WO2007066615A1

WO2007066615A1 - Novel activator of nuclear orphan receptor and use thereof

Info

Publication number: WO2007066615A1
Application number: PCT/JP2006/324171
Authority: WO
Inventors: Toshiyuki Shimizu; Takuro Niwa; Kan Chiba; Masakiyo Hosokawa; Kaoru Kobayashi
Original assignee: Mitsubishi Tanabe Pharma Corporation; National University Corporation Chiba University
Priority date: 2005-12-05
Filing date: 2006-12-04
Publication date: 2007-06-14
Also published as: JP2009120486A

Abstract

Disclosed is a compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof or a solvate of the compound or the salt, which can be used as a pregnane receptor activator. (I) wherein R1 represents a cyclohexyl group, a phenyl group which may have at least one substituent selected from a C1-C4 alkyl group, a C1-C4 alkoxy group and a halogen atom, a thienyl group which may have at least one substituent selected from a C1-C4 alkyl group, a C1-C4 alkoxy group and a halogen atom, a furyl group which may have at least one substituent selected from a C1-C4 alkyl group, a C1-C4 alkoxy group and a halogen atom, a thiazolyl group, a phenoxy group, a C1-C4 phenylalkyl group, a phenylthio group, a morpholino group, a piperidyl group, a pyrrolidinyl group, a pyridyl group, or an imidazolyl group; R2 represents -CHR3R4 (where R3 represents a hydrogen atom or a C1-C4 alkyl group; and R4 represents a C1-C4 alkyl group, a cyclohexyl group, a thienyl group or a phenyl group which may have at least one substituent selected from a C1-C4 alkyl group, a C1-C4 alkoxy group and a halogen atom) or a cyclohexyl group; and the ring A represents a benzene ring, a thiphene ring or a furan ring.

Description

Novel activators of nuclear orphan receptors and their uses

Technical field

[0001] The present invention relates to a novel activator of nuclear orphan receptors and uses thereof. More specifically, the present invention relates to a predanan

Background technology

[0002] PXR is one of the nuclear receptors that binds to the transcriptional regulatory sequence of a target gene and activates transcription of the target gene when a ligand binds to it. Because its in vivo ligand has not been clarified, it is classified as an orphan receptor.Rifampicin and lithocholic acid are known to increase the expression of target genes such as CYP (cytochrome P450) 3A4 through PXR. , Ru (Non-Patent Document 1).

CYP3A4 is constantly expressed in the liver and small intestine, and is known to be induced by lithocholic acid, a metabolite of cholesterol, and rifampicin, an anti-tuberculous drug. Also

, CYP3A4 has the effect of acidifying and excreting not only various xenobiotics but also bile acids, lithocholic acid, and dioxycholic acid in the body. There are reports that administering rifampicin, an inducer of CYP3A4, to patients with cholestatic liver disease reduces the amount of excess bihirbin in the body, resulting in a therapeutic effect. There are reports that it is also effective in treating severe itching observed in cases of liver cirrhosis (Non-Patent Document 2). Based on these facts, it is thought that if target genes such as CYP3A4 can be activated through PXR, it will be useful as a therapeutic agent for these diseases.

On the other hand, although various pharmacological effects have been reported for phthalazine derivatives (Patent Documents 1 to 4), it was not known that phthalazine derivatives activate PXR or induce CYP3A4. .

Patent document 1: Japanese Patent Application Publication No. 60-243074

Patent document 2: Japanese Patent Application Laid-Open No. 6-135938 (EP0534443B1)

Patent document 3: Japanese Patent Application Laid-Open No. 8-34734 (EP0682947B1) Patent Document 4: Japanese Unexamined Patent Publication No. 10-109936

Non-patent document 1: Gastroenterology. 2005 Aug;129(2):735- 40.

Non-patent document 2 Gastroenterology. 2005 Aug;129(2):476- 85.

Disclosure of invention

[0003] An object of the present invention is to provide a PXR activator useful as a therapeutic agent for liver diseases and metabolic diseases.

[0004] The inventors of the present invention have conducted extensive studies to solve the above problems. As a result, the following general formula (I

The present inventors have discovered that the compound shown in ) activates PXR, and have completed the present invention.

[0005] That is, the present invention is as follows.

(1) A Predanan X receptor activator containing a compound represented by the following general formula (I), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[C1]

In the above formula, R ¹ is a cyclohexyl group; C to C alkyl group, C to C alkoxy

1 4 1 4

A phenol group, optionally with one or more substituents selected from C to C alkyl groups, C to C alkoxy groups and halogen atoms.

1 4 1 4

Cher group which may have the above substituents; alkyl group of c to c, alkyl group of c to c

1 4 1 4 Lukoxy group and nourogen nuclear power Furyl group which may have one or more selected substituents; thiazolyl group; phenoxy group;

7 9

Represents a rufolino group; a piperidyl group; a pyrrolidyl group; a pyridyl group; or an imidazolyl group,

R ² is CHR ³ R ⁴ (R ³ represents a hydrogen atom or a C to C alkyl group, R ⁴ is a C to C

1 4 1 alkyl group; cyclohexyl group; chel group; or C ~ C alkyl group, C ~

4 1 4 1

Alkoxy group and nozzle group of C may have one or more selected substituents.

Four

represents a phenyl group. ); or represents a cyclohexyl group, Ring A represents a benzene ring, a thiophene ring or a furan ring.

(2) Predanane X receptor activity of (1) containing 1-(1-cyclohexylethylamino)-4-phelphthalazine or a pharmaceutically acceptable salt thereof, or a solvate thereof agent.

(3) Predanane X receptor activator of (1) containing (R) -l-(l-cyclohexylethylamino)-4-phelphthalazine fumarate.

(4) A CYP3A4 inducer containing any of the predanane X receptor activators of (1) to (3).

(5) A liver disease therapeutic agent containing any of the predanan X receptor activators of (1) to (3).

(6) The liver disease treatment drug according to (5), wherein the liver disease is cholestatic liver disease, jaundice, or liver failure.

(7) A liver fibrosis inhibitor containing any of the predanan X receptor activators listed in (1) to (3).

(8) A therapeutic agent for acute kidney disease containing any of the predanan X receptor activators listed in (1) to (3).

(9) A therapeutic agent for metabolic diseases containing any one of the predanan X receptor activators of (1) to (3). Provided is a method of activating Predanane X receptor, which comprises administering an acceptable salt or solvate thereof.

The present invention also provides the use of a compound represented by the above general formula (I), a pharmaceutically acceptable salt thereof, or a solvate thereof for the production of a Predanane X receptor activator. .

Brief description of the drawing

[0006] [Figure 1] Diagram showing the amount of CYP3A4 mRNA when rifampicin (RIF) or Compound 1 was added to cells.

[Figure 2] Diagram showing concentration-dependent activation of a reporter gene by rifampicin (RIF) or Compound 1.

BEST MODE FOR CARRYING OUT THE INVENTION

[0007] The PXR activator of the present invention contains a compound of general formula (I), a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient.

[Case 2]

Here, R ¹ is a cyclohexyl group; a C to C alkyl group, a C to C alkoxy group, and

1 4 1 4

a fer group having one or more substituents selected from halogen atoms and halogen atoms;

1 4 alkyl groups, C to C alkoxy groups and halogen atoms One or more selected

14

Cher group which may have a substituent; c to c alkyl group, c to c alkoxy group

1 4 1 4

and norogen atomic force optionally having one or more substituents selected, furyl group; thiazolyl group; phenoxy group; C to C phenol alkyl group; phenolthio group;

7 9

Represents a piperidyl group; a pyrrolidyl group; a pyridyl group; or an imidazolyl group,

1 4 1 4 alkyl group; cyclohexyl group; chel group; or c to c alkyl group,

1 4 c~

1 c Alkoxy group and nozzle may have one or more selected substituents

Four

represents a strong fer group. ); or represents a cyclohexyl group,

Ring A represents a benzene ring, a thiophene ring or a furan ring.

In the above general formula, C to C alkyl groups include methyl group, ethyl group, n propyl group.

14

group, i-propyl group, n-butyl group, t-butyl group, etc., and C to C alkoxy

14

Examples of the group include methoxy group, ethoxy group, n-propoxy group, i-propoxy group, n-butoxy group, t-butoxy group, etc., and examples of the halogen atom include fluorine atom, chlorine atom, bromine atom, etc.

As R ¹ , a phenol group, a 2-cher group or a 2-furyl group is preferred, and a phenyl group is particularly preferred. As R ² — CHR ³ ' R ⁴ (R ³ represents a C to C alkyl group, R ⁴

14

' represents a cyclohexyl group. ) is especially preferred

[C3] One CH (CH ₃ )

is preferred. As ring A, a compound representing a benzene ring or a thiophene ring is preferred, and a benzene ring is particularly preferred.

[0009] The compound represented by the above general formula (I) is a known compound, and its manufacturing method is not particularly limited. It can be produced according to the method described in JP-A No. 10-109936 and the like.

[0010] As the pharmaceutically acceptable salt of the aminoviridazine derivative of general formula (I), an acid addition salt or a base addition salt can be used. There are no particular limitations.

Since the compound of general formula (I) and its salts may exist in the form of solvates (including hydrates), these solvates (including hydrates) can also be used for the PXR activation of the present invention. It can be used as an active ingredient in drugs. A particularly preferred salt is fumaric acid.

Furthermore, when an asymmetric carbon exists in the aminopyridazine derivative of the above general formula (I), it can take any of the (R) form, (S) form, and (RS) form, all of which are included in the PXR of the present invention. It can be used as an active ingredient in active ingredients.

[0011] Preferred examples of compounds of general formula (I) that can be used as active ingredients of PXR activators include, specifically, compounds represented by the following formula: 1-(1-cyclohexyl tylamino) -4-phenylphthalazine. This compound will be referred to as Compound 1 in the Examples below. More preferred is the R form of Compound 1, and the most preferred example is (R)-l_(l-cyclohexylethylamino)-4-phelphthalazine fumarate.

[C4]

In addition, compounds such as those exemplified in Examples can also be used as active ingredients of PXR activators.

[0012] A compound of general formula (I) or a pharmaceutically acceptable salt thereof or a solvate thereof, either as is or in combination with a pharmaceutically and pharmaceutically acceptable carrier, can be used as a CYP3A4 inducer. Furthermore, it can be used as a therapeutic agent for diseases that can be treated through induction of CYP3A4 and induction of Z or other PXR target genes.

Diseases that can be treated using the PXR activator of the present invention include stasis liver disease, jaundice, liver diseases such as liver fibrosis and liver failure, and acute kidney disease.

The relationship between PXR activation and therapeutic effects for these diseases is described, for example, in the following literature.

Congestive liver disease special publication 2003-535912 Proc Natl Acad Sci U S A. 2001;9 8(6):3369-74.

Jaundice Hepatology. 2005; 41(3):497- 505.

Liver fibrosis Gastroenterolygy 2006;131:194-209

Acute Kidney Disease Proc Natl Acad Sci U S A. 2005;102(6):2198-203 As used herein, "hepatic fibrosis inhibition" refers to inhibiting or preventing liver fibrosis. Since activating PXR suppresses liver fibrosis, the PXR activating agent of the present invention is also useful as a preventive or therapeutic agent for liver failure.

Furthermore, since activating PXR promotes the decomposition of cholesterol, the PXR activator of the present invention can be used as a therapeutic agent for metabolic diseases such as hypertension, arteriosclerosis, hyperlipidemia, obesity, and hypercholesterolemia. (US Patent Application Publication No. 2004/0019027) The PXR activator of the present invention can also be used in combination with other drugs used for the prevention and treatment of liver diseases. The PXR activator of the present invention can be administered simultaneously with these other drugs or at different times.

[0013] The fact that the compound of general formula (I) activates PXR means that, for example, as shown in the Examples below, a PXR expression plasmid and a PXR binding sequence and a reporter gene such as luciferase are combined. Confirmed by reporter assay using a reporter plasmid. can be approved. Examples of PXR include PXR from humans and mice. Human PXR is preferred. Human PXR includes a protein having the amino acid sequence of SEQ ID NO: 2, and moreover, a protein with a total of 90% or more of the amino acid sequence of SEQ ID NO: 2. Examples include proteins that have an identity of 95% or more, and that activate a target gene in response to a ligand.

[0014] The "pharmaceutically and pharmaceutically acceptable carriers" to be blended with the compound of general formula (I), its salts, or their solvates include excipients, disintegrants, binders, and lubricants. Thickeners, coating agents, dyes, diluents, bases, tonicity agents, and the like can be used.

The dosage form of the PXR activator of the present invention is not particularly limited, but in the case of oral administration, examples include tablets, capsules, powders, fine granules, granules, liquids, syrups, etc.; Examples include injections, drips, and suppositories. Note that the PXR activator of the present invention can also be used in combination with other agents.

[0015] The administration route of the PXR activator of the present invention is not particularly limited, and it can be administered orally or parenterally. The dosage is determined as appropriate depending on the age, weight, general health condition, gender, diet, administration time, administration method, excretion rate, combination of drugs, and the severity of the patient's medical condition being treated at the time. be able to. The daily dosage varies depending on the patient's weight and condition, type of compound, administration route, etc., but for example, parenterally, approximately 0.01 to 50 mg/person administered subcutaneously, intravenously, intramuscularly, or rectally. It is desirable to administer the drug orally at a dose of about 0.01 to 150 mg/person/day, preferably 0.01 to 100 mg/person/day.

Example

[0016] The present invention will be specifically described below with reference to Examples. However, the present invention is not limited to the following examples. The compounds used as test compounds below were produced according to the method described in JP-A-6-135938. Compound 1 is Compound No. 431 of JP-A-6-135938. In Example 1 below, the fumarate salt of Compound 1 in the R form was used.

[0017] [Example 1]

PXR activation test

In HepG2 cells, which are human liver cancer-derived cells, PXR expression plasmid and CYP3A4 Transfect a reporter plasmid with luciferase linked to the expression control region of

The PXR activity of the compounds was investigated.

The PXR expression plasmid used was a pTARGET Vector (Promega) into which the human PXR gene (SEQ ID NO: 1) was inserted. The reporter plasmid was pGL3-Basic Vecror (Promega) containing XRE (xenobiotic response enhancer module: SEQ ID NO: 3 and 4) upstream of the CYP3A4 gene, dNR3 (distal nuclear receptor binding element 3: SEQ ID NO: 5), and the proximal regulatory region. (proximal pregnane phRL-TK Vector (Promega) was used as an internal standard.

HepG2 cells were cultured using 500 mL of Minimum Essential Medium (GIBCO) containing 10% FBS (fetal bovine serum), 1% sodium pyruvate, and 1% GlutaMAX (GIBCO). Cells were seeded in a 96-well plate at a concentration of 4.5 x 10 ⁴ Zwells and cultured for 24 hours. Next, the PXR expression plasmid, reporter plasmid, and internal standard plasmid were transfected using Lipofectamine 2000 (Invitrogen). After 24 hours, each concentration of test compound (DMSO solution) was added, and further incubated for 24 hours. After incubation, luciferase activity was measured using the DuaGlo Luciferase Assay System (Promega).

Table 1 and Figure 2 show the results of the target gene assay performed on rifampicin (Sigma-Aldrich Japan Co., Ltd.) and the compound 1. Both rifampicin and compound 1 activated PXR in a concentration-dependent manner, with EC50s of 0.634 M (rifampicin) and 0.084 M (compound 1), respectively. The results showed that Compound 1 activated PXR at lower concentrations than rifampicin.

[table 1]

Compound EC50 M)

Rifampicin 0.634

Compound 1 0.084 Other compounds were also examined for PXR activity and potency using reporter gene assay. As a result, the compounds shown below were also found to be highly active in PXR.

[C5]

[0020] [Example 2]

Analysis of CYP3A4 expression level by RT-PCR

Next, we investigated whether Compound 1 and rifampicin increase the amount of CYP3A4 mRNA in cultured cells.

HepG2 cells were supplemented with 0.25 M, 1 M, or 4 μM of compound 1 or 10 μM of rifampicin and cultured at 37°C for 4 days. After the culture was completed, total RNA was isolated from the cells using Trizol (Invitrogen). 1 μg of total RNA was reverse transcribed using random hexamers and SuperScriptll Transcription System (Invitrogen) to synthesize cDNA. Real-time quantitative PCR was performed using the obtained cDNA as a skeleton using a 5'-primer (SEQ ID NO: 7), a 3'-primer (SEQ ID NO: 8), and a fluorescent probe (SEQ ID NO: 9). Reaction and detection were performed using the PRISM7900 Sequence Detection System (Applied Biosystems), followed by 40 cycles of 50°C for 2 minutes, 95°C for 10 minutes, 95°C for 15 seconds, and 60°C for 1 minute. It was conducted under the following conditions. The expression level of CYP3A4 was normalized to the expression level of β-actin. The result is shown in figure 2. It was demonstrated that Compound 1 had an effect of increasing the amount of CYP3A4 mRNA to a greater extent than rifampicin.

Industrial applicability

[0021] The PXR activator of the present invention can be used to treat diseases that can be treated by activating PXR, such as liver diseases such as cholestatic liver disease and jaundice, and metabolic diseases such as hypercholesterolemia. It can be effectively used for the treatment of.

Claims

Claims: A Predanane X receptor activator comprising a compound represented by the following general formula (I), a pharmaceutically acceptable salt thereof, or a solvate thereof.

[C1]

1 4 1 4

7 9

1 4 1 alkyl group; cyclohexyl group; chel group; or C ~ C alkyl group, C ~

4 1 4 1

Alkoxy group and nozzle group of C may have one or more selected substituents.

Four

represents a strong fer group. ); or represents a cyclohexyl group,

Ring A represents a benzene ring, a thiophene ring or a furan ring.

Predanane X receptor activator according to claim 1, comprising 1-(1_cyclohexylethylamino)-4-phenylphthalazine or a pharmaceutically acceptable salt thereof, or a solvate thereof. Agent _Q

The predanane X receptor activator according to claim 1, comprising (R) -1-(1-cyclohexylethylamino)-4-phelphthalazine fumarate.

Any one of claims 1 to 3〖CYP3A4 containing the Predanane X receptor activator described herein Inducer.

[5] A drug for treating liver diseases, comprising the predanan X receptor activator according to any one of claims 1 to 3.

[6] The liver disease therapeutic agent according to claim 5, wherein the liver disease is cholestatic liver disease, jaundice, or liver failure.

[7] A liver fibrosis inhibitor comprising the predanan X receptor activator according to any one of claims 1 to 3.

[8] Any one of claims 1 to 3: A therapeutic agent for acute kidney disease comprising the predanan X receptor activator described herein.

[9] A metabolic disease therapeutic agent comprising the predanan X receptor activator according to any one of claims 1 to 3.