Preparation method for marine microorganism natural and polymorphic arsenic compound extract for inhibiting tumors and application thereof

Abstract

The invention discloses a preparation method for marine microorganism natural and polymorphic arsenic compound extract for inhibiting tumors, comprising the following steps of: (1) preparing fine arsenic mineral powder for extracting; (2) metabolizing the ultrafine arsenic mineral powder by marine microorganisms to obtain arsenic extract. The invention also discloses the application of the arsenic compound extract in antitumor drugs. The active components of the antitumor drugs of the arsenic compound extract are specific. Compared with similar and first-line anticancer compound arsenious acids, the arsenic compound extract has the characteristics of enhanced effect and reduced toxicity.

Description

technical field

[0001] The invention relates to a preparation method and application of marine microorganisms to extract natural, polymorphic and tumor-inhibiting arsenic-containing compound extracts. Background technique

[0002] Since the advent of pure arsenic trioxide injection developed in my country, it has brought good news to patients due to its good anti-tumor effect, prolonging life and also bringing good social benefits. However, the currently used arsenic trioxide injection is generally made of high-purity chemicals, which still has the disadvantages of high price, increased liver and kidney toxicity in clinical use, and difficulty in popularization. Therefore, finding more natural arsenic-containing drugs and developing more natural, low-toxicity and high-efficiency arsenic-containing compounds as supplements or substitutes for the existing first-line anticancer arsenic arsenic trioxide will be an urgent requirement for modern innovative Chinese medicine. . Re...

Examples

Embodiment 1

[0017] The preparation method of a marine microorganism extracting natural, polymorphic and tumor-inhibiting arsenic compound extract of this embodiment includes the following steps: 1) preparing arsenic ore fine powder for extraction; 2) marine microorganisms metabolizing arsenic ore super Micro powder fine powder. The arsenopyrite ore described is arsenopyrite (FeAsS), arsenopyrite (FeAsS), orthopedic arsenite (FeAsS), 2 A S3 ), nickel arsenic ore (NiAS 2 ), red arsenic nickel ore (NiAS), arsenic cobalt ore (CoAsS 2 ), arsenite (Cu4As12S13), arsenite (Cu 3 AsS 4 ), arsenic platinum ore (PtAs 2 ), cobalt ore [C(OFe)AsS], realgarite (As 2 o 3 , cockscomb stone), orpiment (As 2 S 3 ) of one or more natural minerals. Of course, the above 12 kinds of arsenic-containing minerals are relatively common, but the present invention is not limited to these 12 kinds, and other arsenic-containing minerals are also applicable. The steps for preparing the fine powder of arsenic o...

Embodiment 2

[0023] Animal acute toxicity test of the arsenic-containing compound extract prepared by the present invention: the present embodiment selects the arsenic-containing compound extract prepared by the preferred combination of Alcaligenes seawater, Corynebacterium blunt-toothed and Brevibacterium flavum in equal proportions in Example 1 .

[0024] 20 mice, half male and half male, one-time ig 0.2ml / 10g of the extract of the present invention (arsenic acid V, 289.3mg / mL; monomethylarsenic acid V, 168.3mg / mL, dimethylarsenic acid V, 69.7 mg / mL), and then observe the activity and death of the mice for 7 days. It was found that the 2 S 3 Compared with conventional coarse powder, the number of mouse deaths was reduced, as can be seen from the results in Table 3, (Table 3 shows the changes in the distribution of arsenic in the body of mice after oral administration of the extract prepared by the present invention and arsenous acid) As 2 S 3 LD of the extract 50 Value is significan...

Embodiment 3

[0027] This embodiment is a test for the determination of tumor inhibitory activity: the myeloma cell line U266 in the logarithmic growth phase is collected, and the cell density is adjusted to 1 × 10 in a 50ml cell culture flask. 6 cells / flask. Add an appropriate amount of the extract of the present invention or sodium arsenite injection, and adjust the final concentration of both to 2.5umol / L. After culturing for 24 h, the cells were collected and washed with cold phosphate buffered solution (PBS, containing 8.0 g of NaCl, 0.20 g of KCl, and NaCl per liter). 2 HPO 4 1.15 g, KH 2 PO 4 0.20 g, pH 7.4) and washed twice, add FITC-Annexin V and PI, mix gently, place in the dark at 4 °C for 30 min, add binding buffer (BindingBuffer), and detect with flow cytometry within 1 h Its early apoptosis rate and late apoptosis rate. The results are shown in Table 4. Table 4 is a comparison of the apoptosis rate of the myeloma cell U266 induced by the extract prepared by the present...

technical field

[0001] The invention relates to a preparation method and application of marine microorganisms to extract natural, polymorphic and tumor-inhibiting arsenic-containing compound extracts. Background technique

[0002] Since the advent of pure arsenic trioxide injection developed in my country, it has brought good news to patients due to its good anti-tumor effect, prolonging life and also bringing good social benefits. However, the currently used arsenic trioxide injection is generally made of high-purity chemicals, which still has the disadvantages of high price, increased liver and kidney toxicity in clinical use, and difficulty in popularization. Therefore, finding more natural arsenic-containing drugs and developing more natural, low-toxicity and high-efficiency arsenic-containing compounds as supplements or substitutes for the existing first-line anticancer arsenic arsenic trioxide will be an urgent requirement for modern innovative Chinese medicine. . Re...

Examples

Embodiment 1

[0017] The preparation method of a marine microorganism extracting natural, polymorphic and tumor-inhibiting arsenic compound extract of this embodiment includes the following steps: 1) preparing arsenic ore fine powder for extraction; 2) marine microorganisms metabolizing arsenic ore super Micro powder fine powder. The arsenopyrite ore described is arsenopyrite (FeAsS), arsenopyrite (FeAsS), orthopedic arsenite (FeAsS), 2 A S3 ), nickel arsenic ore (NiAS 2 ), red arsenic nickel ore (NiAS), arsenic cobalt ore (CoAsS 2 ), arsenite (Cu4As12S13), arsenite (Cu 3 AsS 4 ), arsenic platinum ore (PtAs 2 ), cobalt ore [C(OFe)AsS], realgarite (As 2 o 3 , cockscomb stone), orpiment (As 2 S 3 ) of one or more natural minerals. Of course, the above 12 kinds of arsenic-containing minerals are relatively common, but the present invention is not limited to these 12 kinds, and other arsenic-containing minerals are also applicable. The steps for preparing the fine powder of arsenic o...

Embodiment 2

[0023] Animal acute toxicity test of the arsenic-containing compound extract prepared by the present invention: the present embodiment selects the arsenic-containing compound extract prepared by the preferred combination of Alcaligenes seawater, Corynebacterium blunt-toothed and Brevibacterium flavum in equal proportions in Example 1 .

[0024] 20 mice, half male and half male, one-time ig 0.2ml / 10g of the extract of the present invention (arsenic acid V, 289.3mg / mL; monomethylarsenic acid V, 168.3mg / mL, dimethylarsenic acid V, 69.7 mg / mL), and then observe the activity and death of the mice for 7 days. It was found that the 2 S 3 Compared with conventional coarse powder, the number of mouse deaths was reduced, as can be seen from the results in Table 3, (Table 3 shows the changes in the distribution of arsenic in the body of mice after oral administration of the extract prepared by the present invention and arsenous acid) As 2 S 3 LD of the extract 50 Value is significan...

Embodiment 3

[0027] This embodiment is a test for the determination of tumor inhibitory activity: the myeloma cell line U266 in the logarithmic growth phase is collected, and the cell density is adjusted to 1 × 10 in a 50ml cell culture flask. 6 cells / flask. Add an appropriate amount of the extract of the present invention or sodium arsenite injection, and adjust the final concentration of both to 2.5umol / L. After culturing for 24 h, the cells were collected and washed with cold phosphate buffered solution (PBS, containing 8.0 g of NaCl, 0.20 g of KCl, and NaCl per liter). 2 HPO 4 1.15 g, KH 2 PO 4 0.20 g, pH 7.4) and washed twice, add FITC-Annexin V and PI, mix gently, place in the dark at 4 °C for 30 min, add binding buffer (BindingBuffer), and detect with flow cytometry within 1 h Its early apoptosis rate and late apoptosis rate. The results are shown in Table 4. Table 4 is a comparison of the apoptosis rate of the myeloma cell U266 induced by the extract prepared by the present...

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