WO2008031358A1

WO2008031358A1 - 1,2-dihydroxy-3-methyl-9,10-anthraquinone in the manufacture of medicaments for treating cancers

Info

Publication number: WO2008031358A1
Application number: PCT/CN2007/070650
Authority: WO
Inventors: Long Yu; Haoxing Zhang; Qingyu Lang; Lisha Tang; Jia Li
Original assignee: Fudan University
Priority date: 2006-09-07
Filing date: 2007-09-07
Publication date: 2008-03-20
Also published as: CN101112368A

Abstract

The use of 1,2-dihydroxy-3-methyl-9,10-anthraquinone in the manufacture of medicaments for treating cancers. The compound demonstrates inhibition towards liver, colon, lung and stomach cancer cells and phosphokinase Aurora-B. The compound of the present invention can be formulated into anticancer pharmaceutical compositions with pharmaceutically acceptable carriers to suppress the continuous growth of tumors.

Description

Application of 1,2-sepi-3-methyl-9,10-indole in preparation of anticancer drugs

The present invention relates to the field of cell biology, and provides the use of the compound s4 for the preparation of an anticancer drug. Background technique

Tumors are diseases that seriously endanger human health. The incidence of cancer has increased in recent years. At present, more than 6 million people worldwide die of cancer every year, and the number of new cases is 8 million. This number is increasing year by year. There are millions of new cancer patients in China each year, with about 2 million patients to be treated and about 1.3 million deaths. It is estimated that by the end of 2006, 1.7 million people will die of cancer in China, and cancer has become the first cause of death.

Malignant tumors, commonly known as cancer, are unrestricted and diffuse masses caused by the mutating of organs and tissue cells of the human body. It can destroy the organs and tissues of the human body, consumes limited energy in the body, and damages human life and even causes death.

Mitosis is a very important stage in the cell cycle process. The difference between a tumor and a normal cell is unrestricted proliferation, that is, mitosis loses its normal regulation. Most tumors occur because of the instability of the genomic group caused by mitosis. The instability of the genome is also derived from the instability of the chromosome. The instability of the chromosome is a characteristic marker of cancer. Abnormal chromosome segregation, central granule replication and segregation can lead to abnormalities in the number of chromosomes per cell, ie aneuploidy, increased chance of heterozygous deletion, loss of tumor suppressor gene or increased number of proto-oncogenes Disrupts cell life metabolism and signaling, increasing the chance of cancer. Kinase plays an important role in the completion of mitosis and cell cycle regulation, and thus has become a research hotspot in molecular biology.

In order to treat tumors, there is an urgent need in the art to develop compounds capable of effectively inhibiting the growth of tumor cells. Summary of the invention

It is an object of the present invention to provide the use of the compound s4 for the preparation of an anticancer drug. In a first aspect of the invention, there is provided a compound of formula I, ie 1,2-dihydroxy-3-methyl-9, 10-oxime, or a pharmaceutically acceptable salt or ester thereof Use in the preparation of anti-cancer compositions such as anti-cancer drugs.

In another preferred embodiment, the composition is a pharmaceutical composition, a nutraceutical composition or a dietary supplement composition.

In another preferred embodiment, the medicament is for killing cancer cells or inhibiting the growth of cancer cells.

In another preferred embodiment, the anticancer drug contains 0.001 to 99% by weight (; preferably 0.01 to 90% by weight;) a compound of the formula I and a pharmaceutically acceptable carrier or excipient.

In another preferred embodiment, the medicament is also for inhibiting phosphokinase Aurora-B.

In another preferred embodiment, the anticancer drug is an injection, granule, capsule, solution or tablet.

In another preferred embodiment, the cancer is selected from the group consisting of liver cancer, colon cancer, gastric cancer, and lung cancer.

In a second aspect of the present invention, a method for inhibiting growth of a tumor cell in vitro is provided, comprising the steps of: adding a compound 1,2-dihydroxy-3-methyl-9, 10- in a culture system of the tumor cell Hey.

In another preferred embodiment, the tumor cells are selected from the group consisting of liver cancer cells, colon cancer cells, gastric cancer cells, and lung cancer cells.

In a third aspect of the invention, a method of treating cancer comprising the steps of: administering 1,2-dihydroxy-3-methyl-9, 10-oxime or a pharmaceutically thereof to a mammalian subject in need of treatment Acceptable salt.

In another preferred embodiment, the application amount is from 0.1 mg/kg to 20 mg/kg of body weight per day. DRAWINGS

Figure 1 is a graph showing the inhibition results of s4 against _SW 620 cells. It can be seen that the IC50 (half lethal) of the s4 of the present invention against the human colon cancer cell line (sw620) is about 1 μΜ. detailed description

The present invention provides the use of the compound represented by the following formula (I) in the preparation of an anticancer drug

The compound has the name 1,2-dihydroxy-3-methylanthra-9, 10- qui e [ 1 2-dihydroxy-3-methyl-9 10-oxime], abbreviated as s4, and has a molecular weight of 254. It is prepared by a commercially available or conventional method. The invention also includes compounds obtained by conventional substitution of the compounds.

The s4 compound of the present invention can be used in the form of a salt or ester derived from a pharmaceutically or physiologically acceptable acid or base. These salts include, but are not limited to, salts or esters formed with: hydrochloric acid, hydrobromic acid, sulfuric acid, citric acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, succinic acid, oxalic acid, fumaric acid, Maleic acid, oxaloacetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, or isethionic acid. The salts of halides are also suitable. Other salts include: salts with alkali or alkaline earth metals such as sodium, potassium, calcium or magnesium, as well as esters, carbamates or other conventional "prodrugs" (when given in this form) In the case of medicine, it can be converted into an active part in the body). In the present invention, the compound s4 can be used for the preparation of an anticancer drug.

The present invention detects the action of the compound s4 on tumor cells and cancer cells according to a standard MTS assay, and the results show that the s4 of the present invention is applicable to various tumor cells and cancer cells, such as liver cancer cells, colon cancer cells, gastric cancer cells, lung cancer cells, and the like. , have a killing or inhibition effect.

Further studies have shown that the compound s4 of the present invention has an inhibitory effect on the phosphokinase Aurora-B. This further confirmed that the compound s4 of the present invention has an effect of inhibiting malignant tumors.

Malignant tumors, commonly known as cancer, are unrestricted and diffuse masses caused by the mutating of organs and tissue cells of the human body. Kinase plays an important role in the completion of mitosis and cell cycle regulation, and thus has become a research hotspot in molecular biology. AIK (Aur ₀ ra/Ipl l Kinase ) is one of the five major regulatory kinase families currently known. So far, the human Aiks family has found three members: Aikl Aik2 (Aurora-B) and Aik3. Among them, Aik2 mainly plays a role in the mitosis of cells. It acts as a chromosomal concomitant protein involved in the regulation of the interaction between the centromere and the spindle microtubule. At the metaphase to the late stage, Aik2 migrates from the centromere to the middle zone of the cell, and is distributed at the end of the cell plate and the posterior mitotic bridge. Post-mitotic event, so staining The separation of the body is related to cytokinesis. Mutations in Aik2 inhibit the formation of dividing sulcus and lead to the formation of multinucleated cells. Aik2 is localized to 17ql3, an area that is abnormal in a variety of human cancer cells, and Aik2 is highly expressed in abnormally ploidy colon cancer cell lines. Therefore, researchers are looking for molecules that interact with Aik2 and use them as antitumor drugs.

The present invention also establishes a mouse model of transplanted tumor for detecting the effect of s4 on tumors and cancer cells in vivo. The results also show that s4 can inhibit tumor growth in vivo. In the present invention, the anticancer drug prepared from the compound s4 is a pharmaceutical composition comprising a therapeutically effective amount of the compound s4 and a pharmaceutically acceptable carrier or excipient.

In the present invention, the effective application amount is 0.1 mg / kg to 20 mg / kg body weight per day. In the present invention, the antitumor drug prepared may be an injection or a tablet or the like.

The present invention, s4, its analogs and the like, can provide different effects when administered (administered) therapeutically. Generally, these materials can be formulated in a non-toxic, inert, and pharmaceutically acceptable aqueous carrier medium wherein the pH is usually from about 5 to about 8, preferably from about 6 to about 8, although the pH may be The nature of the formulation and the condition to be treated vary. The formulated pharmaceutical compositions can be administered by conventional routes including, but not limited to, intramuscular, intraperitoneal, subcutaneous, intradermal, intratumoral, or topical administration.

Taking s4 of the present invention as an example, it can be used in combination with a suitable pharmaceutically acceptable carrier. Such pharmaceutical compositions contain a therapeutically effective amount of the compound and a pharmaceutically acceptable carrier or excipient. Such carriers include, but are not limited to, saline, buffer, dextrose, water, glycerol, ethanol, and combinations thereof. The pharmaceutical preparation should be matched to the mode of administration. The s4 of the present invention can be prepared in the form of an injection, for example, by a conventional method using physiological saline or an aqueous solution containing glucose and other adjuvants. Pharmaceutical compositions such as tablets and capsules can be prepared by conventional methods. Pharmaceutical compositions such as injections, solutions, tablets and capsules are preferably prepared under sterile conditions. The active ingredient is administered in a therapeutically effective amount, for example, from about 1 microgram per kilogram body weight to about 5 milligrams per kilogram body weight per day. Furthermore, s4 of the invention may also be used with other therapeutic agents.

When s4 of the invention is used as a medicament, a therapeutically effective amount of s4 can be administered to a mammal, wherein the therapeutically effective dose is typically at least about 10 micrograms per kilogram of body weight, and in most cases no more than about 80 milligrams per kilogram. The body weight, preferably the dose is from about 500 micrograms per kilogram of body weight to about 2 milligrams per kilogram of body weight. Of course, specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician. Cancer has become the first cause of death, and the substances that kill tumors and cancer cells have become the research hotspots in the biomedical field. The s4 of the present invention has a killing or inhibiting effect on various tumor cells and cancer cells; meanwhile, the compound s4 of the present invention has an inhibitory effect on the phosphokinase Aurora-B. This all proves that the compound s4 of the present invention has an effect of inhibiting malignant tumors. The compound s4 of the present invention can be used in combination with a suitable pharmaceutically acceptable carrier to prepare an anticancer drug to inhibit the continued growth of the tumor. The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are merely illustrative of the invention and are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually carried out according to the conditions described in conventional conditions such as Sambrook et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer. The suggested conditions. Example 1 In vitro enzyme activity inhibition experiment

The human Aurora-B protein kinase (NCBI accession number NP-004208) was cloned, expressed and purified using E. coli expression system for high-throughput screening experiments. Using radioisotope technology, AURORAB phosphorylation substrate MBP (UPSTATE, CAT#13-110, lot#27845), covalently binds γ- ³³ [ on the [γ- ³³ Ρ]-ΑΤΡ in solution to the substrate. Its Ser is phosphorylated to produce the substrate ³³ P-MBP with isotopic labeling. Finally, the activity of the enzyme was detected by counting the scintillation liquid by counting with a liquid scintillation counter, and the inhibition of the activity of the different compounds was observed to initially evaluate the activity of the compound.

Parallel experimental results show that the compound s4 of the present invention inhibits Aurora-B protein kinase activity

The IC50 is 632nM. Example 2 Cancer cell inhibition experiment

According to the standard MTS experiment, the specific parameters are as follows:

Seed plate: 1000 cells / hole

Compound: s4

Cultivation time: 96h

Adding medicine after treatment time: 3h

Compound concentration gradient: Unit: uM Repeat the experiment: 6 groups

Data given form: mean standard deviation

Cell line: human colon cancer cell line (sw620)

The results showed that the compound s4 of the present invention inhibited _SW 620 with an IC50 of about 1 μΜ. See below for details

Example 3

Preparation of pharmaceutical composition (tablet)

The pharmaceutical composition in the form of a tablet is prepared by a conventional technique by mixing the following components and then directly compressing the tablet, and the formulation is as follows:

All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the the In addition, it is to be understood that various modifications and changes may be made by those skilled in the art in the form of the appended claims.

Claims

Rights request

A compound of formula I, 1,2-dihydroxy-3-methyl-9, 10-indole or a pharmaceutically acceptable salt thereof or

2. Use according to claim 1, wherein the medicament is for killing cancer cells or inhibiting the growth of cancer cells.

3. Use according to claim 1 wherein said anticancer drug comprises from 0.001% to 99% by weight of a compound of formula I and a pharmaceutically acceptable carrier or excipient.

4. The use according to claim 1, wherein the medicament is further for inhibiting phosphokinase

Aurora-B.

5. Use according to claim 1, characterized in that the anticancer drug is an injection, granule, capsule, solution or tablet.

6. The use according to claim 1, wherein the cancer is selected from the group consisting of liver cancer, colon cancer, gastric cancer, and lung cancer.

A method for inhibiting growth of tumor cells in vitro, comprising the steps of: adding a compound 1,2-dihydroxy-3-methyl-9, 10-oxime to a culture system of said tumor cells.

The method according to claim 7, wherein the tumor cells are selected from the group consisting of liver cancer cells, colon cancer cells, gastric cancer cells, and lung cancer cells.

A method for treating cancer, comprising the steps of: administering 1,2-dihydroxy-3-methyl-9, 10-oxime or a pharmaceutically acceptable salt thereof to a mammalian subject in need of treatment .

10. The method according to claim 9, wherein the application amount is from 0.1 mg/kg to 20 mg/kg of body weight per day.