阅读说明：本技术 具有抗癌活性的6-溴代下箴刺桐碱及其制备方法和应用 (6-bromohypaphorine with anticancer activity and preparation method and application thereof ) 是由 戴雨霖 郑飞 越皓 于 2020-04-01 设计创作，主要内容包括：本发明提供一种具有抗癌活性的6-溴代下箴刺桐碱及其制备方法和应用,属于化合物制备方法领域。该6-溴代下箴刺桐碱的分子式是C<Sub>14</Sub>H<Sub>17</Sub>BrN<Sub>2</Sub>O<Sub>2</Sub>,化学式如式1所示。本发明还提供一种具有抗癌活性的6-溴代下箴刺桐碱的制备方法。本发明还提供上述6-溴代下箴刺桐碱在制备抗癌药物方面的应用。本发明的6-溴代下箴刺桐碱是首次从红仿刺参中提取出来的,该提取方法简单、产率大、纯度高,易于产业化,同时得到的化合物可以应用在制备抗癌药物上,尤其对肺癌效果显著。<Image he="273" wi="700" file="DDA0002434770320000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>(The invention provides 6-bromohypaphorine with anticancer activity and a preparation method and application thereof, belonging to the field of compound preparation methods. The molecular formula of the 6-bromohypaphorine is C 14 H 17 BrN 2 O 2 The chemical formula is shown as formula 1. The invention also provides a preparation method of 6-bromohypaphorine with anticancer activity. The invention also provides the application of the 6-bromohypaphorine in preparing anti-cancer drugs. The 6-bromohypaphorine is extracted from the red apostichopus japonicus for the first time, the extraction method is simple, the yield is high, the purity is high, the industrialization is easy, and the obtained compound can be applied to the preparation of anti-cancer drugs, and particularly has obvious effect on lung cancer.)

1. 6-bromohypaphorine with anticancer activity, characterized in that the molecular formula is C₁₄H₁₇BrN₂O₂The chemical formula is shown as formula 1:

2. the process for the preparation of 6-bromohypaphorine with anticancer activity according to claim 1, wherein the process comprises:

the method comprises the following steps: raw material collection and treatment

Cleaning red apostichopus japonicus with clear water, reserving internal organs, freeze-drying, crushing, and sampling to obtain a crushed sample;

step two: extraction separation

Heating and shaking the crushed sample obtained in the first step by adopting an ethanol solution for extraction, recovering an extract under reduced pressure, sequentially extracting by using n-hexane, chloroform, ethyl acetate and n-butyl alcohol, wherein the volume ratio of the sample solution to an extraction reagent is 1 (2-3), extracting for three times by using each extraction liquid, combining the extraction liquids, separating components obtained by n-butyl alcohol extraction in a normal-phase preparation thin-layer plate, separating the components by using chloroform, methanol and water as mobile phases, and collecting the components with a component ratio shift value of 0.06-0.10;

step three: enriching and purifying

The components separated by the thin layer plate are enriched and purified 6-bromohypaphorine in a reversed-phase preparation liquid phase, the mobile phase is ultrapure water and acetonitrile solution, the gradient of the mobile phase is 0-30min, the volume ratio of water to acetonitrile is 50% and 50%, 30-40min, the volume ratio of water to acetonitrile is 90% and 10%, the size of a preparation chromatographic column of ODS is 10mm inner diameter, 250mm length and 5um particle size, the flow rate of the preparation liquid phase is 2m L/min, and the apocrine flows out under 6-bromo in 25.63 min under an ultraviolet lamp 254 nm.

3. The method of claim 2, wherein the first step of pulverizing the precursor is followed by sieving through a sieve of four to six meshes to obtain a pulverized sample.

4. The process of claim 2, wherein the concentration of the diethylene alcohol solution is 70-95%.

5. The method for preparing 6-bromohypaphorine with anticancer activity according to claim 2, wherein the heating temperature in step two is 40-50 ℃ and the extraction time is 8-12 h.

6. The method of claim 2, wherein the volume ratio of chloroform, methanol and water is 10:1 (0.1-0.3).

7. Use of 6-bromohypaphorine according to claim 1 for the preparation of an anti-cancer medicament.

8. Use of 6-bromohypaphorine according to claim 7, wherein the anti-cancer agent is a lung cancer agent.

Technical Field

The invention belongs to the field of compound preparation methods, and particularly relates to 6-bromohypaphorine with anticancer activity, a preparation method and application thereof.

Background

Marine organisms use their own secreted metabolites as a tool to resist enemy attack, and thus some marine secondary metabolites have a good anticancer effect. Apostichopus japonicus (Apostichopus japonicus) belongs to invertebrates and is mainly distributed in east Asia areas, especially in the yellow sea area of China. The traditional Chinese medicine considers that the apostichopus japonicus is used as a medicine for the whole body and is a superior nourishing material. Modern researches consider that the apostichopus japonicus has the effects of resisting cancers, fungi, inflammation, intelligence and the like.

Recent research at home and abroad shows that the compound of the hundreds of species is extracted and separated from the sea cucumber, wherein the content of saponins, proteins and polysaccharides is high and the activity is obvious.

Disclosure of Invention

The invention aims to provide 6-bromohypaphorine with anticancer activity, a preparation method and application thereof.

The invention firstly provides 6-bromohypaphorine with anticancer activity, the molecular formula is C₁₄H₁₇BrN₂O₂The chemical formula is shown as formula 1:

the invention also provides a preparation method of 6-bromohypaphorine with anticancer activity, which comprises the following steps:

the method comprises the following steps: raw material collection and treatment

Cleaning red apostichopus japonicus with clear water, reserving internal organs, freeze-drying, crushing, and sampling to obtain a crushed sample;

step two: extraction separation

Heating and shaking the crushed sample obtained in the first step by adopting an ethanol solution for extraction, recovering an extract under reduced pressure, sequentially extracting by using n-hexane, chloroform, ethyl acetate and n-butyl alcohol, wherein the volume ratio of the sample solution to an extraction reagent is 1 (2-3), extracting for three times by using each extraction liquid, combining the extraction liquids, separating components obtained by n-butyl alcohol extraction in a normal-phase preparation thin-layer plate, separating the components by using chloroform, methanol and water as mobile phases, and collecting the components with a component ratio shift value of 0.06-0.10;

step three: enriching and purifying

The components separated by the thin layer plate are enriched and purified 6-bromohypaphorine in a reversed-phase preparation liquid phase, the mobile phase is ultrapure water and acetonitrile solution, the gradient of the mobile phase is 0-30min, the volume ratio of water to acetonitrile is 50% and 50%, 30-40min, the volume ratio of water to acetonitrile is 90% and 10%, the size of a preparation chromatographic column of ODS is 10mm inner diameter, 250mm length and 5um particle size, the flow rate of the preparation liquid phase is 2m L/min, and the apocrine flows out under 6-bromo in 25.63 min under an ultraviolet lamp 254 nm.

Preferably, the crushed sample is obtained by sieving the crushed sample in the first step through a sieve from four to six.

Preferably, the concentration of the ethanol solution in the step is 70-95%.

Preferably, the heating temperature in the second step is 40-50 ℃, and the extraction time is 8-12 h.

Preferably, the volume ratio of the chloroform to the methanol to the water in the second step is 10:1 (0.1-0.3).

The invention also provides the application of the 6-bromohypaphorine in preparing anti-cancer drugs.

Preferably, the anti-cancer drug is a lung cancer drug.

The invention has the advantages of

The invention provides a 6-bromo hypaphorine with anticancer activity and a preparation method and application thereof, the 6-bromo hypaphorine is firstly extracted from red apostichopus japonicus, the extraction method is simple, the yield is high, the purity is high, the industrialization is easy, and the obtained compound can be applied to the preparation of anticancer drugs, and especially has obvious effect on lung cancer.

Drawings

FIG. 1 shows a COSY spectrum of 6-bromohypaphorine prepared in example 1 of the present invention;

FIG. 2 shows a HMBC chromatogram of 6-bromohypaphorine prepared according to example 1 of the present invention;

FIG. 3 shows the HSQC spectrum of 6-bromohypaphorine prepared according to example 1 of the present invention;

FIG. 4 is a graph showing the cell viability of 6-bromohypaphorine on normal cells and five kinds of cancer cells in example 3 of the present invention;

FIG. 5 is a fluorescent micrograph of acridine orange-ethidium bromide and Hoechst 33342 of example 3 of the present invention.

FIG. 6 is an electrophoretogram of example 3 of the present invention.

Detailed Description

The invention firstly provides 6-bromohypaphorine with anticancer activity, the molecular formula is C₁₄H₁₇BrN₂O₂The powder is white amorphous powder, and the chemical formula is shown as formula 1:

the invention also provides a preparation method of 6-bromohypaphorine with anticancer activity, which comprises the following steps:

the method comprises the following steps: raw material collection and treatment

Cleaning red apostichopus japonicus with clear water, keeping viscera, freeze-drying until the powder is completely dried, and crushing, wherein the sample is preferably sieved by a sieve from four to six to obtain a crushed sample; the red Apostichopus japonicus is preferably fresh red Apostichopus japonicus (Apostichopus japonicus) collected in spring in the yellow sea area;

step two: extraction separation

Heating the crushed sample obtained in the first step by adopting an ethanol solution, shaking and extracting by a shaking table, wherein the concentration of the ethanol solution is preferably 70-95% (v/v), the heating temperature is preferably 40-50 ℃, the extraction time is preferably 8-12h, recovering an extract under reduced pressure, sequentially extracting by using n-hexane, chloroform, ethyl acetate and n-butyl alcohol, the volume ratio of the sample solution to an extraction reagent is 1 (2-3), extracting each extraction liquid for three times, combining the extraction liquids, separating components obtained by n-butyl alcohol extraction in a normal-phase preparation thin-layer plate, separating a mobile phase by using chloroform, methanol and water, wherein the volume ratio of the chloroform to the methanol to the water is preferably 10:1 (0.1-0.3), and the ratio shift value of the collected components is 0.06-0.10;

step three: enriching and purifying

The components separated by the thin layer plate are enriched and purified 6-bromohypaphorine in a reversed-phase preparation liquid phase, the mobile phase is ultrapure water and acetonitrile solution, the gradient of the mobile phase is 0-30min, the volume ratio of water to acetonitrile is 50% and 50%, 30-40min, the volume ratio of water to acetonitrile is 90% and 10%, the size of a preparation chromatographic column of ODS is 10mm inner diameter, 250mm length and 5um particle size, the flow rate of the preparation liquid phase is 2m L/min, and the apocrine flows out under 6-bromo in 25.63 min under an ultraviolet lamp 254 nm.

The invention also provides the application of the 6-bromohypaphorine in preparing anti-cancer drugs. The anti-cancer drug is preferably a lung cancer drug.

The present invention is described in further detail below with reference to specific examples, in which the starting materials are all commercially available.