WO2017107993A1 - Use of polyether compound - Google Patents

Abstract

A polyether compound shown in general formula 1, a use of the chelated form, hydrated form or pharmaceutically acceptable salt thereof in the preparation of a medicine, and a use of the medicine in the preparation of a medicine for treating cancer or preventing the recurrence of cancer. The polyether compound shown in formula 1 can inhibit the proliferation of malignant tumor cells and malignant tumor stem cells.

Description

Polyether compound use

Priority information

The present application claims priority to and the benefit of the patent application No. 201511000381.4, filed on Dec. 24, 2015, the disclosure of which is hereby incorporated by reference.

Technical field

The present invention relates to the field of medicinal chemistry and pharmacotherapeutics, and in particular to polyether compounds, processes for their preparation, pharmaceutical compositions containing such compounds, and uses.

Background technique

Cancer has become one of the most important diseases that endanger human health. At present, it usually includes surgical treatment, natural therapy, radiation therapy, chemotherapy, and Chinese medicine treatment. Chemotherapy refers to the purpose of killing or inhibiting the growth of tumor cells by using chemotherapeutic drugs. The chemotherapeutic drugs can be divided into alkylating agents, antimetabolites, anticancer antibiotics, plants, hormones and impurities, among which existing Although anticancer antibiotics can achieve certain effects, long-term use can easily lead to drug resistance of tumor cells, and often cause toxic side effects. Therefore, it is necessary to expand the diversity of such anticancer drugs. The present inventors have found that ether antibiotics have a good effect of inhibiting tumor growth.

Summary of the invention

The present invention aims to solve at least one of the technical problems existing in the prior art.

The present invention aims to solve at least one of the technical problems in the related art to some extent. To this end, it is an object of the present invention to provide a means for effectively treating or preventing cancer.

According to one aspect of the present invention, there is provided a use of a polyether compound of the formula I, a chelated form thereof, a hydrated form or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment Cancer and prevent cancer from recurring,

Wherein R1, R2, R3, R4, R5, R6, R7 and R8 are each independently hydrogen, -OH, -NH2, halogen, optionally substituted a C1-10 linear or branched saturated or unsaturated hydrocarbon group, an optionally substituted C1-10 linear or branched alkoxy group, an optionally substituted C1-10 acyl group, an optionally substituted C6-20 aryl group , optionally substituted heteroaryl,

or

Wherein R ₉ and R ₁₀ are each independently hydrogen, -OH, -NH ₂ , halogen, C _1-5 linear or branched saturated or unsaturated hydrocarbon group, carboxyl group, azide group, sulfhydryl group, amine group, a mercapto group, an amide group, sulfate group, rhamnose group, phenyl or benzyl, and optionally substituted with one or more substituents selected from _{halo, -OCH 3, -OH, -OCF 3} , -NH 2, phenyl Substituted by a group of a benzyl group, a heteroaryl group, and -NHR ^X and -NR ^X ₂ , wherein each R ^{X is} independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group, The heteroaryl group is a 3- to 6-membered monocyclic or bicyclic heteroaryl group, optionally containing 1 to 3 hetero atoms selected from N, O or S;

n is an integer of 2-7;

The halogen is -F, -Cl, -Br or -I.

In addition, it should be noted that the drug of the present invention not only inhibits the growth of tumor cells, but also inhibits the growth of cancer stem cells, thereby completely eliminating cancer cells, preventing cancer recurrence, and particularly preventing breast cancer recurrence.

According to an embodiment of the invention, in the formula I:

The R ₁ , the R ₂ , the R ₃ , the R ₄ , the R ₅ , the R ₆ , the R ₇ and the R ₈ are each independently hydrogen, —OH, —NH ₂ , halogen, optionally substituted C _1-5 linear or branched saturated or unsaturated hydrocarbon group, optionally substituted C _1-5 linear or branched alkoxy group, optionally substituted C _1-5 acyl group , optionally substituted phenyl,

or

Wherein R ₉ and R ₁₀ are each independently hydrogen, -OH, -NH ₂ , halogen, C _1-5 linear or branched saturated or unsaturated hydrocarbon group, carboxyl group, azide group, sulfhydryl group, amine group a base, a mercapto group, an amide group, a sulfate group, a rhamnose group, a phenyl group or a benzyl group, and optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH ₂ , Substituted by a group of phenyl, benzyl, heteroaryl, -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently saturated aliphatic C _1-4 alkyl, phenyl or benzyl The heteroaryl group is a 5- to 6-membered monocyclic heteroaryl group, and optionally contains 1 to 3 hetero atoms selected from N, O or S;

n is an integer of 2-7;

The halogen is -F, -Cl, -Br or -I.

According to an embodiment of the invention, in the formula I:

The R ₁ , the R ₂ , the R ₃ , the R ₄ , the R ₅ , the R ₆ , the R ₇ , the R ₈ and the R ₉ are each independently hydrogen, -OH, -NH ₂ , halogen, optionally substituted C _1-3 straight or branched saturated or unsaturated hydrocarbon group, optionally substituted C _1-3 straight or branched alkoxy group, optionally substituted C _1-3 acyl group, optionally substituted phenyl group,

or

Wherein R ₉ and R ₁₀ are each independently hydrogen, -OH, -NH ₂ , halogen, C _1-3 linear or branched saturated or unsaturated hydrocarbon group, carboxyl group, azide group, sulfhydryl group, amine group a base, a mercapto group, an amide group, a sulfate group, a rhamnose group, a phenyl group or a benzyl group, and optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH ₂ , Substituted by a group of phenyl, benzyl, heteroaryl, -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently saturated aliphatic C _1-4 alkyl, phenyl or benzyl a heteroaryl 5- to 6-membered monocyclic heteroaryl group, and optionally containing 1 hetero atom selected from N, O or S;

n is an integer of 2-7;

The halogen is -F, -Cl, -Br or -I.

According to an embodiment of the invention, in the formula I:

The R ₁ , the R ₂ , the R ₃ , the R ₄ , the R ₅ , the R ₆ , the R ₇ , and the R ₈ are each independently hydrogen, —OH, —NH ₂ , halogen, -OCH ₃ , -OCF ₃ , C _2-10 saturated aliphatic chain hydrocarbon, propenyl, propynyl, 2-(trifluoromethyl)ethyl, 2-bromoethyl, 2-azide Ethyl, acetyl, 2-halo(chloro, bromo)acetyl, 2-azidoacetyl, benzoyl, phenylacetyl, optionally substituted phenyl,

or

Wherein R ₉ and R ₁₀ are each independently hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, decyl, amine a base, a mercapto group, an amide group, a sulfate group, a rhamnose group, a phenyl group or a benzyl group, and the phenyl group and the benzyl group are optionally selected from one or more selected from the group consisting of halogen, -OCH ₃ , -OH, - Substituted by a group of OCF ₃ , —NH ₂ , —NHR ^X and —NR ^X ₂ , wherein each R ^{X is} independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

n is an integer of 2-7;

The halogen is -F, -Cl, -Br or -I.

According to an embodiment of the invention, the R ₁ is

The R ₂ , the R ₃ and the R ₄ are all hydrogen, the R ₅ is a methyl group, and the R ₇ and the R ₈ are both -OH, and the structural formula of the compound is as follows:

Wherein R ₆ is hydrogen, -OH, -NH ₂ , halogen, -OCH ₃ , -OCF ₃ , methyl, ethyl, isopropyl, vinyl, propenyl, propynyl, 2-(trifluoromethyl) Ethyl, 2-bromoethyl, 2-azidoethyl, acetyl, 2-halo(chloro, bromo)acetyl, 2-azidoacetyl, benzoyl, phenylacetyl or

Wherein R ₁₀ is hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, decyl, amine, sulfhydryl, amide a sulfate group, a rhamnosyl group, a phenyl group or a benzyl group, and the phenyl group and the benzyl group are optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH ₂ Substituting a group of -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

n is an integer of 2-7;

The halogen is -F, -Cl, -Br or -I.

According to an embodiment of the invention, the R ¹ is

R ₂ , R ₃ and R ₄ are all hydrogen, R ₅ is a methyl group, and R ₆ is -OH, and the structural formula of the compound is as follows:

Wherein R ₇ and R ₈ are each independently hydrogen, -OH, -NH ₂ , halogen, -OCH ₃ , -OCF ₃ , methyl, ethyl, isopropyl, vinyl, propenyl, propynyl, 2-(Trifluoromethyl)ethyl, 2-bromoethyl, 2-azidoethyl, acetyl, 2-halo(chloro, bromo)acetyl, 2-azidoacetyl, benzoyl, Phenylacetyl or

Wherein R ₉ is hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, sulfhydryl, amine, fluorenyl, amide a sulfate group, a rhamnosyl group, a phenyl group or a benzyl group, and the phenyl group and the benzyl group are optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH ₂ Substituting a group of -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

n is an integer of 2-7;

The halogen is -F, -Cl, -Br or -I.

According to an embodiment of the invention, the R ₂ is

R ₁ , R ₇ and R ₈ are both -OH, R ₃ and R ₅ are both hydrogen, and R ₄ is a methyl group. The structural formula of the compound is as follows, and the structural formula of the compound is as follows:

Wherein R ₆ is hydrogen, -OH, -NH ₂ , halogen, -OCH ₃ , -OCF ₃ , methyl, ethyl, isopropyl, vinyl, propenyl, propynyl, 2-(trifluoromethyl) Ethyl, 2-bromoethyl, 2-azidoethyl, acetyl, 2-halo(chloro, bromo)acetyl, 2-azidoacetyl, benzoyl, phenylacetyl or

Wherein R ₁₀ is hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, decyl, amine, sulfhydryl, amide a sulfate group, a rhamnosyl group, a phenyl group or a benzyl group, and the phenyl group and the benzyl group are optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH ₂ Substituting a group of -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

n is an integer of 2-7;

The halogen is -F, -Cl, -Br or -I.

According to an embodiment of the invention, the R ₂ is

R ₆ is -OH, R ₃ and R ₅ are all hydrogen, and R ₄ is a methyl group. The structural formula of the compound is as follows:

Wherein R ₁ , R ₇ , and R ₈ are each independently hydrogen, —OH, —NH ₂ , halogen, —OCH ₃ , —OCF ₃ , methyl, ethyl, isopropyl, vinyl, propenyl, and propyl. Alkynyl, 2-(trifluoromethyl)ethyl, 2-bromoethyl, 2-azidoethyl, acetyl, 2-halo(chloro,bromo)acetyl, 2-azidoacetyl, benzene Formyl, phenylacetyl or

Wherein R ₉ is hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, sulfhydryl, amine, fluorenyl, amide a sulfate group, a rhamnosyl group, a phenyl group or a benzyl group, and the phenyl group and the benzyl group are optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH ₂ , -NHR ^X and -NR ^X ₂ substituted groups, wherein, R ^X is independently saturated aliphatic C _1-4 alkyl, phenyl or benzyl;

n is an integer of 2-7;

The halogen is -F, -Cl, -Br or -I.

According to an embodiment of the invention, the R ₃ is

R ₁ , R ₇ and R ₈ are all -OH, R ₂ and R ₅ are all hydrogen, and R ₄ is a methyl group, and the structural formula of the compound is as follows:

Wherein R ⁶ is hydrogen, -OH, -NH ₂ , halogen, -OCH ₃ , -OCF ₃ , methyl, ethyl, isopropyl, vinyl, propenyl, propynyl, 2-(trifluoromethyl) Ethyl, 2-bromoethyl, 2-azidoethyl, acetyl, 2-halo(chloro, bromo)acetyl, 2-azidoacetyl, benzoyl, phenylacetyl or

Wherein R ₁₀ is hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, decyl, amine, sulfhydryl, amide a sulfate group, a rhamnosyl group, a phenyl group or a benzyl group, and the phenyl group and the benzyl group are optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH ₂ Substituting a group of -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

n is an integer of 2-7;

The halogen is -F, -Cl, -Br or -I.

According to an embodiment of the invention, the R ₃ is

R ₁ , R ₇ and R ₆ are both -OH, R ₂ and R ₅ are all hydrogen, and R ₄ is a methyl group. The structural formula of the compound is as follows:

Wherein, R1, R7 and R ₈ are each independently hydrogen, -OH, -NH _2, halo, -OCH _3, -OCF _3, methyl, ethyl, isopropyl, ethenyl, propenyl, propynyl , 2-(Trifluoromethyl)ethyl, 2-bromoethyl, 2-azidoethyl, acetyl, 2-halo(chloro,bromo)acetyl, 2-azidoacetyl, benzoyl Phenylacetyl or

Wherein R ₉ is hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, sulfhydryl, amine, fluorenyl, amide a sulfate group, a rhamnosyl group, a phenyl group or a benzyl group, and the phenyl group and the benzyl group are optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH ₂ Substituting a group of -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

n is an integer of 2-7;

The halogen is -F, -Cl, -Br or -I.

According to an embodiment of the invention, the polyether compound is one of the following compounds: R1 is

Or -OH, R2, R3 are each independently

Or hydrogen, R4 and R5 are each independently methyl or hydrogen, and R6, R7 and R8 are all -OH.

According to an embodiment of the invention, the polyether compound is one of the following compounds:

According to an embodiment of the present invention, the cancer is brain cancer, skin cancer, kidney cancer, bone cancer, sarcoma, prostate cancer, uterine cancer, melanoma, colon cancer, lymphoma, leukemia, pancreatic cancer, epithelial cancer, breast Cancer, liver cancer, lung cancer, stomach cancer, ovarian cancer and corresponding tumor cell stem cells.

According to an embodiment of the invention, the medicament prevents cancer recurrence by inhibiting tumor cell stem cell proliferation.

At present, the compound with polyether as the mother core is mainly used as an antibiotic, and the inventors have accidentally found that the compound has good antitumor activity through a large number of experiments. By mixing the three compounds with malignant tumor cells, they all have different degrees of antitumor activity. Thus, a compound having a polyether as a mother core can be used for treating malignant tumor cells in a pharmaceutically acceptable administration form (free form, sodium salt or potassium salt form, chelated form, hydrated form), enriching the treatment of such a compound The diversity of disease drugs. The inventors have found that the polyether compound represented by the general formula I can inhibit the proliferation of malignant tumor cells, and has studied three compounds of the compound 1, the compound 2 and the compound 3, and found that it has an inhibitory effect on various cancer cells. Specifically, the inventors have proved the inventor's point of view by the following experimental procedure:

(1) The inventors mixed and cultured various cancer cells with the three compounds of Compound 1, Compound 2 and Compound 3, especially the sodium salt thereof, in vitro. The inhibition rate and IC ₅₀ (half inhibition rate) of tumor cell proliferation by compound 1, compound 2 and compound 3 were calculated by MTT staining by measuring the OD value. The inventors have found that the three compounds of Compound 1, Compound 2 and Compound 3 have different degrees of inhibition on various tumor cells.

(2) The inventors found that the inhibitory effects of these three compounds on the proliferation of various cancer cells were significantly stronger than that of paclitaxel by comparison with the anticancer drug paclitaxel and salinomycin, which was found to have anticancer effects. The effect is partly stronger than salinomycin.

The additional aspects and advantages of the invention will be set forth in part in the description which follows.

DRAWINGS

The above and/or additional aspects and advantages of the present invention will become apparent and readily understood from

Figure 1 shows a schematic diagram of a compound 1 according to an embodiment of the present invention;

Figure 2 shows a schematic representation of a compound 2 in accordance with one embodiment of the present invention;

Figure 3 shows a schematic diagram of a compound 3 according to one embodiment of the present invention;

4 shows a schematic diagram of a flow pattern detection of stem cells in accordance with one embodiment of the present invention.

Detailed description of the invention

The embodiments described below with reference to the drawings are intended to be illustrative of the invention and are not to be construed as limiting.

Example 1

Compound 1, Compound 2 and Compound 3 were obtained by Streptomyces hygroscopicus Streptomyces and Streptomyces endus subtype Streptomyces, as follows:

Two Streptomyces hygroscopicus subtypes and Streptomyces endus subtype Streptomyces were inoculated on SFM plates and cultured for three to four days. When colonies are formed in the SFM medium, the single colony is cultured in the seed medium for about three days to four days, and the bacteria are inoculated to the fermentation medium in an amount of 1% (the composition of the fermentation medium is soluble starch 30 g/L, yellow). Bean cake powder 10g / L, yeast extract 2.5g / L, CaCO ₃ 3g / L, pH 7.2). Culture conditions: 30 degrees, 220 rotation speed, culture for 7 to 8 days.

When the culture is to be carried out on the 7th or 8th day, the fermentation broth is collected, and the supernatant is separated from the mycelium by high-speed centrifugation. The mycelium is added with the same amount of acetone, and the mixture is sonicated for 20 minutes, and the acetone and the mycelium are used. The amount of ethyl acetate was extracted and repeated once. The mixture was concentrated to dryness by rotary distillation, and then separated on a silica gel column to obtain crude product. The product was determined by HPLC. The HPLC results showed that the yield of the three strains was relatively high. Therefore, we also carried out TLC dot plate on the concentrated sample, and The color is developed in the iodine cylinder, which will be obviously smashed, dissolved in methanol, and centrifuged at high speed to take the mass spectrum to determine the position of the target point. The results of the compound detection are shown in Figure 1-3. Figure 1 shows that the compound is a compound. 1, Figure 2 shows that the compound is Compound 2, and Figure 3 shows that the compound is Compound 3.

Example 2

MTT assay was used to detect the inhibitory effect of compound 1 sodium salt and paclitaxel on MCF-7 tumor cells. The specific methods are as follows:

Take the logarithmic growth phase of MCF-7 (breast cancer cells, Wuhan University Basic Medical Research Institute Collection), and use 0.25%-EDTA trypsin digestion (3min-5min) when the cells are over 80% of the bottle wall. The cells are suspended. The cell suspension was inoculated into a 96-well cell culture plate (the number of cells per well was about 5000), 200 μL per well; the control group was a group of 200 μL of drug-free cells per well; the blank group was 200 μL per well. A group of cell-free media. The above-mentioned cells-inoculated 96-well cell culture plates were placed in a cell containing 5% CO ₂ incubator and incubated at 37 ° C overnight to allow the cells to adhere. Sample compound 1 sodium salt, salinomycin (salinomycin) and the existing anticancer drug paclitaxel were dissolved in DMSO, prepared as a 100 mg/mL stock solution, and then diluted to different concentrations with the corresponding medium before use (final concentration of DMSO was less than 0.1 %) sample solution. Add 200 μL of different concentrations of sample solution (Compound 1 sodium salt, salinomycin) (100 μg/mL, 10 μg/mL, 1 μg/mL, 0.1 μg/mL, 0.01 μg/mL) or paclitaxel solution to different experimental groups. (100 μg/mL, 10 μg/mL, 1 μg/mL, 0.1 μg/mL, 0.01 μg/mL); the control group was a group in which 200 μL of the medium was added to the adherent cells; the blank group was cell-free and only 200 μL of the medium was added. group. Then, the 96-well cell culture plate supplemented with different concentrations of the drug solution (100, 10, 1, 0.1, 0.01 μg / L sample solution or salinomycin or paclitaxel solution) was cultured in a cell culture incubator (37 ° C) for 48 hours. Thereafter, 20 μL of 0.5 mg/mL of thiazolyl blue (MTT) was added to each well of the above-mentioned 96-well cell culture plate, and then cultured in a cell culture incubator (37 ° C). After 2-4 hours, 96-well cell culture plates co-incubated with MTT were centrifuged at 2000 rpm for 10 minutes in a high speed centrifuge. After centrifugation, the supernatant in a 96-well cell culture plate was removed, and then the formazan blue crystals formed at the bottom of the 96-well plate were dissolved by adding 100 μL of dimethyl sulfoxide (DMSO) to each well. The 96-well cell culture plate with DMSO was shaken on a plate shaker for 10 minutes, and the OD value (reference wavelength was 490 nm) was measured at 570 nm using an enzyme-linked immunosorbent assay to calculate the inhibition rate and IC ₅₀ (half number). Inhibition rate).

Inhibition rate%=(AA ₀ )/(AA ₁ )×100%

Where: A represents the OD value of the control group; A ₀ represents the OD value of the sample group; A ₁ represents the OD value of the blank group.

IC ₅₀ values of compound 1 sodium salt and paclitaxel on MCF-7 cells

Test substance	IC 50 (μmol/L) for MCF-7
Compound 1 sodium salt	1.63±0.04
Paclitaxel	8.49±1.43
Salinomyicn	3.86±0.08

From the results shown in the above table, compared with paclitaxel, the sodium salt of Compound 1 has a more significant inhibitory effect on the proliferation of MCF-7 cells in vitro, and the IC ₅₀ value of MCF-7 cells is 5.21 times lower than that of paclitaxel. Compared with salinomycin, it was 2.34 times lower. It indicated that the sodium salt of compound 1 had a strong inhibitory effect on the proliferation of MCF-7 tumor cells, and its inhibitory effect was significantly stronger than that of paclitaxel.

Example 3:

The inhibitory effect of Compound 2 and paclitaxel on MCF-7 tumor cells was examined by MTT assay. The experimental method and detection method were the same as those in Example 2, except that the compound used was Compound 2. The sodium salt of Compound 2 and the IC ₅₀ value of paclitaxel to MCF-7 cells are shown in the following table.

And taxol sodium salt of compound 2 IC ₅₀ values for the MCF-7 cells

Test substance	IC 50 (μmol/L) for MCF-7
Compound 2 sodium salt	2.25±0.19
Paclitaxel	8.49±1.43
Salinomyicn	3.86±0.08

From the results shown in the above table, compared with salinomycin and paclitaxel, the sodium salt of compound 2 has a more significant inhibitory effect on the proliferation of MCF-7 cells in vitro, and the IC ₅₀ value of MCF-7 cells is lower than that of paclitaxel. It was reduced by 3.77 times, which is equivalent to salinomycin. It indicated that the sodium salt of compound 2 had a strong inhibitory effect on the proliferation of MCF-7 tumor cells, and its inhibitory effect was significantly stronger than that of paclitaxel.

Example 4:

The inhibitory effect of Compound 3 and paclitaxel on MCF-7 tumor cells was examined by MTT assay. The experimental method and detection method were the same as those in Example 2, except that the compound used was Compound 3. The sodium salt of Compound 3 and the IC ₅₀ value of paclitaxel to MCF-7 cells are shown in the following table.

Test substance	IC 50 (μmol/L) for MCF-7
Compound 3 sodium salt	5.19±1.4
Paclitaxel	8.49±1.43
Salinomyicn	3.86±0.08

From the results shown in the table, the compound 3 sodium salt has a more significant inhibitory effect on the proliferation of MCF-7 cells in vitro than the paclitaxel, and the IC ₅₀ value of the MCF-7 cell line is 1.61 times lower than that of paclitaxel. It indicated that the sodium salt of compound 4 had a strong inhibitory effect on the proliferation of MCF-7 tumor cells, and its inhibitory effect was stronger than that of paclitaxel, but its anticancer activity was inferior to that of salinomycin.

Example 5:

The preparation method of the compound sodium salt derivative compound of the general formula of the present invention is specifically as follows:

Compounds 1, 2, 3 (1.7 g, 200 mmol), dicyclohexylcarbodiimide (240 mmol) and 4-dimethylaminopyridine (10 m, respectively) at 0 ° C under anhydrous anaerobic conditions The mixture was dissolved in dichloromethane (8 ml), and the corresponding reagent was slowly added dropwise with stirring, and the reaction was allowed to stand at room temperature overnight after the completion of the dropwise addition. The reaction system was quenched by the addition of a 0.1 mol/L HCl solution, and the reaction mixture was washed with brine, and extracted with ethyl acetate. The organic phase was combined and dried over anhydrous sodium sulfate. The concentrate is passed through a column to obtain a structural derivative of the formula II, IV, VI.

Example 6

The preparation method of the compound sodium salt derivative compound of the general formula of the present invention is specifically as follows:

Compounds 1, 2, 3 (1.7 g, 200 mmol), and 4-dimethylaminopyridine (10 mmol) were dissolved in dichloromethane (8 ml) at 0 ° C under anhydrous anaerobic conditions. Triethylamine (600 mmol) was slowly added dropwise with stirring, and after stirring for 10 min, the corresponding acid chloride and brominated compound were further added dropwise. After the completion of the dropwise addition, the reaction was allowed to stand at room temperature for several hours. The reaction system was quenched by the addition of a 0.1 mol/L HCl solution, and the reaction mixture was washed with brine, and extracted with ethyl acetate. The organic phase was combined and dried over anhydrous sodium sulfate. The concentrate is passed through a column to obtain a derivative of the structure shown in Formula III, V, and VII.

Example 7

The inhibitory effects of compounds 1, 2, and 3 on various tumor cells were examined as follows:

(1) Cell resuscitation: Before the experiment, the ultra-clean table top was irradiated with ultraviolet rays for 30 minutes. The water bath was preheated to 37 ° C and the freshly prepared medium was preheated in a water bath. The frozen cells are taken out, and the frozen tube is quickly thawed into a preheated water bath and continuously shaken to rapidly melt the liquid in the tube. After about 1-2 minutes, the liquid in the cryotube was completely dissolved, and the outer wall of the frozen tube was wiped out with a 70% alcohol cotton ball. Pipette the cells in the cryotube and transfer to a 15 ml centrifuge tube while adding 5 ml of pre-warmed complete medium. Centrifuge at 500g for 3-5min at low speed and aspirate the supernatant. 10 ml of the culture solution was added to the centrifuge tube, and gently pipetted to prepare a cell suspension. After counting the cells by trypan blue staining and measuring the viability, the cell suspension was added to a 10 cm culture dish and cultured overnight in a 37 ° C / 5% CO 2 incubator.

(2) Cell culture: The ratio of the medium and passage required for cell culture is referred to ATCC. During the subculture process before the cell experiment, the medium of the cell is replaced with phenol-free red medium + 10% FBS.

(3) Cell inhibition experiment: Compound samples were diluted with DMSO at a concentration of 5 times in a concentration of 5 times, and diluted with a complete medium without phenol red to prepare a solution for experimental concentration (5 times final concentration).

Cell viability assay method:

a) Inoculation of cells: Log phase cells were collected, the cell suspension concentration was adjusted, 40 μl of the cell suspension was inoculated in a 384-well plate, and the marginal wells were filled with sterile PBS.

b) The cell plates were incubated overnight in a 37 ° C / 5% CO 2 incubator, and 10 μL of a compound sample of 5 times the concentration to be tested was added the next day.

c) Cells were incubated in a 37 ° C / 5% CO ₂ incubator and observed with an inverted microscope for 72 hours.

d) Plate reading: After 72 hours, incubate for 10 min at room temperature, add 30 μL per well of CellTiter-Glo reaction mixture per well, shake the assay plate for 2-3 minutes, and incubate for 10 min at room temperature. Read the RLU value at Pherastar (BMG labtech) and save the data.

Cell Growth inhibition%=100%×[1-RLU sample/RLU negative], wherein the RLU sample is the sample well or the positive control well RLU value, and the RLU negative is the DMSO-only RLU value. Data analysis was performed using GraphPad Prism 6.0 software, and the results are shown in the following table, indicating that the compound has a significant inhibitory effect on various tumors.

IC ₅₀ values of Compound 1, Compound 2 and Compound 3 against various tumor cells

Example 8

The inhibitory effects of Compound 1, Compound 2 and Compound 3 on the proliferation of tumor stem cells were examined. The specific methods are as follows:

(1) Determination of the content of enriched stem cells in suspension culture:

Cell culture subject: human breast cancer cell line MDA-MB-231 cells.

Culture method: cultured in suspension cell culture, cultured in a 37-degree incubator.

Suspension cell culture medium (200 ml): B27 4 ml; insulin 80 μl; DMEM-F12 194 ml; hEGF: 40 μl; double antibody: 2 ml

Culture flask: T25 (15ml); T75 (75ml);

Inoculation density: 1000 / ml;

Test kit: Aldefluor ^TM kit;

Cultivation time: 10 days to 15 days or so (shake more than 2 times a day).

experiment procedure:

(1) After the cells are cultured, they are dispensed into a 15 ml centrifuge tube, and the T25 bottle is washed once, and the mixture is centrifuged at 250 g for 10 minutes;

(2) Remove the supernatant, add 1ml (0.05%) of trypsin, wash once with PBS, transfer to T25 flask, slowly gently hit the single cell process for 5min to 10min;

(3) transferred to a 15ml centrifuge tube, and centrifuged 250g for 10min;

(4) Add 1 ml of PBS in two portions, transfer to a 1.5 ml EP tube, and evenly beat and count a small amount;

(5) Calculate the number in the 100,000 to 200,000 cells and the EP tube, remove the PBS by centrifugation, and resuspend in 1 ml of the kit buffer;

(6) Add 10 μl of DEAB to another EP tube, then add 10 μl of activated Aldfluor reagent to the EP tube of the cell, mix well quickly, pipet 0.5 ml into the EP tube of DEAB, mix well, and place in a 37-degree incubator. Medium culture for 45 min;

(7) After 45 min, the supernatant was removed by centrifugation (250 g, 10 min), resuspended in 0.5 ml of reagent buffer, placed on ice and stored in the dark.

The cells were detected by flow cytometry. The results of the experiment are shown in Fig. 4. The content of stem cells in adherent cells was 2.36%, and the proportion of stem cells obtained after suspension cell culture was 25.95%, which was nearly 11 times higher. There are significant differences between the data. It can be seen that we have achieved the goal of enriching stem cells.

(2) Drug inhibition test:

Drugs: Compound 1, Compound 2, and Compound 3

Cell type: step (1) enriched stem cells

Cell seeding: The amount of cells per well per well of 96-well cells was about 12,000 per 120 μL.

Dilution concentration of drug: diluted with PBS: setting concentration: 100 μg/ML, 20 μg/ML, 4 μg/ML, 0.8 μg/ML, 0.16 μg/ML, 0.032 μg/ML, 0.0064 μg/ML, 0.00128 μg/ML, 0.000256 Gg/ML.

Addition of the compound: The cell plate was incubated overnight at 37 ° C / 5% CO 2 incubator, and 10 μL of a compound sample of 5 times the concentration to be tested was added the next day.

Detection time and detection method: After 72 hours of administration, the MTT method was used for detection, and then the triple solution (10% SDS, 5% isobutanol, 0.1% concentrated hydrochloric acid) was used for dissolution, and the OD value was detected at 570 nm by a microplate reader.

Each compound was repeated independently. The results are as follows

Test drug	IC 50 (g/ml)
Compound 1	1.176E-06±0.041
Compound 2	2.35E-06±0.152
Compound 3	4.251E-06±0.129

The results showed that Compound 1, Compound 2 and Compound 3 all had significant inhibitory effects on tumor stem cells, which could inhibit tumor recurrence.

In the description of the present specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" and the like means a specific feature described in connection with the embodiment or example. A structure, material or feature is included in at least one embodiment or example of the invention. In the present specification, the schematic representation of the above terms is not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in a suitable manner in any one or more embodiments or examples. In addition, various embodiments or examples described in the specification, as well as features of various embodiments or examples, may be combined and combined.

Although the embodiments of the present invention have been shown and described, it is understood that the above-described embodiments are illustrative and are not to be construed as limiting the scope of the invention. The embodiments are subject to variations, modifications, substitutions and variations.

Claims (14)

1. Use of a polyether compound of the formula I, a chelated form thereof, a hydrated form or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of cancer and for the prevention of cancer recurrence,

   

   among them,

   The R ₁ , the R ₂ , the R ₃ , the R ₄ , the R ₅ , the R ₆ , the R ₇ and the R ₈ are each independently hydrogen, —OH, —NH ₂ , halogen, optionally substituted C _1-10 linear or branched saturated or unsaturated hydrocarbon group, optionally substituted C _1-10 linear or branched alkoxy group, optionally substituted C _1-10 acyl group , optionally substituted C _{6 ~ 20} aryl group, an optionally substituted heteroaryl group,

   

   Wherein R ₉ and R ₁₀ are each independently hydrogen, -OH, -NH ₂ , halogen, C _1-5 linear or branched saturated or unsaturated hydrocarbon group, carboxyl group, azide group, fluorenyl group , amine, mercapto, amido, sulfate, rhamnose, phenyl or benzyl, and optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH _2, phenyl, benzyl, heteroaryl, -NHR ^X and -NR ^X ₂ substituted groups, wherein, R ^X is independently saturated aliphatic C _1-4 alkyl, phenyl or a benzyl group, the heteroaryl group being a 3- to 6-membered monocyclic or bicyclic heteroaryl group, and optionally containing 1 to 3 hetero atoms selected from N, O or S;

   n is an integer of 2-7;

   The halogen is -F, -Cl, -Br or -I.
2. The use according to claim 1, characterized in that, in the formula I:

   The R ₁ , the R ₂ , the R ₃ , the R ₄ , the R ₅ , the R ₆ , the R ₇ and the R ₈ are each independently hydrogen, —OH, —NH ₂ , halogen, optionally substituted C _1-5 linear or branched saturated or unsaturated hydrocarbon group, optionally substituted C _1-5 linear or branched alkoxy group, optionally substituted C _1-5 acyl group , optionally substituted phenyl,

   

   

   Wherein R ₉ and R ₁₀ are each independently hydrogen, -OH, -NH ₂ , halogen, C _1-5 linear or branched saturated or unsaturated hydrocarbon group, carboxyl group, azide group, fluorenyl group , amine, mercapto, amido, sulfate, rhamnose, phenyl or benzyl, and optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH _2, phenyl, benzyl, heteroaryl, -NHR ^X and -NR ^X ₂ substituted groups, wherein, R ^X is independently saturated aliphatic C _1-4 alkyl, phenyl or a benzyl group, the heteroaryl group being a 5- to 6-membered monocyclic heteroaryl group, and optionally containing 1 to 3 hetero atoms selected from N, O or S;

   n is an integer of 2-7;

   The halogen is -F, -Cl, -Br or -I.
3. The use according to claim 2, characterized in that in the formula I:

   The R ₁ , the R ₂ , the R ₃ , the R ₄ , the R ₅ , the R ₆ , the R ₇ and the R ₈ are each independently hydrogen, —OH, —NH ₂ , halogen, optionally substituted C _1-3 linear or branched saturated or unsaturated hydrocarbon group, optionally substituted C _1-3 linear or branched alkoxy group, optionally substituted C _1-3 acyl group , optionally substituted phenyl,

   

   

   Wherein R ₉ and R ₁₀ are each independently hydrogen, -OH, -NH ₂ , halogen, C _1-3 linear or branched saturated or unsaturated hydrocarbon group, carboxyl group, azide group, fluorenyl group , an amine group, a thiol group, an amide group, a sulfate group, a rhamnose group, a phenyl group or a benzyl group, and optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , - Substituted by a group of NH ₂ , phenyl, benzyl, heteroaryl, -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently a saturated aliphatic C _1-4 alkyl group, phenyl group Or a benzyl group, the heteroaryl group being a 5- to 6-membered monocyclic heteroaryl group, and optionally containing 1 hetero atom selected from N, O or S;

   n is an integer of 2-7;

   The halogen is -F, -Cl, -Br or -I.
4. The use according to claim 3, characterized in that in the formula I:

   The R ₁ , the R ₂ , the R ₃ , the R ₄ , the R ₅ , the R ₆ , the R ₇ , and the R ₈ are each independently hydrogen, —OH, —NH ₂ , halogen, -OCH ₃ , -OCF ₃ , C _2-10 saturated aliphatic chain hydrocarbon, propenyl, propynyl, 2-(trifluoromethyl)ethyl, 2-bromoethyl, 2-azide Ethyl, acetyl, 2-halo(chloro, bromo)acetyl, 2-azidoacetyl, benzoyl, phenylacetyl, optionally substituted phenyl,

   

   

   Wherein R ₉ and R ₁₀ are each independently hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, fluorenyl An amine group, a mercapto group, an amide group, a sulfate group, a rhamnose group, a phenyl group or a benzyl group, and the phenyl group and the benzyl group are optionally selected from one or more selected from the group consisting of halogen, -OCH ₃ , -OH Substituted by a group of -OCF ₃ , -NH ₂ , -NHR ^X and -NR ^X ₂ , wherein each R ^{X is} independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

   n is an integer of 2-7;

   The halogen is -F, -Cl, -Br or -I.
5. The use according to claim 4, wherein said R ₁ is

   

   The R ₂ , the R ₃ and the R ₄ are all hydrogen, the R ₅ is a methyl group, and the R ₇ and the R ₈ are both -OH, and the structural formula of the compound is as follows:

   

   Wherein R ₆ is hydrogen, -OH, -NH ₂ , halogen, -OCH ₃ , -OCF ₃ , methyl, ethyl, isopropyl, vinyl, propenyl, propynyl, 2-(trifluoromethyl) Ethyl, 2-bromoethyl, 2-azidoethyl, acetyl, 2-halo(chloro, bromo)acetyl, 2-azidoacetyl, benzoyl, phenylacetyl or

   

   Wherein R ₁₀ is hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, decyl, amine, sulfhydryl, amide a sulfate group, a rhamnosyl group, a phenyl group or a benzyl group, and the phenyl group and the benzyl group are optionally selected from one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , Substituted by a group in NH ₂ , —NHR ^X and —NR ^X ₂ , wherein R ^{X is} each independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

   n is an integer of 2-7;

   The halogen is -F, -Cl, -Br or -I.
6. The use according to claim 4, wherein said R ₁ is

   

   R ₂ , R ₃ and R ₄ are all hydrogen, R ₅ is a methyl group, and R ₆ is -OH, and the structural formula of the compound is as follows:

   

   Wherein R ₇ and R ₈ are each independently hydrogen, -OH, -NH ₂ , halogen, -OCH ₃ , -OCF ₃ , methyl, ethyl, isopropyl, vinyl, propenyl, Propynyl, 2-(trifluoromethyl)ethyl, 2-bromoethyl, 2-azidoethyl, acetyl, 2-halo(chloro,bromo)acetyl, 2-azidoacetyl, Benzoyl, phenylacetyl or

   

   Wherein R ₉ is hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, sulfhydryl, amine, fluorenyl, amide a sulfate group, a rhamnosyl group, a phenyl group or a benzyl group, and the phenyl group and the benzyl group are optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH ₂ Substituting a group of -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

   n is an integer of 2-7;

   The halogen is -F, -Cl, -Br or -I.
7. The use according to claim 4, wherein said R ₂ is

   

   The R ₁ , the R ₇ and the R ₈ are both -OH, the R ₃ and the R ₅ are both hydrogen, the R ₄ is a methyl group, and the compound has the following structural formula, the compound The structure is as follows:

   

   Wherein R ₆ is hydrogen, -OH, -NH ₂ , halogen, -OCH ₃ , -OCF ₃ , methyl, ethyl, isopropyl, vinyl, propenyl, propynyl, 2-(three Fluoromethyl)ethyl, 2-bromoethyl, 2-azidoethyl, acetyl, 2-halo(chloro,bromo)acetyl, 2-azidoacetyl, benzoyl, phenylacetyl or

   

   Wherein R ₁₀ is hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, decyl, amine, sulfhydryl, amide , sulfate groups, rhamnose group, phenyl or benzyl, and the phenyl and benzyl optionally substituted with one or more substituents selected from _{halo, -OCH 3, -OH, -OCF 3} , -NH 2 Substituting a group of -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

   n is an integer of 2-7;

   The halogen is -F, -Cl, -Br or -I.
8. The use according to claim 4, wherein said R ₂ is

   

   The R ₆ is -OH, R ₃ and R ₅ are both hydrogen, and R ₄ is a methyl group. The structural formula of the compound is as follows:

   

   Wherein R ₁ , R ₇ and R ₈ are each independently hydrogen, -OH, -NH ₂ , halogen, -OCH ₃ , -OCF ₃ , methyl, ethyl, isopropyl, ethylene Base, propenyl, propynyl, 2-(trifluoromethyl)ethyl, 2-bromoethyl, 2-azidoethyl, acetyl, 2-halo(chloro, bromo)acetyl, 2-fold Nitroacetyl, benzoyl, phenylacetyl or

   

   Wherein R ₉ is hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, sulfhydryl, amine, fluorenyl, amide , sulfate groups, rhamnose group, phenyl or benzyl, and the phenyl and benzyl optionally substituted with one or more substituents selected from _{halo, -OCH 3, -OH, -OCF 3} , -NH 2 Substituting a group of -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

   n is an integer of 2-7;

   The halogen is -F, -Cl, -Br or -I.
9. The use according to claim 4, wherein said R ₃ is

   

   The R ₁ , the R ₇ and the R ₈ are both -OH, the R ₂ and the R ₅ are hydrogen, and the R ₄ is a methyl group, and the structural formula of the compound is as follows:

   

   Wherein R ₆ is hydrogen, -OH, -NH ₂ , halogen, -OCH ₃ , -OCF ₃ , methyl, ethyl, isopropyl, vinyl, propenyl, propynyl, 2-(three Fluoromethyl)ethyl, 2-bromoethyl, 2-azidoethyl, acetyl, 2-halo(chloro,bromo)acetyl, 2-azidoacetyl, benzoyl, phenylacetyl or

   

   Wherein R ₁₀ is hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, decyl, amine, sulfhydryl, amide a sulfate group, a rhamnosyl group, a phenyl group or a benzyl group, and the phenyl group and the benzyl group are optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH ₂ Substituting a group of -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

   n is an integer of 2-7;

   The halogen is -F, -Cl, -Br or -I.
10. The use according to claim 4, wherein said R ₃ is

    

    The R ₁ , the R ₇ and the R ₆ are both -OH, the R ₂ and the R ₅ are both hydrogen, and the R ₄ is a methyl group, and the structural formula of the compound is as follows:

    

    Wherein R ₁ , R ₇ and R ₈ are each independently hydrogen, -OH, -NH ₂ , halogen, -OCH ₃ , -OCF ₃ , methyl, ethyl, isopropyl, ethylene Base, propenyl, propynyl, 2-(trifluoromethyl)ethyl, 2-bromoethyl, 2-azidoethyl, acetyl, 2-halo(chloro, bromo)acetyl, 2-fold Nitroacetyl, benzoyl, phenylacetyl or

    

    Wherein R ₉ is hydrogen, -OH, -NH ₂ , halogen, methyl, ethyl, isopropyl, vinyl, propenyl, carboxyl, azide, sulfhydryl, amine, fluorenyl, amide a sulfate group, a rhamnosyl group, a phenyl group or a benzyl group, and the phenyl group and the benzyl group are optionally one or more selected from the group consisting of halogen, -OCH ₃ , -OH, -OCF ₃ , -NH ₂ Substituting a group of -NHR ^X and -NR ^X ₂ wherein R ^{X is} each independently a saturated aliphatic C _1-4 alkyl group, a phenyl group or a benzyl group;

    n is an integer of 2-7;

    The halogen is -F, -Cl, -Br or -I.
11. The use according to claim 4, wherein the polyether compound is one of the following compounds: the R ₁ is

    

    Or -OH, the R ₂ and the R ₃ are each independently

    

    Or hydrogen, the R ₄ and the R ₅ are each independently methyl or hydrogen, and the R ₆ , the R ₇ and the R ₈ are both -OH.
12. The use according to any one of claims 1 to 11, wherein the polyether compound is one of the following compounds:

    
13. The use according to claim 1, wherein the cancer is brain cancer, skin cancer, kidney cancer, bone cancer, sarcoma, prostate cancer, uterine cancer, melanoma, colon cancer, lymphoma, leukemia, pancreatic cancer. , epithelial cell carcinoma, breast cancer, liver cancer, lung cancer, stomach cancer and ovarian cancer.
14. The use according to claim 1, wherein the drug prevents cancer recurrence by inhibiting proliferation of tumor cell stem cells.

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