Specification
The name of the invention: non-alcoholic fatty liver disease treatment agent
Technical field
[0001] The present invention relates to the prevention and / or therapeutic agent for nonalcoholic fatty liver disease.
Background art
[0002] As the concept of type 2 diabetes mellitus such as the so-called lifestyle-related diseases, metabolic syndrome (visceral fat syndrome) has attracted attention. Metabolic syndrome, in addition to visceral fat type obesity, hyperglycemia, hypertension is defined as a state that combines two or more any of dyslipidemia, allowed to proceed for arteriosclerosis, enhance the onset risk of myocardial infarction and cerebral infarction there is a thing.
[0003] The non-alcoholic fatty liver disease (nonalcoholic fatty liver disease, may be referred to as following NAFLD.) Is a fatty liver failure that does not depend on the drinking and obvious origin (virus and autoimmune, etc.), in recent years it has been recognized as a phenotype in liver metabolic syndrome, various etiologies of damaging fat metabolism and mitochondrial metabolism have been reported in other cases. NAFLD is, from a relatively prognosis of good simple fatty liver only by fat deposits in liver cells (simple steatosis), fibrosis of relatively severe liver tissue, cirrhosis of the liver, non-alcoholic fat that can lead to hepatocellular carcinoma sex hepatitis (nonalcoholic steatohepatitis, hereinafter sometimes referred to as NASH.) symptoms until the are encompassed (non-Patent Document 1).
[0004] as the pathogenesis of NASH, of Day et al., "Two hit theory" is widely known (Non-Patent Document 2). In other words, in the process the formation of fatty liver (1st hit), imbalance and of caloric intake and consumption balance, lipid storage of insulin resistance to liver cells by metabolic abnormalities that the foundation is involved. Fatty liver proceeds to NASH as further 2nd hit, activation of growth and the innate immune system associated therewith oxidative stress based on energy metabolism load plays an important role. Kupffer (CD68-positive) cells are immunocompetent cells in liver are resident macrophages of the liver, it has been reported to be increased in NASH patients (Non-Patent Document 3). Furthermore even in the livers of NASH model mice, it has been reported that macrophages are increasing (Non-Patent Document 4). By removing the experimentally Kupffer cells, the fat accumulation in liver induced by high-fat diet is suppressed also been reported, suggesting that Kupffer cells play an important role in the pathogenesis of NASH is (non-Patent Document 5).
[0005] The treatment of NASH is, obesity, diabetes, dyslipidemia, living habits improve the diet and exercise therapy was mainly for lifestyle-related diseases such as high blood pressure the underlying. However, in practice the improvement of lifestyle is difficult, it is believed to be an important factor in the progression of NASH, insulin resistance, oxidative stress, dyslipidemia, been carried out medications that target hypertension, etc. there. The insulin resistance improving drug, ligand and a thiazolidinedione derivative of the nuclear receptor PPARγ involved in enhancing action of insulin sensitivity (pioglitazone and rosiglitazone, etc.), or, of insulin resistance improving drug biguanide drugs (metformin, etc.) but as the antioxidant, vitamin E have been used as therapeutic agents by combination with either alone or vitamin C. In addition, as the abnormal lipid metabolism of therapeutic agents, PPARα agonist is a fibrate drug (fenofibrate and bezafibrate, etc.), statin formulation, probucol, etc., as the hypertension therapeutic agents, angiotensin II type 1 receptor antagonist (ARB ) it is expected as each therapeutic agent. In particular, in the fibrate and statin drugs are expected in terms of anti-inflammatory action with their medication. However, such as the improvement of symptoms in fatty liver failure, high reported less level of evidence, highly recommended degree of treatment has not been established. By has continued to increase in the affected individuals worldwide metabolic syndrome, it is expected to increase in the future as well the number of patients of NASH, establishment of treatment is desired (Non-Patent Document 1, 6).
[0006] On the other hand, Patent Document 1, the following equation (1):
[0007] [Formula 1]

[0008] (In the formula, R1 and R2 the same or different and each represents a hydrogen atom, a methyl group or an ethyl group; R3a, R3b, R4a and R4b are the same or different and each represents a hydrogen atom, a halogen atom, a nitro group, a hydroxyl group, C1 -4 alkyl group, a trifluoromethyl group, C1-4 alkoxy group, C1-4 alkylcarbonyloxy group, di -C1-4 alkylamino group, C1-4 alkylsulfonyloxy group, C1-4 alkylsulfonyl group, C1- 4 alkylsulfinyl group, or a C1-4 either indicating the alkylthio group, an alkylenedioxy group bonded R3a and R3b, or R4a and R 4 b; X-represents an oxygen atom, a sulfur atom or N-R5 (R5 is a hydrogen atom, . which C1-4 alkyl group, C1-4 alkylsulfonyl group, a C1-4 alkyloxycarbonyl group) indicates; Y is an oxygen atom, S (O) l group (l represents a number of 0 to 2). , shows a carbonyl group, a carbonyl amino group, amino carbonyl group, sulfonylamino group, amino sulfonyl group, or an NH group; Z represents CH or N; n represents a number of 1 ~ 6; m is 2 to 6 It indicates the number.)
The compound represented by [0009] These salts or solvate thereof has a selective PPARα activating action, without the weight gain and obesity in mammals, including humans, hyperlipidemia , arteriosclerosis, diabetes, diabetic complications (diabetic nephropathy, etc.), inflammation, it is disclosed that is useful as a preventive and / or therapeutic agent, such as heart disease. However, these compounds NAFLD, there is no mention or suggest, especially for or what kind of action to NASH symptoms were severe.
Prior art documents
Patent literature
[0010] Patent Document 1: International Publication No. 2005/023777 pamphlet

Non-patent literature
[0011] Non-Patent Document 1:...... Nugent C. et al, Nat Clin Pract Gastroenterol Hepatol, 4 (8), 432-41 (2007)
Non-Patent Document 2:. Day CP et al, Gastroenterology, 114 (4), 842-5 (1998)
Non-Patent Document 3:.... Park JW et al, J. Gastroenterol Hepatol, 22, 491-7 (2007)
Non-Patent Document 4:.... Yoshimatsu M. et al, Int J. Exp Path, 85, 335-43 (2004)
Non-Patent Document 5:. Stienstra R. et al, Hepatology, 51 (2), 511-22 (2010)
Non-Patent Document 6: Hirofumi Uto other, the latest medicine, Vol. 63, No. 9, 1683-7 (2008)

Summary of the invention
Problems that the Invention is to Solve
[0012] The world has continued to increase in the sufferers of metabolic syndrome, NAFLD, in particular, but the symptoms are expected to increase in the future as well the number of patients of NASH was severe, the level of evidence is with respect to the treatment of NAFLD high report small, high therapeutic methods of the recommended size is at present not been established. The object of the present invention is to provide NAFLD, especially the prevention of NASH, prevention useful new non-alcoholic fatty liver disease treatment, a therapeutic agent.
Means for Solving the Problems
[0013] The present inventors have found that non-alcoholic fatty liver disease (NAFLD), in particular the prevention of the symptoms of severe non-alcoholic steatohepatitis (NASH), to find a compound useful in the treatment, in NASH model animal some LDL receptor knockout mice and MCD (methionine - choline deficiency) that prevented using the food load KK-Ay mice, was subjected to a search of the compound useful in the treatment, example 85 in the patent literature 1 is also in quite surprising compounds disclosed as is, to suppress the ballooning swelling and fat deposition in liver cells, they found that reducing the Kupffer cells, prevention of NAFLD found that compound useful in the treatment, completed the present invention did.
[0014] Namely, the present invention is, (R) -2- [3 - [[N- (benzoxazol-2-yl) -N-3- (4- methoxyphenoxy) propyl] aminomethyl] phenoxy] butyric acid ( hereinafter, sometimes referred to as compound the a.), or is to provide a preventive and / or therapeutic agent for nonalcoholic fatty liver disease and its salts or active ingredient solvate thereof.
[0015] When explaining the present invention in more detail, relates to the present invention is the following (1) (18).
(1) (R) -2- [3 - [[N- (benzoxazol-2-yl) -N-3- (4- methoxyphenoxy) propyl] aminomethyl] phenoxy] butyric acid, or a salt or thereof prevention and / or therapeutic agent for nonalcoholic fatty liver disease as an active ingredient solvate.
(2) non-alcoholic fatty liver disease is non-alcoholic steatohepatitis, prevention and / or therapeutic agent according to the above (1).
(3) prevention and / or treatment of non-alcoholic fatty liver disease, is to suppress the fat deposition in the liver, preventing and / or therapeutic agent according to (1) or (2).
(4) the prevention and / or treatment of non-alcoholic fatty liver disease, is intended to reduce the Kupffer cells in the liver, the prevention and / or therapeutic agent according to the above (1) or (2).
[0016] (5) The compound A, or inhibiting or reducing agent of fat deposition in the liver to its salt or active ingredient these solvate.
[0017] (6) The compound A, or inhibiting or reducing agent of Kupffer cells in the liver to its salt or active ingredient these solvate.
[0018] (7) the compound to a patient in need A, or which comprises administering an effective amount of a salt or solvate thereof, preventing and / or treating non-alcoholic fatty liver disease.
(8) non-alcoholic fatty liver disease is nonalcoholic steatohepatitis, a method according to (7).
(9) to a patient in need suppression or reduction of fat deposits in the liver, which comprises administering said compound A, or an effective amount of a salt or solvate thereof, inhibit fat deposition in the liver or how to decrease.
(10) patient in need increased inhibition or decrease in the Kupffer cells in the liver, the compound A, or which comprises administering an effective amount of a salt or solvate thereof, the Kupffer cells in the liver how to suppress or decrease.
[0019] (11) for the manufacture of a pharmaceutical composition for preventing and / or treating non-alcoholic fatty liver disease, said compound A, or the use of a salt or solvate thereof.
(12) nonalcoholic fatty liver disease is nonalcoholic steatohepatitis, used according to (11).
(13) for the manufacture of a pharmaceutical composition for inhibiting or reducing fat deposition in the liver, the compound A, or the use of a salt or solvate thereof.
(14) for the manufacture of a pharmaceutical composition for inhibiting or reducing the Kupffer cells in the liver, the compound A, or the use of a salt or solvate thereof.
[0020] (15) for use in pharmaceutical compositions used to prevent and / or treat non-alcoholic fatty liver disease, said compound A, or a salt or solvate thereof.
(16) for use in the pharmaceutical compositions used to prevent and / or treat non-alcoholic steatohepatitis, the compound A, or a salt or solvate thereof.
(17) for use in pharmaceutical compositions which are used to inhibit and / or reduce fat deposits in the liver, the compound A, or a salt or solvate thereof.
(18) for use in pharmaceutical compositions which are used to Kupffer cells is inhibited and / or reduced in the liver, the compound A, or a salt or solvate thereof.
Effect of the invention
[0021] The present invention, non-alcoholic fatty liver disease (NAFLD), and in particular, to provide a new agent for the prophylaxis or treatment of nonalcoholic steatohepatitis severe (NASH). Especially for NASH is severe, fewer reports of a high level of evidence the therapeutic agent, highly recommended degree of treatment has not been established, and it is expected that the number of patients will increase in the future in the world, treatment establishment of law has been desired. The present invention suppresses the balloon-like swelling of the fatty deposits or liver cells of liver, it is possible to further reduce the Kupffer cells in the liver, for the more severe NASH, it is provided a prophylactic or therapeutic agent is there.
Brief description of the drawings
[0022] [FIG 1] FIG 1 shows Compound A (0.5mg / kg) or fenofibrate (100mg / kg) balloon-like swelling of hepatocytes when administered according to the present invention (Ballooning of hepatocytes) it is a diagram.
FIG. 2 is a view showing a compound of the present invention A (0.5mg / kg) or fenofibrate (100mg / kg) CD68 positive cell area ratio in the liver when administered (CD68 positive area in liver) it is.
[3] FIG 3 is a diagram illustrating a fatty liver score (Steatosis score) when compounds were administered A (0.25mg / kg) or bezafibrate (60mg / kg) of the present invention.

DESCRIPTION OF THE INVENTION
[0023] used in the present invention (R) -2- [3 - [[N- (benzoxazol-2-yl) -N-3- (4- methoxyphenoxy) propyl] aminomethyl] phenoxy] butyric acid (Compound A ) is, for example, may be prepared as described in WO2005 / 023 777 pamphlet or the like. It is also possible to formulated according to the method described in the literature.
[0024] Further, the present invention can also be used salts or solvates of the compounds the A. Salt and solvent mixture can be by a conventional method, to manufacture.
[0025] As a salt of Compound A, is not particularly limited as long as pharmaceutically acceptable, for example, sodium salts, alkali metal salts such as potassium salts; calcium salts, alkaline earth metal salts such as magnesium salt ; ammonium salts, organic base salts such as trialkylamine salts; hydrochloride, mineral such as sulfuric acid salts; and an organic acid salt such as acetate and the like.
[0026] Compound A, or a solvate of a salt thereof, a hydrate, alcohol solvate (e.g., ethanol solvate) and the like.
[0027] The compound represented by the Compound A is an optical isomer of R-form and S-form because it has an asymmetric carbon atom are present, they are all encompassed in the present invention.
[0028] In NASH, (see Non-Patent Document 3) are known to macrophages in a CD68-positive Kupffer cells is increased in the liver. As shown in the following example, Compound A is a NASH animal model, in the high-fat, high-cholesterol diet (Western diet) load LDL receptor knockout mice, a significant decrease of balloon-like swelling and Kupffer cells of the liver cells the shows, in MCD diet KK-Ay mice, a further NASH animal model, showed a significant reduction of fat deposition in the liver. Thus, compound A, or a salt thereof, or solvate thereof, of the present invention, the prevention and / or treatment of non-alcoholic fatty liver disease, especially high severity non-alcoholic steatohepatitis of mammals, including humans it is useful as an agent.
[0029] Prevention and / or therapeutic agent of the present invention, the active ingredient and a pharmaceutical composition comprising a pharmaceutically acceptable carrier for the prevention, the active ingredient for the treatment and a pharmaceutically acceptable carrier It encompasses a pharmaceutical composition comprising, as well as prevention and the active ingredient and a pharmaceutical composition comprising a pharmaceutically acceptable carrier for therapy.
Fat deposition inhibitor in the liver of the present invention, decreases the pharmaceutical composition containing the active ingredient and a pharmaceutically acceptable carrier for to suppress the increase in the deposition of fat in the liver, fat deposits in the liver active ingredients and pharmaceutical compositions comprising a pharmaceutically acceptable carrier for causing, as well as contain the active ingredient and a pharmaceutically acceptable carrier for causing the suppression and decrease the increased deposition of fat in the liver It encompasses the a pharmaceutical composition.
Kupffer cell reduction agent in the liver of the present invention, a pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable carrier in order to suppress an increase in the Kupffer cells in the liver, Kupffer cells increased in the liver active ingredients and pharmaceutical compositions comprising a pharmaceutically acceptable carrier for reducing, and active ingredients for reducing the Kupffer cells increased in suppression of increase in the Kupffer cells in the liver and the liver and a pharmaceutically It encompasses a pharmaceutical composition comprising an acceptable carrier.
Prevention and / or therapeutic agents, and pharmaceutical compositions of the present invention of the present invention is a compound represented by the Compound A, or is intended to its salt, or a solvate thereof as an active ingredient, an effective of the present invention component, as shown in examples described later, is useful as a preventive and / or therapeutic agent for highly non-alcoholic fatty liver disease, in particular the severity of mammalian non-alcoholic steatohepatitis, including humans. Further, as shown in examples described later, the active ingredient of the present invention, fat deposition inhibitor in the liver of mammals, including humans, are also useful as Kupffer cells reducing agent in the liver.
[0030] Compound A, or a salt or solvate thereof of the present invention using a single, or other pharmaceutically acceptable carriers as tablets, capsules, granules, powders, lotions, ointments , injections can be a dosage form such as a suppository. These preparations can be produced by a known method. For example, in the case of a preparation for oral administration, tragacanth, gum arabic, sucrose fatty acid esters, lecithin, olive oil, soybean oil, dissolving agent of PEG400 and the like; starch, mannitol, an excipient such as lactose; carboxymethylcellulose sodium, binders such as hydroxypropylcellulose; disintegrating agents such as crystalline cellulose, carboxymethylcellulose calcium; prepared as talc, lubricants such as magnesium stearate, by formulating appropriately combined flow improver such as light anhydrous silicic acid and the like can do.
[0031] Compound A, or a salt or solvate thereof of the present invention are administered by oral or parenteral administration. The dosage of the pharmaceutical of the present invention, the patient's weight, age, sex, varies according to the symptoms and the like, in the case of adult, daily 0.01 - 1000 mg as the compound A, preferably having from 1 to 0.1 - 100 mg 3 preferably administered in time.
EXAMPLE [0032] Hereinafter, the present invention will be described more specifically with reference to Examples, the present invention is not limited in any way by these examples.
[0033] effects in Example 1 LDL receptor knockout mice NASH model
NASH may develop inflammation of fatty liver and liver are characteristic condition are known, and examined the effects of Western diet loading LDL receptor knockout mice (Non-Patent Document 4).
Incidentally, in this study, the disclosed as Example 85 in Patent Document 1, (R) -2- [3 - [[N- (benzoxazol-2-yl) -N-3- (4- methoxyphenoxy) propyl] aminomethyl] phenoxy] butyric acid (compound a) was used.
[0034] 1) Animals used:
Male LDL receptor knockout mice (B6.129S7-Ldlr / J, Jackson Laboratories) was used for the experiment. 13 weeks from 8 weeks of age before and after, Teklad Custom Research Diet (Harlan Teklad Inc., TD.88137) as a Western diet ad libitum and allowed to develop NASH.
2) group configuration:
The Western diet loaded onto a mouse, were carried out blood sampling and measurement of body weight after one week. As there is no difference in lipid and weight in plasma between groups, the control group (0.5% aqueous methyl cellulose solution), Compound A 0.5mg / kg dose group, and were grouped into fenofibrate 100 mg / kg administered group.
3) drug administration:
The dose volume was set to 5 mL / kg body weight, 0.5% methylcellulose aqueous solution for the control group, Compound A administration group, each of the chemical liquid in the fenofibrate administration group was orally administered once a day. The administration period was 12 weeks.
[0035] 4) observation and inspection method:
After completion of administration, the liver was removed under pentobarbital sodium (50mg / kg) anesthesia, were fixed with paraformaldehyde, hematoxylin - were prepared immunostained specimens against eosin stained specimen and CD68. Hematoxylin - by using the eosin stained specimen, a balloon-like swelling of the liver cells in blind conditions, the following criteria (. Kleiner et al Hepatology 41, 1313-21, 2005) were scored in.
Balloon-like tumor large cell No: 0
Balloon-like tumor large cell is a slight (Few balloon cells): 1
Balloon-like tumor large cell is more or remarkable (Ballooning): 2
CD68-positive cells ratio of the area of ​​the liver to the blind under the (CD68 positive area in liver) was measured by an image analysis system (WinROOF).
Statistical processing was performed using the Stat Preclinica ((Ltd.) Takumi Information Technology) (Ver.1.1). The Dunnett's test (N = 14-15), *** to the control group, the * mark in the case of a significant difference at p-<0.05, if there is a significant difference p <0.001 It marked with.
[0036] 5) Result:
As shown in FIG. 1, the 0.5 mg / kg administration group of Compound A, significant inhibition of balloon-like swelling of the hepatocytes was observed. In the 100mg / kg dose group of fenofibrate, they tend to suppress the balloon-like swelling, but was found was not statistically significant. Further, as shown in FIG. 2, the 0.5 mg / kg dose group of compound the A, CD68 positive cells area ratio in the liver was reduced markedly (reduction rate 81%). Although lowering of CD68-positive cells ratio of the area of ​​the liver in the 100mg / kg dose group of fenofibrate was observed, the reduction rate is 73%, did not reach to the effect of the compound A administration group. These results, Compound A is a pathology of NASH, positive area of ​​the balloon-like swelling and Kupffer cells of the liver cells, was found to potently inhibit than fenofibrate is PPARα agonist.
[0037] action in NASH model using a second embodiment of methionine and choline-deficient diet KK-Ay mouse
By loading methionine choline deficient diet (MCD diet) in experimental animals, it is known to develop fatty liver which is a characteristic pathology of NASH, the MCD diet-fed the KK-Ay mice (Nakano S. et al ., Hepatol Res., 38 (10), were examined effects in 1026-39,2008). Incidentally, in this study, Example 1 and is disclosed as Example 85 in Patent Document 1 as well, (R) -2- [3 - [[N- (benzoxazol-2-yl) -N 3 was used (4-methoxyphenoxy) propyl] aminomethyl] phenoxy] butyric acid (compound a).
[0038] 1) Animals used:
Male KK-Ay mice (KK-Ay / TaJcl, CLEA Japan, Inc.) were used for the experiment. 12-week-old ad libitum for 16 weeks MCD diet than before and after, was the onset of NASH.
2) group configuration:
As there is no difference in body weight between the groups, normal diet group, the control group (MCD diet), Compound A 0.25mg / kg administered group, and were grouped into bezafibrate 60 mg / kg administered group.
3) drug administration:
The diet was carried out by the administration. The MCD diet containing no drug control group, the compound A administration group with MCD diet containing the compound A 0.00026%, the bezafibrate administration group were allowed to freely take MCD diet containing 0.06% of bezafibrate. The administration period was 16 weeks.
[0039] 4) observation and inspection method:
After completion of administration, the liver was removed under pentobarbital sodium (50mg / kg) anesthesia, were fixed with paraformaldehyde, hematoxylin - were prepared eosin stained specimen. In blind conditions to evaluate the fatty liver score (Steatosis score). Fat degree of deposition (for Grade) was observed at a magnification 100 were scored from 0 to 3 fatty liver to the following criteria depending on the degree.
Less than 5%: 0
Less than 5% or more and 33%: 1
Less than 33% 66% or more: 2
66% or more: 3
Statistical processing was performed using the Stat Preclinica ((Ltd.) Takumi Information Technology) (Ver.1.1). The Dunnett's test (N = 4-9), *** to the control group, the * mark in the case of a significant difference at p-<0.05, if there is a significant difference p <0.001 It marked with.
[0040] 5) Result:
As shown in FIG. 3, in 0.25mg / kg administered group of compound A, fatty liver score is 0 in all cases, ie fat deposition has disappeared almost completely. In the 60mg / kg dose group of bezafibrate, but suppressed the fat deposition to the same extent as the normal diet group, it did not reach until almost completely fat deposits as of Compound A is lost. From these things, Compound A fat deposits, one of NASH disease states, was found to potently inhibit than bezafibrate is PPARα agonist.
[0041] or more, from Example 1 and Example 2, Compound A of the present invention is extremely becomes clear that is useful as a preventive or therapeutic drug for non-alcoholic steatohepatitis (NASH), nonalcoholic fatty liver it is extremely useful as a preventive or therapeutic drug for a disease (NAFLD) revealed.
Industrial applicability
[0042] The present invention provides a compound A, or its salt or solvate thereof, severe hepatic cells in non-alcoholic steatohepatitis (NASH), which is the pathology of nonalcoholic fatty liver disease (NAFLD) balloon-like swelling of the suppression, the positive area reduction of Kupffer cells, found for the first time that it has an inhibitory effect of fat deposition, provides a preventive and / or therapeutic agent for low-molecular-weight non-alcoholic fatty liver disease it is intended. The present invention is useful in the pharmaceutical industry since it is available as a pharmaceutical drug substance, and has industrial applicability.
The scope of the claims
[Claim 1] (R) -2- [3 - [[N- (benzoxazol-2-yl) -N-3- (4- methoxyphenoxy) propyl] aminomethyl] phenoxy] butyric acid, or a salt thereof, or prevention and / or therapeutic agent for nonalcoholic fatty liver disease to these solvate as an active ingredient.