CN101422466B - Use of cucurbitacin in preparing medicine for treating cancer - Google Patents

Abstract

The invention relates to the new application of cucurbitacin in preparing a medicament for curing cancers. The invention relates to the application of the cucurbitacin in preparing a medicametn for curing liver cancer. Test proves that the cucurbitacin has remarkable inhibition or killing effects on various cancers such as liver cancer, lung cancer, gastric cancer, laryngocarcinoma, leukemia, and the like. The action intensity of the cucurbitacin is relevant to a preparation formulation, a medication approach and dosage.

Description

The application of cucurbitacin in the medicine of preparation treatment cancer

The application is 200610010906.7 divides an application, and the applying date of original application is on May 19th, 2006, and application number is 200610010906.7, and invention and created name is: the preparation method of cucurbitacin and the application in the medicine of preparation treatment cancer

Technical field

The present invention relates to a kind of preparation method of medicinal component cucurbitacin, and the new application in the medicine of preparation treatment cancer.

Background technology

Hemsleyadin is a kind of mixture, is made up of cucurbitacin and dihydrocucurbitacin F.The earliest by people such as Chen Weixin from the root portion of garden fruit Radix Hemsleyae Macrospermae plant (Hemsleya amalils Diels) from obtaining (chemical journal, 1975,33 (1), 5), cucurbitacin is accredited as 19-mistake carbon--9 Beta-methyls-10 à-3 à-hydrogen-2 β, 3 à, 16 à, 20,25-penta hydroxy group Δ 5 lanostene-11,20-diketone-25-acetas (IIa) is the two hydrogen of the 23.24-cc of cucurbitacin F (Ic)-25-acetas, and the second element is accredited as 23 of cucurbitacin F (Ic), the two hydrogen compounds of 24-, be two kinds of new chemical compounds, and development hemsleyadin tablet, recorded medicine drug standard in 1974 in Yunnan Province, be used for the treatment of bacillary dysentery enteritis, tracheitis, the acute tonsillitis.

In order to seek the plant resources that extracts hemsleyadin, some scholars have successively carried out the research of resource chemistry to this platymiscium, successively from Radix Hemsleyae Macrospermae (Hemsleya maerosperma c.y.wu), dolichocarpel hemsleya root (H, dolichocarp W.J.cheng) high eyebrow Radix Hemsleyae Macrospermae (H, emiensil L.T.Sent) isolates cucurbitacin, the second element, the extracting method of its extracting method and Chen Shi roughly the same, all use the ethanol extraction crude drug, extract extractum and in water, use ethyl acetate extraction, silicagel column on the abstract, use chloroform: acetone: the mixed solvent eluting of three kinds of different proportions of ethyl acetate, eluent reclaim crude product, the reuse ethyl alcohol recrystallization can obtain the first element respectively, second element (list of references: Yunnan animals and plants researchs such as Rui-Lin-Nie etal.planta Medica1984:322: Shi Yaqin.1990。12 (4), 460; Chinese herbal medicine 1995,26 (12), 619).

Chinese patent application CN01102620.0 discloses a kind of " nano medicine ' Xuedansu ' and preparation method thereof ", be to be raw material with the nanometer hemsleyadin, preparation in proportion, make new pharmaceutical preparation, its fineness of the particles reaches 1200-1500 orders, particle diameter is 0.1-200nm, and wherein most particle diameters are less than 100nm, and has new rerum natura.Adopt steps such as microwave extracting, concentrating under reduced pressure, supersonic jet technology spray drying to make.

Chinese patent application CN03159197.3 discloses a kind of " drop pills of hemsleyadin and preparation method thereof ", it is characterized in that: the hemsleyadin fine powder of 1 weight portion through micronizing is added in 1～10 weight portion molten matrix, abundant mixing, dropping preparation method is condensed into ball in coolant, remove coolant, drying, promptly.That Chinese patent application CN02134164.8 discloses is a kind of " hemsleyadin analog derivative and preparation method thereof and as the application of antibacterial ", a kind of hemsleyadin derivant unlike the prior art is provided, its preparation method is the antibacterial of effective ingredient with this chemical compound.

At present, do not find the report that cucurbitacin is used as yet in anticancer medicine.

Summary of the invention

The object of the present invention is to provide a kind of preparation method of cucurbitacin.

Another object of the present invention provides the application of cucurbitacin in the medicine of preparation treatment cancer.

The preparation method of cucurbitacin of the present invention is made up of following steps:

One, getting Radix Hemsleyae Macrospermae platymiscium tuber, pulverize, is solvent with the acetone of 18-26 times of weight, adopts the percolation extraction, and extracting solution reclaims in 60-75 ℃ of water-baths, promptly gets extractum;

Two, the extractum that proposes is admixed the silica gel of 1-2 times of weight, oven dry, last silicagel column, use chloroform: methanol carries out gradient elution, and wherein the methanol volumetric concentration is 2-10%, reclaims eluent, gets crude product;

Three, with methanol or ethanol to crude product heating for dissolving in water-bath, filter, put room temperature, recrystallization must be insulted column crystallization, promptly purity is the cucurbitacin more than 98%.

Radix Hemsleyae Macrospermae platymiscium described in the step 1 is Radix Hemsleyae Macrospermae, dolichocarpel hemsleya root, high eyebrow Radix Hemsleyae Macrospermae etc.First in the step 2 with pure chloroform eluting impurity, reuse chloroform: methanol gradient elution; The gained cucurbitacin more than 98%, fusing point 226-228 ℃, through ultraviolet spectra, infrared spectrum, nuclear magnetic resonance, NMR, mass spectrum four analysis of spectrums, is confirmed as cucurbitacin through high performance liquid chromatogram (HPLC) purity assay in the step 3.Its structural formula is:

The present invention relates to the application of cucurbitacin in the medicine of preparation treatment hepatocarcinoma.

The present invention relates to the application of cucurbitacin in preparation treatment lung cancer drugs.

The present invention relates to the application of cucurbitacin in the medicine of preparation treatment gastric cancer.

The present invention relates to the application of cucurbitacin in the medicine of preparation treatment laryngeal carcinoma.

The present invention relates to the application of cucurbitacin in the leukemic medicine of preparation treatment.

The pharmacodynamics test data:

Cucurbitacin monomer composition is through the anticancer test in pharmacology inside and outside, affirmation this product is external to have inhibition or killing action to human body solid carcinoma cell, Ic50 (μ g/m1) to the growth of human body lung cell A549 is 1.50, basic identical with the Ic501.60 of cisplatin, test of mice vivo antitumor and preparation type, and route of administration is relevant, by contrast, oral administration is poorer slightly than drug administration by injection, and its Emulsion intravenous injection reaches as high as 59% to the pulmonary carcinoma suppression ratio.

Anticancer test:

1, body outer screening test adopts the active improvement mtt assay of cancer-resisting substance to carry out.

JEG-3:

The HL60 human leukemia cell line

The A549 human lung carcinoma cell line

The SGC-7901 human stomach cancer cell line

The Be1-7402 human hepatoma cell strain

The SMMC-7721 human hepatoma cell strain

The strain of HEP-2 people's laryngeal cancer cell

The GLC-15 human lung carcinoma cell line

Experimental result:

IC50 (μ g/ml) to 7 strain growth of human tumor cells

From last table as seen, under this experiment condition, cucurbitacin demonstrates comparatively significantly killing action to human body tumour cell, but the tumor cell of different tissue sources is had than evident difference, and the prompting cucurbitacin has relative selectivity to different tumor cells.

2, in vivo test

Adopt kunming mice to inoculate H22 hepatocarcinoma and Mice Bearing Lewis Lung Cancer respectively, adopt three kinds of preparations, suspensoid, Emulsion, three kinds of route of administration of lyophilized formulations, irritate stomach, lumbar injection, intravenous injection, dosage divides high, medium and low three therapeutic doses, compare with positive control, calculate the antitumor suppression ratio, experimental result is each table of face as follows:

Table 1 cucurbitacin suspensoid gastric infusion is to the clinical trial of mice H22 hepatocarcinoma

Compare * * * P＜0.01 with negative control group.This experiment has significant difference.

Table 2 cucurbitacin suspensoid intraperitoneal administration connects swollen clinical trial to mice H22 hepatocarcinoma

Sample

Dosage

Dosage regimen

Number of animals (only)

The weight of animals (g)

Tumor heavy (g)

Suppression ratio

mg/kg/d

igx10qd

Beginning/end

Beginning/end

X±SD

Cucurbitacin

90

igx10qd

10/9

21.1/27.6

1.31±0.12

57.05

Cucurbitacin

60

igx10qd

10/10

21.0/24.6

1.49±0.12

51.05

Cucurbitacin

30

igx10qd

10/10

20.8/25.4

1.95±0.12

36.07

Positive control CTX

30

ipx7qd

10/10

21.2/23.1

0.33±0.13

89.18

Negative control

Coordinative solvent

igx10qd

20/20

20.8/25.9

3.05±0.28

Compare * * * P＜0.01 with negative control group.

Table 3 cucurbitacin Emulsion is to mice H22 hepatocarcinoma intravenously administrable test curative effect table

Sample

Dosage

Dosage regimen

Number of animals (only)

The weight of animals (g)

Tumor heavy (g)

Suppression ratio

mg/kg/d

ivx10qd

Beginning/end

Beginning/end

X±SD

Cucurbitacin

15

ivx10qd

10/10

19.5/25.3

1.48±0.11

55.02

Cucurbitacin

10

ivx10qd

10/10

19.3/25.9

1.76±0.08

44.12

Cucurbitacin

5

ivx10qd

10/10

19.4/25.1

2.13±0.20

32.38

Positive control CTX

30

ivx7qd

10/10

19.6/22.9

0.31±0.13

90.16

Negative control

Coordinative solvent

ivx10qd

10/10

19.4/26.0

3.15±0.44

Compare * * * P＜0.01 with negative control group.

Table 4 cucurbitacin Emulsion is to Mice Bearing Lewis Lung Cancer intravenously administrable test curative effect table

Sample

Dosage

Dosage regimen

Number of animals (only)

The weight of animals (g)

Tumor heavy (g)

Suppression ratio

mg/kg/d

ivx10qd

Beginning/end

Beginning/end

X±SD

Cucurbitacin

15

ivx10qd

10/10

19.5/22.1

1.16±0.16

59.30

Cucurbitacin

10

ivx10qd

10/10

19.1/22.9

1.50±0.14

47.37

Cucurbitacin

5

ivx10qd

10/10

19.3/22.4

1.69±0.11

40.76

Positive control CTX

30

ivx10qd

10/10

18.7/18.1

0.362±0.03

87.76

Negative control

Coordinative solvent

ivx10qd

20/20

18.9/23.0

2.85±0.23

Compare * * * P＜0.01 with negative control group.

Table 5 cucurbitacin lyophilized preparation is to mice H22 hepatocarcinoma intravenously administrable test curative effect table

Sample

Dosage

Dosage regimen

Number of animals (only)

The weight of animals (g)

Tumor heavy (g)

Suppression ratio

mg/kg/d

ivx10qd

Beginning/end

Beginning/end

X±SD

Cucurbitacin

15

ivx10qd

10/10

20.4/26.7

1.61±0.18

46.15

Cucurbitacin

10

ivx10qd

10/10

20.7/25.9

1.82±0.19

39.13

Cucurbitacin

5

ivx10qd

10/10

20.5/26.8

2.06±0.18

31.1

Positive control CTX

30

Ipx7qd

10/10

20.9/24.1

0.402±0.14

86.56

Negative control

Coordinative solvent

ivx10qd

20/20

20.3/27.3

2.99±0.30

Table 6 cucurbitacin lyophilized preparation is to the prerun of Mice Bearing Lewis Lung Cancer (subcutaneous vaccination) curative effect

Sample

Dosage

Dosage regimen

Number of animals (only)

The weight of animals (g)

Tumor heavy (g)

Suppression ratio

mg/kg/d

ivx10qd

Beginning/end

Beginning/end

X±SD

Cucurbitacin

15

ivx10qd

10/10

18.0/20.7

1.20±0.20

51.20

Cucurbitacin

10

ivx10qd

10/10

17.7/21.1

1.34±0.18

45.31

Cucurbitacin

5

ivx10qd

10/10

18.1/20.5

1.56±0.14

36.33

DDP

7

Ipx2, the 1st, 3 day

10/10

18.4/18.1

0.172±0.06

92.98

Negative control

NS

ivx10qd

20/20

17.9/22.4

2.45±0.22

* * P value＜0.01 is compared with negative control group.

Conclusion: to various cancers, comprise hepatocarcinoma by the above-mentioned cucurbitacin that experiment showed,, pulmonary carcinoma, gastric cancer, laryngeal carcinoma, leukemia etc. have significant inhibition or killing action, and its action intensity is relevant with dosage form, route of administration and dosage.

The specific embodiment

Embodiment 1: get area, Qiaojia County, Yunnan Radix Hemsleyae Macrospermae 5kg, be ground into middle powder, Sheng is infiltrated in the fluid capacitance device, at 15-25 ℃, add 24 times of amount acetone and soak after 48 hours, carry out percolation, reclaim percolate by the Chinese Pharmacopoeia percolation, extract is admixed the silica gel of equivalent, 50-70 ℃ of drying, last silicagel column, use chloroform: methanol is that eluent carries out gradient elution, be collected in the eluent of 3-5% methanol concentration, can obtain crude product after the recovery, with methanol recrystallization repeatedly, Ling Zhujing, 226-228 ℃ of fusing points, HPLC purity assay 98.98%.See Fig. 1,, confirm as cucurbitacin through ultraviolet, infrared spectrum, nuclear magnetic resonance, NMR, mass spectrum four analysis of spectrums.

With above-mentioned cucurbitacin is monomer component, makes dosage forms such as suspensoid, Emulsion, lyophilized formulations, nanometer lyophilized preparation, tablet, pill, drop pill, capsule, controlled release agent, microcapsule, tablet, liposome according to common process.Be used for hepatocarcinoma, pulmonary carcinoma, gastric cancer, laryngeal carcinoma and leukemic treatment.

Claims (1)

1. the application of cucurbitacin in the medicine of preparation treatment hepatocarcinoma.