阅读说明：本技术 新型联苯衍生物化合物及其用途 (Novel biphenyl derivative compound and use thereof ) 是由 李公珠 李熙允 李济珍 金煌奭 徐知妧 李鸿洙 申智秀 金宝卿 于 2019-05-30 设计创作，主要内容包括：本发明提供了一种新型联苯衍生物化合物或其药学上可接受的盐。本发明的联苯衍生物化合物或其药学上可接受的盐是用于提高Nm23-H1/NDPK活性的物质,并且可以抑制癌症的转移和成长,因此,不仅在预防、改善和治疗癌症方面,而且在抑制癌症转移方面均显示优异的效果。(The present invention provides a novel biphenyl derivative compound or a pharmaceutically acceptable salt thereof. The biphenyl derivative compound or a pharmaceutically acceptable salt thereof of the present invention is a substance for increasing the activity of Nm23-H1/NDPK, and can inhibit the metastasis and growth of cancer, and thus shows excellent effects not only in preventing, ameliorating and treating cancer, but also in inhibiting cancer metastasis.)

1. A biphenyl derivative compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof:

[ formula 1]

Wherein the content of the first and second substances,

l is selected from the group consisting of- (C-C) -, - (NH-C (═ O)) -, - (C (═ O) -NH) -, andany one of the group consisting of, and

R₁to R₄Each independently is hydrogen, hydroxy or C₁To C₃Alkoxy group of (2).

2. The biphenyl derivative compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein C is₁To C₃The alkoxy group of (b) is any one selected from the group consisting of methoxy, ethoxy and propoxy.

3. The biphenyl derivative compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R is₁To R₄Each independently is hydroxy or methylAn oxy group.

4. The biphenyl derivative compound according to claim 3, or a pharmaceutically acceptable salt thereof, wherein R is₁To R₄Each independently is hydroxy or methoxy, and has at least two methoxy groups.

5. The biphenyl derivative compound according to claim 1, wherein L is- (C-C) -, R is₁To R₄Is methoxy.

6. The biphenyl derivative compound according to claim 1, wherein the compound of formula 1 is selected from the group consisting of:

7. a pharmaceutical composition for treating or preventing cancer, comprising the biphenyl derivative compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, as an active ingredient.

8. The pharmaceutical composition of claim 7, wherein the cancer is a cancer selected from the group consisting of: breast cancer, lung cancer, melanoma, prostate cancer, colorectal cancer, bladder cancer, bone cancer, hematological cancer, thyroid cancer, parathyroid cancer, bone marrow cancer, rectal cancer, throat cancer, laryngeal cancer, esophageal cancer, pancreatic cancer, gastric cancer, tongue cancer, skin cancer, brain cancer, uterine cancer, head or neck cancer, gall bladder cancer, oral cancer, colon cancer, anal cancer, central nervous system tumor, liver cancer, colorectal cancer, and combinations thereof.

9. The pharmaceutical composition of claim 7, wherein the cancer is a cancer selected from the group consisting of: breast, lung, colorectal and skin cancers.

10. A method for treating or preventing cancer, the method comprising the step of administering to a subject at risk of or having cancer the pharmaceutical composition of claim 7.

11. A composition for inhibiting cancer metastasis, which comprises the biphenyl derivative compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, as an active ingredient.

12. A method of inhibiting cancer metastasis, the method comprising the step of administering the composition of claim 11 to a subject at risk of or having cancer metastasis.

13. A food composition for preventing or alleviating cancer, comprising the biphenyl derivative compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, as an active ingredient.

Technical Field

The present invention relates to a novel biphenyl derivative compound and use thereof, more particularly, to a novel biphenyl derivative compound; a pharmaceutical composition for treating or preventing cancer, comprising the novel biphenyl derivative compound or a pharmaceutically acceptable salt thereof; a method of treating or preventing cancer comprising the step of administering a pharmaceutical composition; a composition for inhibiting cancer metastasis, which comprises the novel biphenyl derivative compound or a pharmaceutically acceptable salt thereof; a method of inhibiting cancer metastasis comprising the step of administering a pharmaceutical composition; a food composition for preventing or ameliorating cancer, comprising the novel biphenyl derivative compound or a pharmaceutically acceptable salt thereof, and use of the biphenyl derivative compound or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating cancer.

Background

Cancer metastasis (Tumor metastasis Tumor metastasis) is one of the most important factors in determining the prognosis of cancer patients and is also a major cause of cancer-related death. Despite the many efforts made to survive patients by cancer treatments, including surgical, radiation and chemotherapy approaches, there are still efforts to improve the survival of cancer patients. The field of cancer metastasis research is one of the last strategies to overcome cancer, and research on cancer metastasis inhibitors (cancer metastasis inhibitors) is crucial for developing drugs for inhibiting cancer metastasis.

Nm23 is a gene encoding a protein involved in the development and differentiation of normal tissues, and Nm23 has been reported to be reduced in expression in various metastatic cell lines. Generally, Nm23 protein, which consists of 150 to 180 amino acids, contains a leucine zipper motif (leucin zipper motif) and has nucleoside diphosphate kinase (NDPK) activity. In particular, Nm23-H1 has been found to play an important role in cancer metastasis and various other cellular mechanisms, such as cell proliferation, embryonic development, differentiation, and tumor formation. Cancer metastasis occurs through a multi-step process in which cancer cells in primary tumor (primary tumor) tissue first invade blood vessels, then cross blood vessels, survive and form new colonies (colony) at secondary sites. It has been found that Nm23, a nucleoside diphosphate kinase, is a protein that converts NDP (UDP, GDP, and CDP) to NTP (UTP, GTP, and CTP) using ATP, and is an enzyme that regulates the level of intracellular NTP. In addition, overexpression of Nm23-H1 has been found to be closely associated with a reduction in cancer cell invasion. For example, WO1997-035024 discloses a method of treating cancer by administering to cancer cells a combination of enzymes comprising NDPK and nucleoside analogues.

Based on this finding, studies have been directed toward increasing the expression of Nm23 or treatment with Nm23-H1 that is cell permeable (cell permeable). In particular, it has been confirmed that treatment with MPA (Medyxyprogesterone acetate) can increase the expression level of Nm 23-H1. This phenomenon is considered to be a mechanism for inhibiting cancer metastasis by MPA treatment. However, MPA has not been used as a drug since treatment with MPA resulted in an unexpected intracellular response in addition to increasing the level of Nm 23-H1.

In addition, in recent years, methods of inhibiting cancer metastasis using cell-permeable Nm23-H1 have been proposed. The cell-permeable Nm23-H1 is introduced into the cell in the form of a fusion with a transport peptide (transporter peptide) that can cross the cell membrane. As a result, it was confirmed that the treatment with cell-permeable Nm23-H1 showed cancer metastasis inhibitory activity. However, in order to use such cell-permeable Nm23-H1 as a protein drug, its in vivo stability remains a challenge to overcome. In addition, since the drugs for inhibiting cancer metastasis show no significant effect in a short time at the time of treatment, there is a practical problem that expensive drugs such as protein drugs cannot be selected.

Disclosure of Invention

Technical problem

The present inventors have made extensive studies and extensive efforts to develop an agent capable of more effectively inhibiting the development and metastasis of cancer, and as a result, found that the use of a newly developed biphenyl derivative can prevent the development of cancer, treat already developed cancer and inhibit the metastasis of already developed cancer, thereby completing the present invention.

Technical scheme

An object of the present invention is to provide a novel biphenyl derivative compound.

It is another object of the present invention to provide a pharmaceutical composition for treating or preventing cancer, which comprises a novel biphenyl derivative compound or a pharmaceutically acceptable salt thereof.

It is still another object of the present invention to provide a method for treating or preventing cancer, which comprises the step of administering a pharmaceutical composition.

It is still another object of the present invention to provide a pharmaceutical composition for inhibiting cancer metastasis, which comprises a novel biphenyl derivative compound or a pharmaceutically acceptable salt thereof.

It is still another object of the present invention to provide a method for inhibiting cancer metastasis, which comprises the step of administering a pharmaceutical composition.

Still another object of the present invention is to provide a food composition for preventing or ameliorating cancer, which comprises a novel biphenyl derivative compound or a pharmaceutically acceptable salt thereof.

It is still another object of the present invention to provide a biphenyl derivative compound or a pharmaceutically acceptable salt thereof for use in the treatment of cancer.

Advantageous effects

The novel biphenyl derivative compound of formula 1 or a pharmaceutically acceptable salt thereof according to the present invention is a Nm23-H1/NDPK activity enhancing substance, and can inhibit cancer metastasis and growth. Therefore, the composition of the present invention shows excellent effects not only in preventing, ameliorating and treating cancer, but also in inhibiting cancer metastasis.

Brief description of the drawings

Fig. 1 shows the results of evaluating the NDPK activity-enhancing effect of the compound of example 1 (B038) at various concentrations.

Fig. 2 shows the results of evaluating the invasion inhibitory effect of the compound of example 1 (exp.1).

Figure 3 shows the results demonstrating inhibition of cancer metastasis by treatment with the compound of example 1.

Best Mode for Carrying Out The Invention

In order to achieve the above objects, one aspect of the present invention provides a novel biphenyl derivative compound of the following formula 1:

[ formula 1]

Wherein the content of the first and second substances,

l is selected from the group consisting of- (C-C) -, - (NH-C (═ O)) -, - (C (═ O) -NH) -, andany one of the group consisting of; and

R₁to R₄Each independently is hydrogen, hydroxy or C₁To C₃Alkoxy group of (a); wherein C is₁To C₃Is selected from the group consisting of methoxy, ethoxy and propoxyAny one of (1).

Preferably, R in the compound of formula 1₁To R₄Each independently is hydroxy or methoxy. More preferably, R in the compound of formula 1₁To R₄Each independently is hydroxy or methoxy and has at least two methoxy groups.

Further, according to an embodiment of the present invention, the compound of formula 1 is any one selected from the group consisting of:

as used herein, the term "pharmaceutically acceptable salt" refers to salts commonly used in the pharmaceutical art. Examples of salts include: inorganic ion salts formed with calcium, potassium, sodium, magnesium, and the like; inorganic acid salts with hydrochloric acid, nitric acid, phosphoric acid, bromic acid, iodic acid, perchloric acid, sulfuric acid, and the like; organic acid salts formed with acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, and the like; sulfonates formed with methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, and the like; amino acid salts with glycine, arginine, lysine and the like; and amine salts with trimethylamine, triethylamine, ammonia, pyridine, picoline, and the like. However, the type of salt referred to in the present invention is not limited to the above-listed salts.

Another aspect of the present invention provides a method for preparing a biphenyl derivative compound of formula 1.

The biphenyl derivatives can be produced by a continuous or convergent synthetic route through the reaction routes shown in the following reaction schemes 1,2 or 3.

[ reaction scheme 1]

The compound of formula 1 is synthesized by reacting a triphenylphosphine bromide derivative with 2-bromobenzaldehyde, then carrying out a reduction reaction with hydrogen in the presence of platinum oxide, and then reacting with a pinacol borate derivative, potassium carbonate and tetrakis (triphenylphosphine) palladium.

Thus, the compounds of examples 5 to 11 can be prepared.

In the above reaction scheme 1, each R is independently hydrogen or C₁-C₃Alkyl, preferably hydrogen or methyl.

[ reaction scheme 2]

In the above reaction scheme 2, the compound of example 1 can be synthesized by reduction reaction with hydrogen in the presence of Pd/C.

Alternatively, in scheme 2 above, the compound of example 2 can be prepared by reacting NaHCO₃The aqueous solution was added to 2- (3, 4-dimethoxystyryl) -3',4' -dimethoxybiphenyl and reacted to synthesize the compound.

In the above reaction scheme 2, R₁To R₄As defined in formula 1, and L₁Is- (C-C) -or

[ reaction scheme 3]

In the above reaction scheme 3, the compounds of examples 3 and 4 can be synthesized by reacting the above boronic acid derivatives with reactants after adding a Pd catalyst and potassium carbonate (3.0 equivalents) to dioxane and distilled water.

In the above reaction scheme 3, R₁To R₄As defined in formula 1, and L₂Is- (NH-C (═ O)) -or- (C (═ O))-NH)-。

According to one embodiment of the present invention, the biphenyl derivative compound of formula 1 may be synthesized according to the sequence shown in the reaction scheme, but may also be synthesized by the method proposed herein or a similar method. Thus, the synthetic route thereof is not limited to the route shown in the reaction scheme. The starting materials are commercially available or can be prepared by methods similar to those shown below.

The separation and purification of the biphenyl derivative compound or the intermediate synthesized by the above method may be performed by any suitable separation or purification method used in the pharmaceutical field, such as filtration, extraction, crystallization, column chromatography, thin layer chromatography, thick layer chromatography, preparative low pressure or high pressure liquid chromatography, or a combination of these methods.

Still another aspect of the present invention provides a pharmaceutical composition for treating or preventing cancer, comprising a biphenyl derivative compound of formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In the present invention, the cancer is not particularly limited as long as it can be treated or prevented by the biphenyl derivative compound of formula 1 or a pharmaceutically acceptable salt thereof provided according to the present invention. In one example, the cancer can be breast cancer, lung cancer, melanoma, prostate cancer, colorectal cancer, bladder cancer, bone cancer, hematological cancer, thyroid cancer, parathyroid cancer, bone marrow cancer, rectal cancer, throat cancer, larynx cancer, esophageal cancer, pancreatic cancer, gastric cancer, tongue cancer, skin cancer, brain tumor, uterine cancer, head or neck cancer, gall bladder cancer, oral cancer, colon cancer, anal cancer, central nervous system tumor, liver cancer, large intestine cancer, and the like. In another example, the cancer can be breast cancer, lung cancer, colorectal cancer, skin cancer, and the like.

According to one embodiment of the present invention, the biphenyl derivative compound of formula 1 or a pharmaceutically acceptable salt thereof is a substance enhancing the activity of Nm23-H1/NDPK, and can inhibit cancer metastasis and growth.

Accordingly, the biphenyl derivative compound of formula 1 or a pharmaceutically acceptable salt thereof shows an effect not only in the prevention or treatment of cancer but also in the inhibition of cancer metastasis.

As used herein, the term "treatment" refers to any activity that ameliorates or beneficially alters the symptoms of cancer by the administration of a biphenyl derivative compound or a pharmaceutically acceptable salt thereof.

As used herein, the term "preventing" refers to inhibiting or delaying any activity of cancer by administering a biphenyl derivative compound or a pharmaceutically acceptable salt thereof.

The pharmaceutical composition of the present invention may include 0.001 to 80 wt%, specifically 0.001 to 70 wt%, more specifically 0.001 to 60 wt% of the biphenyl derivative compound or the pharmaceutically acceptable salt thereof based on the total weight of the composition, but is not limited thereto.

For administration, the pharmaceutical composition of the present invention may further comprise at least one pharmaceutically acceptable carrier in addition to the biphenyl derivative compound of formula 1 or a pharmaceutically acceptable salt thereof. As pharmaceutically acceptable carriers, saline, sterile water, ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, or a mixture of two or more thereof may be used. The pharmaceutical composition may contain other conventional additives, such as antioxidants, buffers and bacteriostats, if necessary. In addition, the pharmaceutical composition may be formulated into an injectable preparation, such as an aqueous solution, suspension or emulsion, or a pill, capsule, granule or tablet by further adding a diluent, a dispersant, a surfactant, a binder and a lubricant. Thus, the pharmaceutical composition of the present invention may be in the form of a patch, liquid, pill, capsule, granule, tablet, suppository, etc. These preparations can be prepared by any conventional preparation method for preparations used in the art or a method disclosed in Remington's Pharmaceutical Science (latest edition), Mack publishing company, Iston Pa, and can be prepared in various forms depending on each disease condition or its components.

The pharmaceutical composition of the present invention can be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally or topically) according to a desired method, and its administration dose can be in a wide range depending on the body weight, age, sex, health condition and diet of a patient, administration duration, administration mode, excretion rate and severity of disease. The compounds of formula 1 according to the invention can be administered once or several times daily in a daily dose of about 1 to 1000mg/kg, preferably 5 to 100 mg/kg.

The pharmaceutical composition of the present invention may further comprise at least one active ingredient exhibiting the same or similar pharmaceutical effect as that of the biphenyl derivative compound of formula 1 or a pharmaceutically acceptable salt thereof, in addition to the biphenyl derivative compound of formula 1 or a pharmaceutically acceptable salt thereof.

The present invention also provides a method for treating or preventing cancer, the method comprising the step of administering to a subject at risk of or having cancer a biphenyl derivative compound, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition for treating or preventing cancer.

The terms "biphenyl derivative compound", "pharmaceutically acceptable salt", "treating" and "preventing" are defined herein above.

As used herein, the term "subject" refers to all animals, including humans, rats, mice, and livestock, that have or are at risk of developing cancer. In a particular example, the subject may be a mammal, including a human.

The pharmaceutical compositions of the present invention are administered in a therapeutically effective amount. The term "therapeutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to any medical treatment. An effective dosage level of a pharmaceutical composition may be determined based on factors including the type of subject, the severity of the disease, the age and sex, the activity of the drug, the sensitivity to the drug, the duration of administration, the route of administration, the rate of excretion, the duration of treatment, the drug used in combination with the composition, and other factors well known in the medical arts. For example, the biphenyl derivative compound or a pharmaceutically acceptable salt thereof may be administered at a daily dose of 0.01 to 500mg/kg, specifically at a daily dose of 10 to 100mg/kg, and the administration may be performed one or more times per day.

The treatment methods of the present invention also include inhibiting or avoiding the symptoms of the disease prior to the onset of symptoms and addressing the disease itself by administering a compound of formula 1. In the management of diseases or conditions, the prophylactic or therapeutic dose of a particular active ingredient may vary according to the nature (nature) and severity of the disease or condition and the route of administration of the active ingredient. The dose and frequency of administration of the dose will also vary according to the age, weight and response of the individual patient. Taking these factors into account, one skilled in the art can readily select an appropriate dosage regimen. In addition, the treatment methods of the present invention may further comprise administering a therapeutically effective amount of an additional active agent useful for treating the disease with the compound of formula 1, wherein the additional active agent may exhibit a synergistic or adjunctive effect with the compound of formula 1.

The present invention also provides a pharmaceutical composition for inhibiting cancer metastasis, which comprises a biphenyl derivative compound of formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. The present invention also provides a method for inhibiting cancer metastasis, the method including the step of administering a composition for inhibiting cancer metastasis to a subject at risk of cancer metastasis or who has cancer metastasis.

The terms "biphenyl derivative compound", "pharmaceutically acceptable salt", "treating", "preventing", and "subject" are defined herein as above.

The composition of the present invention shows an excellent effect of inhibiting cancer metastasis by increasing Nm23-H1/NDPK activity.

The present invention also provides a food composition for preventing or alleviating cancer, which comprises a biphenyl derivative compound of formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

The food composition of the present invention can be used as a health functional food. The term "health functional food" means a food which is produced and processed using raw materials or ingredients which are beneficial to the human body's functionality and meet the regulation of "health functional food act" No. 6727. The term "functional" refers to the ingestion of food intended to control the structural and functional nutrition of the human body or to produce beneficial effects on health, such as physiological effects.

The food composition of the present invention may comprise conventional food additives. Unless otherwise stated, the applicability of the food additive should be determined by the specifications and standards in the relevant provisions of the korean food additive code and general test methods, which are approved by the korean food and drug administration.

The food composition of the present invention may comprise 0.01 to 95 wt%, preferably 1 to 80 wt%, of the compound of formula 1 based on the total weight of the composition for the purpose of preventing and/or ameliorating cancer. In addition, the food composition may be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills, beverages, etc. for the purpose of preventing and/or improving cancer.

The present invention also provides the use of a biphenyl derivative compound of formula 1 or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of cancer.

The compound of formula 1 used for preparing a medicament may be mixed with an acceptable adjuvant, diluent or carrier, etc., and may be combined with other active ingredients to form a combined preparation exhibiting a synergistic effect between the active ingredients.

The matters mentioned in the compositions, uses and methods of treatment of the present invention apply equally unless they contradict each other.

EMBODIMENTS FOR CARRYING OUT THE INVENTION

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily practice the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

In the following examples of the invention, reagents and solvents mentioned below were purchased from Sigma-Aldrich (Sigma-Aldrich), TCI, and HPLC was performed using a chiral IB chromatography column (Hex/iPA ═ 80/20, 0.5mL/min), unless otherwise indicated. Silica gel 60(230-400 mesh ASTM) was used as the silica gel for column chromatography. AV300 (30) using the Brookfield Fourier transform0MHz) spectrometer, Brookfield Fourier transform AV400(400MHz) spectrometer or Agilent technologies DD2(600MHz) detection¹H NMR data.

EXAMPLE 1.2- (3, 4-Dimethoxyphenethyl) -3',4' -dimethoxy-1, 1' -biphenyl (HYL-NM-038) Synthesis of (2)

O-bromobenzaldehyde (1.0 eq) was dissolved in DMF and 3, 4-dimethoxyphenylboronic acid (1.0 eq), 2M Na₂CO₃Aqueous solution (3.0 equiv.) and Pd (PPh)₃)₄(0.01 eq) was added thereto at room temperature. The mixture was stirred at 80 ℃ overnight and then diluted with ethyl acetate. Then, an aqueous ammonium chloride solution and water were added thereto to terminate the reaction. Next, the reaction mixture was extracted with ethyl acetate, and the organic layer was then MgSO₄And (5) drying. The mixture was filtered, and the collected organic layer was concentrated and then purified on silica gel using column chromatography, thereby synthesizing the target compound.

¹H NMR(300MHz,CDCl₃)δ9.98(s,1H),7.98(d,J＝7.9Hz,1H),7.61(td,J＝7.5,1.4Hz,1H),7.45(t,J＝8.0Hz,2H),6.98-6.85(m,3H),3.93(s,3H),3.89(s,3H).

The compound synthesized above was added to a solution of dialkyl benzylphosphonate (2.0 equivalents) in tetrahydrofuran at 0 ℃ along with n-BuLi (2.48M in n-hexane) (1.95 equivalents). The mixture was stirred at room temperature for 30 minutes, then cooled to 0 ℃ and a solution of cis-aldehyde (1.0 equiv.) and DMPU (5.0 equiv.) in THF was added thereto using a cannula. The resulting mixture was stirred at room temperature overnight, and then the reaction was terminated by adding an aqueous ammonium chloride solution and water thereto. The reaction mixture was extracted with ethyl acetate and MgSO₄Dried, concentrated and then purified on silica gel using column chromatography.

¹H NMR(599MHz,CDCl₃)δ7.70(d,J＝7.6Hz,1H),7.33(d,J＝7.1Hz,2H),7.30(d,J＝6.2Hz,1H),7.03-6.91(m,6H),6.89(d,J＝1.6Hz,1H),6.80(d,J＝8.3Hz,1H),3.92(s,3H),3.86(s,3H),3.84(s,3H),3.83(s,3H).

The compound synthesized above was dissolved in a 1:1 solvent of methanol and dichloromethane. After Pd/C (10 wt%) (0.1 equivalent) was added thereto, the mixture was stirred under a hydrogen atmosphere for 3 hours. Thereafter, the stirred mixture was filtered through celite and silica, the organic solvent was evaporated, and then the concentrated organic compound was purified on silica gel using column chromatography to give the desired product [2- (3, 4-dimethoxyphenethyl) -3',4' -dimethoxy-1, 1' -biphenyl ].

¹H NMR (300MHz, chloroform-d) δ 7.31-7.25(m,2H),7.24-7.17(m,2H),6.89(d, J ═ 8.6Hz,1H),6.79(dt, J ═ 4.3,2.2Hz,2H),6.68(d, J ═ 8.1Hz,1H),6.49(dd, J ═ 8.1,2.0Hz,1H),6.35(d, J ═ 2.0Hz,1H),3.91(s,3H),3.84(s,3H),3.81(s,3H),3.73(s,3H),2.95-2.81(m,2H),2.73-2.60(m,2H).

EXAMPLE 2- (3',4' - -dimethoxy- [1,1' -biphenyl)]-2-yl) -3- (3, 4-dimethoxyphenyl) ring Synthesis of ethylene oxide (HYL-NM-039)

NaHCO is added₃The aqueous solution was added to a solution of 2- (3, 4-dimethoxystyryl) -3',4' -dimethoxybiphenyl (1.0 eq) in acetone and acetonitrile and cooled to a temperature of 0 ℃. Potassium peroxymonosulfonate (2.2 equivalents) was added thereto, followed by stirring for 4 hours. Distilled water was added thereto to terminate the reaction, and then the reaction mixture was extracted with ethyl acetate and MgSO₄Dried and then concentrated. The resulting mixture was purified using column chromatography on silica gel. To obtainThe desired product [2- (3',4' -dimethoxy- [1,1' -biphenyl ]]-2-yl) -3- (3, 4-dimethoxyphenyl) oxirane]。

¹H NMR (300MHz, benzene-d 6) δ 7.84-7.76(m,1H),7.50-7.37(m,3H),7.27(dd, J ═ 7.1,2.1Hz,2H),7.07-6.85(m,4H),6.61(d, J ═ 8.2Hz,1H),6.50(d, J ═ 8.2Hz,1H),3.40(s,3H),3.36(d, J ═ 1.1Hz,6H),3.33(s,3H).

Example 3N- (3, 4-Dimethoxyphenyl) -3',4' -dimethoxy- [1,1' -biphenylyl ] benzene]-2-carboxamides Synthesis of (HYL-NM-041)

2ml of DMF was added to a solution of 2-iodobenzoic acid (1.0 eq) in dichloromethane at 0 ℃. Then, oxalyl chloride (2.0 equivalents) was added dropwise thereto, followed by stirring for 1 hour. Next, 3, 4-dimethoxyaniline was added to the mixture, which was then warmed to room temperature and stirred for 2 and a half hours. The reaction mixture was diluted with dichloromethane, and then water was added thereto to terminate the reaction. The mixture was extracted with dichloromethane and MgSO₄Dried and then concentrated under vacuum. Next, the concentrate was purified on silica gel using column chromatography to give the desired product.

Tetrakis (triphenylphosphine) palladium (0.01 eq) and K were added at room temperature₂CO₃(3.0 equiv.) to a solution of N- (3, 4-dimethoxyphenyl) -2-iodobenzamide (1.0 equiv.) and 3, 4-dimethoxyphenylboronic acid (2.0 equiv.) in DMF (1ml) and distilled water (0.1 ml). Then, the mixture was stirred at 110 ℃ for 16 hours. Next, an aqueous ammonium chloride solution was added thereto to terminate the reaction, and then the reaction mixture was extracted with ethyl acetate, MgSO₄Dried and then concentrated under vacuum. The concentrate was then purified by column chromatography on silica gel to give the desired product [ N- (3, 4-dimethoxyphenyl)) -3',4' -dimethoxy- [1,1' -biphenyl]-2-carboxamides]。

¹H NMR (300MHz, chloroform-d) δ 8.02(s,1H),7.88(dd, J ═ 7.5,1.6Hz,1H),7.61-7.40(m,3H),7.07(dd, J ═ 8.2,2.1Hz,1H),7.03-6.93(m,4H),6.73(d, J ═ 8.6Hz,1H),6.52(dd, J ═ 8.6,2.5Hz,1H),3.93(s,3H),3.84(d, J ═ 2.2Hz,6H),3.81(s,3H),2.97(s,1H),2.90(s,1H).

Example 4N- (3',4' -dimethoxy- [1,1' -biphenyl)]-2-yl) -3, 4-dimethoxybenzamide (HYL- Synthesis of NM-042)

To a solution of 3, 4-dimethoxybenzoic acid (1.0 eq.) in dichloromethane was added DMF at 0 ℃ in the amount of 1/10 parts of dichloromethane. Oxalyl chloride (2.0 equivalents) was added dropwise thereto, followed by stirring for 1 hour. The mixture was warmed to room temperature and then stirred for further 30 minutes. Thereafter, 2-iodoaniline (5.0 equivalents) was added thereto, followed by further stirring for 2 hours. 5mL of tetrahydrofuran was further added to the mixture, followed by further stirring for 30 minutes, and then distilled water was added thereto to terminate the reaction. The mixture was extracted with dichloromethane and MgSO₄Dried and then concentrated under vacuum. The concentrate was purified using column chromatography on silica gel to give the desired product.

¹H NMR (300MHz, chloroform-d) δ 8.41(dd, J ═ 8.3,1.6Hz,1H),8.23(s,1H),7.78(dd, J ═ 8.0,1.5Hz,1H),7.61-7.44(m,2H),7.36(ddd, J ═ 8.6,7.4,1.5Hz,1H),6.93(d, J ═ 8.2Hz,1H),6.88-6.79(m,1H),3.94(s,4H),3.93(s,3H).

N- (2-iodophenyl) -3, 4-dimethoxybenzamide (1.0 eq), 3, 4-dimethoxyboronic acid (2.0 eq), Pd catalyst (0.01 eq) and potassium carbonate (3.0 eq) were dissolved in dioxane and distilled water (10: 1). The mixture was stirred at 100 ℃ 1After 3 hours, an aqueous ammonium chloride solution was added thereto to terminate the reaction. The organic mixture was extracted with ethyl acetate, MgSO₄Dried and then concentrated under vacuum. The concentrate was then purified by column chromatography on silica gel to give the desired product [ N- (3',4' -dimethoxy- [1,1' -biphenyl ]]-2-yl) -3, 4-dimethoxybenzamide]。

¹H NMR (300MHz, chloroform-d) δ 8.50(dd, J ═ 8.1,1.2Hz,1H),8.04(s,1H),7.39(ddd, J ═ 8.3,7.2,1.7Hz,1H),7.32 to 7.22(m,2H),7.16(td, J ═ 7.5,1.3Hz,1H),7.06 to 6.95(m,3H),6.94 to 6.88(m,1H),6.78(d, J ═ 8.4Hz,1H),3.92(s,3H),3.87(s,3H),3.84(s,3H),3.81(s,3H).

Examples 5 to 11 Synthesis of Compounds (HYL-NM-049 to 055)

General procedure A

The triphenylphosphine bromide derivative (1.2 equivalents) was added to tetrahydrofuran at 0 ℃ and t-BuOK (1.4 equivalents) was added thereto. After 30 minutes, a solution of 2-bromobenzaldehyde (1.0 eq) in tetrahydrofuran was added to the mixture over 10 minutes. The reaction mixture was warmed to room temperature and then stirred for 16 hours. Then, distilled water was added thereto to terminate the reaction, and the reaction mixture was extracted with ethyl acetate. The extract was MgSO₄Drying, concentration, and purification by column chromatography on silica gel gave the desired product.

The synthesized bromide (1 equivalent) was dissolved in dichloromethane, and then platinum oxide (0.73 equivalent) was added thereto under a hydrogen atmosphere. The mixture was stirred for 6 hours, filtered through silica and celite, and then concentrated under vacuum. The concentrate was purified using column chromatography on silica gel to give the desired product.

General procedure B

Borane pinacol ester derivative (1.5 equiv.), potassium carbonate (3.0 equiv.) and tetrakis (triphenylphosphine) palladium (0.05 equiv.) were dissolved in DME/distilled water (2/1). To this mixture was slowly added dropwise a solution of bromide (1.0 equiv) in DME using a cannula. The mixture was mixed at 90 ℃ until complete disappearance of the starting material. After completion of the reaction, the reaction mixture was cooled to room temperature, and then distilled water was added to the mixture to terminate the reaction. The reaction mixture was extracted with ethyl acetate and washed with brine. The organic mixture was over MgSO₄Drying, concentration, and purification by column chromatography on silica gel gave the desired product.

General procedure C (Synthesis of reactants in general procedure A)

A solution of benzyl alcohol derivative (1.0 equiv.) and triphenylphosphine hydrogen bromide (1.0 equiv.) in acetonitrile was stirred at 85 ℃ for 2 h. When the product was synthesized as a salt, it was cooled to room temperature and then filtered. The product was washed with acetonitrile.

The following seven compounds were synthesized according to the general procedures a, B and C using reactants in which R at each position was correctly substituted with H or Me.

EXAMPLE 5 '- (3, 4-Dimethoxyphenethyl) -3-methoxy- [1,1' -biphenyl]-4-alcohols

¹H NMR (400MHz, chloroform-d) δ 7.25(d, J ═ 2.5Hz,1H),7.24-7.15(m,2H),6.86(dd, J ═ 5.0,3.1Hz,2H),6.77-6.65(m,2H),6.53(dd, J ═ 8.1,1.9Hz,1H),6.40(d,J＝2.0Hz,1H),3.91(s,3H),3.81(d,J＝1.1Hz,3H),3.74(s,3H),2.93-2.82(m,2H),2.71-2.61(m,2H).

EXAMPLE 6 '- (3, 4-Dimethoxyphenethyl) -4-methoxy- [1,1' -biphenyl]-3-alcohols

¹H NMR (400MHz, chloroform-d) δ 7.35-7.25(m,2H),7.22-7.15(m,2H),6.89-6.81(m,2H),6.75-6.65(m,2H),6.53(dd, J ═ 8.1,2.0Hz,1H),6.40(d, J ═ 2.0Hz,1H),5.63(s,1H),3.91(s,3H),3.81(s,3H),3.74(s,3H),2.92-2.82(m,2H),2.69-2.59(m,2H).

EXAMPLE 7.5- (2- (3',4' -dimethoxy- [1,1' -biphenyl)]-2-yl) ethyl) -2-methoxyphenol

In general procedure B, boronic acid was used instead of pinacol boronate. DME and H used as solvent₂The ratio between O is from 2:1 is changed into 5: 1.

¹h NMR (400MHz, chloroform-d) δ 7.29-7.25(m,2H),7.23-7.18(m,1H),6.90(d, J ═ 8.1Hz,1H),6.85-6.77(m,2H),6.69(dd, J ═ 15.1,8.4Hz,1H),6.57(d, J ═ 2.1Hz,1H),6.51-6.39(m,1H),3.92(s,3H),3.84(s,3H),3.82(s,3H),2.96-2.73(m,2H),2.69-2.54(m,2H).

EXAMPLE 8 '- (3-hydroxy-4-methoxyphenethyl) -3-methoxy- [1,1' -biphenyl]-4-alcohols

¹H NMR (400MHz, chloroform-d) δ 7.29-7.23(m,2H),7.22-7.16(m,2H),6.90-6.83(m,2H),6.76(dd, J ═ 8.2,2.1Hz,1H),6.68(d, J ═ 8.1Hz,1H),6.58(d, J ═ 2.1Hz,1H),6.48(dd, J ═ 8.2,2.1Hz,1H),5.65(s,1H),5.52(s,1H),3.93(s,3H),3.82(s,3H),2.92-2.78(m,2H),2.72-2.57(m,2H).

Example 9 '- (3-hydroxy-4-methoxyphenethyl) -4-methoxy- [1,1' -biphenyl]-3-alcohols

¹H NMR (400MHz, chloroform-d) δ 7.27-7.24(m,2H),7.23-7.15(m,2H),6.91-6.84(m,2H),6.76(dd, J ═ 8.2,2.1Hz,1H),6.68(d, J ═ 8.2Hz,1H),6.58(d, J ═ 2.1Hz,1H),6.48(dd, J ═ 8.2,2.1Hz,1H),5.65(s,1H),5.52(s,1H),3.92(s,3H),3.82(s,3H),2.96-2.74(m,2H),2.74-2.58(m,2H).

EXAMPLE 10.4- (2- (3',4' -dimethoxy- [1,1' -biphenyl)]-2-yl) ethyl) -2-methoxyphenol

In general procedure B, boronic acid was used instead of pinacol boronate. DME and H used as solvent₂The ratio between O was changed from 2:1 to 5: 1.

¹H NMR (400MHz, chloroform-d) δ 7.36-7.25(m,2H),7.24-7.14(m,2H),6.93-6.85(m,1H),6.82-6.75(m,2H),6.72(d, J ═ 8.0Hz,1H),6.44(dd, J ═ 8.0,1.9Hz,1H),6.31(d, J ═ 1.9Hz,1H),5.40(s,1H),3.90(s,3H),3.84(s,3H),3.73(s,3H),2.90-2.79(m,2H),2.70-2.55(m,2H).

EXAMPLE 11 '- (4-hydroxy-3-methoxyphenethyl) -3-methoxy- [1,1' -biphenyl]-4-alcohols

¹H NMR (400MHz, chloroform-d) δ 7.26-7.24(m,2H),7.23-7.13(m,2H),6.89-6.82(m,2H),6.78-6.62(m,2H),6.48(dd, J ═ 8.0,2.0Hz,1H),6.38(d, J ═ 1.9Hz,1H),5.66(s,1H),5.44(s,1H),3.91(s,3H),3.75(s,3H),2.91-2.79(m,2H),2.71-2.57(m,2H).

The structures of the compounds of the present invention synthesized in examples 1 to 11 are summarized in table 1 below.

[ Table 1]

Example 12 validation of enhanced NDPK Activity

5ng of recombinant Nm23-H1 with each test compound (examples 1 to 3) in NDPK assay buffer (20mM HEPES, 3mM MgCl) at room temperature₂) Incubated for 10 min and then reacted with 5 μ M ADP for 1 min for NDPK assay.

The same effect was confirmed by cell-based NDPK assay, which was performed as follows: 5,000K MDA-MB-231 cells were lysed with protease inhibitor cocktail (protease inhibitor cocktail) and NDPK assay buffer (NDPK assay buffer), and the resulting cell lysates were centrifuged at 8,000rpm for 10 min at 4 ℃. mu.L of the lysate was incubated with each test compound for 5 minutes, then 50. mu.M UDP was added thereto, and then reacted with NDPK. ATP consumption was assessed by ATP assay kit (molecular probe, usa).

The results are shown in table 2 below.

[ Table 2]

EXAMPLES Compounds	NDPK Activity
		1	6.55
2	4.08
		3	1.32
4	1.52
		5	4
6	1.9
		7	3.5
10	2.6

As described above, it was confirmed that the example compounds exhibited excellent effects of enhancing NDPK activity (enhancing Nm23-H1 activity).

Example 13 verification of the NDPK Activity-enhancing Effect of the Compound of example 1 at various concentrations

The NDPK activity-enhancing effect of the compound of example 1 at various concentrations was evaluated in the same manner as in example 2, and the results are shown in fig. 1.

As shown in fig. 1, it was confirmed that the treatment with the compound of example 1 showed an excellent effect of enhancing NDPK activity even at a low concentration.

Example 14 verification of cancer metastasis inhibition by matrigel invasion assay

Having polycarbonate membranes (pore size: 8 μm) with or without a matrigel coatingUnit cell (Corning, USA) for invasionAnd (4) measuring. The membrane was coated with 50. mu.g of Matrigel^TM-membrane matrix of the base (BD biosciences, usa). Will be 5X 10⁴Individual MDA-MB-231 cells were seeded into the upper chamber in serum-free medium with the compound of example 1 and the lower chamber was filled with medium containing 10% FBS. After incubation at 37 ℃ for 24 hours, the cells on the upper side of the membrane were removed. Cells that invade the underside of the membrane were stained with 0.5% crystal violet in 25% methanol and counted under a microscope at 100 x magnification.

The results are shown in FIG. 2.

Fig. 2(a) shows a photomicrograph, and fig. 2(B) shows the results of counting the invaded cells. As shown in fig. 2, it was confirmed that the treatment with the compound of example 1 significantly inhibited cell invasion.

Example 15 verification of cancer metastasis inhibition by immunofluorescence staining assay

MDA-MB-231 cells in Secureslip^TM(sigma) cell culture glass coverslips were grown to a confluency of 50-70% and then treated multiple times with or without the compound of example 1 (B038). Cells were gently washed with cold HBSS and then fixed with HBSS with 4% paraformaldehyde in RBS for 10 min at room temperature. After washing with HBSS, permeabilization with 0.1% Triton X-100(Triton-X) for 10 minutes at room temperature. After washing twice with HBSS, cells were blocked with 3% BSA, 0.2% tween 20 and 0.2% gelatin in HBSS for 1 hour at room temperature and incubated with primary antibody for 2 hours at 37 ℃. F-actin was stained with rhodamine-phalloidin (Saimer fly technology) for 2 hours at 37 ℃ and then washed 3 times with HBSS for 20 minutes. Coverslips (Cover slides) were treated with an anti-fade solution (anti-fading solution) and observed using the x63 objective of an LSM510 META laser scanning confocal microscope.

The results are shown in FIG. 3.

As shown in FIG. 3, it was confirmed from the localization of F-actin in MDA-MB-231 cells that wrinkles (Ruffle) were reduced and cell-cell contact was increased (B038). This indicates that the compound of example 1 inhibits cancer cell metastasis by inhibiting the activity of Rac1 involved in cell migration through Nm23-H1 and reducing cell migration.

In combination with the above results, the novel biphenyl derivative compounds of the present invention inhibit cancer metastasis by enhancing NDPK activity. That is, the novel compound of the present invention can exhibit an excellent effect of inhibiting cancer metastasis, and thus can be used as an anticancer agent and an anticancer adjuvant exhibiting excellent prophylactic and therapeutic effects.

Although the present invention has been described in detail with reference to the specific features, it is apparent to those skilled in the art that such detailed description is only a preferred embodiment thereof, and does not limit the scope of the present invention. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.