JP2011016754A - Pharmaceutical composition for preventing and/or treating cancer comprising substituted phenanthrene compound as active ingredient - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a compound effective for preventing and/or treating cancer.SOLUTION: The pharmaceutical composition for preventing and/or treating cancer comprises a substituted phenanthrene compound, a salt thereof, a solvate thereof or a physiologically functional derivative thereof as an active ingredient. In the substituted phenanthrene compound, phenanthrene is substituted by: 1-9 substituents selected from the group consisting of a hydroxy group, halogen, 1C-6C alkoxy group and 1C-6C alkoxy-1C-6C alkyl group; and 1-3 hydrophilic groups selected from the group consisting of a carboxy group, sulfonic acid group, hydroxy group, hydroxy 1C-6C alkyl group, amide group, ethylene oxide group, propylene oxide group and ammonium group.

Description

  The present invention relates to a pharmaceutical composition for preventing and / or treating cancer comprising a substituted phenanthrene compound as an active ingredient.

  With the progress of elucidation of the molecular mechanism of the carcinogenic mechanism, development of so-called molecular target drugs targeting molecules that are closely involved in the carcinogenic mechanism, in particular, signal transduction molecules, has been actively carried out in recent years. Signal transduction molecules that can be targets of molecular target drugs are rarely expressed in normal cells, but are highly expressed in cancer cells and play an essential role in the growth process of cancer cells. Plays. The molecularly targeted drugs that have been developed so far target the growth factor receptor itself that satisfies these conditions or a tyrosine kinase located downstream thereof.

  Our group analyzed the gene expression pattern in a mouse liver cancer development model, and Pim-, a proto-oncogene having serine / threonine kinase activity that is not observed in normal liver tissue. It has been clarified that the expression of 3 is enhanced at the stage after the precancerous lesion in the liver of the mouse, and the same enhanced expression of Pim-3 is also observed in the human liver cancer lesion (Non-patent Document 1). ). Furthermore, suppression of Pim-3 expression in human hepatoma cell lines with RNAi delays cell cycle progression with increased apoptosis, indicating that Pim-3 is a cell growth process in hepatoma cell lines. I came to assume the possibility of being involved in. At this time, the entire nucleotide sequence of human Pim-3 cDNA, which had not been determined so far, was also determined.

  In the human pancreas, large intestine, and stomach, which are organs derived from the endoderm as well as in the liver, even though Pim-3 protein is not detected normally, about 50% of cases are colon cancer, gastric cancer, and liver cancer. In all cases of pancreatic cancer, Pim-3 protein expression is up-regulated, and even in these cancer cell lines, suppression of Pim-3 expression delays cell cycle progression with increased apoptosis It was also found (Non-Patent Documents 2 and 3). These results suggest that Pim-3 is also involved in the carcinogenesis process of endoderm-derived organ cancers, especially pancreatic cancer, and is a target molecule for the treatment of these cancers. It has also been clarified that Pim-3 suppresses apoptosis by selectively phosphorylating and inactivating the 112th serine residue of Bad, which is an apoptotic molecule (Non-patent Document 2, 3).

  The existence of Pim-1 and Pim-2 has been reported in the Pim family, and both have been reported to phosphorylate the 112th serine residue of Bad. Furthermore, Pim-1 shows high homology with Pim-3 including the active center, which has been revealed from the analysis of the entire nucleotide sequence of cDNA. However, Pim-1 has been reported to be a malignant tumor of hematopoietic organs, and Pim-2 has been reported to be upregulated in prostate cancer. However, in endoderm-derived organ cancers, especially pancreatic cancer, Pim-1 and Pim- There is no report that the expression of 2 is increased.

Int. J. Cancer 114: 209-218, 2005 Cancer Res. 66: 6741-6747, 2006 Cancer Sci. 98: 321-328, 2007

  An object of the present invention is to provide a compound effective for the prevention and / or treatment of cancer.

  The present inventors examined the influence of candidate compounds on Pim-1, Pim-2 and Pim-3 activities by enzyme immunological methods. As a result, they found compounds that significantly inhibited these activities, and Was found to suppress the proliferation of cancer cell lines in vitro, and the present invention was completed.

[式中、
R¹およびR²は、独立に、ヒドロキシ基、ハロゲン、C_1-6アルコキシ基、およびC_1-6アルコキシ-C_1-6アルキル基からなる群から選択される置換基であり、
R³は、カルボキシ基、スルホン酸基、ヒドロキシ基、ヒドロキシC_1-6アルキル基、アミド基、エチレンオキシド基、プロピレンオキシド基、およびアンモニウム基からなる群から選択される親水基であり、
nおよびmは、独立に、0〜4であり、ただし、n + mは1〜8であり、
pは、1または2である]
で表される化合物である、（１）記載の医薬組成物。
（３）置換フェナントレン化合物が、式II：

[式中、
R¹、R²およびR³は、（２）で定義したとおりであり、
nおよびmは、独立に、1または2であり、ただし、n + mは3である]
で表される化合物である、（２）記載の医薬組成物。
（４）置換フェナントレン化合物が、式IIa、IIbまたはIIc：

[式中、R¹、R²およびR³は、（２）で定義したとおりである］
で表される化合物である、（３）記載の医薬組成物。
（５）R¹およびR²が、独立に、C_1-6アルコキシ基であり、R³がカルボキシ基である、（２）〜（４）のいずれかに記載の医薬組成物。
（６）置換フェナントレン化合物が、以下：

からなる群から選択される化合物である、（４）記載の医薬組成物。
（７）がんは、膵臓がん、胃がん、大腸がん、腎臓がん、肝臓がん、骨髄がん、副腎がん、皮膚がん、メラノーマ、肺がん、小腸がん、前立腺がん、精巣がん、子宮がん、乳がんまたは卵巣がんである、（１）〜（６）のいずれかに記載の医薬組成物。
（８）がんは膵臓がんである、（７）記載の医薬組成物。 That is, the present invention includes the following inventions.
(1) The phenanthrene is 1 to 9 substituents selected from the group consisting of a hydroxy group, a halogen, a C _1-6 alkoxy group, and a C _1-6 alkoxy-C _1-6 alkyl group, and a carboxy group, Substituted phenanthrene compounds substituted with 1 to 3 hydrophilic groups selected from the group consisting of sulfonic acid groups, hydroxy groups, hydroxy C _1-6 alkyl groups, amide groups, ethylene oxide groups, propylene oxide groups, and ammonium groups Or a salt, solvate, or physiologically functional derivative thereof as an active ingredient for a pharmaceutical composition for preventing and / or treating cancer.
(2) Substituted phenanthrene compounds have the formula I:

[Where
R ¹ and R ² are independently a substituent selected from the group consisting of a hydroxy group, a halogen, a C _1-6 alkoxy group, and a C _1-6 alkoxy-C _1-6 alkyl group;
R ³ is a hydrophilic group selected from the group consisting of a carboxy group, a sulfonic acid group, a hydroxy group, a hydroxy C _1-6 alkyl group, an amide group, an ethylene oxide group, a propylene oxide group, and an ammonium group;
n and m are independently 0-4, provided that n + m is 1-8,
p is 1 or 2]
The pharmaceutical composition according to (1), which is a compound represented by the formula:
(3) Substituted phenanthrene compounds have the formula II:

[Where
R ¹ , R ² and R ³ are as defined in (2),
n and m are independently 1 or 2, where n + m is 3]
The pharmaceutical composition according to (2), which is a compound represented by:
(4) the substituted phenanthrene compound is of formula IIa, IIb or IIc:

[Wherein R ¹ , R ² and R ³ are as defined in (2)]
The pharmaceutical composition according to (3), which is a compound represented by:
(5) The pharmaceutical composition according to any one of (2) to (4), wherein R ¹ and R ² are independently a C _1-6 alkoxy group, and R ³ is a carboxy group.
(6) The substituted phenanthrene compound is:

The pharmaceutical composition according to (4), which is a compound selected from the group consisting of:
(7) Cancer is pancreatic cancer, stomach cancer, colon cancer, kidney cancer, liver cancer, bone marrow cancer, adrenal cancer, skin cancer, melanoma, lung cancer, small intestine cancer, prostate cancer, testis The pharmaceutical composition according to any one of (1) to (6), which is cancer, uterine cancer, breast cancer or ovarian cancer.
(8) The pharmaceutical composition according to (7), wherein the cancer is pancreatic cancer.

  The present invention provides a compound effective for the prevention and / or treatment of cancer.

It is a graph which shows Pim-1, Pim-2, and Pim-3 inhibitory activity of the substituted phenanthrene compound (T19) of this invention. It is a graph which shows Pim-1, Pim-2, and Pim-3 inhibitory activity of the substituted phenanthrene compound (T19-1) of this invention. It is a graph which shows Pim-1, Pim-2, and Pim-3 inhibitory activity of the substituted phenanthrene compound (T19-2) of this invention. 2 is a graph showing the growth inhibitory activity of the substituted phenanthrene compound (T19) of the present invention on human pancreatic cancer cell lines (PCI55, MiaPaCa-2, PANC-1). 2 is a graph showing the growth inhibitory activity of the substituted phenanthrene compound (T19-1) of the present invention on human pancreatic cancer cell lines (PCI55, MiaPaCa-2, PANC-1). 1 is a graph showing the growth inhibitory activity of a substituted phenanthrene compound (T19-2) of the present invention on human pancreatic cancer cell lines (PCI55, MiaPaCa-2, PANC-1). It is a graph which shows the growth inhibitory activity of the human fetal kidney cell line (HEK293) of the substituted phenanthrene compound (T19-1) of this invention.

In the present invention, phenanthrene is
1 to 9, preferably 2 to 4, more preferably 3 selected from the group consisting of a hydroxy group, a halogen, a C _1-6 alkoxy group, and a C _1-6 alkoxy-C _1-6 alkyl group Substituents (preferably C _1-6 alkoxy groups, more preferably C _1-3 alkoxy groups, especially methoxy groups), carboxy groups, sulfonic acid groups, hydroxy groups, hydroxy C _1-6 alkyl groups, amide groups, ethylene oxide 1 to 3, preferably 1 or 2 hydrophilic groups (preferably carboxy groups) selected from the group consisting of a group, a propylene oxide group, and an ammonium group
The present invention relates to a pharmaceutical composition for preventing and / or treating cancer, comprising, as an active ingredient, a substituted phenanthrene compound substituted with a salt thereof, a solvate, or a physiologically functional derivative thereof. Here, the substituent and the hydrophilic group are preferably present at different positions on the phenanthrene ring. Hereinafter, this substituted phenanthrene compound may be referred to as the substituted phenanthrene compound of the present invention or the compound of the present invention.

で表され、式中、
R¹およびR²は、独立に、ヒドロキシ基、ハロゲン、C_1-6アルコキシ基、およびC_1-6アルコキシ-C_1-6アルキル基からなる群から選択され、より好ましくはC_1-3アルコキシ基、特にメトキシ基であり、
R³は、カルボキシ基、スルホン酸基、ヒドロキシ基、ヒドロキシC_1-6アルキル基、アミド基、エチレンオキシド基、プロピレンオキシド基、およびアンモニウム基からなる群から選択される親水基、好ましくはカルボキシ基であり、
nおよびmは、独立に、0〜4、好ましくは1〜3、より好ましくは1もしくは2であり、ただし、n + mは1〜8、好ましくは2〜5、より好ましくは3もしくは4であり、
pは、1または2、好ましくは1である。 The substituted phenanthrene compounds of the present invention preferably have the formula I:

In the formula,
R ¹ and R ² are independently selected from the group consisting of hydroxy group, halogen, C _1-6 alkoxy group, and C _1-6 alkoxy-C _1-6 alkyl group, more preferably C _1-3 alkoxy A group, in particular a methoxy group,
R ³ is a hydrophilic group selected from the group consisting of a carboxy group, a sulfonic acid group, a hydroxy group, a hydroxy C _1-6 alkyl group, an amide group, an ethylene oxide group, a propylene oxide group, and an ammonium group, preferably a carboxy group. Yes,
n and m are independently 0-4, preferably 1-3, more preferably 1 or 2, provided that n + m is 1-8, preferably 2-5, more preferably 3 or 4. Yes,
p is 1 or 2, preferably 1.

[式中、R¹、R²およびR³は、式Ｉについて記載したとおりであり、
nおよびmは、独立に、0〜4、好ましくは1〜3、より好ましくは1もしくは2であり、ただし、n + mは1〜8、好ましくは2〜5、より好ましくは3もしくは4である]
で表される化合物である。 More preferably, the substituted phenanthrene compound has the formula II:

[Wherein R ¹ , R ² and R ³ are as described for formula I;
n and m are independently 0-4, preferably 1-3, more preferably 1 or 2, provided that n + m is 1-8, preferably 2-5, more preferably 3 or 4. is there]
It is a compound represented by these.

In Formula I and Formula II, when there are a plurality of R ¹ and R ² , respectively, they are preferably present at different positions on the phenanthrene ring.

[式中、R¹、R²およびR³は、式Ｉについて記載したとおりである]
で表される化合物である。 More preferably, the substituted phenanthrene compound has the formula IIa, IIb or IIc:

[Wherein R ¹ , R ² and R ³ are as described for formula I]
It is a compound represented by these.

Specific examples of the substituted phenanthrene compound in the present invention include the following compounds:

  The substituted phenanthrene compounds in the present invention are described in, for example, Pande, H .; Bhakuni, D, SJCS Perkin I 1976, 2197, and Bremmer, ML; Khatri. NA: Weinreb, SMJ Org. Chem. 1983, 48, 3661. It can be synthesized by the method.

  In the present specification, the “alkyl group” refers to a linear or branched saturated hydrocarbon group. Examples of “alkyl” include, but are not limited to, methyl, ethyl, propyl, isopropyl, isobutyl, n-butyl, tert-butyl, isopentyl, n-pentyl.

The number of atoms (such as carbon atoms) is represented, for example, as “C _xy alkyl group”, for example, “C _1-6 alkyl group” refers to an alkyl group having 1 to 6 carbon atoms.

As used herein, “halogen” refers to fluorine, chlorine, bromine, or iodine.
As used herein, the term “alkoxy group” refers to an “—OR ′” group, where R ′ is an alkyl group as described above.

As used herein, the term “C _1-6 alkoxy-C _1-6 alkyl group” is a C _1-6 alkyl group substituted with a C _1-6 alkoxy group, for example, —CH ₂ —O—CH ₃ is mentioned.
As used herein, the term “hydroxy C _1-6 alkyl group” is a C _1-6 alkyl group substituted with a hydroxy group.

As used herein, an “amide group” refers to a substituted or unsubstituted amide group, ie, a “—C (O) NR′R ″” group, wherein R ′ and R ″ are independently H, A C _1-6 alkyl group, a C _2-6 alkenyl group, a C _2-6 alkynyl group, or a C _3-6 cycloalkyl group is represented. Examples of the “amide group” include groups such as —C (O) NH ₂ , —C (O) NH (CH ₃ ) and —C (O) N (CH ₃ ) ₂ .

  In the present specification, the “alkenyl group” refers to a linear or branched aliphatic hydrocarbon group having one or more carbon-carbon double bonds. Examples include, but are not limited to vinyl, allyl, and the like.

  As used herein, “alkynyl group” refers to a linear or branched aliphatic hydrocarbon group having one or more carbon-carbon triple bonds. Examples include, but are not limited to ethynyl and the like.

  As used herein, “cycloalkyl group” refers to a non-aromatic hydrocarbon ring group. “Cycloalkyl groups” include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

As used herein, “ethylene oxide group” refers to a “— (OCH ₂ CH ₂ ) _x H” group, where x is 2 to 6, preferably 3 to 4. In the present specification, the “propylene oxide group” refers to a “— (OCH ₂ CH ₂ CH ₂ ) _y H” group, where y is 2 to 6, preferably 3 to 4.

  The substituted phenanthrene compounds of the present invention may crystallize in one or more forms (known as polymorphisms), and such polymorphic forms are within the scope of the present invention. Polymorphism generally occurs in response to changes in temperature, pressure, or both. Polymorphs can be distinguished by physical properties known in the art, such as X-ray diffraction patterns, solubility, and melting point.

  The substituted phenanthrene compounds of the present invention can have one or more chiral centers or exist as multiple stereoisomers. The scope of the present invention includes mixtures of isomers (eg, stereoisomers, positional isomers, geometric isomers or optical isomers) as well as purified isomers.

  The salt of the substituted phenanthrene compound of the present invention is usually a pharmaceutically acceptable salt. “Pharmaceutically acceptable salt” refers to a non-toxic salt of a compound of the invention. Salts of the compounds of the present invention include acid addition salts. Representative salts include acetate, benzenesulfonate, benzoate, carbonate, sulfate, tartrate, borate, calcium edetate, cansylate, clavulanate, citrate, Edsylate, fumarate, gluconate, glutamate, hydrobromide, hydrochloride, hydroxynaphthoate, isothionate, lactate, lactobionate, laurate, malate, maleic acid Salt, mandelate, mesylate, methyl sulfate, monopotassium maleate, mucinate, napsylate, nitrate, oxalate, palmitate, pantothenate, phosphate / diphosphate , Potassium salt, salicylate, sodium salt, stearate, basic acetate, succinate, tannate, tosylate, ethyl triiodide, trimethylammonium salt, and Valeric acid salt is mentioned.

  As used herein, “solvate” refers to a complex formed by a solute (the substituted phenanthrene compound of the present invention, or a salt thereof or a physiologically functional derivative thereof) and a solvent. Examples of solvents include but are not limited to water, methanol, ethanol, and acetic acid.

  As used herein, “physiologically functional derivative” refers to the present invention capable of providing (directly or indirectly) the substituted phenanthrene compound of the present invention or an active metabolite thereof upon administration to a mammal. And any pharmaceutically acceptable derivative of the substituted phenanthrene compound. Such derivatives, such as esters and amides, are obvious to those skilled in the art without undue experimentation (Burger's Medicinal Chemistry And Drug Discovery, 5th edition, Vol 1: Principles and Practice).

  The substituted phenanthrene compound of the present invention, or a salt, solvate, or physiologically functional derivative thereof is effective in the prevention and / or treatment of cancer. Cancer can be prevented and / or treated by administering an effective amount of a substituted phenanthrene compound of the present invention, or a salt, solvate, or physiologically functional derivative thereof to a patient in need thereof. it can.

  “Effective amount” means the amount of a compound that elicits a biological or medical response of a tissue, system, animal or human, for example as desired by a researcher or clinician. Effective amounts of the substituted phenanthrene compounds of the invention will depend, for example, on the age, weight, severity, nature of the formulation, and route of administration of the patient. An effective amount of a compound of formula I for the treatment of mammals, particularly humans, is usually in the range of 0.1-100 mg / kg / day, for example 0.1-10 mg / kg / day.

  In the present invention, the cancer includes pancreatic cancer, stomach cancer, colon cancer, kidney cancer, liver cancer, bone marrow cancer, adrenal cancer, skin cancer, melanoma, lung cancer, small intestine cancer, prostate cancer. Testicular cancer, uterine cancer, breast cancer or ovarian cancer are included. The pharmaceutical composition for preventing and / or treating cancer of the present invention is particularly effective for preventing and / or treating pancreatic cancer. Since the substituted phenanthrene compound of the present invention has an action of inhibiting Pim-1, Pim-2 and Pim-3, particularly Pim-3 activity, a pharmaceutical composition for preventing and / or treating the cancer of the present invention Are particularly effective in cancers in which Pim-1, Pim-2 and / or Pim-3, in particular Pim-3, is upregulated. The substituted phenanthrene compounds of the present invention can also be used as Pim-1, Pim-2 and Pim-3 activity inhibitors, particularly as Pim-3 activity inhibitors.

  In the present invention, the prevention of a disease includes suppressing and delaying the onset of the disease, and includes not only the prevention before becoming a disease but also the prevention of the recurrence of the disease after treatment. In the present invention, treatment of a disease includes curing the disease, improving symptoms, and suppressing the progression of symptoms.

  The administration target of the pharmaceutical composition of the present invention is preferably a mammal. As used herein, a mammal refers to a warm-blooded vertebrate, for example, primates such as humans and monkeys, rodents such as mice, rats and rabbits, companion animals such as dogs and cats, and cattle, horses and Examples include livestock such as pigs. The pharmaceutical composition of the present invention is suitable for administration to primates, particularly humans.

  The pharmaceutical composition of the present invention comprises a compound represented by Formula I, or a salt, solvate, or physiologically functional derivative thereof, and one or more pharmaceutically acceptable carriers. A pharmaceutically acceptable carrier generally refers to an inert, non-toxic, solid or liquid, bulking agent, diluent or encapsulating material that does not react with the active ingredients of the present invention, for example, Water, ethanol, polyol (eg, glycerol, propylene glycol, liquid polyethylene glycol, etc.), suitable mixtures thereof, solvents or dispersion media such as vegetable oils, and the like.

  The pharmaceutical composition of the present invention can be administered orally, parenterally, e.g., into the skin, subcutaneously, mucous membranes, intravenously, intraarterially, intramuscularly, intraperitoneally, intravaginally, pulmonary, brain. Administered into the eye, into the eye, and intranasally. Examples of oral preparations include tablets, granules, fine granules, powders, capsules, chewables, pellets, syrups, solutions, suspensions and inhalants. As parenteral preparations, suppositories, retention enemas, drops, eye drops, nasal drops, pessaries, injections, mouth washes, ointments, creams, gels, controlled release patches and patches Skin external preparations, and the like. The pharmaceutical compositions of the invention are administered parenterally in the form of sustained-release subcutaneous implants or in the form of targeted delivery systems (eg, monoclonal antibodies, vector delivery, ion implantation, polymer matrices, liposomes and microspheres). May be.

  The pharmaceutical composition of the present invention may further contain additives conventionally used in the pharmaceutical field. Such additives include, for example, excipients, binders, disintegrants, lubricants, antioxidants, colorants, flavoring agents, and the like, and can be used as necessary. In order to achieve sustained release so that it can act for a long time, it can be coated with a known retarder or the like. Examples of excipients include sodium carboxymethylcellulose, agar, light anhydrous silicic acid, gelatin, crystalline cellulose, sorbitol, talc, dextrin, starch, lactose, sucrose, glucose, mannitol, magnesium aluminate metasilicate, calcium hydrogen phosphate Etc. can be used. Examples of the binder include gum arabic, sodium alginate, ethanol, ethyl cellulose, sodium caseinate, sodium carboxymethyl cellulose, agar, purified water, gelatin, starch, tragacanth, lactose, hydroxycellulose, hydroxymethylcellulose, hydroxypropylcellulose, polyvinylpyrrolidone, etc. Is mentioned. Examples of the disintegrant include carboxymethyl cellulose, carboxymethyl cellulose sodium, carboxymethyl cellulose calcium, crystalline cellulose, starch, hydroxypropyl starch and the like. Examples of the lubricant include stearic acid, calcium stearate, magnesium stearate, talc, hydrogenated oil, sucrose fatty acid ester, waxes and the like. Examples of the antioxidant include tocopherol, gallic acid ester, dibutylhydroxytoluene (BHT), butylhydroxyanisole (BHA), ascorbic acid and the like. Other additives and drugs as required, such as antacids (sodium bicarbonate, magnesium carbonate, precipitated calcium carbonate, synthetic hydrotalcite, etc.), gastric mucosa protective agents (synthetic aluminum silicate, sucralfate, copper chlorophyllin sodium, etc.) ) May be added.

Example 1 Synthesis of 2,3,6-trimethoxyphenanthrene-9-carboxylic acid (T19)

Synthesis of (E) -3- (2-bromo-4,5-dimethoxyphenyl) -2- (4-methoxyphenyl) acrylic acid (compound 3)
To a solution of 4-methoxyphenylacetic acid (8.38 g, 50.5 mmol) in tetrahydrofuran (50 ml) was added potassium tert-butoxide (5.76 g, 51.4 mmol), and the mixture was stirred at room temperature for 3 hours to prepare Compound 1. The solvent (tetrahydrofuran) was distilled off, 2-bromo-4,5-dimethoxybenzaldehyde (Compound 2) and acetic anhydride (60 ml) were added, and the mixture was heated at 105 ° C. for 15 hours. Hot water (120 ml) was added, the mixture was stirred at room temperature overnight, and the crystals were collected by filtration. The filtrate was extracted with chloroform, the organic layer was washed with saturated brine and dried, and then the solvent was distilled off. The residue and crystals were combined and recrystallized from ethanol / hexane to give Compound 3 (12.92 g, 70%). Mp 183-185 ° C (Hexane / AcOEt); IR (CHCl ₃ ) ν1685 cm ^-1 ; ¹ H NMR (270 MHz, CDCl ₃ ) δ 3.27 (3H, s), 3.79 (3H, s), 3.85 (3H, s), 6.37 (1H, s), 6.88 (1H, dd, J = 8.7, 2.4 Hz), 6.89 (1H, dd, J = 8.7, 2.5 Hz), 7.01 (1H, s), 7.16 (1H, dd , J = 8.7, 2.4 Hz), 7.17 (1H, dd, J = 8.7, 2.5 Hz), 8.11 (1H, s); ¹³ C NMR (68 MHz, CDCl ₃ ) δ 55.2, 55.3, 56.1, 113.4, 114.1 , 115.0, 117.5, 126.6, 127.1, 131.2, 131.4, 140.6, 147.3, 150.1, 159.3, 173.1. Theoretical values for C ₁₈ H ₁₇ BrO ₅ : C, 54.98; H, 4.36.Observed values: C, 54.58; H, 4.61.

Synthesis of methyl (E) -3- (2-bromo-4,5-dimethoxyphenyl) -2- (4-methoxyphenyl) acrylate (compound 4) Compound 3 (12.22 g, 31.0 mmol) in acetonitrile (95 ml) To the solution, 1,8-diazabicyclo [5,4,0] -7-undecene (7.5 ml, 50.2 mmol) and iodomethane (9.5 ml, 153.0 mmol) were added at room temperature, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with saturated aqueous ammonium chloride and extracted with ethyl ether. The organic layer was washed with a saturated aqueous sodium hydrogen carbonate solution and dried, and then the solvent was distilled off. The residue was recrystallized from hexane / ethyl acetate to obtain Compound 4 (12.33 g, 97%). Mp 106-107.5 ° C (Hexane / AcOEt); IR (CHCl ₃ ) ν 1710 cm ^-1 ; ¹ H NMR (500 MHz, CDCl ₃ ) δ 3.27 (3H, s), 3.78 (3H, s), 3.82 (3H , s), 3.84 (3H, s), 6.32 (1H, s), 6.86 (1H, dd, J = 9.8, 2.4 Hz), 6.87 (1H, dd, J = 9.8, 2.4 Hz), 7.00 (1H, s), 7.12 (1H, dd, J = 9.2, 2.4 Hz), 7.13 (1H, dd, J = 9.2, 2.4 Hz), 7.95 (1H, s); ¹³ C-NMR (68 MHz, CDCl ₃ ) δ52 .9, 55.7, 56.5, 114.0, 114.6, 115.5, 117.4, 127.5, 128.1, 132.0, 132.7, 139.1, 147.8, 150.3, 159.7, 168.7. Theoretical values for C1 ₉ H ₁₉ BrO ₅ : C, 56.03; H, 4.70 Actual value: C, 55.94; H, 4.76.

Synthesis of methyl 2,3,6-trimethoxyphenanthrene-9-carboxylate (compound 5 ) To a refluxing toluene (110 ml) solution of compound 4 (5.96 g, 14.6 mmol), tributyltin hydride (6.0 ml, 22.3 mmol) And 1,1′-azobis (cyclohexane-1-carbonitrile) (2.16 g, 8.84 mmol) in toluene (70 ml) were added dropwise over 1 hour. The mixture was further heated under reflux for 3 hours, the solvent was distilled off, and the residue was purified by silica gel column chromatography containing 10% potassium fluoride to obtain Compound 5 (3.53 g, 74%). Mp 165.5-167.5 ° C (benzene / petroleum ether) [lit.1 Mp 165-166 ° C (benzene / petroleum ether)]; IR (CHCl ₃ ) ν1710 cm ^-1 ; ¹ H NMR (270 MHz, CDCl ₃ ) δ 3.99 (3H, s), 4.00 (3H, s,), 4.01 (3H, s), 4.09 (3H, s), 7.19 (1H, s), 7.25 (1H, dd, J = 9.2, 2.4 Hz), 7.76 (1H, s), 7.80 (1H, d, J = 2.7 Hz), 8.24 (1H, s), 8.92 (1H, d, J = 8.9 Hz); ¹³ C NMR (68 MHz, CDCl ₃ ) δ 52.0, 55.4, 55.9, 55.9, 103.0, 104.2, 109.1, 115.6, 123.2, 123.6, 125.5, 126.6, 128.3, 129.5, 131.6, 149.6, 150.8, 157.9, 168.1.Theoretical values for C ₁₉ H ₁₈ O ₅ : C, 69.93; H, 5.56. Found: C, 69.79; H, 5.61.

Synthesis of 2,3,6-trimethoxyphenanthrene-9-carboxylic acid (Compound 6) Compound 5 (275 mg, 0.675 mmol) was dissolved in methanol (4.4 ml) and tetrahydrofuran (11 ml), and 10% sodium hydroxide Aqueous solution (2.2 ml) was added, and the mixture was stirred at 60 ° C. for 1 hr. After allowing to cool, the reaction mixture was acidified with 10% hydrochloric acid, and extracted with methylene chloride. The organic layer was washed with saturated brine and dried, and the solvent was evaporated. The residue was recrystallized from hexane / ethyl acetate to obtain Compound 6 (252 mg. 96%). ¹ H NMR (500 MHz, CD3OD) δ 3.99 (3H, s), 4.02 (3H, s), 4.09 (3H, s), 7.22 (1H, d, J = 9.3 Hz), 7.40 (1H, s), 7.98 (2H, s), 8.31 (1H, s), 8.86 (1H, d, J = 9.3 Hz)

Example 2 Synthesis of 2,3,6-trimethoxyphenanthrene-10-carboxylic acid (T19-1)

Synthesis of (E) -2- (2-bromo-4,5-dimethoxyphenyl) -3- (4-methoxyphenyl) acrylic acid (compound 9)
p-Anisaldehyde (19.9 ml, 0.164 mol) (7) and 2-bromo-4,5-dimethoxyphenylacetic acid (8) (45.05 g, 0.164 mol) were mixed with acetic anhydride (82 ml) and triethylamine (30 ml). To the mixture was added at room temperature and the mixture was heated at 110 ° C. for 9 hours. Water (100 ml) was added and the mixture was stirred overnight at room temperature. The reaction mixture was extracted with chloroform, the organic layer was dried, and the solvent was distilled off. The residue was dissolved in 10% aqueous sodium hydroxide solution and extracted with ethyl ether. The aqueous layer was acidified with 10% hydrochloric acid and extracted with chloroform. The organic layer was dried and the solvent was distilled off to obtain Compound 9 (37.01 g, 57%). Mp 224.5-225.5 ℃ (AcOEt,); IR (CHCl ₃ ) ν 1680cm ^-1 ; ¹ H NMR (500 MHz, CDCl ₃ ) δ 3.77 (3H, s), 3.78 (3H, s), 3.93 (3H, s ), 6.68 (1H, s), 6.74 (1H, dd, J = 9.8, 2.4 Hz), 6.76 (1H, dd, J = 9.8, 2.4 Hz), 7.04 (1H, dd, J = 9.8, 2.4 Hz) , 7.05 (1H, dd, J = 9.8, 2.4 Hz), 7.15 (1H, s), 7.93 (1H, s); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 55.3, 56.1, 56.2, 113.4, 114.1, 114.6, 115.7, 126.7, 127.8, 128.8, 132.6, 143.1, 149.0, 149.4, 161.1, 172.3.Theoretical values for C ₁₈ H ₁₇ BrO ₅ : C, 54.98; H, 4.36. Actual values: C, 54.76; H, 4.36 .

Synthesis of methyl (E) -2- (2-bromo-4,5-dimethoxyphenyl) -3- (4-methoxyphenyl) acrylate (Compound 10) Compound 9 (37.01 g, 0.094 mol) in acetonitrile (130 ml) To the solution were added 1,8-diazabicyclo [5,4,0] -7-undecene (28.1 ml, 0.189 mol) and iodomethane (39.3 ml, 0.631 mol) at room temperature, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with saturated aqueous ammonium chloride and extracted with ethyl acetate. After the organic layer was dried, the solvent was distilled off. The residue was passed through silica gel column chromatography (hexane / ethyl acetate, 3: 1 → 2: 1), the solvent was distilled off, and recrystallization from hexane / ethyl acetate gave compound 10 (28.37 g, 75%). It was. Mp 115.0-115.5 ° C (Hexane / AcOEt); IR (CHCl ₃ ) ν 1700cm ^-1 ; ¹ H NMR (500 MHz, CDCl ₃ ) δ 3.75 (3H, s), 3.76 (3H, s), 3.79 (3H, s), 3.93 (3H, s), 6.65 (1H, s), 6.72 (1H, dd, J = 10.0, 2.4 Hz), 6.73 (1H, dd, J = 10.0, 2.4 Hz), 7.01 (1H, dd , J = 9.8, 2.4 Hz), 7.02 (1H, dd, J = 9.8, 2.4 Hz), 7.14 (1H, s), 7.83 (1H, s); ¹³ C NMR (125 MHz, CDCl ₃ ) δ52.3 , 55.2, 56.0, 56.1, 113.4, 113.9, 114.5, 115.6, 126.8, 128.7, 129.3, 132.2, 141.1, 148.8, 149.2, 160.6, 167.7. Theoretical value of C ₁₉ H ₁₉ BrO ₅ : C, 56.03; H, 4.70 Actual value: C, 55.83; H, 4.69.

Synthesis of methyl 2,3,6-trimethoxyphenanthrene-10-carboxylate (Compound 11) Compound 10 (8.00 g, 19.65 mmol) and 1,1'-azobis (cyclohexane-1-carbonitrile) (4.80 g, 19.64 mmol) in refluxing chlorobenzene (110 ml), tributyltin hydride (7.9 ml, 29.4 mmol) and 1,1'-azobis (cyclohexane-1-carbonitrile) (4.80 g, 19.64 mmol) in chlorobenzene (70 ml) The solution was added dropwise over 2 hours. The mixture was further heated at reflux for 1 hour, then the solvent was distilled off, and the residue was purified by silica gel column chromatography (hexane / ethyl acetate, 1: 1) containing 10% potassium fluoride to give compound 11 (3.55 g, 56 %)was gotten. Mp 152.5-153.5 ℃ (MeOH) (lit.2 mp 151-152 ℃); ¹ H NMR (500 MHz, CDCl ₃ ) δ 3.99 (3H, s), 4.00 (3H, s), 4.07 (3H, s) , 4.07 (3H, s), 7.16 (1H, dd, J = 8.8, 2.2 Hz), 7.71 (1H, d, J = 2.2 Hz), 7.77 (1H, s), 7.78 (1H, d, J = 8.8 Hz), 8.37 (1H, s), 8.61 (1H, s); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 51.9, 55.4, 55.8, 103.1, 103.5, 106.8, 115.9, 121.5, 124.1, 124.8, 125.0, 131.2, 133.3, 148.7, 149.8, 160.1, 168.1.

Synthesis of 2,3,6-trimethoxyphenanthrene-10-carboxylic acid (compound 12) Compound 11 (76 mg, 0.232 mmol) was dissolved in methanol (1.2 ml) and tetrahydrofuran (3.0 ml), and 10% sodium hydroxide Aqueous solution (0.6 ml) was added, and the mixture was stirred at 60 ° C. for 1 hr. After allowing to cool, the reaction mixture was acidified with 10% hydrochloric acid, and extracted with methylene chloride. The organic layer was washed with saturated brine and dried, and the solvent was evaporated. The residue was recrystallized from hexane / ethyl acetate to obtain Compound 12 (60.0 mg. 83%). ¹ H NMR (270 MHz, (CD3) 2CO) δ 3.97 (3H, s), 4.06 (3H, s), 4.08 (3H, s), 7.28 (1H, d, J = 6.4 Hz), 8.01 (1H, d, J = 8.7 Hz), 8.11 (1H, s), 8.18 (1H, s), 8.57 (1H, s), 8.77 (1H, s)

Example 3 Synthesis of 2,3,8-trimethoxyphenanthrene-10-carboxylic acid (T19-2)

Synthesis of (E) -2- (2-bromo-4,5-dimethoxyphenyl) -3- (2-methoxyphenyl) acrylic acid (Compound 14)
o-Anisaldehyde (0.725 ml, 6.00 mmol) (Compound 13) and 2-bromo-4,5-dimethoxyphenylacetic acid (Compound 8) (1.65 g, 6.00 mmol) were mixed with acetic anhydride (6.0 m) and triethylamine (3.0 ml). ) At room temperature and the mixture was heated at 110 ° C. for 4 hours. Water (20 ml) was added and the mixture was stirred at 50 ° C. for 1 hour. The reaction mixture was extracted with methylene chloride, the organic layer was dried and the solvent was distilled off. The residue was dissolved in 10% aqueous sodium hydroxide solution and extracted with ethyl ether. The aqueous layer was acidified with 10% hydrochloric acid and extracted with methylene chloride. The organic layer was dried and the solvent was distilled off. The residue was recrystallized from hexane / ethyl acetate to obtain Compound 14 (997 mg, 42%).

Synthesis of methyl (E) -2- (2-bromo-4,5-dimethoxyphenyl) -3- (2-methoxyphenyl) acrylate (compound 15) Compound 14 (997 mg, 2.54 mmol) in acetonitrile (7.0 ml) To the solution, 1,8-diazabicyclo [5,4,0] -7-undecene (0.570 ml, 3.81 mmol) and iodomethane (0.791 ml, 12.7 mmol) were added at room temperature, and the mixture was stirred at room temperature for 1 hour. Saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl ether. After the organic layer was dried, the solvent was distilled off. The residue was passed through silica gel column chromatography (hexane / ethyl acetate, 3: 1), the solvent was distilled off, and recrystallization from hexane / ethyl acetate gave compound 15 (737 mg, 72%). ¹ H NMR (500 MHz, CDCl ₃ ) δ 3.67 (3H, s), 3.81 (3H, s), 3.87 (3H, s), 3.90 (3H, s), 6.57 (1H, s), 6.64 (1H, t, J = 7.6 Hz), 6.72 (1H, d, J = 6.3 Hz), 6.85 (1H, d, J = 8.3 Hz), 7.10 (1H, s), 7.21 (1H, t, J = 7.6 Hz) , 8.22 (1H, s)

Synthesis of methyl 2,3,8-trimethoxyphenanthrene-10-carboxylate (Compound 16) Compound 15 (737 mg, 1.81 mmol) and 1,1'-azobis (cyclohexane-1-carbonitrile) (442 mg, 1.81 mmol) in refluxing chlorobenzene (10 ml), tributyltin hydride (0.732 ml, 2.72 mmol) and 1,1'-azobis (cyclohexane-1-carbonitrile) (442 mg, 1.81 mmol) in chlorobenzene (5.0 ml) The solution was added dropwise over 3 hours. The solvent was distilled off, and the residue was passed through silica gel column chromatography (hexane / ethyl acetate) containing 10% potassium fluoride. Recrystallization from hexane / ethyl acetate gave compound 16 (230 mg, 39%). . ¹ H NMR (270 MHz, CDCl ₃ ) δ 4.03 (3H, s), 4.06 (3H, s), 4.09 (3H, s), 4.12 (3H, s), 6.94 (1H, d, J = 7.9 Hz) , 7.62 (1H, t, J = 8.2 Hz), 7.98 (1H, s), 8.09 (1H, d, J = 8.4 Hz), 8.62 (1H, s)

Synthesis of 2,3,8-trimethoxyphenanthrene-10-carboxylic acid (compound 17) Compound 16 (180 mg, 0.552 mmol) was dissolved in methanol (2.8 ml) and tetrahydrofuran (7.0 ml), and 10% sodium hydroxide Aqueous solution (1.4 ml) was added, and the mixture was stirred at 60 ° C. for 1 hr. After allowing to cool, the reaction mixture was acidified with 10% hydrochloric acid, and extracted with methylene chloride. The organic layer was washed with saturated brine and dried, and the solvent was evaporated. The residue was recrystallized from hexane / ethyl acetate to give 17 (130 mg, 76%). ¹ H NMR (270 MHz, (CD ₃ ) ₂ CO) δ 3.96 (3H, s), 4.09 (6H, s), 7.12 (1H, d, J = 7.6 Hz), 7.68 (1H, t, J = 8.2 Hz), 8.21 (1H, s), 8.32 (1H, d, J = 8.1 Hz), 8.72 (1H, s), 9.05 (1H, s)

(Example 4) In vitro assay
Carbonate buffer (pH 9.6) [15 μmol / L Na ₂ CO ₃ (Wako), 35 μmol / L NaHCO ₃ (Wako) on a 96-well microplate (Nunc-Immuno ^™ Plate, NUNC Brand Products, Nalge Nunc International, Denmark) His-Bad protein solution (3 μg / mL) diluted with 0.02% (w / v) NaN ₃ (Wako) pH 9.6] was added at 100 μL / well and coated overnight at 4 ° C. On the next day, the plate was washed 3 times, 1% BSA / PBS (−) solution was added at 150 μL / well, blocked at 37 ° C. for 1 hour, and washed 3 times. 19.2 nmol / L thioredoxin-hexahistidine-Pim (Pim-1, Pim-2 or Pim-3) fusion protein, 10 μmol / L ATP (Cell Signaling), compound T19 synthesized in the above example, Kinase buffers [25 mmol / L Tris-HCl (pH 7.5), 10 mmol / L MgCl ₂ , 0.1 mmol / L Na ₃ VO ₄ , 2 mmol / L dithiothrease containing various concentrations of T19-1 and T19-2 respectively] Tall (DTT), 5 mmol / L s-glycerophosphate] (Cell Signaling) was added at 40 μL / well. After reacting at 30 ° C. for 1 hour, washing was performed 5 times, 1,000-fold diluted anti-pBadser112 rabbit antibody (Cell Signaling) was added at 100 μL / well, and reacted at 37 ° C. for 1 hour. After washing 5 times, an alkaline phosphatase-labeled anti-rabbit immunoglobulin antibody diluted 10,000 times was added at 100 μL / well, and further reacted at 37 ° C. for 1 hour. After washing 10 times, substrate solution (pH 9.8) [0.1% p-nitrophenyl phosphate (Wako), 1 mol / L diethanolamine (Wako)] was added at 100 μL / well and reacted at room temperature for 20-30 minutes. Stop solution [3N NaOH (Wako)] was added at 100 μL / well to terminate the reaction. The absorbance of the plate at a wavelength of 405 nm was measured with a microplate reader (Model 550, Bio-Rad). In addition, 0.05% Tween / PBS (-) was used as a washing buffer and washed with a microplate washer (Model 1575 ImmunoWash, Bio-Rad).
The results are shown in FIGS. Compounds T19, T19-1 and T19-2 were all shown to suppress the activities of Pim-1, Pim-2 and Pim-3 in a concentration-dependent manner.

(Example 5) Cell proliferation assay (pancreatic cancer cells)
A human pancreatic cancer cell line (PCI55, MiaPaCa-2, PANC-1) having 3,000 cells in each well was dispensed into a 96-well cell culture microtest plate (Becton Dickinson) in a medium amount of 100 μL. After culturing for 16 to 18 hours, the medium is removed, and the medium containing the test compound [0.1% DMSO, T19 (50, 75, 100, 125 μM), T19-1 (25, 50, 75, 100 μM), T19-2 (50 , 75, 100, 125 μM)] was added at 100 μL / well. The culture was continued for 0, 24, 48, 72, and 96 hours. After the passage, the test compound-containing medium was removed, and 10% WST-1 (Roche) medium was added at 100 μL / well. After further incubation for 1 hour, absorbance at a wavelength of 450 nm was measured with a microplate reader (Model 550, Bio-Rad). As the medium, RPMI-1640 medium (SIGMA) was used, and 10% calf serum (Invitrogen), 50 U / mL penicillin G (SIGMA), and 50 μg / mL streptomycin (SIGMA) were added. The cell line was cultured in a carbon dioxide cell culture apparatus (ESPEC) at 37 ° C. and a carbon dioxide concentration of 5%.
The results are shown in FIGS. Compounds T19, T19-1 and T19-2 were shown to inhibit the growth of all tested human pancreatic cancer cell lines (PCI55, MiaPaCa-2, PANC-1).

(Example 6) Cell proliferation assay (kidney cancer cells)
A human fetal kidney cell line (HEK293, number of cells in each well 4000) was dispensed in a 96-well cell culture microtest plate (Becton Dickinson) at a medium volume of 100 μL. After culturing for 16 to 18 hours, the medium was removed, and a medium containing the test compound [0.1% DMSO, T19-1 (25, 50, 75, 100 μM)] was added at 100 μL / well. The culture was continued for 0, 24, 48, 72, and 96 hours. After the passage, the test compound-containing medium was removed, and 10% WST-1 (Roche) medium was added at 100 μL / well. After further incubation for 1 hour, absorbance at a wavelength of 450 nm was measured with a microplate reader (Model 550, Bio-Rad). RPMI-1640 medium (SIGMA) was used as the medium, and 10% calf serum (Invitrogen), 50 U / mL penicillin G (SIGMA), and 50 μg / mL streptomycin (SIGMA) were added to each medium. used. The cell line was cultured in a carbon dioxide cell culture apparatus (ESPEC) at 37 ° C. and a carbon dioxide concentration of 5%.
The results are shown in FIG. T19-1 was shown to suppress the growth of the human fetal kidney cell line (HEK293).

Claims (8)

The phenanthrene has 1 to 9 substituents selected from the group consisting of a hydroxy group, a halogen, a C _1-6 alkoxy group, and a C _1-6 alkoxy-C _1-6 alkyl group, and a carboxy group and a sulfonic acid group. A substituted phenanthrene compound substituted with 1 to 3 hydrophilic groups selected from the group consisting of hydroxy group, hydroxy C _1-6 alkyl group, amide group, ethylene oxide group, propylene oxide group, and ammonium group, or A pharmaceutical composition for preventing and / or treating cancer comprising a salt, solvate, or physiologically functional derivative as an active ingredient. Substituted phenanthrene compounds have the formula I:

[Where
R ¹ and R ² are independently a substituent selected from the group consisting of a hydroxy group, a halogen, a C _1-6 alkoxy group, and a C _1-6 alkoxy-C _1-6 alkyl group;
R ³ is a hydrophilic group selected from the group consisting of a carboxy group, a sulfonic acid group, a hydroxy group, a hydroxy C _1-6 alkyl group, an amide group, an ethylene oxide group, a propylene oxide group, and an ammonium group;
n and m are independently 0-4, provided that n + m is 1-8,
p is 1 or 2]
The pharmaceutical composition of Claim 1 which is a compound represented by these. The substituted phenanthrene compound has the formula II:

[Where
R ¹ , R ² and R ³ are as defined in claim 2;
n and m are independently 1 or 2, where n + m is 3]
The pharmaceutical composition of Claim 2 which is a compound represented by these. The substituted phenanthrene compound is of formula IIa, IIb or IIc:

[Wherein R ¹ , R ² and R ³ are as defined in claim 2]
The pharmaceutical composition of Claim 3 which is a compound represented by these. The pharmaceutical composition according to any one of claims 2 to 4, wherein R ¹ and R ² are independently a C _1-6 alkoxy group and R ³ is a carboxy group. Substituted phenanthrene compounds are:

The pharmaceutical composition according to claim 4, which is a compound selected from the group consisting of:   Cancer is pancreatic cancer, stomach cancer, colon cancer, kidney cancer, liver cancer, bone marrow cancer, adrenal cancer, skin cancer, melanoma, lung cancer, small intestine cancer, prostate cancer, testicular cancer, The pharmaceutical composition according to any one of claims 1 to 6, which is uterine cancer, breast cancer or ovarian cancer.   The pharmaceutical composition according to claim 7, wherein the cancer is pancreatic cancer.

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