Preparation of ethers
- C07C41/34—Separation; Purification; Stabilisation; Use of additives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C41/00—Preparation of ethers; Preparation of compounds having groups, groups or groups
- C07C41/01—Preparation of ethers
- C07C41/34—Separation; Purification; Stabilisation; Use of additives
- C07C41/36—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C41/00—Preparation of ethers; Preparation of compounds having groups, groups or groups
- C07C41/01—Preparation of ethers
- C07C41/34—Separation; Purification; Stabilisation; Use of additives
- C07C41/38—Separation; Purification; Stabilisation; Use of additives by liquid-liquid treatment
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/898—Orchidaceae (Orchid family)
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses an application of a compound shown as a formula IV or pharmaceutically acceptable salt, crystal form and solvate thereof in preparing a medicament for treating and/or preventing tumors, wherein the compound is as follows: wherein R is1、R4、R6Each independently selected from H or methyl; r2、R3、R5、R7Each independently selected from H or
The invention provides a new application of bibenzyl compounds, in particular to an application in preparing medicaments for treating and/or preventing tumors, which has good inhibition effect on tumor cells, particularly lung cancer cells and IC thereof502-6 mug/mL; meanwhile, the invention also provides a preparation method of the bibenzyl compound.
Description
Technical Field
The invention relates to bibenzyl compounds, a preparation method thereof and application thereof in preparing antitumor drugs.
Background
The prior literature reports that bibenzyl compounds have the drug effect and the application of inhibiting passive skin allergic reaction (passive cutaneous anaphylaxis) and inhibiting β -hexosaminidase released by RBL-2H3 cells (basophilic leukemia cells) (see: Matsuda H et al. antibacterial phenyl anthranes and … Planta Med 2004; 70: 847-:
at present, no research report on the application of the bibenzyl compound in preparing antitumor drugs is found.
Disclosure of Invention
The invention aims to provide a new application of bibenzyl compounds shown as follows, in particular to an application in preparing medicaments for treating and/or preventing tumors.
The invention provides an application of a compound shown in a formula IV or pharmaceutically acceptable salt, crystal form and solvate thereof in preparing a medicament for treating and/or preventing tumors, wherein the compound is as follows:
wherein,
R1、R4、R6each independently selected from H or methyl;
R2、R3、R5、R7each independently selected from H or
The compound shown in the formula IV is:
further, the tumor is cancer; further, the cancer is lung cancer.
The present invention also provides a process for preparing compound F, comprising the steps of:
a. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
b. taking the ethanol extract, and concentrating to obtain a fluid extract;
c. dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, mixing ethyl acetate parts, and recovering solvent to obtain ethyl acetate extract;
d. taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially carrying out gradient elution by using petroleum ether-acetone as an eluent at a ratio of 100:1, at a ratio of 50:1 and at a ratio of 30:1 to obtain petroleum ether: eluent fr.16 when acetone is 30: 1;
e. fr.16 is loaded on a reversed-phase polystyrene resin column, gradient elution is carried out by using ethanol-water of 10:90, 30:70, 50:50, 70:30 and 90:10 as eluent in sequence, and Fr.16-h of the ethanol-water of 90:10 is obtained;
f. fr.16-h is subjected to medium-pressure liquid chromatography, and gradient elution is sequentially carried out by using methanol-water of 20:80, 30:70 and 50:50 as eluent, so as to obtain Fr.16-h-3 when the methanol-water of 50: 50;
g. fr.16-h-3 was chromatographed on silica gel sequentially with dichloromethane-methanol ═ 1: 0. 100, and (2) a step of: 1 as eluent, eluting 4 column volumes respectively, collecting dichloromethane-methanol as 100: fr.16-h-3b at 1;
h. fr.16-h-3 was subjected to gel column chromatography eluting 4 column volumes with methanol-water 70:30 as eluent, followed by preparative thin layer chromatography eluting with dichloromethane-methanol 30:1 as developing agent，RfCompound F was isolated as 0.58.
Further, in the above-mentioned case,
in the step d, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step e, the amount of Fr.16 is 15-20 g, and the gradient elution conditions are as follows:
in the step f, the amount of Fr.16-h is 4-6 g, and the gradient elution conditions are as follows:
the present invention also provides a process for preparing compound G or compound H, comprising the steps of:
i. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
ii. Taking the ethanol extract, and concentrating to obtain a fluid extract;
iii, dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, combining ethyl acetate parts, and recovering the solvent to obtain an ethyl acetate extract;
iv, taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially mixing petroleum ether: gradient elution is carried out by using acetone as an eluent at the ratio of 100:1, 50:1 and 30:1, and petroleum ether is obtained: eluent fr.16 when acetone is 30: 1;
v, fr.16 on a reversed-phase polystyrene type resin column, sequentially mixing ethanol: gradient elution is carried out by using water as eluent, wherein the water is 10:90, 30:70, 50:50, 70:30, 90:10 and 100:0, and ethanol is obtained: fr.16-j when water is 100: 0;
vi, subjecting the Fr.16-j to a sephadex chromatographic column, and sequentially eluting by using dichloromethane-methanol (1: 1) as an eluent, wherein the eluent has the volume of 0.8 time of the column volume and 2.2 times of the column volume to obtain Fr.16-j-1 and Fr.16-j-2 respectively;
vii and fr.16-j-2, silica gel column chromatography, sequentially eluting with dichloromethane-methanol ═ 1: 0. 200: eluting with eluent 1, eluting 3 column volumes respectively, and collecting dichloromethane-methanol (200): fr.16-j-2b at 1;
viii, and Fr.16-j-2b are subjected to sephadex chromatographic column, and petroleum ether-dichloromethane-methanol 2:2:1 is used as eluent, and 1 column volume are sequentially eluted to obtain Fr.16-j-2bA and Fr.16-j-2 bB;
fr.16-j-2bA by preparative thin layer chromatography with dichloromethane-acetone 20:1 as developing solvent, Rf(iv) isolating as 0.76 to give compound G;
fr.16-j-2bB by preparative thin layer chromatography using dichloromethane-acetone 20:1 as developing solvent, Rf0.80, isolated to give compound H.
Further, in the above-mentioned case,
in the step iv, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step v, the amount of Fr.16 is 15-20 g, and gradient elution conditions are as follows:
the present invention also provides a process for preparing compound I, comprising the steps of:
A. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
B. taking the ethanol extract, and concentrating to obtain a fluid extract;
C. dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, mixing ethyl acetate parts, and recovering solvent to obtain ethyl acetate extract;
D. taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially mixing petroleum ether: gradient elution is carried out by using acetone as eluent, wherein the acetone is 100:1, 50:1, 30:1 and 20:1, and petroleum ether is obtained: eluent fr.18 when acetone is 20: 1;
E. fr.18 on a reversed-phase polystyrene type resin column, sequentially eluting with ethanol: gradient elution is carried out by using water as eluent and water as eluent, wherein the water is 40:60, 50:50, 55:45, 60:40, 70:30, 80:20 and 95:5 are carried out to obtain ethanol: fr.18-n at 95: 5;
F. sequentially eluting with dichloromethane-methanol (1: 1) as eluent by Fr.18-n sephadex chromatographic column to obtain Fr.18-n-1, Fr.18-n-2 and Fr.18-n-3 respectively;
fr.18-n-3 was chromatographed on Sephadex column with methanol-water 70:30 as eluent for 3 column volumes, the main compound was chromatographed on preparative thin layer chromatography with dichloromethane-methanol 20:1 as developing solvent, RfThen purified by reverse phase high performance liquid chromatography with methanol-water 65:35 as the mobile phase to give compound I.
Further, in the above-mentioned case,
in the step D, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step E, the amount of Fr.18 is 18-20 g, and the gradient elution conditions are as follows:
the present invention also provides a process for preparing compound J, comprising the steps of:
I. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
II. Taking the ethanol extract, and concentrating to obtain a fluid extract;
III, dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, combining ethyl acetate parts, and recovering the solvent to obtain an ethyl acetate extract;
IV, taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially mixing petroleum ether: gradient elution is carried out by using acetone as eluent, wherein the acetone is 100:1, 50:1, 30:1 and 20:1, and petroleum ether is obtained: eluent fr.18 when acetone is 20: 1;
v, Fr.18 on a reversed-phase polystyrene resin column, sequentially adding ethanol: gradient elution is carried out by using water as eluent and water as eluent, wherein the water is 40:60, 50:50, 55:45, 60:40, 70:30 and 80:20, and ethanol is obtained: fr.18-m at 80: 20;
VI and Fr.18-m are put on a sephadex chromatographic column, and are sequentially eluted by 1 time of column volume and 1 time of column volume by using methanol-water (60: 40) as an eluent to respectively obtain Fr.18-m-1 and Fr.18-m-2;
fr.18-m-2 was chromatographed on Sephadex column with dichloromethane-methanol 1:1 as eluent for 4 column volumes, chromatographed on preparative thin layer chromatography with dichloromethane-methanol 15:1 as developing solvent, RfThen purified by reverse phase high performance liquid chromatography with methanol-water 55:45 as the mobile phase to give compound J.
Further, in the above-mentioned case,
in the step IV, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step V, the amount of Fr.18 is 18-20 g, and the gradient elution conditions are as follows:
the invention provides a new application of bibenzyl compounds, in particular to an application in preparing medicaments for treating and/or preventing tumors, which has good inhibition effect on tumor cells, particularly lung cancer cells and IC thereof502-6 mug/mL; meanwhile, the preparation method of the compound is simple and convenient, mild in reaction conditions, convenient to operate and control, low in energy consumption, high in yield, low in cost and very suitable for industrial production.
The compounds and derivatives provided in the present invention may be named according to the IUPAC (international union of pure and applied chemistry) or CAS (chemical abstracts service, Columbus, OH) naming system.
Definitions of terms used in connection with the present invention: the initial definitions provided herein for a group or term apply to that group or term throughout the specification unless otherwise indicated; for terms not specifically defined herein, the meanings that would be given to them by a person skilled in the art are to be given in light of the disclosure and the context.
"substituted" means that a hydrogen atom in a molecule is replaced by a different atom or molecule.
The minimum and maximum values of the carbon atom content in the hydrocarbon group are indicated by a prefix, e.g. prefix Ca～CbAlkyl means any alkyl group containing "a" to "b" carbon atoms. Thus, for example, C1～C4The alkyl group means an alkyl group containing 1 to 4 carbon atoms, in other words, C1～C4Alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl.
The term "pharmaceutically acceptable" means that the carrier, cargo, diluent, adjuvant, and/or salt formed is generally chemically or physically compatible with the other ingredients comprising a pharmaceutical dosage form and physiologically compatible with the recipient.
The terms "salt" and "pharmaceutically acceptable salt" refer to acid and/or base salts of the above compounds or stereoisomers thereof, with inorganic and/or organic acids and bases, as well as zwitterionic (inner) salts, and also quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly in the final isolation and purification of the compounds. The compound or a stereoisomer thereof may be obtained by appropriately (e.g., equivalently) mixing the above compound or a stereoisomer thereof with a predetermined amount of an acid or a base. These salts may form precipitates in the solution which are collected by filtration, or they may be recovered after evaporation of the solvent, or they may be prepared by reaction in an aqueous medium followed by lyophilization. The salt in the invention can be hydrochloride, sulfate, citrate, benzene sulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compound.
The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.
Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.
Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.
Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like.
In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.
Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.
Dosage forms for topical administration of the compounds of the present invention include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.
The pharmaceutically acceptable auxiliary material of the invention refers to a substance contained in a dosage form except for an active ingredient.
The pharmaceutically acceptable auxiliary components have certain physiological activity, but the addition of the components does not change the dominant position of the pharmaceutical composition in the disease treatment process, but only plays auxiliary effects, and the auxiliary effects are only the utilization of the known activity of the components and are auxiliary treatment modes which are commonly used in the field of medicine. If the auxiliary components are used together with the pharmaceutical composition of the present invention, the protection scope of the present invention still remains.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Detailed Description
The raw materials and equipment used in the embodiment of the present invention are known products and obtained by purchasing commercially available products.
① A Chinese medicinal composition
Bletilla striata medicinal material is purchased from Nenjiang city of Sichuan province at 8 months in 2012, and is identified as a dry tuber of Bletilla striata (Thunb.) Rchb.f. by professor Limin of Chengdu Chinese medicine university.
② reagent and filler
Column chromatography silica gel of 200-300 mesh (reagent grade) purchased from Qingdao ocean silica gel desiccant factory;
thin layer chromatography silica gel G, GF254 and H (chemically pure), purchased from Qingdao ocean silica gel desiccant factory;
MCI gel CHP 20P, 75-150 μm, is a reversed-phase polystyrene-type resin, and is purchased from Mitsubishi chemical corporation of Japan;
sephadex LH-20 sepharose, available from Amersham, Sweden;
a GF254 silica gel preparation thin layer purchased from Yangtze river friend silica gel development Co., Ltd;
chromatographic methanol, 4L/bottle, available from Fisher corporation, USA;
analytically pure reagents such as petroleum ether, ethyl acetate, n-butanol, acetone, methanol and the like are purchased from Chengdu Kelong chemical reagent factories.
③ Experimental instrument
Cometro 6000 high performance liquid chromatography (Cometro, usa);
waters Synapt G2HDMS high resolution time-of-flight mass spectrometry (Waters, usa);
nuclear magnetic resonance apparatus: bruker-600 NMR; measuring temperature: 297K; taking a solvent peak signal as a reference;
vector 22FT-IR infrared spectrometer (Bruker, Switzerland);
shimadzu UV-260 UV-visible spectrophotometer (Shimadzu, japan);
BP211D one tenth ten million electronic balance (Sartorius, switzerland);
r-210 rotary evaporator (BUCHI, Switzerland);
model DZG-6050 vacuum drying oven (shanghai semen).
Example 1 extraction and Structure identification of Compound F
(1) Extracting medicinal materials:
extracting dried rhizoma Bletillae coarse powder (3kg) with 95% ethanol under reflux for 3 times (30L × 3), each time for 2 hr;
(2) separation and purification of components:
①, concentrating the ethanol extract (90L) under reduced pressure, and drying to obtain semisolid fluid extract 510 g;
② dispersing the semisolid fluid extract (500g) with water (5L), sequentially extracting with ethyl acetate and n-butanol (20L), mixing ethyl acetate parts, and recovering solvent under reduced pressure to obtain 160g of ethyl acetate extract;
③ separating ethyl acetate extract (160g) by silica gel column chromatography, gradient eluting with petroleum ether and acetone, and detecting the eluate by thin layer chromatography to obtain eluate Fr.16 when the ratio of petroleum ether and acetone is 30: 1;
the conditions for gradient elution were as follows:
④, subjecting Fr.16 fraction (18g) to MCI treatment, gradient eluting with 10% -100% v/v ethanol water, and detecting by thin layer chromatography to obtain 10 fractions Fr.16-a-Fr.16-j;
the conditions for gradient elution were as follows:
⑤, subjecting Fr.16-h component (5.0g) to medium pressure liquid chromatography (B-chi Gradient Former B-687 medium pressure liquid chromatograph) with Gradient elution of 20%, 30%, 50%, 70%, 80%, 95%, 100% methanol water, eluting 2, 3, 4, 5 column volumes respectively to obtain seven components Fr.16-h-1-Fr.16-h-7;
⑥, subjecting Fr.16-h-3 component (800mg) to silica gel column chromatography, eluting with dichloromethane-methanol 1: 0, 100:1, 50:1 at each ratio for 4 column volumes, and collecting the fraction Fr.16-h-3b eluted at 100: 1;
⑦ and Fr.16-h-3b (140mg) are subjected to gel column chromatography, 70% methanol water is used as an eluent to elute 4 column volumes, and then compound F is separated by preparative thin layer chromatography (a developing solvent is dichloromethane-methanol-30: 1, Rf is 0.58);
when the bibenzyl compounds in the rhizoma bletillae are developed by thin-layer chromatography, 10% sulfuric acid ethanol test solution is sprayed, and the rhizoma bletillae is developed for 3min at 105 ℃ to show more yellow, orange to reddish spots, so that the rhizoma bletillae can be used for tracking detection of bibenzyl;
mass spectral data for compound F: ESI-MS M/z 349[ M-H]-；
Hydrogen spectrum data:1H NMR(CD3COCD3,600MHz)δ:7.06(1H,t,J＝7.8Hz,H-5′),6.93(2H,d,J＝8.4Hz,H-2″,6″),6.68(2H,d,J＝8.4Hz,H-3″,5″),6.66(1H,t,J＝1.8Hz,H-2′),6.64(1H,dd,J＝7.8,1.8Hz,H-4′),6.62(1H,dd,J＝7.8,1.8Hz,H-2′),6.40(1H,d,J＝2.4Hz,H-4),6.38(1H,d,J＝2.4Hz,H-6),3.89(2H,s,CH2-γ),3.75(3H,s,OCH3-3),2.59(2H,m,CH2-α),2.74(2H,m,CH2-β)；
carbon spectrum data:13C NMR(CD3COCD3,150MHz)δ:160.2(C-3),158.7(C-3′),158.0(C-5),156.5(C-4″),145.0(C-1′),143.7(C-1),134.0(C-1″),130.6(C-5′),130.3(C-2″,6″),120.8(C-6′),119.8(C-2),116.6(C-4′),116.2(C-3″,5″),114.1(C-2′),109.5(C-6),98.3(C-4),56.3(C-OMe-3),38.6(C-β),36.5(C-α),30.7(C-γ)；
from the above data, compound F was identified as 2-p-hydroxybenzyl-3', 5-dihydroxy-3-methoxybenzyl, and its structure was as follows:
example 2 extraction and Structure identification of Compound G
(1) Extracting medicinal materials:
extracting dried rhizoma Bletillae coarse powder (3kg) with 95% ethanol under reflux for 3 times (30L × 3), each time for 2 hr;
(2) separation and purification of components:
①, concentrating the ethanol extract (90L) under reduced pressure, and drying to obtain semisolid fluid extract 510 g;
② dispersing the semisolid fluid extract (500g) with water (5L), sequentially extracting with ethyl acetate and n-butanol (20L), mixing ethyl acetate parts, and recovering solvent under reduced pressure to obtain 160g of ethyl acetate extract;
③ separating ethyl acetate extract (160g) by silica gel column chromatography, gradient eluting with petroleum ether and acetone, and detecting the eluate by thin layer chromatography to obtain eluate Fr.16 when the ratio of petroleum ether and acetone is 30: 1;
the conditions for gradient elution were as follows:
④, subjecting Fr.16 fraction (18g) to MCI treatment, gradient eluting with 10% -100% v/v ethanol water, and detecting by thin layer chromatography to obtain 10 fractions Fr.16-a-Fr.16-j;
the conditions for gradient elution were as follows:
⑤, passing Fr.16-j (3.0g) through gel LH-20 chromatographic column, eluting with dichloromethane-methanol (1: 1) as eluent, wherein Fr.16-j-1, Fr.16-j-2 and Fr.16-j-3 are respectively collected when eluting 0.8 times of column volume, 2.2 times of column volume and 3 times of column volume, and Fr.16-j-2 is the concentrated part of bibenzyl compound;
⑥ and Fr.16-j-2 (1.8g) are respectively eluted by dichloromethane-methanol 1: 0, 200:1, 100:1 and 50:1 through silica gel column chromatography, 3 column volumes of eluent in each proportion are eluted, the solvent is recovered under reduced pressure, and an elution part Fr.16-j-2b of 200:1 is collected;
⑦ and Fr.16-j-2b fraction (0.8g) passing through gel LH-20 chromatographic column, eluting with petroleum ether-dichloromethane-methanol 2:2:1 as eluent, sequentially eluting for 1 and 1 column volumes to obtain Fr.16-j-2bA and Fr.16-j-2bB fractions, and subjecting Fr.16-j-2bA to thin layer chromatography (developing solvent dichloromethane-acetone 20:1, R is used as developing solvent)f0.76) to yield compound G;
when the bibenzyl compounds in the rhizoma bletillae are developed by thin-layer chromatography, 10% sulfuric acid ethanol test solution is sprayed, and the rhizoma bletillae is developed for more than yellow, orange to reddish spots at 105 ℃ for 3min, so that the rhizoma bletillae can be used for tracking detection of bibenzyl;
mass spectral data for compound G: ESI-MS M/z 363[ M-H ]]-；
Hydrogen spectrum data:1H NMR(CD3COCD3,600MHz)δ:7.15(1H,t,J＝7.8Hz,H-5′),7.01(2H,d,J＝8.4Hz,H-2″,6″),6.74(1H,brd,J＝7.8Hz,H-4′),6.72(1H,brd,J＝7.8Hz,H-6′)6.71(2H,d,J＝8.4Hz,H-3″,5″),6.70(1H,brs,H-2′),6.41(1H,d,J＝2.4Hz,H-4),6.38(1H,d,J＝2.4Hz,H-6),3.95(2H,s,CH2-γ),3.75(3H,s,OCH3-3′),3.71(3H,s,OCH3-5),2.79(2H,m,CH2-α),2.65(2H,m,CH2-β)；
from the above data, compound G was identified as 2-p-hydroxybenzyl-3-hydroxy-3', 5-dimethoxybibenzyl, having the following structure:
example 3 extraction and Structure identification of Compound H
(1) Extracting medicinal materials:
extracting dried rhizoma Bletillae coarse powder (3kg) with 95% ethanol under reflux for 3 times (30L × 3), each time for 2 hr;
(2) separation and purification of components:
①, concentrating the ethanol extract (90L) under reduced pressure, and drying to obtain semisolid fluid extract 510 g;
② dispersing the semisolid fluid extract (500g) with water (5L), sequentially extracting with ethyl acetate and n-butanol (20L), mixing ethyl acetate parts, and recovering solvent under reduced pressure to obtain 160g of ethyl acetate extract;
③ separating ethyl acetate extract (160g) by silica gel column chromatography, gradient eluting with petroleum ether and acetone, and detecting the eluate by thin layer chromatography to obtain eluate Fr.16 when the ratio of petroleum ether and acetone is 30: 1;
the conditions for gradient elution were as follows:
④, subjecting Fr.16 fraction (18g) to MCI treatment, gradient eluting with 10% -100% v/v ethanol water, and detecting by thin layer chromatography to obtain 10 fractions Fr.16-a-Fr.16-j;
the conditions for gradient elution were as follows:
⑤, passing Fr.16-j (3.0g) through gel LH-20 chromatographic column, eluting with dichloromethane-methanol (1: 1) as eluent, wherein Fr.16-j-1, Fr.16-j-2 and Fr.16-j-3 are respectively collected when eluting 0.8 times of column volume, 2.2 times of column volume and 3 times of column volume, and Fr.16-j-2 is the concentrated part of bibenzyl compound;
⑥ and Fr.16-j-2 (1.8g) are respectively eluted by dichloromethane-methanol 1: 0, 200:1, 100:1 and 50:1 through silica gel column chromatography, 3 column volumes of eluent in each proportion are eluted, the solvent is recovered under reduced pressure, and an elution part Fr.16-j-2b of 200:1 is collected;
⑦ and Fr.16-j-2b fraction (0.8g) passing through gel LH-20 chromatographic column, eluting with petroleum ether-dichloromethane-methanol 2:2:1 as eluent, sequentially eluting for 1 and 1 column volumes to obtain Fr.16-j-2bA and Fr.16-j-2bB fractions, and subjecting Fr.16-j-2bB fraction to thin layer chromatography (developing solvent dichloromethane-acetone 20:1, R)f0.80) to yield compound H;
when the bibenzyl compounds in the rhizoma bletillae are developed by thin-layer chromatography, 10% sulfuric acid ethanol test solution is sprayed, and the rhizoma bletillae is developed for more than yellow, orange to reddish spots at 105 ℃ for 3min, so that the rhizoma bletillae can be used for tracking detection of bibenzyl;
mass spectral data for compound H: ESI-MS M/z 363[ M-H ]]-；
Hydrogen spectrum data:1H NMR(CD3COCD3,600MHz)δ:7.17(1H,t,J＝7.8Hz,H-5′),7.10(2H,d,J＝8.4Hz,H-2″,6″),6.79(1H,d,J＝7.8Hz,H-6′),6.76(1H,brs,H-2′),6.73(1H,dd,J＝7.8,1.8Hz,H-4′),6.66(2H,d,J＝8.4Hz,H-3″,5″),6.41(1H,brs,H-6),6.38(1H,brs,H-2),3.84(2H,s,CH2-γ),3.75(3H,s,OCH3-3′),3.72(3H,s,OCH3-3),2.86(2H,m,CH2-α),2.79(2H,m,CH2-β)；
carbon spectrum data:13C NMR(CD3COCD3,150MHz)δ:160.6(C-3′),159.3(C-3),156.3(C-5),155.9(C-4″),144.4(C-1′),141.8(C-1),133.7(C-1″),130.3(C-2″,6″),130.0(C-5′),121.5(C-6′),115.4(C-3″,5″),114.9(C-2′),114.9(C-4),112.1(C-4′),109.2(C-6),103.6(C-2),55.8(OCH3-3′),55.3(OCH3-3),38.6(CH2-α),38.4(CH2-β),28.3(CH2-γ)；
from the above data, compound H was determined to be 4-p-hydroxybenzyl-5-hydroxy-3, 3' -dimethoxybibenzyl, having the following structure:
example 4 extraction and Structure identification of Compound I
(1) Extracting medicinal materials:
extracting dried rhizoma Bletillae coarse powder (3kg) with 95% ethanol under reflux for 3 times (30L × 3), each time for 2 hr;
(2) separation and purification of components:
①, concentrating the ethanol extract (90L) under reduced pressure, and drying to obtain semisolid fluid extract 510 g;
② dispersing the semisolid fluid extract (500g) with water (5L), sequentially extracting with ethyl acetate and n-butanol (20L), mixing ethyl acetate parts, and recovering solvent under reduced pressure to obtain 160g of ethyl acetate extract;
③ separating ethyl acetate extract (160g) by silica gel column chromatography, gradient eluting with petroleum ether and acetone, detecting the eluate by thin layer chromatography, and mixing similar components to obtain eluate Fr.18 at a ratio of 20: 1;
the conditions for gradient elution were as follows:
④ Fr.18 fraction (19g) was MCI treated, eluted with 40% -100% ethanol water, collected according to thin layer tracing
Recovering solvent from the eluate containing the target compound to obtain 15 fractions Fr.18-a to Fr.18-o;
⑤, passing Fr.18-n fraction (95% ethanol elution part, 0.7g) through gel LH-20, eluting with dichloromethane-methanol 1:1 as eluent, sequentially collecting fractions for Fr.18-n-1, Fr.18-n-2, and Fr.18-n-3 eluting 1.5 times, 3 times, and 5 times of column volume respectively;
subjecting Fr.18-n-3 (300mg) to gel LH-20 column chromatography with 70% methanol as eluent, eluting for 3 column volumes, subjecting the main compound to preparative thin layer chromatography (dichloromethane-methanol 20:1 as developing solvent, Rf0.78) and reversed-phase high performance liquid chromatography (65% methanol water is used as a mobile phase) to prepare and purify the compound I;
when the bibenzyl compounds in the rhizoma bletillae are developed by thin-layer chromatography, 10% sulfuric acid ethanol test solution is sprayed, and the rhizoma bletillae is developed for more than yellow, orange to reddish spots at 105 ℃ for 3min, so that the rhizoma bletillae can be used for tracking detection of bibenzyl;
mass spectral data for compound I: ESI-MS M/z 469[ M-H]-；
Hydrogen spectrum data:1H NMR(CD3COCD3,600MHz)δ:7.14(1H,t,J＝7.8Hz,H-5′),7.01(2H,d,J＝8.4Hz,H-H-2″,6″),6.94(2H,d,J＝8.4Hz,H-H-2′,6′),6.67-6.71(6H,m,H-4′,6′,3′,5′,3″,5″),6.63(1H,brs,H-2′),6.61(1H,s,H-4),4.03(2H,s,CH2-γ1),3.98(2H,s,CH2-γ2),3.76(3H,s,OCH3-3′),3.75(3H,s,OCH3-3),2.76(2H,m,CH2-α),2.39(2H,m,-CH2-β)；
from the above data, compound I was identified as 2, 6-bis (p-hydroxybenzyl) -5-hydroxy-3, 3' -dimethoxybibenzyl, having the following structure:
example 5 extraction and Structure identification of Compound J
(1) Extracting medicinal materials:
extracting dried rhizoma Bletillae coarse powder (3kg) with 95% ethanol under reflux for 3 times (30L × 3), each time for 2 hr;
(2) separation and purification of components:
①, concentrating the ethanol extract (90L) under reduced pressure, and drying to obtain semisolid fluid extract 510 g;
② dispersing the semisolid fluid extract (500g) with water (5L), sequentially extracting with ethyl acetate and n-butanol (20L), mixing ethyl acetate parts, and recovering solvent under reduced pressure to obtain 160g of ethyl acetate extract;
③ separating ethyl acetate extract (160g) by silica gel column chromatography, gradient eluting with petroleum ether and acetone, detecting the eluate by thin layer chromatography, and mixing similar components to obtain eluate Fr.18 at a ratio of 20: 1;
the conditions for gradient elution were as follows:
④, Fr.18 fraction (19g) is processed by MCI, eluted with 40% -100% ethanol water, and the eluate containing the target compound is collected according to thin layer tracing, and the solvent is recovered to obtain 15 components Fr.18-a-Fr.18-o;
⑤ and Fr.18-m components (80% ethanol elution part, 0.9g) are separated by a gel LH-20 chromatographic column, 60% methanol water is used as an eluent to elute 1, 1 and 1 column volumes in sequence, and six components Fr.18-m-1 to Fr.18-m-6 are respectively obtained in sequence;
subjecting Fr.18-m-2 fraction to gel LH-20 column chromatography (eluent dichloromethane-methanol 1:1), eluting for 4 column volumes, and subjecting to preparative thin layer chromatography (developing solvent dichloromethane-methanol 15:1, R)f0.68) and reverse phase high performance liquid chromatography with 55% methanol water to obtain compound J;
when the bibenzyl compounds in the rhizoma bletillae are developed by thin-layer chromatography, 10% sulfuric acid ethanol test solution is sprayed, and the rhizoma bletillae is developed for 3min at 105 ℃, so that yellow, orange-red to reddish spots are developed, and the rhizoma bletillae can be used for tracking detection of bibenzyl;
mass spectral data for compound J: ESI-MS M/z 561[ M-H ]]-；
Hydrogen spectrum data1H NMR(CD3COCD3,600MHz)δ:7.04(2H,d,J＝8.4Hz,H-2″″,6″″),6.99(2H,d,J＝8.4Hz,H-2″,6″),6.92(2H,d,J＝8.4Hz,H-2″′,6″′),6.88(1H,d,J＝9.0Hz,H-5′),6.71(2H,d,J＝8.4Hz,H-3″″,5″″),6.69(1H,brs,H-2′),6.67(2H,d,J＝8.4Hz,H-3″,5″),6.66(2H,d,J＝8.4Hz,H-3″′,5″′),6.58(1H,s,H-4),6.52(1H,dd,J＝9.0,1.8Hz,H-6′),3.99(2H,s,CH2-7″),3.94(2H,s,CH2-7″′),3.80(2H,s,CH2-7″″),3.75(3H,s,OCH3),2.73(2H,m,CH2-α),2.33(2H,m,CH2-β)；
Carbon spectrum data:13C NMR(CD3COCD3,150MHz)δ:157.8(C-3),156.2(C-4″″),156.0(C-4″),155.9(C-4″′),155.5(C-3′),155.4(C-5),142.3(C-1),142.3(C-1′),133.6(C-1″′),133.6(C-1″),133.1(C-1″″),131.1(C-5′),130.6(C-2″″,6″″),129.9(C-2″,6″),129.7(C-2″′),126.7(C-4′),120.2(C-6′),119.9(C-2),119.0(C-6),115.7(C-3″,5″；C-3″′,5″′；C-3″″,5″″),115.5(C-2′),97.8(C-4),55.7(C-OCH3),37.0(C-β),35.1(C-7″″),33.3(C-α),30.9(C-7″),30.8(C-7″′)；
from the above data, compound J is 2,4 ', 6-tris (p-hydroxybenzyl) -3', 5-dihydroxy-3-methoxybibenzyl, having the following structure:
the advantageous effects of the present invention will be specifically described below by way of test examples.
Example 6 use of Compounds F to J according to the invention for the treatment and/or prophylaxis of tumors
(1) Experimental materials:
① cells
Lung cancer cell a549 was derived from ATCC company, usa.
② Experimental instrument
HWCL-1 constant temperature magnetic stirring bath (Zhengzhou great wall science and trade Co., Ltd.);
BP211D one tenth ten million electronic balance (Sartorius, switzerland);
model DZG-6050 vacuum drying oven (shanghai semen);
microplate reader (Thermo 3001, Thermo Fisher Scientific);
an electronic balance (model ESJ120-4, shenyang dragon electronic weighing instrument ltd);
ultra clean bench (MCV-B161F (T), SANYO, Japan);
microscope (Primo Vert, AxioCam ERc 5s, ZEISS);
CO2incubator (HH.CP-T, Shanghai Zixin scientific instruments Co., Ltd.).
(2) The experimental method comprises the following steps:
① inoculation of cells
Cells in the logarithmic growth phase were digested with 0.25% pancreatin. The single cell suspension is prepared by culturing the cells in a cell culture medium containing 10% FBS. Counting by using a cell counting plate, inoculating A549 tumor cells in a good state to a 96-well plate to ensure that the cell density is 4 multiplied by 103Each well was filled with 100. mu.L of cell suspension, incubated at 37 ℃ with 5% CO2Culturing in an incubator for 24 h.
② drug treatment
Starting from 20. mu.g/mL of each sample, the samples were diluted with a gradient of medium, 2-fold diluted, 5 drug concentrations were set, and duplicate wells were tested for each concentration. The drug is added into each hole with the concentration of 20, 10, 5, 2.5 and 1.25 mug/mL for 100 mug L, each concentration is provided with 3 multiple holes, and the steps are repeated for 3 times. The negative control group is a culture medium solution containing a proper amount of DMSO, and the blank control group is a culture medium and a solvent which do not contain cells. The 96-well plate was returned to the incubator and exposed for 72h at 37 ℃.
③ color development and IC50Is calculated by
After 72h, 20. mu.L of MTT solution (5g/L) was added to each well in the dark, incubation was continued for 4h, the supernatant was discarded, 150. mu.L of DMSO was added to each well, the mixture was placed on a shaker and shaken at a slow speed for 10min to dissolve formazan sufficiently, and then OD at a wavelength of 570nm was measured using a microplate reader.
IC by using improved Kouya method50＝log-1[Xm-i(∑P-0.5)]A calculation was performed wherein Xm: a designed logarithm of maximum concentration; i: log of each concentration multiple concentration; e, sigma P: sum of growth inhibition rates of each group; 0.5: an empirical constant. By repeating the experiment three times, 3 ICs were calculated50Values (two decimal places are reserved), and the average value is calculated; the test results are shown in Table 1.
TABLE 1 cytotoxic Activity IC of bibenzyl Compounds F-J of the present invention on A549 tumor cells50
| NO. | IC50(μg/mL) |
| Compound F | 5.13 |
| Compound G | 5.88 |
| Compound H | 4.83 |
| Compound I | 3.80 |
| Compound J | 2.01 |
The results show that the bibenzyl compound has good inhibition effect on tumor cells and can be used for preparing antitumor drugs.
In conclusion, the invention provides a new application of bibenzyl compounds, in particular to an application in preparing medicaments for treating and/or preventing tumors, which has good inhibition effect on tumor cells, particularly on lung cancer cells and IC thereof502-6 mug/mL; meanwhile, the preparation method of the compound is simple and convenient, mild in reaction conditions, convenient to operate and control, low in energy consumption, high in yield, low in cost and very suitable for industrial production.
Claims (5)
1. The application of the following compounds or pharmaceutically acceptable salts thereof in preparing medicines for treating and/or preventing lung cancer:
2. use according to claim 1, characterized in that: the preparation method of the compound G or the compound H comprises the following steps:
i. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
ii. Taking the ethanol extract, and concentrating to obtain a fluid extract;
iii, dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, combining ethyl acetate parts, and recovering the solvent to obtain an ethyl acetate extract;
iv, taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially mixing petroleum ether: gradient elution is carried out by using acetone as an eluent at the ratio of 100:1, 50:1 and 30:1, and petroleum ether is obtained: eluent fr.16 when acetone is 30: 1;
v, fr.16 on a reversed-phase polystyrene type resin column, sequentially mixing ethanol: gradient elution is carried out by using water as eluent, wherein the water is 10:90, 30:70, 50:50, 70:30, 90:10 and 100:0, and ethanol is obtained: fr.16-j when water is 100: 0;
vi, subjecting the Fr.16-j to a sephadex chromatographic column, and sequentially eluting by using dichloromethane-methanol (1: 1) as an eluent, wherein the eluent has the volume of 0.8 time of the column volume and 2.2 times of the column volume to obtain Fr.16-j-1 and Fr.16-j-2 respectively;
vii and fr.16-j-2, silica gel column chromatography, sequentially eluting with dichloromethane-methanol ═ 1: 0. 200: eluting with eluent 1, eluting 3 column volumes respectively, and collecting dichloromethane-methanol (200): fr.16-j-2b at 1;
viii, and Fr.16-j-2b are subjected to sephadex chromatographic column, and petroleum ether-dichloromethane-methanol 2:2:1 is used as eluent, and 1 column volume are sequentially eluted to obtain Fr.16-j-2bA and Fr.16-j-2 bB;
fr.16-j-2bA by preparative thin layer chromatography with dichloromethane-acetone 20:1 as developing solvent, Rf(iv) isolating as 0.76 to give compound G;
fr.16-j-2bB by preparative thin layer chromatography using dichloromethane-acetone 20:1 as developing solvent, Rf0.80, isolated to give compound H.
3. Use according to claim 2, characterized in that:
in the step iv, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step v, the amount of Fr.16 is 15-20 g, and gradient elution conditions are as follows:
4. use according to claim 1, characterized in that: the preparation method of the compound I comprises the following steps:
A. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
B. taking the ethanol extract, and concentrating to obtain a fluid extract;
C. dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, mixing ethyl acetate parts, and recovering solvent to obtain ethyl acetate extract;
D. taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially mixing petroleum ether: gradient elution is carried out by using acetone as eluent, wherein the acetone is 100:1, 50:1, 30:1 and 20:1, and petroleum ether is obtained: eluent fr.18 when acetone is 20: 1;
E. fr.18 on a reversed-phase polystyrene type resin column, sequentially eluting with ethanol: gradient elution is carried out by using water as eluent and water as eluent, wherein the water is 40:60, 50:50, 55:45, 60:40, 70:30, 80:20 and 95:5 are carried out to obtain ethanol: fr.18-n at 95: 5;
F. sequentially eluting with dichloromethane-methanol (1: 1) as eluent by Fr.18-n sephadex chromatographic column to obtain Fr.18-n-1, Fr.18-n-2 and Fr.18-n-3 respectively;
fr.18-n-3 was chromatographed on Sephadex column with methanol-water 70:30 as eluent for 3 column volumes, the main compound was chromatographed on preparative thin layer chromatography with dichloromethane-methanol 20:1 as developing solvent, RfThen purified by reverse phase high performance liquid chromatography with methanol-water 65:35 as the mobile phase to give compound I.
5. Use according to claim 4, characterized in that:
in the step D, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step E, the amount of Fr.18 is 18-20 g, and the gradient elution conditions are as follows:
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| CN201610719859.7A Expired - Fee Related CN106361731B (en) | 2016-08-24 | 2016-08-24 | Bibenzyl compound, preparation method thereof and application thereof in preparing antitumor drugs |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103933350A (en) * | 2012-12-31 | 2014-07-23 | 成都中医药大学 | Use of rhizoma bletillae ethyl acetate extract |
| CN105001064A (en) * | 2015-07-10 | 2015-10-28 | 浙江中医药大学 | Bletilla striata fibrous root blestriarene A and preparation method and application thereof |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103933350A (en) * | 2012-12-31 | 2014-07-23 | 成都中医药大学 | Use of rhizoma bletillae ethyl acetate extract |
| CN105001064A (en) * | 2015-07-10 | 2015-10-28 | 浙江中医药大学 | Bletilla striata fibrous root blestriarene A and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| "Novel Bibenzyl Derivatives from the Tubers of Bletilla striata";Jia-Quan Feng, 等;《Helvetica Chimica Acta》;20081231;第91卷(第3期);520-525 * |
| "石斛中联苄类化合物抗血管新生活性研究";沈凯凯，等;《时珍国医国药》;20111231;第22卷(第4期);919-921页 * |
| Antimitotic activity and reversal of breast cancer resistance protein-mediated drug resistance by stilbenoids from Bletilla striata;Hiroshi Morita 等;《Bioorganic & Medicinal Chemistry Letters》;20041229;1051–1054页 * |
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