WO2003045433A1 - Remedies for heart failure - Google Patents

Abstract

Inhibitors (for example, a hydroxamic acid compound) for the protease activity of Adam molecules, in particular, Adam 12 are useful as remedies for heart failure in association with megalocardia.

Description

 Description Cardiovascular dysfunction drug (Technical field)

 Adam (ADAM; A Disintegrin And Metalloprotease) is a molecule containing a disintegrin-like domain containing many cystine residues and a metalloprotease-like domain, as its name suggests. It is attracting attention as a functional protein that can also exert its activity.

 The present invention relates to a heparin-binding EGF (epidermal growth factor) -like growth factor (hereinafter abbreviated as HB-EGF) by inhibiting the protease activity of this Adam molecule, more specifically, Adam 12. The present invention relates to a therapeutic agent for cardiac dysfunction associated with cardiac hypertrophy, which comprises, as an active ingredient, a compound having an action of suppressing the solubilization of the above. (Background technology)

 To date, more than 20 molecules belonging to the Adam family have been reported [for example, Adam 17 / Nature, 385, 729 (1997), ibid., 385, 733 (1997); Cell Biol., 132, 717 (1996); Adam 1 OZBiochem. J. 317, 45 (1996) ,; Adam 12 Nature, 377, 652 (1995); Adam 15 No. J. Biol. Chem., 271 , 4593 (1996); Adam 20 Gene, 206, 273 (1998), etc.].

 Many of these have been found to have transmembrane domains and to be localized at the cell membrane, playing a crucial role in molecular regulation on the cell surface. For example, Adam 17 is known as a tumor necrosis factor (TNF) convertase (solubilizing enzyme). In addition, Adam 10 is involved in the control of Notch signals and plays an important role in neurogenesis, and has also been suggested to be involved in processing of membrane proteins and degradation of extracellular matrix components. I have. It is already known that Adam 12 has protease activity, but its physiological significance is unknown.

One of the target molecules for processing by these Adam molecules is a growth factor. A variety of growth factor families are known to date, including, for example, the EGF receptor (EGFR) ligand growth factor family (EGF, HB-EGF, transforming growth factor-, amphiregulation). It is known that phosphorus, epiregulin, etc.) are synthesized in vivo as a membrane-bound form, then processed, and solubilized. For solubilization enzyme growth factors, Izumi (izumi), etc., HB EG Adam 9 solubilization process F are stated to be involved CEMB0 J., 17, 7260-7272 (1998 )] 0

Recently, growth factors have been attracting attention as one of the factors causing cardiac hypertrophy. Hypertrophy is Although it is primarily the adaptive response of the heart to various loads, the elucidation of the mechanism is important because long-term cardiac hypertrophy is also associated with chronic cardiac dysfunction and sudden death.

 It is already known that vasoconstrictive drugs such as phenylephrine, angiotensin II and endothelin-11 cause cardiac hypertrophy. All of these drugs are known to bind to G-protein coupled receptors (GPCRs) and then stimulate protein synthesis via a series of signal transductions to cause cardiac hypertrophy.

Examples in which EGFR activation is involved in a series of signal transduction systems initiated by this GPCR have been reported in various cells. For example, Eguchi et al. Have shown in an experiment using rat vascular smooth muscle cells that angiotensin II binds to GPCRs and causes EGFR phosphorylation [J. Bio. Chera. 273, 8890-8896 (1998)] ₀ Furthermore, the activation of GPCRs promoted the process of solubilizing HB-EGF from precursors, and the resulting activated HB-EGF was converted to EG. We submitted a scheme to bind and activate FR and showed that blocking the HB-EGF solubilization process could inhibit GPCR-induced EGFR activation [Nature, 402, 884] (1999)].

(Disclosure of the Invention)

 An object of the present invention is to provide a new therapeutic agent for cardiac dysfunction associated with cardiac hypertrophy. The present inventors examined the effect of a solubilizing enzyme inhibitor of HB-EGF (PCT International Publication WOO 1/70269) on the phosphorylation of EGFR in cardiomyocytes, and found that fuenorefrin, angiotensin II. It was found that phosphorylation of EGFR by endothelin-11 was inhibited, but phosphorylation by recombinant HB-EGF (solubilized HB-EGF) was not affected. Phosphorylation of EGFR by phenylephrine, angiotensin II, and endothelin-11 is also inhibited by neutralizing antibodies against HB-EGF, so that solubilized HB-EGF activates EGFR. It has been clarified that it has contributed to the development of the system. Next, the inventors identified the enzyme responsible for solubilizing HB-EGF in cardiomyocytes. It is known that the solubilization of HB-EGF is mediated by the S-form of protein kinase C (PKCS). It revealed that the enzyme was Adam 12.

 Next, the effects of a solubilizing enzyme inhibitor on cardiac hypertrophy and cardiac function were confirmed in cardiac hypertrophy model mice (see Test Examples 1 to 3 below).

From the above results, the present inventors have found that soluble HB-EGF plays an extremely important role in cardiac hypertrophy, and if this production is inhibited, cardiac dysfunction based on cardiac hypertrophy can be prevented. After confirming that it is effective for treatment, the present invention was completed. (Best mode for carrying out the invention) The therapeutic agent for cardiac dysfunction of the present invention comprises, as an active ingredient, an inhibitor for the protease activity of an Adam molecule, particularly a compound that inhibits the protease activity of Adam 12, which is a solubilizing enzyme for HB-EGF in cardiomyocytes. . The active ingredient is not particularly limited as long as it is a compound that inhibits the proteinase activity of Adam 12 in humans. For example, it is disclosed in PCT International Publication (WO 97/31109). Antagonists of meltrin (also called MeI trin) which can inhibit the protease activity of Adam 12 can be used as the therapeutic agent for cardiac dysfunction of the present invention. Many other hydroxamic acid compounds, known as matrix meta-oral protease inhibitors (MMPs), are known to inhibit the protease activity of Adam (see, for example, US Pat. 61 14361, 61 56798, 61 97810), and hydroxamic acid compounds having MMP inhibitory activity can also be used as the therapeutic agent for cardiac dysfunction of the present invention. The following compounds can also be used as the therapeutic agent for cardiac dysfunction of the present invention.

 1) 4- (N-hydroxyamino) 1-2 (R) -isobutyl-3-methylsuccinyl] _L-phenylglycine-1N-methylamide (Japanese Patent Laid-Open No. 7-101925)

 2) [4- (N-hydroxyamino) 1-2 (R) -isobutyl-3-methylsuccinyl]

-L-3- (5,6,7,8-tetrahydro-111-naphthyl) alanine-N-methylamide (PCT International Publication No. W097Z9066)

 3) [4- (N-Hydroxyamino) -1- (R) -isobutylsuccinyl]-L- 3- (1-naphthyl) alanine N-methylamide [Bioorg. Med. Chem., 5, 765-778] (1997)] 4) [4- (N-Hydroxyamino) 1-2 (R) —- ^ Γsobutyl-13- (1,2,3,4-tetrahydroisoquinolylmethyl) succinyl] -phenyl Glycine mono-N-methylamide [Bioorg. Med. Chem., 5, 765.778 (1997)]

 5) 4— (N-Hydroxyamino) -2 (R) —Isopropyl-3-methylsuccinyl] —L-Fenilalanine N-methylamide [Drug Design and Discovery, 16, 119-130 (1999)]

 6) 4- (N-hydroxyamino) 1-2 (R) -isobutyl-13-methylsuccinyl] -1-L-phenylalanine-1-N-methylamide (US Pat. No. 4,743,587)

 These compounds can be administered orally or parenterally to humans.

Oral dosage forms include solid preparations such as tablets, granules, powders, fine granules and hard capsules, and liquid preparations such as syrups and soft capsules. These preparations can be prepared in a conventional manner. Tablets, granules, powders, or fine granules may be prepared from the above compound or a pharmaceutically acceptable salt thereof, for example, lactose, starch, crystalline cellulose, superaline. It is produced by mixing with commonly used pharmaceutical additives such as magnesium acid, hydroxypropylcellulose, talc and the like. Hardness capsules are produced by filling these fine granules or powders into capsules as appropriate. The syrup is prepared by adding the above compound or its pharmaceutically acceptable solution to an aqueous solution containing sucrose, A soft capsule is prepared by dissolving or suspending an acceptable salt.A soft capsule is prepared by dissolving the compound or a pharmaceutically acceptable salt thereof in a lipid excipient, for example, a vegetable oil, an oily emulsion, or glycol. It is manufactured by suspending and filling into soft capsules.

 Examples of dosage forms for parenteral administration include injections, external preparations such as ointments, lotions and creams, suppositories such as suppositories and vaginal suppositories, and nasal administration preparations such as sprays. These preparations can be manufactured by a conventional method. For example, for an injection, the above-mentioned compound or a pharmaceutically acceptable salt thereof is dissolved in a physiological saline solution or a lipid excipient, for example, a vegetable oil, an oily emulsion, glycol, or the like. Alternatively, it is produced by emulsifying and aseptically enclosing in ampoules or vials. An ointment is prepared by a conventional method by adding the above compound or a pharmaceutically acceptable salt thereof to a base such as petrolatum, paraffin, glycerin and the like, and adding an emulsifier, a preservative, etc., if necessary. Is performed.

 The dose of the drug of the present invention varies depending on the dosage form, the age, sex or weight of the patient, or the symptom.In general, the active ingredient is 0.1 to 60 OmgZk g body weight day, preferably 10 to 20 OmgZk. The appropriate amount of g body weight per day is given once a day or divided into 2 to 4 times a day.

 The agent of the present invention significantly suppressed cardiac hypertrophy in a cardiac hypertrophy model mouse, and showed a recovery effect on cardiac function (Test Examples 1-3). Histological examination does not show any fibrosis or myofibril degradation. Therefore, the agent of the present invention is useful as a therapeutic agent for cardiac dysfunction associated with cardiac hypertrophy.

 Test example 1 Cardiac hypertrophy inhibitory effect (change in left ventricular wall thickness)

 1. Test compound

 Compound a: 4- (N-hydroxyamino) 1-2 (R) -1-isobutyl-3-methylsuccinyl] -1-L-phenylglycine-1-N-methylamide (Japanese Patent Laid-Open No. 7-110925)

2. Test method

 Using a thoracic aortic stenosis mouse (TAC mouse), a blood pressure gradient of approximately 45 Hg was created between the left and right carotid arteries at systolic pressure, and the effect on the posterior left ventricular wall thickening formed by pressure overload The effects of the test compounds were observed.

 TAC mice were C57B-6J male mice, 8 weeks old, weighing 20-25 g, and used a 27-gauge needle and suture at the site adjacent to the aorta between the left and right carotid arteries. It was prepared by stenosis.

 Compound a was suspended in 0.5% carboxymethylcellulose and administered intraperitoneally to TAC mice at 10 mg / kg per day for 4 weeks, and the thickness of the left ventricular wall was measured by echocardiography. did.

 3. Test results

Table 1 shows the measurement results. Treatment group (TAC mouse)

 Week Sham treatment group

 Compound non-administration group Compound administration group

Week 0 0.55 ± 0.27 0.54 ± 0.31 0.57 ± 0.29

After 2 weeks 0.66 ± 0.20 0.87 ± 0.30 * 0.69 ± 0.30 †

After 4 weeks 0.66 ± 0.20 1.02 ± 0.33 * 0.74 ± 0.12 †

 A significant difference test was performed on the measured values of the compound non-administration group with respect to the sham treatment group and the compound administration group with respect to the compound non-administration group, and the results were represented by * and Δ, respectively.

 * P <0.05

 † P <0.05

 The test compound clearly showed an effect of suppressing left ventricular wall thickening.

Test Example 2 Cardiac hypertrophy inhibitory effect (change in heart weight)

 1. Test compound

 Compound a: 4- (N-hydroxyamino) _2 (R) -isobutyl-13-methylsuccinyl] —L-phenylglycine-1-N-methylamide (JP-A-7-110925)

2. Test method

 In the TAC mice of Test Example 1, body weight and heart weight were measured after 4 weeks.

 (Ojo

 Table 2 shows the measurement results.

 [Table 2]

 A significant difference test was performed on the measured values of the treatment group relative to the sham-treated group and the compound administration group relative to the compound non-administration group, and the results were represented by * and そ れ ぞ れ, respectively.

 * P <0.05

 † P <O. 05

The test compound clearly showed an effect of suppressing an increase in heart weight. Test Example 3 Protective effect of cardiac function

 1. Test compound

 Compound a: 4- (N-hydroxyamino) 1-2 (R) -isobutyl-3-methylzuccinyl] -L-phenylglycine-N-methylamide (Japanese Patent Application Laid-Open No. 7-110925)

2. Test method

 Same as Test Example 1.

 After 2 weeks and 4 weeks, myocardial contractility was measured by echocardiography.

3. Test results

 Table 3 shows the measurement results.

 [Table 3]

 A significant difference test was performed on the measured values of the treatment group relative to the sham-treated group and the compound administration group relative to the compound non-administration group, and the results were represented by * and そ れ ぞ れ, respectively.

 * P <0.05

 † P <0.05

 The test compound clearly showed an effect of suppressing a decrease in myocardial contraction rate.

 Hereinafter, the present invention will be described more specifically with reference to Reference Examples and Examples.

Example 1

Tablet manufacturing

 As shown below, one tablet contains 41- (N-hydroxyamino) 1-2 (R) -1-isobutyl-3-methylsuccinyl] -1-L-phenylglycine-1-N-methylamide (compound a) l O Omg Obtain tablets.

 [Prescription]

 Component distribution Food halo

 Active agent (Compound a) 00 parts by weight

 46 parts by weight of corn starch

 98 parts by weight of microcrystalline cellulose

Hydroxypropyl cellulose Magnesium stearate

 [Operation]

Mix the base drug, corn starch and microcrystalline cellulose, add hydroxypropyl cellulose dissolved in 50 parts by weight of water, and knead well. The kneaded product is passed through a sieve, granulated on granules, and dried. The obtained granules are mixed with magnesium stearate and compressed into 25 Omg tablets.

Example 2

I notice of Ritsunari II

As shown below, a granule containing 41- (N-hydroxyamino) -2 (R) -isobutyl-13-methylsuccinyl] -1L-phenylglycine-1N-methylamide (compound a) is obtained.

 [Prescription]

 M content

Active agent (compound a) 200 parts by weight

Lactose 1 85 parts by weight

Corn starch 1 09 parts by weight

6 parts by weight of hydroxypropyl cell mouth

 [Operation]

The main drug, lactose and corn starch are mixed, and hydroxypyrucel dissolved in 120 parts by weight of water is added and kneaded well. The kneaded product is passed through a 20-mesh sieve, granulated, dried and sized to obtain a granule containing 20 Omg of the main drug in 50 Omg.

Example 3

Manufacture of capsules

As shown below, one capsule contains 100 mg of 4- (N-hydroxyamino) -2 (R) -isobutyl-1-methylsuccinyl] -one-phenylglycine-N-methylamide (Compound a) A force capsule is obtained.

 [Prescription]

 Amount

Active agent (Compound _a ) 100 parts by weight

Lactose 35 parts by weight

60 parts by weight of corn starch

Magnesium stearate

 [Operation]

The above components are thoroughly mixed, and capsules are filled with 20 mg of the mixed powder of the formula (1) to obtain capsules.

Example 4 Production of injections

4- (N-Hydroxyamino) 1-2 (R) -isobutyl-13-methylsuccinyl] -l-Phenylglycine-N-methylamide (Compound a) Injected into a mixture of 0.5 parts by weight and 5 parts by weight sorbitol Add distilled water to dissolve to make up to 100 parts by weight, and filter this aqueous solution through a membrane filter. The filtrate was filled by 5 g in ampoule was replaced with nitrogen, after溶閉obtain an injection containing the compound _a 25 m g per ampoule and sterilized for 15 minutes at 1 20 ° C.

Claims

The scope of the claims 1. A therapeutic agent for cardiac dysfunction associated with cardiac hypertrophy, which contains an inhibitor of the protease activity of the Adam molecule as an active ingredient.  2. The therapeutic agent according to claim 1, wherein the inhibitor for the protease activity of the Adam molecule is a hydroxamic acid compound having matrix metalloprotease inhibitory activity.  3. The therapeutic agent according to claim 1 or 2, wherein the Adam molecule is Adam12.