CN116041307A - Preparation method of 4, 9-dihydroxyl-alpha-lapachone from catalpa bungei - Google Patents
Preparation method of 4, 9-dihydroxyl-alpha-lapachone from catalpa bungei Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/78—Ring systems having three or more relevant rings
- C07D311/92—Naphthopyrans; Hydrogenated naphthopyrans
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/07—Optical isomers
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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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Abstract
The invention relates to a method for preparing 4, 9-dihydroxyl-containing material from catalpa bungeiαThe preparation method of the lapachone comprises the steps of placing the golden silk tree into dichloromethane for cold leaching, filtering to obtain an extracting solution, and carrying out reduced pressure distillation to obtain dark brown extract; loading the obtained extract onto silica gel column, eluting with mixed solution of petroleum ether and ethyl acetate to obtain red crystal, and detecting to obtain 4, 9-dihydroxyl-containing productαLapachone with a purity of more than 99%. The technology provided by the invention has the advantages of high yield, simple steps and easy industrialized production; 4, 9-dihydroxyl-containing compositions prepared by the present inventionαLapachone can be further prepared as an intermediate by chiral column separation (4S) 4, 9-dihydroxyl-α-lapachone and (4)R) 4, 9-dihydroxyl-αLapaquinone can also be used as medicine source medicine for preparing antineoplastic medicine.
Description
Technical Field
The invention relates to the fields of chemical engineering and natural medicines, in particular to a method for preparing 4, 9-dihydroxyl-containing material in a catalpa bungeiα-a process for the preparation of lapachone.
Background
Sorbus pohuashanensis (Thunb.) MakinoCatalpa bungei) Is of Bignoniaceae familyBignoniaceae) Catalpa alta (L.) SprengCatalpa) The plant is the important country in ChinaThe tree species is a special precious material and ornamental tree species, and the cultivation history is long, and the tree species has the reputation of 'wood king'. In China, the east is the east coast, the west is the Gansu orchid state, the Tianshui and the Sichuan han source, the north is the Shandong customs, the south is the cloud to the Lincang and the Guangzhou in Guangdong are distributed by catalpa bungei, and the land spans 3 vegetation areas of the grassland area, the warm-area deciduous broadleaf forest area and the subtropical evergreen broadleaf forest area.
The application of the catalpa bungei is wide, besides being used for landscaping and plate manufacturing, the medical application of the catalpa bungei is introduced in Chinese traditional medicine resource will: its root bark and bark (bark of catalpa bungei), bitter and cool. Clearing away heat and toxic materials, dispelling blood stasis and relieving swelling. Can be used for treating traumatic injury, fracture, carbuncle, sore, toxic swelling, hemorrhoid, fistula, vomiting, and cough. Its leaf (catalpa bungei leaf), bitter and cool. Detumescence, pus discharge and granulation promotion. Can be used for treating tumor and scrofula. It is bitter and cool. Detoxify, relieve pain and promote tissue regeneration. Its fruits and seeds are bitter and cool. Clear heat and induce diuresis. Can be used for treating stranguria, stranguria with urinary calculus, toxic heat, and scabies.
In recent 10 years, chinese scholars have considerable literature reports on a plurality of fields such as catalpa bungei breeding technology, planting resource research, growth and development research, cultivation and forestation technology, plant diseases and insect pests research, genetic breeding, variety breeding and application and the like, and have fresh reports on chemical components and biological activities. The research on catalpa bungei is also focused on flowers, leaves, fruits, seeds and the like, and the related research on heartwood is almost not available, and particularly, the research on chemical components, medicinal activity and related mechanisms of the heartwood is rarely reported. Kalopanax pictus nakaiCatalpa bungei"Jisi") is an important member of the catalpa tree group, although the presence of the antitumor active substance naphthoquinone compounds in plants of this genus has been reported in the literature (Fisterapia, 2022, 160, 105196), wherein 4, 9-dihydroxyl-αLapachone and chiral components thereof have obvious effect of inhibiting proliferation activity (IC) of human breast cancer cell strain MCF-7 50 =2.19 μm), which is equivalent to the clinical drug β -lapachone activity, but the extraction and preparation methods are cumbersome, require multi-stage column chromatography and semi-preparative liquid phase purification, and have low overall yield (0.24%), which is not conducive to large-scale preparation.
The antitumor activity of naphthoquinone compounds has long been concerned by scholars at home and abroadSuch as violaxoids, juglone, salvainβSome of them have been subjected to preclinical studies. The mechanism of the anti-tumor effect can interfere with a plurality of enzyme systems which are necessary for the growth of tumor besides the alkylating effect on tumor cells reported earlier, such as topoisomerase, telomerase, caspase, protein-tyrosine phosphorylase and the like, induce apoptosis and show different selectivities for different enzyme systems.
Therefore, the invention takes the catalpa bungei as the material and provides the chemical component 4, 9-dihydroxyl-derived from the heartwoodαThe preparation method of the lapachone has high yield and simple steps, and is easy for industrial production. The method provides a new idea for developing, protecting and breeding the catalpa bungei and other catalpa bungei members in China, and expands the research path of the natural product source antitumor lead medicine.
Disclosure of Invention
The invention aims at solving the problems that no clear method is available in the prior art for extracting and preparing the 4, 9-dihydroxyl-containing material in the golden threadαLapatiquinone, provides a 4, 9-dihydroxy-in Sorbus bungeanaα-a process for the preparation of lapachone.
The invention provides a catalpa bungei 4, 9-dihydroxyl-α-a process for the preparation of lapachone, comprising the steps of:
(1) Pulverizing Aronia melanocarpa, cold soaking in dichloromethane solvent with several times of the powder, filtering to obtain extractive solution, and vacuum distilling to obtain extract.
(2) Mixing 200-300 mesh silica gel with petroleum ether, and loading into chromatographic column.
(3) Dissolving the extract in the step (1) by using dichloromethane, mixing the extract with silica gel with the mass of several times of the extract, loading the sample by a dry method (or mixing the extract with petroleum ether with the same volume, loading the sample by a wet method), and using petroleum ether after loading: eluting with ethyl acetate mixed solvent, detecting by TLC thin layer chromatography, and detecting according to R f Segmenting the value and color, combining R f Concentrating under reduced pressure, drying to obtain red crystal, and detecting to obtain 4, 9-dihydroxyl-α-LapaQuinone.
The soaking temperature in the step (1) is normal temperature, and the soaking time is 70-74 h.
The volume ratio of petroleum ether to ethyl acetate in the step (3) is 10-15:1.
said R is f The value is 0.16, and the developing agent is petroleum ether: ethyl acetate mixed solventv/v, 10:1）。
Prepared 4, 9-dihydroxyl-αThe purity of the lapachone detected by the AQ-C18 high performance liquid chromatography is more than 99 percent.
AQ-C18 high performance liquid chromatography conditions: the chromatographic column is AQ-C18 column 4.6X1250 mm,5 μm; the mobile phase is methanol (A phase) -water (B phase) (gradient: 0-20 min, 10-100% methanol, 20-30 min,100% methanol, 30-35 min, 100-10% methanol); the flow rate is 1.0 mL/min; the sample injection amount is 10 mu L; the detection wavelength is 254 nm; the column temperature was 30 ℃.
The 4, 9-dihydroxyl-containing material prepared by the methodαLapachone, 4, 9-dihydroxyl-αThe lapachone comprises (4)R) 4, 9-dihydroxyl-α-lapachone and (4)S) 4, 9-dihydroxyl-αLapachone, wherein (4R) 4, 9-dihydroxyl-α-lapachone and (4)S) 4, 9-dihydroxyl-αThe mass ratio of the lapachone is 50-60:40-50.
Further preferred is (4)R) 4, 9-dihydroxyl-α-lapachone and (4)S) 4, 9-dihydroxyl-αThe mass ratio of lapachone is 60:40.
Chiral IA high performance liquid chromatography conditions: chiral semi-preparative column CHIRALPAK ® IA column 10 mm X250 mm,5 μm; 95% of normal hexane is taken as a mobile phase A, ethanol is taken as a mobile phase B, and A: b=50: 50, performing isocratic elution according to the proportion; the sample injection amount is 10 mu L; the flow rate is 2.0 mL/min; the detection wavelength was 220 nm.
The 4, 9-dihydroxyl-containing material prepared by the inventionαThe beneficial effects of lapachone are:
the above 4, 9-dihydroxyl-containing material separated and purified from Sorbus bungeanaαThe preparation process of the lapachone takes the golden-silk catalpa heart wood as a raw material, has wide sources, simple and safe preparation process and high yield, and solves the problems of the prior artThere is no clear method for extracting 4, 9-dihydroxyl-containing material from catalpa bungeiαLapachone and chiral compounds (4)R) 4, 9-dihydroxyl-α-lapachone and (4)S) 4, 9-dihydroxyl-αLapachone, limiting the technical problems of utilization and research of catalpa bungei. And 4, 9-dihydroxyl-containing articles of the inventionαLapachone and chiral compounds (4)R) 4, 9-dihydroxyl-α-lapachone and (4)S) 4, 9-dihydroxyl-αThe lapachone has better anti-tumor activity, can be used as a medicine source medicine for preparing anti-tumor medicines, and has good medicinal prospect.
Drawings
Fig. 1: the 4, 9-dihydroxyl-coated substrates prepared in example 1 of the present inventionαLapaquinone sample, and 4, 9-dihydroxyl-αLapachone, (4)R) 4, 9-dihydroxyl-α-lapachone and (4)S) 4, 9-dihydroxyl-α-lapachone chemical formula.
Fig. 2: 4, 9-dihydroxyl- & ltCHEM & gt in example 1 of the inventionα-lapachone AQ-C18 high performance liquid chromatography purity profile.
Fig. 3: 4, 9-dihydroxyl- & ltCHEM & gt in example 1 of the inventionα-lapachone chiral IA high performance liquid chromatography optical purity profile.
Fig. 4: in example 4 of the present invention, (4)R) 4, 9-dihydroxyl-α-lapachone and (4)S) 4, 9-dihydroxyl-α-lapachone chiral IA high performance liquid chromatography optical purity profile.
Detailed Description
The present invention is further illustrated by the following examples, but the present invention is not limited to these examples. The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and starting materials, unless otherwise specified, may be obtained from commercial sources and/or prepared according to known methods.
Example 1
Pulverizing Aronia melanocarpa, soaking the powder in 5 times of dichloromethane solvent at normal temperature for 72 h, filtering to obtain extractive solution, and distilling under reduced pressure to obtain Aronia melanocarpa extract; dissolving the extract 1.3 and g with 1.5 and mL dichloromethane, and mixing with 3 times of 200-300 mesh silica gel for use; root of Chinese characterMixing 200-300 mesh silica gel with petroleum ether, and loading into chromatographic column (column size: 30×500 mm, column height: 200 mm); mixing the stirred sample with petroleum ether with equal volume, loading the sample by a wet method, and using petroleum ether after loading: ethyl acetate = 10:1 (v/v) Eluting, collecting with 10 mL penicillin bottle, tracking and detecting by TLC thin layer chromatography, and detecting according to R f Segmenting with color, combining R f The fraction with the value of 0.16 and the same color is concentrated under reduced pressure, dried and weighed to obtain red crystals 0.4126 g, the yield of which is 31.74 percent, and the fraction is detected to be 4, 9-dihydroxyl-by AQ-C18 high performance liquid chromatographyαLapachone with a purity of 99% or more.
AQ-C18 high performance liquid chromatography conditions: the chromatographic column is AQ-C18 column 4.6X1250 mm,5 μm; the mobile phase is methanol (A phase) -water (B phase) (gradient: 0-20 min, 10-100% methanol, 20-30 min,100% methanol, 30-35 min, 100-10% methanol); the flow rate is 1.0 mL/min; the sample injection amount is 10 mu L; the detection wavelength is 254 nm; the column temperature was 30 ℃.
Example 2
Pulverizing Aronia melanocarpa, soaking the powder in 5 times of acetone solvent at normal temperature for 70 h, filtering to obtain extractive solution, and distilling under reduced pressure to obtain Aronia melanocarpa extract; dissolving the extract 1.6 and g with 2 mL acetone, and mixing with 200-300 mesh silica gel 3 times of the extract; mixing 200-300 mesh silica gel with petroleum ether according to the mass amount after sample mixing, and loading into chromatographic column (column size: 30×500 mm, column height: 250 mm) by wet method; mixing the stirred sample with petroleum ether with equal volume, loading the sample by a wet method, and using petroleum ether after loading: ethyl acetate = 10:1 (v/v) Eluting, collecting with 10 mL penicillin bottle, tracking and detecting by TLC thin layer chromatography, and detecting according to R f Segmenting with color, combining R f The fraction with the value of 0.16 and the same color is concentrated under reduced pressure, dried and weighed to obtain red crystals 0.1834 g, the yield of which is 11.46 percent, and the fraction is detected to be 4, 9-dihydroxyl-by AQ-C18 high performance liquid chromatographyαLapachone with a purity of 99% or more.
AQ-C18 high performance liquid chromatography conditions: the chromatographic column is AQ-C18 column 4.6X1250 mm,5 μm; the mobile phase is methanol (A phase) -water (B phase) (gradient: 0-20 min, 10-100% methanol, 20-30 min,100% methanol, 30-35 min, 100-10% methanol); the flow rate is 1.0 mL/min; the sample injection amount is 10 mu L; the detection wavelength is 254 nm; the column temperature was 30 ℃.
Example 3
Pulverizing Aronia melanocarpa, soaking the powder in ethanol solvent 5 times of the powder at normal temperature for 74-h, filtering to obtain extractive solution, and distilling under reduced pressure to obtain Aronia melanocarpa extract; dissolving the extract 10.8 and g with 6 mL ethanol, and mixing with 3 times of 200-300 mesh silica gel; according to the mass quantity of 200-300 meshes of silica gel after sample mixing, loading the silica gel into a chromatographic column (column size: 90X 500 mm, column loading height: 230 mm) by a dry method; after dry loading of the stirred sample, petroleum ether is used for: ethyl acetate = 10:1 (v/v) Eluting, collecting with 500 mL glass bottle, tracking and detecting by TLC thin layer chromatography, and detecting according to R f Segmenting with color, combining R f The fraction with the value of 0.16 and the same color is decompressed, concentrated, dried and weighed to obtain red crystal 1.0784 g, the yield of which is 9.99 percent, and the fraction is detected to be 4, 9-dihydroxyl-percent by AQ-C18 high performance liquid chromatographyαLapachone with a purity of 99% or more.
AQ-C18 high performance liquid chromatography conditions: the chromatographic column is AQ-C18 column 4.6X1250 mm,5 μm; the mobile phase is methanol (A phase) -water (B phase) (gradient: 0-20 min, 10-100% methanol, 20-30 min,100% methanol, 30-35 min, 100-10% methanol); the flow rate is 1.0 mL/min; the sample injection amount is 10 mu L; the detection wavelength is 254 nm; the column temperature was 30 ℃.
Example 4
The 4, 9-dihydroxyl-containing material prepared in example 1 was takenαLapachone 30, mg, completely dissolved with 5 mL chromatographic ethanol, prepared by preparative HPLC on chiral semi-preparative column (4R) 4, 9-dihydroxyl-α-lapachone and (4)S) 4, 9-dihydroxyl-αLapachone. Obtain (4)R) 4, 9-dihydroxyl-αLapachone 14.3 mg and (4)S) 4, 9-dihydroxyl-αThe purity of the lapachone 9.7 and mg is over 99 percent by chiral IA high performance liquid chromatography detection.
Chiral IA highConditions for HPLC analysis: chiral semi-preparative column CHIRALPAK ® IA column 10 mm X250 mm,5 μm; 95% of normal hexane is taken as a mobile phase A, ethanol is taken as a mobile phase B, and A: b=50: 50, performing isocratic elution according to the proportion; the sample injection amount is 10 mu L; the flow rate is 2.0 mL/min; the detection wavelength was 220 nm.
In the above embodiments, the present invention is merely illustrative of the present invention but not limited thereto, and it should be understood that various modifications or alternatives may be made without departing from the scope of the present invention.
Claims (7)
1. The preparation method of the 4, 9-dihydroxyl-alpha-lapachone from the catalpa bungei is characterized by comprising the following steps of:
(1) Pulverizing Aronia melanocarpa, cold soaking in dichloromethane solvent, and distilling under reduced pressure to obtain extract;
(2) Loading the golden silk catalpa extract obtained in the step (1) to a silica gel chromatographic column, and eluting by a mixture of petroleum ether and ethyl acetate to obtain a crystal product, namely 4, 9-dihydroxyl-alpha-lapachone.
2. The method for preparing 4, 9-dihydroxy-alpha-lapachone from catalpa bungei according to claim 1, wherein the soaking temperature in the step (1) is normal temperature, and the soaking time is 70-74 h.
3. The method for preparing 4, 9-dihydroxy-alpha-lapachone from catalpa bungei according to claim 1, wherein the volume ratio of petroleum ether to ethyl acetate in the step (2) is 10-15:1.
4. the method for preparing 4, 9-dihydroxyl-alpha-lapachone from catalpa bungei according to claim 1, wherein the elution process in the step (2) is segmented according to the Rf value and the color, the parts with the same Rf value and the same color are combined, concentrated under reduced pressure, and dried to obtain red crystals, namely 4, 9-dihydroxyl-alpha-lapachone.
5. The method for preparing 4, 9-dihydroxy-alpha-lapachone from catalpa bungei according to claim 4, wherein the Rf value is 0.16, and the developing agent is petroleum ether: ethyl acetate mixed solvent (v/v, 10:1).
6. The 4, 9-dihydroxy- α -lapachone prepared by the method according to any one of claims 1 to 5, wherein the 4, 9-dihydroxy- α -lapachone comprises (4R) -4, 9-dihydroxy- α -lapachone and (4S) -4, 9-dihydroxy- α -lapachone, and wherein the mass ratio of (4R) -4, 9-dihydroxy- α -lapachone to (4S) -4, 9-dihydroxy- α -lapachone is 50-60:40-50.
7. The 4, 9-dihydroxy- α -lapachone according to claim 6, wherein the mass ratio of (4R) -4, 9-dihydroxy- α -lapachone to (4S) -4, 9-dihydroxy- α -lapachone is 60:40.
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| CN116410173A (en) * | 2023-02-28 | 2023-07-11 | 兰州大学 | Pyran naphthoquinone compound, preparation method, composition and application thereof |
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| CN116410173A (en) * | 2023-02-28 | 2023-07-11 | 兰州大学 | Pyran naphthoquinone compound, preparation method, composition and application thereof |
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