WO2008109286A1 - Methods for treating cognitive disorders using 1-aryl-1-hydroxy-2,3-diamino-propyl amines, 1-heteroaryl-1-hydroxy-2,3-diamino-propyl amines and related compounds - Google Patents

Abstract

Disclosed herein are methods of treating a patient suffering from a cognitive disorder using compounds of the following formula, Formula (2), wherein the variables have the meaning defined in the specification.

Description

Docket 18174PCT (AP)

METHODS FOR TREΞATING COGNITIVE DISORDERS USING 1-ARYL-1-

HYDROXY-2,3-DIAMINO-PROPYL AMINES, 1-HETEROARYL-1 -HYDROXY-

2,3-DIAMINO-PROPYL AMINES AND RELATED COMPOUNDS

CROSS REFERENCE TO RELATED APPLICATIONS

This application is based on, and claims the benefit of, U S Provisional Application No 60/893,203, filed March 6, 2007, and which is incorporated herein by reference

BACKGROUND OF THE INVENTION Field of the Invention

The present invention is directed to methods of treating a patient suffering from one or more types of cognitive disorders using derivatives of 1- aryl-1-hydroxy-2,3-dιamιno-propyl amines, 1-heteroaryl-1-hydroxy-2,3- diamino-propyl amines and related compounds Background Art i-Phenyl^-decanoylamino-S-morpholino-i-propanol (PDMP) was discovered by Vunam, R R and Radin, N , Chem Phys Lipids, 26, 265-278, 1980 Preparation of PDMP is described in Inokuchi, J et al , J Lipid Res 28, 565-571 , 1987, Radin, A er a/, NeuroProtocols, 3(2), 145-55, 1993, Abe er a/, J Lipid Res 36, 611-621, 1995 and US 5916911

PDNIP mixture of DL-βrythro and DL-threo isomers

The isomers most active have the R,R-(D-fΛreo)-confιguratιon 2008/054936

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Preparation of enantiomerically pure D-f/?reo-PDMP has been reported by Mitchell, Scott A.[ J. Org. Chem., 63 (24), 8837-8842, 1998]; Miura, T. et a/, [Bioorg. Med. Chem., 6, 1481-1498, 1998]; Shin, S. et al., [Tetrahedron asymmetry, 11 , 3293-3301 , 2000]; WO 2002012185

A stereoselective synthesis of enantiomerically pure D-fftreo-PDMP has also been described by Shin, S. et a/., Tetrahedron asymmetry, 11 , 3293- 3301 , 2000 and WO 2002012185 the key step is the regioselective cleavage by nitrogen nucleophiles, as morpholiπe, of the C(3)-N-bond of non-activated enantiomerically pure aziridine-2-methanols.

D-tfireo-PDMP 81 % i) TMS-I, CH₃CN ii) a) morpholine b) HCI in) Pd(OH)₂ H₂ AcOH MeOH, 40^φC ιv)10% NaOH decanoyl chloride 81%

On the other hand, the synthesis of enantiomerically pure (1S,2S)-1-phenyl-2- decanoylamino-3-morpholino-1-propanol (L-fhreo-PDMP) from L-serine has also been described by Mitchell, Scott A., J. Org. Chem., 63 (24), 8837-8842, 1998.

i) IBU₅AI₂H PhMgSr L-threo-POMP Docket 18174 PCT(AP)

Other known methods to obtain L-threo-PDMP are described by Miura, T et al, Bioorg Med Chem , 6, 1481-1498, 1998 and in JP-A-9-216858 Synthesis of (1 S.2S)- threo- and (1R,2S)-e/ytfiro-1-phenyl-2- palmιtoylamιno-3-N-morpholιno-i-propanol (PPMP) were described starting from Garner aldehyde of L-seπne, by Nishida, A , Synlett, 4, 389-390, 1998

L-fbreoPPMP 0-erythro-PPMP

D-threo-1 -phenyl-Σ-palmitoylamino-S-pyrrolidino-i -propanol (P4 or PPPP) analogues were first obtained by a Mannich reaction as described Abe, A et al . J Biochem , 111, 191-196, 1992 or US 5916911 and WO 2001004108

OH

Preparation of D-ftreo-4'-hydroxy-P4 was described by Lee, L ef a/ , J Biol Chem , 274, 21 , 14662-14669, 1999 In addition, a series of dioxane substitutions was designed and tested These included 3',4'- methylenedιoxyphenyl-3',4'- ethylenedioxyphenyl-, and 3',4'- tπmethylenedioxyphenyl-substituted homologues

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Synthesis of eπantiomerically pure D-fr)reo-1-phenyl-2- benzyloxycarbonylamino-3-pyrrolidιno-1-propanol (PBPP) and D-threo-P4 and its analogues from N-benzyloxycarbonyl-D-serine, was described by Jimbo M. et al, J. Biochem., 127(3), 485-91, 2000 and EP 782992 (Seikagaku Kogyo Co.).

D-(ftreo-PBPP

Novel prodrugs of P4 derivatives were described in US 20020198240 and WO 2002062777

Synthesis of enantiomerically pure of D-fftreo-ethylenedioxy-P4 and D- fftreo-p-methoxy-P4 were described by Husain A. and Ganem B., Tetrahedron Lett, 43, 8621-8623, 2002. The key step is a highly syn-selective additions of aryl Grignard reagents to Garner aldehyde.

Docket 18174 PCT (AP)

Garner Aldehyde

Diastereoselective synthesis of P4 analogues were described in US 03/0153768 and WO 2003045928 (Genzyme Corp ), Oxazolmes I [R1 = (un)substituted aryl, R², R³ = H, (un)substιtuted aliphatic, NR²R³ = heterocyclic] are prepared as intermediates for P4 glucosyltransferase inhibitors from R¹CHO and R²R³NCOCH₂CN Thus, methyl isocyanoacetate CNCH₂CO₂Me was treated with pyrrolidine and the amide was treated with 1,4-benzodιoxane-6-carboxaldehyde, followed by hydrolysis of the oxazoline using HCI in methanol, reduction of the keto group of amide Il using LiAIH₄, and acylation with palmitoyl chloride to give D,l_-frireo-ethylenedιoxy-P4 III

D L-threo 3 4 -ethylenedioxy P4

Synthesis of enantiopure P4 analogues were described in WO 2003008399 (Genzyme Corp )

P4 derivatives, such as I [R¹, R⁵ = un(substιtuted) aromatic, R², R³ = H, un(substιtuted) aliphatic, NR²R³ = (un)substιtuted non-aromatic heterocyclic ring, R⁴ = O, H₂], have been prepared Thus, D-tf?reo-ethylenedιoxy-P4 was

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prepared via a multistep synthetic sequence starting from S-(+)-Ph glycinol, pheπyl-α-bromoacetate, 1 ^-benzodioxan-β-carboxaldehyde, pyrrolidine and palmitoyl chloride

D threo-3 4 -ethylenedιoxy-P4

New D-tfireo-P4 analogues that bear ether substituents on the aromatic ring have been recently synthesized from D-seπne and found to suppress neuπte extension in an embryonic insect cell line as described by Slavish , J P et al , Bιoorg Med Chem Lett , 14, 1487-1490, 2004 Further references which serve as background to the present invention are United States Patent Nos 5,945,442, 5,952,370, 6,030,995 and 6,051 ,598, Kurosawa et al, Journal of Labelled Compounds & Radiopharmaceuticals (1996), 38(3), 285-97, Published PCT application WO 01/38228, and Kastron et al Latvijas PSR Zinatnu Akademijas Vestis, Kimijas Seπja (1965) (4), 474-7

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SUMMARY OF THE INVENTION

The present invention is directed to methods of treating a patient suffering from one or more types of cognitive disorders using compounds of Formula 1 :

Formula 2 where Ri is H or alkyl of 1 to 6 carbons,

R₂ is H, alkyl of 1 to 6 carbons or the Ri and R₂ groups together with the nitrogen form a saturated or unsaturated 4, 5, 6 or 7 membered ring that optionally includes one or two heteroatoms independently selected from N, O and S, said 4, 5, 6 or 7 membered ring optionally being substituted with a halogen, COOH, CH₂OH, OH, B(OH)₂, cyano or with an alkyl group having 1 to 6 alkyl groups, R₃ is independently selected from H, alkyl of 1 to 20 carbons, aryl or heteroaryl, aryl-alkyl or heteroaryl-alkyl where the alkyl moiety is has 1 to 4 carbons, cycloalkyl of 3 to 6 carbons, said aryl or heteroaryl groups being optionally substituted with 1 to 3 groups independently selected from the group consisting of halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons, or

R₃ is CO-R₇ or CO-O-R₇ where R₇ is H, alkyl of 1 to 1 to 20 carbons, benzyl, alkyl of 1 to 20 carbons substituted with and NH₂ group, with a NHCOOalkyl or with an NH-COalkyl group where the alkyl group has one to 6 carbons, or R₇ is aryl, heteroaryl, aryl-alkyl or heteroaryl-alkyl where the alkyl moiety is branched or unbranched and has 1 to 4 carbons, said aryl or heteroaryl 36

Docket 18174 PCT (AP)

groups being optionally substituted with 1 to 3 groups independently selected from the group consisting of halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons,

R_A IS H, alkyl of 1 to 6 carbons or CO-R₈ where R₈ is alkyl of 1 to 6 carbons, the wavy lines represent bonds connected to carbons having R or S configuration, and

Rio is selected from the groups of formulas (i) and (ii)

R₅

¹W

(I) (II) where the * indicates the carbon atom to which the remaining moiety of the molecule is attached,

R₅ and Re independently are H, alkyl of 1 to 6 carbons, halogen, alkoxy of 1 to 6 carbons or the R₅ and He groups together with the atoms to which they are attached jointly form a carbocyclic or a heterocyclic ring, the carbocyclic ring having 5 or 6 atoms in the ring, the heterocyclic ring having 5 or 6 atoms in the ring and 1 to 3 heteroatoms independently selected from N, O and S, and said carbocyclic or heterocyclic ring jointly formed by R₅ and R₆ being optionally substituted with 1 to 6 R₉ groups where Rg is independently selected from halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons, with the proviso that when Ri₀ has formula (11) then Formula 1 does not include compounds where R₄ is hydrogen and Ri and R₂ jointly with the nitrogen form a morpholin or a pyrrolidin ring and where R₅ and R_$ both are H or one of R₅ and R₆ is OCH₃ and the other is H, and the present invention is also directed to all pharmaceutically acceptable salts of said compounds Docket 18174 PCT (AP)

Any of the compounds described here may be used to treat a patient suffering from a cognitive disorder, such as an agnosia, an amnesia, an aphasia, an apraxia, a delirium, a dementia, and a learning disorder DETAILED DESCRIPTION OF THE INVENTION A general description of the compounds of the invention is provided in the Summary Section of the present application for patent Most compounds of the invention contain one or more asymmetric centers, such that the compounds may exist in enantiomeric as well as in diastereomeπc forms In fact, most of the compounds of the present invention have two asymmetric carbons adjacent to one another and therefore can exist in erythro or threo form, with each of these two forms having dextrorotatory (D) or levorotary (L) enantiomers Although the fftreo form is generally preferred in accordance with the present invention, unless it is specifically noted otherwise, the scope of the present invention includes all enantiomers, diastereomers and diastereomeric and racemic mixtures In light of the foregoing, it should be clearly understood that the designation "DL" or "(+/-)" or " (±)" in this application includes the pure dextrorotatory enantiomer, the pure levorotatory enantiomer and all racemic mixtures, including mixtures where the two enantiomers are present in equal or in unequal proportions Moreover, for simplicity sake in many of the structural formulas, such as in the example below, only one of the enantiomers is actually shown but when the designation "DL" (or "(+/-)" or "(±)") appears it also includes the enantiomeric form (mirror image) of the structure actually shown in the formula For Example

OH

N_x^ RH₂ L-/ DL-fhreo or ±-threo Docket 18174 PCT (AP)

Thus, in the example above, only one enantiomer is shown, but because the designation "DL" (or "(+/-)" or "(±)") appears below the formula, its optical isomer

and all racemic mixtures of the two optical isomers are also included In the case of some compounds of the present invention one enantiomer of the threo, and in some cases of the erythro, is significantly more active than the other enantiomer of the same pair Some of the compounds which may be used in the method of the present invention may contain three or more asymmetric centers

Keeping the foregoing examples in mind a person of ordinary skill in the art should readily understand the scope of each described example, although in a broad sense all isomers, enantiomers and racemic mixtures are within the scope of the invention

The term "alkyl" in the general description and definition of the compounds includes straight chain as well as branch-chained alkyl groups

Generally speaking the compounds of the invention may form salts with pharmaceutically acceptable acids or bases, and such pharmaceutically acceptable salts of the compounds of Formula 1 are also within the scope of the invention

Referring now to the novel compounds of Formula 1 the Re and Re groups preferably both are independently selected from H, alkyl, alkoxy and still more preferably are H In the preferred compounds the R₃ groups are preferably both H, or one of the R₃ groups is H and the other is an acyl group or an arylalkylcarbamoyl group The R₄ group is preferably H (but see the "proviso" in the Summary section) or alkanoyl, and the Ri and R₂ groups preferably are pyrrolidino or morpholino Docket 18174 PCT (AP)

BIOLOGICAL ACTIVITY, MODES OF ADMINISTRATION The compounds described here may be used to treat a patient suffering from one or more types of cognitive disorder, such as an agnosia, an amnesia, an aphasia, an apraxia, a delirium, a dementia, and a learning disorder To "treat," as used here, means to deal with medically It includes, for example, administering a compound of the invention to prevent the onset of a cognitive disorder, to alleviate its severity, and to prevent its reoccurrence

The term "cognitive disorder," as used here, means any condition characterized by a deficit in mental activities associated with thinking, learning, or memory Examples of such disorders include agnosias, amnesias, aphasias, apraxias, deliriums, dementias, and learning disorders In some cases, the cause of a cognitive disorder may be unknown or uncertain In other cases, the cognitive disorder may be associated with (that is, be caused by or occur in the presence of) other conditions characterized by damage to or loss of neurons or other structures involved in the transmission of signals between neurons Hence, cognitive disorders may be associated with neurodegenerative diseases such as Alzheimer's disease, corticobasal degeneration, Creutzfeldt-Jacob disease, frontotemporal lobar degeneration, Huntington disease, multiple sclerosis, normal pressure hydrocephalus, organic chronic brain syndrome, Parkinson's disease, Pick disease, progressive supranuclear palsy, or senile dementia (Alzheimer type), it may be associated with truama to the brain, such as that caused by chronic subdural hematoma, concussion, intracerebral hemorrhage, or with other injury to the brain such as that cause by infection (e g , encephalitis, meningitis, septicemia) or drug intoxication or abuse

Cognitive disorders may also be associated with other conditions which impair normal functioning of the central nervous system, including psychiatric disorders such as anxiety disorders, dissociative disorders, mood disorders, schizophrenia, and somatoform and factitious disorders, it may also be associated with conditions of the peripheral nervous system, such as chronic pain Docket 18174 PCT (AP)

The compounds described here may be used to treat agnosias, amnesias, aphasias, apraxias, deliriums, dementias, learning disorders and other cognitive disorders regardless of whether their cause is known or not Examples of dementias which may be treated with the methods of the invention include AIDS dementia complex, Binswanger's disease, dementia with Lewy Bodies, frontotemporal dementia, multi-mfarct dementia, Pick's disease, semantic dementia, senile dementia, and vascular dementia

Examples of learning disorders which may be treated with the methods of the invention include Asperger's syndrome, attention deficit disorder, attention deficit hyperactivity disorder, autism, childhood disintegrative disorder, and Rett syndrome

Examples of aphasia which may be treated with the methods of the invention include progressive non-fluent aphasia

The compounds described here may also be used to treat patient having deficits in mental activities that are mild or that otherwise do not significantly interfere with daily life Mild cognitive impairment is an example of such a condition a patient with mild cognitive impairment displays symptoms of dementia (e g , difficulties with language or memory) but the severity of these symptoms is such that a diagnosis of dementia may not be appropriate The compounds described here may be used to treat mild cognitive impairment and other, similarly less severe forms of cognitive disorders Examples of Compounds of the Invention

Table 1 , below, lists compounds which may be used in the method of the invention

TABLE 1

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Modes of Administration:

Compounds useful in the methods of the invention may be administered at pharmaceutically effective dosages Such dosages are normally the minimum dose necessary to achieve the desired therapeutic effect, in the treatment of chromic pain, this amount would be roughly that necessary to reduce the discomfort caused by the pain to tolerable levels For human adults such doses generally will be in the range of O 1-5,000 mg/day, more preferably in the range of 1 to 3,000 mg/day, 10 mg to 500 mg/day, 500 to 1,000 mg/day, 1,000 to 1,500 mg/day, 1,500 to 2,000 mg/day, 2,000 to 2,500 mg/day, or 2,500 to 3,000 mg/day However, the actual amount of the compound to be administered in any given case will be Docket 18174 PCT (AP)

determined by a physician taking into account the relevant circumstances, such as the severity of the pain, the age and weight of the patient, the patient's general physical condition, the cause of the pain, and the route of administration The compounds are useful in the treatment of pain in a mammal, particularly a human being Preferably, the patient will be given the compound orally in any acceptable form, such as a tablet, liquid, capsule, powder and the like However, other routes may be desirable or necessary, particularly if the patient suffers from nausea Such other routes may include, without exception, transdermal, intrapeπtonial, parenteral, subcutaneous, intranasal, intrathecal, intramuscular, intravenous and intrarectal modes of delivery Compositions useful in the method of the invention may further include an excipient Such an excipient may be a earner or a diluent, this is usually mixed with the active compound, or permitted to dilute or enclose the active compound If a diluent, the carrier may be solid, semi-solid, or liquid material that acts as an excipient or vehicle for the active compound The formulations may also include wetting agents, emulsifying agents, preserving agents, sweetening agents, and/or flavoring agents If used as in an ophthalmic or infusion format, the formulation will usually contain one or more salt to influence the osmotic pressure of the formulation

SYNTHETIC METHODS FOR OBTAINING THE COMPOUNDS OF THE

INVENTION, EXPERIMENTAL

The compound of the invention can be synthesized by utilizing the synthetic methods described in the experimental below, or such modifications of the below described experimental methods which will become readily apparent to those skilled in the art in light of the present disclosure

GENERAL

¹H NMR spectra were recorded at ambient temperature with an Avance 300 (Bruker) spectrometer The compounds were analyzed by reverse phase high performance liquid chromatography (HPLC) using a Waters Autopurification Docket 18174 PCT (AP)

System equipped with a Waters 2525 Pump a Waters 2696 photodiode array detector, and a XTerra column (Part No 186000482, 5 μm, C18, 4 5 * 50 mm)

The HPLC method used was a gradient of 5 % solvent B to 100 % in 7 mm Solvent A was H₂O with 0 05 % TFA and solvent B was CH₃CN with 0 05 % TFA (Method A)

Melting points were measured with a Buchi B-545 melting point apparatus and were uncorrected To isolate reaction products the solvent were removed by evaporation using a vacuum rotatory evaporator the water bath temperature not exceeding 40 °C

GENERAL SYNTHETIC ROUTES

The compound of the invention can be synthesized by utilizing the synthetic methods described in a general sense immediately below and in more detail in the experimental section of the present application, or by such modifications of the below described general and experimental methods which will become readily apparent to those skilled in the art in light of the present disclosure

A general synthetic route to the compound of the present invention which are substituted "1-hydroxyl-propyl amines" may lead through the synthesis of the corresponding substituted "3-hydroxyl-propyl amide ' compounds, followed by reduction of the carbonyl group of the "carboxylic acid amide" moiety with a reducing agent such as lithium aluminum hydride, or like reducing agent

OH O OH

NH₂ R₂ LiAIH₄ ^NH2 R₂

General Structure 1 General Structure 2

General Synthetic Scheme A T7US2008/054936

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This reaction is illustrated in General Synthetic Scheme A, where, generally speaking, the variables have the meaning described in the Summary Section of the present application for patent A person of ordinary skill in the art of organic synthesis will nevertheless readily understand that depending on the nature of the substituents designated R-i, R₂ and R₁₀ certain groups may need to be protected for the performance of the reduction step

The substituted "3-hydroxyl-propyl amide" compounds can, generally speaking, be synthesized as described below in the following General Reaction Scheme 1 and General Reaction Scheme 2

methylisocyanoacetate "amine" 2-Isocyano-l-(subst. -arnιno)ethanoιie

R* -CHO strong base "aldehyde" (KOH)

(W-ythreo fraπs-oxazoline General Structure 1

General Reaction Scheme 1

Thus, in accordance with General Scheme 1 , methyl isocyanoacetate (or ethyl isocyanoacetate available commercially) is reacted with an "amine" Docket 18174 PCT (AP)

which includes the R₁ and R₂ groups to provide the 2-ιsocyanoacetιc acid amide derivative shown in General Reaction Scheme 1 Typical examples for the amines used in the reaction are pyrrolidine, pipeπdine, azetidme, morpholme, 2,5-dιhydro-1H-pyrrole, dialkylamines such as diethylamine, 3- fluoro-, 3,3-dιfluoro or 3-hydroxy substituted pyrrolidines The 2- isocyanoacetic acid amide derivative is then reacted in methanol in the presence of base (such as KOH) with an "aldehyde" which includes the Rm group to provide a trans "oxazoline" with high d ^stereoselectivity (trans cis ratios generally > 97 3) as shown in General Reaction Scheme 1 The trans oxazoline is then treated in methanol with a strong acid, such as HCI, to open the ring and to provide the fhreo-3-substιtuted-3-hydroxy-2-amιno-propιonιc acid amide intermediates (with a threo erythro ratios generally > 97 3) as shown in General Reaction Scheme 1

Compounds of Formula 1 and or of General Structure 1 , where the ammo group of formula NHR₁R₂ is a weaker nucleophile, such as indolme, thiomorpholine and the like, can be made as illustrated in Reaction Scheme 2 for the synthesis of intermediate compounds (±)-fhreo-2-amιno-3-hydroxy-1- (ιndolιn-1-yl)-3-(pyrιdιn-4-yl)propan-1-one dihydrochloπde Compound 243 and (+Hhreo-2-amino-3-hydroxy-1-(thιazolιdιn-3-yl)-3-(pyπdιn-4-yl)propan-1- one dihydrochloride Compound 242

Docket 18174 PCT (AP)

Me (ert butyl

(±) thieo 30 % yield (±) erythro 2 % yield

(±) tftreo

Compound 243

(±) fhΛBO

Compound 242

a) KOH MeOH b) lndoline EDCI TEA HOBT CH₂CI₂ c) HCI (1M) in MeOH d) i Silica Gel Chromatography n HCI (O 1M) in /-PrOH e) BOC₂O NaOH Dioxane f) Thiazolidine EDCI TEA HOBT CH₂CI₂ g) Silica Gel Chromatography g) HCI (1M) in MeOH

Reaction Scheme 2

In Reaction Scheme 2 EDCI stands for 1-(3-dimethylaminopropyl)- ethylcarbodnmide hydrochloride, HOBT stands for 1-hydroxybenzotπazole, BOC₂O stands for di-f-butyl-dicarbonate and TEiA stands for tπethylamine US2008/054936

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Compounds 242 and 243 can be reduced, as illustrated in General Synthetic Scheme A to provide compounds of the invention

Another general synthetic route may follow in general terms the synthesis of Compound 1 , Compound 2 and Compound 3, specifically described in detail in the experimental section below, modified with such modifications which in light of the present disclosure will become readily apparent to a person of ordinary skill in the art lsomerically pure and/or enantiomerically pure compounds and further derivatives of the 3-substιtuted-3-hydroxy-2-amιno-propιonιc acid amide intermediates or of the substituted 1 -hydroxy propylamines of the invention are obtained by separation techniques and reactions which per se, are well known to the synthetic chemist Some of the typical separation techniques and reactions are generally described below

Separation of threo and erythro isomers, when both are formed in the reactions leading to the compounds of the invention, can typically be performed by chromatographic methods The chromatographic separation may occur the level of the substituted 3-hydroxyl-propιonιc acid amide intermediate compounds or at the level of the substituted 1-hydroxyl propyl amine compounds of the invention The more abundantly formed threo isomers can also be converted into the erythro isomers by oxidizing to the ketone level the hydroxyl group in the 3 position of the propanoic acid moiety and subsequently reducing the resulting ketone to the hydroxyl level in the intermediate 3-substιtuted-3-hydroxy-2- amino-propionic acid amide compounds or in the compounds of the invention Separation of enantiomeric mixtures can be performed on Chiralpack columns which are well known in the art

The amino function in the 2-posιtιon of the propyl amine moiety is, generally speaking, more reactive towards acylation and carbamoylation than the hydroxyl group in the 1 position Therefore, acylated derivatives of the 2- amino function can be prepared by using acyl chlorides such as acetyl chloride and hexanoyl chloride Or the 1 -hydroxy and 2-amιno groups of the T/US2008/054936

DockeM8174 PCT (AP)

compounds of the invention can be acylated in the same reaction Carbamate derivatives of the 2-amιno function can be obtained by using chloroformates, such as benzylchloroformate A tertiary butyl carbamoyl function or benzyl- carbamoyl function can also serve as a removable protecting group of the 2- amino function

Alkylation of the 2-amιno function can be performed by condensing the compound bearing the 2-NHa group with an aldehyde to obtain a Schiff base intermediate which can be reduced, without isolation, to provide the N-alkyl, arylalkyl or heteroaryl-alkyl compounds of the invention

DETAILED DESCRIPTION OF THE SYNTHESIS OF PREFERRED COMPOUNDS (EXPERIMENTAL)

Preparation of D-tf?reo-2-amιno-3-morpholιno-1-phenylpropan-1-ol dihydrochloride Compound 4

(R)-Methyl 1-((S)-1-phenylethyl)azιrιdιne-2-carboxylate EBE 06044B

To solution of methyl 2,3-dιbromopropionate (25 mL, 198 mmol) in toluene at 5 °C was added triethylamine (55 mL, 039 mmol) in toluene (100 mL) After stirring for 5 mm (S)-(I )-ρhenethylamιne (25 mL, 198 mmol) in toluene (100 mL) was added dropwise The suspension was refluxed for 3 h and allowed to cool down, filtered and the volatiles were evaporated under reduced pressure to give a residue that was purified by column chromatography (950 g of silica gel) with a gradient of 0-20 % EtOAc in cyclohexane to yield to (S)-methyl 1-((S)-1-phenylethyl)azιπdιne-2-carboxylate EBE 06044A as a yellow oil (17 31 g, 43 % yield) and (R)-methyl 1-((S)-1- phenylethyl)azιrιdιne-2-carboxylate EBE 06044B as a yellow oil (15 14 g, 37 % yield)

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EBE 06044B

MW 205 3, Yield EBE 06044B 37 %, Yellow Oil Yield EBE 06044A 43 % Yellow Oil

5 R_f EBE 06044A = 0 5, R, EBE 06044B = 035 (EtOAc cyclohexane = 2575)

¹H-NMR (CDCI₃,) EBE 06044A 1 47 (d, 3H, J = 66 Hz, CH₃), 1 60 (d, 1H, J = 64 Hz, CH), 2 13 (d, 1H, J = 2 6 Hz), 2 21 (dd, 1H, J = 32 Hz, J = 64Hz), 2 54 (q, 1 H, J = 6 6 Hz), 375 (s, 3H, OCH₃) 7 23-7 40 (m, 5H, ArH)

10 ¹H-NMR (CDCI₃, δ) EBE 06044B 1 46 (d, 3H, J = 6 6 Hz, CH₃), 1 79 (d, 1H, J = 6 6 Hz, CH), 2 08 (d, 1H, J = 3 11Hz, 66 Hz), 2 34 (dd, 1H₁ J = 3 1 Hz, J = 1 0 Hz), 2 56 (q, 1H, J = 6 6 Hz), 3 67 (s, 3H, OCH₃) 7 24-7 36 (m, 5H, ArH) 1³C-NMR (CDCI₃, δ) EBE 06044B 23 5, 350, 36 9, 52 2, 698, 126 5, 1272, 1285, 1436, 171 1

15 HPLC Method A, detection at 254 nm, EBE 06044B RT = 6 11 mm, peak area 92 9 %

((R)-I -((S)-1-Phenylethyl)azιπdιn-2-yl)methanol EBE 06046

A 250 mL round bottom flask was charged with anhydrous THF

20 (10OmL) and LiAIH₄ (2 77 g, 73 1 mmol) While the suspension is stirred at 0 0C, a solution of (S)-methyl 1-((S)-1-phenylethyl)azιrιdιne-2-carboxylate EBE 06044B (10 0 g, 48 7 mmol) in THF (50 mL) was added dropwise over 20 mm The dropping funnel was washed with THF (2 x 3 mL) and allowed to react 20 mm at 0⁰C Maintaining the reaction mixture at O⁰C, a solution of KOH (10 %,

25 20 mL) was added dropwise for 20 mm (caution the reaction is exothermic) The mixture was stirred for 0 5 h at 25 ⁰C and the white precipitate removed by filtration through a celite pad that was washed with diethyl ether (30 mL) 008/054936

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The combined organic filtrates were washed with NaH₂PO₄ and the aqueous layer was extracted with Et₂O (3 * 30 mL) The combined organic phase were dried with Na₂SO₄ and concentrated to give ((R)-1-((S)-1-phenylethyl)azιπdιn- 2-yl)methanol EBE 06046 as a white solid (104 g, 90 % yield)

OH

L

N

EBE 06046

MW 177 2, Yield 90 %, White Solid, Mp (⁰C) 377 1H-NMR (CDCI₃, δ) 1 43 (d, 3H, J = 6 6 Hz, CH₃), 1 49 (d, 1 H, J = 6 5 Hz, CH), 1 65-1 71 (m, 1H, CH), 1 92 (d, 1H, J = 3 5 Hz, NCH), 2 26 (s, 1H, OH), 10 2 53 (q, 1H, J = 6 6 Hz, NCH), 3 32-3 37 (m, 1H, OCH₂), 3 56 (m, 1H₁ OCH₂), 7 23-7 35 (m, 5H, ArH) 1³C-NMR (CDCI₃, δ) 22 9, 31 4, 39 3, 62 5, 694, 126 6, 127 3, 128 6, 144 5

(R)-1-((S)-1-Phenylethyl)azmdιne-2-carbaldehyde EBE 06048 15 A three neck, 250 mL round bottom flask was equipped with a low temperature thermometer and two (2) equalizing dropping funnels One of these was connected to a nitrogen line and charged with a solution of ((R)-I- ((S)-I -phenylethyl)azιrιdin-2-yl)methanol EBE 06046 (7 0 g, 39 5 mmol) in CH₂CI₂ (75 mL), the other was charged with a solution of DMSO (9 25 g, 20 1185 mmol) in CH₂CI₂ (11 mL) To a solution of oxalyl chloride (7 5 g, 593 mmol) in CH₂CI₂ (90 mL) under N₂ at -78 "C, the DMSO solution was added dropwise during 20 mm and stirred for 20 mm EBE 06046 (7 0 g, 39 5 mmol) in CH₂CI₂ (75 mL) was added dropwise over 50 mm then the dropping funnel was charged with DIEA (42 6 mL, 237 mmol) in CH₂CI₂ (10 mL) and the 25 reaction mixture was stirred for 30 min at ~45°C The DIEA solution was added over 5 mm with the reaction mixture at -78 ⁰C and the reaction was allowed to warm to room temperature The reaction mixture was washed with 54936

Docket 18174 PCT (AP)

H₂O (3 * 50 mL), dried over MgSO₄, filtered, evaporated The crude product obtained was purified by column chromatography on silica with a gradient of 0-20 % [v/v] EtOAc in cyclohexane to give (R)-1-((S)-1-phenylethyl)azιrιdιne- 2-carbaldehyde EBE 06048 as a yellow oil (5 59 g, 81 % yield)

EBE 06048

MW 1752, Yield 81 %, Yellow Oil

R_f EBE 06048 0 3 (EtOAc cyclohexane = 20 80)

¹H-NMR (CDCI₃, δ) 1 47 ( d, 3H, J = 6 6 Hz, CH₃), 1 94 (d, 1H, J = 6 7 Hz, NCH₂), 2 08 (dt, J = 2 9 Hz, J = 6 4 Hz, NCH), 2 37 (d, 1H, J = 2 6 Hz, NCH₂),

2 61 ( q, 1H, J = 6 6 Hz, NCH), 7 20-7 38 (m, 5H₁ ArH) 892 (d, 1H₁ J = 6 2

Hz)

¹³C-NMR (CDCI₃, δ) 22 7, 32 1 , 43 2, 68 1 , 125 5, 126 5, 127 6, 142 4,

1987

(R)-Phenyl((R)-1-((S)-1-phenylethyl)azιrιdιn-2-yl)methanol EBE 06066

To a solution of bromobenzene (493 g, 31 4 mmol) in THF 125 mL under nitrogen at - 78 °C was added f-BuLi (1 7 M in pentane, 50 mL) The mixture was stirred for 0 5 h at room temperature The mixture was cooled down to -78 ⁰C and a solution of (R)-1-((S)-1-phenylethyl)azιπdιne-2- carbaldehyde EBE 06048 (2 5 g, 14 3 mmol) in THF (16 7 mL) at -78 ⁰C was added dropwise The reaction mixture was treated with H₂O (20 mL), the organic layer was separated and the aqueous phase was extracted with

EtOAc The combined organic layers were dried over MgSO₄, filtered and concentrated in vacuo to give a residue that was purified by column chromatography using a gradient of 0-20 % [v/v] EtOAc in cyclohexane to give 54936

Docket 18174 PCT (AP)

(R)-phenyl((R)-1-((S)-1-phenylethyl)azmdin-2-yl)methanol EBE 06066 (3 13 g, 86 % yield)

5 EBE 06066

MW 253 3, Yield 86 % R_f = 0 3 (EtOAc cyclohexane = 20 80)

¹H-NMR ( CDCI₃, δ) 1 47 (d, 3H, J - 66 Hz, CH₃), 1 57 (d, 1H, J = 6 5 Hz, CH), 1 79 (dt, 1 H, J = 3 5 Hz, J = 8 7 Hz, CH), 2 04 (d, 1H, J = 3 5 Hz, OCH), 10 2 35 (bs, 1H, OH), 2 53 (q, 1H, J = 6 5 Hz, CH), 4 23 (d, 1H, J = 5 7Hz, OCH), 7 07-7 13 (m, 2H, ArH), 7 16-7 20 (m, 3H, ArH), 7 24-7 34 (m, 5H, ArH) 1³C-NMR (CDCI₃, δ) 224, 32 O, 44 6, 694, 74 1, 125 8(2xC), 126 9 (2xC), 127 3, 1276, 128 2 (2xC), 1287 (2xC), 142 O, 1442 [α]²² _D = - 71 53 (c = O 59, CHCI₃)

15

D-f/7reo-2-((S)-1 -Phenylethylamιno)-3-morpholιno-1 -phenylpropan-1 -ol dihvdrochloride Compound 5

To a solution of (R)-phenyl((R)-1-((S)-1-phenylethyl)azιπdιn-2- yl)methanol EBE 06066 (1 5 g, 5 92 mmol) in CH₃CN (19 mL) at RT was

20 added iodotrimethylsilane (3 55 g, 17 8 mmol) The solution was stirred for 2 h and morpholine (1 032 g, 11 84 mmol) was added After 2 h at reflux, the reaction mixture was treated with HCI (1M) to reach pH = 1 and stirred for 10 mm After a slow addition of NaHCO₃ to reach pH = 9, the product was extracted with EtOAc, dried over Na₂SO₄, filtered to give after evaporation a

25 crude brown oil that was purified by column chromatography using a gradient of 0-20% [v/v] MeOH in EtOAc to give D-f/7reo-2-((S)-1-phenylethylamιno)-3- morpholιno-1 -phenylpropan-1 -ol EBE 06068A (0 831 g, 42 %) as a pale 008/054936

Docket 18174 PCT (AP)

brown solid To a solution of D-tf?reo-2-((S)-1-phenylethylamιno)-3- morpholιno-1-phenylpropan-1-ol EBE 06068A (0 100 g, 0294 mmol) in ethanol (1 mL) was added a solution of HCI (08 M, 0 816 mL) in EtOH Evaporation of the volatiles afforded to D-t/ireo-2-((S)-1-phenylethylamιno)-3- morpholιno-1-phenylpropan-1-ol dihydrochloride Compound 5 as white solid (0 125 g, 100 %)

Compound 5 MW 412 37 Yield 42 %, White Solid, Mp (⁰C) 1572 (dec) Rf 0 3 (MeOH EtOAc = 20 80) EBE 06068A

¹H-NMR (CD₃OD, δ) 1 19 (t, 2H, J = 7 0 Hz, NCH₂), 1 71 (d, 3H, J = 6 8 Hz, CH₃), 345 (m, 2H, J = 7 1 Hz, NCH₂), 3 62 (q, 2H, J = 7 1 Hz, N-CH₂), 3 97 (t, 4H, J = 4 5 Hz, OCH₂), 4 06 (m, 1H, CH-N), 4 75 (q, 1H, J = 6 8 Hz, CH-N), 521 (d, 1H, J = 5 1 Hz, CH-O), 744-756 (m, 1OH, ArH) MS-ESI m/z (% rel lnt ) 341 1 ([MHf, 20)

¹³C-NMR (CD₃OD₁ δ) 244, 54 5 (2xC), 55 5, 559, 600, 67 0 (2xC), 756, 126 3 (2xC), 126 5 (2xC), 127 0, 127 1 , 128 1 (2xC), 128 5 (2xC), 142 2, 145 3 HPLC Method A, detection at 254 nm, Compound 5 RT = 441 mm, peak area 99 %

Threo-2-Amιno-3-morpholιno-1-phenylpropan-1-ol dihvdrochloride Compound 4

To a solution of D-tfireσ-2-((S)-1-phenylethylamιno)-3-morpholιno-1- phenylpropan-1-ol EBE 06068A (0400 g, 1 17 mmol) in MeOH (6 mL) at RT was added acetic acid (0 133 mL, 2 35 mmol) The reaction vessel was flushed with nitrogen and Pd(OH)₂ (25 % weight, 0 150 g) was added The 8 054936

Docket 18174 PCT (AP)

nitrogen atmosphere was exchanged with hydrogen using three cycle of vacuum and hydrogen addition using a balloon of hydrogen After stirring for 16 h under hydrogen the reaction mixture was filtrated through celite to give EBE 06070A the acetate salt of (2R)-amιno-3-morpholιn-4-yl-(1R)-phenyl- propan-1-ol (0 279 g, 98 % yield) To as solution of EBE 06070A the acetate salt of (2R)-amιno-3-morpholιn-4-yl-(1R)-phenyl-propan-1-ol (0 100 g, 0338 mmol) in ethanol (1 ml_) was added a solution of HCI (0 8 M, 0930 mL) in EtOH Evaporation of the volatiles afforded to D-fhreo-2-amino-3-morpholino- 1-phenylpropan-1-ol dihydrochloride Compound 4 (0 104 g, 100 % yield) as an off white solid (Adapted from Shin, S-H , Han, E Y , Park, C S , Lee, W K , Ha, H -J Tetrahedron Asymmetry, 2000, 11, 3293-3301)

Compound 4

MW 30923, Yield 99 %, Off White Solid, Mp (⁰C) 1834 1H-NMR (CD₃OD, δ) 3 30-3 77 (m, 6H, CH₂N), 3 92-4 05 (m, 4H, CH₂O),

4 05-4 16 (m, 1H₁ CH), 4 85-4 98 (m, 1H, CH), 7 35-7 60 (m, 5H, ArH)

¹³C-NMR (CD₃OD₁) 53 2, 58 3, 58 5 (2xC), 64 9 (2xC), 72 6, 128 0 (2xC),

130 2 (2xC), 140 3

MS-ESI m/z (% rel int ) 237 1 (100, [MH]⁺) HPLC lsocratic 10 % CH₃CN in H₂O (pH 10, [NH₄OH] = 5 mM), detection UV

254 nm, Compound 4 RT = 663 mm, peak area 97 3 %

[α]²² _D = - 10 7 (c = 1 00, MeOH)

Preparation of Benzyl L-f/?reo-1-hvdroxy-3-morpholιno-1-phenylpropan-2- ylcarbamate hydrochloride Compound 1

Benzyl (S)-3-hydroxy-1-oxo-1-phenylpropan-2-ylcarbamate TTA 0801 OB 36

Docket 18174 PCT (AP)

To a stirred solution of Z-L-Ser-OH (6 00 g, 25 08 mmol) in 32 mL of anhydrous THF at 0⁰C under nitrogen was added dropwise 1 M phenylmagnesium bromide in THF (32 mL, 200 mmol) (The symbol Z designates a benzylcarbamoyl group) The mixture was stirred 15 h at RT

5 under nitrogen A solution of 2 M HCI (100 mL) was slowly added at 0°C and the mixture was partitioned between ethyl acetate (750 mL) and acidic water The organic layer was washed with water (2 x 20 mL), 1 N aqueous sodium bicarbonate (2 x 20 mL), brine (2 x 20 mL) and dried over MgSO₄ After removing ethyl acetate by evaporation at 30-35 ⁰C, the crude product (450 g,

10 60 % yield) was cπstallized in a mixture of ethyl acetate hexane = 25 mL 20 mL to give benzyl (S)-3-hydroxy-1-oxo-1-phenylpropan-2-ylcarbamate TTA 0801 OB as a white solid (1 40 g, 20 % yield)

15 TTA 0801 OB

MW 299 32, Yield 20 %, White Solid, Mp (⁰C) 106 5

R_f 075 (CH₂CI₂ MeOH = 9 1)

¹H-NMR (CDCI₃, δ) 2 78 (s, 1H, OH), 3 85-393 (m, 1H, CH₂O), 4 00-4 09 20 (m, 1H, CH₂O), 5 14 (s, 2H, ArCH₂O), 540 (t, 1H, J = 3 3 Hz, CH), 6 17 (d

1 H, J = 64 Hz, NH), 7 35 (s, 5H, ArH), 749 (t, 2H, J = 7 60 Hz, ArH), 7 62 (t,

1 H, J = 7 1 Hz, ArH), 8 99 (t, 2H, J = 7 6 Hz, ArH)

¹³C-NMR (CDCI₃, δ) 58 3, 64 6, 67 3, 128 1, 128 3, 128 6, 128 7, 129 0,

134 1 , 136 0, 156 6, 196 6 25 MS-ESI m/z (% rel lnt ) 300 1 ([MH]⁺, 5), 256 1 (100)

HPLC Method A, detection UV 254 nm, TTA 08010B RT = 540 mm, peak area 98 5 %

Ia]²²D = - 5 8 (c = 1 00, MeOH) 36

Docket 18174 PCT (AP)

Benzyl L-tf?reo-1.S-dihydroxy-i-phenylpropan^-ylcarbamate TTA 08012

To a stirred solution of benzyl (S)-3-hydroxy-1-oxo-1-phenylpropan-2- ylcarbamate TTA 0801 OB (1 40 g, 4 70 mmol) in 28 mL of anhydrous THF at - 78 ⁰C under nitrogen was added slowly dropwise 1 M DIBAL-H in hexane (18 8 mL, 18 80 mmol) The mixture was stirred 2 h at -78 ⁰C then 1 5 h at RT A solution of 2 M HCI (35 mL) was slowly added at -20 ⁰C and the mixture was partitioned between ethyl acetate (750 mL) and acidic water The organic phase was washed with water (2x20 mL), brine (2x20 mL) and dried over MgSO₄ After removing ethyl acetate by evaporation at 30-35 ⁰C, the crude product was purified by column chromatography on silica (CH2CI2 MeOH = 982 to 97 3) to give benzyl L-f/7reo-1,3-dιhydroxy-1-phenylpropan-2- ylcarbamate TTA 08012 as a white solid (1 10 g, 78 % yield)

TTA 08012

MW 301 34, Yield 78 %, White Solid, Mp (⁰C) 102 5

Rf 0 30 (CH₂CI₂ MeOH = 95/5)

¹H-NMR (CDCI₃, δ) 3 08 (t, 1H, J = 5 0 Hz, OH), 3 59 (d, 1H, J = 3 1 Hz, OH), 3 64-3 78 (m, 2H, CH₂O), 3 80-3 89 (m, 1H, CH), 4 95 (s, 2H, ArCH₂O),

5 57 (d, 1H, J = 8 3 Hz, NH), 7 17-7 38 (m, 10H, ArH)

¹³C-NMR (CDCI₃, δ) 57 5, 63 6, 66 9, 73 8, 126 0 127 8, 127 9, 128 1,

128 5, 128 6, 136 2, 141 0, 156 9

MS-ESI m/z (% rel lnt ) 302 0 ([MH]⁺, 5), 132 0 (100) HPLC Method A, detection UV 254 nm, TTA 08012 RT = 5 00 mm, peak area

99 5 %

[α]²² _D = + 394 (c=1 00, MeOH) 2008/054936

Docket 18174 PCT (AP)

Benzyl threo-λ -hydroxy-3-morpholιno-1 -phenylpropan-2-ylcarbamate hydrochloride Compound 1

To a stirred solution of benzyl L-tfireo-1 ,3-dιhydroxy-1-phenylpropan-2- ylcarbamate TTA 08012 (1 0O g 3 30 mmol) in 13 mL of pyridine at -10 ⁰C

5 was added dropwise methanesulfonyl chloride (0 27 mL, 3 50 mmol) The mixture was stirred 6 h at 20 "C under nitrogen Pyridine was removed by evaporation at 30-35 ⁰C and the residue was partitioned between ethyl acetate (250 mL) and 0 1 N HCI (20 mL) The organic phase was washed with water (20 mL), brine (20 mL) dried over MgSO₄ and evaporated to give after

10 drying L-threo-λ -hydroxy-3-methanesulfonyl-1 -phenylpropan-2-ylcarbamate TTA 08014 (1 25 g , 65% yield)

To a stirred solution of crude benzyl L-tfireo-1-hydroxy-3-methanesulfonyl-1- phenylpropan-2-ylcarbamate TTA 08014 (1 25 g, 3 30 mmol) in 6 mL of DMF at RT was added morpholine (1 2 mL, 1320 mmol) The mixture was stirred

15 15 h at 50 ⁰C under nitrogen DMF was evaporated and the residue was partitioned between ethyl acetate (250 mL) and 1 N aqueous sodium bicarbonate (20 mL) The organic phase was washed with water (20 mL), brine (20 mL) and dried over MgSO₄ After evaporation the crude product was purified by column chromatography on silica (CH2CI2 MeOH = 98 2 to 97 3)

20 to give benzyl L-#7reo-1-hydroxy-3-morpholιno-1 -phenylpropan-2-ylcarbamate as an oil (380 rng, 31 % yield) The hydrochloride salt was obtained from 100 mg of the free base in diethylether at O⁰C using a solution 0 3 M HCI in diethylether The precipitate was filtered and dry to give benzyl L-f/?reo-1- hydroxy-3-morpholιno-1 -phenylpropan-2-ylcarbamate hydrochloride

25 Compound 1 as a white solid (70 mg, 65 % yield)

Compound 1 008/054936

Docket 18174 PCT (AP)

MW 406 90, Yield 20 %, White Solid, Mp (⁰C) 144 5 Rf 040 (CH₂CI₂ MeOH = 95 5)

¹H-NMR (CD₃OD, δ) 3 14-377 (m, 6H, CH₂N), 3 70-4 07 (m, 4H, CH₂O), 4 30-4 33 (m, 1H, CH), 4 90-5 06 (m, 3H, CH, ArCH₂O), 7 20-743 (m, 1OH, ArH)

¹³C-NMR (CD₃OD, δ) 51 2, 51 8, 53 2, 59 3, 63 2, 66 3, 72 5, 125 8, 127 2,

127 3, 127 5, 127 8, 127 9

MS-ESI m/z (% rel lnt ) 371 0 ([MH]⁺ 100)

HPLC Method A, detection UV 254 nm, Compound 1 RT = 4 40 mm, peak area 96 5 %

[α]²²D = +13 9 (c = 1 00, MeOH)

Preparation of f/ireo-2-amιno-3-morpholιno-1-phenylpropan-1-ol dihydrochloπde Compound 2

To a stirred solution of benzyl L-f/yreo-1-hydroxy-3-morpholino-1- phenylpropan-2-ylcarbamate (Compound 1 , 0 26 g, 0 70 mmol) in 20 mL of MeOH at RT was added Pd-C 10 % (140 mg) The mixture was satured with hydrogen and stirred for 24 h at RT under hydrogen atmosphere (balloon) The catalyst Pd-C 10% was removed by filtration on celite and the solution was evaporated The crude product was purified by column chromatography on silica (CH₂CI₂ MeOH NH₄OH = 79"20 1 to 75 20 5) to give L-fftreo-2-amιno- 3-morpholιno-1-phenylpropan-1-ol as an oil (100 mg, 60 % yield) The hydrochloride salt was obtained from 83 mg of the free base in diethylether at 0 ⁰C using 0 3 M HCI in diethylether After precipitation in diethylether, filtration and drying L-f/7/Bθ-2-amιno-3-morpholιno-1-phenylpropan-1-ol dihydrochloride Compound 2 was obtained as a white solid (80 mg, 74 % yield)

OH

2 ^HCI Compound 2 08/054936

Docket 18174 PCT (AP)

MW 309 23, Yield 44 %, White Solid, Mp (⁰C) 1664-170 9

R, 0 20 (CH₂CI₂ MeOH = 9 1)

¹H-NMR (CD₃OD, δ) 3 30-377 (m, 6H, CH₂N), 392-405 (m, 4H, CH₂O),

4 05-4 16 (m 1H, CH), 4 85-498 (m, 1H, CH), 7 35-760 (m, 5H, ArH) 1³C-NMR (CD₃OD, δ) 53 1, 54 9, 58 5, 648, 726, 1272, 128 0, 130 2, 140 3

MS-ESI m/z (% rel lnt ) 237 0 ([MH]⁺, 100)

HPLC Method A, detection UV 254 nm, Compound 2 RT = 0 90 mm, peak area 98 0 % [α]²² _D = +10 8 (c = 1 00, MeOH), free base [α]²² _D = - 6 1 (c = 0 25, CHCI₃) Preparation of benzyl L-fhrβo-1-acetoxy-3-morpholιno-1-phenylpropan-2- ylcarbamate hydrochloride Compound 3

Benzyl L-(hreo-1-acetoxy-3-morpholιno-1-phenylpropan-2-ylcarbamate hydrochloride Compound 3 To a stirred solution of benzyl L-fΛreo-1-hydroxy-3-morpholιno-1- phenylpropan-2-ylcarbamate hydrochloride (Compound 1 , 0 510 g, 1 25 mmol) in 30 mL of CHCI₃ at RT were added slowly tπethylamine (700 μL, 5 00 mmol) and acetyl chloride (145 μL, 200 mmol) The mixture was stirred 10 h at RT under nitrogen and partitioned between a mixture of ice-water (20 mL) and CH₂CI₂ (100 mL) The organic layer was washed with brine (20 mL) and dried over MgSO₄ After evaporation the crude product was purified by column chromatography on silica (CH₂CI₂ MeOH = 99 5 0 5 to 98 2) to give benzyl L- fr)reo-1-acetoxy-3-morpholιno-1-phenylpropan-2-ylcarbamate as an oil (0420 g, 81 % yield) The hydrochloride salt was obtained from 45 mg of the free base in diethylether at 0 ⁰C using a solution of 0 3 M HCI in diethylether The precipitate was filtered and dry to give benzyl L-fr?reo-1-acetoxy-3-morpholιno- 1-phenylpropan-2-ylcarbamate hydrochloride Compound 3 as a white solid (40 mg, 82 % yield) 054936

Docket 18174 PCT (AP)

Compound 3

MW 44894, Yield 66 %, White Solid, Mp (⁰C) 69 9

R_f 0 70 (CH₂CI₂ MeOH = 95 5) 1H-NMR (CD₃OD, δ) 2 10 (s, 3H, CH₃), 3 14-344 (m, 4H, CH₂N), 3 70-400

(m, 4H, CH₂O), 4 51-4 53 (m, 1H, CH), 490-5 13 (m, 2H, ArCH₂O), 5 89 (d,

1H, CH), 7 28-7 48 (m, 1OH, ArH)

¹³C-NMR (CD₃OD, δ) 20 8, 52 0, 52 6, 59 7, 646, 68 0, 76 5, 127 7, 129 0,

129 2, 129 5, 129 8, 137 9, 158 7, 171 3 MS-ESI m/z (% rel lnt ) 413 0 ([MH]⁺, 100)

HPLC Method A, detection UV 254 nm, Compound 3 RT = 470 min, peak area 98 5 %

Preparation of DL-f/?reo-2-(Decanamιdo)-1 -(4-methoxyphenyl)-3-(pyrrolιdιn-1 - vDpropyl decanoate Compound 10 2-lsocyano-1-(pyrrolιdιn-1-yl)ethanone BLE 04098

To stirred and cooled (0⁰C) methyl isocyanoacetate (96 % technical grade, 5 0 g, 47 8 mmol) was slowly added in 0 75 h pyrrolidine (6 5 mL, 78 mmol) The mixture was stirred for 1 5 h with continued cooling and then concentrated The resulting oil was co-evaporated twice from CH₂CI₂ hexane to remove residual pyrrolidine 2-lsocyano-1-(pyrrolιdιn-1-yl)ethanone BLE

04098 was obtained as a yellow solid (6 85 g, 98 % yield) and used in the next step without purification

BLE 04098 8 054936

Docket 18174 PCT (AP)

MW 138 17, Yield 98 %, yellow solid, Mp (X) = 739 1H-NMR (CDCI₃, δ) 1 81-2 08 (m, 4H, 2xCH₂), 3 35-345 (m, 2H, -NCH₂), 3 50-3 60 (m, 2H, -NCH₂J, 4 23 (s, 2H, CH₂CO) 7Vans-(4,5-dιhydro-5-(pyπdιn-3-yl)oxazol-4-yl)(pyrrolιdιn-1-yl)methanone BLE 0411OB frans-(4,5-Dιhydro-5-(4-methoxyphenyl)oxazol-4-yl)(pyrrolιdιn-1-yl)rnethanone SLA 07074

To a stirred and cooled (0°C) solution of potassium hydroxide (0 37 g, 657 mmol) in methanol (30 mL) was added a mixture of 4-methoxy- benzaldehyde (0 88 mL, 7 23 mmol) and 2-ιsocyano-1-(pyrrolιdιn-1- yl)ethanone BLE 04098 (1 0 g, 6 57 mmol) The solution was stirred 4 h with continued cooling and then concentrated The residue was partitioned between ethyl acetate and water The organic layer was combined with additional ethyl acetate extracts, washed with aqueous sodium chloride and dried over MgSO₄ Concentration afforded a crude product as a glassy solid Flash chromatography over silica (ethyl acetate) yielded to frans-(4,5-dιhydro- 5-(4-methoxyphenyl)oxazoi-4-yl)(pyrrolιdιn-1-yl)methanone SLA 07074 as a pale yellow solid (1 2 g, 90 5 %)

SLA 07074

MW 274 32, Yield 90 5 %, pale yellow solid, Mp (⁰C) 91 2

R_f 0 30 (EtOAc)

¹H-NMR (CDCI₃, δ) 1 75-2 08 (m, 4H, 2xCH₂), 340-3 58 (m, 3H, CH₂N),

3 52 (s, 3H, CH₃O), 388-3 98 (m, 1H, CH₂N), 4 59 (dd, 1H₁ J= 76 Hz₁ J = 2 2 Hz, CH-N), 606 (d, 1H, J = 76 Hz, CH-O), 6 90 (d, 2H, J = 87 Hz, ArH), 7 01 (d, 1 H, J = 2 2 Hz, CH=N), 7 25 (d, 2H, J = 87 Hz, ArH) MS-ESI m/z (% rel lnt ) 275 1 ([MH]⁺, 10), 247 1 (100) 36

Docket 18174 PCT (AP)

HPLC Method A, detection UV 280 nm, SLA 07074 RT = 5 2 mm, peak area 92 %

DL-fhreo-2-Amιno-3-hydroxy-3-(4-methoxyphenyl)-1-(pyrrolιdin-1-yl)propan-1- one hydrochloride SLA 07078

5 To a stirred solution of frans-(4,5-dιhydro-5-(4-methoxyphenyl)oxazol-

4-yl)(pyrrolιdιn-1-yl)methanone SLA 07074 (1 61 g, 5 93 mmol) in methanol (13 mL) was added hydrochloric acid (1mL) After heating at 50 °C for 3 h the mixture reaction was concentrated and the resulting yellow oil was co- evaporated twice with ethyl acetate before solidifying Trituration (ethyl 10 acetate) and drying afforded DL-fhreo-2-amιno-3-hydroxy-3-(4- methoxyphenyl)-1-(pyrrolιdιn-1-yl)propan-1-one hydrochloride SLA 07078 as a white solid (1 64 g, 93 %)

SLA 07078

15 MW 30078, Yield 93 %, white Solid, Mp (⁰C) 1770

¹H-NMR (CD₃OD, δ) 1 32-1 50 (m, 1H, CH₂), 1 50-1 88 (m, 3H, CH₂), 2 15- 2 28 (m, 1H, CH₂N), 3 15-3 42 (m, 4H, 2xCH₂N), 3 79 (s, 3H, CH₃O), 4 06 (d, 1H, J = 9 2 Hz, CH-N), 4 78 (d, 1H, J = 9 2 Hz, CHO), 6 94 (d, 2H, J = 8 5 Hz, ArH), 7 34 (d, 2H, J = 8 5 Hz, ArH)

20 ¹³C-NMR (CD₃OD, δ) 24 8, 26 6, 47 2, 47 6, 559, 59 6, 73 9, 1150 (2xC), 128 9 (2xC), 132 5, 161 7, 1664

DL-tf?reo-2-Amino-1-(4-methoxyphenyl)-3-(pyrrolidtn-1-yl)propan-1-ol Compound 9

To a stirred suspension of DL-fΛreo-[5-(4-methoxy-phenyl)-4,5-dιhydro-

25 oxazol-4-yl]-pyrrolιdιn-1-yl-methanone SLA 07078 (1 61 g, 5 35 mmol) in tetrahydrofuran (200 mL) under nitrogen atmosphere was slowly added, in two portions, lithium aluminium hydride (1 22 g, 32 12 mmol) at 0⁰C The mixture reaction was stirred at RT for 17 h, and then quenched by a slow, T/US2008/054936

Docket 18174 PCT(AP)

dropwise addition of water (50 mL) The white suspension was then concentrated to remove THF and taken back up in a mixture of 300 mL CH₂CI₂ and 1 N aqueous hydrochloric acid (50 mL) The aqueous layer was basified to pH = 10-11 by a stow addition of 1N aqueous sodium hydroxyde 5 The organic layer was removed, combined with additional CH₂CI₂ extracts (4 x 200 mL) and dried over MgSO₄, filtered and evaporated The crude product was purified by column chromatography on silica (CH₂CI₂ MeOH NH₃ = 94 05 01) After evaporation and drying, DL-f/?reo-2-amino-1-(4- methoxyphenyl)-3-(pyrrolιdιn-1-yl)propan-1-ol Compound 9 was obtained (0 10 62 g, 46 %) as a pale yellow solid

OH

Compound 9

MW 250 34, Yield 46 %, Pale Yellow Solid, Mp (⁰C) 77 7

15 Rf 0 35 (CH₂CI₂ MeOH NH₃ = 94 05 01)

¹H-NMR (CDCI₃, δ) 1 65-1 87 (s, 4H, 2xCH₂), 2 40-2 90 (m, 9H, CH₂N, NH₂ & OH), 3 11-3 17 (m, 1H₁ CH₁N), 381 (s, 3H, CH₃O), 4 61 (d, 1H, J = 38 Hz, CH-O), 789 (d 2H, J = 86 Hz, ArH), 726 (d, 2H, J = 8 5 Hz, ArH) 1³C-NMR (CDCI₃, δ) 23 6 (2xC), 54 5, 54 7 (2xC), 55 3, 60 1 , 75 9, 1136,

20 127 4, 1344, 158 8

MS-ESI m/z (% rel Int ) 251 1 ([MH]⁺, 100)

DL-f/)reo-2-(Decanamιdo)-1-(4-methoxyphenyl)-3-(pyrrolιdιn-1-yl)propyl decanoate Compound 10

To a stirred solution of DL-fr>reo-2-amιno-1-(4-methoxy-phenyl)-3-

25 pyrrolιdιn-1-yl-propan-1-ol Compound 9 (0 15 g, 060 mmol) in dichloromethane (10 mL) were added Λ/-hydroxysuccιnιmιde (0 07 g, 0 60 mmol), triethylamine (0 10 mL, 0 63 mmol) and decanoyl chloride (112 μL, 0 54 mmol) under nitrogen atmosphere The mixture reaction was stirred at 008/054936

Docket 18174 PCT (AP)

RT for 22 h and partitioned between methylene chloride and 1 N aqueous sodium hydroxide The organic layer was dried over MgSO₄, filtered and evaporated The crude product was purified by column chromatography on silica (CH₂CI₂ MeOH = 9505) DL-fπreo-2-(Decanamιdo)-1-(4- methoxyphenyl)-3-(pyrrolιdιn-1-yl)propyl decanoate Compound 10 was obtained as a white oil (0 104 g, 31 %)

Compound 10

MW 55884, Yield 40 %, White Oil R_f 0 35 (CH₂CI₂ MeOH = 95 05)

¹H-NMR (CDCI₃, δ) 0 88 (t, 6H, J = 07 Hz, 2xCH₃), 1 26 (s, 14H, 7xCH₂),

1 57-1 59 (m, 4H, 2xCH₂), 1 80 (m, 4H, 2xCH₂), 2 10-2 50 (m, 5H, CH₂), 2 65-

2 76 (m, 5H, CH₂), 3 79 (s, 3H, CH₃O), 454 (m, 1H, CH-N), 5 89 (d, 1H, J = 62 Hz, CH-O), 6 16 (d broad, 1H, J = 8 8 Hz, NH), 6 85 (d, 2H, J = 8 7 Hz, ArH), 7 24 (d, 2H, J = 87 Hz, ArH)

MS-ESI m/z (% rel int ) 559 5 ([MH]⁺, 100)

HPLC Method A, detection UV 280 nm, Compound 10 RT = 6 99 mm, peak area 964 %

Λ/-(DL-fA)reo-1-Hydroxy-1-(4-methoxyphenyl)-3-(pyrrolιdιn-1-yl)propan-2- vDpalmitamide or DL-tf?reo-4-MeO-P4 Compound 11

To a stirred solution of DL-fhreo-2-amιno-1-(4-methoxyphenyl)-3- (pyrrolιdιn-1-yl)propan-1-ol Compound 9 (015 g, 0 60 mmol) in dichloromethane (10 rtiL) were successively added Λ/-hydroxysuccιnιmιde (0 07 g, 060 mmol), tπethylamine (0 100 mL, 0 63 mmol) and palmitoyl chloride (0 15 g, 0 54 mmol) under nitrogen atmosphere The mixture reaction was stirred at RT for 17 h and partitioned between methylene chloride and 1N 8 054936

Docket 18174 PCT (AP)

aqueous sodium hydroxide The organic layer was dried over MgSCU, filtered and evaporated The crude product was purified by column chromatography on silica (CH₂CI₂ MeOH = 95 05) Λ/-(DL-f/ireo-1-Hydroxy-1-(4- methoxyphenyl)-3-(pyrrolιdιn-1-yl)proρan-2-yl)palmιtamιde Compound 11 was obtained as a white solid (0 117 g, 40 %)

Compound 11

MW 488 75, Yield 40 %, White Solid, Mp (⁰C) 82 3 R_f 0 35 (CH₂CI₂ MeOH = 95 05) 10 ¹H-NMR (CDCI₃, 0 88 (t, 3H, J = 7 0 Hz, CH₃), 1 22-1 33 (m, 16H, 8xCH₂),

1 47-1 54 (m, 2H, CH₂), 1 81 (m, 4H, 2xCH₂), 2 09 (t, 2H, J = 7 0 Hz, COCH₂),

2 60-2 80 (m, 4H, 2xCH₂), 2 84 (d, 2H, J = 5 1 Hz, CH₂), 3 80 (s, 3H, CH₃O), 423 (m, 1 H, CH-N), 500 (d, 1 H, J = 22 Hz, CH-O), 5 90 (d, 1 H, J = 74 Hz, NH), 687 (d, 2H, J = 8 7 Hz, ArH), 724 (d, 2H, J = 8 7 Hz, ArH)

15 ¹³C-NMR (CDCI3, δ) 14 1, 22 7, 23 6, 256, 29 1, 29 3, 294, 29 5, 297,

29 7, 31 9, 36 8, 52 3, 55 2, 57 8, 754 113 7 (2xC), 127 0 (2xC), 133 1 ,

158 9, 173 6

MS-ESI m/z (rel int ) 4892 ([MH]⁺, 100)

HP LC Method A, detection UV 280 nm, Compound 11 RT = 6 55 mm, peak 20 area 964 %

DL-f/7reo-2-Amιno-1-(2,3-dιhvdrobenzo[b1[1,41dιoxιn-6-yl)-3-(pyrrolιdιn-1- vDpropan-1 -ol Compound 6

25 frans-(4,5-Dihydro-5-(2,3-dthydrobenzo[6][1 ,4]dioxin-6-yl)oxazol-4- yl)(pyrrolιdin-1-yl)methanone BLE 04100 008/054936

Docket 18174 PCT (AP)

To a stirred and cooled (0⁰C) solution of potassium hydroxide (043 mg, 7 60 mmol) in MeOH (6 5 mL) were added successively 1 ,4-benzodιoxan-6- carboxaldehyde (1 31 g, 796 mmol) and 2-ιsocyano-1-(pyrrolιdιn-1- yl)ethanone BLE 04098 (1 0 g, 657 mmol) The solution was stirred 3 h at 0⁰C and then concentrated The residue was partitioned between EtOAc (100 mL) and water The organic layer was combined with 2 additional EtOAc extracts (2 x 100 mL), washed with brine, dried over MgSO₄, filtered and evaporated Concentration afford to a crude product which was purified by column chromatography on silica (EtOAc) to yield, after evaporation and drying, to fraπs-4,5-dιhydro-5-(2,3-dιhydrobenzo[b][1,4]dιoxιn-6-yl)oxazol-4- yl)(pyrrolιdιn-1-yl)methanone BLE 04100 as a colourless oil (1 76 g, 89 % yield)

BLE 04100

MW 44049 Yield 89 %, colourless oil

¹H-NMR (CDCI₃, δ) 1 75-2 10 (m, 4H, 2xCH₂), 3 40-3 59 (m, 6H, 3xCH₂N), 3 85-4 00 (m, 1H, CHN), 4 26 (s, 4H, CH₂O), 4 59 (dd, 1H, J = 7 5 Hz, J = 2 2 Hz₁ CH-N), 6 00 (d, 1 H, J = 7 5 Hz, CH-O), 6 75-6 90 (m, 3H, ArH), 7 00 (d, 1H, J = 22 Hz, CH=N)

DL-f/7reo-2-amιno-3-(2,3-dιhvdrobenzofb1[1,4ldιoxιπ-6-yl)-3-hvdroxy-1- (pyrrolιdιn-1-yl)propan-1-one hydrochloride Compound 12

To a stirred solution of frar?s-4,5-dιhydro-5-(2,3- dιhydrobenzo[ib][1 ,4]dιoxιn-6-yl)oxazol-4-yl)(pyrrolιdιn-1 -yl)methanone BLE 04100 (1 74 g, 5 77 mmol) in methanol (15 mL) was added hydrochloric acid (1mL) After heating at 50°C for 3h the mixture reaction was concentrated and the resulting yellow oil was co-evaporated twice with ethyl acetate before 2008/054936

Docket 18174 PCT (AP)

solidifying Trituration (ethyl acetate) and drying afforded DL-f/ireo-2-amino-3- (2,3-dιhydrobenzo[to][1,4]dιoxιn-6-yl)-3-hydroxy-1-(pyrrolιdιtv1-yl)propan-1- one hydrochloride Compound 12 as a white solid (1 85 g, 95 %)

5

Compound 12

MW 328 79, Yield 95 0 %, White Solid, Mp (⁰C) 1762 1H-NMR (CD₃OD, δ) 1 42-1 58 (m, 1H, CH₂), 1 58-1 70 (m, 1H, CH₂), 1 70- 1 88 (m, 2H, CH₂), 320-345(m, 4H, N-CH₂), 406 (d, 1 H, J = 9 1 Hz, CH-N), 10 425 (s, 2H, CH₂), 4 75 (d, 1 H, J = 92 Hz, CH-O), 4 89 (s, 2H, CH₂), 6 82-6 95 (m, 3H, ArH)

¹³C-NMR (CD₃OD, δ) 24 9, 26 7, 47 3, 47 6, 59 5, 657, 73 6, 1164, 118 3, 120 3, 1337, 145 1, 145 6, 1664

15 DL-f/?reo-2-Amιno-1-(2,3-dιhydrobenzorbH1.4Tdιoxιn-6-yl)-3-(pyrrolιdιn-1- yl)propan-1-ol Compound 6

To a stirred suspension of fraπs-(4,5-dιhydro-5-(4- methoxyphenyl)oxazol-4-yl)(pyrrolιdιn-1-yl)methanone SLA 07080 (1 79 g, 544 mmol) in THF (220 mL) was slowly added at 0 °C, in two portions, L1AIH4

20 (1 28 g, 337 mmol) The mixture was stirred at RT for 3 5 h and quenched by a slow addition of water at 0 °C (350 mL) The white suspension was concentrated to remove THF and taken back in a mixture of CH₂CI₂ (300 mL) and 1 N aqueous HCI (50 mL) The aqueous layer was basified to pH = 10-11 by slow addition of 1 N aqueous NaOH The organic layer was removed, two

25 more extracts were combined and dried over MgSO4, filtered and evaporated Concentration afforded to a crude product as a yellow oil This material was purified by column chromatography on silica (CH₂CI₂ MeOH NH4OH 20% =

94 5 1) to led to DL-tf?reo-2-amιno-1-(2,3-dιhydrobenzo[6][1 4]dιoxιn-6-yl)-3- T US2008/054936

Docket 18174 PCT (AP)

(pyrrolιdιn-1-yl)propan-1-ol Compound 6 (0 705 g, 46 5 % yield) as a near colorless gum

OH

^._QA^ NH₂ ^ — /

(+/-)

5 Compound 6

MW 278 35, Yield 46 5 %, Colorless Gum R_f 0 20 (CH₂CI₂ MeOH NH₄OH 20 % = 94 5 1)

¹H-NMR (CDCI₃, δ) 1 70-1 85 (m, 4H, 2xCH₂), 2 40-2 70 (m, 6H, 3xCH₂N-), 3 05-3 15 (m, 1H, CH-N), 4 25 (s, 4H, CH₂O), 4 55 (d, 1H, J = 2 2 Hz, CH-O), 10 5 30 (s, 1H, -OH), 675-690 (m, 3H, ArH)

A/-(DL-fhreo-1-(2,3-dιhvdrobenzo[b1H ,41dιoxιn-6-ylH-hvdroxy-3-(pyrrolιdιn-1- yl)propan-2-yl)decanamιde Compound 7

To a stirred solution of DL-tfjreo-2-amιno-1-(2,3-

15 dthydrobenzo[b][1,4]dιoxιn-6-yl)-3-(pyrrolιdιn-1-yl)propan-1-ol BLE 04104 (0 186 g, 0 67 mmol) in 10 mL CH2CI2 were added, in order, N- hydroxysuccinimide (0 081 g, 0 70 mmol) in 2 mL CH2CI2, triethylamine (112 μL, 0 80 mmol) and decanoyl chloride (125 μL, 060 mmol) The mixture was stirred overnight at RT and then partitioned between CH₂CI₂ and 1 N aqueous

20 sodium hydroxide The organic layer was dried over MgSO₄, filtered and evaporated and the residue obtained was purified by column chromatography on silica (CH₂CI₂ MeOH = 95 5) A white solid Λ/-(DL-frireo-1-(2,3- dιhydrobenzo[b][1,4]dιoxιn-6-yl)-1-hydroxy-3-(pyrrolιdιn-1-yl)propan-2- yl)palmιtamιde Compound 7 was obtained (126 mg, 43 5 % yield)

25 Docket 18174 PCT (AP)

Compound 7

MW 516 76, Yield 43 5 %, White Solid, Mp (⁰C) 84 6 R_f 040 (MeOH CH₂CI₂ = 10 90) 5 ¹H-NMR (CDCI₃, δ) 0 88 (t, 3H, J = 6 7 Hz, CH₃), 1 12-1 39 (m 12 H), 1 40-

1 60 (m, 2H, CH2), 1 72-1 90 (m, 4H, 2xCH2), 2 10 (t, 2H, J = 6 7 Hz, CH2),

2 55-2 90 (m, 6H), 4 13-4 30 (m, 1H, CH-N), 4 24 (s, 4H, CH2N), 4 91 (d, 1H, J = 3 3 Hz, CH-O), 5 90 (d, 1H, J = 7 4 Hz, NH), 6 75-6 88 (m, 3H, ArH), OH not seen

10 ¹³C-NMR (CDCI₃, δ) 14 1, 22 7, 236 (2xC), 25 6, 29 1 , 29 3, 31 9, 36 8,

52 3, 55 1 (2xC), 57 7, 64 3 (2xC), 75 2, 77 2, 115 0, 117 0, 118 9, 134 4,

142 8, 1434, 173 5, 174 8

MS-ESI m/z (% rel Int ) 433 1 ([MH]⁺, 100)

HPLC Method A, detection UV 280 nm, Compound 7, RT = 5 2 mm, peak 15 area 96 2 %

Λ/-(DL-f/7reo-1-(2,3-D[hvdrobenzo[biπ,41dιoxιn-6-ylV1-hvdroxy-3-(pyrrolιdιn-1- yl)propan-2-yl)palmιtamιde Compound 8

To a stirred solution of DL-fhreo-2-amιno-1-(2,3-

20 dιhydrobenzo[ό][1,4]dιoxιn-6-yl)-3-(pyrrolιdιn-1-yl)propan-1-ol BLE 04104 (0 158 g, 0 57 mmol) in 10 mL CH₂CI₂ were added, in order, N- hydroxysucαnimide (0068 g, 0 59 mmol) in 2 ml CH2CI2, triethylamine (95 μL, 068 mmol) and palmitoyl chloride (155 μL, 0 511 mmol) in 3 mL CH₂CI₂ The mixture was stirred overnight at RT and then partitioned between CH₂CI₂

25 and 1 N aqueous sodium hydroxyde The organic layer was purified by column chromatography on silica using as eluent CH₂CI₂ MeOH = 95 5 A white solid N-(DL-tf)reo-1-(2,3-dιhydrobenzo[b][1 ,4]dιoxιn-6-yl)-1-hydroxy-3- US2008/054936

Docket 18174 PCT (AP)

(pyrrolιdιn-1-yl)propan-2-yl)palmιtamιde Compound 8 was obtained (148 mg, 504 % yield)

Compound 8

5

MW 516 7, Yield 504 %, White Solid, Mp (°C) 664

R_f 0 50 (MeOH CH₂CI₂ = 10 90)

¹H-NMR (CDCI₃, δ) 0 88 (t, 3H, J = 6 7 Hz, CH₃), 1 15-1 35 (m, 24 H), 1 45-

158 (m, 2H, CH₂), 175-190 (rπ, 4H, 2xCH₂), 210 (t, 2H₁J= 7 A Hz, CH₂), 10 261 (s, 1H, OH), 252-272 (m, 4H), 272-292 (m, 2H), 415-422 (m, 1H, CH-

N), 424 (s, 4H, CH₂N), 492 (d, 1H, J= 33 Hz, CH-O), 608 (d, 1H, J= 74

Hz, NH), 675-690 (m, 3H, ArH)

MS-ESI m/z (% rel lnt ) 517 2 ([MH]⁺, 100)

HPLC Method A, detection UV 280 nm, Compound 8 RT = 660 mm, peak 15 area 972 %

Preparation of DL-f/7reo-2-Amιno-1-(pyrιdm-4-yl)-3-(pyrrolιdm-1-yl)propan-1-ol Compound 46 Trans-(4,5-dιhydro-5-(pyrιdιn-3-yl)oxazol-4-yl)(pyrrolιdιn-1-yl)methanone BLE

20 04110B

A general method D for oxazohnes formation is illustrated by the preparation of BLE 04110B To a stirred and cooled (0°C) solution of potassium hydroxide (0 55 g, 9 80 mmol) in methanol (10 mL) were added a mixture of 3-pyrιdιne carboxaldehyde (1 03 mL, 10 84 mmol) and 2-ιsocyano-

25 1-(pyrrolιdιn-1-yl)ethanone BLE 04098 (1 50 g, 10 86 mmol) The solution was stirred 3 h at 0°C and then concentrated The residue was partitioned between ethyl acetate (100 mL) and water The organic layer was combined T/US2008/054936

Docket 18174 PCT (AP)

with two additional ethyl acetate extracts (2x100 mL), washed with aqueous sodium chloride and dried over MgSO₄, filtered and evaporated Concentration afforded a crude product which was purified by column chromatography on silica (CH₂CI₂ MeOH = 98 2) to yield to frans-(4,5-dιhydro- 5-(pyπdιn-3-yl)oxazol-4-yl)(pyrrolιdιn-1-yl)methanone BLE 0411OB (0 95 g, 39 %) as a pale yellow pale solid

BLE 0411OB MW 245 28, Yield 39 % Yellow Pale Solid, Mp (⁰C) 107 0 1H-NMR (CDCI₃, δ) 1 78-2 10 (m, 4H, 2xCH₂), 340-361 (m, 3H CH₂N), 3 90-4 04 (m, 1 H, CH₂N), 4 59 (dd, 1 H, J = 7 7 Hz, J = 2 2 Hz, CH-N), 621 (d, 1 H, J = 7 7 Hz, CH-O), 7 04 (d, 1H, J = 2 2 Hz, 0-CH=N), 7 33 (m, 1H, ArH), 7 64 (m, 1H, ArH), 8 59 (d, 2H, J = 2 8 Hz, ArH) 1³C-NMR (CDCI₃, δ) 242, 260, 464, 466, 757, 79 3, 123 7, 133 5, 135 3, 147 6, 149 9, 1552, 1662 frans-(4,5-Dιhvdro-5-(pyrιdιn-4-yl)oxazol-4-yl)(pyrrolιdιn-1-yl)methanone Compound 19

Compound 19 was prepared in accordance with method D using pyrιdιne-4-carbaldehyde (1 88 mL, 1976 mmol), KOH (1 01 g, 18 00 mmol) in methanol (18 mL) and 2-ιsocyano-1-(pyrrolιdιn-1-yl)ethanone BLE 04098

(2 73 g, 19 76 mmol) The residue was partitioned between ethyl acetate (200 mL) and water (150 mL) The organic layer was combined with additional ethyl acetate extracts (2 x 150 mL), washed with aqueous sodium chloride (2 x 150 mL) and dried over MgSO₄, filtered and evaporated Trans-(4,5-dιhydro- 5-(pyrιdιn-4-yl)oxazol-4-yl)(pyrrolιdιn-1-yl)methanone Compound 19 was obtained as a white solid (4 32 g, 98 % yield) Docket 18174 PCT (AP)

Compound 19

MW 245 28, Yield 98 %, White Solid, Mp (⁰C) = 69 2 R_f 0 65 (MeOH CH₂CI₂ = 1090) 1H-NMR (CDCI₃, δ) 1 78-206 (m, 4H, 2xCH₂), 344-360 (m, 3H, CH₂N),

3 90-4 01 (m, 1H, CH₂N), 4 52 (dd, 1H, J - 1 9 Hz, J = 2 2 Hz, CH-N), 6 19 (d, J = 7 9 Hz, 1H, CH-O), 703 (d, 1H, J = 22 Hz, N=CH-O), 724 (dd, 2H, J =

45 Hz, J = 1 5 Hz, ArH), 8 61 (dd, 2H, J = 4 5 Hz, J = 1 5 Hz, ArH)

A general method for the acidic hydrolysis of oxazolines (Method E) is illustrated in the preparation of Compound 20 which is a substituted propionic acid amide and is made from the oxazolme intermediate BLE 0411OB which can be prepared in accordance with General Synthetic Scheme 1

BLE 04110B

2008/054936

Docket 18174 PCT (AP)

DL-tf7rβo-2-Amιno-3-hvdroxy-3-(pyrιdιn-3-yl)-1-(pyrrolιdιn-1-yl)propan-1-one dihydrochlonde Compound 20

To a solution of frans-(4,5-dιhydro-5-(pyπdιn-3-yl)oxazol-4- yl)(pyrrolιdιn-1-yl)methanone BLE 0411OB (0 932 g, 3 80 mmol) in methanol

5 (10 mL) was added hydrochloric acid 37 % (1 2 mL) After heating (50 ⁰C) the mixture for 225 h the reaction mixture was concentrated and the crude product was coevaporated twice with ethyl acetate After trituration with ethyl acetate, filtration and drying DL-fl?reo-2-amιno-3-hydroxy-3-(pyrιdιn-3-yl)-1- (pyrrohdιn-1-yl)propan-1-one dihydrochlonde Compound 20 was obtained as

10 a white solid (1 10 g, 94 % yield)

Compound 20

MW 308 2, Yield 94 %, White Solid, Mp (⁰C) 1234

¹H-NMR (CD₃OD, δ) 1 65-200 (m, 4H, 2xCH₂), 2 82-3 11 (m 1 H, -CH₂N), 15 3 30-357 (m, 2H, CH₂N), 3 57-3 77 (m, 1H, CH₂N), 4 54 (d, 1H, J = 5 3 Hz,

CH-N), 5 38 (d, 1H, J = 5 3 Hz, CH-O), 8 15 (dd, 1H, J = 7 6 Hz, J = 5 0 Hz,

ArH), 8 68 (d, 1H, J = 7 6 Hz, ArH), 8 89 (d, 1 H, J = 7 6 Hz, ArH), 8 96 (s, 1H,

ArH)

¹³C-NMR (CD₃OD₁) 24 9, 269, 477, 482, 58 1 , 696, 1287, 141 5, 141 6, 20 143 1, 146 5, 1654

DL-f/7reo-2-Ammo-3-hvdroxy-3-(pyrιdιπ-4-yl)-1-(pyrrolιdιn-1-v0propan-1-one dihvdrochloride Compound 22

Compound 22 was prepared following method E with trans-(A 5- dιhydro-5-(pyrιdιn-4-yl)oxazol-4-yl)(pyrrolιdιn-1-yl)methanone Compound 19 25 (0 750 g, 307 mmol), hydrochloric acid 37 % (1 0 mL) and methanol (10 mL)

After 30 h at 50 ⁰C and work-up DL-#?reo-2-amιno-3-hydroxy-3-(pyπdιn-4-yl)-

1-(pyrrolιdιn-1-yl)propan-1-one dihydrochlonde Compound 22 was obtained as a white solid (0 935 g, 99 %) 2008/054936

Docket 18174 PCT (AP)

Compound 22

MW 30828, Yield 99 %, White Solid, Mp (X) 1170

¹H-NMR (CD₃OD, δ) 1 75-2 03 (m, 4H, 2xCH₂), 2 93-3 08 (m, 1H₁ CHN), 3 32-3 75 (m, 3H, 2xCH₂), 4 54 (d, 1H, J = 5 9 Hz, CH₁N), 540 (d, 1H, J = 5 9 Hz, CH-O), 8 21 (d, 2H, J = 5 8 Hz, ArH), 8 94 (d, 2H, J = 5 8 Hz, ArH) MS-ESI m/z (% rel int ) 236 1 ([MH]⁺, 17), 219 (25), 148 (100) HPLC Method A, detection UV 254 nm, Compound 22 RT = 0 8 mm, peak area 963 % DL-tfyeo-2-Amιno-1-(pyπdιn-4-viy3-(pyrrolιdιn-1-yl)propan-1-ol Compound 46 To a stirred suspension of DL-tfjreo-2-amιno-3-hydroxy-3-(pyrιdιn-4-yl)- 1-(pyrrolιdιn-1-yl)propan-1-one dihydrochloπde Compound 22 (0 86 g, 2 80 mmol) in tetrahydrofuran (108 mL) under nitrogen atmosphere was slowly added, in two portions, lithium aluminium hydride (064 g, 1682 mmol) at 0 °C The mixture reaction was stirred at RT for 20 h and quenched by a slow, dropwise addition of 2 N aqueous sodium hydroxyde (84 mL, 6 eq) The yellow precipitate was filtered The organic layer was washed by water (80 mL) and the organic layer was removed and combined with additional ethyl acetate extracts (4 x 200 mL) and dried over MgSO₄, filtered and evaporated The crude product was purified by column chromatography on silica

(CH₂CI₂ MeOH NH₃ = 94 05 01) After evaporation and drying DL-threo-2- amιno-1-(pyrιdιn-4-yl)-3-(pyrrolιdιn-1-yl)propan-1-ol Compound 46 was obtained (0 075 g, 12 %) as a pale yellow solid

OH

NH₂ ^- -

Compound 46 Docket 18174 PCT (AP)

MW 221 30, Yield 12 %, Pale Yellow Solid

R_f 0 35 (CH₂CI₂ MeOH NH₃ = 90 0802)

¹H-NMR (CD₃OD, δ) 1 60-1 80 (m, 4H, 2xCH₂), 2 30-2 80 (m, 6H, 3xCH₂N),

3 14-3 19 (m, 1H, CH₁NH₂), 468 (d, 1H, J = 3 0 Hz, CH-O), 730 (d, 2H, J =

60 Hz, ArH), 8 55 (d, 2H, J = 60 Hz, ArH)

¹³C-NMR (CD₃OD, δ) 23 5 (2xC), 54 1, 547 (2xC), 60 1, 74 5, 121 4 (2xC),

1495 (2xC), 152 1

MS-ESI m/z (rel int ) 222 1 ([MH]⁺, 100), 205 0 (80), 189 0 (45), 151 0 (70),

134 0 (42), 121 9 (100), 107 9 (40)

Claims

36Docket 18174 PCT (AP)WHAT IS CLAIMED IS:

1. The use of a compound in the manufacture of a medicament for treating a cognitive disorder, wherein the compound has the following structure:

Formula 2 wherein Ri is H or alkyl of 1 to 6 carbons,

R₂ is H, alkyl of 1 to 6 carbons or the Ri and R₂ groups together with the nitrogen form a saturated or unsaturated 4, 5, 6 or 7 membered ring that optionally includes one or two heteroatoms independently selected from N, O and S₁ said 4, 5, 6 or 7 membered ring optionally being substituted with a halogen or with an alkyl group having 1 to 6 alkyl groups; R₃ is independently selected from H, alkyl of 1 to 20 carbons, aryl or heteroaryl, aryl-alkyl or heteroaryl-alkyl wherein the alkyl moiety is has 1 to 4 carbons, cycloalkyl of 3 to 6 carbons, said aryl or heteroaryl groups being optionally substituted with 1 to 3 groups independently selected from the group consisting of halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons, or

R₃ is CO-R₇ or CO-O-R₇ wherein R₇ is H, alkyl of 1 to 1 to 20 carbons, benzyl, alkyl of 1 to 20 carbons substituted with and NH₂ group, with a NHCOOalkyl or with an NH-COalkyl group wherein the alkyl group has one to 6 carbons, or R₇ is aryl, heteroaryl, aryl-alkyl or heteroaryl-alkyl wherein the alkyl moiety is branched or unbranched and has 1 to 4 carbons, said aryl or heteroaryl groups being optionally substituted with 1 to 3 groups independently selected from the group consisting of halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons;

R₄ is H, alkyl of 1 to 6 carbons or CO-Rs wherein Rs is alkyl of 1 to 6 carbons; Docket 18174 PCT (AP)

the wavy lines represent bonds connected to carbons having R or S configuration, and

Rio is selected from the groups of formulas (i) and (ii)

0) (H) wherein the ^* indicates the carbon atom to which the remaining moiety of the molecule is attached;

R₅ and Re independently are H, alkyl of 1 to 6 carbons, halogen, alkoxy of 1 to 6 carbons or the R₅ and R₆ groups together with the atoms to which they are attached jointly form a carbocyclic or a heterocyclic ring, the carbocyclic ring having 5 or 6 atoms in the ring, the heterocyclic ring having 5 or 6 atoms in the ring and 1 to 3 heteroatoms independently selected from N, O and S, and said carbocyclic or heterocyclic ring jointly formed by R₅ and Re being optionally substituted with 1 to 6 Rg groups wherein R₉ is independently selected from halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons, or a pharmaceutically acceptable salt of said compound; with the proviso: that when Rio has formula (ii) then the claim does not include compounds wherein R₄ is hydrogen and Ri and R₂ jointly with the nitrogen form a morpholin or a pyrrolidin ring and wherein R₅ and R₆ both are H or one of R₅ and Re is OCH₃ and the other is H.

2. The use according to claim 1 , wherein R₁₀ represents the formula (i).

3. The use according to claim 1 , wherein R₁₀ represents the formula (ii). Docket 18174 PCT (AP)

4. The use according to claim 2, wherein the compound has the formula

DL-threo wherein R₅ and R_& are independently selected from H, alkyl and alkoxy and R₄ is H or CO-R₈ or a pharmaceutically acceptable salt of said compound.

5. The use according to Claim 4, wherein the compound has the formula

OH

N^ NH₂ ^V

DL-threo or a pharmaceutically acceptable salt of said compound.

6. The use according to claim 3, wherein the compound has the formula

DL-threo wherein R₃ is CO-R₇ or CO-O-R₇, R₄ is CO-R₈ and R₅ and R₆ are independently selected from H, alkyl of 1 to 6 carbons and alkoxy of 1 to 6 carbons or any other pharmaceutically acceptable salt of said compound.

7. The use according to Claim 6, wherein the compound has the formula Docket 18174 PCT (AP)

HCI l-threo or any other pharmaceutically acceptable salt of said compound.

8. The use of a compound in the manufacture of a medicament for treating a cognitive disorder, wherein the compound has the following structure:

or any other pharmaceutically acceptable salt of said compound.

9. The use of a compound in the manufacture of a medicament for treating a cognitive disorder, wherein the compound has the following structure

L-threo or any other pharmaceutically acceptable salt of said compound.

10. The use according to any of the preceding claims, wherein the cognitive disorder is selected from the group consisting of an agnosia, an amnesia, an aphasia, an apraxia, a delirium, a dementia, and a learning disorder. Docket 18174 PCT (AP)

11. The use according to claim 10, wherein the cognitive disorder is selected from the group consisting of AIDS dementia complex, Binswanger's disease, dementia with Lewy Bodies, frontotemporal dementia, mild cognitive impairment, multi-infarct dementia, Pick's disease, semantic dementia, senile dementia, and vascular dementia.

12. The use according to claim 10, wherein the learning disorder is selected from the group consisting of Asperger's syndrome, attention deficit disorder, attention deficit hyperactivity disorder, autism, childhood disintegrative disorder, and Rett syndrome.

13. The use according to claim 10, wherein the aphasia is progressive non- fluent aphasia.

14. The use according to any of the preceding claims, wherein the cognitive disorder is associated with neurodegenerative disease, injury to the brain, psychiatric disorders, or chronic pain.

15. The use according to claim 14, wherein the neurodegenerative disease is selected from the group consisting of Alzheimer's disease, corticobasal degeneration, Creutzfeldt-Jacob disease, frontotemporal lobar degeneration, Huntington disease, multiple sclerosis, normal pressure hydrocephalus, organic chronic brain syndrome, Parkinson's disease, Pick disease, progressive supranuclear palsy, and senile dementia (Alzheimer type).

16. The use according to claim 14, wherein the injury to the brain is selected from the group consisting of chronic subdural hematoma, concussion, intracerebral hemorrhage, encephalitis, meningitis, septicemia, drug intoxication, and drug abuse. Docket 18174 PCT (AP)

17. The use according to claim 14, wherein the psychiatric disorders are selected from the group consisting of anxiety disorders, dissociative disorders, mood disorders, schizophrenia, and somatoform and factitious disorders.