KR100666207B1 - 6-Alkylamino-2,2'-Disubstituted-Benzopyrane Derivatives as RTK Inhibitor - Google Patents

Abstract

The present invention provides a 6-alkylamino-2,2'-disubstituted-benzopyran derivative represented by the following Chemical Formula 1 having excellent RTK (Receptor Tyrosine Kinase) inhibitory activity, an isomer thereof, and a pharmaceutically acceptable thereof. It relates to the use of the salt as an anticancer agent.

In Formula 1, R ¹ , R ² , R ³ , R ⁴ , n are as defined in the detailed description of the invention, respectively.

Aminobenzopyran, receptor tyrosine enzyme, epithelial cell, anticancer agent, RTK

Description

6-Alkylamino-2,2'-Disubstituted-Benzopyrane Derivatives as RTK Inhibitor}

The present invention provides a 6-alkylamino-2,2'-disubstituted-benzopyran derivative represented by the following Chemical Formula 1 having excellent RTK (Receptor Tyrosine Kinase) inhibitory activity, an isomer thereof, and a pharmaceutically acceptable thereof. It relates to the use of the salt as an anticancer agent.

[Formula 1]

In Chemical Formula 1,

R ¹ is a C ₁ -C ₁₀ alkyl group, benzyl or substituted benzyl group, phenethyl group, 2-pyrimidylmethyl group, thiophene group, 2-methylthiophenmethyl group, 5-methyl-2-thiophenmethyl group, 3- Thiophenmethyl group, indolylmethyl group, benzodioxoranylmethyl group, naphthalenylmethyl group, or furanylmethyl group; R ² and R ³ each represent a hydrogen atom, an alkyl group of C _{1 to} C ₅ , a halogen, or a phenyl and substituted phenyl group; R ⁴ represents a 5 to 7 membered hetero ring containing a hetero atom selected from a C ₁ to C ₁₀ alkyl group, a phenyl or substituted phenyl group, or an oxygen or sulfur atom; And groups wherein a phenyl group or a heterocyclic substituent selected from an alkoxy group of C ₁ ~C ₆ alkyl group, C ₁ ~C ₆ haloalkyl group, a halogen, a nitro group, a cyano group and a C ₁ ~C ₆ of the 1-4-substituted N may represent an integer of 1 to 5.

The present invention has been filed with the invention of Korean Patent Application No. 2004-16206 (US Patent Application 10/970046, Japanese Patent Application 2004/357870), 6-alkylamino-2,2'- It is a novel use invention of a bisubstituted-benzopyran derivative.

Korean Patent Application No. 2004-16206 is related to the 6-alkylamino-2,2'-disubstituted-benzopyran derivative represented by Chemical Formula 1 because it shows an excellent 5-lipoxygenase (5-LO) inhibitory effect. It is also a patent pending invention for use as a prophylactic and therapeutic agent for inflammatory diseases, rheumatoid arthritis, colitis, asthma, psoriasis and the like caused by the activity of leukotriene (LTA4, B4, C4, D4).

The inventors of the present invention to determine the RTK (Receptor Tyrosine Kinase: Receptor Tyrosine Enzyme) inhibitory activity on the 6-alkylamino-2,2'-disubstituted-benzopyran derivative represented by Formula 1, EGFR (epidermal) growth factor receptor (EGP), vascular endotherial growth factor receptor 2 (VEGFR2), platelet-derived growth factor receptor b (PDGFRb), and platelet-derived growth factor receptor (FGFR1), and fibroblast growth Experiments were conducted to confirm the inhibitory activity of tactor receptor 1 (fibroblast growth factor receptor). As a result, the 6-alkylamino-2,2'-disubstituted-benzopyran derivative represented by the formula (1) has been applied for patenting the present invention by confirming that it has excellent inhibitory activity against the RTK-related receptor.

Current anticancer drugs are not complete in terms of efficacy and toxicity. For that reason, it is necessary to overcome the problems of existing anticancer drugs and to develop anticancer drugs having excellent activity.

In connection with the development of new anti-cancer agents for RTK inhibition that can overcome the problems with existing anticancer agents, various heterocyclic compounds have been known. As an example, the present inventors have applied for patents for novel quinoxaline-based heterocyclic compounds having RTK inhibitory activity in Korean Patent Application Nos. 2004-94232 and 2004-94233.

However, since the benzopyran-based heterocyclic compound characterized by the present invention has never been utilized in the development of anticancer agents through the inhibition of RTK activity, the present invention is a 6-alkylamino-2,2'-disubstituted-benzopyran derivative Patent application for a new use of the drug as an anticancer agent.

SUMMARY OF THE INVENTION Since the 6-alkylamino-2,2'-disubstituted-benzopyran derivatives exhibit excellent RTK inhibitory activity, an object of the present invention is to provide a use of the compound as an anticancer agent.

The present invention is characterized by an anticancer agent containing 6-alkylamino-2,2'-disubstituted-benzopyran derivatives represented by the following formula (1), an isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

In Chemical Formula 1,

R ¹ is a C ₁ -C ₁₀ alkyl group, benzyl or substituted benzyl group, phenethyl group, 2-pyrimidylmethyl group, thiophene group, 2-methylthiophenmethyl group, 5-methyl-2-thiophenmethyl group, 3- Thiophenmethyl group, indolylmethyl group, benzodioxoranylmethyl group, naphthalenylmethyl group, or furanylmethyl group; R ² and R ³ each represent a hydrogen atom, an alkyl group of C _{1 to} C ₅ , a halogen, or a phenyl and substituted phenyl group; R ⁴ represents a 5 to 7 membered hetero ring containing a hetero atom selected from a C ₁ to C ₁₀ alkyl group, a phenyl or substituted phenyl group, or an oxygen or sulfur atom; And groups wherein a phenyl group or a heterocyclic substituent selected from an alkoxy group of C ₁ ~C ₆ alkyl group, C ₁ ~C ₆ haloalkyl group, a halogen, a nitro group, a cyano group and a C ₁ ~C ₆ of the 1-4-substituted N may represent an integer of 1 to 5.

The 6-alkylamino-2,2'-disubstituted-benzopyran derivative represented by Chemical Formula 1 according to the present invention has chiral carbon, and thus the present invention is separated by a racemic compound or a conventional separation method. Each isomeric compound and all of these isomeric mixtures are included.

Preparation of 6-alkylamino-2,2'-disubstituted-benzopyran derivatives represented by Formula 1 contained as an active ingredient of an anticancer agent according to the present invention, Korean Patent Application No. 2004-16206 (US Patent Application No. 10 / 970046, Japanese Patent Application 2004/357870) can be synthesized by the same process as the combination chemical synthesis technique described.

In Scheme 1, R ¹ , R ² , R ³ , R ⁴ and n are each as defined above; Ⓟ represents a solid support in the form of a high molecular polymer selected from polystyrene-divinylbenzene, methacrylic acid-dimethylacrylamide and hydroxyl methacrylic acid.

As an active substance constituting the anticancer agent according to the present invention, a pharmaceutically acceptable salt of the 6-alkylamino-2,2'-disubstituted-benzopyran derivative represented by Chemical Formula 1 may be used in a conventional method in the art. It may be prepared by, for example, salts with inorganic acids such as hydrochloric acid, hydrogen bromide, sulfuric acid, sodium hydrogen sulfate, phosphoric acid, carbonic acid or formic acid, acetic acid, oxalic acid, benzoic acid, citric acid, tartaric acid, gluconic acid, gesty acid Together with organic acids such as fumaric acid, lactobionic acid, salicylic acid, or acetylsalicylic acid (aspirin), to form salts of pharmaceutically acceptable acids thereof, or to react with alkali metal ions such as sodium or potassium These metal salts may be formed or reacted with ammonium ions to form another form of a pharmaceutically acceptable salt.

In addition, the anticancer agent of the present invention is a conventional non-toxic pharmaceutically acceptable carrier, adjuvant and excipient in addition to the active substance, such as tablets, capsules, troches, solutions, suspensions, etc. It may be formulated as a preparation for oral administration or a preparation for parenteral administration. In addition, the dosage of the active substance according to the present invention to the human body may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, and is generally based on an adult patient having a weight of 70 kg. In the range of 0.01 to 1000 mg / day, and may be administered once or several times a day at regular intervals according to the judgment of a doctor or pharmacist.

As described above, the present invention will confirm the anticancer activity of the 6-alkylamino-2,2'-disubstituted-benzopyran derivative represented by Formula 1 according to the following examples, but is represented by Formula 1 Specific examples of the preparation method of the compound and the preparation method thereof are intended to be replaced by Korean Patent Application No. 2004-16206 (US Patent Application 10/970046, Japanese Patent Application 2004/357870).

Example. Inhibition of RTK (receptor tyrosine kinase)

Epidermal growth factor receptor (EGFR), vascular endotherial growth factor receptor 2 (VEGFR2), platelet-derived growth factor receptor b (PDGFRb), platelet-derived growth factor receptor (FGFR1) Tyrosine kinase activity of fibroblast growth tactor receptor 1 (fibroblast growth factor receptor) was measured by Alpha Screen ^™ P-Tyr-100 assay method (Perkin Elmer).

AlphaScreen refers to Amplified Luminescent Proximity Homogeneous Assay and is an Immunoassay method using Luminescent Oxygen Radical Channel. In this experiment, biotinylated poly [Glu: Tyr] (4: 1) (Packard BioScience) was used as a substrate for the enzyme, and a streptavidin-coated donor could bind to these substrates. Receptor beads bound to antibodies (P-Tyr-100) capable of recognizing and reacting beads and phosphorylated substrates were used. EGFR was purchased from Sigma (St. Louse, MO, USA), and VEGFR2, PDGFRb, and FGFR1 were purchased from ProQuinase (Germany), respectively.

The assay (Assay) was carried out through the following three steps using a white 384 well plate (Greiner Bio-One). First, RTK enzyme (EGFR 5 ng / VEGFR 2.5 ng / PDGFR 10 ng / FGFR 2.5 ng) was added to each well of a 384 well plate with Tyrphostin A51 (Upstate Biotech), a known standard inhibitor, or the compound to be tested. The reaction was carried out at room temperature for 15 minutes. Second, after 15 minutes of reaction, 5 μL of biotinylated poly [Glu: Tyr] (4: 1) and ATP were added to the final concentrations of 5 nM and 100 mM, respectively, for 1 hour (PDGFR). 2 hours) was subjected to the enzyme reaction at room temperature. The composition of the buffer used for the enzyme reaction was 50 mM Tris-HCl (pH7.5), 5 mM MgCl ₂ , 5 mM MnCl ₂ , 2 mM DTT and 0.01% Tween-20. Third, after the enzymatic reaction, 10 μL of capture buffer containing donor and acceptor beads (20 mg / ml each) was added thereto and left at room temperature for 1 hour. This process takes place in the dark and stops the ongoing enzymatic reaction and recognizes the phosphorylated substrate of the donor-streptavidin and the receptor P-Tyr-100 beads. The composition of the capture buffer (2.5-fold concentration) is 62.5 mM HEPES (pH 7.4), 250 mM NaCl, 100 mM EDTA and BSA 0.25%.

After the reaction was completed, the activity of the enzyme was determined by measuring the AlphaScreen signal using a Fusion ^TM microplate analyzer (PerkinElmer). The experiment to see the dose-reseponse was performed in triple. In other words, the inhibitor was diluted to determine the% inhibition rate at 6 or more concentrations, and the obtained result was obtained by nonlinear regression fitting using the program Prism ^TM (GraphPad software, San Diego, Calif.) To obtain the IC ₅₀ value.

The structures of representative compounds applied in this experiment are shown in Table 1 below. In addition, the drug test results for each test compound are shown in Table 2 below.

As described above, the novel compound represented by Chemical Formula 1 according to the present invention has excellent inhibitory activity against RTK and is useful for development as an anticancer agent.

Claims (1)

An anticancer agent characterized in that it contains a 6-alkylamino-2,2'-disubstituted-benzopyran derivative represented by the following formula (1), or a pharmaceutically acceptable salt thereof: [Formula 1]

In Chemical Formula 1, R ¹ is a C ₁ -C ₁₀ alkyl group, benzyl or substituted benzyl group, phenethyl group, 2-pyrimidylmethyl group, thiophene group, 2-methylthiophenmethyl group, 5-methyl-2-thiophenmethyl group, 3- Thiophenmethyl group, indolylmethyl group, benzodioxoranylmethyl group, naphthalenylmethyl group, or furanylmethyl group; R ² and R ³ each represent a hydrogen atom, an alkyl group of C _{1 to} C ₅ , a halogen, or a phenyl and substituted phenyl group; R ⁴ represents a C ₁ to C ₁₀ alkyl group, a phenyl group or a substituted phenyl group; And the substituted phenyl group or substituted benzyl group is a substituent selected from C _{1 to} C ₆ alkyl group, C _{1 to} C ₆ haloalkyl group, halogen, nitro group, cyano group and C _{1 to} C ₆ alkoxy group. 4 substituted phenyl group or benzyl group, n represents the integer of 1-5.