JP2003522148A - Compounds useful for treating or preventing diseases mediated by alpha-2B-adrenergic receptor - Google Patents

Abstract

  (57) [Summary] The present invention relates to the use of a selective alpha-2B-adrenergic receptor antagonist for the manufacture of a pharmaceutical preparation useful for treating or preventing a disease mediated by alpha-2B-adrenergic receptor in a mammal. The antagonist is a compound selected from the group consisting of compounds A, B, C, D and E disclosed in Scheme I, or a pharmaceutically acceptable salt of said compound. The present invention also relates to a method for preventing or treating a disease mediated by an alpha-2B-adrenergic receptor in a mammal. The method comprises administering to the mammal an effective amount of the selective alpha-2B-adrenergic receptor antagonist.

Description

Detailed Description of the Invention

The present invention relates to the use of selective alpha-2B-adrenergic receptor antagonists for the manufacture of a pharmaceutical preparation useful in the treatment or prevention of diseases mediated by alpha-2B-adrenergic receptors in mammals. The present invention also provides a method for the prevention or treatment of a disease mediated by alpha-2B-adrenergic receptor in a mammal, which comprises administering to said mammal said selective alpha-2B-adrenergic receptor antagonist. By the method.

BACKGROUND OF THE INVENTION The publications and other materials used herein to illuminate the background of the invention, and the facts to provide additional details, particularly regarding implementation, are incorporated by reference.

The selective alpha-2B-adrenergic receptor antagonists shown in Scheme I below are all known. The present inventor has found that SPECS and BioSPECS BV (
Fleminglaan, 16, 2289, CP Rijswijk, The Netherlands) from Compounds A (Order No. AE-848 / 34956037) and C (Order No. AF-399 / 36012031) and D (Order No. AH)
-034/34347043) was obtained. Compounds B (Order No. 653716) and E (Order No. 569)
063) is ChemBridge Corporation (16981 Via Tazon, Suite G, San Diego CA 921
27) provided by.

Alpha-2B-adrenergic receptors are known to mediate vasoconstriction. Thus, alpha-2B-antagonists are useful in treating or preventing diseases involving vasoconstriction. Furthermore, it has been revealed that a genetic polymorphism of the alpha-2B-adrenergic receptor gene is present in some individuals. The alpha-2B-adrenergic receptor protein of multiple subjects is a 17 amino acid acid stretch (aci) located in the third intracellular loop of the receptor polypeptide.
d strech) has a deletion of 3 glutamates from the 12 glutamate (amino acids 297-309) glutamate repeat factor (Heinonenen
et al., 1999).

OBJECTS AND SUMMARY OF THE INVENTION Compounds of the formula disclosed in Scheme I, selected from the group consisting of compounds A, B, C, D and E, are now selective alpha-2B-adrenergic receptor antagonists. It became clear that there is.

Accordingly, the present invention relates to the use of selective alpha-2B-adrenergic receptor antagonists for the manufacture of a pharmaceutical formulation useful for the treatment or prevention of diseases which mediate alpha-2B-adrenergic receptors in mammals. According to the invention, said antagonist is a compound selected from the group consisting of compounds A, B, C, D and E disclosed in scheme I, or a pharmaceutically acceptable salt of said compound.

The present invention also provides a method for the prevention or treatment of a disease mediated by an alpha-2B-adrenergic receptor in a mammal, wherein said mammal has an effective amount of a selective alpha-2B-adrenergic receptor. Comprising administering an antagonist, wherein said antagonist is a compound A, B, C as disclosed in Scheme I
, A compound selected from the group consisting of D and E, or a pharmaceutically acceptable salt of said compound.

DETAILED DESCRIPTION OF THE INVENTION Alpha-2B-adrenergic receptor antagonists are useful in the treatment and / or prevention of many diseases.

Deletion of alpha-2B-adrenergic receptor protein (Heinonen et al., 19
99), especially individuals with a deletion / deletion genotype (D / D genotype), are an important target group that would benefit from administration of selective alpha-2B-adrenergic receptor antagonists.

In a population based on the population of middle-aged Finnish men, Alpha-2B-
Those subjects with a D / D genotype of the adrenergic receptor gene were found to have a significantly increased risk for acute myocardial infarction (AMI) in a 5-year follow-up study. The risk for AMI was increased in subjects who were not previously diagnosed with coronary heart disease (CHD) at the start of the study. Therefore, the D / D genotype is
It has been postulated to be associated with impaired ability to downregulate alpha-2B-adrenergic receptor function while receptor activation is maintained. Thus, the alpha-2B-adrenergic receptor is particularly relevant in subjects with the D / D genotype, but also in subjects with I / D and I / I (I means "insertion", and " (Representing a "normal" allelic variant), may be responsible for some of the serious pathogens of all cases of AMI.

Alpha-2B-adrenoreceptor antagonists as disclosed in the present invention would be particularly useful in treating or preventing coronary heart disease. Examples include:

A) When acute AMI alpha-2B-adrenoreceptor-dependent vasoconstriction is the causative factor in some AMI cases, antagonism of these receptors restores coronary circulation and ischemic myocardium Will reduce damage.

B) Unstable angina Alpha-2B-adrenergic receptor antagonists eliminate myocardial contractile components in the discovery of persistent ischemic symptoms, thus reducing the symptoms and preventing AMI. Will.

C) Printzmetal Variant Angina Myocardial contraction is a key factor in the pathogenesis of Printzmetal angina, and alpha-2B-adrenoceptor antagonists appear to resolve and prevent seizures Be done.

D) Chronic angina and other types of CHD Alpha-2B-adrenergic receptor antagonists reduce myocardial contractile components in all CHD types, providing both symptomatic relief and protection from AMI Will. The overall reduction in vascular tone may contribute to this by reducing venous return, cardiac workload and oxygen consumption (nitrate-type effects; see below).

E) Prevention of restenosis after coronary angioplasty when myocardial contraction plays a role in restenosis Furthermore, the alpha-2B-adrenergic receptor antagonists disclosed in the present invention treat essential hypertension or It may be useful in prophylaxis, especially in subjects with increased sympathetic nerve activity and hypermuscular circulatory system.

In the above studies, D / D variants of the alpha-2B-adrenergic receptor gene have not been clearly associated with blood pressure. The inventor attributed this to two major factors, 1) antihypertensive treatment, and 2) combined control of systemic blood pressure. In another study (Heinonen et al.), It was observed that the D / D genotype was associated with reduced basal metabolic rate and reduced heart rate. These relationships probably reflect increased vascular resistance in these subjects.

In transgenic mice targeted for inactivation of the alpha-2B-adrenergic receptor gene, intravenously administered alpha-2B-adrenergic receptor agonists are found in normal animals after bolus administration of such drugs. , And fail to induce the characteristic elevation of blood pressure found in humans (Link et al., 1996). The hypotensive effects of these drugs were significantly enhanced. This demonstrates that the alpha-2B-adrenergic receptor mediates vasoconstriction. Thus, the antagonist should lower blood pressure.
This effect was not seen with alpha-2B-non-selective alpha-2B-adrenoreceptor antagonists because the antagonism of alpha-2A-adrenergic receptors was due to sympathetic outflow, cardiac output. This is to increase the volume and blood pressure. In mice with dysfunctional alpha-2A-adrenergic receptors, alpha-2-adrenergic receptor agonists elicit an enhanced hypertensive response and not hypotension (MacMillan et al., 199).
6).

Alpha-2B-adrenergic receptor antagonists are considered to have favorable effects in hypertensive subjects through their effects on renal function, muscular blood flow, and vascular resistance in other vascular beds. There is. Anti-AMI of such drug
The effect would be an additional benefit if hypertension is a significant risk factor for AMI. This protection is due to three factors: 1) lower systemic blood pressure, 2) lower risk of coronary vasoconstriction, and 3) nitrate-type effects on venous return, cardiac workload and oxygen consumption.

Further, the alpha-2B-adrenergic receptor antagonists disclosed in the present invention may be useful in treating or preventing other vascular diseases. Specifically: -hypoxic brain damage after vasoconstriction and subarachnoid hemorrhage, -migraine, -Raynaud's disease and cold intolerance, -preeclampsia, -male erectile dysfunction, and- Benefits in the treatment or prevention of obesity and metabolic syndrome can be expected.

The last-mentioned effect is due to the fact that reduced muscle blood flow and reduced basal metabolic rate contribute to the development of obesity and hypertension. Alpha-2B-adrenoceptor antagonists will increase energy expenditure by increasing muscle blood flow and favor caloric balance.

The alpha-2B-adrenergic receptor antagonists disclosed in the present invention are not selective for the alpha-2B-adrenergic receptor subtype alpha-2B.
It is also useful in analgesia and analgesia to enhance the clinical efficacy of adrenergic receptor agonists. By inhibiting the vasoconstriction induced by these agonists, co-administered alpha-2B-adrenergic receptor antagonists were only possible in the veterinary anesthesia practice and at dose levels of anesthesia not previously possible in humans. Will allow the use of larger amounts of said agonist up to.

Experimental Item Human Alpha-2-Adrenergic Receptor Binding Affinity The affinity of test compounds for the three human α ₂ -adrenergic receptor subtypes (α _2A , α _2B and α _2C ) was ³ H. -Determined in a competitive binding assay with rauwolscine. The biological material for these experiments consisted of membranes from Shionogi S115 cells stably transfected with any of the three human α ₂ subtypes (Marjamaki et al. 1992). Membrane (5-10 μg per sample
Total protein) and 1 nM to ² nM ³ H-laurorucine (specific activity 78 Ci / mmol),
Incubated with 6 concentrations of the compound in 50 mM KH ₂ PO ₄ , pH 7.5.
Each concentration was performed in duplicate. Non-specific binding was characterized by 100 μM oxymetazoline and represented 5-15% of total binding. After 30 minutes at room temperature, the incubation was stopped by high speed vacuum filtration through GF / B glass fiber filters and 3 washes with 5 ml with ice cold incubation buffer. The filtrates were then dried, impregnated with scintillate and their radioactivity determined by scintillation counting. The analysis of the experiment was carried out by fitting a non-linear least squares curve. The experimentally determined IC50 value is ch
Converted to Ki by using the eng-prusoff equation (Cheng and prusoff, 1973). The experiment was repeated a minimum of 3 times.

[Table 1]

[0024] Rat cortical alpha ₁ - binding affinity rat neocortex alpha ₁ adrenergic receptor - affinity for adrenergic receptors was determined in a competitive binding assay with ³ H- prazosin. The biological material for these assays consisted of membranes from rat neocortex. Membrane suspension (5-10 per sample
μg of total protein) and 0.2 nM to 0.25 nM ³ H-prazosin (specific activity 74 Ci / mmo)
l) is incubated with 6 concentrations of compound in a total volume of 0.25 ml (50 mM Tris, pH 7.7, 25 ° C). Each concentration was performed in duplicate. Nonspecific binding is 10μ
M is characterized by phentolamine methanesulfonate, and 25 of all bonds
It was equivalent to ~ 30%. After 30 minutes at room temperature, the incubation was performed by high-speed vacuum filtration through GF / B glass fiber filter mat and ice-cold 10 mM Tris (pH
It was stopped by 3 washes at 7.7, 4 ° C. After drying, the solid scintillates were dissolved on filter mats and their radioactivity was measured by scintillation counting.

Results At concentrations up to 30 μM, Compound A resulted in insufficient ³ H-prazosin displacement to allow estimation of IC50 values. Therefore, it was concluded that the IC50 and Ki of compound A must be above 30,000 nM.

Antagonist activity against human α ₂ -adrenoceptor subtypes The efficacy of antagonists is demonstrated in the membranes of CHO cells stably transfected with one of the three human α ₂ -adrenoceptor subtypes. , The ability of test compounds to competitively inhibit the binding of epinephrine-stimulated 35S-GTPγS to G proteins (Tian et al., 1993; Wieland and Jakobs, 1994; Jasper
et al., 1998). Membrane (2-6 μg protein per sample) and 12 concentrations of test compound were added to 50 mM Tris, 5 mM MgCl ₂ , 150 mM NaCl, 1 mM DTT, 1 mM EDTA, 10 μM GDP,
A constant concentration of epinephrine (α _2A) at room temperature in 30 μM ascorbic acid, pH 7.4.
For 5 min, 15 μM for α _2B , 5 μM for α _2C ) for 30 minutes. The binding of radiolabel was initiated by the addition of a trace amount of ³⁵ S-GTPγS (0.08M-0.15 nM, specific activity 1250 Ci / mmol) to the incubation mixture. After an additional 60 minutes at room temperature, the incubation was stopped by high speed vacuum filtration through a glass fiber filter. The filters were washed 3 times with 5 ml ice-cold wash buffer (20 mM Tris, 5 mM MgCl ₂ , 1 mM EDTA, pH 7.4 at room temperature), dried and counted in a scintillation counter for radioactivity.
The analysis of the experiment was carried out by non-linear least squares fitting. At least 3 experiments
Repeated times.

[Table 2] For purposes of the present invention, the alpha-2B-adrenoreceptor antagonists disclosed in Scheme I or pharmaceutically acceptable salts thereof can be administered by a variety of routes. Suitable dosage forms include, for example, oral formulations; parenteral injections such as intravenous, intramuscular, intradermal and subcutaneous injections; transdermal or rectal dosage forms. The required amount of alpha-2B-adrenoreceptor antagonist will vary with the particular condition being treated, the severity of the condition, the duration of treatment, the route of administration and the specific compound utilized. Suitable doses will vary from 5 μg to 100 mg per kg body weight per day and per adult.

It will be appreciated that the method of the present invention may be incorporated in various forms of embodiment, only a few of which are disclosed herein. It will be apparent to those skilled in the art that other embodiments exist and do not depart from the spirit of the invention. Therefore, the described embodiments are exemplary and should not be construed as limiting.

[Chemical 1]

[Table 3]

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 15/10 A61P 15/10 25/06 25/06 (81) Designated country EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE, TR), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, MZ, SD, SL, SZ, TZ, UG, ZW), EA (AM, AZ , BY, KG, KZ, MD, RU, TJ, TM), AE, AG, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, BZ, CA, CH, CN, CR, CU, CZ, DE, DK, DM, DZ, EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, KE, KG, KP , KR, KZ, LC, LK, LR, LS, LT, LU, LV, MA, MD, MG, MK, MN, MW, MX, MZ, NO, NZ, PL, PT, R O, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, TZ, UA, UG, US, UZ, VN, YU, ZA, ZW (72) Inventor Huobinen, Lisa Finland, Ehuen 20660 Littinen, Curly Kinkatu 5 (72) Inventor Karliokoski, Sari Finland, Ehuyen-20100 Turku, Kauppiazkatu 21 A 20 (72) Inventor Keranne, Leila Finland, Ehuyen 20760 pie Bread Heuristic, Saukonkatu 10 base 7 (72) inventor and skip, Eba - Liisa Finland, Efuien -20380 Turku, Shiutorankatu 8 F term (Reference) 4C086 BC42 GA07 MA01 MA04 NA14 ZA08 ZA36 ZA42 ZA70 ZA81

Claims (14)

[Claims] 1. A selective alpha-2 for the manufacture of a pharmaceutical preparation useful for the treatment or prevention of diseases mediated by alpha-2B-adrenergic receptors in mammals.
Use of a B-adrenergic receptor antagonist, wherein said antagonist is a compound selected from the group consisting of compounds A, B, C, D and E disclosed in Scheme I, or a pharmaceutically acceptable salt of said compound. Is used. 2. The use according to claim 1, wherein the disease is coronary heart disease (CHD). 3. The disease -Acute myocardial infarction (AMI), -Unstable angina, -Prinzmetal variant angina, -Chronic angina and other types of CHD, -Restenosis after coronary angioplasty, Use according to claim 2, wherein 4. The use according to claim 1, wherein the disease is essential hypertension. 5. The disease is Brain damage due to hypoxia after vasoconstriction and subarachnoid hemorrhage, -Migraine, -Raynaud's disease and cold intolerance, -Preeclampsia, -Male erectile dysfunction, and -Obesity, The use according to claim 1, which is a vascular disease. 6. A pharmaceutical preparation for enhancing the clinical effect of an analgesic and / or analgesic alpha-2B-adrenoceptor agonist, which is not selective for alpha-2B-adrenergic receptor subtypes. Use of a selective alpha-2B-adrenergic receptor antagonist for the manufacture of 7. A method for treating or preventing an alpha-2B-adrenergic receptor mediated disease in a mammal, said method comprising: administering to said mammal an effective amount of said selective alpha-2B-adrenergic receptor antagonist. Administering,
Wherein the antagonist comprises compounds A, B, C disclosed in Scheme I
, A compound selected from the group consisting of D and E, or a pharmaceutically acceptable salt of said compound. 8. The method of claim 7, wherein the disease is coronary heart disease (CHD). 9. The disease is -Acute myocardial infarction (AMI), -Unstable angina, -Prinzmetal variant angina, -Chronic angina and other types of CHD, -Restenosis after coronary angioplasty, The method of claim 7, wherein 10. The use according to claim 7, wherein the disease is essential hypertension. 11. The disease is Brain damage due to hypoxia after vasoconstriction and subarachnoid hemorrhage, -Migraine, -Raynaud's disease and cold intolerance, -Preeclampsia, -Male erectile dysfunction, and -Obesity, 8. The method according to claim 7, wherein the method is vascular disease. 12. An analgesic and / or analgesic alpha-2B-adrenergic receptor agonist, wherein said alpha-2B-adrenergic receptor antagonist is not selective for a subtype of alpha-2B-adrenergic receptor. 8. The method of claim 7, which is administered to a mammal to enhance the clinical effect of. 13. The 12 glutamate salts (amino acids 297-309) in a 17 amino acid stretch of acid in which the alpha-2B-adrenergic receptor antagonist is located in the third intracellular loop of the receptor polypeptide. 9. The method of claim 7, wherein the method is administered to an individual having a deletion of 3 glutamates from the glutamate repeat factor of. 14. The method of claim 13, wherein the individual is a deletion / deletion genotype.