JP2003505414A - Azetidine compounds in CNA and eye diseases - Google Patents

Abstract

(57) Abstract: A compound of formula (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for neuroprotection in a subject or for the treatment of cerebral ischemia, central nervous system damage or ocular disease. The use of any of the following salts or prodrugs: wherein R ¹ is aryl; R ² is H, alkyl or aryl; and R ³ is hydrogen or alkyl. According to a further aspect of the invention, the method comprises administering an effective dose of a compound of formula (I), or a pharmaceutically acceptable salt thereof or a prodrug thereof, to a subject in need of such treatment. Methods of treating ischemia, central nervous system damage, or ocular disease are provided.

Description

Detailed Description of the Invention

The present invention relates primarily to neuroprotection and treatment of stroke and other cerebrovascular disorders.

Strokes and other acute brain injuries are a major cause of mortality and morbidity in the adult population. Stroke is the third most common cause of death in the major industrialized nations and is the most common cause of permanent disability. Every year, in the West, about one million people suffer from acute attacks. Twenty-five percent to thirty-five percent of these patients die within the first three weeks, and twenty-five percent to fifty percent of survivors generally rely on family or institutional care for their life expectancy.
The incidence of seizures increases with age and approximately doubles in each of the last 10 years, affecting 30% of people over the age of 65.

Statistics of seizures appear in Western countries in annual incidence rates of 0.1-0.2%, and the global seizure market is estimated to be worth $ 3 billion in 1995, and in 2005 It is expected to rise to $ 10 billion. There is an unmet medical need for cell-conservative treatment of stroke.

Effective neuroprotective therapies are currently not useful for cerebrovascular disorders. The only currently approved therapy for the treatment of ischemic stroke is Genetech's thrombolytic recombinant tissue plasminogen activator (Activase®, rtPA).
; Alteplase). Activase is required for the treatment of acute ischemic stroke in adults for improved neurological recovery and reduced incidence of disability. Treatment with Activase is initiated only after the onset of stroke symptoms and within 3 hours after elimination of intracranial hemorrhage by cranial computed tomography (CT) scan or other diagnostic imaging sensitive to the presence of hemorrhage Should be.

The mechanisms underlying irreversible brain injury that subsequently result in ischemia are complex. Many classes of compounds are currently being investigated as treatments for cerebrovascular disorders. Acute intervention with both cytoprotective (neuroprotective) thrombolytics and other thrombolytics is under active investigation.

Cytoprotective and neuroprotective therapies include drugs that act to prevent cell death during ischemia and reperfusion. These agents include calpain inhibitors, voltage-sensitive calcium channel antagonists and voltage-sensitive sodium channel antagonists, receptor-mediated calcium channel antagonists [N-methyl-D-aspartic acid (NMDA) and α-amino-3-hydroxy-5.
-Methyl-4-isoxazolepropionic acid (AMPA) antagonist], glutamate synthesis inhibitor, glutamate release antagonist, γ-aminobenzoic acid (GABA) antagonist, 5-HT (serotonin) receptor agonist, ganglioside, antioxidant , Growth factors, anti-apoptotic agents (an
tiapoptotic agent) and anti-adhesion molecule (Silver, B
． , Weber, J .; Fisher, M .; , Clin. Neuropharm
acol. 1996, 19, 101-128).

Excitotoxicity is a neuronal death after induction of cerebral ischemia.
) Is a major determinant. Repetitive cell excitation
firing), persistent depolarization, and induction of supernormal ion flux across excitable membranes may initiate a lethal cellular compromise via various synergistic mechanisms during hypoxia excitotoxicity. Control of the excitatory state of a neuron depends on the net subtraction of excitatory and inhibitory effects on the neuron.

In general, primary excitatory effects on neurons are mediated by the glutamatergic system, while primary inhibition is frequently determined by GABAergic innervation. This is because GABA is the main endogenous inhibitory amino acid in the mammalian brain. Thus, increasing the inhibitory effect of GABAergic innervation and decreasing the excitatory effect of glutamate reduces the net excitation of neurons. The decrease in excitement
It reduces the consequences of energy deprivation due to hypoxia and promotes the ability of neurons to withstand hypoxic cerebral ischemia.

[0009] The relatively few drugs currently under investigation as neuroprotective agents for the treatment of stroke and other cerebrovascular disorders are modulators of the endogenous inhibitory amino acid, GABA.

One class of molecules with apparently neuroprotective properties is C-like CI-966.
It is an ABA uptake inhibitor and has been shown to be effective in a gerbil ischemia model utilizing 5 minutes of global cerebral ischemia (Phillis, JW.
, Gen. Pharmacol. 1995, 26, 1061-1064).

The benzodiazepine receptor agonist diazepam has been shown to have several neuroprotective properties (Karle, J. Witt, MR, Niel.
sen, M .; , Brain Res. 1997, 765, 21-29).

[0012]   In rabbits with reversible spinal cord ischemia, muscimol, the reference GABA _A Treatment with agonist 5 mg / kg is₅₀The time was significantly extended. here
, P₅₀Is the duration associated with the resulting 50% probability of permanent paraplegia.
Representation (Madden, K.P., Stroke, 1994, 25, 2271-2)
275).

Felbamate (particularly an antiepileptic drug with GABA agonistic properties) is significantly neuroprotective when administered to all areas of the brain in the gerbil model of transient forebrain ischemia before and after ischemia. Provided. Protection was moderate when felbamate was used before ischemia, but very pronounced when felbamate was given 30 minutes after the attack. Structural protection with felbamate was very pronounced when used during the post-ischemic period (Shuaib, A., Waqar, T.,
Ijaz, M .; S. Kanthan, R .; Wishart, T .; , Howle
tt, W. , Brain Res. 1996, 727, 65-70).

Piracetam is a derivative of GABA and was the first commercial nootropic drug. Piracetam is widely evaluated in the treatment of senile cognitive impairment and dyslexia, but piracetam is also evaluated as a treatment for deficiencies associated with acute stroke. Data from many small short-term studies in patients treated within days of stroke suggest that piracetam is more effective than placebo for the treatment of functional deficiency (Noble., S., Benfield). , P., CNS
Drug 1998, 9, 497-511).

Several combination neuroprotective therapies have been studied in rodent ischemia.
Because the excitotoxic effect of glutamate can be almost completely blocked by GABA in cell cultures, tissue sections, and some animal models. On this basis, a combination of muscimol and MK801, an NMDA receptor antagonist was studied and shown to be effective (Lyde.
n, P. D. Lonzo, L .; , Stroke 1994, 25, 189-1.
96) WO-A-99 / 25353 discloses triazolo [4,3-b] as a benzodiazepine / GABA _A modulator for the treatment of psychotic disorders and neurodegeneration.
The use of pyridazine derivatives is disclosed.

WO-A-90 / 09174 discloses the use of the GABA activator clomethiazole (chlormethiazole) in the prevention and / or treatment of neurodegeneration.
Clomethiazole is believed to act through the GABAergic pathway, whereby clomethiazole potentiates the GABA inhibitory effect on the CNS, giving the drug both hypnotic and neuroprotective properties.

The clinical neuroprotective profile of clomethiazole has been reviewed (Muck
le, H.A. , IDrugs 1999, 2, 184-193). Large Stage I
II trials were conducted in which clomethiazole was evaluated for its ability to reduce nerve damage in acute cerebrovascular ischemia. A subgroup of patients who showed a large seizure experienced significant benefits. 41% of these (n = 545) treated patients were functionally independent after 90 days compared to 30% of placebo patients.

Rat ischemia (Snape, MF, Baldwin, HA, Cross)
, A. J. , Green, A .; R. , Neuroscience 1993, 5
3, 837-844) and gerbil ischemia (Cross, AJ, Jon.
es, J. A. Baldwin, H .; A. , Green, A .; R. , Br. J
． Pharmacol. This GA in 1991, 104, 406-411)
The efficacy of BA modulators has been demonstrated. The dose in the later conceptual scheme is 10
It was 0 mg / kg (intraperitoneal).

Azetidine-1-carboxamide and the use of these compounds in the treatment of all forms of anxiety and epilepsy are described in International Patent Application No. PCT / GB99 / 0.
0224, PCT / GB99 / 00219 and PCT / GB99 / 00223
It is described in.

There remains a medical need for new treatments for stroke and cerebrovascular disorders. The aim of the present invention is to provide such a treatment.

It has now been discovered that certain azetidine-1-carboxamides show unexpected neuroprotective efficacy as compared to standard GABAergic agents. In particular, certain azetidine-1-carboxamides have been shown to potentiate the action of GABA in inhibiting neurons, and prevent repetitive firing induced as a result of activation of glutamatergic mechanisms. Was shown to do. Such compounds are neuroprotective after acute cerebral ischemia in rats and mice, and arechemia-induced CNS in an in vivo model of focal ischemia in both species.
It was found to reduce damage.

According to the present invention, in the manufacture of a medicament for neuroprotection or treatment of cerebral ischemia, central nervous system injury or ocular disease in a subject, the following:

[0023]

[Chemical 2] Use of a compound of formula (I), or a pharmaceutically acceptable salt or prodrug thereof, wherein: R ¹ is aryl; R ² is H, alkyl or aryl; R ³ is hydrogen or alkyl.

[0024]   Reference herein to an “alkyl” group refers to a branched or unbranched ring
Formula or acyclic, saturated or unsaturated (eg, alkenyl (including allyl))
Or alkynyl (including propargyl)) hydrocarbyl group. here
And the cyclic or acyclic alkyl group is preferably C₁~ C₁₂And more preferred
It is C₁~ C₈(For example, methyl, ethyl, propyl, isopropyl, butyl,
Isobutyl, tert-butyl, pentyl, isopentyl, hexyl, heptyl
, Octyl). Therefore, the term "alkyl" as used herein
, Alkyl (branched or unbranched), alkenyl (branched or unbranched)
, Alkynyl (branched or unbranched), cycloalkyl, cycloalkenyl and
And cycloalkynyl. In the preferred embodiment
And the cyclic alkyl group is preferably C₃~ C₁₂And more preferably C_Five~ C ₈ And the acyclic alkyl group is preferably C₁~ C_TenAnd more preferred
Ku, C₁~ C₆And more preferably methyl, ethyl, propyl (n-pro
Pill or isopropyl), butyl (n-butyl, isobutyl or tert)
-Butyl) or pentyl (including n-pentyl and iso-pentyl)
, And more preferably methyl.

Reference herein to an “aryl” group refers to a monocyclic or bicyclic aromatic group (eg, phenyl or naphthyl).

The alkyl and aryl groups may be substituted or unsubstituted. In one embodiment, only the alkyl and aryl groups defined above as R _{1 to} R ₃ can be substituted. When substituted, there will generally be 1 to 3 substituents present, preferably 1 or 2 substituents. Substituents include the following: carbon-containing groups such as alkyl, aryl, arylalkyl (eg, substituted and unsubstituted phenyl, substituted and unsubstituted benzyl); hydrogen atoms and halogen-containing groups such as haloalkyl (eg, tri Fluoromethyl); alcohols (eg hydroxy, hydroxyalkyl, (aryl) (hydroxy) alkyl), ethers (eg alkoxy, alkoxyalkyl, aryloxyalkyl), aldehydes (eg carboxaldehyde), ketones (eg alkylcarbonyl) Acid derivatives (eg, alkoxy, alkylcarbonylalkyl, arylcarbonyl, arylalkylcarbonyl, arylcarbonylalkyl), acids (eg, carboxy, carboxyalkyl), esters (eg, alkoxy) Rubonyl, alkoxycarbonylalkyl, alkylcarbonyloxy, alkylcarbonyloxyalkyl), and amido (eg, aminocarbonyl, monoalkylaminocarbonyl or dialkylaminocarbonyl, aminocarbonylalkyl, monoalkylaminocarbonylalkyl or dialkylaminocarbonylalkyl, arylamino) An oxygen-containing group such as carbonyl); an amine (eg, amino, monoalkylamino or dialkylamino, aminoalkyl, monoalkylaminoalkyl or dialkylaminoalkyl),
Nitrogen-containing groups such as azides, nitriles (eg cyano, cyanoalkyl), nitro; thiols, thioesters, sulfoxides and sulfones (eg alkylthio, alkylsulfinyl, alkylsulfonyl, alkylthioalkyl, alkylsulfinylalkyl, alkylsulfonylalkyl, arylthio). , A sulfur-containing group such as arylsulfinyl, arylsulfonyl, arylthioalkyl, arylsulfinylalkyl, arylsulfonylalkyl); and a heterocyclic group containing one or more, preferably one heteroatom (eg thienyl, furanyl). , Pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazoli Nyl, tetrahydrofuranyl, pyranyl, pyronyl, pyridyl, pyrazinyl, pyridazinyl, piperidyl, piperazinyl, morpholinyl, thionaphthyl, benzofuranyl, isobenzofuryl, indolyl, oxyindolyl, isoindolyl, indazolyl, indolinyl, 7-azaindolyl, isoindazolyl, benzopyranyl, coumarinyl , Isocoumarinyl, quinolyl, isoquinolyl, naphthyridinyl, cinnolinyl, quinazolinyl, pyridopyridyl, benzoxazinyl, quinoxazinyl, chromenyl, chromanyl, isochromanyl and carborinyl).

Preferred substituents include halogen-containing groups such as alkyl, aryl, nitrile, halo, or trifluoromethyl.

As used herein, the term “alkoxy” means alkyl-O— and “alcoyl” means alkyl-CO—.

As used herein, the term “halogen” means a fluorine radical, a chlorine radical, a bromine radical, or an iodine radical, preferably a fluorine radical or a chlorine radical.

The compounds of formula (I) may exist in many diastereomeric and / or enantiomeric forms. Unless otherwise stated, references herein to "a compound of formula (I)" are to all stereoisomeric forms of the compound and to the unseparated stereoisomers ( Racemic or non-racemic), and each stereoisomer in its pure form.

In a preferred embodiment of the invention, the compound of formula (I) is a compound of formula (I)
R) -enantiomer (substantially free of its (S) -enantiomer).

In the compounds of formula (I), preferred R ¹ is substituted or unsubstituted phenyl or naphthyl, more preferred R ¹ is substituted phenyl or naphthyl, more preferred R ¹ is 1 Is phenyl or naphthyl with 3 to 3 substituents, and the most preferred R ¹ is phenyl or naphthyl with 1 or 2 substituents. When R ¹ is phenyl with one substituent, the phenyl group is preferably para- or meta-substituted. If R ¹ is phenyl with two substituents, this phenyl group is preferably substituted in the meta and para positions. Most preferred R ¹ groups are 4-chlorophenyl, 4-
It is selected from fluorophenyl, 3-trifluoromethylphenyl, 3,4-dichlorophenyl, and 3,4-difluorophenyl.

In compounds of formula (I), preferred R ² is H or alkyl, more preferred R ² is H or acyclic hydrocarbyl, more preferred R ² is H
Or methyl, and the most preferred R ² is H.

In one embodiment of the invention, in the compound of formula (I) R ³ is alkyl, preferably alkenyl, alkynyl, hydroxyalkyl, alkoxyalkyl, or unsubstituted saturated cyclic or unsubstituted saturated non- Cyclic, hydrocarbyl, and more preferably allyl or propargyl.

[0035]   Particularly preferred compounds are:

[0036]

[Chemical 3] Among these, preferred compounds are 3- (3,4-dichlorobenzyloxy)
-N- (2-propenyl) azetidine-1-carboxamide, 3- (3- (trifluoromethyl) benzyloxy) -N- (2-propenyl) azetidine-1-
Carboxamide, 3- (4- (trifluoromethyl) benzyloxy) -N- (
2-propenyl) azetidine-1-carboxamide, 3- (4-fluorobenzyloxy) -N- (2-propenyl) azetidine-1-carboxamide, 3- (
Bis (4-chlorophenyl) methoxy) -N- (2-propenyl) azetidine-
1-carboxamide, (R) -3- (bis (4-chlorophenyl) methoxy)-
N- (2-hydroxypropyl) azetidine-1-carboxamide, 3-((3
-Chlorophenyl) methoxy) -azetidine-1-carboxamide, and 3-
(1- (3-trifluoromethylphenyl) ethyloxy) -azetidine-1-
It is a carboxamide.

According to a further aspect of the invention, comprising administering an effective dose of a compound of formula (I), or a pharmaceutically acceptable salt thereof or a prodrug thereof, to a subject in need of such treatment. A method of neuroprotection is provided.

According to a further aspect of the invention, the method comprises administering to a subject in need of such treatment an effective dose of a compound of formula (I), or a pharmaceutically acceptable salt thereof or a prodrug thereof. A method of treating cerebral ischemia, central nervous system injury, or eye disease is provided.

The present invention may be used with respect to human or animal subjects, more preferably mammalian subjects, more preferably human subjects.

[0040]   As used herein, the term "treatment" includes prophylactic treatment.

The term “prodrug”, as used herein, means any pharmaceutically acceptable prodrug of a compound of formula (I). For example, a compound of formula (I) may be prepared in a prodrug, wherein the free -OH group is a suitable group (eg, alkyl, aryl, phosphate, sugar, amine, glycol, sulfonate). , Or may include an acid functional group) (eg, via an ester bond, an amide bond, or a phosphate bond). This suitable group will eventually become (e.g., when, after administration or when exposed to the desired biological environment)
Suitable instability such that it is removed / cleaved (by hydrolysis) to expose the compound of formula (I).

As used herein, the term “pharmaceutically acceptable salt” means any pharmaceutically acceptable prodrug of a compound of formula (I). Salts are prepared from pharmaceutically acceptable non-toxic acids and bases including inorganic and organic acids and bases. Such acids include the following: acetic acid, benzenesulfonic acid, benzoic acid,
Camphor sulfonic acid, citric acid, ethanesulfonic acid, dichloroacetic acid, fumaric acid, gluconic acid, glutamic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, nitric acid, pamo ( acid, pantothenic acid, phosphoric acid, succinic acid, methanesulfonic acid, mucus acid, sulfuric acid, tartaric acid, oxalic acid, p
-Toluenesulfonic acid and the like. Particularly preferred are hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and methanesulphonic acid, and most particularly preferred is methanesulphonate. Acceptable base salts include alkali metal salts (eg, sodium, potassium), alkaline earth metal salts (eg, calcium, magnesium), and aluminum salts.

As used herein, the term “substantially free of its (S) -enantiomer” refers to a pharmaceutical or therapeutic composition comprising a compound of formula (I) used according to the invention. , Containing a higher proportion of the (R) -enantiomer of the compound of formula (I) relative to the (S) -enantiomer of the compound of formula (I). In a preferred embodiment of the invention, the term "substantially free of its (S) -enantiomer" as used herein means that the composition comprises at least 90% by weight of the (R) -enantiomer. , And 10% by weight or less of the (S) -enantiomer. In a further preferred embodiment, the term "substantially free of its (S) -enantiomer" means that the composition comprises at least 99% by weight of the (R) -enantiomer and 1% by weight of the (S) -enantiomer. It is meant to include: In another preferred embodiment, the term "substantially free of its (S) -enantiomer" means that the composition comprises 100% by weight of the (R) enantiomer. The above proportions are based on the total amount of compound of formula (I) present in the pharmaceutical or therapeutic composition used according to the invention.

Diseases, disorders, and medical procedures / procedures relating to the present invention are: cerebral ischemia, which is a transient cerebral ischemic attack, stroke (thrombotic stroke, ischemic stroke, embolic stroke). , Hemorrhagic stroke, hiatal stroke), subarachnoid hemorrhage, cerebral vasospasm, neuroprotection for stroke, peri-natal asphyxia, drowning, carbon monoxide poisoning, cardiac arrest and subdural hematoma. Central nervous system injury, which includes traumatic brain injury, neurosurgery (surgical injury), neuroprotection for head injury, increased intracranial pressure, cerebral edema, hydrocephalus and spinal cord injury. And ocular diseases, which include drug-induced optic neuritis, cataracts, diabetic neuropathy, ischemic retinopathy, retinal hemorrhage, retinitis pigmentosa, acute glaucoma, chronic glaucoma, macular degeneration, retinal artery occlusion and retinitis. Including ).

In addition, compounds of formula (I) may also be used to potentiate the effects of other treatments (eg, enhance neuroprotective effects of the nerve growth factor-derived brain).

The present invention specifically relates to the treatment of cerebral ischemia and central nervous system injury. The present invention is also specifically directed to post-asphyxia brain injury (post-asph) in neonatal subjects.
yxial brain damage).

The compound of formula (I) may be one or more additional drugs useful in the treatment of the disorders described above, the components of which may be in the same formulation or in separate formulations for simultaneous or sequential administration. Present).

Compounds of formula (I) may be prepared according to the reaction scheme, where P is a nitrogen protecting group. R ¹ , R ² and R ³ have already been defined. Ether (IV) is an aryl alkanol (III, X = OH) of azetidinol (II) and diethyl azodicarboxylate and triphenylphosphine, or an arylalkyl chloride, an arylalkyl bromide, an arylalkyliodine, an arylalkyl mesylate. , Or arylalkyl tosylate (I
II, X = Cl, Br, I, mesylate, tosylate) and a strong base (eg,
It can be formed by reaction with either sodium hydroxide). Azetidine (V)
Can be achieved by reaction of (IV) with a suitable nitrogen deprotecting agent. For example, if P is a diphenylmethyl group, deprotection can be carried out by treatment with 1-chloroethyl chloroformate followed by methanol. Urea (I) is formed by the reaction of azetidine (V) with an N-alkyl isocyanate or N-alkyl carbamoyl chloride and a base (eg triethylamine or potassium carbonate). Alternatively, urea is an intermediate (eg secondary amine (V))
Can be prepared directly from azetidine (IV) without isolation. For example, when P is a diphenylmethyl group, azetidine (IV) is added to phosgene followed by amine R ³
Urea (I) can be obtained directly by treatment with NH ₂ .

[0049]   (Reaction scheme)

[0050]

[Chemical 4] The present invention provides a pharmaceutical composition comprising an effective amount of a compound of formula (I) in combination with a pharmaceutically acceptable carrier or excipient, and a pharmaceutically acceptable amount of the compound of formula (I). Further provided are methods of making such compositions, including the step of combining with possible carriers or excipients.

To further increase potency, the composition may include ingredients such as dextran or cyclodextrins, or their ether derivatives, which help stabilize and disperse the active ingredient and metabolize the active ingredient. Lower.

For compositions where the pharmaceutically acceptable carrier comprises cyclodextrin or an ether derivative thereof, the active ingredient is intimately mixed with an aqueous solution of cyclodextrin or an ether derivative thereof. This optionally involves the addition of additional pharmaceutically acceptable ingredients before, during, or after this mixing. Therefore, the resulting solution is optionally lyophilized and the lyophilized residue is optionally reconstituted with water.

In one embodiment of the invention, the composition further comprises a buffer system, a tonicity agent and water.

The compounds of formula (I) may be administered in a form suitable for: oral use, eg
Tablets, capsules, aqueous or oily solutions, suspensions or emulsions; topical applications including transmucosal and transdermal applications, eg creams, ointments
nt), gel, aqueous or oily solution or suspension, salve
, Patches or plasters; nasal use, eg snuff, nasal spray or nasal drops; vaginal or rectal use, eg suppositories; administration by inhalation, eg fine or liquid aerosols; sublingual or buccal Uses, eg tablets or capsules; or parenteral uses (including intravenous, subcutaneous, intramuscular, intravascular or infusion), eg
Sterile water or oily solution or suspension. In general, the compositions described above may be prepared in a conventional manner using conventional excipients, using standard techniques well known to those of ordinary skill in the pharmaceutical arts. Preferably the compound is administered orally.

For oral administration, the compounds of formula (I) will generally be provided in the form of tablets or capsules or as an aqueous solution or suspension.

Tablets for oral use are mixed with pharmaceutically acceptable excipients such as inert diluents, disintegrating agents, binders, lubricants, flavoring agents, coloring agents and preservatives. The active ingredients mentioned may be mentioned. Suitable inert diluents include sodium and potassium carbonate, sodium and calcium phosphates, and lactose, while corn starch and alginic acid are suitable disintegrating agents. Binders may include starch and gelatin, while, if any,
The lubricant is generally sodium stearate, stearic acid or talc.
If desired, the tablets may be coated with a substance such as glycerol monostearate or glyceryl distearate to delay absorption in the gastrointestinal tract.

Capsules for oral use include hard gelatin capsules, in which the active ingredient is mixed with a solid diluent, and soft gelatin capsules, where
Active ingredients include water or oils (eg mixed with peanut oil, liquid paraffin or olive oil).

For intramuscular, intraperitoneal, subcutaneous and intravenous applications, the compounds of formula (I) are generally provided in a sterile aqueous solution or suspension and buffered to a suitable pH and isotonicity. Be converted. Suitable aqueous vehicles include Ringer's solution and isotonic sodium chloride. Aqueous suspensions may include suspending agents such as cellulose derivatives, sodium alginate, polyvinyl-pyrrolidone and gum tragacanth, and wetting agents such as lecithin. Suitable preservatives for aqueous suspensions include ethyl p-hydroxybenzoate and n-propyl p-hydroxybenzoate.

It is understood that the dosage levels used can vary within fairly wide ranges depending on the compound used, the severity of the symptoms exhibited by the patient, and the weight of the patient.

The present invention is described herein in detail with reference to the following pharmacological examples. It is understood that this example is intended to illustrate the invention and not to limit the scope of the invention.

(Example) (Synthetic Example) (Preparation of 1- (diphenylmethyl) -3-azetidinol) This compound was prepared according to Anderson and Lok (J. Org. Chem. 19).
72, 37, 3953, the disclosure of which is incorporated herein by reference. m. p. 111-112 ° C (lit.mp 113 ° C).

(Preparation of 3- (4-chlorobenzyloxy) -1- (diphenylmethyl) azetidine (1)) 1-Diphenylmethyl-3-azetidinol (25 in DMF (100 mL).
mmol) solution at 0 ° C. in NaF (60% in oil) in DMF (50 mL).
Dispersion, 30 mmol) was added to the suspension. The reaction mixture was stirred for 1 hour at room temperature, then 4-chlorobenzyl chloride (25 mmol) was added dropwise at 0 ° C. and the reaction mixture was stirred for 3 hours at room temperature. The reaction was quenched with water and extracted with ethyl acetate (3 × 50 mL), and the extract was washed with water and brine, dried (MgSO _4), and concentrated under reduced pressure. The residue was purified by chromatography [SiO ₂ ; hexane-ethyl acetate (9: 1)] to give the product as a yellow oil (7.3 g, 80%). This material was used in the next step without further purification.

Example 1. 3- (4-chlorobenzyloxy) -N- (2-propenyl)
To a solution of azetidine-1-carboxamide _{(2)) CH 2 Cl 2} (40mL) compounds in (1) (20 mmol), at 0 ° C., it was added phosgene solution (in toluene 1.75 M, 24 mmol). The reaction mixture was stirred at room temperature for 90 minutes, concentrated under reduced pressure and then CH ₂ Cl ₂ (40 mL).
) And treated with allylamine (42 mmol) at 0 ° C. The reaction was stirred at room temperature for 4 hours, then water (40 mL) was added and the phases were separated. The aqueous phase was extracted with additional _{CH 2 Cl 2 (2 × 40mL} ). The organic phase is diluted with HCl (
20 mmol) and brine, dried (MgSO _4), and concentrated under reduced pressure. The residue was triturated with diethyl ether to give product (2) as a crystalline solid. (3.5 g, 60%). m. p. 110-111 ° C.
Found: C, 59.84; H, 6.11; N, 9.98. C ₁₄ H ₁₇ ClN ₂ O ₂ Calculated: C, 59.89; H, 9.6.10 ; N, 9.97%.

Preparation of 3- (3,4-dichlorobenzyloxy) -1- (diphenylmethyl) azetidine (3) This material was prepared from 1-diphenyl using the procedure described for compound (1). Prepared from methyl-3-azetidinol (6.0 g) and α, 3,4-trichlorotoluene (yield 92%).

Example 2. 3- (3,4-Dichlorobenzyloxy) -N- (2-propenyl) azetidine-1-carboxamide (4) This material was prepared according to the procedure described for compound (2). Using the compound (3)
Prepared from (9.2 g) (75% yield). m. p. 88-89 ° C. Measured value: C
, 53.43; H, 5.18; N, 8.85. Calculated for C ₁₄ H ₁₆ Cl ₂ N ₂ O ₂ :
C53.3; H, 5.12; N, 8.88%.

Preparation of 3- (3-Trifluoromethyl) benzyloxy) -1- (diphenylmethyl) azetidine (5) This material was prepared using the procedure described for compound (1) to 1- Prepared from diphenylmethyl-3-azetidinol (5 g) and α'-bromo-α, α, α-trifluoro-m-xylene (91% yield).

Example 3. 3- (3- (Trifluoromethyl) benzyloxy) -N- (
2-Propenyl) azetidine-1-carboxamide (6)) This material was converted to compound (5) using the procedure described for compound (1).
(7.5 g). m. p. 108 ° C. Found: C, 57.29; H,
5.44; N, 8.87. Calculated for C ₁₅ H ₁₇ F ₃ N ₂ O ₂ : C, 57.32; H,
5.45; N, 8.91%.

Preparation of 3- (4- (trifluoromethyl) benzyloxy) -1- (diphenylmethyl) azetidine (7) This material was prepared using the procedure described for compound (1) to give 1 -Diphenylmethyl-3-azetidinol (6.0 g) and α'-bromo-α, α, α-
Prepared from trifluoro-p-xylene (77% yield).

Example 4. 3- (4- (trifluoromethyl) benzyloxy) -N- (
2-Propenyl) azetidine-1-carboxamide (8)) This material was treated with the compound (7) using the procedure described for compound (2).
n (yield 72%). m. p. 120 ° C. Found: C, 57.27;
H, 5.45; N, 8.86. Calculated for C ₁₅ H ₁₇ F ₃ N ₂ O ₂ : C, 57.32.
H, 5.45; N, 8.91%.

Preparation of 3- (4-Fluorobenzyloxy) -1- (diphenylmethyl) azetidine (9) This material was prepared using the procedure described for compound C1) in 1-diphenylmethyl-3-. Prepared from azetidinol (6.0 g) and 4-fluorobenzyl bromide (83% yield).

Example 5. 3- (4-Fluorobenzyloxy) -N- (2-propenyl) azetidine-1-carboxamide (10) This material was prepared using the procedure described for compound (2). And compound (9)
Prepared from. m. p. 97-99 ° C. Found: C, 63.57; H, 6.59.
N, 10.66. Calculated for C ₁₄ H ₁₇ ClN ₂ O ₂ : C, 63.62; H, 6.4.
8; N, 10.59%.

(Preparation of 3- (bis- (4-chlorophenyl) methoxy-1-diphenylmethyl) azetidine (11)) 4,4′-dichlorobenzhydrol (25 m) in benzene (100 mL)
mol), p-toluenesulfonic acid (18.4 mmol), and 1- (diphenylmethyl) -3-azetidinol (8.4 mmol) in Dean-S.
Heated to reflux in the tark apparatus for 3 hours. The solution was cooled, washed with sodium bicarbonate (saturated aqueous solution, 100 mL), dried (MgSO _4), and concentrated under reduced pressure. The residue was purified by chromatography [SiO ₂ ; hexane-diethyl ether (5: 1)] to give the product (11) as a cloudy oil which crystallized on standing (2.4 g, 62%).

Example 6. 3- (Bis (4-chlorophenyl) methoxy) -N- (2-propenyl) azetidine-1-carboxamide (12) This material was prepared according to the procedure described for compound (2). Use the compound (11
) Was prepared as a crystalline solid. Found: C, 56.38; H, 5.10.
N, 6.51. _{C 20 H 20 Cl 2 N 2} O 2 · 2H 2 O Calculated: C, 56.21; H
, 5.66; N, 6.56%.

Example 7. (R) -3- (bis (4-chlorophenyl) methoxy) -N-
Preparation of (2-hydroxypropyl) azetidine-1-carboxamide (13) This material was prepared using the procedure described for compound (2) to give compound (11
) And (R)-(-)-1-amino-2-propanol as a crystalline solid (yield 57%). Found: C, 58.74; H, 5.42; N, 6
． 84. Calculated for _{C 20 H 22 Cl 2 N 2} O 3: C, 58.69; H, 5.42; N,
6.84%.

Example 8. 3- (3-Trifluoromethyl) benzyloxy-N-azetidine-1-carboxamide (14) 3- (3-trifluoromethyl) benzyloxy-1 in dichloromethane (15 mL). To a solution of-(diphenylmethyl) azetidine (5) (5.3 mmol) at 0 <0> C was added a solution of phosgene (1.75M in toluene, 6.4 mmol). The reaction mixture was stirred at room temperature for 2 hours, concentrated under reduced pressure and then THF (1
Aluminum hydroxide (5 mL) dissolved in 5 mL) and added at 0 ° C. in one portion
Processed in. The reaction was stirred vigorously at room temperature for 15 hours, then water (50 mL).
) And ethyl acetate (40 mL) were added and the phases were separated. The aqueous phase was extracted with ethyl acetate (2 x 40 mL), dried (MgSO4) and concentrated under reduced pressure. The residue was triturated with ethyl acetate (10 mL) to give (14) as a solid (0.91 g, 63%). mp. 167 ° C (ethyl acetate). Measured value: C
, 52.44; H4.72; N, 10.23. Calculated value for C ₁₄ H ₁₇ ClN ₂ O ₂ : C
, 52.56; H, 4.78; N, 10.21.

(Preparation of 3- (1- (3-trifluoromethylphenyl) ethyloxy) -1- (diphenylmethyl) azetidine (15)) α-Methyl-3-trifluoromethylbenzyl in dichloromethane (150 mL) Alcohol (53 mmol), diisopropylethylamine (105 mm
sol) under nitrogen and cooled to 0 ° C. to give methanesulfonyl chloride (6
(3.1 mmol) was added dropwise over 10 minutes. The reaction was stirred for 15 hours.
Water (200 mL) was added and the resulting mixture was stirred for 10 minutes, poured into potassium carbonate (10% wt / wt aqueous solution, 200 mL) and extracted with dichloromethane (3 x 150 ml). The combined organic extracts were washed once with brine (50 mL) then dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The residue was dissolved in ethyl ether and washed through a pad of silica, eluting with more ether. The filtrate was concentrated under reduced pressure. This material was used directly, as shown below.

A solution of 1-diphenylmethyl-3-azetidinol (42 mmol) in dimethylformamide (20 mL) was added at 0 ° C. to dimethylformamide (80 mL).
To a suspension of NaH (60% dispersion in oil, 50 mmol) in was pipetted. The reaction mixture was stirred at room temperature for 15 minutes and the above crude material (assuming 53 mmol) was added dropwise at 0 ° C. as a solution in dimethylformamide (30 mL).
The reaction mixture was then stirred at room temperature for 2 hours. Water the reaction (200 mL)
Pour into and extract with ethyl acetate (3 x 50 mL) and extract this with water (2
Washed with 200 mL) and brine (50 mL), dried (MgSO _4), and concentrated under reduced pressure. The residue is chromatographed (SiO ₂ ; hexane /
Purification with ethyl acetate 9/1) gave 3- (1- (3-trifluoromethylphenyl) ethyloxy) -1- (diphenylmethyl) azetidine (15) as a yellow oil (11.2 g, Yield 65%). This material was used in the next step without further purification.

Example 9. 3- (1- (3-Trifluoromethylphenyl) ethyloxy) -azetidine-1-carboxamide (16) This material was prepared using the procedure described for compound (14). , Compound (1
Prepared from 5) as a crystalline solid (yield 62%). mp. 130.5-1
31.5 ° C (diisopropyl ether). Found: C, 54.24; H, 5.2.
6; N, 9.69. C ₁₄ H ₁₇ ClN ₂ O ₂ Calculated: C, 54.17; H, 5.2
4; N, 9.71.

The individual enantiomers of Example 9 were used except that the chiral alcohol was used.
Prepared using the same total synthetic method as described for compound 16. The R-enantiomer of Example 9 was converted to the appropriate chiral 1- (3-trifluoromethyl)
Prepared from phenylethyl alcohol. These chiral alcohols could be prepared from 3'-trifluoromethyl-acetophenone by stereoselective reduction using, for example, borates and suitable chiral auxiliaries or chiral catalysts (Corey, EJ; Bakshi. , RK; Shibata S. J. Amer.
． Chem. Soc. , 1987, 109, 5551-5553, or Pic
kard, ST and Smith, HE. J. Amer. Chem. Soc. ，
1990, 112, 5741-5747).

Examples 10-43-See Table 1 These products were prepared using the procedure described for compound (2).

[0081]

[Table 1] (Footnote of Table 1) Footnote a: 3296, 2980, 2943, 2877, 1638, 1545, 14
00, 1377, 1330, 1203, 1166, 1127, 1073, 706
cm ^-1 . Footnote b: 3319, 2963, 2872, 1634, 1549, 1469, 14
03, 1327, 1269, 1184, 1130, 1083, 818 cm ^-1 . Example 44. 3-((3-Chlorophenyl) methoxy) -azetidine-1
-Carboxamide (51)) This material was treated with compound (1) using the procedure described for compound (14).
) Was prepared as a crystalline solid (yield 87%). m. p. 163-165
． 5 ° C (diisopropyl ether). Found: C, 55.49; H, 5.45;
N, 11.40. Calculated for C ₁₁ H ₁₃ ClN ₂ O ₂ : C, 54.89; H, 5.44.
N, 11.63.

Test Procedures Rat Transient Middle Cerebral Artery Occlusion (MCAo) Ischemia Model This model of middle cerebral artery occlusion used is based on intravascular microfiber technology in rats (Zhao Q. et al., Acta. Physiol. Scand. 199
4,152,349-350). Male Lister Hooded rats were used for these experiments and divided into 3 groups (Group 1: Vehicle; Group 2: Clomethiazole (CMZ); Group 3: Compounds of Formula (I)). The sample size of each was 11-15. The animal was anesthetized and the carotid artery was exposed. Introduce a percutaneous suture (3/0) with a specific diameter into the ligated carotid artery and pass it through the external carotid artery and common carotid bifurcation, internal carotid artery and pterygopalatine artery for cranial circulation. Introduced. The fibrils were then retained in the narrow proximal anterior carotid artery to occlude the middle cerebral artery. 90 minutes after occlusion of the middle cerebral artery, fibrils were removed and recirculated.

22.5 hours after reperfusion, the animals were perfused via the transaortic route with 200 ml of a 4% solution of tetrazolium chloride warmed to 37 ° C. After perfusion,
Brains were removed and soaked and fixed in 10% formalin / saline for at least 48 hours. After fixation, the brain was sliced into 0.5 mm sections with vibrosulile. Using this technique, viable tissue was stained dark red and infarcted tissue was not stained. The infarct area of each section was measured and the total infarct volume in the cerebral hemisphere, cortex and striatum was calculated using the Kontron image analysis system.

(Mouse Permanent Middle Cerebral Artery Occlusion Ischemia Model) Adult male C57B1 mice (20) were prepared 30 minutes before middle cerebral artery (MCA) occlusion.
˜25 g, n = 10 / group) were intraperitoneally administered compound of formula (I) (10 mg / kg) or vehicle (60% PEG400 in water). Under halothane anesthesia (1.5% halothane in nitrous oxide: oxygen (70:30)) a small local cranial resection was performed
The left MCA was exposed. The distal portion of the MCA was occluded by electrocoagulation. The incision site was sutured and the anesthesia was stopped. Twenty-four hours after MCA occlusion, the mice were euthanized, brains removed and immersed in 4% tetrazolium chloride to visualize areas of infarction (Backhaus C. et al., J. Pha.
rm Methods 1992, 27, 27-32). The brain was then stored in 10% formalin / saline. Then, the area of visible infarct formation on the surface of the cortex was measured by the PC digital imaging system (KS300, Imaging A).
Calculated using Associates, UK). The data generated is the absolute area of infarct formation (mm ² ) in each animal. Mean cerebral infarct area was compared by a one-tailed t-test with the significance taken at p <0.05.

Experimental results are shown in FIGS. 1 and 2, which show (i) vehicle; and (i
i) shows the effect of the compound of formula (I) on infarct formation after permanent middle cerebral artery occlusion.

[Brief description of drawings]

FIG. 1 shows that the R-enantiomer of Compound 16 shows significant neuroprotection at an intraperitoneal dose of 60 mg / kg when administered 30 minutes before occlusion in a mouse permanent MCAo model. Indicates.

FIG. 2 shows that in a mouse permanent MCAo model, compound 51 shows significant neuroprotection at an intraperitoneal dose of 100 mg / kg when administered concurrently with occlusion.

─────────────────────────────────────────────────── ─── Continued front page    (81) Designated countries EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, I T, LU, MC, NL, PT, SE), OA (BF, BJ , CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, K E, 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, C A, CH, CN, CR, CU, CZ, DE, DK, DM , DZ, EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, K E, KG, KP, KR, KZ, LC, LK, LR, LS , LT, LU, LV, MA, MD, MG, MK, MN, MW, MX, MZ, NO, NZ, PL, PT, RO, R U, SD, SE, SG, SI, SK, SL, TJ, TM , TR, TT, TZ, UA, UG, US, UZ, VN, YU, ZA, ZW (72) Inventor Monk, Nathaniel Julius             UK Country RG 41 5 UAE             Wokingham, Winnersh, Lee             Ding Road 613, Oakden             coat (72) Inventor Fret Char, Alan             UK Country RG 41 5 UAE             Wokingham, Winnersh, Lee             Ding Road 613, Oakden             coat (72) Inventor Stanhope, Kelly Jean             UK Country RG 41 5 UAE             Wokingham, Winnersh, Lee             Ding Road 613, Oakden             coat (72) Inventor Munsell, Howard Lanham             UK Country RG 41 5 UAE             Wokingham, Winnersh, Lee             Ding Road 613, Oakden             coat (72) Inventor Nelson, Alan John             UK Country RG 41 5 UAE             Wokingham, Winnersh, Lee             Ding Road 613, Oakden             coat F-term (reference) 4C086 AA01 AA02 BC04 MA01 MA04                       NA14 ZA01 ZA02 ZA15 ZA33                       ZA36

Claims (17)

[Claims] 1. A compound of formula (I), or a pharmaceutically acceptable compound thereof, in the manufacture of a medicament for neuroprotection in a subject or for the treatment of cerebral ischemia, central nervous system injury or ocular disease. Use of salts or prodrugs: Wherein R ¹ is aryl; R ² is H, alkyl or aryl; and R ³ is hydrogen or alkyl. 2. The use according to claim 1, wherein R ¹ is substituted or unsubstituted phenyl or naphthyl. 3. Use according to claim 1 or 2, wherein R ¹ has ¹ , 2 or 3 substituents. 4. Use according to claim 1, 2 or 3, wherein R ¹ is chlorophenyl, fluorophenyl, (trifluoromethyl) phenyl, 3,4-dichlorophenyl or 3,4-difluorophenyl. 5. Use according to claim 1, 2, 3 or 4, wherein R ² is hydrogen or methyl. It is wherein R ^3, are alkyl, Use according to any one of claims 1 to 5. 7. Use according to any one of claims 1 to 5, wherein R ³ is alkenyl, alkynyl, hydroxyalkyl or alkoxyalkyl. 8. The method according to claim 1, wherein R ³ is alkyl or propargyl.
The use according to any one of 1. 9. Use according to any one of claims 1 to 5, wherein R ³ is an unsubstituted saturated cyclic or acyclic hydrocarbyl. 10. The compound is 3- (4-chlorobenzyloxy) -N- (2-propenyl) azetidine-1.
-Carboxamide, 3- (3,4-dichlorobenzyloxy) -N- (2-propenyl) azetidine-1-carboxamide, 3- (3- (trifluoromethyl) benzyloxy) -N- (2-propenyl) azetidine -1-carboxamide, 3- (4- (trifluoromethyl) benzyloxy) -N- (2-propenyl) azetidine-1-carboxamide, 3- (4-fluorobenzyloxy) -N- (2-propenyl) azetidine −
1-carboxamide, 3- (bis (4-chlorophenyl) methoxy) -N- (2-propenyl) azetidine-1-carboxamide, (R) -3- (bis (4-chlorophenyl) methoxy) -N- (2- Selected from hydroxypropyl) azetidine-1-carboxamide, 3-((3-chlorophenyl) methoxy) -azetidine-1-carboxamide, and 3- (1- (3-trifluoromethylphenyl) ethyloxy) -azetidine-1-carboxamide. The use according to claim 1, which is carried out. 11. Use according to any one of claims 1 to 10, wherein the medicament comprises a pharmaceutically acceptable carrier and, as an active ingredient, an effective amount of a compound of formula (I). 12. The use according to claim 11, wherein the carrier comprises cyclodextrin or an ether derivative thereof. 13. The use according to any one of claims 1 to 12, wherein the medicament further comprises a buffer system, an isotonicity agent and water. 14. Use according to any one of claims 1 to 13, wherein the compound of formula (I) is in neuroprotection or for the treatment of cerebral ischemia, central nervous system injury or eye diseases. In combination with one or more additional drugs useful in, the compound is the same formulation or separate formulations for simultaneous or sequential administration. 15. A method of neuroprotection, which method comprises administering to a subject in need of such treatment an effective dose of a compound of formula (I) as defined in any one of claims 1-10. A method comprising administering a compound, or a pharmaceutically acceptable salt or prodrug thereof. 16. A method of treating cerebral ischemia, central nervous system injury or eye disease, which method comprises administering to a subject in need of such treatment an effective dose.
0. A method comprising the step of administering a compound of formula (I) as defined in any one of 0, or a pharmaceutically acceptable salt or prodrug thereof. 17. The method according to claim 15 or 16, wherein the compound of formula (I) is administered in the form as defined in any one of claims 11, 12, 13 or 14.