AU2001219180B2 - 5-hydroxy indazole derivatives for treating glaucoma - Google Patents

Description

WO 01/70701 PCT/USOO/31143 INDAZOLE DERIVATIVES FOR TREATING GLAUCOMA The present invention is directed to substituted 3-(2-aminoethyl)-1H-indazolsome of which are novel, for lowering and controlling normal or elevated intraocular pressure (IOP) and treating glaucoma.

Background of the Invention The disease state referred to as glaucoma is characterized by a permanent loss of visual function due to irreversible damage to the optic nerve. The several morphologically or functionally distinct types of glaucoma are typically characterized by elevated IOP, which is considered to be causally related to the pathological course of the disease. Ocular hypertension is a condition wherein intraocular pressure is elevated but no apparent loss of visual finction has occurred; such patients are is considered to be a high risk for the eventual development of the visual loss associated with glaucoma. Some patients with glaucomatous field loss have relatively low intraocular pressure. These so called normotension or low tension glaucoma patients can also benefit from agents that lower and control IOP. If glaucoma or ocular hypertension is detected early and treated promptly with medications that effectively reduce elevated intraocular pressure, loss of visual function or its progressive deterioration can generally be ameliorated. Drug therapies that have proven to be effective for the reduction of intraocular pressure include both agents that decrease aqueous humor production and agents that increase the outflow facility. Such therapies are in general administered by one of two possible routes, topically (direct application to the eye) or orally.

There are some individuals who do not respond well when treated with certain existing glaucoma therapies. There is, therefore, a need for other topical therapeutic agents that control IOP.

It has been found that serotonergic compounds which possess agonist activity at 5-HT 2 receptors effectively lower and control normal and elevated IOP and are useful for treating glaucoma, see commonly owned co-pending application, PCT/US99/19888. Compounds that act as agonists at 5-HT2 receptors are well known and have shown a variety of utilities, primarily for disorders or conditions associated with the central nervous system (CNS). U.S. Patent 5,494,928 discloses certain 2- (indol-l-yl)-ethylamine derivatives that are 5-HT 2 c agonists for the treatment of obsessive compulsive disorder and other CNS derived personality disorders. U.S.

-1- WO 01/70701 PCT/US00/31143 Patent 5,571,833 discloses tryptamine derivatives that are 5-HT 2 agonists for the treatment of portal hypertension and migraine. U.S. Patent 5,874,477 discloses a method for treating malaria using 5-HT2A/2C agonists. U.S. Patent 5,902,815 discloses the use of 5-HT2A agonists to prevent adverse effects of NMDA receptor hypofunction. W098/31354A2 discloses 5-HT2B agonists for the treatment of depression and other CNS conditions. Agonist response at the 5-HT2A receptor is reported to be the primary activity responsible for hallucinogenic activity, with some lesser involvement of the 5-HT 2 c receptor possible [Psychopharmacology, Vol. 121:357, 1995].

The present invention is directed to substituted 3-(2-aminoethyl)-lH-indazolsome of which are novel. It is believed that these compounds will have a high affinity for and function as agonists at the serotonergic 5-HT 2 receptor, and will thereby be useful for lowering and controlling normal or elevated intraocular pressure (IOP) and treating glaucoma. When a phenolic moiety is included in this substitution, e.g. a hydroxyl group at indazole ring position five, such compounds can be particularly sensitive to oxidation reactions well known to occur with phenolic compounds in general, including the related hydroxytryptamines Phys. Chem. 103, 8606 (1999), Chem. Res. Toxicol. 11, 639 (1998), J. Org. Chem. 52, 2817 (1987), J.

Pharm. Sci. 77, 911 (1988)], which are of particular relevance to the present application. Protection of such hydroxy substituted phenols from oxidation can be accomplished by derivatization of the aryl hydroxy group to provide a suitable ester, carbamate, or carbonate. Though the ester, carbamate, or carbonate derivatives do not themselves possess a high affinity for the above mentioned receptors, they do have utility in the treatment of glaucoma since suitably protected phenols can be cleaved in vivo either by chemical hydrolysis or through the action of tissue esterases. Such cleavage would deliver the desired therapeutic agent, that is, the desired novel hydroxy-indazole compounds of the present invention. The concept of utilizing 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 with no comment on the utility of the compound Amer. Chem. Soc.

79, 5242 (1957); J. Amer. Chem. Soc. 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 No. W098/30548 (1998). The utility cited for the compounds of this application is -2for treating central nervous system diseases such as sexual disorders, genital C insufficiency, appetite regulation disorders, anxiety, depression, and sleep disorders.

There is not contemplation of ophthalmic use noted in this application. Published SInternational Patent Application No. WO00/12482 (2000) discloses certain l-(indazol- 3-yl)-2-propylamine derivatives that are 5-HT 2 agonists for the treatment of disorders of the central nervous system.

O Throughout the description and claims of this specification, use of the word "comprise" and variations of the word, such as "comprising" and "comprises", is not intended to exclude other additives, components, integers or steps.

The discussion of documents, acts, materials, devices, articles and the like is included in this specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any or all of these matters formed part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed in Australia before the priority date of each claim of this application.

W:\BreeAmendments\76551 Alcon.doc 0 SSummary of the Invention oThe present invention is directed to derivatives of 3-(2-aminoethyl)-lH- 00 some of which are novel, which can be used to lower and control IOP associated with normal-tension glaucoma, ocular hypertension, and glaucoma in warm Sblooded animals, including man. The compounds are formulated in pharmaceutical compositions suitable for topical delivery to the eye.

C N 1 Description of the Preferred Embodiments Compounds that are useful according to the present invention are represented by the following Formula 1.

FORMULA I

R

N

G N R wherein G is chosen from hydrogen, halogen, or C 1 -4alkyl; R is hydrogen, C-4alkyl, or

R

1 and R 2 are hydrogen;

R

3 and R 4 are independently chosen from hydrogen, Ci4alkyl or R, R 4 and the carbon atom to which they are attached can form a cyclopropyl ring, or furthermore, R 2 and

R

3 together can be (CH 2 )m to form a saturated heterocycle; when R 2 and R 3 are part of a heterocycle, R' can be hydrogen or C 1 4alkyl;

R

5 can be hydrogen or C 1 4alkyl; when R, R 2 k 5 and G all are hydrogen R 3 and R 4 cannot both be hydrogen; WO 01/70701 PCT/US00/31143 W is Cl-6alkyl, NR 6

R

7

N(R

6

)CH

2

(CH

2

N(R

7 OCl_-alkyl, Cl-6alkyl (which can be substituted with halogen, hydroxyl, CO 2 Ci-4alkyl, CON(Ci-4alkyl)2, NH)N C(=NH)NH 2

NH

2 C2 4 alkcnyl (substituted by phenyl, unsubstituted or substituted with one or more of C.

4 alkyl, Ci4alkoxy or halogen); s R 6

R

7

R

8 are independently chosen from hydrogen or C 1 4alkyl; X and Y are independently chosen from hydrogen, C 1 lo alkyl or X and Y can together form a lower alkyl chain of (CH 2 )m; m is 2 4; n is 1 or 2; 1o and pharmaceutically acceptable salts and solvates of the compounds of Formula I.

Compounds that are novel and which are useful according to the present invention can be defined as follows: is G is chosen from hydrogen, halogen, or C 1 -4alkyl; R is or

R

1 and R 2 are hydrogen;

R

3

R

4 and the carbon atom to which they are attached can form a cyclopropyl ring, or furthermore, R 2 and R 3 together can be (CH2)m to form a saturated heterocycle; when R 2 and R 3 are part of a heterocycle, R' can be hydrogen or C 1 -4alkyl;

R

5 can be hydrogen or Cl-4alkyl; W is Ci- 6 alkyl, NR 6

R

7

N(R

6

)CH

2

(CH

2 OC1.6alkyl, Cl 6 alkyl (which can be substituted with halogen, hydroxyl, CO 2 Ci- 4 alkyl, CON(C-4alkyl) 2

C(=NH)NH

2 HC(=NH)NH2, NH 2

C

2 4 alkenyl (substituted by phenyl, unsubstituted or substituted with one or more of C _4alkyl, Ci4alkoxy or halogen);

R

6

R

7 R are independently chosen from hydrogen or Ci 4 alkyl; X and Y are independently chosen from hydrogen, C 1 lo alkyl or X and Y can together form a lower alkyl chain of(CH 2 )m; mis 2 4; n is 1 or 2.

Preferred compounds are: G is chosen from hydrogen, halogen, or C1.4alkyl; R is hydrogen, or R' and R 2 are hydrogen;

R

3 and R 4 are independently chosen from hydrogen, C 1 4alkyl or R 3

R

4 and the carbon atom to which they are attached can form a cyclopropyl ring;

R

5 can be hydrogen or C 1 _4alkyl; WO 01/70701 WO 0170701PCT[USOO/31 143 W is C 16 alkyl, NR 6

N(R

6

)CH

2

(CH

2 N(R 7

)(R

8 C,-,alkyl (which can be substituted with halogen, hydroxyl, CO 2 CI-4alkyl, CON(Ci 4 alkyl) 2

C(-NH)NH

2

HC(=NH)NE-

2

NH

2

C

24 alkenyl (substituted by phenyl, unsubstituted or substituted with one or more of C I alkyl, C 14 alkoxy or halogen); R67 R, R7, R8are independently chosen from hydrogen or C 1 4 alkyl; X and Y are independently chosen from hydrogen, C, 1 o alkyl or X and Y can together form a lower alkyl chain Of (CH 2 )m; m is 2 or 3; n is 1 or 2.

Most preferred compounds are: is chosen from hydrogen, halogen, or Cl-4alkcyl; R is hydrogen or C(=O)W; *RI and R 2 are hydrogen; is R 3 is hydrogen and R 4 is methyl or R 3

R

4 and the carbon atom to which they are attached form a cyclopropyl ring; R 5 is hydrogen; W is CI- 6 alkyl, C 1 6 alkyl (which can be substituted with halogen, hydroxyl,

CON(C

1 4 alkyl) 2

C(=NIT)NH

2 Representative examples of preferred novel compounds of Formula I are: 3-(2-Aminopropyl)- 3-(2-Aminopropyl)- 1H-indazol-3-yl)-l1-methyl-ethylamine; 3-(2-Aminopropyl)-6-fluoro- 3-(2-Aminopropyl)-7-methyl- 3-(2-Aminopropyl)-6-fluoro-1-methyl- 2-Methyl-propionic acid 3 -(2-aminopropyl)- LH-indazol-5-yl ester;, 2,2-Dimethyl-propionic acid 3 -(2-aminopropyl)- 1H-indazol-5-yI ester; Cyclopropanecarboxylic acid 3-(2-aminopropyl)- IH-indazol-5-yl ester;, N, N-Diethyl-succinamic acid 3-(2-aminopropyl)-1H-indazol-5-yl ester.

It is recognized that compounds of Formula I can contain one or more chiral centers. This invention contemplates all enantiomers, diastercomers and, mixtures thereof.

In the above definitions, the total number of carbon atoms in a substituent group is indicated by the Cj~ prefix where the numbers i and j define the number of WO 01/70701 PCT/US00/31143 carbon atoms; this definition includes straight chain, branched chain, and cyclic alkyl or (cyclic alkyl)alkyl groups.

It is important to recognize that a substituent may be present either singly or S multiply when incorporated into the indicated structural unit. For example, the substituent halogen, which means fluorine, chlorine, bromine, or iodine, would indicate that the unit to which it is attached may be substituted with one or more halogen atoms, which may be the same or different.

SYNTHESIS

The compounds of Formula I can be prepared by using one of several synthetic procedures. For example, condensation of the suitably substituted indazol-3carboxaldehyde which can be prepared from the corresponding indazole by known methods Med. Chem. 38, 2331 (1995)], with the appropriate nitroalkane gives the corresponding nitroalkene which can be reduced with, e.g. LiAlH 4 and if desired dealkylated with, e.g. boron tribromide, to provide the desired compounds 3 of Formula I.

SCHEME 1 CH

CH,

CHO

Y

NO

2 HO NH2 N N N H H

H

1 2 3 Another procedure for preparing compounds of Formula I is outlined in Scheme 2. The appropriately substituted 2-fluoro-acetophenone either purchased or prepared by known procedures, e.g. Tetrahedron 50, 1179 (1994), can be reacted with the desired aldehyde, such as acetaldehyde, in the presence of a strong base, e.g.

lithium diisopropylamide, to give the aryl 1-hydroxyalkyl ketone [Synth.

Commun. 10, 851 (1980)], which can be reacted with anhydrous hydrazine by known methods to provide the substituted indazole 6 Med. Chem. 37, 2721 (1994)].

Conversion of the secondary alcohol moiety of 6 to the desired primary amine can be accomplished using the well known sequence involving initial activation by formation WO 01/70701 PCT/US00/31143 of a sulfonate ester followed by displacement of this ester by reaction with sodium azide, and finally reduction of the azide with concomitant removal of any phenol protective groups, e.g. benzyl, to provide the desired amine Alternately, compound 4 can be reacted with an aldehyde, e.g. acetaldehyde, under acidic conditions to provide the chalcone intermediate, e.g. 7 Chem. Soc., 2403 (1953)] (Scheme Addition of a suitably protected amine, e.g. benzylamine, to the double bond of 7 provides the desired amino ketone 8 [Chem. Pharm. Bull. 22, 1348 (1974)] which when treated with hydrazine provides the substituted indazole 9 Med. Chem.

37, 2721 (1994)]; removal of the protective groups by hydrogenolysis provides the desired compounds 3.

SCHEME 2

CH,

G

CH,

HC-CHO R 1

OH

G

N,H

CH,

HO NH,

N

1. Ms,O 2. NaN, 3. H,

N

CH,

R0O OH SCHEME 3

H,C-CHO

4 H

H-

CH,

R O

G

7 N- Pg CHz RNH, R'

HO

F0

G

8

CH,

HO NH,

H,

CH,

'0N-Pg R

H

-aN G

H

WO 01/70701 PCT/US00/31143 Yet another procedure for preparing compounds of Formula I, but beginning with benzyloxy-1H-indazol-3-yl)-acetic acid (10) Amer. Chem. Soc. 79, 5246 (1959)] as the starting material, is outlined in Scheme 4. Reaction of 10 with acetic anhydride in the presence of the appropriate base under Dakin-West conditions [Chem. Soc.

Rev, 17, 91 (1988)] provides intermediate 11 which upon reaction with O-methylhydroxylamine gives the oxime 12. Reduction of the 12, e.g. with borane, will provide the desired compounds 13 of Formula I [Eur. J. Med. Chem. 27, 595 (1992), Tetrahedron, 29, 223 (1988)].

io SCHEME 4 0 B O H AcO/base 0 0 Bz fv H

H

11 RONH OR

N-OR

Bz.O

CH,

H

12 CH3 HO NH2

H

H,/Pd-C Compounds of Formula I, wherein R is can be prepared by reacting the appropriate indazole 3, or preferably a suitable amino-protected intermediate, e.g.

14 (Scheme 5) with the desired activated acid derivative, such as an acid halide or active ester, or the like, to provide the esters 15. Removal of the N-protective group from the intermediate 15 provides the desired compounds 16 of Formula I.

WO 01/70701 PCT/US00/31143 SCHEME HO NH, HO Protecting HO "k H Group GG

H

3 14

CH

3

CH,

R O NH2 Pr oecting SNH, RpO NN HGroup 0 j N 0 N 16 H G The compounds of this invention, Formula I, can be incorporated into various types of ophthalmic formulations for delivery to the eye topically, intracamerally, or via an implant). The compounds are preferably incorporated into topical ophthalmic formulations for delivery to the eye. The compounds may be combined with ophthalmologically acceptable preservatives, surfactants, viscosity enhancers, penetration enhancers, buffers, sodium chloride, and water to form an aqueous, sterile ophthalmic suspension or solution. Ophthalmic solution formulations may be prepared by dissolving a compound in a physiologically acceptable isotonic aqueous buffer. Further, the ophthalmic solution may include an ophthalmologically acceptable surfactant to assist in dissolving the compound. Furthermore, the ophthalmic solution may contain an agent to increase viscosity, such as, hydroxymethylcellulose, hydroxyethylcellulosc, hydroxypropylmethylcellulose, methylcellulose, polyvinylpyrrolidone, or the like, to improve the retention of the formulation in the conjunctival sac. Gelling agents can also be used, including, but not limited to, gellan and xanthan gum. In order to prepare sterile ophthalmic ointment formulations, the active ingredient is combined with a preservative in an appropriate vehicle, such as, mineral oil, liquid lanolin, or white petrolatum. Sterile ophthalmic gel formulations may be prepared by suspending the active ingredient in a hydrophilic base prepared from the combination of, for example, carbopol-974, or the like, according to the published formulations for analogous ophthalmic preparations; preservatives and tonicity agents can be incorporated.

The compounds are preferably formulated as topical ophthalmic suspensions or solutions, with a pH of about 5 to 8. The compounds will normally be contained in WO 01/70701 PCT/US00/31143 these formulations in an amount 0.01% to 5% by weight, but preferably in an amount of 0.25% to 2% by weight. Thus, for topical presentation 1 to 2 drops of these formulations would be delivered to the surface of the eye 1 to 4 times per day according to the discretion of a skilled clinician.

The compounds can also be used in combination with other agents for treating glaucoma, such as, but not limited to, P-blockers timolol, betaxolol, levobetaxolol, carteolol, levobunolol, propranolol), carbonic anhydrase inhibitors brinzolamide and dorzolamide), al antagonists nipradolol), oa2 agonists iopidine and brimonidine), miotics pilocarpine and epinephrine), prostaglandin analogs latanoprost, travaprost, unoprostone, and compounds set forth in U.S. Patent Nos. 5,889,052; 5,296,504; 5,422,368; and 5,151,444, "hypotensive lipids" lumigan and compounds set forth in 5,352,708), and neuroprotectants compounds from U.S. Patent No. 4,690,931, particularly eliprodil and R-eliprodil, as set forth in a pending application U.S.S.N. 06/203350, and appropriate compounds from W094/13275, including memantine.

The preferred compounds of Formula I are described in Examples 1 and 2.

The most preferred compound is in Example 1. Examples of formulations anticipated to be suitable for delivery of 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-l-yl]-ethanone A mixture of (5-benzyloxy-lH-indazol-3-yl)-acetic acid (2 g, 7.08 mmol) and sodium acetate (0.99 g, 12 mmol) in acetic anhydride (6 ml) was stirred at 130 OC for 3 h. After cooling, water (15 ml) and ethyl acetate (15 ml) were added to the reaction mixture. The aqueous layer was separated and extracted with ethyl acetate (2 x ml). The combined extracts were washed with saturated aqueous NaHCO 3 (2 x ml) and saturated aqueous NaCI (20 ml), dried (MgSO 4 and evaporated to a residue which was purified by chromatography (silica, 15% ethyl acetate in hexane) to give a yellow solid (0.48 'H NMR (CD30D) 8 8.35-8.30 1H), 7.48-7.23 6H), 7.04-7.02 2H), 5.11 2H), 4.02 2H), 2.77 3H), 2.27 3H); 3 C NMR (CDC1 3 6 203.64 170.47 156.20 145.04 136.52 128.64 (CH), 128.31 127.60 120.51 116.74 102.26 70.68 (CH2), 42.76 (CH 2 29.59 (CH 3 22.77 (CH 3 MS(APCI) m/z 323 WO 01/70701 PCT/US00/31143 Step B: 1-[5-Benzyloxy-3-(2-hydroxypropyl)-indazol-1-yl]-ethanone To a solution of the product from Step A (0.13 g, 0.4 mmol) in methanol (8 ml) was added NaBH 4 (0.016 g, 0.4 mmol) and the mixture was stirred at room temperature for 20 min. A saturated aqueous solution of NH 4 Cl (20 ml) and ethyl acetate (20 ml) were added to the reaction mixture. The aqueous layer was separated and extracted with ethyl acetate (3 x 15 ml). The combined extracts were washed with a saturated aqueous solution of NaCI (2 x 15 ml), dried (MgSO 4 and evaporated to give an oil (0.12 MS(APC1) m/z 325 (M+H) Step C: 1-[3-(2-Azido-propyl)-5-benzyloxy-indazol-l-yl]-ethanone A solution of the product from Step B (0.12 g, 0.37 mmol) and methanesulfonyl chloride (0.04 ml, 0.48 mmol) in CH 2 C1 2 (5 ml) and under nitrogen was cooled to 0 °C and triethylamine (0.07 ml, 0.48 mmol) was added. The resulting mixture was stirred at 0 OC for 10 min followed by the addition of a saturated aqueous solution of NH 4 C1 (20 ml) and ethyl acetate (20 ml). The aqucous layer was separated and extracted with ethyl acetate (2 x 5 ml). The combined extracts were washed with a saturated aqueous solution of NaCl (2 x 15 ml), dried (MgSO4), and evaporated to a residue which was dissolved in DMF (3 ml). Sodium azide (0.08 g, 1.2 mmol) was added and the mixture was stirred at 70 OC for 18 h. After cooling, water (20 ml) and ether (20 ml) were added and the aqueous layer was separated and further extracted with ether (3 x 20 ml). The combined extracts were washed with a saturated aqueous solution of NaC1 (3 x 15 ml), dried (MgSO 4 and evaporated to a residue that was purified by chromatography (silica, 10% ethyl acetate in hexane) to give a yellow oil (0.12 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 ml) was shaken under a hydrogen atmosphere (35 psi) in the presence of palladium-on-carbon (0.12 g) for 18h. 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 a 1 N solution of HCI in ethanol gave the hydrochloride salt as a colorless semi-solid (0.013 'H NMR (CD30D) 6 7.34-7.30 1H), 7.05- 6.98 2H), 7.04-7.02 2H), 3.68-3.58 1H), 3.19-3.05 2H), 1.19-1.16 (d, J 6 Hz, 3H), MS(ES) m/z 192 M+H) Treatment of the oil prepared by this 3s procedure with fumaric acid gave the fumarate salt as a gray solid; mp 227-230 0

C.

Analysis: Calculated for C 10 Hi3N 3 0 C 4

H

4 0 4 0.3H20: C, 53.77; H, 5.63; N, 13.43.

Found: C, 53.83; H, 5.85; N, 13.34.

WO 01/70701 PCT/US00/31143 EXAMPLE 2 3-(2-Aminopropyl)-l-methyl-1H-indazol-5-ol fumarate Step A. l-(5-Benzyloxy-1H-indazol-3-yl)-propan-2-one A solution of the product from Step 1 of Example 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 ml) was stirred for 18 h at room temperature. The reaction mixture was extracted with ethyl acetate (4 x 30 ml) and the combined extracts were washed with brine, dried (MgSO 4 and evaporated to a residue which was purified by chromatography (silica, ethyl acetate/hexane, 1:1) to give a syrup (1.5 ESI/MS m/z 281 Step B. 1-(5-Benzyloxy-l-methyl-1H-indazol-3-yl)-propan-2-one To a solution of the product from Step A (1.2 g, 4.28 mmol) in DMF (10 ml) was added iodomethane (0.53 ml, 8.6 mmol) and potassium carbonate (1.2 g, 8.6 mmol); this mixture was stirred for 16 h at 70 0 C. After adding water (15 ml) and ethyl acetate (15 ml) to the reaction mixture, the aqueous layer was separated and extracted with ethyl acetate (3 x 20 ml). The combined extracts were washed with brine, dried (MgSO4), and evaporated to a residue which was puified by chromatography (silica, ethyl acetate/hexane, 1:1) to give a viscous oil (0.6 g): APC/LCMS m/z 295 (M+H) Step C. 3-(2-Aminopropyl)-l-methyl-1H-indazol-5-ol fumarate The product of Step B (0.3 g, 0.1 mmol) was treated in the manner similar to that described in Example 1 Steps B through D to give an oil which was converted to the fumarate salt (0.071 mp 136-139 0 C; LCMS m/z 206 Analysis.

Calculated for ClH 15 sN 3 0 C4H 4 0 4 0.1 H 2 0: C, 55.76; H, 5.98; N, 13.00. Found: C, 55.53; H, 6.11; N, 13.22.

WO 01/70701 PCT/US00/31143 EXAMPLE 3 2-Methyl-propionic acid 3-(2-aminopropyl)-1H-indazol-5-yl ester Step A: 3-(2-(9-Fluorenylmethoxycarbonylamino)propyl)-1H-indazol-5-ol To a mixture ofdioxane and water 10 mL) was added 3-(2-aminopropyl)- (0.10 g, 0.36 mmol), 9-fluorenylmethoxycarbonyl chloride (0.13 g, 0.54 mmol) and sodium bicarbonate (0.9 g, 0.54 mmol). The reaction mixture poured into dilute sodium bicarbonate and the resulting mixture extracted with ether. The combined organic extracts were dried (MgSO 4 and concentrated to a residue that was purified by chromatography.

Step B: 2-Methyl-propionic acid 3-(2-(9 ester To a solution of 3-(2-(9-fluorenylmethoxycarbonylamino)propyl)-lH-indazol- 5-ol (0.17 g, 0.41 mmol) and diisopropylehtylamine (0.06 g, 0.50 mmol), in methylene chloride (10 mL) cooled at 0° C is added 2-methylpropionyl chloride (0.05 g, 0.5 mmol) followed by 4-dimethylaminopyridine (0.05 g, 0.40 mmol). The reaction allowed to warm to room temperature and then is stirred at room temperature. The reaction mixture is added to dilute aqueous sodium bicarbonate and extracted with ether. The combined ether extracts are washed, dried (MgSO4) and concentrated. The residue is purified by chromatography.

Step C: 2-Methyl-propionic acid 3-(2-aminopropyl)-1H-indazol-5-yl ester A solution of 2-methyl-propionic acid fluorenylmethoxycarbonylamino)propyl)-1H-indazol-5-yl ester (0.16g, 0.33 mmol) in a 1: 4 mixture of piperidine and dimethylformamide (2.5 mL) is stirred at ambient temperature. The reaction mixture is poured into dilute aqueous sodium bicarbonate and the resulting mixture is extracted first with ethyl acetate and then with methylene chloride. The combined organic extracts are dried (MgSO 4 and concentrated. The residue is purified by chromatography.

Receptor and binding agonist activity according to this invention can be determined using the following methods.

WO 01/70701 PCT/US00/31143 METHOD 1 2 Receptor Binding Assay To determine the affinities of serotonergic compounds at the 5HT 2 receptors, their ability to compete for the binding of the agonist radioligand [12 I]DOI to brain 2 receptors is determined as described below with minor modification of the literature procedure [Neuropharmacology, 26, 1803 (1987)]. Aliquots of post mortem rat or human cerebral cortex homogenates (400 dispersed in 50 mM TrisHCl buffer (pH 7.4) are incubated with 25 I]DOI (80 pM final) in the absence or presence ofmethiothepin (10 M final) to define total and non-specific binding, respectively, in a total volume of 0.5 ml. The assay mixture is incubated for 1 hour at 23 0 C in polypropylene tubes and the assays terminated by rapid vacuum filtration over Whatman GF/B glass fiber filters previously soaked in 0.3% polyethyleneimine using ice-cold buffer. Test compounds (at different concentrations) are substituted for methiothepin. Filter-bound radioactivity is determined by scintillation spectrometry on a beta counter. The data are analyzed using a non-linear, iterative curve-fitting computer program [Trends Pharmacol. Sci., 16, 413 (1995)] to determine the compound affinity parameter. The concentration of the compound needed to inhibit the 25 I]DOI binding by 50% of the maximum is termed the IC5o or Ki value.

METHOD 2 2 Functional Assay: Phosphoinositide (PI) turnover assay The relative agonist activity of serotonergic compounds at the 5HT 2 receptor can he determined in vitro using the ability of the compounds to stimulate the production of 3 H]inositol phosphates in 3 H]myo-inositol-labeled A7r5 rat vascular smooth muscle cells by their ability to activate the enzyme phospholipase C. These cells are grown in culture plates, maintained in a humidified atmosphere of 5% CO 2 and 95% air and fed semi-weekly with Dulbecco's modified Eagle medium (DMEM) containing 4.5 g/1 glucose and supplemented with 2mM glutamine, 10 jg/ml gentamicin, and 10% fetal bovine serum. For the purpose of conducting the phosphoinositide (PI) turnover experiments, the A7r5 cells are cultured in 24-well plates as previously Pharmacol. Expt. Ther., 286, 411 (1998)]. Confluent cells are exposed for 24-30 hrs to 1.5 tCi 3 H]-myo-inositol (18.3 Ci/mmol) in 0.5 ml of serum-free medium. Cells are then rinsed once with DMEM/F-12 containing 10 mM LiCI prior to incubation with the test agent (or solvent as the control) in 1.0 ml of the same medium for 1 hr at 37'C, after which the medium is aspirated and 1 ml of cold 0.1 M formic acid added to stop the reaction. The chromatographic separation of -14- WO 01/70701 PCT/US00/31143 ['H]-inositol phosphates 3 H]-IPs) on an AG- 1-X8 column is performed as previously described Pharmacol. Expt. Ther. 286, 411 (1998)] with sequential washes with H 2 0 and 50 mM ammonium formatc, 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) is collected, mixed with 15 ml scintillation fluid, and the total 3 H]-IPs determined by scintillation counting on a beta-counter. Concentration-response data are analyzed by the sigmoidal fit function of the Origin Scientific Graphics software (Microcal Software, Northampton, MA) to determine agonist potency (EC5o value) and efficacy (Emax). Serotonin (5HT) is used as a positive control (standard) agonist compound and the efficacy of test compounds is compared to that of 5HT (set at 100%). The concentration of the compound needed to stimulate the production of 3 H]-IPs by 50% of the maximum response is termed the EC 50 value.

The above procedures were used to generate the data shown in Table 1.

TABLE 1 2 Receptor Binding and Functional Data Compound IC 5 o, nM ECs 0 nM Efficacy (Emax, 3-(2-Aminopropyl)-1H-indazol-5-ol 2.5 1,210 97 hydrochloride 3-(2-Aminopropyl)-l -methyl- H- -778 98 fumarate WO 01/70701 WO 0170701PCTUSOO/31 143 EXAMPLE 4 Ingredients J Amount (wt 3-(2-Aminopropy)-l 1-methyl- IH-indazol- 0.01 2%/ fumarate, Hydroxypropyl methylcellulose Dibasic sodium phosphate (anhydrous) 0.2% Sodiu Disodium EDTA (Edetate disodium) 0.0 1% Polyorbae 800.05% Benzlkoium hloide0.01% Sodium hydroxide Hydrochloric acid For adjusting pH Purified water q.s. to 100% EXAMPLE Ingredients Amount (wt 3-(2-Aminopropyl)- lH-indazol-5-ol 0.01 -2% Methyl cellulose Dibasic sodium phosphate (anhydrous) 0.2% Sodium chloride Disodium EDTA (Edetate disodium) 0.01% Polysorbate 80 0.05% Benzalkonium chloride 0.01% Sodium hydroxide Hydrochloric acid For adjusting pH Purified water q.s. to 100% WO 01/70701 WO 0170701PCTIUSOO/31 143 EXAMPLE 6 Ingredients Amount (wt 3-(2-Aminopropyl)- 1H-indazol-5-ol 0.01 -2%4 Guar gumn 0.4- Dibasic sodium phosphate (anhydrous) 0.2%- Sodium chloride Disodium EDTA (Edetate disodium) 0.01% Polysorbate 80 0.05% Benzalkonium chloride 0.01% Sodium hydroxide Hydrochloric acid For adjusting pH Purified water q.s. to 100%/ EXAMPLE 7 Ingredients Amount (wt 2-Methyl-propionic acid 3-(2-aminopropyl)- 0.01 -2% ester White petrolatum and mineral oil and lanolin Ointment consistency Dibasic sodium phosphate (anhydrous) 0.2% Sodium chloride Disodium EDTA (Edetate disodium) 0.01% Polysorbate 80 Benzalkonium chloride Sodium hydroxide Hydrochloric acid 0.05% 0.01% For adjusting pH

Claims (1)

1. 25-03-2002 PCT/USOO/31 143 9. Oktober 2001 Claims 1. Use of a compound of the formula: FORMULA I wherein G is chosen from hydrogen, halogen, or C 1 4alkyl; R is hydrogen, C 1 ~alkyl, O)W, or R' and R 2 are hydrogen; Rand W 4 are independently chosen from hydrogen, C 1 -4alkyl or.R 3 W 4 and the carbon atom to which they are attached can form a cyclopropyl ring, or furthermore, R 2 and R 3 together can be (CH2)M to form a saturated heterocycle; when R 2 and R 3 are part of a heterocycle, R' can be hydrogen or CI-4alkyI; be hydrogen or C1 4 alkyl; when R, R 2 R 5 and G all are hydrogen R 3 and R 4 cannot both be hydrogen; W is C 1 6 alkyl, NRrR 7 N(R6)CH 2 (CH 2 7 )(R 8 0C 18 alkyl, C 1 .6alkyl (substituted with halogen, hydroxyl, CO 2 CI.4alkyl, CON(Cl 1 4alkyl) 2 C(=NH)NH 2 HC(=NH)NH 2 NHA) C 2 4alkenyI (substituted by phenyl, unsubstituted or substituted with one or more of C 14 alkyI, C 14 alkoxy or halogen); R 6 ,R 7 ,R 8 are independently chosen from hydrogen or CI-4alkyl; X and Y are independently chosen from hydrogen, Cl1 alkyl or X and Y can together AMENDED SHEET 25-03-2002 US0031143 form a lower alkyl chain of (CH 2 )m; m is 2 4; n is 1 or 2; and of pharmaceutically acceptable salts and solvates of the compounds for the preparation of a medicament for lowering and controlling normal or elevated intraocular pressure. 2. Use of a compound according to claim 1 wherein in the formula I G is chosen from hydrogen, halogen, or C14alkyl; R is or R' and R 2 are hydrogen; R 3 R 4 and the carbon atom to which they are attached can form a cyclopropyl ring, or furthermore, R 2 and R 3 together can be (CH2)m to form a saturated heterocycle; when R 2 and R 3 are part of a heterocycle, R' can be hydrogen or C 1 .4alkyl; R s can be hydrogen or C 14 alkyl; W is Ci.ealkyl, NR 6 R 7 N(R6)CH 2 (CH 2 OCl.alkyl, C 1 Ialkyl (substituted with halogen, hydroxyl, CO 2 C. 4 alkyl, CON(C.4alkyl) 2 C(=NH)NH HC(NH)NH 2, NH 2 C 2 4alkenyl (substituted by phenyl, unsubstituted or substituted with one or more of C 1 -4alkyl, C 1 .4alkoxy or halogen); R 6 ,R 7 ,R 8 are independently chosen from hydrogen or C 1 -4alkyl; X and Y are independently chosen from hydrogen, C.-10 alkyl, or X and Y can together form a lower alkyl chain of (CH 2 )m; m is 2 4; n is 1 or 2. 3. Use of a compound according to claim 1 wherein in the formula I G is chosen from hydrogen, halogen, or Ci.4alkyl; R is hydrogen, or R' and R 2 are hydrogen; R 3 and R 4 are independently chosen from hydrogen, C 14 alkyl or R 3 R 4 and the carbon atom to which they are attached can form a cyclopropyl ring; R 5 can be hydrogen or Ci4alkyl; W is Cl. 6 alkyl, NR 6 R 7 N(R)CH 2 (CH 2 C 1 .alkyl (substituted with halogen, hydroxyl, CO 2 C-4alkyl, CON(C-4alkyl) 2 C(=NH)NH 2 HC(=NH)NH 2 NH 2 C24alkenyl (substituted by phenyl, unsubstituted or substituted with one or more of C 1 4alkyl, C,. 4 alkoxy or halogen); AMENDED SHEET 25-03-2002 R 6 ,R',R 8 are independently chosen from hydrogen or C 1 -4alkyl; X and Y are independently chosen from hydrogen, C 1 1 0 alkyl or X and Y can together form a lower alkyl chain of (CH 2 )M; m is 2 or 3; n is 1 or 2. 4. Use of a compound according to claim 1 wherein in the formula I G is chosen from hydrogen, halogen, or C 1 4alkyl; R is hydrogen or C(=0)W; R 1 and W 2 are hydrogen; R 3 is hydrogen and W 4 is methyl or R 3 W 4 and the carbon atom to which they are attached form a cyclopropyl ring; R 5 is hydrogen; W is Cl- 6 alkyl or C 1 -6alkyl (substituted with halogen, hydroxyl, CON(C 1 -4alkyl)2, C(=NH)NH 2 Use of a compound according to claim 1 wherein the compound is selected from the group consisting of: 3-(2-aminopropyl)-I 3-(2-aminopropyl)-I -methyl-I 2-(5-methoxy-1 H-indazol-3-yl)-I -methyl-ethylamine; 3-(2..aminopropyl)-6-fluoroI1 3-(2-aminopropyl)-7-nmethyl-I H-indazol-5-01; 3-(2..aminopropyl)-6-fluoro-I -methyl-I H-indazol-5-01; 2-methyl-propionic acid 3-(2..aminopropyl)-I H-indazol-5-yl ester; 2,2-dimethyl-propionic acid 3-(2-aminopropyD)-IH-indazol-5-yI ester; cyclopropanecarboxylic acid 3..(2-aminopropyl)-I H-indazol-5-yl ester; N, N-diethyl-succinamic acid 3-(2-aminopropyl)-1 H-indazol-5-yl ester. US0031143 AMENDED SHEET 25-03-2002 US0031 143 6. A compound of the formula: N R R4 ,N -~N G wherein G is chosen from hydrogen, halogen, or C 1 4alkyl; R is hydrogen, c 1 -4alkyl, or R' and R 2 are hydrogen; R 3 R 4 and the carbon atom to which they are attached can form a cyclopropyl ring, or furthermore, R 2 and W 3 together can be (CHzOM to form a saturated heterocycle and R 4 is hydrogen or C 14 a~kyl; when R 2 and R 3 are part of a heterocycle, R' can be hydrogen or C 1 4 alkyI; can be hydrogen or C 14 alkyl; W is C 1 -6alkyl, NR 6RW, N(R )CH- 2 (CH 2 )IC&O)N(R 7 OC 1 -6alkyl, C 1 .6alkyt (substituted with halogen, hydroxyl, CO 2 C 1 4alkyl, CON(C,4alkyl)2, C(=NH)NH 2 HC(=NH)NH 2 NH- 2 C 2 4alkenyl (substituted by phenyl, unsubstituted or substituted with one or more of C 1 .4alkyl, C 1 -4alkoxy or halogen); R 6 1 RR are independently chosen from hydrogen or C,. 4 alkyl; X and Y are independently chosen from hydrogen, Cl- 1 0 alkyl or X and Y can together form a lower alkyl chain of (CH 2 )m. m is 2 -4; n isi1 or 2. AMENDED SHEET 25-03-2002 7. A compound of the formula R2 N R 3 R0. R 4 R \N G N R s I wherein G is chosen from hydrogen, halogen, or C 14 alkyl; R is or R' and R 2 are hydrogen; R 3 and R 4 are independently chosen from hydrogen, C 1 4alkyl or R 3 R 4 and the carbon atom to which they are attached can form a cyclopropyl ring, R 5 can be hydrogen or C.4alkyl; W is C.-salkyl, NR 6 R 7 N(R6)CH 2 (CH 2 C 1 alkyl (substituted with halogen, hydroxyl, CO 2 C 1 alkyl, CON(CI4alkyl)2, C(=NH)NH 2 HC(=NH)NH 2 NH 2 C 2 4alkenyl (substituted by phenyl, unsubstituted or substituted with one or more of Cl4alkyl, Cl4alkoxy or halogen); R',RT,R' are independently chosen from hydrogen or C. 4 alkyl; X and Y are independently chosen from hydrogen, Cj. 0 o alkyl or X and Y can together form a lower alkyl chain of (CH 2 )m; m is 2 or 3; n is 1 or 2. 8. The compound of Claim 7 wherein: G is chosen from hydrogen, halogen, or C 1 4alkyl; R is C(=O)W; R 1 and R 2 are hydrogen; R 3 is hydrogen and R 4 is methyl or R 3 R 4 and the carbon atom to which they are attached form a cyclopropyl ring; u,0031 143 1 AMENDED SHEET v..S0031 143 25-03-2002 R 5 is hydrogen; W is CI-BalkyI, C 14 6alkyl (substituted with halogen, hydroxyl, CON(Cl- 4 alkyl)2, C( NH)NHA) 9. A compound according to claim 6, wherein G is chosen from hydrogen, halogen, or C14alkyl; R is C(=O)W or P(0)(OX)(OY), R' and R 2 are hydrogen; R 3 R 4 and the carbon atom to which they are attached can form a cyclopropyl ring, or furthermore, R 2 and R 3 together can be (CH2)m to form a saturated heterocycle; when R 2 and R 3 are part of a heterocycle, R 1 can be hydrogen or C 14 alkyI; R 5 can be hydrogen or Cl~alkyl; W is C 1 6 alkyl, NR 6 R 7 N(R6)CH 2 (CH 2 )nC(=O)N(R 7 OC 1 -6alkyl, C 1 .6alkyI (substituted with halogen, hydroxyl, CO 2 CI-4alkyl, CON(Cl~alkyl) 2 C(=NH)NH2, HC(=NH)NH 2 NHA) C 2 4alkenyl (substituted by phenyl, unsubstituted or substituted with one or more of C 1 -4alkyl, 0 1 .4alkoxy or halogen); W, R 7 are independently chosen from hydrogen or C 1 -4alkyl; X and Y are independently chosen from hydrogen, CI- 1 0 alkyl or X and Y can together form a lower alkyl chain of (CH 2 )m mn is 2 -4; n is I or 2. A compound selected from the group consisting of: 3-(2-aminopropyl)-I 3-(2-am inopropyl)-1 -methyl-I 3-(2-aminopropyl)-6-fluoro-I 3-(2-aminopropyI)-7-methyl-I 3-(2-aminopropyl)-6-fluioro-I -methyl-I 2-methyl-propionic acid 3-(2-aminopropyl)-1 H-indazol-5-yi ester; 2,2-dimethyl-propioflic acid 3-(2-aminopropyl)-I H-indazol-5-yl ester; cyclopropanecarboxylic acid 3-(2-aminopropyl)-1 H-indazol-5-yI ester; N, N-diethyl-succinamic acid 3-(2-aminopropyl)-I H-indazot-5-yl ester. 11. Use of a compound according to any of claims 1-5 in combination with one or AMENDED SHEET O more agents for treating glaucoma, selected from the group consisting of p- ci blockers, carbonic anhydrase inhibitors, a, antagonists, a 2 agonists, miotics, Sprostaglandin analogs, hypotensive lipids, and neuroprotectants. 12. Use of a compound according to any one of claims 1-5 in combination with one or more of the following: timolol, betaxolol, levobetaxolol, 0carteolol, levobunolol, propranolol, brinzolamide, dorzolamide, nipradolol, iopidine, brimonidine, pilocarpine, epinephrine, latanoprost, travaprost, unoprostone, Slumigan, eliprodil and R-eliprodil. S13. Composition when used for lowering and controlling normal or elevated intraocular pressure comprising a pharmaceutically effective amount of a compound as used in any one of claims 1-5 and a pharmaceutically acceptable carrier or diluent. 14. Composition for lowering and controlling normal or elevated intraocular pressure comprising a pharmaceutically effective amount of a compound as used in any one of claims 1-5 and a opthalmically acceptable carrier or diluent. Composition according to claim 13 or 14 containing additionally one or more other agents for treating glaucoma. 16. Composition according to claim 15 in which the other agent is selected from the group consisting of 1-blockers, carbonic anhydrase inhibitors, al antagonists, a 2 agonists, miotics, prostaglandin analogs, hypotensive lipids, and neuroprotectants. 17. A method of lowering or controlling normal or elevated intraocular pressure which comprises administering to a person in need thereof an effective amount of a composition according to claim 13 or 14. 18. Use according to claim 1 substantially as hereinbefore described, with reference to any of the Examples. W:BreAmendments~e7551 Aon.doc O 19. A compound according to any one of claims 6, 7 or 10 substantially cN as hereinbefore described, with reference to any of the Examples. A composition according to claim 13 or 14 substantially as 5 hereinbefore described, with reference to any of the Examples. S21. A method according to claim 17 substantially as hereinbefore 0o described, with reference to any of the Examples. DATED: 8 March, 2005 SPHILLIPS ORMONDE FITZPATRICK Attorneys for: ALCON, INC W:\Bree\Amendments\676551 Alcon.doc 24A