CN1450995A - 5-hydroxy indazole derivatives for treating glaucoma - Google Patents

Abstract

3-(2-aminoethyl)-1H-indazol-5-ols useful for treating elevated intraocular pressure and glaucoma are disclosed.

Description

5-Hydroxyindazole derivatives for the treatment of glaucoma

The present invention relates to substituted 3-(2-aminoethyl)-1H-indazole-5-phenols, certain substituted 3-(2-aminoethyl)-1H-indazole-5-phenols are novel for the reduction and control of normal or elevated intraocular pressure (IOP) and the treatment of glaucoma.

Background of the invention

The disease known as glaucoma is characterized by permanent loss of visual function due to irreversible damage to the optic nerve. Several morphologically and functionally distinct types of glaucoma are generally characterized by elevated IOP, which is thought to be causally related to the pathological process of the disease. Ocular hypertension is a disorder in which intraocular pressure is elevated without significant loss of visual function; such patients are considered to be at high risk for eventual development of glaucoma-related visual loss. Some patients with glaucomatous visual field loss have relatively low intraocular pressure. These so-called normotensive or hypotensive glaucoma patients may also benefit from agents that lower or control IOP. Loss of visual function, or its progressive deterioration, generally improves when glaucoma or ocular hypertension is detected early and treated promptly with drugs that effectively lower intraocular pressure. Drug therapies that have proven effective in reducing intraocular pressure include agents that reduce aqueous humor production and agents that promote patency of outflow. Such treatments are generally administered by one of two possible routes, topically (direct application to the eye) or orally.

Some individuals do not respond well when treated with certain existing glaucoma treatments. Therefore, there is a need for other topical therapeutic agents to control IOP.

Serotonergic compounds having 5- _HT2 receptor agonist activity have now been found to be effective in lowering and controlling normal and elevated IOP and are useful in the treatment of glaucoma, see co-owned co-pending application PCT/US99 /19888. Compounds that are agonists of the 5- _HT2 receptor are well known and have shown various utility, primarily for diseases or conditions associated with the central nervous system (CNS). US Patent (US Patent) 5,494,928 discloses certain 2-(indol-1-yl)-ethylamine derivatives as 5-HT _2C agonists for the treatment of obsessive-compulsive disorder and other personality disorders of CNS origin . U.S. Patent No. 5,571,833 discloses tryptamine derivatives as 5-HT ₂ agonists for the treatment of portal hypertension and migraine. Methods of treating malaria using 5-HT _2A/2C agonists are disclosed in US Patent 5,874,477. US Patent No. 5,902,815 discloses the use of 5-HT _2A agonists to prevent the adverse effects of NMDA receptor hypofunction. 5-HT _2B agonists are disclosed in WO98/31354 A2 for the treatment of depression and other CNS diseases. Response of 5-HT _2A receptors to agonists has been reported to be the major activity responsible for hallucinogenic activity, while some lesser 5-HT _2C receptors may be involved [Psychopharmacology, Vol. 121: 357 , 1995].

The present invention relates to substituted 3-(2-aminoethyl)-1H-indazole-5-phenols, certain substituted 3-(2-aminoethyl)-1H-indazole-5-phenols are novel of. These compounds have a high affinity with the 5- _HT2 receptor of serotonin and act as agonists of the latter, so they are effective in reducing and controlling normal or elevated intraocular pressure (IOP) and glaucoma. Treatment can be helpful. When a phenolic moiety is included in this substitution, for example when there is a hydroxyl group at the 5-position of the indazole ring, such compounds are susceptible to the well-known Oxidation reactions are particularly sensitive [Journal of Physical Chemistry (J.Phys.Chem.), 103,8606 (1999), Chemical Research and Toxicology (Chem.Res.Toxicol.), 11, 639 (1998), Journal of Organic Chemistry (J . Org. Chem.), 52, 2817 (1987), J. Pharm. Sci, 77, 911 (1988)], which are particularly relevant to the present application. Such hydroxy-substituted phenols can be protected from oxidation by derivatization of the aryl hydroxy groups to generate appropriate esters (carbamate or carbonate). Although esters (carbamate or carbonate derivatives) themselves do not have high affinity for the above-mentioned receptors, they are indeed useful in the treatment of glaucoma because properly protected phenols can be activated in vivo by chemical hydrolysis or tissue esterification. Decomposed by the action of enzymes. Such decomposition will release the desired therapeutic agent, the desired novel 5-hydroxy-indazole compounds disclosed in this invention. The concept of using such derivatized phenolic compounds as chemical delivery agents is well known in the art [Drugs Pharm. Sci., 53, 221 (1992), Pharm. Res., 168 (1984)].

The chemical synthesis of 3-(2-dimethylamino-ethyl)-1H-indazol-5-ol has been reported, but the utility of this compound has not been described [J.Amer.Chem.Soc .), 79, 5242 (1957); Journal of the American Chemical Society, 80, 965 (1958)].

The chemical synthesis of 1-(2-aminopropyl)-1H-indazol-6-ol and other ring substitution variants has been reported in published International Patent Application WO 98/30548 (1998). The listed utility of the compounds of this application is for the treatment of diseases of the central nervous system such as disease, reproductive insufficiency, appetite regulation disorders, anxiety, depression and sleep disorders. This application does not mention the use in ophthalmology. Published International Patent Application WO 00/12482 (2000) discloses certain 1-(indazol-3-yl)-2-propanamine derivatives as 5-HT ₂ agonists for use in the treatment of central nervous system disorders.

Summary of the invention

The present invention relates to derivatives of 3-(2-aminoethyl)-1H-indazol-5-ol, some of which are novel, and which are useful for reducing and controlling hypertensive disorders in warm-blooded animals, including humans. Glaucoma, ocular hypertension, and glaucoma-related IOP. These compounds are formulated into pharmaceutical compositions suitable for topical ocular use. Description of the preferred embodiment

The following formula I and pharmaceutically acceptable salts and solvates of compounds of formula I represent useful compounds of the present invention. Structural Formula I

Wherein G is selected from hydrogen, halogen or C _1-4 alkyl;

R is hydrogen, C _1-4 alkyl, C(=O)W or P(=O)(OX)(OY),

R ^{and R} are hydrogen;

R ³ and R ⁴ are independently selected from hydrogen, C _1-4 alkyl, or R ³ , R ⁴ form a cyclopropyl ring together with the carbon atoms they are attached to, or in addition, R ² and R ³ together form (CH ₂ ) _m to form a saturated heterocycle;

When R ² and R ³ are part of a heterocycle, R ¹ can be hydrogen or C _1-4 alkyl;

^R can be hydrogen or C _1-4 alkyl;

When R, R ¹ , R ² , R ⁵ and G are all hydrogen, R ³ and R ⁴ cannot be hydrogen at the same time;

W is C _1-6 alkyl, NR ⁶ R ⁷ , N(R ⁶ )CH ₂ (CH ₂ ) _n C(=O)N(R ⁷ )(R ⁸ ), OC _1-6 alkyl, C _{1 -6} alkyl (which can be replaced by halogen, hydroxyl, CO ₂ C _1-4 alkyl, CON(C _1-4 alkyl) ₂ , C(=NH)NH ₂ , HC(=NH)NH ₂ , NH ₂ substituted), C _2-4 alkenyl (substituted by phenyl, not or substituted by one or more of C _1-4 alkyl, C _1-4 alkoxy or halogen);

R ⁶ , R ⁷ and R ⁸ are independently selected from hydrogen or C _1-4 alkyl;

X and Y are independently selected from hydrogen, C _1-10 alkyl, or X and Y can jointly form a lower (CH ₂ ) _m alkyl chain;

m is 2-4;

n is 1 or 2.

The novel and useful compounds of the present invention can be defined as follows:

G is selected from hydrogen, halogen or C _1-4 alkyl;

R is C(=O)W or P(=O)(OX)(OY),

R ^{and R} are hydrogen;

R ³ , R ⁴ form a cyclopropyl ring together with the carbon atoms they are connected to, or in addition, R ² and R ³ jointly form (CH ₂ ) _m to form a saturated heterocyclic ring;

When R ² and R ³ are part of a heterocycle, R ¹ can be hydrogen or C _1-4 alkyl;

^R can be hydrogen or C _1-4 alkyl;

W is C _1-6 alkyl, NR ⁶ R ⁷ , N(R ⁶ )CH ₂ (CH ₂ ) _n C(=O)N(R ⁷ )(R ⁸ ), OC _1-6 alkyl, C _{1 -6} alkyl (which can be replaced by halogen, hydroxyl, CO ₂ C _1-4 alkyl, CON(C _1-4 alkyl) ₂ , C(=NH)NH ₂ , HC(=NH)NH ₂ , NH ₂ substituted), C _2-4 alkenyl (substituted by phenyl, not or substituted by one or more of C _1-4 alkyl, C _1-4 alkoxy or halogen);

R ⁶ , R ⁷ and R ⁸ are independently selected from hydrogen or C _1-4 alkyl;

X and Y are independently selected from hydrogen, C _1-10 alkyl, or X and Y can jointly form a lower (CH ₂ ) _m alkyl chain;

m is 2-4;

n is 1 or 2.

Preferred compounds are:

G is selected from hydrogen, halogen or C _1-4 alkyl;

R is hydrogen, C(=O)W or P(=O)(OX)(OY),

R ^{and R} are hydrogen;

R ³ and R ⁴ are independently selected from hydrogen, C _1-4 alkyl, or R ³ , R ⁴ form a cyclopropyl ring together with the carbon atoms they are connected to;

^R can be hydrogen or C _1-4 alkyl;

W is C _1-6 alkyl, NR ⁶ R ⁷ , N(R ⁶ )CH ₂ (CH ₂ ) _n C(=O)N(R ⁷ )(R ⁸ ), C _1-6 alkyl (which can be Substituted by halogen, hydroxyl, CO ₂ C _1-4 alkyl, CON(C _1-4 alkyl) ₂ , C(=NH)NH ₂ , HC(=NH)NH ₂ , NH ₂ ), C _2-4 Alkenyl (substituted by phenyl, not or substituted by one or more of C _1-4 alkyl, C _1-4 alkoxy or halogen);

R ⁶ , R ⁷ and R ⁸ are independently selected from hydrogen or C _1-4 alkyl;

X and Y are independently selected from hydrogen, C _1-10 alkyl, or X and Y can jointly form a lower (CH ₂ ) _m alkyl chain;

m is 2 or 3;

n is 1 or 2.

The most preferred compounds are:

G is selected from hydrogen, halogen or C _1-4 alkyl;

R is hydrogen or C(=O)W;

R ^{and R} are hydrogen;

R ³ is hydrogen, and R ⁴ is methyl, or R ³ and R ⁴ form a cyclopropyl ring together with the carbon atoms to which they are attached;

^R is hydrogen;

W is C _1-6 alkyl, C _1-6 alkyl (which may be substituted by halogen, hydroxyl, CON(C _1-4 alkyl) ₂ , C(=NH)NH ₂ ).

Representative examples of preferred novel compounds of formula I are:

3-(2-Aminopropyl)-1H-indazol-5-ol;

3-(2-Aminopropyl)-1-methyl-1H-indazol-5-ol;

2-(5-methoxy-1H-indazol-3-yl)-1-methyl-ethylamine;

3-(2-Aminopropyl)-6-fluoro-1H-indazol-5-ol;

3-(2-Aminopropyl)-7-methyl-1H-indazol-5-ol;

3-(2-Aminopropyl)-6-fluoro-1-methyl-1H-indazol-5-ol;

2-Methyl-propionic acid 3-(2-aminopropyl)-1H-indazol-5-yl ester;

2,2-Dimethyl-propionic acid 3-(2-aminopropyl)-1H-indazol-5-yl ester;

3-(2-aminopropyl)-1H-indazol-5-yl cyclopropanecarboxylate;

N,N-diethyl-succinamic acid 3-(2-aminopropyl)-1H-indazol-5-yl ester;

It is recognized that compounds of formula I may contain one or more chiral centers. The present invention contemplates all enantiomers, diastereomers and mixtures thereof.

In the above definitions, the total number of carbon atoms on a substituent is indicated by the C _ij prefix, where the numbers i and j define the number of carbon atoms; this definition includes straight-chain alkyl, branched-chain alkyl, and cycloalkyl or (cycloalkyl)alkane base group.

It is important to realize that when substituents are introduced into a given structural unit, they may be present singly or in plural. For example, halogen (referring to fluorine, chlorine, bromine or iodine) substitution may indicate that the unit to which it is attached may be substituted by one or more halogen atoms, which may be the same or different.

synthesis

Compounds of formula I can be prepared by one of several synthetic methods. For example, appropriately substituted indazole-3-carbaldehyde (1), which can be prepared from the corresponding indazole by known methods [J. Med. Chem., 38, 2331 (1995)] , condensation with a suitable nitroalkane to give the corresponding nitroalkene (2), _the resulting product can be reduced with, for example, LiAlH and, if desired, dealkylated with, for example, boron tribromide to give the desired compound of formula I 3.

Reaction scheme 1

Another method for preparing compounds of formula I is outlined in Scheme 2. Suitably substituted 2-fluoro-acetophenones (4) were purchased or prepared by known methods such as Tetrahedron, 50, 1179 (1994) in the presence of a strong base such as lithium diisopropylamide, with the desired Aldehydes (such as acetaldehyde) react to produce aryl β-hydroxyalkyl ketones (5) [Synthetic Communication (Synth.Commun.), 10, 851 (1980)], and the resulting product is substituted with anhydrous hydrazine by known methods Indazole 6 [Journal of Medical Chemistry, 37, 2721 (1994)]. The secondary alcohol moiety of 6 is converted to the desired primary amine by a well-known sequence, including initial activation by formation of a sulfonated ester, followed by displacement of the ester by reaction with sodium azide, and finally simultaneous reduction of the azide to remove any phenol A protecting group (such as benzyl) to yield the desired amine (3). Alternatively, reaction of compound 4 with an aldehyde such as acetaldehyde under acidic conditions yields a chalcone intermediate such as 7 [J. Chem. Soc., 2403 (1953)] (Scheme 3). Addition of a suitable protective amine, such as benzylamine, to the double bond of 7 yields the desired aminoketone 8 [Chem.Pharm.Bull. 22, 1348 (1974)] when given as Hydrazine treatment of 8 yielded substituted indazole 9 [J. Med. Chem., 37, 2721 (1994)]; removal of the protective group by hydrogenolysis yielded the desired compound 3.

Reaction flow 2

Reaction scheme 3

Another method for the preparation of compounds of structural formula I is also described in Scheme 4 from (5-benzyloxy-1H-indazol-3-yl)-acetic acid (10) [Journal of the American Chemical Society, 79, 5246 (1959)] started. Reaction of 10 with acetic anhydride under Dakin-West conditions in the presence of an appropriate base [Chem. -Hydroxylamine reaction produces oxime 12. Reduction of 12 with eg borane will yield the desired compound 13 of formula I [Eur. J. Med. Chem., 27, 595 (1992), Tetrahedron, 29, 223 (1988)].

Reaction scheme 4

Compounds of formula I where R is C(=O)W can be prepared by combining an appropriate indazole 3, or preferably an appropriate amino-protected intermediate such as 14 (Scheme 5) with the desired activated Acid derivatives (such as acid halides or activated esters, etc.) react to produce ester 15. Removal of the N-protecting group from intermediate 15 yields the desired compound 16 of structural formula I.

Reaction scheme 5

The compounds of formula I of the present invention may be incorporated into various types of ophthalmic formulations for delivery to the eye (eg, topically, intrathecally or via implants). The compounds are preferably incorporated into topical ophthalmic formulations for delivery to the eye. Sterile aqueous ophthalmic suspensions or solutions can be prepared by combining these compounds with ophthalmically acceptable preservatives, surfactants, viscosity builders, penetration enhancers, buffers, sodium chloride and water. Ophthalmic solution formulations can be prepared by dissolving the compound in a physiologically acceptable isotonic aqueous buffer. Additionally, the ophthalmic solutions may include an ophthalmically acceptable surfactant to aid in solubilization of the compound. In addition, the ophthalmic solution may contain viscosity-increasing agents, such as hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, to improve retention of the formulation in the conjunctival sac. element, methylcellulose, polyvinylpyrrolidone, etc. Gelling agents including, but not limited to, gellan gum and xanthan gum may also be used. For the preparation of sterile ophthalmic ointments, the active ingredient is mixed with a preservative in a suitable carrier such as mineral oil, liquid lanolin, or white petrolatum. Sterile ophthalmic gels can be prepared by suspending the active ingredient in a hydrophilic matrix prepared by mixing e.g. Mix in preservatives and tonicity agents.

The compounds are preferably formulated as topical ophthalmic suspensions or solutions at a pH of about 5-8. The compound is generally contained in these preparations in an amount of 0.01% to 5% by weight, and preferably in an amount of 0.25% to 2% by weight. Thus, for topical formulations, 1-2 drops of these formulations are applied to the ocular surface 1-4 times daily, according to the judgment of the experienced clinician.

These compounds can also be used in combination with other agents used in the treatment of glaucoma, such as, but not limited to, beta-blockers (e.g., timolol, betaxolol, levobetaxolol, Lol, levobunolol, propranolol), carbonic anhydrase inhibitors (such as brinzolamide and dorzolamide), alpha ₁ antagonists (such as nipredilol), alpha ₂ agonists (such as iopidine and bromine monidine), miotics (such as pilocarpine and epinephrine), prostaglandin analogs (such as latanoprost, travaprost, unoprostone, and compounds listed in U.S. Pat. ), "low-pressure lipids" (such as lumigan and compounds listed in 5,352,708), and neuroprotective agents (such as compounds from US Patent No. Eleprodil and suitable compounds from WO94/13275, including memantine).

Preferred compounds of formula I are described in Examples 1 and 2. Most preferred is the compound of Example 1. Examples of formulations contemplated suitable for delivering this compound to the eye are provided.

Example 1

3-(2-aminopropyl)-1H-indazol-5-ol hydrochloride

Step A: 1-[5-Benzyloxy-3-(2-oxo-propyl)-indazol-1-yl]-ethanone

A mixture of (5-benzyloxy-1H-indazol3-yl)-acetic acid (2g, 7.08mmol) and sodium acetate (0.99g, 12mmol) in acetic anhydride (6ml) was stirred at 130°C for 3 hours. After cooling, water (15 ml) and ethyl acetate (15 ml) were added to the reaction mixture. The aqueous phase was separated and extracted with ethyl acetate (2 x 15ml). The combined extracts were washed with saturated aqueous NaHCO ₃ (2×20 ml) and saturated aqueous NaCl (20 ml), dried (MgSO ₄ ) and evaporated to a residue which was chromatographed (silica, 15% ethyl acetate in hexanes) to obtain a yellow solid (0.48 g): ¹ H NMR (CD ₃ OD) δ 8.35-8.30 (m, 1H), 7.48-7.23 (m, 6H), 7.04-7.02 ( m, 2H), 5.11(s, 2H), 4.02(s, 2H), 2.77(s, 3H); ¹³ C NMR (CDCl ₃ ) δ203.64(C), 170.47(C), 156.20(C), 145.04(C), 136.52(C), 128.64(CH), 128.31(CH), 127.60(CH), 120.51(CH), 116.74(CH), 102.26(CH), 70.68(CH ₂ ), 42.76(CH ₂ ), 29.59 (CH ₃ ), 22.77 (CH ₃ ), MS (APCl) m/z 323 (M+H) ⁺ .

Step B: 1-[5-Benzyloxy-3-(2-hydroxypropyl)-indazol-1-yl]-ethanone

To a solution of the product from Step A (0.13g, 0.4mmol) in methanol (8ml) was added _NaBH4 (0.016g, 0.4mmol) and the mixture was stirred at room temperature for 20 minutes. Then saturated aqueous _NH4Cl (20ml) and ethyl acetate (20ml) were added to the reaction mixture. The aqueous phase was separated and extracted with ethyl acetate (3 x 15ml). The combined extracts were washed with saturated aqueous NaCl (2 x 15ml), dried ( _MgSO4 ) and evaporated to an oil (0.12g). MS (APCl) m/z 325 (M+H) ⁺ .

Step C: 1-[3-(2-Azido-propyl)-5-benzyloxy-indazol-1-yl]-ethanone

A solution of the product from Step B (0.12g, 0.37mmol) and methanesulfonyl chloride (0.04ml, 0.48mmol) in _CH2Cl2 ( _5ml ) was cooled to 0°C in the presence of nitrogen, and triethylamine (0.07ml , 0.48mmol). The resulting mixture was stirred at 0°C for 10 minutes, then saturated aqueous _NH4Cl (20ml) and ethyl acetate (20ml) were added. The aqueous phase was separated and extracted with ethyl acetate (2 x 5ml). The combined extracts were washed with saturated aqueous NaCl (2 x 15ml), dried ( _MgSO4 ) and evaporated to a residue which was dissolved in DMF (3ml). Sodium azide (0.08 g, 1.2 mmol) was added, and the mixture was stirred at 70°C for 18 hours. After cooling, water (20ml) and diethyl ether (20ml) were added, the aqueous phase was separated and further extracted with diethyl ether (3 x 20ml). The combined extracts were washed with saturated aqueous NaCl (3 x 15 ml), dried ( _MgSO4 ) and evaporated to a residue which was chromatographed (silica, 10% ethyl acetate in hexanes) Purification afforded a yellow oil (0.12 g). MS (ES) m/z 350 (M+H) ⁺ .

Step D: 3-(2-Aminopropyl)-1H-indazol-5-ol hydrochloride

A solution of the product from Step C (0.12 g, 0.34 mmol) in methanol (20 mL) was shaken under hydrogen atmosphere (35 psi) in the presence of 10% palladium on carbon (0.12 g) for 18 hours. The catalyst was removed by filtration and the filtrate was evaporated to a residue which was purified by chromatography to give an oil. Treatment of the oil with 1 N HCl in ethanol afforded the hydrochloride salt (0.013 g) as a colorless semi-solid: ¹ H NMR (CD ₃ OD) δ 7.34-7.30 (m, 1H), 7.05-6.98 (m , 2H), 7.04-7.02(m, 2H), 3.68-3.58(m, 1H), 3.19-3.05(m, 2H), 1.19-1.16(d, J=6Hz, 3H), MS(ES) m/ z 192 (M+H) ⁺ . Treatment of the oil resulting from this step with fumaric acid afforded the fumarate salt as a gray solid; mp 227-230°C. Analysis: Calculated for _{C10H13N3O - C4H4O4-0.3H2O} : C, _53.77 ; H, _5.63 ; _N , _13.43 . Results: C, 53.83; H, 5.85; N, 13.34.

Example 2

3-(2-Aminopropyl)-1-methyl-1H-indazol-5-ol fumarate

Step A. 1-(5-Benzyloxy-1H-indazol-3-yl)-propan-2-one

A solution of the product from Example Step 1 (2.0 g, 6.2 mmol) and sodium hydroxide (0.3 g, 7.5 mmol) in a mixture of methanol (15 mL) and water (15 mL) was stirred at room temperature for 18 hours. The reaction mixture was extracted with ethyl acetate (4 x 30ml), the combined extracts were washed with brine, dried ( _MgSO4 ) and evaporated to a residue which was chromatographed (silica, ethyl acetate/ Hexane, 1:1) to obtain a syrup (1.5 g): ESI/MS m/z 281 (M+H) ⁺ .

Step B. 1-(5-Benzyloxy-1-methyl-1H-indazol-3-yl)-propan-2-one

To a solution of the product from Step A (1.2g, 4.28mmol) in DMF (10ml) was added iodomethane (0.53ml, 8.6mmol) and potassium carbonate (1.2g, 8.6mmol); the mixture was stirred at 70°C 16 hours. After adding water (15ml) and ethyl acetate (15ml) to the reaction mixture, the aqueous phase was separated and extracted with ethyl acetate (3 x 20ml). The combined extracts were washed with brine, dried ( _MgSO4 ) and evaporated to a residue which was purified by chromatography (silica, ethyl acetate/hexanes, 1:1) to give a viscous oil (0.6 g): APCl/LCMS m/z 295 (M+H) ⁺ .

Step C. 3-(2-Aminopropyl)-1-methyl-1H-indazol-5-ol fumarate

By treating the product of step B (0.3 g, 0.1 mmol) in a similar manner as described in step B to step D of Example 1, an oil was obtained which was converted to the fumarate salt (0.071 g): mp 136-139°C ; LCMS m/z 206 (M+H) ⁺ . Analysis: Calculated for _C11H15N3O ·C4H4O4 _{· 0.1H2O} : _{C , 55.76} ; _{H , 5.98} ; _N , 13.00. Results: _C , 55.53; _H , 6.11; _N , 13.22.

Example 3

  2-Methyl-propionic acid 3-(2-aminopropyl)-1H-indazol-5-yl ester

Step A: 3-(2-(9-Fluorenylmethoxycarbonamido)propyl)-1H-indazol-5-ol

To a mixture of dioxane and water (4:1, 10 mL) was added 3-(2-aminopropyl)-1H-indazol-5-ol (0.10 g, 0.36 mmol), 9-fluorenylmethoxy Carbonyl chloride (0.13 g, 0.54 mmol) and sodium bicarbonate (0.9 g, 0.54 mmol). The reaction mixture was poured into dilute sodium bicarbonate, and the resulting mixture was extracted with ether. The combined organic extracts were dried ( _MgSO4 ) and concentrated, and the resulting residue was purified by chromatography.

Step B: 2-Methyl-propionic acid 3-(2-(9-fluorenylmethoxycarbonamido)propyl)-1H-indazol-5-yl ester

To 3-(2-(9-fluorenylmethoxycarbonamido)propyl)-1H-indazol-5-ol (0.17g, 0.41mmol) and diisopropylethylamine ( 0.06 g, 0.50 mmol) in dichloromethane (10 mL) was added 2-methylpropionyl chloride (0.05 g, 0.5 mmol) followed by 4-dimethylaminopyridine (0.05 g, 0.4 mmol). The reaction can be warmed to room temperature and stirred at room temperature. The reaction mixture was added to dilute aqueous sodium bicarbonate and extracted with ether. The ether extracts were combined, washed, dried ( _MgSO4 ) and concentrated. The residue was purified by chromatography.

Step C: 2-Methyl-propionic acid 3-(2-aminopropyl)-1H-indazol-5-yl ester

2-Methyl-propionic acid 3-(2-(9-fluorenylmethoxycarbonamido)propyl)-1H-indazol-5-yl ester (0.16g, 0.33mmol) in piperidine and di A solution in a 1:4 mixture of methylformamide (2.5 mL) was stirred at ambient temperature. The reaction mixture was poured into dilute aqueous sodium bicarbonate, and the resulting mixture was extracted first with ethyl acetate and then with dichloromethane. The combined organic extracts were dried ( _MgSO4 ) and concentrated. The residue was purified by chromatography.

Receptor and binding agonist activity according to the invention can be determined using the methods described below.

method 1

5- _HT2 receptor binding test

To determine the affinity of serotonergic compounds for the 5-HT ₂ receptor, their ability to compete with the agonist radioligand [ ¹²⁵ I]DOI for binding to the brain 5-HT ₂ receptor was as follows with a slight modification of the literature method [Neuropharmacology (Neuropharmacology), 26, 1803 (1987)]. Aliquots (400 μl) of postmortem rat or human cerebral cortex homogenates dispersed in 50 mM TrisHCl buffer (pH 7.4) in the presence or absence of methiothepin (final concentration 10 μM) were mixed with [ ¹²⁵ I]DOI (final concentration 80 pM) were incubated to determine total and non-specific binding, respectively, in a total reaction volume of 0.5 ml. Test mixtures were incubated in polypropylene tubes for 1 hour at 23°C and terminated by rapid vacuum filtration through WhatmanGF/B glass fiber filters pre-soaked in 0.3% polyethyleneimine in ice-cold buffer middle. Methionine was replaced with test compound (at different concentrations). Filter-bound radioactivity was determined by scintillation spectrometry on a beta-counter. Data were analyzed using a non-linear iterative curve fitting computer program [Trends Pharmacol. Sci., 16, 413 (1995)] to determine the affinity parameters of the compounds. The maximum compound concentration required to inhibit 50% of [ ¹²⁵ I]DOI binding was defined as the IC ₅₀ or K _i value.

Method 2

5-HT ₂ Functional Test: Phosphoinositide (PI) Conversion Test

The ability of compounds to stimulate [ ³ H]inositol phosphate production (via their ability to activate phospholipase C) in [ ³ H]inositol-labeled A7r5 rat vascular smooth muscle cells can be assayed in vitro for the effect of serotonergic compounds on Relative agonist activity of the 5- _HT2 receptor. These cells were grown on the culture plate, and the culture plate was maintained in a humid environment of 5% CO ₂ and 95% air, and supplemented with 4.5 g/l glucose and supplemented with 2 mM glutamine, 10 μg/ml genta Dulbecco's Modified Eagle's Medium (DMEM) with Mycin and 10% Fetal Bovine Serum. In order to achieve the purpose of carrying out the phosphoinositide (PI) conversion experiment, as previously described [Journal of Pharmacology and Experimental Therapeutics (J.Pharmacol.Expt.Ther.), 286, 411 (1998)] cultured A7r5 cells. Confluent cells were exposed to 1.5 μCi [ ³ H]-inositol (18.3 Ci/mmol) in 0.5 ml serum-free medium for 24-30 hours. The cells were then washed once with DMEM/F-12 containing 10 mM LiCl, followed by incubation with the test agent (or solvent as a control) in 1.0 ml of the same medium at 37° C. for 1 hour, after which the medium was aspirated, and Add 1 ml of cold 0.1M formic acid to stop the reaction. [ ³ H]-inositol phosphates ([ ³ H]-IPs) were chromatographically separated on an AG-1-X8 column as previously described [Journal of Pharmacology and Experimental Therapeutics 286, 411 (1998)], wherein Washing sequentially with H ₂ O and 50 mM ammonium formate followed by elution of the total [ ³ H]-IPs fraction with 1.2 M ammonium formate containing 0.1 M formic acid. The eluate (4 ml) was collected, mixed with 15 ml of scintillation fluid and total [ ³ H]-IPs were determined by scintillation counting on a β-counter. Concentration-response data were analyzed by the sigmoid curve fitting function of Origin Scientific Graphics software (Microcal Software, Northampton, MA) to determine agonist potency ( _EC50 values) and efficacy ( _Emax ). Serotonin (5-HT) was used as a positive control (standard) agonist compound, and the potency of the test compound was compared to that of 5-HT (set as 100%). The maximum compound concentration required to stimulate 50% of the production of [ ³ H]-IPs was referred to as the EC ₅₀ value.

The method described above was used to generate the data shown in Table 1.

Table 1. 5-HT ₂ Receptor Binding and Functional Data compound IC50, nM EC50, nM Efficacy (E _max %) 3-(2-aminopropyl)-1H-indazol-5-ol hydrochloride 2.5 1,210 97 3-(2-Aminopropyl)-1-methyl-1H-indazol-5-ol fumarate - 778 98

Example 4 Element Content (wt%) 3-(2-Aminopropyl)-1-methyl-1H-indazol-5-ol fumarate 0.01-2% Hydroxypropylmethylcellulose 0.5% Disodium Hydrogen Phosphate (Anhydrous) 0.2% Sodium chloride 0.5% Disodium EDTA (Disodium Ethylenediaminetetraacetic Acid) 0.01% Polysorbate 80 0.05% Germine 0.01% Sodium hydroxide/hydrochloric acid for pH adjustment purified water Sufficient to 100%

Example 5 Element Content (wt%) 3-(2-Aminopropyl)-1H-indazol-5-ol 0.01-2% Methylcellulose 4.0% Disodium Hydrogen Phosphate (Anhydrous) 0.2% Sodium chloride 0.5% Disodium EDTA (Disodium Ethylenediaminetetraacetic Acid) 0.01% Polysorbate 80 0.05% Germine 0.01% Sodium hydroxide/hydrochloric acid for pH adjustment pure water Sufficient to 100%

Example 6 Element Content (wt%) 3-(2-Aminopropyl)-1H-indazol-5-ol 0.01-2% guar gum 0.4-6.0% Disodium Hydrogen Phosphate (Anhydrous) 0.2% Sodium chloride 0.5% Disodium EDTA (Disodium Ethylenediaminetetraacetic Acid) 0.01% Polysorbate 80 0.05% Germine 0.01% Sodium hydroxide/hydrochloric acid for pH adjustment pure water Sufficient to 100%

Example 7 Element Content (wt%) 2-Methyl-propionic acid-3-(2-aminopropyl)-1H-indazol-5-phenol ester 0.01-2% White petrolatum and mineral oil and lanolin ointment consistency Disodium Hydrogen Phosphate (Anhydrous) 0.2% Sodium chloride 0.5% Disodium EDTA (Disodium Ethylenediaminetetraacetic Acid) 0.01% Polysorbate 80 0.05% Germine 0.01% Sodium hydroxide/hydrochloric acid for pH adjustment

Claims (9)

1. be used to reduce and control the method for normal or the intraocular pressure that raise, this method comprises compound and the pharmaceutically useful salt of this compounds and the solvate of the following structural formula of using pharmacy effective dose: structural formula I

Wherein G is selected from hydrogen, halogen or C _1-4Alkyl;

R is hydrogen, C _1-4Alkyl, C (=O) W or P (=O) (OX) (OY),

R ¹And R ²Be hydrogen;

R ³And R ⁴Be independently selected from hydrogen, C _1-4Alkyl, or R ³, R ⁴Form cyclopropyl rings with the carbon atom that they connected, or in addition, R ²And R ³Common formation (CH ₂) _mTo form saturated heterocycle;

Work as R ²And R ³During for a heterocyclic part, R ¹Can be hydrogen or C _1-4Alkyl;

R ⁵Can be hydrogen or C _1-4Alkyl;

As R, R ¹, R ², R ⁵When all being hydrogen with G, R ³, R ⁴Can not be hydrogen simultaneously;

W is C _1-6Alkyl, NR ⁶R ⁷, N (R ⁶) CH ₂(CH ₂) _nC (=O) N (R ⁷) (R ⁸), OC _1-6Alkyl, C _1-6(it can be by halogen, hydroxyl, CO for alkyl ₂C _1-4Alkyl, CON (C _1-4Alkyl) ₂, C (=NH) NH ₂, HC (=NH) NH ₂, NH ₂Replacement), C _2-4Alkenyl (replaced by phenyl, not by or by C _1-4Alkyl, C _1-4One or more replacements in the alkoxy or halogen);

R ⁶, R ⁷And R ⁸Be independently selected from hydrogen or C _1-4Alkyl;

X and Y are independently selected from hydrogen, C _1-10Alkyl, or X and Y can form rudimentary (CH jointly ₂) _mAlkyl chain;

M is 2-4;

N is 1 or 2.

2. the process of claim 1 wherein that compound is defined as follows:

G is selected from hydrogen, halogen or C _1-4Alkyl;

R be C (=O) W or P (=O) (OX) (OY),

R ¹And R ²Be hydrogen;

R ³, R ⁴Form cyclopropyl rings with the carbon atom that they connected, or in addition, R ²And R ³Common formation (CH ₂) _mTo form saturated heterocycle;

Work as R ²And R ³During for a heterocyclic part, R ¹Can be hydrogen or C _1-4Alkyl;

R ⁵Can be hydrogen or C _1-4Alkyl;

W is C _1-6Alkyl, NR ⁶R ⁷, N (R ⁶) CH ₂(CH ₂) _nC (=O) N (R ⁷) (R ⁸), OC _1-6Alkyl, C _1-6(it can be by halogen, hydroxyl, CO for alkyl ₂C _1-4Alkyl, CON (C _1-4Alkyl) ₂, C (=NH) NH ₂, HC (=NH) NH ₂, NH ₂Replacement), C _2-4Alkenyl (replaced by phenyl, not by or by C _1-4Alkyl, C _1-4One or more replacements in the alkoxy or halogen);

R ⁶, R ⁷And R ⁸Be independently selected from hydrogen or C _1-4Alkyl;

X and Y are independently selected from hydrogen, C _1-10Alkyl, or X and Y can form rudimentary (CH jointly ₂) _mAlkyl chain;

M is 2-4;

N is 1 or 2.

3. the process of claim 1 wherein that compound is defined as follows:

G is selected from hydrogen, halogen or C _1-4Alkyl;

R be hydrogen, C (=O) W or P (=O) (OX) (OY),

R ¹And R ²Be hydrogen;

R ³And R ⁴Be independently selected from hydrogen, C _1-4Alkyl, or R ³, R ⁴Form cyclopropyl rings with the carbon atom that they connected;

R ⁵Can be hydrogen or C _1-4Alkyl;

W is C _1-6Alkyl, NR ⁶R ⁷, N (R ⁶) CH ₂(CH ₂) _nC (=O) N (R ⁷) (R ⁸), C _1-6(it can be by halogen, hydroxyl, CO for alkyl ₂C _1-4Alkyl, CON (C _1-4Alkyl) ₂, C (=NH) NH ₂, HC (=NH) NH ₂, NH ₂Replacement), C _2-4Alkenyl (replaced by phenyl, not by or by C _1-4Alkyl, C _1-4One or more replacements in the alkoxy or halogen);

R ⁶, R ⁷And R ⁸Be independently selected from hydrogen or C _1-4Alkyl;

X and Y are independently selected from hydrogen, C _1-10Alkyl, or X and Y can form rudimentary (CH jointly ₂) _mAlkyl chain;

M is 2 or 3;

N is 1 or 2.

4. the process of claim 1 wherein that compound is defined as follows:

G is selected from hydrogen, halogen or C _1-4Alkyl;

R is hydrogen or C (=O) W;

R ¹And R ²Be hydrogen;

R ³Be hydrogen, and R ⁴Be methyl, perhaps R ³, R ⁴Form cyclopropyl rings with the carbon atom that they connected;

R ⁵Be hydrogen;

W is C _1-6Alkyl, C _1-6(it can be by halogen, hydroxyl, CON (C for alkyl _1-4Alkyl) ₂, C (=NH) NH ₂Replace).

5. the process of claim 1 wherein that compound is selected from:

3-(2-aminopropyl)-1H-indazole-5-phenol;

3-(2-aminopropyl)-1-methyl isophthalic acid H-indazole-5-phenol;

2-(5-methoxyl group-1H-indazole-3-yl)-1-methyl-ethamine;

3-(2-aminopropyl)-6-fluoro-1H-indazole-5-phenol;

3-(2-aminopropyl)-7-methyl isophthalic acid H-indazole-5-phenol;

3-(2-aminopropyl)-6-fluoro-1-methyl isophthalic acid H-indazole-5-phenol;

2-methyl-propionic acid 3-(2-aminopropyl)-1H-indazole-5-base ester;

2,2-dimethyl-propionic acid 3-(2-aminopropyl)-1H-indazole-5-base ester;

Cyclopropane-carboxylic acid 3-(2-aminopropyl)-1H-indazole-5-base ester;

N, N-diethyl-succinamic acid 3-(2-aminopropyl)-1H-indazole-5-base ester.

6. the compound of following structural formula:

Wherein G is selected from hydrogen, halogen or C _1-4Alkyl;

R be C (=O) W or P (=O) (OX) (OY),

R ¹And R ²Be hydrogen;

R ³, R ⁴Form cyclopropyl rings with the carbon atom that they connected, or in addition, R ²And R ³Common formation (CH ₂) _mTo form saturated heterocycle;

Work as R ²And R ³During for a heterocyclic part, R ¹Can be hydrogen or C _1-4Alkyl;

R ⁵Can be hydrogen or C _1-4Alkyl;

W is C _1-6Alkyl, NR ⁶R ⁷, N (R ⁶) CH ₂(CH ₂) _nC (=O) N (R ⁷) (R ⁸), OC _1-6Alkyl, C _1-6(it can be by halogen, hydroxyl, CO for alkyl ₂C _1-4Alkyl, CON (C _1-4Alkyl) ₂, C (=NH) NH ₂, HC (=NH) NH ₂, NH ₂Replacement), C _2-4Alkenyl (replaced by phenyl, not by or by C _1-4Alkyl, C _1-4One or more replacements in the alkoxy or halogen);

R ⁶, R ⁷And R ⁸Be independently selected from hydrogen or C _1-4Alkyl;

X and Y are independently selected from hydrogen, C _1-10Alkyl, or X and Y can form rudimentary (CH jointly ₂) _mAlkyl chain;

M is 2-4;

N is 1 or 2.

7. the compound of claim 6, wherein:

G is selected from hydrogen, halogen or C _1-4Alkyl;

R be hydrogen, C (=O) W or P (=O) (OX) (OY),

R ¹And R ²Be hydrogen;

R ³And R ⁴Be independently selected from hydrogen, C _1-4Alkyl, or R ³, R ⁴Form cyclopropyl rings with the carbon atom that they connected;

R ⁵Can be hydrogen or C _1-4Alkyl;

W is C _1-6Alkyl, NR ⁶R ⁷, N (R ₆) CH ₂(CH ₂) _nC (=O) N (R ⁷) (R ⁸), C _1-6(it can be by halogen, hydroxyl, CO for alkyl ₂C _1-4Alkyl, CON (C _1-4Alkyl) ₂, C (=NH) NH ₂, HC (=NH) NH ₂, NH ₂Replacement), C _2-4Alkenyl (replaced by phenyl, not by or by C _1-4Alkyl, C _1-4One or more replacements in the alkoxy or halogen);

R ⁶, R ⁷And R ⁸Be independently selected from hydrogen or C _1-4Alkyl;

X and Y are independently selected from hydrogen, C _1-10Alkyl, or X and Y can form rudimentary (CH jointly ₂) _mAlkyl chain;

M is 2 or 3;

N is 1 or 2.

8. the compound of claim 6, wherein:

G is selected from hydrogen, halogen or C _1-4Alkyl;

R is hydrogen or C (=O) W;

R ¹And R ²Be hydrogen;

R ³Be hydrogen, and R ⁴Be methyl, perhaps R ³, R ⁴Form cyclopropyl rings with the carbon atom that they connected;

R ⁵Be hydrogen;

W is C _1-6Alkyl, C _1-6(it can be by halogen, hydroxyl, CON (C for alkyl _1-4Alkyl) ₂, C (=NH) NH ₂Replace).

9. the compound of claim 6, wherein compound is selected from:

3-(2-aminopropyl)-1H-indazole-5-phenol;

3-(2-aminopropyl)-1-methyl isophthalic acid H-indazole-5-phenol;

2-(5-methoxyl group-1H-indazole-3-yl)-1-methyl-ethamine;

3-(2-aminopropyl)-6-fluoro-1H-indazole-5-phenol;

3-(2-aminopropyl)-7-methyl isophthalic acid H-indazole-5-phenol;

3-(2-aminopropyl)-6-fluoro-1-methyl isophthalic acid H-indazole-5-phenol;

2-methyl-propionic acid 3-(2-aminopropyl)-1H-indazole-5-base ester;

2,2-dimethyl-propionic acid 3-(2-aminopropyl)-1H-indazole-5-base ester;

Cyclopropane-carboxylic acid 3-(2-aminopropyl)-1H-indazole-5-base ester;

N, N-diethyl-succinamic acid 3-(2-aminopropyl)-1H-indazole-5-base ester.