Patent Grant
7799785
This application is the national stage of Application No. PCT/EP2005/056544, filed 6 Dec. 2005, which application claims priority from PCT/EP04106373.6 filed 7 Dec. 2004.
This invention concerns novel substituted tetracyclic tetrahydrofuran, pyrrolidine and tetrahydrothiophene derivatives with binding affinities towards serotonin receptors, in particular 5-HT2A and 5-HT2C receptors, and towards dopamine receptors, in particular dopamine D2 receptors and with norepinephrine reuptake inhibition properties, pharmaceutical compositions comprising the compounds according to the invention, the use thereof as a medicine, in particular for the prevention and/or treatment of a range of psychiatric and neurological disorders, in particular certain psychotic, cardiovascular and gastrokinetic disorders and processes for their production.
WO 97/38991, published Oct. 23, 1997 (Janssen Pharmaceutica N.V.) discloses substituted tetracyclic tetrahydrofuran derivatives that may be used as therapeutic agents in the treatment or prevention of CNS disorders, cardiovascular disorders or gastrointestinal disorders. In particular, the compounds show affinity for the serotonin 5-HT2 receptors, particularly for the 5-HT2A and 5-HT2C-receptors.
WO 99/19317, published Apr. 22, 1999 (Janssen Pharmaceutica N.V.) discloses substituted tetracyclic tetrahydrofuran derivatives with a specific halogen substitution pattern on the dibenzoazepine, dibenzooxepine, dibenzothiepine or dibenzosuberane ring. The compounds are useful in the treatment or prevention of CNS disorders, cardiovascular disorders or gastrointestinal disorders and show a faster onset of action over the compounds as disclosed in WO 97/38991.
Both WO 03/048146, published Jun. 12, 2003 (Janssen Pharmaceutica N.V.) and WO 03/048147, published Jun. 12, 2003 (Janssen Pharmaceutica N.V.) disclose processes for the preparation of each of the 4 diastereomers of cis-, respectively transfused 3,3a,8,12b-tetrahydro-2H-dibenzo[3,4:6,7]cyclohepta[1,2-b]furan derivatives in a stereochemically pure form from a single enantiomerically pure precursor. The compounds show affinity for the serotonin 5-HT2A, 5-HT2C and 5-HT7 receptors and the H1-receptors (pIC50=7.15−7.89), D2 and/or D3 receptors and for the norepinephrine reuptake transporters (PIC50=6.01−7.34).
WO 03/040122, published May 15, 2003 (Janssen Pharmaceutica N.V.) discloses mandelate salts of the compounds according to WO 97/38991 and WO 99/19317. Said salts were surprisingly found to be more stable at enhanced temperature and relative humidity than the compounds disclosed in WO 97/38991 and WO 99/19317.
It is the object of the present invention to provide novel analogues of the tetracyclic tetrahydrofuran derivatives of WO 97/38991 and WO 99/19317, which differ from such derivatives in that they demonstrate in general more selectivity for the norepinephrine reuptake transporter than the 5-HT2A, 5-HT2C and dopamine D2 receptors, resulting in compounds which have a more pronounced antidepressant effect in relation to their antipsychotic properties. The compounds of formula (I) below where the basic nitrogen atom at the C-2 position is embedded in a cyclic system demonstrate a potent antagonistic effect against the 5-HT2A, 5-HT2C and dopamine D2 receptors.
This goal is achieved by the present novel compounds according to Formula (I):
an N-oxide form, a pharmaceutically acceptable addition salt or a stereochemically isomeric form thereof, wherein:
More in particular, the invention relates to a compound according to Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form thereof and a prodrug thereof, wherein A and B are each benzo, optionally substituted with fluoro. Preferably, A is unsubstituted and B is substituted with fluoro at the 11-position.
More in particular, the invention relates to a compound according to Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form thereof and a prodrug thereof, wherein C is a group of formula (c-1) or (c-2); wherein
More in particular, the invention relates to a compound according to Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form thereof and a prodrug thereof, wherein C is a group of formula (c-3) or (c-4); wherein
More in particular, the invention relates to a compound according to Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form thereof and a prodrug thereof, wherein (d-1) is defined as wherein:
More in particular, the invention relates to a compound according to the general Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form thereof and a prodrug thereof, wherein:
Preferably, alkyl is methyl, ethyl or propyl, optionally substituted with one or more halo, cyano, oxo, hydroxy, formyl, carboxyl or amino radicals. Preferably, alkyl is optionally substituted with hydroxy.
Preferably, aryl is phenyl, optionally substituted with 1, 2 or 3 substituents selected from the group of halo, nitro, cyano, hydroxy, alkyloxy or alkyl. Preferably, aryl is unsubstituted.
Preferably, halo is fluoro.
Preferred compounds are also those particular compounds according to the invention wherein the hydrogen atoms on carbon atoms 3a and 12b have a trans configuration and those having the (2α, 3aα, 12bβ) stereochemical configuration.
Most preferred compounds are also those compounds according to the invention where the compounds are selected from the group of compounds defined by the compound numbers given in Tables 1 to 4.
In the framework of this application, alkyl is defined as a monovalent straight or branched saturated hydrocarbon radical having from 1 to 6 carbon atoms, for example methyl, ethyl, propyl, butyl, 1-methylpropyl, 1,1-dimethylethyl, pentyl and hexyl; alkyl further defines a monovalent cyclic saturated hydrocarbon radical having from 3 to 6 carbon atoms, for example cyclopropyl, methylcyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. The definition of alkyl also comprises an alkyl radical that is optionally substituted on one or more carbon atoms with one or more phenyl, halo, cyano, oxo, hydroxy, formyl and amino radicals, for example hydroxyalkyl, in particular hydroxymethyl and hydroxyethyl and polyhaloalkyl, in particular difluoromethyl and trifluoromethyl.
In the framework of this application, halo is generic to fluoro, chloro, bromo and iodo.
In the framework of this application, with “compounds according to the invention” is meant a compound according to the general Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form thereof and a prodrug thereof.
In the framework of this application, an element, in particular when mentioned in relation to a compound according to Formula (I), comprises all isotopes and isotopic mixtures of this element, either naturally occurring or synthetically produced, either with natural abundance or in an isotopically enriched form. In particular, when hydrogen is mentioned, it is understood to refer to 1H, 2H, 3H and mixtures thereof; when carbon is mentioned, it is understood to refer to 11C, 12C, 13C, 14C and mixtures thereof; when nitrogen is mentioned, it is understood to refer to 13N, 14N, 15N and mixtures thereof; when oxygen is mentioned, it is understood to refer to 14O, 15O, 16O, 17O, 18O and mixtures thereof; and when fluor is mentioned, it is understood to refer to 18F, 19F and mixtures thereof.
The compounds according to the invention therefore also comprise compounds with one or more isotopes of one or more element, and mixtures thereof, including radioactive compounds, also called radiolabelled compound, wherein one or more non-radioactive atoms has been replaced by one of its radioactive isotopes. By the term “radiolabelled compound” is meant any compound according to Formula (I), an N-oxide form, a pharmaceutically acceptable addition salt or a stereochemically isomeric form thereof, which contains at least one radioactive atom. For example, compounds can be labelled with positron or with gamma emitting radioactive isotopes. For radioligand-binding techniques (membrane receptor assay), the 3H-atom or the 125I-atom is the atom of choice to be replaced. For imaging, the most commonly used positron emitting (PET) radioactive isotopes are 11C, 18F, 15O and 13N, all of which are accelerator produced and have half-lives of 20, 100, 2 and 10 minutes respectively. Since the half-lives of these radioactive isotopes are so short, it is only feasible to use them at institutions which have an accelerator on site for their production, thus limiting their use. The most widely used of these are 18F, 99mTc, 201Tl and 123I. The handling of these radioactive isotopes, their production, isolation and incorporation in a molecule are known to the skilled person.
In particular, the radioactive atom is selected from the group of hydrogen, carbon, nitrogen, sulfur, oxygen and halogen. Preferably, the radioactive atom is selected from the group of hydrogen, carbon and halogen.
In particular, the radioactive isotope is selected from the group of 3H, 11C, 18F, 122I, 123I, 125I, 131I, 75Br, 76Br, 77Br and 82Br. Preferably, the radioactive isotope is selected from the group of 3H, 11C and 18F.
The pharmaceutically acceptable salts are defined to comprise the therapeutically active non-toxic acid addition salt forms that the compounds according to Formula (I) are able to form. Said salts can be obtained by treating the base form of the compounds according to Formula (I) with appropriate acids, for example inorganic acids, for example hydrohalic acid, in particular hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and phosphoric acid; organic acids, for example acetic acid, hydroxyacetic acid, propanoic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid, mandelic acid, fumaric acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclamic acid, salicylic acid, p-aminosalicylic acid and pamoic acid.
The compounds according to Formula (I) containing acidic protons may also be converted into their therapeutically active non-toxic metal or amine addition salts forms by treatment with appropriate organic and inorganic bases. Appropriate base salts forms comprise, for example, the ammonium salts, the alkaline and earth alkaline metal salts, in particular lithium, sodium, potassium, magnesium and calcium salts, salts with organic bases, e.g. the benzathine, N-methyl-D-glucamine, hybramine salts, and salts with amino acids, for example arginine and lysine.
Conversely, said salts forms can be converted into the free forms by treatment with an appropriate base or acid.
The term addition salt as used in the framework of this application also comprises the solvates that the compounds according to Formula (I) as well as the salts thereof, are able to form. Such solvates are, for example, hydrates and alcoholates.
The N-oxide forms of the compounds according to Formula (I) are meant to comprise those compounds of Formula (I) wherein one or several nitrogen atoms are oxidized to the so-called N-oxide, particularly those N-oxides wherein one or more tertiary nitrogens (e.g of the piperazinyl or piperidinyl radical) are N-oxidized. Such N-oxides can easily be obtained by a skilled person without any inventive skills and they are obvious alternatives for the compounds according to Formula (I) since these compounds are metabolites, which are formed by oxidation in the human body upon uptake. As is generally known, oxidation is normally the first step involved in drug metabolism (Textbook of Organic Medicinal and Pharmaceutical Chemistry, 1977, pages 70-75). As is also generally known, the metabolite form of a compound can also be administered to a human instead of the compound per se, with much the same effects.
The compounds according to the invention possess at least 1 oxydizable nitrogen (tertiary amines moiety). It is therefore highly likely that N-oxides are to form in the human metabolism.
The compounds of Formula (I) may be converted to the corresponding N-oxide forms following art-known procedures for converting a trivalent nitrogen into its N-oxide form. Said N-oxidation reaction may generally be carried out by reacting the starting material of Formula (I) with an appropriate organic or inorganic peroxide. Appropriate inorganic peroxides comprise, for example, hydrogen peroxide, alkali metal or earth alkaline metal peroxides, e.g. sodium peroxide, potassium peroxide; appropriate organic peroxides may comprise peroxy acids such as, for example, benzenecarboperoxoic acid or halo substituted benzenecarboperoxoic acid, e.g. 3-chlorobenzenecarboperoxoic acid, peroxoalkanoic acids, e.g. peroxoacetic acid, alkylhydroperoxides, e.g. tert-butyl hydroperoxide. Suitable solvents are, for example, water, lower alkanols, e.g. ethanol and the like, hydrocarbons, e.g. toluene, ketones, e.g. 2-butanone, halogenated hydrocarbons, e.g. dichloromethane, and mixtures of such solvents.
The term “stereochemically isomeric forms” as used hereinbefore defines all the possible isomeric forms that the compounds of Formula (I) may possess. Unless otherwise mentioned or indicated, the chemical designation of compounds denotes the mixture of all possible stereochemically isomeric forms, said mixtures containing all diastereomers and enantiomers of the basic molecular structure. More in particular, stereogenic centers may have the R- or S-configuration; substituents on bivalent cyclic (partially) saturated radicals may have either the cis- or trans-configuration. Compounds encompassing double bonds can have an E or Z-stereochemistry at said double bond. Stereochemically isomeric forms of the compounds of Formula (I) are obviously intended to be embraced within the scope of this invention.
Following CAS nomenclature conventions, when two stereogenic centers of known absolute configuration are present in a molecule, an R or S descriptor is assigned (based on Cahn-Ingold-Prelog sequence rule) to the lowest-numbered chiral center, the reference center. R* and S* each indicate optically pure stereogenic centers with undetermined absolute configuration. If “α” and “β” are used: the position of the highest priority substituent on the asymmetric carbon atom in the ring system having the lowest ring number, is arbitrarily always in the “α” position of the mean plane determined by the ring system. The position of the highest priority substituent on the other asymmetric carbon atom in the ring system (hydrogen atom in compounds according to Formula (I)) relative to the position of the highest priority substituent on the reference atom is denominated “α”, if it is on the same side of the mean plane determined by the ring system, or “β”, if it is on the other side of the mean plane determined by the ring system.
The numbering of the tetracyclic ring-systems present in the compounds of Formula (I-a) and (I-b) when A and B are benzo, as defined by Chemical Abstracts nomenclature is shown below.
The compounds of Formula (I-a) and (I-b) have at least two asymmetric centers at respectively carbon atom 2 and 3. Said asymmetric center and any other asymmetric center, which may be present (e.g. at atom 8 in (I-a) or 9 in (I-b)), are indicated by the descriptors R and S. When e.g. a monocyanomethylene moiety is present in the compounds of Formula (I-a) at position 8, said moiety may have the E- or Z-configuration.
The invention also comprises derivative compounds (usually called “pro-drugs”) of the pharmacologically active compounds according to the invention, which are degraded in vivo to yield the compounds according to the invention. Pro-drugs are usually (but not always) of lower potency at the target receptor than the compounds to which they are degraded. Pro-drugs are particularly useful when the desired compound has chemical or physical properties that make its administration difficult or inefficient. For example, the desired compound may be only poorly soluble, it may be poorly transported across the mucosal epithelium, or it may have an undesirably short plasma half-life. Further discussion on pro-drugs may be found in Stella, V. J. et al., “Prodrugs”, Drug Delivery Systems, 1985, pp. 112-176, and Drugs, 1985, 29, pp. 455-473.
Prodrugs forms of the pharmacologically-active compounds according to the invention will generally be compounds according to Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof and the N-oxide form thereof, having an acid group which is esterified or amidated. Included in such esterified acid groups are groups of the Formula —COORx, where Rx is a C1-6alkyl, phenyl, benzyl or one of the following groups:
Amidated groups include groups of the Formula —CONRyRz, wherein Ry is H, C1-6alkyl, phenyl or benzyl and Rz is —OH, H, C1-6alkyl, phenyl or benzyl. Compounds according to the invention having an amino group may be derivatised with a ketone or an aldehyde such as formaldehyde to form a Mannich base. This base will hydrolyze with first order kinetics in aqueous solution.
The compounds of Formula (I) as prepared in the processes described below may be synthesized in the form of racemic mixtures of enantiomers that can be separated from one another following art-known resolution procedures. The racemic compounds of Formula (I) may be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. Said diastereomeric salt forms are subsequently separated, for example, by selective or fractional crystallization and the enantiomers are liberated therefrom by alkali. An alternative manner of separating the enantiomeric forms of the compounds of Formula (I) involves liquid chromatography using a chiral stationary phase. Said pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifically. Preferably if a specific stereoisomer is desired, said compound would be synthesized by stereospecific methods of preparation. These methods will advantageously employ enantiomerically pure starting materials.
Pharmacology
The compounds of the present invention show affinity for 5-HT2 receptors, particularly for 5-HT2A and 5-HT2C receptors (nomenclature as described by D. Hoyer in “Serotonin (5-HT) in neurologic and psychiatric disorders” edited by M. D. Ferrari and published in 1994 by the Boerhaave Commission of the University of Leiden) and affinity for the D2 receptor as well as norepinephrine reuptake inhibition activity. The serotonin antagonistic properties of the present compounds may be demonstrated by their inhibitory effect in the “5-hydroxytryptophan Test on Rats” which is described in Drug Dev. Res., 13, 237-244 (1988).
In view of their capability to block 5-HT2 receptors, and in particular to block 5-HT2A and 5-HT2C receptors, as well as the D2 receptor and by also effecting the norepinephrine reuptake inhibition activity, the compounds according to the invention are useful as a medicine, in particular in the prophylactic and therapeutic treatment of conditions mediated through either of these receptors.
The invention therefore relates to a compound according to the general Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form thereof and prodrugs thereof, for use as a medicine.
The invention also relates to the use of a compound according to the general Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form thereof and prodrugs thereof for the manufacture of a medicament for treating, either prophylactic or therapeutic or both, conditions mediated through the 5-HT2, and D2 receptor, as well as the through norepinephrine reuptake inhibition.
In view of these pharmacological and physicochemical properties, the compounds of Formula (I) are useful as therapeutic agents in the treatment or the prevention of central nervous system disorders like anxiety, depression and mild depression, bipolar disorders, sleep- and sexual disorders, psychosis, borderline psychosis, schizophrenia, migraine, personality disorders or obsessive-compulsive disorders, social phobias or panic attacks, organic mental disorders, mental disorders in children such as ADHD, aggression, memory disorders and attitude disorders in older people, addiction, obesity, bulimia and similar disorders. In particular, the present compounds may be used as anxiolytics, antidepressants, antipsychotics, anti-schizophrenia agents, anti-migraine agents and as agents having the potential to overrule the addictive properties of drugs of abuse.
The compounds of Formula (I) may also be used as therapeutic agents in the treatment of motoric disorders. It may be advantageous to use the present compounds in combination with classical therapeutic agents for such disorders.
The compounds of Formula (I) may also serve in the treatment or the prevention of damage to the nervous system caused by trauma, stroke, neurodegenerative illnesses and the like; cardiovascular disorders like high blood pressure, thrombosis, stroke, and the like; and gastrointestinal disorders like dysfunction of the motility of the gastrointestinal system and the like.
In view of the above uses of the compounds of Formula (I), it follows that the present invention also provides a method of treating warm-blooded animals suffering from such diseases, said method comprising the systemic administration of a therapeutic amount of a compound of Formula (I) effective in treating the above described disorders, in particular, in treating anxiety, psychosis, depression, migraine and addictive properties of drugs of abuse.
The present invention thus also relates to compounds of Formula (I) as defined hereinabove for use as a medicine, in particular, the compounds of Formula (I) may be used for the manufacture of a medicament for treating anxiety, psychosis, depression, migraine and addictive properties of drugs of abuse.
Those of skill in the treatment of such diseases could determine the effective therapeutic daily amount from the test results presented hereinafter. An effective therapeutic daily amount would be from about 0.01 mg/kg to about 10 mg/kg body weight, more preferably from about 0.05 mg/kg to about 1 mg/kg body weight.
The invention also relates to a pharmaceutical composition comprising a pharmaceutically acceptable carrier and, as active ingredient, a therapeutically effective amount of a compound according to the invention, in particular a compound according to Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form thereof and a prodrug thereof.
The compounds according to the invention, in particular the compounds according to Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form thereof and the prodrugs thereof, or any subgroup or combination thereof may be Formulated into various pharmaceutical forms for administration purposes. As appropriate compositions there may be cited all compositions usually employed for systemically administering drugs. To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, optionally in addition salt form, as the active ingredient is combined in intimate admixture with a pharmaceutically acceptable carrier, which carrier may take a wide variety of forms depending on the form of preparation desired for administration. These pharmaceutical compositions are desirable in unitary dosage form suitable, in particular, for administration orally, rectally, percutaneously, by parenteral injection or by inhalation. For example, in preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed such as, for example, water, glycols, oils, alcohols and the like in the case of oral liquid preparations such as suspensions, syrups, elixirs, emulsions and solutions; or solid carriers such as starches, sugars, kaolin, diluents, lubricants, binders, disintegrating agents and the like in the case of powders, pills, capsules and tablets. Because of their ease in administration, tablets and capsules represent the most advantageous oral dosage unit forms in which case solid pharmaceutical carriers are obviously employed. For parenteral compositions, the carrier will usually comprise sterile water, at least in large part, though other ingredients, for example, to aid solubility, may be included. Injectable solutions, for example, may be prepared in which the carrier comprises saline solution, glucose solution or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be employed. Also included are solid form preparations that are intended to be converted, shortly before use, to liquid form preparations. In the compositions suitable for percutaneous administration, the carrier optionally comprises a penetration enhancing agent and/or a suitable wetting agent, optionally combined with suitable additives of any nature in minor proportions, which additives do not introduce a significant deleterious effect on the skin. Said additives may facilitate the administration to the skin and/or may be helpful for preparing the desired compositions. These compositions may be administered in various ways, e.g., as a transdermal patch, as a spot-on, as an ointment.
It is especially advantageous to Formulate the aforementioned pharmaceutical compositions in unit dosage form for ease of administration and uniformity of dosage. Unit dosage form as used herein refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including scored or coated tablets), capsules, pills, powder packets, wafers, suppositories, injectable solutions or suspensions and the like, and segregated multiples thereof.
Since the compounds according to the invention are potent orally administrable compounds, pharmaceutical compositions comprising said compounds for administration orally are especially advantageous.
In order to enhance the solubility and/or the stability of the compounds of Formula (I) in pharmaceutical compositions, it can be advantageous to employ α-, β-or γ-cyclodextrins or their derivatives, in particular hydroxyalkyl substituted cyclodextrins, e.g. 2-hydroxypropyl-β-cyclodextrin. Also co-solvents such as alcohols may improve the solubility and/or the stability of the compounds according to the invention in pharmaceutical compositions.
Preparation
Suitable preparation schemes for the compounds of the invention are described below:
The following abbreviations are used thoughout the text:
The following reaction schemes A to D illustrate the preparation of compounds of formula (I) in which C is a group of formula (c-1) in which Y1 is NH and R11 is a group of formula (d-1), represented by formulae Ia and Ib below:
An intermediate compound 10 leads to a final compound of formula (Ia); an intermediate compound 13 leads to a final compound of formula (Ib).
Methods B-D below represent alternative routes to the preparation of the above final compounds of formula Ia and Ib:
Method B: Synthesis of (2R,3aR,12bS)- and (2S,3aR,12bS)-Final Compounds
The following reaction scheme illustrates the preparation of compounds of formula (I) in which C is a group of formula (c-1) in which R11 and R10 form a condensed imidazole residue, represented by formula II below.
The following reaction schemes illustrate the preparation of compounds of formula (I) in which C is a group of formula (c-1) in which R11 and R10 form a condensed piperazine residue, represented by formula III below in which Rx is hydrogen or alkyl and the piperazine ring has the S configuration (Scheme F1) or the R configuration (Scheme F2):
The following reaction scheme G illustrates the preparation of compounds of formula (I) in which C is a group of formula (c-1) and X is a CR6R7 group other than a hydrogen group, represented by formulae IV-VI below.
The following reaction scheme H illustrates the preparation of compounds of formula (I) in which C is a group of formula (c-3), Y1 is NH and R11 is a group of formula (d-1) represented by formula VII below.
The following reaction scheme I illustrates the preparation of compounds of formula (I) in which C is a group of formula (c-3), Y2 is O and R11 is a group of formula (d-1), represented by formula VIII below.
The following reaction scheme J illustrates the preparation of compounds of formula (I) in which C is a group of formula (c-2), Y2 is O and R11 is a group of formula (d-1), represented by formula IX below. The compound can be either cis (Scheme J1) or trans (Scheme J2) with respect to the oxygen.
The reaction scheme J1 can also be applied to the trans epimer of intermediate compound 52, leading to the trans-compounds (IX) and (X).
Method K: Preparation of 4-Substituted Tetrahydropyran-Derivatives
The following reaction scheme K illustrates the preparation of compounds of formula (I) in which C is a group of formula (c-2), Y2 is O and R11 is a group of formula (d-1), represented by formula X below.
The following reaction schemes L1-L3 illustrates the preparation of compounds of formula (I) in which C is a group of formula (c-1), Y1 is SO(n) and R11 is a group of formula (d-1), represented by formulae XIa-c and XIIa-c below.
The compounds of Formula (I) may also be converted into each other following art-known transformation reactions. For instance,
The procedures described above can be modified by the use of conventional procedures which will be known to those skilled in the art to provide analogous processes for the preparation of compounds of formula (I).
The starting materials mentioned hereinabove are either commercially available or may be made following art-known procedures. For instance, intermediates 1 may be prepared in accordance with the techniques described in patent specifications WO 03/048146 and WO03/048147 referred to above or by techniques analogous thereto.
Pure stereochemically isomeric forms of the compounds of Formula (I) may be obtained by the application of art-known procedures. Diastereomers may be separated by physical methods such as selective crystallization and chromatographic techniques, e.g. counter-current distribution, liquid chromatography and the like.
The compounds of Formula (I) as prepared in the hereinabove described processes are generally racemic mixtures of enantiomers which can be separated from one another following art-known resolution procedures. The racemic compounds of Formula (I) which are sufficiently basic or acidic may be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid respectively with a suitable chiral base. Said diastereomeric salt forms are subsequently separated, for example, by selective or fractional crystallization and the enantiomers are liberated therefrom by alkali or acid. An alternative manner of separating the enantiomeric forms of the compounds of Formula (I) involves liquid chromatography using a chiral stationary phase. Said pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifically. Preferably if a specific stereoisomer is desired, said compound will be synthesized by stereospecific methods of preparation. These methods will advantageously employ enantiomerically pure starting materials.
The following examples are intended to illustrate and not to limit the scope of the present invention.
Experimental Part
A. Preparation of the Intermediate Compounds
A solution of α,β-unsaturated ketone intermediate 1 (1.00 g, 2.96 mmol) and Et3N (0.63 mL, 4.50 mmol) in i-PrOH (30 mL) was hydrogenated with 10% Pd/C at atmospheric pressure for 6 hour. Then the mixture was filtered through a pad of celite and the solids were washed with CH2Cl2 (4×20 mL). After evaporation, i-PrOH (5 mL) and Et3N (1.20 mL) was added and the reaction mixture was stirred at 40° C. for 1 hour. The reaction mixture was cooled to room temperature and allowed to crystallize. The crystals were filtered off and dried under vacuum to afford pure ketone intermediate 2 as a white crystalline powder (0.86 g, 86%); mp: 144-146° C.
Mass spectrum: CI m/z (assignment, relative intensity) 341 (MH+, 2%), 283 (MH+-acetone, 100%); EI: m/z (assignment, relative intensity) 340 (M+., 1%), 282 (M−. -acetone, 79%), 226 (M+.-sidechain+H, 100%); High resolution EI, Calculated C21H21FO3 (M+.): 340.1475, Found: 340.1479 (1%).
To an ice-cooled solution of ketone intermediate 2 (0.42 g, 1.23 mmol) in i-PrOH (15 mL) was added phosphate buffer solution (pH=7, 5 mL) and then portionwise NaBH4 (0.23 g, 6.16 mmol). The reaction mixture was stirred at room temperature for 1 hour. Then 10 mL NH4Cl (sat. aq. solution) was added, the mixture was extracted with CH2Cl2 (3×15 mL) and the organic phases were dried with MgSO4. After removal of the solvent, the residue was purified on a silica gel column by using ether/hexane (40:60), yielding intermediate 3 as a colorless oil (0.42 g, 99%).
Mass spectrum: CI m/z (assignment, relative intensity) 325 (MH+—H2O, 53%), 267 (MH+—H2O-acetone, 100%), 249 (MH+—2H2O-acetone, 97%); EI: m/z (assignment, relative intensity) 342 (M+., 3%), 324 (M+.—H2O, 48%), 266 (M+.—H2O-acetone, 35%), 209 (100%); High resolution EI Calculated C21H23FO3 (M+.): 342.1631, Found: 342.1627 (5%).
To a cooled (−30° C.) solution of DIAD (2.43 mL, 33.47 mmol) in THF (10 mL) were added intermediate alcohol 3 (2.30 g, 6.73 mmol) in THF (18 mL) and Ph3P (3.71 g, 14.07 mmol). After 20 minutes, diphenyl phosphoryl azide (DPPA) (3.62 mL, 16.83 mmol) was added and the reaction mixture was allowed to warm up to room temperature. After stirring overnight, the solvent was removed in vacuo to give a red oil. The crude material was purified by column chromatography using ether/hexane (10/90) to give an unseparated mixture of intermediate 4, as an oil, and Ph3PO (3.46 g).
Mass spectrum: CI m/z (assignment, relative intensity) 368 (MH+, 1%), 325 (MH+—HN3, 9%), 304 (13%), 276 (MH+—HN3-acetone, 100%), 248 (20%).
To solution of azide intermediate 4 (3.68 g, 10.02 mmol) in THF (30 mL) was added 1N HCl (30 mL) and the mixture was stirred at room temperature for 8 hours. Add K2CO3 (sat. aq. sol.) at 0° C., extract 3 times with CH2Cl2 and dry with MgSO4. The residue obtained upon evaporation was purified by column chromatography on silica gel using Et2O/heptane (30/70) to give an oily intermediate 5 (3.19 g, 91% for 2 steps from 3).
Mass spectrum: CI m/z (assignment, relative intensity) 328 (MH+, 2%), 310 (MH+—H2O, 2%), 300 (MH+—N2, 5%), 285 (MH+—HN3, 11%), 267 (MH+—HN3—H2O, 100%), 249 (MH+—HN3—2H2O, 33%), 225 (MH+—HN3—CH2OHCHO, 20%).
To solution of diol intermediate 5 (1.11 g, 3.39 mmol) in dry toluene (10 mL) was added Bu2SnO (97.6 mg, 0.39 mmol), Et3N (1.07 mL, 7.74 mmol) and TsCl (0.739 g, 3.87 mmol). The mixture was stirred at room temperature overnight. Add NH4Cl (sat. aq. sol.), extract 3 times with CH2Cl2 and dry with MgSO4. The residue was purified by column chromatography on silica gel using EtOAc/heptane (20/80) to give intermediate 6 as an oil (1.55 g, 95%).
Mass spectrum: —CI m/z (assignment, relative intensity) 454 (MH+—N2, 1%), 421 (MH+—HN3—H2O, 1%), 282 (MH+—TsOH—HN3, 20%), 264 (MH+—TsOH—HN3—H2O, 15%), 173 (TsOH2+, 100%).
A solution of tosylate intermediate 6 (2.00 g, 4.15 mmol) in DMF (30 mL) was treated with sodium azide (810.8 mg, 12.47 mmol) and the mixture was stirred at 90° C. in the dark for 2 hours. The reaction mixture was diluted with water and extracted with CH2Cl2. The combined extracts were washed with brine. Following concentration of the dried organic phases the residue was purified by column chromatography on silica gel using heptane/EtOAc (80/20) affording diazide intermediate 7 (1.22 g, 88%) as an oil.
Mass spectrum: —CI m/z (assignment, relative intensity) 325 (MH+—N2, 2%), 310 (MH+—HN3, 3%), 297 (MH+—N2—N2, 1%), 282 (MH+—HN3—N2, 52%), 268 (MH+—HN3—HN3, 3%).
To solution of diazide intermediate 7 (65 mg, 0.18 mmol) in CH2Cl2 (10 mL) was added DMAP (18.5 mg, 0.09 mmol), Et3N (0.13 mL, 0.63 mmol) and MsCl (44.5 μL, 0.40 mmol). After stirring at room temperature for 10 minutes, 10 mL NH4Cl (sat. aq. solution) was added. Extract with CH2Cl2 (3×10 mL) and dry with MgSO4. Column purification on silica gel using EtOAc/heptane (20:80) yielded intermediate 8 as an oil (78.2 mg, 98%).
Mass spectrum: —CI m/z (assignment, relative intensity) 403 (MH+—N2, 3%), 360 (MH+ —N2—HN3, 43%), 307 (MH+-MeSO3H—N2, 50%), 264 (MH+-MeSO3H—HN3—N2, 58%), 250 (MH+-MeSO3H—HN3, —N3, 21%), 197 (100%).
A solution of diazide intermediate 8 (98.2 mg, 0.23 mmol) in MeOH (10 mL) was hydrogenated at atmospheric pressure with 10% Pd/C for 1 night. Then the mixture was filtered through a pad of celite and the solids were washed 4 times with CH2Cl2. After evaporation of the filtrate, the crude product was purified by column chromatography on silica gel using CHCl3/MeOH/NH4OH (90/9/1). This afforded intermediate 9 as an oil (36.4 mg, 56%).
To a cooled (0° C.) solution of DIAD (4.2 mL, 21.18 mmol) in THF (50 mL) was added Ph3P (5.55 g, 21.18 mmol). Stir at 0° C. for 30 minutes (precipitation of white solid). Then, a mixture of alcohol intermediate 7 (3.727 g, 10.59 mmol) and 4-nitrobenzoic acid (3.54 g, 21.18 mmol) in THF (50 mL) was added. The reaction mixture was allowed to warm up to room temperature and after stirring for 2 hours, MeOH was added and the stirring continued for an additional 30 minutes. After removal of the solvent, the crude material was purified by column chromatography using EtOAc/heptane (20/80) to give the ester intermediate 10 as an oil (4.85 g, 91%).
Mass spectrum: —CI m/z (assignment, relative intensity) 431 (MH+—N2—HN3, 36%), 307 (MH+—N2-p-NO2PHCO2H, 2%), 264 (MH+-p-NO2PHCO2H—HN3—N2, 58%), 197 (100%), 182 (72%).
A solution of the above diazide intermediate 10 (78.0 mg, 0.15 mmol) in MeOH (2 mL) was treated with K2CO3 (76.9 mg, 0.47 mmol) and the mixture was stirred for 1 hour. Add NH4Cl (sat. aq. sol.), extract 3 times with CH2Cl2 and dry with MgSO4. The residue was purified by column chromatography on silica gel using EtOAc/heptane (20/80) to give alcohol intermediate 11 as an oil (42.6 mg, 78%).
Mass spectrum: —CI m/z (assignment, relative intensity) 325 (MH+—N2, 2%), 310 (MH+ —HN3, 3%), 297 (MH+—N2—N2, 1%), 282 (MH+—HN3—N2, 52%), 268 (MH+—NH3—N3, 3%).
To a solution of diazide intermediate 11 (42.6 mg, 0.12 mmol) in CH2Cl2 (5 mL) was added DMAP (12.7 mg, 0.06 mmol), Et3N (0.047 mL, 0.42 mmol) and MsCl (33.9 μL, 0.30 mmol). Stir at room temperature for 10 minutes. Add 10 mL NH4Cl (sat. aq. solution), extract with CH2Cl2 (3×10 mL) and dry with MgSO4; upon evaporation of the solvent intermediate 12 was obtained as an oil (53.0 mg, 100%).
Mass spectrum: —CI m/z (assignment, relative intensity) 403 (MH+—N2, 3%), 360 (MH+ —N2—HN3, 43%), 307 (MH+-MeSO3H—N2, 50%), 264 (MH+-MeSO3H—HN3—N2, 58%), 250 (MH+-MeSO3H—HN3—N3, 21%), 197 (100%).
A solution of diazide intermediate 12 (501.0 mg, 1.16 mmol) in MeOH (10 mL) was hydrogenated under 1 atmospheric pressure with 10% palladium-on-charcoal under vigorous stirring at room temperature for 1 night. Then the mixture was filtered through a pad of celite and the solids were washed 4 times with CH2Cl2. After evaporation, the crude product was purified by column chromatograhy on silica gel using CHCl3/MeOH/NH4OH (90/9/1). This yielded intermediate 13 as an oil (270.0 mg, 82%).
To a solution of diamine intermediate 9 (220.0 mg, 0.78 mmol) in CH2Cl2 (5 mL) at −20° C. was added Et3N (0.109 mL, 0.78 mmol) and benzyl chloroformate (0.112 mL, 0.78 mmol). The mixture was then stirred for 1 hour. Add 10 mL of NH4Cl (sat. aq. solution), extract with CH2Cl2 (3×10 mL) and dry with MgSO4. The residue was purified by column chromatography on silica gel using EtOAc/heptane (20/80) to give a mono-Cbz intermediate 14 (128.9 mg, 40%) and di-Cbz derivative (84.5 mg).
Mass spectrum: —CI m/z (assignment, relative intensity) 417 (MH+, 100%), 397 (MH+—HF, 8%), 311 (MH+-PhCHO, 7%), 309 (MH+—PHCH2OH, 32%), 283 (16%), 252 (MH+-PhCH2OCONHCH3, 24%).
To a solution of monoCbz intermediate 14 (32.5 mg, 0.078 mmol) in EtOAc (3 mL) was added 1 mL of NaOH (sat. aq. solution) and bromoacetyl bromide (6.8 μL, 0.078 mmol). The two phases were stirred vigorously for 1 night. Add 10 mL of NH4Cl (sat. aq. solution), extract with CH2Cl2 (3×10 mL) and dry with MgSO4. Column purification on silica gel using EtOAc/heptane (20/80) gave intermediate 15 as an oil (31.4 mg, 62%).
Mass spectrum: —CI m/z (assignment, relative intensity) 457 (MH+—HBr, 3%), 413 (MH —HBr—CO2, 1%), 365 (MH+—HBr—PhCH3 1%), 351 (MH+-PhCHO—HBr, 2%), 323 (MH+—HBr—PhCHO—CO, 5%), 119 (8%), 91 (100%).
To a solution of the above carbamate intermediate 15 (91.7 mg, 0.17 mmol) in DMF (5 mL) was added K2CO3 (103.0 mg, 0.75 mmol) and the mixture was stirred at room temperature for 36 hours. Add 10 mL of NH4Cl (sat. aq. solution), extract with CH2Cl2 (3×10 mL) and dry with MgSO4. Column purification on silica gel using EtOAc/heptane (30/70) gave polycyclic intermediate 16 (86.2 mg, 92%) as an oil.
Mass spectrum: —CI m/z (assignment, relative intensity) 457 (MH+, 1%), 323 (MH+—PhCHO—CO, 5%), 279 (MH+-Cbz-CH2CO, 1%), 91 (10%).
To an ice cooled solution of diamine 13 (41.6 mg, 0.15 mmol) in CH2Cl2 (5 mL) was added Et3N (42.5 μL, 0.3 mmol), DMAP (9.4 mg, 0.07 mmol) and trityl chloride (46.1 mg, 0.16 mmol). The mixture was then stirred at 0° C. for 2 hours. Add 10 mL of NH4Cl (sat. aq. solution), extract with CH2Cl2 (3×10 mL) and dry with MgSO4. Column purification on silica gel using EtOAc/heptane (20/80) gave a crystalline intermediate 17 (52.6 mg, 68%); mp: 58-60° C.
Mass spectrum: -APCI m/z (assignment, relative intensity) 525 (MH+, 38%), 390 (4%), 283 (MH+-(Tr-H), 15%), 252 (MH+—CH3NHTr, 27%), 243 (Tr+, 100%), 228 (7%), 165 (29%).
The diamine intermediate 17 (26.7 mg, 0.05 mmol) was added to a two-phase system consisting of 2 mL CH2Cl2 and 0.5 mL Na2CO3 (aq. sat. solution), and the mixture was stirred for 10 minutes. After adding bromoacetyl bromide (6.8 μL, 0.08 mmol) the two phases were stirred vigorously for 3 hours. Extract with CH2Cl2 (3×10 mL) and dry with MgSO4. Column purification on silica gel using EtOAc/heptane (20/80) gave intermediate 18 as an oil (27.9 mg, 85%) characterised as a mixture of two conformers.
Mass spectrum: -APCI m/z (assignment, relative intensity) 645 (MH+, 39%), 601 (3%), 403 (MH+-(Tr-H), 7%), 321 (MH+—TrH—HBr, 21%), 243 (Tr+, 100%), 228 (3%), 165 (15%).
To a solution of intermediate 18 (530 mg, 0.82 mmol) in MeOH (15 mL) was added MeSO3H (3 mL) and the mixture was stirred at 60° C. for 30 minutes. After complete evaporation of the solvent, the residue was dissolved in CH2Cl2/K2CO3 (sat. aq. solution) (15/15 mL) and the organic layer was separated. The aqueous layer was extracted with CH2Cl2 (3×10 mL) and the combined organic layers were then dried with MgSO4. Column purification on silica gel using EtOAc/heptane (20/80) gave intermediate 19 as an oil (231.3 mg, 47%), characterised as a mixture of two conformers.
Mass spectrum: -APCI m/z (assignment, relative intensity) 598 (MH+, 1%), 519 (2%), 355 (MH+-Tr, 13%), 283 (MH+-Tr-CO═CHOMe, 2%), 271 (10%), 243 (Tr+, 100%), 167 (21%).
The N-Tr protected amine intermediate 18 (100 mg, 0.15 mmol) was dissolved in 98% formic acid (2 mL) and the mixture was stirred at room temperature for 24 hours. After removal of excess of formic acid in vacuo, the residue was dissolved in CHCl3 (2 mL) and EEDQ (47 mg, 0.19 mmol) was added. The solution was stirred at room temperature for 5 hours. Following evaporation of the solvent, the residue was purified by column chromatography on silica gel using CH2Cl2/MeOH (98/2) as eluent. The N—CHO protected amine intermediate 20 (54.7 mg, 82%) was obtained as an oil.
Mass spectrum: —CI m/z (assignment, relative intensity) 431, 433 (MH+, 42%), 353 (MH+—HBr, 100%), 294 (MH+—HBr—CH3NHCHO, 9%), 249 (4%), 158 (2%), 130 (7%).
To a solution of N-formyl protected amine intermediate 20 (91 mg, 0.21 mmol) in dry THF (10 mL) was added a solution of t-BuOK (30.3 mg, 0.24 mmol) in THF (2 mL). The reaction mixture was stirred at room temperature for 30 minutes. Water (10 mL) was then added and the mixture extracted with CH2Cl2 (10 mL). Column purification on silica gel using CH2Cl2/MeOH (97/3) gave the cyclic intermediate 21 (47.4 mg, 64%) as an oil.
Mass spectrum: —CI m/z (assignment, relative intensity) 351 (MH+, 100%), 331 (MH+—HF, 5%), 323 (MH+—CO, 6%), 319 (8%), 219 (2%), 130 (4%).
To a solution of the alcohol intermediate 3 (0.42 g, 1.23 mmol) in CH2Cl2 (30 mL) was added Et3N (0.43 mL, 3.07 mmol), DMAP (0.15 g, 1.23 mmol) and AcOH anhydride (0.29 mL, 3.07 mmol). Stir at room temperature for 1 hour, add NH4Cl (sat. aq. solution, 20 mL), extract with CH2Cl2 (3×15 mL) and dry with MgSO4. Column purification on silica gel using ether/hexane (30:70) gave a white crystalline intermediate 22 (0.45 g, 95%); mp: 147-149° C.
Mass spectrum: —CI m/z (assignment, relative intensity) 385 (MH+, 1%), 325 (MH+—AcOH, 100%), 267 (MH+—AcOH-acetone, 43%), 249 (MH+—AcOH-acetone-H2O, 47%); -EI: m/z (assignment, relative intensity) 324 (M+.—AcOH, 46%), 266 (M+.—AcOH-acetone, 20%), 209(M+.—AcOH-sidechain, 100%); High resolution EI Calculated C22H21FO2 (M+.—AcOH): 324.1526, Found: 324.1521 (M+., 72%).
To a solution of the acetal intermediate 22 (0.45 g, 1.17 mmol) in THF (10 mL) was added 1N HCl (10 mL). After stirring at room temperature for 8 hours, 10 mL Na2CO3 (sat. aq. solution) was added at 0° C. Extract with CH2Cl2 (3×10 mL) and dry with MgSO4. Column purification on silica gel using EtOAc/hexane (70:30) gave diol intermediate 23 as a colorless oil (0.39 g, 96%).
Mass spectrum: —CI m/z (assignment, relative intensity) 345 (MH+, 1%), 327 (MH+—H2O, 3%), 309 (MH+—2 H2O, 3%), 285 (MH+—AcOH, 17%), 267 (MH+—AcOH—H2O, 100%), 249 (MH−—AcOH-2 H2O, 3%); EI: m/z (assignment, relative intensity) 326 (M+.—H2O, 10%), 284 (M+.—AcOH, 13%), 209 (M+.—AcOH-sidechain, 100%)); High resolution EI Calculated C20H19FO3 (M+.—H2O): 326.1318, Found: 326.1316 (31%).
To the acetate diol intermediate 23 (0.59 g, 1.72 mmol) in CH2Cl2 (15 mL) was added Et3N (0.96 mL, 6.86 mmol), DMAP (209 mg, 1.72 mmol) and MeSO2Cl (0.53 mL, 6.86 mmol) at 0° C. Stir at room temperature for 1 hour. Work it up by adding NH4Cl (sat. aq. sol.), extract 3 times with CH2Cl2 and dry with MgSO4. Column purification on silica gel using EtOAc/heptane (50/50) afforded dimesyl compound as an oil (0.84 g, 98%). To this compound (182.5 mg, 0.36 mmol) in DMF (10 mL) was added NaN3 (95 mg, 1.46 mmol). The reaction mixture was heated at 80° C. for 3 hours. After cooling, add NH4Cl (sat. aq. sol.), extract 3 times with CH2Cl2 and dry with MgSO4. After evaporation the residue was purified on silica gel using EtOAc/heptane (20/80) to give intermediate 24 as an oily product (122.3 mg, 85%).
Diazide intermediate 24 was converted via diazido alcohol intermediate 24a into a diamine which was further converted into intermediate 25. To a solution of diazide 24 (120.1 mg, 0.30 mmol) in MeOH (10 mL) was added K2CO3 (126.4 mg, 0.91 mmol). The reaction mixture was stirred at room temperature for 1 hour. Add NH4Cl (sat. aq. sol.), extract 3 times with CH2Cl2. Column purification on silica gel using Et2O/heptane (40/60) gave the diazido alcohol intermediate 24a as an oily product (77.5 mg, 72%). This compound (75 mg, 0.21 mmol) in MeOH (5 mL) was hydrogenated at 1 atmospheric pressure with 10% palladium-on-charcoal under vigorous stirring at room temperature for 1 night. Then the mixture was filtered through a pad of celite and the solids were washed 4 times with CH2Cl2. After evaporation of the solvent, the crude product was dissolved in 5 mL of CH3CN and Et3N (34 μL, 0.24 mmol) was added. The reaction mixture was heated under argon at 70° C. After 1 hour, a solution of diphenyl carbonate (23 mg, 0.11 mmol) in CH3CN was added dropwise and the mixture was stirred at 70° C. for 1 day. After evaporation, the crude product was purified by column chromatography on silica gel using CHCl3/MeOH (90/10) to give the imidazolidinone intermediate 25 as an oil (34.4 mg, 48%).
To a suspension of intermediate 26 (0.228 g, 1 mmol) and MgBr2 (0.202 g, 1.1 mmol) in dry toluene (5 mL), (S)-glyceraldehyde acetonide (4 mmol, 1.5 M solution in THF) and tBuOK (22.4 mg, 0.2 mmol) was added and stirred for 3 hours at room temperature. A saturated aq. NH4Cl solution (5 mL) was added, the organic layer was separated and kept over anhydrous MgSO4. The solvent was removed under reduced pressure followed by the separation of α,β-unsaturated product by flash chromatography using EtOAc:heptane (1:9) as an eluant to obtain intermediate 27 as a yellow liquid in a ratio of 85/15 E and Z isomer (85%, 0.289 g).
HRMS: Calculated 340.1111; found 340.1122
To a solution of intermediate 27 (0.340 g, 1 mmol) in i-PrOH (5 mL) was added Et3N (0.21 mL, 1.5 mmol) and the reaction mixture was hydrogenated under atmospheric pressure using 10% Pd/C (40 mg) as a catalyst. After completion of the reaction (4 hours) the reaction mixture was passed through a small pad of celite and further washed with CH2Cl2 (2×5 mL) followed by the evaporation of the solvent to obtain crude ketone intermediate 28.
b) The crude intermediate 28 thus obtained was dissolved in i-PrOH (10 mL) and aqueous phosphate buffer solution (3 mL, pH 7) was added to it. The temperature was lowered to 0° C. and NaBH4 (0.152 g, 4 mmol) was added to it in several lots and then allowed to stir further for 15 minutes at the same temperature. Aq. NH4Cl solution (5 mL) was added and the reaction mixture was extracted using Et2O (3×5 mL). After drying over anhydrous MgSO4 the solvent was removed under reduced pressure and the two diastereomeric alcohols (1:1) with slightly different polarity were separated by flash chromatography using EtOAc:heptane (20:80) as an eluant to obtain the more polar cis-alcohol intermediate 29 as a white solid (mp: 59-61° C.; 49%, 0.16 g).
HRMS: Calculated 344.1424; found 344.1435
To a solution of P(Ph)3 (0.524 g, 2 mmol) in dry THF (5 mL) at −15° C., a solution of DIAD (0.424 g, 2.1 mmol) in THF (2 mL) was added and the resulting complex was stirred for 20 minutes followed by the addition of intermediate 29 (0.329 g, 1 mmol) dissolved in THF (2 mL) and a solution of DPPA (0.330 g, 1.2 mmol) in THF (1 mL). The reaction mixture was warmed to room temperature and stirred for 18 hours. After addition of MeOH the reaction mixture was dried under vacuum followed by separation of the azide using flash chromatography employing EtOAc:heptane (1:9) as an eluant to obtain intermediate 30 as a colourless liquid (91%, 0.335 g).
HRMS: Calculated 369.1489; found 369.1483.
To a solution of intermediate 30 (0.369 g, 1 mmol) in THF (5 mL) 1M aq. HCl solution (1 mL) was added and stirred for 18 hours. THF was removed under reduced pressure and the diol was extracted using Et2O (3×10 mL). The organic layer was treated with aq. NaHCO3 (5 mL) followed by a brine wash (5 mL). After drying over anhydrous MgSO4 the solvent was removed under vacuum to obtain intermediate 31 as a thick viscous liquid (95%, 0.313 g).
HRMS: Calculated 329.1176; found 329.1184.
To a solution of intermediate 31 (0.329 g, 1 mmol) in CH2Cl2 (10 mL) Et3N (0.28 mL, 2 mmol), DMAP (0.1 mmol, 12.2 mg) and TrCl (0.307 g, 1.1 mmol) were added and stirred for 24 hours. The solvent was removed under reduced pressure and the crude reaction mixture was subjected to flash column chromatography using EtOAc:heptane (1:9) as an eluant to obtain intermediate 32 as a white solid (mp: 58-59° C.; 80%, 0.456 g).
HRMS: Calculated 571.2271; found 571.2286.
To a solution of intermediate 32 (0.571 g, 1 mmol) in CH2Cl2 at −10° C., Et3N (0.28 mL, 2 mmol), DMAP (12.2 mg, 0.1 mmol) and MsCl (0.126 g, 1.1 mmol) were added. The reaction mixture was warmed up to room temperature and stirred for 4 hours. Water (3 mL) was added and the organic layer was separated and dried over anhydrous MgSO4 followed by the purification by flash chromatography using EtOAc:heptane (1:9) as an eluant to obtain intermediate 33 as a white solid (mp:55-56° C.; 85%, 0.515 g).
HRMS: Calculated 649.2047; found 649.2064
To a solution of intermediate 33 (0.649 g, 1 mmol) in MeOH (5 mL), amberlyst-15 (0.1 g) was added and the reaction mixture was stirred at 40° C. for 3 hours, then filtered to remove the catalyst. The solvent was removed under reduced pressure and the product purified by flash chromatography using EtOAc:heptane (2:8) as an eluant to obtain intermediate 34 as a thick viscous liquid (90%, 0.366 g).
HRMS: Calculated 407.0951; found 407.0975.
A mixture of intermediate 34 (0.407 g, 1 mmol) and K2CO3 (0.276 g, 2 mmol) was stirred in i-PrOH (10 mL) for 8 hours, filtered to remove K2CO3 and the solvent was removed under reduced pressure. The product was purified by flash chromatograpy using EtOAc:heptane (2:8) as an eluant to obtain intermediate 35 as a colourless liquid (78%, 0.242 g).
HRMS: Calculated 311.1070; found 311.1089.
To a solution of intermediate 35 (0.311 g, 1 mmol) in i-PrOH (10 mL), Et3N (0.140 mL, 1 mmol) was added. The mixture was hydrogenated under atmospheric pressure using 10% Pd/C (50 mg) as a catalyst. After completion of the reaction (3 hours), it was passed through a small pad of celite and the catalyst was washed with CH2Cl2 (2×5 mL). The combined organic layers were evaporated under reduced pressure and purified by flash chromatography using EtOAC:heptane (1:1) as an eluant to obtain intermediate 36 as a white solid (mp: 108-109° C.; 83%, 0.236 g).
HRMS: Calculated 285.1165; found 285.1172.
To a solution of intermediate 36 (0.14 g, 0.5 mmol) in CH2Cl2 (4 mL) at 0° C. a saturated solution (aq) of NaHCO3 (2 mL) was added. After the addition of methylchloroformate (1.5 eq), the reaction mixture was stirred vigorously at 0° C. for 20 minutes, warmed up to room temperature and allowed to stir further for 0.5 hour. The organic layer was separated, dried over MgSO4 and purified by flash chromatography using EtOAc:heptane (4:6) as an eluant to obtain intermediate 37 as a thick viscous liquid (83%, 0.14 g).
HRMS: Calculated 343.1220; found 343.1218.
To a solution of P(Ph)3 (0.26 g, 1 mmol) in dry THF (4 mL) at −15° C. a solution of DIAD (0.22 g, 1.1 mmol) in THF (1 mL) was added and the resulting complex was stirred for 20 minutes. After the addition of intermediate 37 (0.17 g, 0.5 mmol) dissolved in THF (1 mL) and DPPA (0.14 g, 0.5 mmol) in THF (1 mL), the reaction was warmed to up room temperature and stirred for 18 hours. An excess of P(Ph)3 (5 eq) and water (0.5 mL) was added to the reaction mixture and then heated at 40° C. for 3 hours to reduce the azide to amine functionality. Silica was added to the reaction mixture and the solvent was removed under reduced pressure followed by purification of the product by flash chromatography using CH2Cl2:MeOH (9:1) as an eluant to obtain intermediate 38 as a thick viscous liquid (80%, 0.14 g).
HRMS: Calculated 342.1380; found 342.1376.
To a solution of intermediate 36 (0.5 mmol, 0.14g), Et3N (5 eq) and DMAP (20 mol %) in CH2Cl2 at −20° C., o-nitrobenzenesulphonyl chloride (3 eq) was added. Reaction mixture was warmed up to room temperature and left for overnight stirring. Aqueous NaHCO3 (2 mL) was added to the reaction mixture and the organic layer was separated and dried over MgSO4. Following chromatography (SiO2) using EtOAc:heptane (1:1) as an eluant yielded intermediate 39 as a yellow crystalline solid (mp: 88-90° C., 71%, 0.23 g).
To a solution of acetate diol intermediate 23 (0.12 g, 0.355 mmol) in dry toluene (10 mL) was added n-Bu2SnO (9 mg, 0.036 mmol), Et3N (0.13 mL, 0.888 mmol) and TsCl (0.10 g, 0.533 mmol). Stir at room temperature for 24 hours, add NH4Cl (sat. aq. solution, 10 mL), extract with CH2Cl2 (3×10 mL) and dry with MgSO4. Column purification on silica gel using EtOAc/hexane (30:70) yielded intermediate 23a as a colorless oil (0.15 g, 84%).
Mass spectrum: CI m/z (assignment, relative intensity) 481 (MH+—H2O, 1%), 439 (MH+—AcOH, 4%), 421 (MH+—AcOH—H2O, 1%), 267 (MH+—AcOH-TsOH, 18%), 249 (MH+—AcOH-TsOH—H2O, 100%); EI: m/z (assignment, relative intensity) 480 (M+.—H2O, 1%), 438 (M+.—AcOH, 36%), 266 (M+—AcOH-TSOH, 15%), 248 (M+.—AcOH-TsOH—H2O, 18%); High resolution EI Calculated C25H23FO4S (M+.—AcOH): 438.1301, Found: 438.1300 (51%).
To a solution of tosylate intermediate 23a (1.30 g, 2.61 mmol) in DMF (25 mL) was added NaN3 (0.51 g, 7.83 mmol). The reaction mixture was heated at 100° C. for 1 night. After cooling, add NH4Cl (sat. aq. sol.), extract 3 times with CH2Cl2 and dry with MgSO4. After evaporation the residue was purified on silica gel using EtOAc/heptane (20/80) to give intermediate 40 as an oily product (0.79 g, 82%).
A solution of acetate intermediate 40 (454.9 mg, 1.23 mmol) in MeOH (10 mL) was treated with K2CO3 (340.1 mg, 2.46 mmol) and the mixture was stirred at room temperature for 1 hour. Add NH4Cl (sat. aq. sol.), extract 3 times with CH2Cl2 and dry with MgSO4. The solution was filtered and evaporated and the residue was purified by column chromatography on silica gel using EtOAc/heptane (30/70) to give diol intermediate 41 (370.9 mg, 92%).
To intermediate 41 (670 mg, 2.05 mmol) in CH2Cl2 (25 mL) was added Et3N (2.30 mL, 16.4 mmol), DMAP (0.13 mg, 1.02 mmol) and (CH3SO2)2O (1.07 g, 6.15 mmol) at 0° C. Stir at room temperature for 1 hour, cool to 0° C. again, add AcSH (0.44 ml, 6.15 mmol) and stir at room temperature for 4 hours. Work up by adding NH4Cl (sat. aq. sol.). Extract 3 times with CH2Cl2. Column chromatography on silica gel using CH2Cl2 (100%) afforded intermediate 42 as an oil (0.68 g, 72%).
To the above intermediate 42 (0.15 g, 0.33 mmol) in MeOH (5 mL) was added K2CO3 (92 mg, 0.67 mmol). After stiring at room temperature for 1 night, the mixture was worked up by adding NH4Cl (sat. aq. sol.). Extract 3 times with CH2Cl2 and dry with MgSO4. Column purification on silica gel using CH2Cl2/heptane (40/60) gave intermediate 43 as an oily product (76 mg, 70%).
Mass spectrum: CI m/z (assignment, relative intensity) 326 (MH+, 25%), 298 (MH+—N2, 60%), 283 (MH+—HN3, 100%), 269 (MH+—N2—CH2NH, 12%), 249 (MH+—HN3—H2S, 25%), 235 (MH+—N2—CH2NH—H2S, 21%), 197 (61%).
To a solution of azide intermediate 43 (76.1 mg, 0.23 mmol) in CH2Cl2 (5 mL) was added m-CPBA (m-chloroperbenzoic acid) (173.2 mg, 0.70 mmol). The mixture was stirred at room temperature for 15 min. Add NaHCO3 (sat. aq. solution), extract 3 times with CH2Cl2. Column purification on silica gel using EtOAc/heptane (50/50) gave sulfone intermediate 44 (73.2 mg, 88%) as an oil.
Mass spectrum: —CI m/z (assignment, relative intensity) 358 (MH+, 21%), 340 (MH+—H2O, 9), 330 (MH+N2, 9%), 303 (8%), 265 (24%), 264 (MH+ —N2—H2SO2, 25%), 237 (MH+—N2—H2SO2—HCN, 11%), 211 (15%), 197(66%).
To a solution of intermediate 40 (0.85 g, 2.32 mmol) in THF (10 mL) was added Ph3P (1.22 g, 4.63 mmol) and DIAD (1.92 ml, 4.63 mmol). Then, a solution of p-nitrobenzoic acid (0.77 g, 4.63 mmol) in THF (10 mL) was added dropwise. The mixture was stirred at room temperature for 2 hours. Work up by adding NH4Cl (sat. aq. sol.), extract 3 times with CH2Cl2. Column purification on silica gel using CH2Cl2/heptane (70/30) gave the p-nitrobenzoate (inverted secondary OH group) as an oil (1.19 g, 99%). To a solution of this compound (2.01 g, 4.05 mmol) in MeOH (50 mL) was added K2CO3 (1.12 g, 8.10 mmol). The reaction mixture was stirred at room temperature for 3 hours. Add NH4Cl (sat. aq. sol.), extract 3 times with CH2Cl2. Column purification on silica gel using EtOAc/heptane (30/70) gave an oily intermediate 45 (0.71 g, 98%).
To a solution of the diol intermediate 45 (1.20 g, 3.66 mmol) in CH2Cl2 (30 mL) was added Et3N (4.10 mL, 29.3 mmol), DMAP (0.22 mg, 1.83 mmol) and (CH3SO2)2O (1.92 g, 11.0 mmol) at 0° C. Stir at room temperature for 1 hour. Cool to 0° C. again and add AcSH (0.52 mL, 7.33 mmol) and stir at room temperature for 5 hours. Work up by adding NH4Cl (sat. aq. sol.), extract 3 times with CH2Cl2 and dry with MgSO4. Column purification on silica gel using EtOAc/heptane (30/70) afforded intermediate 46 as an oil (1.32 g, 78%).
To a solution of the above intermediate 46 (1.32 g, 2.86 mmol) in MeOH (30 mL) was added K2CO3 (0.79 g, 5.72 mmol). After stirring at room temperature for 2 hours, NH4Cl (sat. aq. sol.) was added. Extract 3 times with CH2Cl2 and dry with MgSO4. Column purification on silica gel using CH2Cl2/heptane (40/60) gave intermediate 47 as an oily product (0.82 g, 89%).
Mass spectrum: —CI m/z (assignment, relative intensity) 326 (MH+, 25%), 298 (MH+—N2, 60%), 283 (MH+—HN3, 100%), 269 (MH+—N2—CH2NH, 12%), 269 (MH+—HN3—H2S, 25%), 235 (MH+—N2—CH2NH—H2S, 21%), 197 (61%).
To a solution of azide intermediate 47 (136.1 mg, 0.41 mmol) in CH2Cl2 (10 mL) was added m-chloroperbenzoic acid (310.0 mg, 1.26 mmol). The mixture was stirred at room temperature for 30 minutes. Add NaHCO3 (sat. aq. solution), extract 3 times with CH2Cl2. Column purification on silica gel using EtOAc/heptane (50/50) gave sulfone intermediate 48 (146.5 mg, 98%) as an oil.
Mass spectrum: —CI m/z (assignment, relative intensity) 358 (MH+, 21%), 340 (MH+—H2O, 9%), 330 (MH+—N2, 9%), 303 (8%), 265 (24%), 264 (MH+—N2—H2SO2, 25%), 237 (MH+—N2—H2SO2—HCN, 1%), 211 (15%), 197(66%).
To a solution of azide intermediate 43 (0.34 g, 1.05 mmol) in hexafluoroisopropanol (5 mL) was added H2O2 (30%, 0.24 mL, 2.10 mmol). The mixture was stirred at room temperature for 30 minutes. Add Na2CO3 (sat. aq. solution), extract 3 times with CH2Cl2. Column purification on silica gel using Et2O (100%) afforded intermediates 49 (110 mg) and 50 (130 mg) with a total yield of 78%.
Mass spectrum: —CI m/z (assignment, relative intensity) 342 (MH+, 100%), 314 (MH+ —N2, 49%), 299 (MH+—HN3, 47%), 264 (17%), 197 (96%).
To a solution of azide intermediate 47 (0.21 g, 0.64 mmol) in hexafluoroisopropanol (3 mL) was added H2O2 (30%, 0.15 mL, 1.27 mmol). The mixture was stirred at room temperature for 30 minutes. Add Na2CO3 (sat. aq. solution), extract 3 times with CH2Cl2. Column purification on silica gel using Et2O (100%) gave intermediate 51 (120 mg) and 52 (86 mg) with a total yield of 95%.
Mass spectrum: —CI m/z (assignment, relative intensity) 342 (MH+, 100%), 314 (MH+ —N2, 49%), 299 (MH+—HN3, 47%), 264 (17%), 197 (96%).
Dissolve alcohol intermediate 55 (1.72 g, 6.42 mmol) in CH2Cl2 (30 mL). Add Et3N (1.79 mL, 12.8 mmol), DMAP (0.78 g, 6.42 mmol) and AcOH anhydride (1.21 mL, 12.8 mmol). Stir at room temperature for 1 hour and add sat. aq. NH4Cl (15 mL). Extract 3 times with CH2Cl2 (3×20 mL) and dry with MgSO4. Column purification on silica gel using CH2Cl2/hexane (60:40) yielded intermediate 56 as an oil (1.77 g, 89%).
Mass spectrum: —CI m/z (assignment, relative intensity) 311 (MH+, 5%), 251 (MH+—AcOH, 100%); EI: m/z (assignment, relative intensity) 250 (M+.—AcOH, 16%), 209 (M−. —AcOH—CH2CH2—CH2, 100%); High resolution EI Calculated C18H15F (M+. —AcOH): 250.1158, Found: 250.1162 (26%).
A mixture of the diol intermediate 5 (6.022 g, 18.40 mmol), Et3N (5.586 g, 55.2 mmol), 4-dimethylaminopyridine (138 mg, 1.13 mmol), trityl bromide (9.444 g, 27.6 mmol) in CH2Cl2 (180 mL) was stirred at room temperature under nitrogen atmosphere for 2 hours, then quenched with saturated aqueous NH4Cl (50 mL). The organic phase was separated, aqueous layer extracted with CH2Cl2 (2×50 mL), combined organics washed with water (3×40 mL), brine (40 mL), dried (MgSO4) and evaporated in vacuo. Purification by flash chromatography (Kieselgel 60, 230-400 mesh, EtOAc-heptane, 5/95 to 10/90) gave intermediate 57 (8.595 g, 15.09 mmol, 82%) as a brown semisolid.
A mixture of the alcohol intermediate 57 (8.500 g, 14.92 mmol), Et3N (4.529 g, 44.76 mmol) and DMAP (84 mg, 0.689 mmol) in CH2Cl2 (200 mL) was cooled down to −78° C. under nitrogen atmosphere. MsCl (2.264 g, 22.38 mmol) was added in one portion, resulting solution slowly warmed up to room temperature (ca. 40 min) and quenched with saturated aqueous NH4Cl (50 mL). The organic phase was separated, aqueous layer extracted with CH2Cl2 (3×45 mL), combined organics washed with water (3×45 mL) and brine (40 mL), dried (MgSO4), and evaporated in vacuo. Due to the instability of intermediate 58 it was used immediately without further purification.
Crude intermediate 58 (unknown amount, assumed 14.92 mmol), was dissolved in MeOH (200 mL), dry Amberlyst-15 (15 g) added and the mixture was stirred at 45° C. for 4 hours; progress of reaction followed by TLC (Kieselgel on glass; EtOAc-heptane 30/70). The resin was filtered off and washed with MeOH (2×40 mL), methanolic solution concetrated in vacuo to 100 mL, and intermediate 59 used immediately for the next step.
Methanolic solution of intermediate 59, obtained as above, was treated with anhydrous K2CO3 (4.146 g, 30 mmol) and stirred at room temperature for 3 hours. After treatment with water (100 mL), MeOH was removed in vacuo, product extracted with Et2O (3×75 mL). The combined organics were washed with water (3×75 mL) and brine (40 mL), dried (MgSO4) and evaporated in vacuo. Chromatographic purification (Kielselgel 60, 70-230 mesh, EtOAc-heptane 10/90) gave intermediate 60 (3.185 g, 10.29 mmol, 69% from intermediate 57) as a colorless oil.
HRMS: Calcd. for C18H16FN3O: 309.1277; Found: 309.1279.
Epoxide intermediate 60 (3.108 g, 10.04 mmol) was dissolved in MeOH (50 mL), Et3N (1.012 g, 10 mmol) and 10% Pd—C (150 mg) added, and resulting mixture hydrogenated under atmospheric pressure for 5 hours. Catalyst was removed by filtration through short pad of Kieselguhr, MeOH and Et3N removed in vacuo, and residue purified by column chromatography (Kieselgel 60, 70-230 mesh, EtOH—CH2Cl2 5/95) to yield intermediate 61 (2.333 g, 8.23 mmol, 82%) as a yellowish oil, slowly solidifying on standing.
HRMS: Calcd. for C18H18FNO: 283.13 72; Found: 283.1380.
Pyrrolidine intermediate 61 (567 mg, 2.00 mmol) was dissolved at 0° C. in a mixture of CH2Cl2 (20 mL) and saturated aqueous NaHCO3 (20 mL), then methyl chloroformate (0.23 mL, 281 mg, 2.98 mmol) was added, ice bath removed, and resulting mixture stirred for 5 hours. The organic layer was separated, aqueous phase extracted with CH2Cl2 (40 mL) then the combined organics washed with water (2×40 mL), brine (20 mL), dried (MgSO4) and evaporated. The residue was purified by column chromatography on silica (CH2Cl2-EtOH, 95/5) to give carbamate intermediate 62 (669 mg, 1.96 mmol, 98%) as tan oil, solidifying on standing.
HRMS: Calcd. for C20H20FNO3: 341.1427; Found: 341.1435.
A mixture of intermediate 61 (283 mg, 1 mmol), ethyl formate (741 mg, 10 mmol) and acetonitrile (10 mL) was refluxed for 18 hours, then evaporated in vacuo. The residue was purified by chromatography (Kieselgel 60, 70-230 mesh, EtOH—CH2Cl2 5/95) to yield 62a (283 mg, 0.91 mmol, 91%) as a tan solid.
Product is mixture of 2 rotamers (3:2 ratio).
CI-MS (CH4) 312 (MH+, 100%); 292 (MH+—HF, 13%).
Pyridinium chlorochromate (104 mg, 0.48 mmol) was added to the solution of alcohol intermediate 62a (100 mg, 0.32 mmol) in CH2Cl2 (10 mL), and the resulting slurry was stirred under nitrogen atmosphere for 3 hours. After addition of Et2O (20 mL), the mixture was filtered through silica, the tar residue in flask washed with Et2O (40 mL), filtered again, evaporated to dryness in vacuo. Crude aldehyde intermediate 63 (77 mg, 0.25 mmol, 78%) was obtained as reddish oil, containing trace of chromium. Product was used immediately without purification.
CI-MS (CH4) 310 (100%, MH+), 290 (11%, MH+—HF); 282 (7%, MH+—CO).
Poly(triphenylphosphine) (0.33 g, ca. 1 mmol of Ph3P) was swollen under argon atmosphere in dry CH2Cl2 (10 mL), then diisopropyl azodicarboxylate (222 mg, 1.1 mmol) in THF (3 mL) was added through septum at 0° C. The suspension was stirred for 30 minutes at 0° C., followed by addition of intermediate 61 (104 mg, 0.366 mmol) in THF (4 mL). The cooling bath was removed and reaction mixture was stirred at room temperature for 12 hours, then water (0.1 mL) was added, resin filtered off and washed with THF (15 mL), combined organics evaporated and purified by column chromatography (Kieselgel 60, 230-400 mesh, CH2Cl2-EtOH 100/0 to 96/4) to give intermediate 64 (63 mg, 0.238 mmol, 65%) as yellowish oil.
HRMS Calcd. for C18H16FN: 265.1267; Found: 265.1270.
A solution of pyrrolidine intermediate 61 (567 mg, 2.00 mmol), Et3N (1.012 mmol, 10.00 mmol), dimethylaminipyridine (40 mg, 0.33 mmol) in CH2Cl2 (30 mL) was treated with 2-nitrobenzenesulfonyl chloride (1.330 g, 6.00 mmol), and resulting mixture was stirred at room temperature for 3 hours, then quenched with saturated aqueous NH4Cl (30 mL). After extraction with CH2Cl2 (3×30 mL) the combined organics were washed with 1N HCl (15 mL), saturated aqueous K2CO3 (40 mL), water (3×40 mL), brine, dried (MgSO4), evaporated and purified by column chromatography on silica (heptane-EtOAc, 95/5→85/15) to give intermediate 65 (1.203 g, 1.84 mmol, 92%) as yellow crystals, rapidly decomposing on standing.
1H NMR (300 MHz, CDCl3) δ 8.26-7.50 (m, 8H, Ar—H, 2-nosyl moieties); 7.20-7.03 (m, 6H, Ar—H, dibenzosuberone part); 6.81 (td, J=8.3, 2.7 Hz, 1H, Ar—H); 5.40 (d, J=11.0 Hz, 1H, CH-12b); 4.69 (d, J=16.7Hz, 1H, CH2-8); 4.60 (m, 2H, CH2ONs); 4.40 (m, 1H, CH-2); 3.73 d, J=16.7Hz, 1H, CH2′-8); 3.55-3.40 (m, 1H, CH-3a); 2.80 (dd, J=13.0, 6.2 Hz, CH2-3); 2.33-2.18 (m, 1H, CH2′-3).
A mixture of bisnosyl intermediate 65 (0.35 g, 0.54 mmol) and appropriate amine (3 mmol) in dioxane (10 mL) was refluxed for 4 hours, cooled down to ambient temperature, diluted with water (100 mL), precipitated product filtered off, washed with water (100 mL), dissolved in EtOAc, solution washed with brine, dried (K2CO3), evaporated, and used for next step without purification.
To a solution of (S)-1,2,4-butanetriol intermediate 67a (6.76 g, 63.68 mmol) in freshly distilled pentan-3-one (320 mL) was added p-toluenesulfonic acid (p-TSA) (6.06 g, 31.84 mmol). The reaction mixture was stirred at 53° C. for 16 hours, then Et3N (10 mL) was added and the reaction mixture stirred at ambient temperature for 10 minutes. The reaction mixture was concentrated under reduced pressure. Gradient flash chromatography (CH2Cl2/MeOH, 100:0 to 97:3 to 95:5) afforded the protected alcohol intermediate 67b (9.84 g, 89%) as a colorless oil.
To a solution of 2(S)-2-(2,2-diethyl-[1,3]dioxolan-4-yl)-ethanol intermediate 67b (4.00 g, 22.96 mmol) and 4A molecular sieves (11.50 g) in CH2Cl2 (200 mL) stirred at 0° C. for 5 minutes was added pyridinium chlorochromate (PCC) (9.90 g, 45.92 mmol). The reaction mixture was allowed to warm to ambient temperature and stirred for 1 hour. The crude reaction mixture was filtered through a plug of silica, washed with Et2O (50 mL) and concentrated under reduced pressure to afford the aldehyde intermediate 67c (3.56 g, 90%) as a colorless oil.
MgBr2 (0.733 g, 3.98 mmol) was added to 8-fluoro-5,11-dihydro-10H-dibenzo[a,d]-cyclohepten-10-one (0.75 g, 3.32 mmol) in toluene (15 mL) and the reaction mixture stirred at room temperature for 30 minutes. Aldehyde intermediate 67c (2.05 g, 11.92 mmol) in THF (10 mL) was added and in one time t-BuOK (0.074 g, 0.66 mmol). The reaction mixture was stirred for 22 hours at ambient temperature, then saturated aqueous NH4Cl (15 mL) was added. The product was extracted three times with Et2O (3×30 mL), combined organics washed with water (2×35 mL), brine (25 mL) dried over MgSO4. After evaporation of the toluene, the residue was purified on silica gel column using Et2O/heptane (10/90) to obtain intermediate 67d (1.079 g, 86%) as a yellowish oil.
HRMS (EI) Calcd. for C24H25FO3: 380.1800; Found: 380.1785.
10% Pd—C (200 mg) and Et3N (0.135 ml, 0.97 mmol) were added to intermediate 67d (0.246 g, 0.647 mmol) in i-PrOH (25 mL) and toluene (15 mL) and subjected to hydrogenation overnight at room temperature. The reaction mixture was dissolved in CH2Cl2, filtered through celite and solvent evaporated. The residue was purified on a silica gel column using Et2O/heptane (30/70) to give intermediate 67e (151 mg, 61%) as a yellowish oil.
HRMS C24H27FO3: 382.1944; Found: 382.1951.
Sodium borohydride (1.78 g, 46.84 mmol) was added to intermediate 35 (2.0 g, 5.88 mmol) dissolved in i-PrOH (80 mL)/pH 7 phosphate buffer (30 ml) at 0° C. After 1 hour of reaction at room temperature, NH4Cl (sat. aq. solution) was added and the mixture was extracted three times with CH2Cl2. The organic phase was dried over MgSO4 and the solvent evaporated. The product was purified on silica gel using Et2O/heptane (30/70) which gave intermediate 67f (1.96 g, 97%) as colorless oil.
HRMS Calcd. for C21H23FO3: 342.1631; Found: 342.1627.
Intermediate 67g was obtained in the same way as described for intermediate 30.
Intermediate 67h was obtained in the same way as described for intermediate 31.
A mixture of (2S)-4-[(10R,11S)-11-azido-2-fluoro-10,11-dihydro-5H-dibenzo[a,d]-cyclohepten-10-yl]-1,2-butanediol intermediate 67h (225 mg, 0.66 mmol), Et3N (506 mg, 5.0 mmol), dimetylaminopyridine (12 mg, 0.1 mmol) and TsCl (503 mg, 2.64 mmol) in CH2Cl2 (25 mL) was stirred at room temperature under nitrogen atmosphere for 15 hours. After quenching with saturated aqueous ammonium chloride (15 mL), the organic phase was separated, and the aqueous layer extracted with CH2Cl2 (3×20 mL). The combined organics were washed with water (3×30 mL), brine (25 mL), dried over magnesium sulfate and evaporated in vacuo. The residue was purified by column chromatography (Kieselgel 60, 70-230 mesh, heptane-ethyl acetate 90/10) to afford intermediate 67i (352 mg, 0.54 mmol, 82%) as colorless semisolid.
Bistosylate intermediate 67i (340 mg, 0.52 mmol) was dissolved in MeOH (15 mL), Et3N (1.012 g, 10 mmol) and 10% Pd—C (150 mg) added, and resulting mixture hydrogenated under atmospheric pressure for 5 hours. Catalyst was removed by filtration through short pad of Kieselguhr, anhydrous K2CO3 (138 mg, 1 mmol) added and resulting slurry stirred at room temperature for 5 hours. After filtration of solids, MeOH and Et3N were removed in vacuo, and residue purified by flash chromatography (Kieselgel 60, 230-400 mesh, EtOH—CH2Cl2 5/95 to 12/88) to yield intermediate 67k (153 mg, 0.338 mmol, 65%) as yellowish oil.
CI-MS (CH4) 452 (MH+, 1%); 280 (MH+-TsOH, 100%).
A solution of triphenylphosphine (910 mg, 3.5 mmol) in dry THF (25 mL) was placed in a two-necked 100 mL flask, equipped with septum, argon inlet and magnetic stirrer. After cooling down to −15° C. neat diisopropyl azodicarboxylate (708 mg, 3.5 mmol) was added through a septum with intensive stirring. Resulting yellow suspension was stirred at above temperature for 30 minutes, then a solution of 4-nitrobenzoic acid (585 mg, 3.50 mmol) and alcohol intermediate 67f (673 mg, 1.75 mmol) in THF (25 mL) was added within 10 minutes. The resulting yellow suspension was allowed to warm up to room temperature and stirred then for 12 hours. Water (0.3 mL) was added, followed by silica gel (Kieselgel 60, 70-230 mesh, 4 g), THF removed in vacuo, and silica powder submitted to the flash chromatography (Kieselgel 60, 230-400 mesh, EtOAc-heptane, 5/95 to 15/85) to give nitrobenzoate intermediate 610 (795 mg, 1.49 mmol, 85%) as an orange semisolid.
Hydrolysis of dioxolane intermediate 610 (795 mg, 1.49 mmol) was carried out in the same was as described in Example A28 to give diol intermediate 68b (695 mg, 1.49 mmol, 100%) as orange semisolid. Product was used without purification.
A mixture of the diol intermediate 68b (695 mg, 1.49 mmol), Et3N (607 mg, 6 mmol), dibutyl(oxo)stannane (141 mg, 0.566 mmol), and TsCl (431 mg, 2.26 mmol) in CH2Cl2 (20 mL) was stirred at room temperature under N2 for 12 hours. After quenching with saturated aqueous NH4Cl (15 mL) the organic phase was separated and the aqueous solution extracted with CH2Cl2 (3×30 mL). The combined organics were washed with water (3×20 mL), filtered through 5 cm layer of MgSO4, and evaporated in vacuo to furnish crude intermediate 68c (601 mg, 0.97 mmol, 65%) as a yellowish semisolid mass, which was converted without further purification.
A mixture of tosylate intermediate 68c (601 mg, 0.97 mmol), sodium methoxide (162 mg, 3.0 mmol) and MeOH (10 mL) was stirred at room temperature for 3 hours. After treatment with water (100 mL) product was extracted with diethyl ether (3×30 mL). The combined organics were washed with water (3×40 mL) and brine (40 mL), dried (MgSO4) and evaporated in vacuo. Chromatographic purification (Kielselgel 60, 70-230 mesh, EtOAc-heptane 10/90 to 25/75) gave intermediate 68d (211 mg, 0.71 mmol, 73%) as a colorless oil.
HRMS Calcd. for C19H10FO2: 298.1369; Found 298.1350.
A mixture of the alcohol intermediate 68d (211 mg, 0.708 mmol), Et3N (209 μL, 287 mg, 2.83 mmol), DMAP (86.5 mg, 0.708 mmol), and TsCl (270 mg, 1.42 mmol) in CH2Cl2 (10 mL) was stirred at room temperature under N2 for 16 hours. After quenching with saturated aqueous NH4Cl (10 mL) the organic phase was separated and the aqueous solution extracted with CH2Cl2 (3×15 mL). The combined organics were washed with water (3×15 mL), dried (MgSO4), and evaporated in vacuo to give crude intermediate 68e (282 mg, 0.62 mmol, 88%) as a yellowish oil, which was used without further purification.
A solution of triphenylphosphine (1049 mg, 4.0 mmol) in dry THF (20 mL) was placed in a two-necked 100 mL flask, equipped with septum, argon inlet and magnetic stirrer. After cooling down to −15° C. neat diisopropyl azodicarboxylate (809 mg, 4.0 mmol) was added through a septum with intensive stirring. Resulting yellow suspension was stirred at above temperature for 30 minutes, then a solution of 4-nitrobenzoic acid (4.0 mmol) and 11-(2,2-dimethyl-[1,3]dioxolan-4-ylmethyl)-8-fluoro-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-10-ol (intermediate 3) (685. mg, 2.0 mmol) in THF (25 mL) was added within 10 minutes. The resulting yellow suspension was allowed to warm up to room temperature and stirred then for 12 hours. Water (0.3 mL) was added, followed by silica gel (Kieselgel 60, 70-230 mesh, 4 g), THF removed in vacuo, and silica powder submitted to the flash chromatography (Kieselgel 60, 230-400 mesh, EtOAc-heptane, 5/95 to 10/90) to give nitrobenzoate intermediate 69a (875 mg, 1.78 mmol, 89%) as an orange semisolid.
Intermediate 69b has been obtained from acetal intermediate 69a (860 mg, 1.75 mmol) in the same way as described for intermediate 5. Column chromatography (Kieselgel 60, 70-230 mesh, EtOAc-heptane, 35/65 to 50/50) afforded diol intermediate 69b (774 mg, 1.715 mmol, 98%) as ayellow semi-solid.
Diol intermediate 69b (774 mg, 1.715 mmol) was dissolved at 0° C. in a mixture of THF (25 mL) and pH 7 phosphate buffer (5 mL), then sodium periodate (642 mg, 3 mmol) was added in one portion at 0° C., cooling bath removed, and resulting mixture was stirred at room temperature for 4 hours. Water (50 mL) was added, product extracted with diethyl ether (3×30 mL). The combined organics were washed with saturated aqueous sodium metabisulfite (50 mL), water (2×50 mL), brine (30 mL), dried over magnesium sulfate and evaporated in vacuo to give aldehyde intermediate 69c (680 mg, 1.66 mmol, 97%) as a yellow foam. Product was used immediately without purification.
A mixture of aldehyde intermediate 69c (680 mg, 1.66 mmol), AcOH (240 mg, 4.0 mmol), bis(dimethylamino)methane (719 mg, 7.0 mmol) and THF (30 mL) was stirred at room temperature for 3 hours. Water (50 mL) was added, product extracted with diethyl ether (3×30 mL). The combined organics ware washed with saturated aqueous sodium bicarbonate (25 mL), water (2×50 mL), brine (30 mL), dried over magnesium sulfate and evaporated in vacuo to give unsaturated aldehyde intermediate 69d (680 mg, 1.58 mmol, 95%) as a yellow oil.
Sodium borohydride (190 mg, 5.00 mmol) was added at room temperature within 10 minutes to the solution of aldehyde intermediate 69d (680 mg, 1.58 mmol) in MeOH (30 mL). The reaction mixture was stirred at room temperature for 4 hours, quenched with saturated aqueous ammonium chloride (20 mL) and extracted with Et2O (3×30 mL). The combined organics ware washed with water (2×50 mL), brine (30 mL), dried over magnesium sulfate and evaporated in vacuo to give alcohol intermediate 69e (582 mg, 1.34 mmol, 85%) as an orange oil.
A mixture of nitrobenzoate intermediate 69e (582 mg, 1.34 mmol), sodium methoxide (162 mg, 3.0 mmol) and MeOH was stirred at room temperature for 4 hours. Water (70 mL) was added, product extracted with EtOAc (3×30 mL). The combined organics were washed with water (2×50 mL), brine (30 mL), dried over magnesium sulfate and evaporated in vacuo. The residue was purified by flash chromatography (Kieselgel 60, 230-400 mesh, EtOAc-heptane, 35/65 to 60/40) to give diol intermediate 69f (286 mg, 1.005 mmol, 75%) as colorless oil.
Tributylphosphine (405 mg, 2.0 mmol) was dissolved in toluene (25 mL) under argon atmosphere. Diisopropyl azodicarboxylate (405 mg, 2.0 mmol) in toluene (3 mL) was added dropwise, followed by solution of diol intermediate 69f (270 mg, 0.95 mmol). The resulting mixture was stirred at room temperature for 3 hours, then reaction was quenched water (1 mL). Silica gel (Kieselgel 60, 70-230 mesh, 1.3 g) was added, toluene removed in vacuo, and silica powder submitted to the flash chromatography (Kieselgel 60, 230-400 mesh, EtOAc-heptane, 5/95 to 12/88) to give THF derivative intermediate 69g (205 mg, 0.77 mmol, 81%) as colorless foam.
Boron trifloride etherate (0.43 mL, 3.54 mmol) in THF (1 mL) was added at room temperature under argon atmosphere to the solution of THF intermediate 69g (188 mg, 0.66 mmol), sodium borohydride (496 mg, 2.64 mmol) in dry THF (2 mL). The resulting solution was stirred under argon for 24 hours, excess of borohydrided decomposed carefully with water (3.8 mL), MeOH(1.5 mL) added, followed by 3M NaOH (3.8 mL) and 30% hydrogen peroxide (0.55 mL). Reaction mixture was allowed to stir for 4 hours at room temperature, then the product was extracted with Et2O (3×30 mL). The combined organics ware washed with water (2×50 mL), brine (30 mL), dried over magnesium sulfate and evaporated in vacuo. The residue was purified by flash chromatography (Kieselgel 60, 230-400 mesh, EtOAc-heptane, 5/95 to 20/80) to give THF derivative intermediate 69h (139 mg, 0.49 mmol, 74%) as colorless oil.
Poly(triphenylphosphine) (0.33 g, ca. 1 mmol of Ph3P) was swollen at room temperature under argon atmosphere in dry THF (10 mL), then diisopropyl azodicarboxylate (222 mg, 1.1 mmol) in THF (3 mL) was added through septum at −15° C. The suspension was stirred for 30 minutes at −15° C., then alcohol intermediate 69h (139 mg, 0.49 mmol) in dry THF (2.5 mL) was added in one portion, followed by dropwise addition of diphenylphosporyl azide (160 mg, 0.58 mmol) in THF (3 mL). Resulting suspension was stirred under argon for 12 hours. After quenching with water (0.3 mL), resin was filtered off and solvent removed in vacuo. The residue was purified by flash chromatography (Kieselgel 60, 230-400 mesh, EtOAc-heptane, 15/85) to give azide intermediate 69i (136 mg, 0.44 mmol, 90%) as colorless foam.
Triol intermediate 70a was obtained from intermediate 3 (514 mg, 1.50 mmol) in the same way as described in Example A5. Crude intermediate 70a (449 mg, 1.485 mmol, 99%) was obtained as colorless oil and used without purification.
Intermediate 70b was obtained from triol intermediate 70a (449 mg, 1.485 mmol) in the same way as described for compound 44. Flash chromatography (Kieselgel 60, 230-400 mesh, EtOAc-heptane, 10/90 to 33/67) afforded intermediate 70c (357 mg, 1.32 mmol, 89%) as tan solid.
Reaction of intermediate 70c (335 mg, 1.24 mmol) was carried out in the same way as described for compound 45. Complex, unseparable mixture of products has been formed and used for the next step without purification.
Reaction of the mixture containing intermediate 70c was was carried out in the same way as described for compound 46. Purification by RP-HPLC (Waters Xterra® C18, 19×50 mm, MeOH-water 50/50, then pure MeOH, 4 mL/min) afforded intermediate 70d (135 mg, 0.41 mmol, 33% from intermediate 70b) as yellow oil.
Reaction of diol intermediate 5 (0.99 g, 3.02 mmol) was carried out in the same way as described for compound 44. Purification by column chromatography (Kieselgel 60, 230-400 mesh, diethyl ether-heptane, 50/50) gave aldehyde intermediate 71a (778 mg, 2.63 mmol, 87%) as colorless oil.
Reaction of intermediate 71a (618 mg, 2.09 mmol) was carried out in the same way as described for compound 45. Crude aldehyde intermediate 71b (605 mg, 1.97 mmol, 94%) was obtained as colorless oil and was used without further purification.
Poly(triphenylphosphine) (1.40 g, ca. 4.2 mmol of Ph3P) was swollen at room temperature under argon atmosphere in THF (30 mL), then aldehyde intermediate 71b (405 mg, 1.32 mmol) in THF (10 mL) and water (0.19 g) were added. The resulting mixture was stirred under argon at 50° C. for 1 hour. After this time resin was filtered off, THF remove in vacuo. The residue was dissolved in MeOH (10 mL), AcOH (1 mL) and sodium cyanoborohydride (200 mg, 3.2 mmol) added and resulting mixture stirred at room temperature for 2 hours, then quenched with concentrated HCl (1 mL), treated with saturated aqueous NaHCO3 (15 mL) and basified with 1N sodium hydroxide (3 mL). Product was extracted with CH2Cl2 (3×50 mL), combined organics washed with water (2×30 mL), brine (30 mL), dried (MgSO4) and evaporated in vacuo to afford pyrrolidine intermediate 71c (258 mg, 0.97 mmol, 74%) as yellow foam. Intermediate 71c was used without further purification.
Reaction of intermediate 71c (258 mg, 0.97 mmol) was carried out in the same way as described for compound 9. Flash chromatography (Kieselgel 60, 230-400 mesh, heptane-EtOAc 50/50 to 0/100) afforded intermediate 71d (282 mg, 0.87 mmol, 90%) as yellow oil.
Reaction of 71d (255 mg, 0.79 mmol) was carried out obtained in the same way as described for compound 49. Flash chromatography (Kieselgel 60, 230-400 mesh, EtOH—CH2Cl2 1/99 to 3/97) afforded 71e (215 mg, 0.63 mmol, 80%) as colorless oil.
Reaction of intermediate 71f (215 mg, 0.63 mmol) was carried out obtained in the same way as described for compound 50. Flash chromatography (Kieselgel 60, 230-400 mesh, ethyl acetate) afforded intermediate 71f (194 mg, 0.53 mmol, 84%) as colorless oil.
B. Preparation of the Final Compounds.
The compounds prepared hereinunder all are mixtures of isomeric forms, unless otherwise specified.
To a solution of diamine intermediate 9 (130 mg, 0.3 mmol) in MeOH (5 mL) was added Et3N (126.5 μL, 0.91 mmol) and the mixture was hydrogenated at 1 atmospheric pressure with 10% palladium-on-charcoal under vigorous stirring at room temperature. After 1 hour, formaldehyde (112.8 μL, 1.5 mmol) was added and the mixture was hydrogenated for an additional hour. The suspension was then filtered through a pad of celite and the solids were washed 4 times with CH2Cl2. After evaporation, the crude product was purified by column chromatography on silica gel using CHCl3/MeOH (95/5). This yielded final compound 1 as an oil (50.5 mg, 54%).
Mass spectrum: —CI m/z (assignment, relative intensity) 309 (MH+, 100%), 289 (MH+—HF, 26%); EI: m/z (assignment, relative intensity) 308 (M+, 68%), 279 (M−. —CH2NH, 4%), 265 (M+.CH3NCH2, 100%), 197 (23%); High resolution EI Calculated C20H21FN2 (M+.): 308.1689, Found: 308.1684 (35%).
Dissolve the above compound 1 (0.114 g, 0.37 mmol) in MeOH (10 mL) and add TFA (0.071 mL, 0.93 mmol), NaCNBH3 (0.058 g, 0.93 mmol) and stir at room temperature for 1 hour. Add 10 mL K2CO3 (sat. aq. solution), extract with CH2Cl2 (3×10 mL) and dry with MgSO4. Column purification on silica gel using CH2Cl2/MeOH (10%) gave final compound 2 as an oil (0.067 g, 59%).
Mass spectrum: —CI m/z (assignment, relative intensity) 311 (MH+, 100%), 291 (MH+—HF, 25%), 282 (MH+—CH2NH), 266 (MH+—HN(CH3)2, 13%), 252 (8%); EI: m/z (assignment, relative intensity) 310 (M+., 26%), 266 (M+ —(CH3)2N, 76%), 252 (M+.—(CH3)2NCH2, 70%), 235 (100%), 209 (61%); High resolution EI Calculated C20H23FN2 (M+.): 310.1845, Found: 310.1820 (5%).
To solution of the diamine intermediate 9 (238.6 mg, 0.85 mmol) in DMF (3 mL) was added carbon disulfide (0.076 mL, 1.28 mmol). Stir at 60° C. for 20 minutes. After evaporation of the solvent, the residue was purified by column chromatography on silica gel using EtOAc/heptane (50/50) to give compound 3 as a semisolid final compound (124.6 mg, 45%).
Mass spectrum: —CI m/z (assignment, relative intensity) 325 (MH+, 100%), 252 (1%), 224 (2%).
A solution of the above carbamate intermediate 16 (86.2 mg, 0.19 mmol) in MeOH (3 mL) was hydrogenated at 1 atmospheric pressure with 10% palladium-on-charcoal under vigorous stirring at room temperature. After reaction for 1 hour, formaldehyde (70.7 μL, 0.94 mmol) was added and the mixture was hydrogenated for an additional hour. The suspension was filtered through a pad of celite and the solids were washed 4 times with CH2Cl2. After evaporation of the solvent, the crude product was purified by column chromatograhy on silica gel using CHCl3 to yield final compound 4 (18.6 mg, 29%).
Mass spectrum: —CI m/z (assignment, relative intensity) 337 (MH+, 100%), 317 (MH+—HF, 18%), 309 (MH+—CO, 9%), 161 (9%), 133 (75%), 93 (72%).
To a solution of diamine intermediate 13 (28.7 mg, 0.05 mmol) in MeOH (2 mL) was added Et3N (21.4 μL, 0.15 mmol) and formaldehyde (18.8 μL, 0.25 mmol) and the mixture was treated with hydrogen under 1 atmospheric pressure and 10% palladium-on-charcoal under vigorous stirring at room temperature. After reaction for 1 hour, the suspension was filtered through a pad of celite and the solids were washed 4 times with CH2Cl2. After evaporation, the crude product was purified by column chromatograhy on silica gel using CHCl3/MeOH (90/10). This afforded final compound 5 as an oil (16.7 mg, 98%).
Mass spectrum: —CI m/z (assignment, relative intensity) 325 (MH+, 100%), 323 (25%), 305 (MH+—HF, 19%), 280 (MH+—HN(CH3)2, 12%), 266 (MH+—CH3N(CH3)2, 36%).
To a solution of diamine intermediate 13 (40.4 mg, 0.14 mmol) in CH3CN (2 mL) was added Et3N (50 μL, 0.36 mmol) and the mixture was heated under argon at 70° C. After 1 hour, a solution of diphenyl carbonate (3 6.6 mg, 0.17 mmol) in CH3CN was added dropwise and the mixture was stirred at 70° C. for 2 days. After evaporation, the crude product was purified by column chromatography on silica gel using EtOAc/heptane (20/80) to yield urea final compound 6 as an oil (23 mg, 52%).
Mass spectrum: —CI m/z (assignment, relative intensity) 309 (MH+, 100%), 308 (12%), 289 (MH+—HF, 20%), 279 (3%), 113 (8%).
To a solution of urea compound 6 (29 mg, 0.10 mmol) in THF (3 mL) was added NaH (15.9 mg, 0.31 mmol) and the mixture was stirred at room temperature for 20 minutes. Then Me2SO4 (25.4 mg, 0.26 mmol) was added and the mixture was stirred for an additional 30 minutes. Add 10 mL of NH4Cl (sat. aq. solution), extract with CH2Cl2 (3×10 mL) and dry with MgSO4. Column purification on silica gel using EtOAc/heptane (40/60) gave the methylated urea final compound 7 as an oil (19 mg, 63%).
Mass spectrum: —CI m/z (assignment, relative intensity) 323 (MH+, 100%), 303 (MH+—HF, 26%), 209 (2%), 127 (3%).
To a solution of the diamine intermediate 13 (54 mg, 0.19 mmol) in DMF (3 mL) was added carbon disulfide (17.3 μL, 0.29 mmol). After stirring at 60° C. for 20 minutes, followed by evaporation of the solvent, column purification on silica gel (eluent: EtOAc/heptane (50/50)) gave a crystalline final compound 8 (27.3 mg, 44%); mp: 150-151° C.
Mass spectrum: —CI m/z (assignment, relative intensity) 325 (MH+, 100%), 252 (1%), 224 (2%).
To a solution of final compound 8 (140.6 mg, 0.43 mmol) in MeOH (10 mL) was added methyl iodide (53.5 μL, 0.86 mmol) and Et3N (129 μl, 0.86 mmol). After stirring at 80° C. for 2 days, solvent and reagents were evaporated. Column purification on silica gel (eluent: EtOAc/heptane (40/60)) gave the S-methylated final compound 9 as an oil (72.7 mg, 49%).
Mass spectrum: —CI m/z (assignment, relative intensity) 339 (MH+, 100%), 319 (MH+—HF, 4%), 268 (8%), 266 (3%).
To a solution of intermediate 19 (265.3 mg, 0.44 mmol) in MeOH (20 mL) was added MeSO3H (2 mL) and the mixture was stirred at 60° C. for 30 minutes. After evaporation of the solvent, NaHCO3 (sat. aq. solution) (15 mL) was added and the mixture was extracted with CH2Cl2 (3×10 mL). The combined organic layers were dried with MgSO4. Column purification on silica gel using CH2Cl2/MeOH (5%) gave the amino compound (125 mg, 79%). The latter was then dissolved in MeOH (30 mL). Following addition of formaldehyde (80 μL, 1.06 mmol) the mixture was hydrogenated (1 atm.) with 10% palladium-on-charcoal under vigorous stirring at room temperature for 6 hours. The suspension was then filtered through a pad of celite and the solids were washed 4 times with CH2Cl2. After evaporation of the solvent, the crude product was purified by column chromatograhy on silica gel using CHCl3/MeOH (95/5). Final compound 10 (90.1 mg, 67%) was obtained as an oil (mixture of conformers).
Mass spectrum: -APCI m/z (assignment, relative intensity) 383 (MH+, 100%), 369 (4%), 367 (4%), 363 (MH+—HF, 5%), 354 (2%), 351 (2%).
Intermediate 21 (53.4 mg, 0.15 mmol) was dissolved in a sat. solution of HCl in MeOH (10 mL) and the mixture was stirred at 60° C. overnight. After removal of solvent, 10 mL of K2CO3 (sat. aq. solution) was added and the mixture extracted with CH2Cl2 (3×10 mL). Column purification on silica gel using CHCl3/MeOH (97/3) gave the amino ester compound 11 (20 mg, 37%) as an oil.
Mass spectrum: —CI m/z (assignment, relative intensity) 355 (MH+, 100%), 335 (MH+—HF, 14%), 295 (MH+—CH3OH—CO, 4%), 252 (MH+—CH3OH—CH2CO—NHCH2, 8%), 169 (5%), 141 (46%); EI m/z (assignment, relative intensity) 354 (M, 3%), 295 (M−—CH3OCO, 4%), 252 (M+—CH3OCOCH2NHCH2, 100%), 235 (M+—CH3OCOCH2NHCH2—NH3, 68%), 223 (8%), 209 (22%); High resolution EI Calculated C21H23N2O2F (M+): 354.1744,Found: 354.1751 (9%).
Intermediate 21 (250 mg, 0.71 mmol) was dissolved in 10 mL of HCl in MeOH (sat solution) and the mixture was stirred at room temperature overnight. The reaction was quenched by addition of 10 mL of K2CO3 (sat. aq. solution). The mixture was then extracted 3 times with 10 mL CH2Cl2. The combined organic layers were dried over MgSO4 and evaporated. Column purification on silica gel using CHCl3/MeOH (95/5) gave final compound 12 (67.6 mg, 29%) as an oil.
Mass spectrum: —CI m/z (assignment, relative intensity) 323 (MH+, 100%), 303 (MH+—HF, 20%), 295 (MH+—CO, 2%), 252 (MH+—COCH2—NHCH2, 1%), 188 (2%), 160 (5%); EI m/z (assignment, relative intensity) 322 (M+, 100%), 252 (M+—COCH2N═CH2, 40%), 235 (68%), 223 (M−—COCH2N═CH2—CH2NH, 44%), 207 (13%), 209 (88%), 209 (22%); High resolution EI Calculated C20H19N2OF (M+.): 322.1481, Found: 322.1484 (100%).
To a solution of final compound 12 (82.3 mg, 0.25 mmol) in MeOH (10 mL) was added formaldehyde (96 μL, 1.22 mmol) and the mixture was hydrogenated (1 atm.) with 10% palladium-on-charcoal under vigorous stirring at room temperature for 1 hour. Then the mixture was filtered through a pad of celite and the solids were washed 4 times with CH2Cl2. After evaporation, the crude product was purified by column chromatograhy on silica gel using CHCl3/MeOH (3%) as eluent. Final compound 13 (43.4 mg, 50%) was obtained as a solid; mp: 139-141° C.
Mass spectrum: —CI m/z (assignment, relative intensity) 337 (MH+, 100%), 317 (MH+—HF, 30%), 279 (1%), 251 (1%), 209 (1%); EI m/z (assignment, relative intensity) 336 (M−, 74%), 293 (M+—COCH3, 13%), 265 (M+—CO═CHNHCH3, 9%), 233 (18%), 209 (42%), 196 (26%), 57 (100%); High resolution EICalculated C21H21N2OF (M+): 336.1638, Found: 336.1641 (100%).
To a solution of final compound 13 (34.3 mg, 0.1 mmol) in THF (10 mL) was added BH3.Me2S (100 μL, 0.2 mmol) and the mixture was heated at 85° C. overnight. Following evaporation of the solvent the residue was dissolved in 10 mL of HCl in MeOH (sat. solution) and the mixture was refluxed for 30 minutes. After removal of the solvent 10 mL of K2CO3 (sat. aq. solution) was added and the solution extracted 4 times with CH2Cl2. Then, the combined organic layers were evaporated and the crude product was purified by column chromatograhy on silica gel using CHCl3/MeOH (3%) as eluent. Final compound 14 (15.9 mg, 50%) was obtained as an oil.
Mass spectrum: —CI m/z (assignment, relative intensity) 323 (MH+, 73%), 303 (MH+—HF, 18%), 247 (4%), 219 (3%), 43 (100%); EI m/z (assignment, relative intensity 322 (M+, 73%), 278 (M+—N(CH3)2, 44%), 266 (M+—N(CH2)3, 85%), 264 (M+—CH2CH2NHCH3, 94%), 251 (M+—CH2CH2—CH2N(CH3), 100%), 209 (68%), 196 (38%); High resolution EI Calculated C21H23N2F (M+): 322.1845, Found: 322.1849 (100%).
To the above intermediate 25 (13.5 mg, 0.04 mmol) in CH2Cl2 (1 mL) was added CH3SO3H (1.3 μL, 0.02 mmol). After stiring at room temperature for 1 minute, the mixture was worked up by adding Na2CO3 (sat. aq. sol.). Extract 3 times with CH2Cl2 and dry with MgSO4. Column purification on silica gel using CHCl3/MeOH (95/05) gave final compound 15 as an oily product (10.5 mg, 82%).
Mass spectrum: —CI m/z (assignment, relative intensity) 309 (MH+, 100%), 289 (MH+—HF, 17%), 257(1%).
To a solution of imidazolone final compound 15 (10 mg, 0.03 mmol) in THF (1 mL) was added NaH (5 mg, 0.1 mmol) and the mixture was stirred at room temperature for 20 minutes. Then Me2SO4 (8 μL, 0.08 mmol) was added and the mixture was stirred for additional 30 min. Add 10 mL of NH4Cl (sat. aq. solution), extract with CH2Cl2 (3×10 mL) and dry with MgSO4. Column purification on silica gel using EtOAc/heptane (50/50) gave the N-methylated imidazolonefinal compound 16 (8.4 mg, 80%) as an oil.
Mass spectrum: —CI m/z (assignment, relative intensity) 323 (MH+, 100%), 303 (MH+—HF, 6%), 257 (11%), 252 (MH+—CH2N(CH3)CO, 9%), 229 (9%).
To a solution of intermediate 38 (0.17g, 0.5 mmol), CH2O (3 eq) and AcOH (3 eq) in MeOH (5 mL) at 0° C., NaCNBH3 (4 eq) was added in several lots. The reaction mixture was warmed to room temperature and stirred for 6 hours. Solid NaHCO3 (0.5 g) was added to the reaction mixture and stirred for 0.5 hour. To remove inorganic complexes the reaction mixture was put on sort filtration column and diluted with CH2Cl2:MeOH (9.5:0.5). The crude intermediate
thus obtained was dissolved in i-PrOH (4 mL) and a solution of KOH (56 mg) in water (0.5 mL) was added to it and then refluxed for 3 hours. Silica was added to the reaction mixture and the solvent was removed under reduced pressure followed by purification of compound by flash chromatography using CH2Cl2:MeOH (9:1) as an eluant to obtain final compound 17 as a thick viscous liquid (60%, 93 mg).
HRMS: Calculated 312.1638; found 312.1633.
a) To a solution of intermediate 39 (0.5 mmol, 0.33 g) in dioxane (5 mL) the corresponding amino alcohol (5 eq) was added and then refluxed for 6 hours. The solvent was removed under reduced pressure followed by chromatography (silica gel) using CH2Cl2:MeOH (9:1) as an eluant to obtain intermediates 39a, 39b and 39c as a thick viscous liquids in 40-50% overall yield.
b) A mixture of appropriate nosylamide intermediates 39a, 39b and 39c (ca. 0.4 mmol), thiophenol (110 mg, 1.0 mmol), anhydrous K2CO3 (138 mg, 1 mmol) and DMF (20 mL) was stirred at 80° C. for 4 hours, cooled to ambient temperature, diluted with water, product extracted with EtOAc (3×50 mL), combined organics washed with water (4×50 mL), brine (35 mL), dried (K2CO3), evaporated and purified by solid phase extraction on basic alumina (Brockmann II, heptane-ethyl acetate 50/50, then ethyl acetate-MeOH 100/0 to 96/4 to 90/10) to obtain final compounds 18, 19 and 20.
HRMS: Calculated 342.1744; found 342.1750.
HRMS: Calculated 397.2166; found 397.2158.
HRMS: Calculated 354.1744; found 354.1755.
To a solution of above intermediate 43 (81 mg, 0.25 mmol) in THF and water (3 mL/1 mL) was added Ph3P (0.13 g, 0.50 mmol). The reaction mixture was stirred at room temperature for 1 night. After evaporation of the solvent, MeOH (5 mL), HCHO (37 wt % aq. solution, 0.20 mL, 2.5 mmol), AcOH (1 mL) and NaCNBH3 (75 mg, 1.20 mmol) were added. Stirring was continued at room temperature for 1 day. Add Na2CO3 (sat. aq. sol.), extract 3 times with CH2Cl2. Column purification on silica gel using EtOAc gave final compound 21 as an oily product (70 mg, 86%).
Mass spectrum: —CI m/z (assignment, relative intensity) 328 (MH+, 100%), 308 (MH+—HF, 20%), 283 (MH+-Me2NH, 40%), 249 (MH+-Me2NH—H2S, 12%).
To a solution of above sulfone azide intermediate 44 (133.5 mg, 0.37 mmol) in THF (8 mL) was added water (67.0 μL, 3.74 mmol) and Ph3P (0.13 g, 0.50 mmol). The reaction mixture was stirred at room temperature for 1 night. After evaporation of the solvent, 5 mL of MeOH, HCHO (37 wt % aq. solution, 0.24 mL, 2.98 mmol), AcOH (0.5 mL) and NaCNBH3 (94 mg, 1.49 mmol) were added. Stirring was continued at room temperature for 1 day. Add Na2CO3 (sat. aq. sol.), extract 3 times with CH2Cl2. Column purification on silica gel using CH2Cl2/MeOH (95/05) gave final compound 22 as an oil product (60.7 mg, 45%).
Mass spectrum: —CI m/z (assignment, relative intensity) 360 (MH+, 100%), 358 (6%), 340 (MH+—HF, 12%), 303 (8%), 294 (MH+—H2SO2, 4%), 250 (1%).
To a solution of above intermediate 47 (0.15 g, 0.46 mmol) in THF and water (5mL/1 mL) was added Ph3P (0.13g, 0.50 mmol). After stirring at room temperature for 1 night and evaporation of the solvent, 5 mL of MeOH, HCHO (37 wt % aq. solution, 0.20 mL, 2.5 mmol), AcOH (1 mL) and NaCNBH3 (75 mg, 1.20 mmol) were added. Stirring was continued at room temperature for 1 day. Add Na2CO3 (sat. aq. sol.), extract 3 times with CH2C2. Column purification on silica gel using EtOAc gavefinal compound 23 as an oily product (70 mg, 86%).
Mass spectrum: —CI m/z (assignment, relative intensity) 328 (MH+, 100%), 308 (MH+ —HF, 20%), 283 (MH+-Me2NH, 40%), 249 (MH+-Me2NH—H2S, 12%).
To a solution of above sulfone azide intermediate 48 (146.5 mg, 0.41 mmol) in THF (8 mL) was added water (74.0 μL, 4.10 mmol) and Ph3P (0.215 mg, 0.82 mmol). The reaction mixture was stirred at room temperature for 1 night. After evaporation of the solvent, 5 mL of MeOH, HCHO (37 wt % aq. solution, 0.28 mL, 3.51 mmol), AcOH (0.5 mL) and NaCNBH3 (110.0 mg, 1.75 mmol) were added. Stirring was continued at room temperature for 1 day. Add Na2CO3 (sat. aq. sol.), extract 3 times with CH2Cl2. Column purification on silica gel using CH2Cl2/MeOH (90/10) gave final compound 24 as an oily product (105.0 mg, 71%).
Mass spectrum: —CI m/z (assignment, relative intensity) 360 (MH+, 100%), 358 (6%), 340 (MH+—HF, 12%), 303 (8%), 294 (MH+—H2SO2, 4%), 250 (1%).
To a solution of above intermediate 49 (107.9 mg, 0.32 mmol) in THF (5 mL) was added water (57 μL, 3.16 mmol) and Ph3P (166.0 mg, 0.63 mmol). The reaction mixture was stirred at room temperature for 1 night. After evaporation of the solvent 5 mL of MeOH, HCHO (37%, 0.26 mL, 3.33 mmol), AcOH (0.5 mL) and NaCNBH3 (104.7 mg, 1.67 mmol) were added. Stirring was continued at room temperature for 1 day. Add Na2CO3 (sat. aq. sol.), extract 3 times with CH2Cl2. Column purification on silica gel using CH2Cl2/MeOH (95/05) gave final compound 25 as an oily product (80.4 mg, 74%).
Mass spectrum: —CI m/z (assignment, relative intensity) 344 (MH+, 100%), 328 (MH+—O, 13%), 326 (MH+—H2O, 15%), 324 (MH+—HF, 15%), 182 (14%), 100 (27%).
To a solution of above intermediate 50 (133.4 mg, 0.39 mmol) in THF (5 mL) was added water (70 μL, 3.91 mmol) and Ph3P (205.2 mg, 0.78 mmol). The reaction mixture was stirred at room temperature for 1 night. After evaporation of the solvent, 5 mL of MeOH, HCHO (37%, 0.24 mL, 2.99 mmol), AcOH (0.4 mL) and NaCNBH3 (94.0 mg, 1.50 mmol) were added. Stirring was continued at room temperature for 1 day. Add Na2CO3 (sat. aq. sol.), extract 3 times with CH2Cl2. Column purification on silica gel using CH2Cl2/MeOH (95/05) gave final compound 26 as an oily product (85.2 mg, 63%).
Mass spectrum: —CI m/z (assignment, relative intensity) 344 (MH+, 100%), 328 (MH+—O, 10%), 327 (12%), 326 (MH+—H2O, 46%), 324 (MH+—HF, 22%), 283 (12%).
To a solution of intermediate 51 (85 mg, 0.25 mmol) in THF (5 mL) was added water (45 μL, 2.49 mmol) and Ph3P (130.8 mg, 0.50 mmol). The reaction mixture was stirred at room temperature for 1 night. After evaporation of the solvent, 5 mL of MeOH, HCHO (37%, 0.08 mL, 1.03 mmol), AcOH (0.3 mL) and NaCNBH3 (32 mg, 0.52 mmol) were added. Stirring was continued at room temperature for 1 day. Add Na2CO3 (sat. aq. sol.), extract 3 times with CH2Cl2. Column purification on silica gel using CH2Cl2/MeOH (95/05) gave final compound 27 as an oily product (35 mg, 41%).
Mass spectrum: —CI m/z (assignment, relative intensity) 344 (MH+, 100%), 328 (MH+—O, 4%), 327 (3%), 326 (MH+—H2O, 10%), 324 (MH+—HF, 8%), 281 (6%).
To a solution of above intermediate 52 (158.5 mg, 0.46 mmol) in THF (5 mL) was added water (84 μL, 4.65 mmol) and Ph3P (243.8 mg, 0.93 mmol). The reaction mixture was stirred at room temperature for 1 night. After evaporation of the solvent, 5 mL of MeOH, HCHO (37%, 0.32 mL, 4.05 mmol), AcOH (0.5 mL) and NaCNBH3 (130 mg, 2.03 mmol) were added. Stirring was continued at room temperature for 1 day. Add Na2CO3 (sat. aq. sol.), extract 3 times with CH2Cl2. Column purification on silica gel using CH2Cl2/MeOH (95/05) gave final compound 28 as an oily product (115.7 mg, 72%).
Mass spectrum: —CI m/z (assignment, relative intensity) 344 (MH+, 100%), 328 (MH+—O, 3%), 327 (3%), 326 (MH+—H2O, 13%), 324 (MH+—HF, 14%), 281 (6%).
Dissolve intermediate 23a (1.31 g, 2.63 mmol) in CH2Cl2 (50 mL). Add dihydropyran (1.20 mL, 13.2 mmol) and camphorsulfonic acid (6 mg, 0.026 mmol). Stir at room temperature for 5 hours. Evaporate the solvent and dissolve the residue in 50 mL MeOH. Add K2CO3 (0.73 g, 5.26 mmol) and stir at room temperature for 1 night. Work up by adding sat. aq. NH4Cl (30 mL), extract with CH2Cl2 (3×15 mL) and dry with MgSO4. Evaporate the solvent and dissolve the residue in dry THF (50 mL). Add NaH (0.24 g, 7.78 mmol) and stir at room temperature for 1 day. Add 30 mL sat. aq. NH4Cl, extract with CH2Cl2 (3×20 mL) and dry the organic phases with MgSO4. Column purification on silica gel using ether/hexane (35/65) gave an oil (0.86 g, 90% from 2). Dissolve this oil (0.86 g, 2.34 mmol) in 20 mL MeOH/H2O (9/1) and add Dowex 50WX8-100 (1.00 g). Heat the mixture at 50° C. for 1 night. Filter through a P3 filter, wash the solids with CH2Cl2 (5×15 mL) and evaporate the solvent. Column purification on silica gel using ether/hexane (70:30) yielded final compound 29 as an oil (0.61 g, 93%).
Mass spectrum: —CI m/z (assignment, relative intensity) 285 (MH+, 25%), 267 (MH+—H2O, 100%), 249 (MH−—2 H2O, 36%); EI: m/z (assignment, relative intensity) 284 (M−., 1%), 209 (M+. —CH2CHOHCH2OH, 100%); High resolution EI Calculated C18H17FO2 (M+.): 284.1213, Found: 284.1204 (2%).
Dissolve final compound 29 (0.61 g, 2.16 mmol) in CH2Cl2 (50 mL). Add Et3N (0.60 mL, 4.32 mmol), DMAP (0.13 g, 1.08 mmol) and MsCl (0.25 mL, 3.24 mmol). Stir at room temperature for 4 hours. Work up by adding sat. aq. NH4Cl (20 mL), extract with CH2Cl2 (3×20 mL) and dry with MgSO4. Column purification on silica gel using CH2Cl2 yielded intermediate 53 as an oil (0.76 g, 97%).
Mass spectrum: —CI m/z (assignment, relative intensity) 363 (MH+, 1%), 267 (MH−- MsOH, 100%), 249 (MH+-MsOH—H2O, 33%); EI: m/z (assignment, relative intensity) 362 (M+., 5%), 266 (M+.-MsOH, 3%), 248 (M+-MsOH—H2O, 4%), 209 (M−. —CH2CHOMsCH2OH, 100%); High resolution EI Calculated C19H19FO4S (M−.): 362.0988, Found: 362.0984 (12%).
Dissolve mesylate intermediate 53 (0.29 g, 0.79 mmol) in DMF (10 mL), add NaN3 (0.10 g, 1.58 mmol) and heat the mixture at 90° C. for 2 hours. Add sat. aq. NH4Cl (10 mL), extract with CH2Cl2 (3×10 mL) and dry with MgSO4. Column purification on silica gel using CH2Cl2/heptane (40:60) yielded intermediate 54 as a crystalline product (0.22 g, 88%); mp: 91-93° C.
Mass spectrum: —CI m/z (assignment, relative intensity) 310 (MH+, 13%), 282 (MH+—N2, 100%); EI: m/z (assignment, relative intensity) 281 (M+.—N2, 28%), 208 (100%); High resolution EI Calculated C18H16FNO (M+.—N2): 309.1216, Found: 309.1223 (40%).
Dissolve intermediate 54 (0.16 g, 0.52 mmol) in i-PrOH/THF (2:1, 15 mL). Add 10% Pd—C (ca. 100 mg) and subject to hydrogenation (1 atmospheric pressure) for 1 night. Filter through a pad of celite, wash the solids with CH2Cl2 (5×10 mL) and evaporate the filtrate. The residue is purified by column chromatography on silica gel using CHCl3/MeOH (75:25) to give final compound 30 as a crystalline product (0.14 g, 94%); mp:74-76° C.
Mass spectrum: —CI m/z (assignment, relative intensity) 284 (MH+, 100%); EI: m/z (assignment, relative intensity) 283 (M+., 5%), 209 (M+.—CH2CHNH2CH2OH, 100%) High resolution EI Calculated C18H18FNO (M+.): 283.1372, Found: 283.1370 (43%).
Dissolve final compound 29 (77 mg, 0.27 mmol) in CH2Cl2 (10 mL) and add pyridinium chlorochromate (131 mg, 0.54 mmol). Stir at room temperature for 20 hours. Filter through a pad of celite, wash the solids with CH2Cl2 (5×20 mL) and evaporate the filtrates. Column purification on silica gel using ether/hexane (50:50) yielded final compound 31 as a white crystalline product (61 mg, 80%); mp: 146-148° C.
Mass spectrum: —CI m/z (assignment, relative intensity) 283 (MH+, 11%), 265 (MH+—H2O, 100%), 237 (MH−—H2O—CO, 22%); EI: m/z (assignment, relative intensity) 282 (M+., 26%), 209 (M+.—CH2COCH2OH, 100%); High resolution EI Calculated C18H15FO2 (M+.): 282.1056, Found: 282.1057 (40%).
Intermediate 54 (0.24 g, 0.76 mmol) was dissolved in i-PrOH/THF (2:1, 15 mL). Add 10% Pd—C (ca. 150 mg) and subject the mixture to hydrogenation (1 atmospheric pressure) for 1 night. Add 35% aq. CH2O (0.60 mL, 7.6 mmol) and continue hydrogenation for 2 days. Filter through celite and wash with CH2Cl2 (5×15 mL). Combine the organic phases and dry with MgSO4. The solution was filtered and evaporated, and the residue was purified by column chromatography on silica gel using CHCl3/MeOH (90:10) to yield final compound 32 MH-170 as an oil (0.22 g, 93%).
Mass spectrum: —CI m/z (assignment, relative intensity) 312 (MH+, 100%); EI: m/z (assignment, relative intensity) 311 (M+., 7%).
Dissolve final compound 31 (0.18 g, 0.63 mmol) in i-PrOH/THF (2:1, 10 mL). Add 10% Pd—C (ca. 100 mg), Et3N (0.87 mL, 6.3 mmol) and MeNH2.HCl (0.42 g, 6.3 mmol). Subject the mixture to hydrogenation (1 atmospheric pressure) for 1 night. Filter through a pad of celite and wash the solids with CH2Cl2 (5×15 mL). The solution was filtered and evaporated and the residue was purified by column chromatography on silica gel using CHCl3/MeOH (90:10) to yield two diastereoisomers (0.18 g, 95%) with a ratio of 5:1, from which the major (3R)-isomer (final compound 33) can be partly separated.
Mass spectrum: —CI m/z (assignment, relative intensity) 298 (MH+, 100%); EI: m/z (assignment, relative intensity) 297 (M+., 5%), 266 (M+.—CH3NH2, 19%); High resolution EI Calculated C19H20FNO (M+.): 297.1529, Found: 297.1528 (3.5%).
a) Conversion of alkene into diastereoisomeric diols. Dissolve intermediate 56 (1.40 g, 4.52 mmol) in acetone (30 mL). Add a small crystal of OsO4 (catalytic amount) and N-methylmorpholine N-oxide (0.63 g, 5.42 mmol). Stir at room temperature for 1 day. The solvent was removed under reduced pressure, and the residue was purified by column chromatography on silica gel using EtOAc/hexane (80:20) to yield a mixture of two diastereoisomeric diols (oil, 1.45 g, 93%).
b) Selective mono-tosylation of primary alcohol group. Dissolve the above diols (1.45 g, 4.22 mmol) in toluene (50 mL). Add Et3N (1.76 mL, 12.6 mmol), TsCl (1.05 g, 5.48 mmol) and Bu2SnO (0.10 g, 0.42 mmol). Stir at room temperature for 1 day. Add sat. aq. NH4Cl (30 mL), extract with CH2Cl2 (3×20 mL) and dry with MgSO4. The solution was filtered and evaporated and the residue was purified by column chromatography using EtOAc/hexane (40:60) to yield the diastereoisomeric monotosylate derivatives corresponding to selective sulfonylation of the primary OH group (oil, 1.74 g, 83%).
c) Protection of secondary alcohol group. Dissolve the monotosylates (1.74 g, 3.49 mmol) in CH2Cl2 (60 mL) and add dihydropyran (1.59 mL, 17.5 mmol), camphorsulfonic acid (10 mg, 0.035 mmol). Stir at room temperature for 1 h and remove the solvent under reduced pressure.
d) Deprotection and cyclisation of benzylic alcohol. The residue was dissolved in MeOH (50 mL). Add K2CO3 (0.79 g, 6.99 mmol) and stir at room temperature for 1 night. Work up by adding sat. aq. NH4Cl (30 mL), extract 3 times with CH2Cl2 (3×20 mL) and dry with MgSO4. The solvent was evaporated and the residue containing the benzylic alcohol was dissolved in dry THF (50 mL). Add NaH (0.21 g, 6.99 mmol) and stir at room temperature for 3 days to effect cyclisation. Work it up by adding sat. aq NH4Cl (30 mL) and extract with CH2Cl2 (3×20 mL). Dry with MgSO4 and evaporate the solvent.
e) Deprotection and oxidation of secondary alcohol group. Dissolve the residue (1.70 g) in 20 mL MeOH/H2O (9:1) and add Dowex 50WX8-100 (1.00 g). Heat the mixture at 50° C. for 2 hours. Filter through P3 filter, wash the solids with CH2Cl2 (5×15 mL) and evaporate. Column purification on silica gel using ether/hexane (70:30) yielded an oil (two diastereoisomeric alcohols) (0.87 g, 88%).
Dissolve the above oil (0.87 g, 3.06 mmol) in CH2Cl2 (40 mL). Add pyridinium chlorochromate (1.32 g, 6.13 mmol) and stir at room temperature for 1 night. Filter through a pad of celite, wash the solids with CH2Cl2 (5×20 mL) and evaporate. Column purification on silica gel using CH2Cl2/hexane (80:20) yielded final compound 34 as an oil (0.66 g, 76%).
Mass spectrum: —CI m/z (assignment, relative intensity) 283 (MH+, 25%), 265 (MH+—H2O, 100%); EI: m/z (assignment, relative intensity) 282 (M−., 39%), 209 (M+.—CH2COCH2OH, 100%).
Dissolve final compound 34 (0.23 g, 0.83 mmol) in i-PrOH (15 mL). Add Et3N (1.15 mL, 8.25 mmol), MeNH2HCl (0.56 g, 8.25 mmol) and 10% Pd/C (ca. 150 mg). Subject to hydrogenation (1 atmospheric pressure) for 1 night. Filter through a pad of celite and wash the solids with CH2Cl2 (5×10 mL). The solution was filtered and evaporated, and the residue was purified by column chromatography on silica gel using CHCl3/MeOH (90:10) to yield final compound 35 as the nearly exclusive diastereoisomer (0.23 g, 95%).
Mass spectrum: —CI m/z (assignment, relative intensity) 298 (MH+, 100%); EI: m/z (assignment, relative intensity) 209 (M+.—CH2CH(NHMe)CH2OH, 100%).
Representative Procedure—Synthesis of Methyl (2R,3aR,12bS)-2-(Aminomethyl)-11-fluoro-3,3a,8,12b-tetrahydrodibenzo[3,4:6,7]cyclohepta[1,2-b]pyrrole-1(2H)-carboxylate (intermediate 62b): A solution of triphenylphosphine (996 mg, 3.8 mmol) in dry THF (20 mL) was placed in two-necked 100 mL flask, equipped with septum, argon inlet and magnetic stirrer; cooled down to −15° C. Neat diisopropyl azodicarboxylate (768 mg, 3.8 mmol) was added through a septum with intensive stirring. Resulting yellow suspension was stirred at above temperature for 30 minutes, then carbamate intermediate 62 (650 mg, 1.9 mmol) in THF (5 mL) was added in one portion. After 5 minutes of stirring, diphenylphosphoryl azide (606 mg, 2.2 mmol) in THF (3 mL) was added dropwise for 3 minutes, resulting turbid mixture allowed to warm up to room temperature and stirred then for 12 hours. After this time water (0.2 mL) and triphenylphosphine (996 mg, 3.8 mmol) was added, and solution stirred at 45° C. for 2 hours. After cooling down to room temperature, silica gel (Kieselgel 60, 70-230 mesh, 4 g) was added, THF removed in vacuo, and silica powder submitted to the flash chromatography (Kieselgel 60, 230-400 mesh, CH2Cl2-MeOH, 100/0, gradually to 85/15) to give desired amine intermediate 62b (401 mg, 1.18 mmol, 62%) as colorless oil, darkening on standing.
Intermediate 62b
HRMS Calcd. for C20H21FN2O2: 340.1587; Found: 340.1588.
Intermediate 62c: two rotamers present (ca. 2:1 ratio)
CI-MS (CH4) 325 (MH+, 100%); 305 (MH+—HF, 10%).
HRMS Calcd. for C20H21FN2O: 324.1638; Found: 324.1644.
Intermediate 62d: two rotamers present (ca. 5:2 ratio)
HRMS Calcd. for C19H19FN2O: 310.1481; Found: 310.1480.
Amine intermediate 62b (401 mg, 1.18 mmol) was dissolved in MeOH (30 mL), AcOH (1 mL) and 35% aqueous formaldehyde (1 g, 11.7 mmol) added, followed by sodium cyanoborohydride (628 mg, 10 mmol). The resulting mixture was stirred at room temperature for 4 hours, quenched with concentrated HCl (5 mL), treated with solid NaHCO3 (8.4 g, 100 mmol), 1N sodium hydroxide (15 mL). The precipitated product was filtered off, washed with water (5×25 mL), dissolved in ethyl acetate, washed with brine (30 mL), dried (K2CO3), evaporated in vacuo and purified by column chromatography (Kieselgel 60, 230-400 mesh, CH2Cl2-MeOH 95/5 to 90/10 to 85/15) to give final compound 36a (313 mg, 0.85 mmol, 72%) as yellowish oil.
Final compound 36a:
HRMS Calcd. for C22H25FN2O2: 368.1900; Found: 368.1895.
Final compound 36b: Two rotamers, ca. 3:2 ratio.
HRMS Calcd. for C22H25FN2O: 352.1951; Found: 352.1955.
Final compound 36c: Two rotamers, ca. 5:3 ratio.
HRMS Calcd. for C21H23FN2O 338.1794; Found: 338.1790.
A mixture of final compound 36a (100 mg, 0.27 mmol), i-PrOH (10 mL), potassium hydroxide (560 mg, 10 mmol) and water (0.1 mL) was refluxed under nitrogen atmosphere for 12 hours (oil bath temperature 135° C.), then cooled to room temperature. After dilution with water (50 mL), extraction with EtOAc (3×40 mL), the combined organics were washed with water (3×40 mL), brine (40 mL), dried over K2CO3 and evaporated to give pure final compound 37 (84 mg, 100%) as yellowish semisolid, which was converted to the hydrochloride salt (final compound 37a).
HRMS Calcd. for C20H23FN2: 310.1845; Found: 310.1851.
A mixture of aldehyde intermediate 63 (50 mg, 0.162 mmol), methylamine hydrochloride (218 mg, 3.24 mmol), Et3N (405 mg, 4.0 mmol), 10% Pd—C (30 mg) and MeOH (12 mL) was hydrogenated for 2 hours at atmospheric pressure. The reaction mixture was filtered through Kieselguhr, which was subsequently washed with EtOAc (2×10 mL). The combined solutions were evaporated in vacuo and residue was purified by column chromatography (Kieselgel 60, 70-230 mesh, CH2Cl2/MeOH 100/0 to 85/15) to give final compound 38 (21 mg, 0.065 mmol, 40%) as brown oil; Four rotamers present (10:6:4:1 ratio).
CI-MS (CH4): 325 (100%, M+H−), 305 (12%, —HF). HRMS Calcd. for C20H21FN2O: 324.1638; Found: 324.1650.
Hydroxyacetaldehyde dimer (2,5-dihydroxy-1,4-dioxane) (240 mg, 2.0 mmol) was dissolved in MeOH (25 mL) and stirred at 40° C. for 30 minutes, then amine compound 37 (124 mg, 0.40 mmol) was added and stirring at 40° C. continued for another 30 minutes. After cooling down to room temperature, AcOH (120 mg, 2.0 mmol) was added, followed by sodium cyanoborohydride (188 mg, 3.0 mmol) and the resulting mixture was stirred for 2 hours. After this time it was quenched with concentrated HCl (2 mL), treated with solid NaHCO3 (2.94 g, 35 mmol), 1N sodium hydroxide (3 mL). About 20 mL of MeOH was removed in vacuo, the residue diluted with water (30 mL), and extracted with EtOAc (3×30 mL). The combined organics were washed with water (5×25 mL), brine (30 mL), dried (K2CO3), evaporated in vacuo and purified by column chromatography (Kieselgel 60, 230-400 mesh, CH2Cl2-MeOH 95/5 to 90/10 to 85/15) to give amine compound 39 (80 mg, 0.244 mmol, 61%) as colorless oil.
HRMS Calcd. for C22H27FN2O: 354.2107; Found: 354.2107.
2-((2R,3 aR,12bS)-2-[(Dimethylamino)methyl]-11-fluoro-3,3a,8,12b-tetrahydrodibenzo[3,4:6,7]cyclohepta[1,2-b]pyrrol-1(2H)-yl)ethanol compound 39 (50 mg, 0.153 mmol) was dissolved in dry THF (10 mL), then 60% NaH dispersion (8 mg, 0.2 mmol) was added, followed by dimethyl sulfate (25 mg, 0.2 mmol). The resulting mixture was stirred under argon atmosphere at 60° C. for 5 hours, then cooled, quenched with concentrated ammonium hydroxide (2 mL), diluted with water (40 mL). After extraction of product with EtOAc (3×25 mL) the combined organics were washed with water (3×25 mL), brine (25 mL), dried over K2CO3, evaporated in vacuo, and the residue purified by column chromatography (Kieselgel 60, 230-400 mesh, CH2Cl2-MeOH 95/5 to 90/10 to 85/15) to give final compound 40 (39 mg, 0.107 mmol, 70%) as yellowish oil.
HRMS Calcd. for C23H29FN2O: 368.2264; Found: 368.2270.
Poly(4-vinylpyridine) crosslinked with 2% divinylbenzene (0.5 g) was swollen for 1 hour with CH2Cl2 (10 mL), thenfinal compound 37 (57 mg, 0.184 mmol) in CH2Cl2 (2 mL) was added in one portion, followed by cyanogen bromide (39 mg, 0.367 mmol), then suspension stirred at room temperature for 30 minutes. The resin was filtered off, filtrate treated with saturated aqueous K2CO3 (10 mL), organic phase was separated, evaporated in vacuo, and the residue purified by column chromatography (Kieselgel 60, 230-400 mesh, CH2Cl2-MeOH 95/5 to 90/10→87/13) to give final compound 41 (24 mg, 0.077 mmol, 42%) as brownish oil.
CI-MS (CH4): 308 (100%, M+H+), 288 (8%, —HF).
HRMS Calcd. for C19H18FN3: 307.1485; Found: 307.1499.
To a solution of the aziridine intermediate 64 (63 mg, 0.237 mmol) in acetonitrile (1 mL) was added sodium iodide (107 mg, 0.711 mmol) and trimethylsilyl chloride (90 μL, 0.711 mmol) at room temperature. After the solution was stirred for 2 hours, morpholine (44 mg, 0.5 mmol) in acetonitrile (0.5 mL) was added dropwise to the mixture. The solution was heated to the boiling point of the solvent for 2 hours. The dark brown reaction mixture was quenched with aqueous 1.2N HCl solution and then was treated with sat. NaHCO3. The organic layer was separated and the aqueous layer was extracted with methylene chloride (3×10 mL). The combined organic extracts were washed with 20 mL of brine, dried over anhydrous MgSO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on basic alumina (Brockmann III, EtOAc-MeOH, 100/0 to 98/2 to 95/5) gave final compound 42 (3 8 mg, 0.11 mmol, 45%) as brownish oil.
CI-MS (CH4): 353 (100%, M+H−); 333 (—HF, 7%).
HRMS Calcd. for C22H25FN2O: 352.1951; Found: 352.1966.
A mixture of appropriate nosylamide intermediate 66a or 66b (ca. 0.4 mmol), thiophenol (110 mg, 1.0 mmol), anhydrous K2CO3 (138 mg, 1 mmol) and DMF (20 mL) was stirred at 80° C. for 4 hours, cooled to ambient temperature, diluted with water, product extracted with EtOAc (3×50 mL), combined organics washed with water (4×50 mL), brine (35 mL), dried (K2CO3), evaporated and purified by solid phase extraction on basic alumina (Brockmann II, heptane-ethyl acetate 50/50, then EtOAc-MeOH 100/0 to 96/4 to 90/10) to give compound 43a (111 mg, 0.28 mmol, 52% from intermediate 65) or compound 43b (80 mg, 0.24 mmol, 44% from intermediate 65), both as brownish oils.
Compound 43a (TK-895):
HRMS: Calcd. for C24H30FN3O: 395.2373; Found: 395.2374.
Compound 43b (TK-1013):
HRMS Calcd. for C21H25FN2O: 340.1951; 340.1943.
Tosylate intermediate 67k (153 mg, 0.338 mmol), 40% aqueous methylamine (15 mL), and THF (35 mL) were heated in stainless-steel bomb at 135° C. for 15 hours. After cooling, the bomb was opened, THF and methylamine evaporated in vacuo, residue extracted with CH2Cl2 (4×20 mL). The combined organics were washed with water (3×20 mL), dried (K2CO3), evaporated and purified by column chromatography (Kieselgel 60, 230-400 mesh, CH2Cl2-MeOH 98/2 to 85/15) to afford final compound 44 (32 mg, 0.098 mmol, 29%) as a brown oil, which was converted to the oxalate salt (final compound 44a).
HRMS Calcd. for C21H25FN2: 324.2002; Found: 324.1995.
final compound 45
Conversion of final compound 44 (48 mg, 0.15 mmol) with methyl chloroformate was carried out in the same way as described for the preparation of intermediate 62 Column chromatography (Kieselgel 60, 70-230 mesh, MeOH—CH2Cl2 3/97 to 15/85) afforded final compound 45 (45 mg, 0.118 mmol, 79%) as colorless oil.
HRMS Calcd. for C23H27FN2O2: 382.2057; Found: 382.2064.
Tosylate intermediate 68e (282 mg, 0.62 mmol), 40% aqueous methylamine (25 mL), and THF (35 mL) were heated in a steel bomb at 135° C. for 15 hours. After cooling, bomb was opened, THF and methylamine evaporated in vacuo. The residue was extracted with CH2Cl2 (4×30 mL) and the combined organics were washed with water (3×20 mL), dried (K2CO3) and evaporated. Crystallization from CH2Cl2/hexane gave final compound 46 (70 mg, 0.225 mmol, 36%) as beige powder.
HRMS Calcd. for C20H22FNO: 311.1685; Found: 311.1700.
Azide intermediate 69i (122 mg, 0.39 mmol) was dissolved in MeOH (10 mL), 10% palladium on carbon (40 mg) was added and the mixture submitted to the hydrogenation under atmospheric pressure for 1.5 hour, then 35% aqueous formaldehyde (1 g) and AcOH (120 mg, 2 mmol) were added, and hydrogenation continued for 2 hours. After filtration through short pad of Celite, and addition of EtOAc (45 mL), the reaction mixture was washed with saturated aqueous sodium bicarbonate (25 mL), water (2×50 mL), brine (30 mL), dried over K2CO3 and evaporated in vacuo. The residue was purified by column chromatography (Kieselgel 60, 70-230 mesh, ethyl acetate-MeOH, 100/0 to 95/5 to 92/8 to 87/13) to afford final compound 47 (77 mg, 0.248 mmol, 63%) as yellow oil. Product is a mixture of 2 epimers (12.8:1 ratio).
HRMS Calcd. for C20H22FNO: 311.1685; Found: 311.1680.
CI-MS (CH4) 312 (MH+, 100%); 292 (MH+—HF, 9%).
Reaction of intermediate 70d (100 mg, 0.304 mmol) was carried out was carried out in the same way as described for final compound 47. Purification by solid phase extraction (Alltech C18 2 g cartridge, wter-MeOH, 100/0 to 50/50 to 0/100) furnished compound 48 (57 mg, 0.18 mmol, 59%). Product is a mixture of 2 epimers (2:1 ratio).
HRMS Calcd. for C20H22FNO: 311.1685; Found: 311.1692.
CI-MS (CH4) 312 (MH+, 100%); 292 (MH+—HF, 12%).
A mixture of intermediate 71f (194 mg, 0.53 mmol) and 10% palladium on carbon (50 mg) in MeOH (35 mL) was hydrogenated at atmospheric pressure for 40 minutes, then 35% aqueous formaldehyde (1 mL) was added and hydrogenation continued for another 40 minutes. After filtration through short pad of Celite, reaction mixture was evaporated in vacuo. The residue was dissolved i-PrOH (20 mL), KOH (560 mg, 10 mmol) and water (0.1 mL) were added and resulting solution was refluxed under nitrogen atmosphere for 12 hours (oil bath temperature 135° C.), then cooled to room temperature. After dilution with water (50 mL), extraction with EtOAc (3×40 mL), the combined organics were washed with water (3×40 mL), brine (40 mL), dried over K2CO3 and evaporated in vacuo. The residue was purified by column chromatography (basic alumina, Brockmann activity 1, ethyl acetate-MeOH, 100/0 to 85/15) to give purefinal compound 49 (102 mg, 0.33 mmol, 62%) as brown oil. Product is a mixture of 2 epimers (1:1 ratio)
HRMS Calcd. for C20H23FN2: 310.1845; Found: 310.1833.
Tables 1-3 list compounds of Formula (I), which were prepared according to one of the above examples.
The interaction of the compounds of Formula (I) with 5-HT2A and 5-HT2C receptors was assessed in in vitro radioligand binding experiments. In general, a low concentration of a radioligand with a high binding affinity for the receptor is incubated with a sample of a tissue preparation enriched in a particular receptor (1 to 5 mg tissue) in a buffered medium (0.2 to 5 ml). During the incubation, the radioligands bind to the receptor. When equilibrium of binding is reached, the receptor bound radioactivity is separated from the non-bound radioactivity, and the receptor bound activity is counted. The interaction of the test compounds with the receptors is assessed in competition binding experiments. Various concentrations of the test compound are added to the incubation mixture containing the tissue preparation and the radioligand. Binding of the radioligand will be inhibited by the test compound in proportion to its binding affinity and its concentration. The affinities of the compounds for the 5-HT2 receptors were measured by means of radioligand binding studies conducted with: (a) human cloned 5-HT2A receptor, expressed in L929 cells using [125I]R91150 as radioligand and (b) human cloned 5-HT2C receptor, expressed in CHO cells using [3H]mesulergine as radioligand.
Cortex from rat brain was collected and homogenised using an Ultra-Turrax T25 and a Dual homogeniser in ice-cold homogenising buffer containing Tris, NaCl and KCl (50 mM, 120 mM and 5 mM, respectively, pH 7.4) prior to dilution to an appropriate protein concentration optimised for specific and non-specific binding. Binding was performed with radioligand [3H]Nixosetine (NEN, NET-1084, specific activity ˜70 Ci/mmol) diluted in ice cold assay buffer containing Tris, NaCl and KCl (50 mM, 300 mM and 5 mM, respectively, pH 7.4). at a concentration of 20 nmol/L. Prepared radioligand (50 μl) was then incubated (60 min, 25° C.) with membrane preparations pre-diluted to an appropriate protein concentration (400 μl), and with 50 μl of either the 10% DMSO control, Mazindol (10−6 mol/L final concentration), or compound of interest. Membrane-bound activity was detected by filtration through a Packard Filtermate harvester onto GF/B Unifilterplates, washed with ice-cold Tris-HCl buffer, containing NaCl and KCl (50 mM, 120 mM and 4 mM; pH 7.4; 6×0.5 ml). Filters were allowed to dry for 24 h before adding scintillation fluid. Scintillation fluid was allowed to saturate filters for 24 h before counting in a Topcount scintillation counter. Percentage specific bound and competition binding curves were calculated using S-Plus software (Insightful).
Frozen membranes of human Dopamine D2L receptor-transfected CHO cells were thawed, briefly homogenised using an Ultra-Turrax T25 homogeniser and diluted in Tris-HCl assay buffer containing NaCl, CaCl2, MgCl2, KCl (50, 120, 2, 1, and 5 mM respectively, adjusted to pH 7.7 with HCl) to an appropriate protein concentration optimised for specific and non-specific binding. Radioligand [3H]Spiperone (NEN, specific activity ˜70 Ci/mmol) was diluted in assay buffer at a concentration of 2 nmol/L. Prepared radioligand (50 μl), along with 50 μl of either the 10% DMSO control, Butaclamol (10−6 mol/l final concentration), or compound of interest, was then incubated (30 min, 37° C.) with 400 μl of the prepared membrane solution. Membrane-bound activity was filtered through a Packard Filtermate harvester onto GF/B Unifilterplates and washed with ice-cold Tris-HCl buffer (50 mM; pH 7.7; 6×0.5 ml). Filters were allowed to dry before adding scintillation fluid and counting in a Topcount scintillation counter. Percentage specific bound and competition binding curves were calculated using S-Plus software (Insightful).
“Active ingredient” (A.I.) as used throughout these examples relates to a compound of Formula (I), a pharmaceutically acceptable acid addition salt, a stereochemically isomeric form thereof or a N-oxide form thereof.
Methyl 4-hydroxybenzoate (9 g) and propyl 4-hydroxybenzoate (1 g) were dissolved in boiling purified water (4 l). In 3 l of this solution were dissolved first 2,3-dihydroxybutanedioic acid ( 10 g) and thereafter A.1 (20 g). The latter solution was combined with the remaining part of the former solution and 1,2,3-propanetriol (12 l) and sorbitol 70% solution (3 l) were added thereto. Sodium saccharin (40 g) were dissolved in water (500 ml) and raspberry (2 ml) and gooseberry essence (2 ml) were added. The latter solution was combined with the former, water was added q.s. to a volume of 20 l providing an oral solution comprising 5 mg of the active ingredient per teaspoonful (5 ml). The resulting solution was filled in suitable containers.
Preparation of Tablet Core
A mixture of A.I. (100 g), lactose (570 g) and starch (200 g) was mixed well and thereafter humidified with a solution of sodium dodecyl sulfate (5 g) and polyvinylpyrrolidone (10 g) in water (200 ml). The wet powder mixture was sieved, dried and sieved again. Then there was added microcrystalline cellulose (100 g) and hydrogenated vegetable oil (15 g). The whole was mixed well and compressed into tablets, giving 10.000 tablets, each containing 10 mg of the active ingredient.
Coating
To a solution of methyl cellulose (10 g) in denaturated ethanol (75 ml) there was added a solution of ethyl cellulose (5 g) in dichloromethane (150 ml). Then there were added dichloromethane (75 ml) and 1,2,3-propanetriol (2.5 ml). Polyethylene glycol (10 g) was molten and dissolved in dichloromethane (75 ml). The latter solution was added to the former and then there were added magnesium octadecanoate (2.5 g), polyvinylpyrrolidone (5 g) and concentrated colour suspension (30 ml) and the whole was homogenated. The tablet cores were coated with the thus obtained mixture in a coating apparatus.
Methyl 4-hydroxybenzoate (1.8 g) and propyl 4-hydroxybenzoate (0.2 g) were dissolved in boiling water (500 ml) for injection. After cooling to about 50° C. there were added while stirring lactic acid (4 g), propylene glycol (0.05 g) and A.I. (4 g). The solution was cooled to room temperature and supplemented with water for injection q.s. ad 1000 ml, giving a solution comprising 4 mg/ml of A.I. The solution was sterilized by filtration and filled in sterile containers.
| Number | Date | Country | Kind |
|---|---|---|---|
| PCT/EP04/106373 | Dec 2004 | WO | international |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/EP2005/056544 | 12/6/2005 | WO | 00 | 8/7/2007 |
| Publishing Document | Publishing Date | Country | Kind |
|---|---|---|---|
| WO2006/061342 | 6/15/2006 | WO | A |
| Number | Name | Date | Kind |
|---|---|---|---|
| 20090023721 | Megens et al. | Jan 2009 | A1 |
| Number | Date | Country |
|---|---|---|
| 9738991 | Oct 1997 | WO |
| 9919317 | Apr 1999 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 20080287450 A1 | Nov 2008 | US |
