WO2004111056A2 - Compound libraries of spiro-benzodioxine-piperidines - Google Patents

Compound libraries of spiro-benzodioxine-piperidines Download PDF

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Publication number
WO2004111056A2
WO2004111056A2 PCT/GB2004/002533 GB2004002533W WO2004111056A2 WO 2004111056 A2 WO2004111056 A2 WO 2004111056A2 GB 2004002533 W GB2004002533 W GB 2004002533W WO 2004111056 A2 WO2004111056 A2 WO 2004111056A2
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list
cont
compounds
library
spiro
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PCT/GB2004/002533
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French (fr)
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WO2004111056A3 (en
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Emma Barker
Andrew Proud
Martin Slater
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Biofocus Plc
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/10—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
    • C40B50/08—Liquid phase synthesis, i.e. wherein all library building blocks are in liquid phase or in solution during library creation; Particular methods of cleavage from the liquid support

Definitions

  • the present invention relates to compounds capable of binding to G-protein coupled receptors.
  • libraries of compounds are provided for use in screening programmes against GPCR targets as well ' as the individual compounds for use in hit to lead and lead optimisation projects and similar stages in the drug discovery process .
  • the method also provides methods for making compounds and libraries .
  • these collections will be drug-like but are likely to be limited in scope and will be directed to certain areas of a particular project. It has been the common practice of many pharmaceutical companies in recent times to augment the collections by purchasing either single compounds from vendors or by contracting the synthesis of combinatorial libraries of compounds. The singly purchased compounds may have been selected to fill in areas of compound space poorly represented in the compound collections . Combinatorial libraries are typically synthesised around well- performing chemistries with some design based on producing 'diversity' in compound space.
  • a complementary approach, and one that is increasingly preferred, is to screen focused libraries against the target of choice. Focused libraries are becoming of increasing importance in their ability to generate hits capable of rapid expansion in many areas including GPCRs . Such libraries are slightly more expensive to prepare but have attributes of reliability, reproducibility and provide a considerably higher hit rate: typically 10-100 fold and above compared with random screening. They are, however, very difficult to design and their efficiency relates directly to the amount of effort that has gone into the design. Using focused libraries, it is usually possible to get a number of hits in the low micromolar and below range. As there is a defined set of compounds there is the potential to observe indications of SAR in a chemical series and progress the chemistry efficiently.
  • GPCRs G-protein-coupled receptors
  • the rhodopsin receptor is somewhat unusual in its interactions with its ligand and is not used as a drug target. Nevertheless, the overall three dimensional arrangement can be deduced from the X-ray and is in accordance with previous work based upon bacteriorhodopsin receptor which is not G-protein-coupled.
  • GPCRs are most often characterised by sequence homology as being comprised of several sub-families. Most attention currently is directed towards Family A receptors as being the most tractable class historically and also the one with the most potential targets .
  • Family A comprises about 300 receptors that are potential drug targets, approximately half of which have known ligands and the rest, the so-called orphan receptors.
  • the group of druggable receptors is composed essentially of two types: those whose natural ligand interacts wholly within the transmembrane domain, such as the aminergic, nucleotide-like, prostaglandin receptors, etc. and those peptide liganded receptors, which have a large part of their interactions in the extracellular region and which may insert a peptide loop or tail into the transmembrane region to effect signal transduction. Examples of this class are angiotensin, cholecystokinin and opioid receptors.
  • the focused library provided herein is designed to interact with a range of the family A receptors .
  • Each library is a defined set of compounds which will enhance the probability of finding a small molecule which will interact with one or more type of GPCR receptor.
  • focused libraries can be provided having compounds which will interact with aminergic GPCRs, and peptidic GPCRs requiring an obligatory positive charge in ligands, or other types or groups of GPCRs .
  • Focused libraries according to this invention can provide hit rates of 1-13% or more for the requisite predicted GPCRs from both amine- and peptide-liganded classes and with agonists and antagonists .
  • library means a group of compounds which are structurally related by virtue of a core chemical structure (or “scaffold”) but which differ from each other by virtue of permutation of specific substituent groups attached to the scaffold.
  • such a library will consist of or comprise a number of compounds, e.g. as many as about 100, 1000,2000, 3000 or indeed 10,000 compounds.
  • the number of compounds should be sufficient to provide an adequate diversity of related compounds without being so large as to be unduly complex/expensive to produce .
  • the substituent may appear in the compound exactly as shown (i.e. simply covalently bonded to the scaffold) or may be a derivative of the shown chemical formula of the substituent by virtue of use of a reactive group to couple the substituent to the scaffold.
  • the total number of permutations created by the permitted substituents may be a very large number, far greater in magnitude than the actual number of compounds in an actual library.
  • the number of possible compounds for any "virtual" library may well greatly exceed the number of synthesised compounds making up an embodiment of the "real" library.
  • the invention is intended to encompass libraries having all, and a number, which is less than all, of the permitted substitutions represented by compounds therein.
  • the present invention provides novel focused libraries of compounds .
  • Most of the compounds defined by the permitted substitutions on the scaffolds are also novel compounds per se and the invention is intended to encompass each individual novel compound.
  • Any known compound having a structural formula identical to any one of the compounds covered by the formulae of scaffolds and permitted substitutions described herein is hereby explicitly disclaimed per se.
  • SFG07 is designed to target a large group of receptors that recognise electron rich aromatic ring systems. These receptors fall into two main groups, those that also require a positively charged amine in the ligand (for example the adrenoceptors or the serotonin receptors) and those that do not (for example the melanocortin, vasopressin, and melatonin receptors) .
  • a positively charged amine in the ligand for example the adrenoceptors or the serotonin receptors
  • melanocortin, vasopressin, and melatonin receptors for example the melanocortin, vasopressin, and melatonin receptors
  • the invention provides a compound comprising or consisting of a set of structurally related compounds of general formula (I) :
  • the 4H-spiro [1, 3-benzodioxine-2, 4 ' -piperidine] (IV) can be prepared from an appropriately substituted 2-hydroxybenzyl alcohol (II) , selected from List 1, by reaction with a suitably N-protected 4-piperidinone (III) followed by appropriate deprotection. Suitable protecting groups would include ethoxy carbonyl, benzoxy carbonyl, acetyl, benzyl.
  • the 4H- spiro [1,3 -benzodioxine-2, 4 ' -piperidine] (IV) can be further derivatised by modification of the Rl group.
  • the Rl group may be modified by a number of reactions, including so called Suzuki reactions and other palladium catalysed reactions.
  • the Gilson 215 was used as both auto-sampler and fraction collector
  • the gradient used was 90% water (5 mM ⁇ H 4 HC0 3 ) / 10% methanol for 1.5 min to 100% methanol over 6.5 min then held at 100% methanol for 3.0 min. The solvent mixture was then returned to the initial conditions over 0.5 min.
  • the purification was controlled by Unipoint software, triggering a threshold collection value monitoring at 220 nm. Collected fractions were analysed by LCMS (Waters/Micromass ZQ) (Table 1) . The fractions that contained the desired product are concentrated by vacuum centrifugation and the resultant residue dried by freeze-drying .

Abstract

The present invention provides a compound library designed to target a large group of receptors that recognise electron rich aromatic ring systems. These receptors fall into two main groups, those that also require a positively charged amine in the ligand (for example the adrenoceptors or the serotonin receptors) and those that do not (for example the melanocortin, vasopressin, and melatonin receptors). A combination of specific motifs R1 and R2 are appeneded form the central scaffold and are designed to pick up different interactions at the receptor site. The library comprises or consists of a set of structurally related compounds of general formula (I).

Description

COMPOUND LIBRARIES
Introduction
Background
The present invention relates to compounds capable of binding to G-protein coupled receptors. In particular, libraries of compounds are provided for use in screening programmes against GPCR targets as well 'as the individual compounds for use in hit to lead and lead optimisation projects and similar stages in the drug discovery process .
The method also provides methods for making compounds and libraries .
As part of the process of discovering drugs or agrochemicals it is customary to screen libraries of compounds against biological targets to discover "Hits' which are then further developed into 'Leads' and subsequently drugs or agrochemicals by using the techniques of medicinal chemistry. Accordingly the success or not of a drug or agrochemical discovery project is critically dependent on the quality of the hit and this in turn is dictated by the quality of the screening library.
Technological advances have enabled screening on a very large scale and the screening of hundreds of thousands of compounds at the start of a discovery program is routine. This, however, does entail a significant cost. The hits obtained from such screening efforts are not all of the best quality and often take a large amount of subsequent time and effort in order to get a good lead. It has been estimated that only about 25% of projects actually get to the lead optimisation stage and part -of the reason for this is the intractability of hits from high throughput screening. Screening libraries are commonly collections of compounds from several sources. As a result, they typically contain compounds synthesised as a part of previous projects in the history of a company. With regard to drug discovery, these collections will be drug-like but are likely to be limited in scope and will be directed to certain areas of a particular project. It has been the common practice of many pharmaceutical companies in recent times to augment the collections by purchasing either single compounds from vendors or by contracting the synthesis of combinatorial libraries of compounds. The singly purchased compounds may have been selected to fill in areas of compound space poorly represented in the compound collections . Combinatorial libraries are typically synthesised around well- performing chemistries with some design based on producing 'diversity' in compound space.
A complementary approach, and one that is increasingly preferred, is to screen focused libraries against the target of choice. Focused libraries are becoming of increasing importance in their ability to generate hits capable of rapid expansion in many areas including GPCRs . Such libraries are slightly more expensive to prepare but have attributes of reliability, reproducibility and provide a considerably higher hit rate: typically 10-100 fold and above compared with random screening. They are, however, very difficult to design and their efficiency relates directly to the amount of effort that has gone into the design. Using focused libraries, it is usually possible to get a number of hits in the low micromolar and below range. As there is a defined set of compounds there is the potential to observe indications of SAR in a chemical series and progress the chemistry efficiently.
G-protein-coupled receptors (GPCRs) are very important in the regulation of numerous body processes and a significant proportion of all drugs work by interaction with these receptors. There are several hundred known, many of which are orphans - those receptors that have no established ligands. They fall into a class of 7-transmembrane receptors and there is only one X-ray structure known, that of the bovine rhodopsin receptor, and this is at a resolution of 2.8 Angstroms and is thus not suitable for accurate modeling work. In addition, the rhodopsin receptor is somewhat unusual in its interactions with its ligand and is not used as a drug target. Nevertheless, the overall three dimensional arrangement can be deduced from the X-ray and is in accordance with previous work based upon bacteriorhodopsin receptor which is not G-protein-coupled.
GPCRs are most often characterised by sequence homology as being comprised of several sub-families. Most attention currently is directed towards Family A receptors as being the most tractable class historically and also the one with the most potential targets .
Family A comprises about 300 receptors that are potential drug targets, approximately half of which have known ligands and the rest, the so-called orphan receptors. The group of druggable receptors is composed essentially of two types: those whose natural ligand interacts wholly within the transmembrane domain, such as the aminergic, nucleotide-like, prostaglandin receptors, etc. and those peptide liganded receptors, which have a large part of their interactions in the extracellular region and which may insert a peptide loop or tail into the transmembrane region to effect signal transduction. Examples of this class are angiotensin, cholecystokinin and opioid receptors. Irrespective of the mode of action of the natural ligand or the GPCR family, the vast majority of drug molecules interact in the all-helical domain of the transmembrane region with exceptions being those mimics of glutamate at the metabotropic glutamate receptor and some peptide therapeutics administered parenterally. In looking for lead molecules for an unexplored or orphan GPCR it therefore makes sense to concentrate on interactions in the transmembrane domain.
The focused library provided herein is designed to interact with a range of the family A receptors . Each library is a defined set of compounds which will enhance the probability of finding a small molecule which will interact with one or more type of GPCR receptor.
For example, focused libraries can be provided having compounds which will interact with aminergic GPCRs, and peptidic GPCRs requiring an obligatory positive charge in ligands, or other types or groups of GPCRs .
Focused libraries according to this invention can provide hit rates of 1-13% or more for the requisite predicted GPCRs from both amine- and peptide-liganded classes and with agonists and antagonists .
Summary of Invention
We provide herein a "focused" library of compounds which will provide "leads" for ligands which bind to Family A G-Protein coupled receptors .
In the context of the present invention, "library" means a group of compounds which are structurally related by virtue of a core chemical structure (or "scaffold") but which differ from each other by virtue of permutation of specific substituent groups attached to the scaffold.
Generally speaking such a library will consist of or comprise a number of compounds, e.g. as many as about 100, 1000,2000, 3000 or indeed 10,000 compounds. The number of compounds should be sufficient to provide an adequate diversity of related compounds without being so large as to be unduly complex/expensive to produce .
In the context of the present invention the terms "permitted substituents" and analogous terms are used to refer to defined chemical groups which may be attached to a "scaffold" to provide permutations of the chemical structure of related compounds .
Where the chemical formulae of permitted substituents are shown in this description and claims, the substituent may appear in the compound exactly as shown (i.e. simply covalently bonded to the scaffold) or may be a derivative of the shown chemical formula of the substituent by virtue of use of a reactive group to couple the substituent to the scaffold.
It will be appreciated that the total number of permutations created by the permitted substituents may be a very large number, far greater in magnitude than the actual number of compounds in an actual library. In other words, the number of possible compounds for any "virtual" library may well greatly exceed the number of synthesised compounds making up an embodiment of the "real" library. The invention is intended to encompass libraries having all, and a number, which is less than all, of the permitted substitutions represented by compounds therein.
It will be appreciated that some specific combinations of permitted substituents may be more or less difficult to synthesise and/or use in a focused library of the invention. This does not detract from the generality of applicability of the invention as described herein. It is to be expected that real libraries will be synthesised from a selected group of permutations/combinations of permitted substituents, taking into consideration factors affecting the intended purpose of the library and its cost and complexity of synthesis. Even if theoretically permitted, it is currently considered unlikely that any compound would be prepared for inclusion in a focused library if it had either or both of the following properties
(1) molecular weight >700
(2) log p <-3 or >9 (an index of lipophilicity as calculated using commercially available "Chemenlighten 2.8" and "Biobyte" software for the log p calculation) .
The present invention provides novel focused libraries of compounds . Most of the compounds defined by the permitted substitutions on the scaffolds are also novel compounds per se and the invention is intended to encompass each individual novel compound. Any known compound having a structural formula identical to any one of the compounds covered by the formulae of scaffolds and permitted substitutions described herein is hereby explicitly disclaimed per se.
DESCRIPTION OF THE INVENTION
Library 7 (SFG07) is designed to target a large group of receptors that recognise electron rich aromatic ring systems. These receptors fall into two main groups, those that also require a positively charged amine in the ligand (for example the adrenoceptors or the serotonin receptors) and those that do not (for example the melanocortin, vasopressin, and melatonin receptors) .
The invention provides a compound comprising or consisting of a set of structurally related compounds of general formula (I) :
Figure imgf000008_0001
(I)
wherein the permitted substituents for Rl are derived from the precursor groups of List 1 and may be further derivatised by reaction with the boronic acids of List 6, the permitted substituents for R2 are derived from the precursor groups of Lists 2-5, and n = 0-4.
Structural Novelty of Compounds of Library 7
Figure imgf000008_0002
The following compounds are known in the literature, these compounds are not permitted within Library SFG07:
Figure imgf000009_0001
Figure imgf000009_0002
The following three structures are known in the literature, these structures could be contained in the SFG07 library but they are disclaimed per se.
Figure imgf000009_0003
Methods for synthesising Scaffolds of Library 7 Compounds of formula I can be made according to the following general scheme (Scheme 1)
Figure imgf000010_0001
(il) (III)
Figure imgf000010_0002
Acylation Alkylation " Reaction with isocyanates
Figure imgf000010_0003
( Scheme 1)
The 4H-spiro [1, 3-benzodioxine-2, 4 ' -piperidine] (IV) can be prepared from an appropriately substituted 2-hydroxybenzyl alcohol (II) , selected from List 1, by reaction with a suitably N-protected 4-piperidinone (III) followed by appropriate deprotection. Suitable protecting groups would include ethoxy carbonyl, benzoxy carbonyl, acetyl, benzyl. The spirol,3- benzodioxane piperidines (IV) can then be further derivatised by reaction at the secondary nitrogen which can include alkylation by reaction with a compound selected from List 2 (R2-# where # is a leaving group especially halogen or sulphonate) to give (la) (R2 = optionally substituted alkyl) , acylation by reaction with a compound selected from List 3 (R2-C1) to give (lb) (R2 = Acyl) , reaction with an isocyanate selected from List 4 (X-NCO) to give (Ic) (R2 is XNHC(=0)-) or reaction with a cyclic anhydride selected from List 5 to give (Id) (R2= -C (=0) -Y-C (=0) OH) . The 4H- spiro [1,3 -benzodioxine-2, 4 ' -piperidine] (IV) can be further derivatised by modification of the Rl group. The Rl group may be modified by a number of reactions, including so called Suzuki reactions and other palladium catalysed reactions.
Figure imgf000012_0001
Figure imgf000012_0002
Figure imgf000012_0003
Figure imgf000012_0004
When Rl is a bromine atom, 4H-spiro [1, 3-benzodioxine-2 , 4 ' - piperidine] (IV) may be reacted with a variety of boronic acids selected from List 6 to give compounds where the bromine is replaced by an aryl or heteroaryl group for example see Scheme 2.
Figure imgf000013_0001
Scheme 2
Other methods for the synthesis of the intermediates will be apparent to the chemist skilled in the art, as will be the methods for preparing starting materials and intermediates . The isolated novel compounds were confirmed by 1H N.M.R. and/or other appropriate methods .
In the priority application (GB 0313762.7) where hydroxyl or halogen groups are shown in places of #s, the hydroxyl and halogen groups may be modified to form leaving groups.
The permitted substituents at positions Rl and R2 are for compounds of Library 7 (SFG07) are shown below.
Figure imgf000014_0001
Figure imgf000014_0002
Figure imgf000014_0003
List 1
Figure imgf000015_0001
List 2 (R2-#)
Figure imgf000016_0001
List 2 (cont'd)
Figure imgf000017_0001
List 2 (cont'd) 17
Figure imgf000018_0001
Figure imgf000019_0001
List 3 (cont'd)
Figure imgf000020_0001
List 3 (cont'd)
Figure imgf000021_0001
Figure imgf000021_0003
Figure imgf000021_0004
Figure imgf000021_0002
List 3 (cont'd)
Figure imgf000022_0002
Figure imgf000022_0003
Figure imgf000022_0001
List 3 (cont'd)
Figure imgf000023_0001
List 3 (cont'd)
Figure imgf000024_0001
Figure imgf000024_0002
Figure imgf000024_0003
Figure imgf000024_0004
Figure imgf000025_0001
List 3 (cont'd)
Figure imgf000026_0002
Figure imgf000026_0001
Figure imgf000026_0003
Figure imgf000026_0004
List 3 (cont'd)
Figure imgf000027_0001
Figure imgf000027_0002
Figure imgf000027_0003
List 3 (cont'd)
Figure imgf000028_0001
Figure imgf000028_0002
Figure imgf000028_0003
List 4 (X-NCO)
Figure imgf000029_0001
Figure imgf000029_0002
Figure imgf000029_0003
Figure imgf000029_0004
List 4 (cont'd)
Figure imgf000030_0001
Figure imgf000030_0002
Figure imgf000030_0003
Figure imgf000030_0004
List 4 (cont'd)
Figure imgf000031_0001
Figure imgf000031_0002
List 4 (cont'd)
Figure imgf000032_0001
Figure imgf000032_0002
Figure imgf000033_0001
Figure imgf000033_0002
Figure imgf000033_0003
H
0
Figure imgf000033_0004
Figure imgf000033_0005
Figure imgf000033_0006
List 6 (Rl-B(OH)2)
Figure imgf000034_0001
Figure imgf000034_0002
Figure imgf000034_0003
List 6 (cont'd)
Figure imgf000035_0001
Figure imgf000035_0002
Figure imgf000035_0003
List 6 (cont'd)
Figure imgf000036_0001
List 6 (cont'd)
Figure imgf000037_0001
Figure imgf000037_0002
List 6 (cont'd)
Figure imgf000038_0001
Figure imgf000038_0002
List 6 (cont'd)
Figure imgf000039_0001
Figure imgf000039_0002
List 6 (cont'd)
Figure imgf000040_0001
Figure imgf000040_0002
List 6 (cont'd)
Specific Examples of Compounds of Library 7
Synthesis of 6-fluoro-4H-spiro [1, 3-benzodioxine-2,4' -piperidine]
Figure imgf000041_0001
To a solution of 5-fluoro-2-hydroxybenzyl alcohol (4.0g, 28mmol) and N-carboethoxy-4-piperidone (6.3g, 37mmol) in chloroform
(60ml) was added p-toluenesulphonic acid (0.5g). The mixture was heated at reflux for 18hrs using a Dean-Stark condenser to collect the water formed by the reaction. The • solvent was removed in vacuo . The residue was dissolved in ethyl acetate
(150ml) and washed with 2M sodium hydroxide solution (100 ml) and brine (100ml) . The organic layer was dried with magnesium sulfate and the solvent removed in Vacuo. The residue was purified by column chromatography using a mixture of hexane and ethyl acetate to give (4.86g; 58%) of 1' -carboethoxy-6-fluoro-4iϊ- spiro [1, 3-benzodioxine-2 , 4' -piperidine] . To a solution of this product in ethanol (100ml) was added 6M sodium hydroxide solution
(35ml) and the mixture was heated at reflux for 20hrs. The mixture was evaporated in vacuo and the residue was partitioned between ethyl acetate (100 ml) and water (100 ml) . The aqueous layer was extracted with ethyl acetate (100ml) and the combined organic layer was washed with brine (100ml) and dried with magnesium sulfate . The solvent was removed in vacuo to give
(3.37g; 54 % overall yield) of the title compound; HPLC (88% Rτ *= 1.63min) ; XH NMR (CDC13) : δ= 1.23-1.32 (m, 4H, 2xCH2) , 2.94- 3.04 (m, 4H, 2xCH2) , 4.82 (s, 2H, CH20) , 6.66-6.71 (m, 1H, ArH) , 6.78-6.92 (m, 2H, ArH); "C NMR (CDC13) : δ= 34.5, 42.9, 59.9, 98.2, 110.7, 111.1, 114.7, 115.1, 118.2, 120.7, 146.6, 154.9, 158.7; MS (electrospray; [M+H]+) m/z: 224. Synthesis of 8-methoxy-4H-spiro [1, 3-benzodioxine-2,4' -piperidine]
Figure imgf000042_0001
To a solution of 2-hydroxy-3-methoxybenzyl alcohol (10. Og, 65mmol) and N-carboethoxy-4-piperidone (12. Og, 70mmol) in chloroform (130ml) was added p-toluenesulphonic acid (0.5g). The mixture was heated at reflux for 18hrs using a Dean-Stark condenser to collect the water formed by the 'reaction. The solvent was removed in vacuo . The residue was dissolved in ethyl acetate (200ml) and washed with 2M sodium hydroxide solution (100 ml) and brine (100ml) . The organic layer was dried with magnesium sulfate and the solvent removed in vacuo . The residue was purified by column chromatography using a mixture of hexane and ethyl acetate to give (8.70g; 44%) of 1' -carboethoxy-8-methoxy- 4-ϊ-spiro [1, 3 -benzodioxine-2, 4' -piperidine] . To a solution of this product in ethanol (200ml) was added 4M sodium hydroxide solution
(60ml) and the mixture was heated at reflux for 48hrs. The solvent was removed in vacuo and the residue was partitioned between ethyl acetate (200 ml) and water (100 ml) . The aqueous layer was extracted with ethyl acetate (100ml) and the combined organic layer was washed with brine (100ml) and dried with magnesium sulfate. The solvent was removed in vacuo to give
(6.35g; 42 % overall yield) of the title compound; HPLC (98% Rτ = 1.39min); E NMR (CDC13) : δ= 1.81-1.89 (m, 4H, 2xCH2) , 2.82- 3.00 (m, 4H, 2 CH2) , 3.80 (s, 3H, 0CH3) , 4.77 (s, 2H, CH20) , 6.49- 6.52 (m, 1H, ArH), 6.68-6.82 (m, 2H, ArH); 13C NMR (CDC13) : δ= 43.1, 56.0, 59.9, 98.5, 110.2, 116.4, 120.1, 120.6, 140.3, 148.6; MS (electrospray; [M+H]+) m/z: 236. Synthesis of 6-methoxy-4H-spiro [1, 3-benzodioxine-2, ' -piperidine]
Figure imgf000043_0001
To a solution of 2-hydroxy-5-methoxybenzyl alcohol (10. Og, 65mmol) and N-carboethoxy-4-piperidone (13.3g, 78mmol) in chloroform (125ml) was added p-toluenesulphonic acid (0.62g). The mixture was stirred at room temperature for 18hrs. The solvent was removed in vacuo. The residue was dissolved in ethyl acetate (200ml) and washed with 2M sodium hydroxide solution (125 ml) . The organic layer was dried with magnesium sulfate and the solvent removed in vacuo. The residue was purified by column chromatography using a mixture of hexane and ethyl acetate to give (5.12g; 26%) of 1' -carboethoxy-6-methoxy-4H-spiro [1, 3- benzodioxine-2, 4' -piperidine] . To a solution of this product in. ethanol (160ml) was added 3.3M sodium hydroxide solution (40ml) and the mixture was heated at reflux for 48hrs. The mixture was evaporated in vacuo and the residue was partitioned between ethyl acetate (200 ml) and water (100 ml) . The organic layer was dried with magnesium sulfate and the solvent was removed in vacuo to give (2.70g; 18% overall yield) of the title compound; HPLC (88%; Rτ = 1.55min); lH NMR (CDC13) : δ= 1.23-1.30 (m, 4H, 2xCH2) , 2.91- 3.01 ( , 4H, 2xCH2), 3.75 (S, 3H, 0CH3) , 4.83 (s, 2H, CH20) , 6.51 (d, 1H, J= 2.6 Hz, ArH), 6.72-6.83 (m, 1H, ArH); 13C NMR (CDC13) : δ= 34.5, 43.0, 55.7, 60.2, 60.4, 97.9, 109.2, 114.2, 117.8, 120.3, 144.5, 153.5; MS (electrospray; [M+H]+) m/z: 236.
Synthesis of 6-thien-2-yl-4H-spiro [l,3-benzodioxine-2,4' - piperidine]
Figure imgf000044_0001
To a solution of 6-bromo-4H-spiro [1, 3-benzodioxine-2 , 4 ' - piperidine] (5.68g, 20mmol) , 2-thiophene boronic acid (3.07g, 24mmol) , triphenyl phosphine (0.78g, 3mmol) and sodium hydrogencarbonate (6.4g, 60mmol) in ethanol/water (4:1) (100ml) was added palladium (II) acetate (0.22g, lmmol) . The mixture was heated at reflux under a nitrogen atmosphere for 18hrs. The mixture was evaporated in vacuo and the residue suspended in dichloromethane (100ml) filtered and the filtrate washed with 2M sodium hydrogencarbonate solution (80ml) and brine (80ml) . The organic layer was dried with magnesium sulfate and evaporated in vacuo and the crude material purified by column chromatography using a mixture of dichloromethane and methanol to give (1.84g, 33%) of the title compound as a yellow gum; HPLC (100%; Rτ =1.99min) ; **Η NMR (CDC13) : δ= 1.81-1.99 (m, 4H, 2xCH2) , 2.93-3.15 (m, 4H, 2xCH2) , 4.88 (s, 2H, CH20) , 6.49 (d, 1H, J 8.5 Hz, ArH), 7.02-7.06 (dd, 1H, J 5.05 & 3.62 Hz, ArH), 7.15-7.25 (m, 3H, ArH), 7.39-7.44 (dd, 1H, J 8.50 & 2.25 Hz, ArH); 13C NMR (CDC13) : δ= 34.5, 42.9, 60.0, 98.4, 117.6, 120.0, 122.2, 122.2, 123.9, 126.0, 127.1, 127.9, 144.1, 150.0; MS (electrospray; [M+H]+) m/z: 288.
Synthesis of 6- (2-phenoxyphenyl) -4H-spiro [1, 3-benzodioxine-2,4' - piperidine]
Figure imgf000045_0001
To a solution of 6-bromo-4H-spiro [1, 3-benzodioxine-2,4 ' - piperidine] (l.OOg, 3.5mmol), 2-phenoxyphenylboronic acid (0.90g, 4.2mmol), triphenyl phosphine (0.13g, 0.52mmol) and sodium hydrogencarbonate (1.06g, lOmmol) in ethanol/water (4:1) (80ml) was added palladium (II) acetate (0.22g, lmmol) . The mixture was heated at reflux under a nitrogen atmosphere for lδhrs. The mixture was evaporated in vacuo and the residue suspended in dichloromethane (80ml) filtered and the filtrate washed with 2M sodium hydrogencarbonate solution (60ml) and brine (60ml) . The organic layer was dried with magnesium sulfate and the solvent removed in vacuo and the crude material purified by column chromatography using a mixture of dichloromethane and methanol to give (0.48g, 36.5%) of the title compound as an orange gum; HPLC
(99.24%; Rτ 2.40min); *Η NMR (CDCl3 δ= 1.85-2.01 (m, 4H, 2xCH2) ,
2.98-3.02 (m, 4H, 2xCH2) , 3.39 (br s, 1H, NH) , 4.82 (s, 2H, CH20), 6.82-7.38 (m, 12H, ArH); *L3C NMR (CDCl3) : δ= 34.0, 42.6, 60.17, 67.06, 97.87, 116.8, 118.0, 199.1, 120.1, 122.6, 124.0, 125.4, 128.3, 129.1, 129.5, 130.1, 130.9, 133.0, 149.9, 153.4, 157.7; MS (electrospray; [M + H]+) m/z: 374.
Synthesis of 1' -benzyl-6- (3 , 4-dimethylphenyl) -4H-spiro [1, 3- benzodioxine-2 , 4 ' -piperidine]
Figure imgf000046_0001
To a solution of benzyl bromide (0.037g, 0.22mmol) in iso- propanol (0.5ml) was added 6- (3 , 4-dimethylphenyl) -4H-spiro [1, 3- benzodioxine-2, 4' -piperidine] (0.062g, 0.2mmol) in 0.6M solution of N, N,N-di±sopropylethyl amine in iso-propanol (0.5ml). The mixture was heated at 80°C for 18hrs . The solvent was removed in vacuo. The crude product was purified by preparative HPLC (see Appendix 1); HPLC .(98%, Rt= 2.79 min) ; E NMR (CDC13) δ= 1.99-2.06 (m, 4H, 2xCH2) , 2,30 (d, 6H, J= 6.0Hz, 2xCH3 , ) , 2.56-2.58 (m, 4H, 2xCH2) , 3.57 (s, 2H, CH2) , 4.89 (s, 2H, 0CH2) , 6.92 (d, 1H, J=8.5Hz, ArH), 7.12-7.16 (m, 2H, ArH), 7.21-7.26 (m, 3H, ArH), 7.29-7.36 (m, 5H, ArH); MS (electrospray [(M+H)+]) m/z:400.
Synthesis of 1' - [2- (Uϊ-indol-3-yl) ethyl] -6- (2-phenoxyphenyl) -4H- spiro [1,3 -benzodioxine-2,4' -piperidine]
Figure imgf000046_0002
To a solution of 3- (2-bromoethyl) indole (0.049g, 0.22mmol) in iso-propanol (0.5ml) was added 6- (2-phenoxyphenyl) -4H-spiro [1,3- benzodioxine-2, 4' -piperidine] (0.074g, 0.2mmol) in 0.6M solution of N, N, -diisopropylethyl amine in iso-propanol (0.5ml). The mixture was heated at 80°C for 18hrs . The solvent was removed in vacuo . The crude product was purified by preparative HPLC (see Appendix 1); Yield = 0.051g (49.5%); HPLC (88.1%; Rτ 4.79min) ; E NMR (CDC13) : δ= 1.95-2.08 (m, 4H, 2xCH2) , 2.61-2.74 (m, 4H, 2xCH2) , 2.79 (d, 2H, J 9.4 Hz, CH2) , 2.99 (d, 2H, J 9.2 Hz, CH2) , 4.84 (s, 2H, 0CH2) , 6-75-7.41 (m, 16H, ArH), 7.61 (d, 1H, J 7.61 Hz, ArH), 8.07 (br s, 1H) ; 13C NMR (CDC13) : δ 23.2, 33.3, 49.8, 58.7, 60.3, 67.0, 98.1, 111.1, 114.3, 116.8, 118.1, 118.8, 119.2, 119.3, 120.1, 121.4, 121.9, 122.6, 124.0, 125.4, 127.4, 128.3, 129.0, 129.5, 130.0, 131.0, 133.0, 136.2, 150.1, 153.4, 157.7; MS (electrospray; [M+H]+) m/z:517.
Synthesis of 2- (6-bromo-l'iϊ, 4iϊ-spiro [1/3 -benzodioxine-2, 4 ' piperidin] -1' -yl) -1- (4 -fluorophenyl) ethanone
Figure imgf000047_0001
To a solution of 4-fluorophenylacyl bromide (0.048g, 0.22mmol) in iso-propanol (0.5ml) was added 6-bromo-4H-spiro [1, 3-benzodioxine- 2,4' -piperidine] (0.056g, 0.2mmol) in a 0.6M solution of N, N, N- diisopropylethyl amine in iso-propanol (0.5ml). The mixture was heated at 80°C for 18hrs. The solvent was removed in vacuo . The crude product was purified by preparative HPLC (see Appendix 1) ; Yield (0.051g, 59%); HPLC (98.2%, Rt= 2.12min) ; E NMR (CDCl3) δ= 1.92-2.02 (m, 4H, 2xCH2) , 2.60-2.69 (m, 4H, 2xCH2) , 3.83 (s, 2H, CH2) , 4.81 (s, 2H, CH20) , 6.78 (d, 1H, J= 8.7 Hz, ArH), 7.10 (s, 1H, ArH), 7.13-7.19 (m, 2H, ArH), 7.24-7.27 (m, 1H, ArH), 8.03- 8.11 (m, 2H, ArH); MS (electrospray, [M+H] + ) m/z:420/422. Synthesis of 1" -benzoyl-6- [4- (trifluoromethoxy)phenyl] -4H- spiro [1, 3 -benzodioxine-2 , 4 ' -piperidine]
Figure imgf000048_0001
To a solution of benzoyl chloride (0.031g, 0.22mmol) in tetrahydrofuran (0.5ml) was added a solution of 6- [4- (trifluoromethoxy)phenyl] -4H-spiro [1, 3 -benzodioxine-2, ' - piperidine] (0.073g, 0.2mmol) in a 0.6M solution ^ of N, N, N- diisopropylethyl amine in tetrahydrofuran (0.5ml). The mixture was agitated for 16hrs under an atmosphere of nitrogen. The solvent was removed in vacuo and the product purified by preparative HPLC (see Appendix 1 for conditions) ; 1H NMR (CDC13) δ= 1.93 (br s, 4H, 2xCH2) , 2.05 (br s, 4H, 2xCH2) , 4.93 (d, 2H, J= 3.7 Hz, CH20) , 6.95 (d, 1H, J= 8.5 Hz, ArH), 7.16 (d, 1H, J= 1.6 Hz, ArH), 7.25 (d, 2H, J= 8.1 Hz, ArH), 7.36-7.43 (m, 6H, ArH), 7.51 (d, 2H, J= 8.7 Hz, ArH); 13C NMR (CDC13) δ= 60.4, 98.1, 117.6, 119.8, 121.2, 123.4, 126.8, 127.1, 128.0, 128.5, 129.8, 132.8, 135.8, 139.2, 148.3, 150.3, 170.5; MS (electrospray, [M+H]+) m/z: 470. Synthesis of 6-phenyl-l' - (thien-2-ylcarbonyl) -4H-spiro [1, 3- benzodioxine-2 , 4 ' -piperidine]
Figure imgf000049_0001
To a solution of thiophene-2-carbonyl chloride (0.032g, 0.22mmol) in tetrahydrofuran (0.5ml) was added a solution of 6-phenyl-4H- spiro [1, 3 -benzodioxine-2 , 4 ' -piperidine] (0.056g, 0.2mmol) in a 0.6M solution of JNT/N,.N-diisopropylethyl amine in tetrahydrofuran (0.5ml). The mixture was agitated for 16hrs under an atmosphere of nitrogen. The solvent was removed in vacuo and the product purified by preparative HPLC (see Appendix 1 for conditions) ; Ή
NMR (CDC13) δ= 1.94-2.17 (m, 4H, 2xCH2) , 3.71-3.84 (m, 2H, CH2) , 3.91-4.01 (m, 2H, CH2) , 4.95 (s, 2H, CH20) , 6.95 (d, 1H, J= 8.5 Hz, ArH), 7.06 (dd, 1H, J= 5.0 & 3.7 Hz, ArH), 7.21 (d, 1H, J= 2.1 Hz, ArH), 7.27-7.54 (m, 8H, ArH); 13C NMR (CDC13) δ= 60.5, 98.0, 117.5, 119.6, 123.4, 126.7, 126.9, 127.1, 128.6, 128.8, 134.2, 137, 140.5, 149.9, 163.8; MS (electrospray, [M+H] +) m/z: 392.
Synthesis of 1' - (3 -methyl-benzoyl) -6- [4- (trifluoromethoxy) henyl] -4H-spiro [1, 3 -benzodioxine-2, 4 ' -piperidine] .
Figure imgf000049_0002
To a solution of 3 -methyl-benzoyl chloride (0.034g, 0.22mmol) in tetrahydrofuran (0.5ml) was added a solution of 6-(3,4- dimethoxy)phenyl-4H-spiro [1, 3 -benzodioxine-2 , 4 ' -piperidine] (0.068g, 0.2mmol) in a 0.6M solution of N, N, N-diisopropylethyl amine in tetrahydrofuran (0.5ml). The mixture was agitated for 16hrs under an atmosphere of nitrogen. The solvent was removed in vacuo and the product purified by preparative HPLC (see Appendix 1 for conditions); XH NMR (CDCl3) δ= 1.92-2.17 (m, 4H, 2xCH2) , 2.38 (s, 3H, CH3) , 3.50-3.90 (m, 4H, 2xCH2) , 3.91 (s, 3H, CH30) , 3.94 (s, 3H, CH30) , 4.93 (d, 2H, J= 4.6 Hz, CH20) , 6.90-7.42 (m, 10H, ArH); 13C NMR (CDCl3) δ= 21.3, 55.9, 56.0, 60.5, 98.0, 110.1, 111.5, 117.4, 118.9, 119.6, 123.0, 123.7, 126.9, 127.4, 128.3, 130.4, 133.6, 134.1, 135.8, 138.4, 148.3, 149.1. 149.6, 170.7; MS (electrospray, [M+H]+) m/z: 460.
Synthesis of N- (4-methoxyphenyl) -1 'iϊ,4iT-spiro [1, 3-benzodioxine- 2,4 ' -piperidine] -1' -carboxamide
Figure imgf000050_0001
To a solution of l-isocyanato-4-methoxy-benzene (0.033g, 0.22mmol) in tetrahydrofuran (0.5ml) was added a solution of 4Jϊ- spiro [1, 3 -benzodioxine-2 , 4 ' -piperidine] (0.041g, 0.2mmol) in tetrahydrofuran (0.5ml). The mixture was agitated for 16hrs under an atmosphere of nitrogen. The solvent was removed in vacuo and the product purified by preparative HPLC (see Appendix 1 for conditions); Yield (0.041g, 58%); HPLC (88.3%, Rt= 3.47min); 1H NMR (CDCI3) 5=1.74-1.88 (4H, m) , 3.39-3.60 (4H, m) , 3.68 (3H, s) , 4.85 (2H, s), 6.77-7.34 (8H, m) , 8.45 (1H, s) ; 13C NMR (CDC13) -. δ= 32.9, 40.6, 55.0, 59.5, 66.3, 97.9, 113.4, 116.5, 119.9, 120.5, 121.6, 125.1, 127.9, 133.4, 150.1, 154.4, 155.1; MS (electrospray, [M+H]+) m/z: 355.
Synthesis of iV- (3-chlorophenyl) -l'ff, 4iT-spiro [1, 3-benzodioxine- 2,4' -piperidine] -1' -carboxamide
Figure imgf000051_0001
To a solution of l-isocyanato-3-chloro-benzene (0.033g, 0.22mmol) in tetrahydrofuran (0.5ml) was added a solution of 4H-spiro [1, 3- benzodioxine-2 , 4 ' -piperidine] (0.041g, 0.2mmol) in tetrahydrofuran (0.5ml). The mixture was agitated for 16hrs under an atmosphere of nitrogen. The solvent was removed in vacuo and the product purified by preparative HPLC (see Appendix 1 for conditions); Yield (0.041g, 58%); HPLC (85.2%, Rt= 3.87min); 1H NMR (CDCI3) 5=1.77-1.86 (m, 4H, 2xCH3) , 3.50-3.57 (m, 4H, 2xCH3) , 4.86 (s, 2H, CH20) , 6.81-7.39 (m, 7H, ArH), 7.63-7.64 (m, 1H, ArH), 8.80 (s, 1H, ArH); 13C NMR (CDCl3) : δ = 32.9, 40.6, 59.5, 66.3, 97.8, 116.5, 117.6, 118.7, 119.8, 120.5, 121.2, 125.1, 128.1, 129.8, 132.6, 142.1, 150.1, 154.4; MS (electrospray, [M+H]+) m/z: 359/361.
Synthesis of N- (3 , 5-dimethylisoxazol-4-yl) -l'JT, 4_ϊ-spiro [1, 3- benzodioxine-2, 4 ' -piperidine] -1' -carboxamide
Figure imgf000052_0001
To a solution of 4-isocyanato-3 , 5-dimethyl-isoxazole (0.030g, 0.22mmol) in tetrahydrofuran (0.5ml) was added a solution of 4H- spiro [1, 3 -benzodioxine-2, 4 ' -piperidine] (0.041g, 0.2mmol) in tetrahydrofuran (0.5ml). The mixture was agitated for 16hrs under an atmosphere of nitrogen. The solvent was removed in vacuo and the product purified by preparative HPLC (see Appendix 1 for conditions); Yield (0.041g, 58%); HPLC (99.9%, Rt= 3.05min); 1H NMR (CDC13) 5=1.81-1.84 (m, 4H, 2xCH2) , 2.04 (s, 3H, CH3) , 2.11 (s, 3H, CH3) , 3.48-3.53 (m, 4H, 2xCH2) , 4.86 (s, 2H, CH20) , 6.82- 7.19 (m, 4H, ArH), 8.02 (s, 1H, ArH); 13C NMR (CDC13) : 5= 9.4, 10.5, 32.9, 40.7, 59.5, 97.8, 115.6, 116.5, 119.8, 120.5, 125.1, 128.0, 150.1, 155.4, 158.3, 162.1; MS (electrospray, [M+H]+) m/z: 344.
Synthesis of 2- [ (6-fluoro-1'.ff, 42ϊ-spiro [1, 3 -benzodioxine-2, 4 ' - piperidin] -1 ' -yl) carbonyl] benzoic acid
Figure imgf000052_0002
To a solution of isobenzofuran-1, 3-dione (0.035g, 0.22mmol) in dimethylformamide (0.5ml) was added a solution of 6-fluoro-4H- spiro [1, 3 -benzodioxine-2, 4 ' -piperidine] (0.045g, 0.2mmol) in dimethylformamide , (0.5ml). The mixture was agitated at room temperature for 18hrs . Product was purified by preparative HPLC (see Appendix 1 for conditions); Yield (0.040g, 54%); 1H NMR (CDC13) 5= 1.66-2.17 (m, 4H, 2xCH2) , 3.60-3.90 (m, 4H, 2xCH2) , 4.67 (br s, 2H, CH20) , 6.48-6.84 (m, 3H, ArH), 7.18 (d, 1H, J= 7.3 Hz, ArH), 7.30-7.36 (m, 1H, ArH), 7.41-7.47 (m, 1H, ArH), 7.98 (d, 1H, J= 7.4 Hz, ArH); 13C NMR (CDC13) : 5= 11.6, 18.2 32.0, 33.1, 38.2, 41.2, 43.7, 52.6, 60.0, 98.1, 111.1, 114.7, 118.1, 125.8, 131.1, 132.2, 137.6, 146.3, 170.3, 171.2; MS (electrospray, [M+H]+) m/z: 372.
Synthesis of [2- (6-methoxy-1'IT, 4H-spiro [1, 3 -benzodioxine-2, 4 ' - piperidin] -1' -yl) -2 -oxoethoxy] acetic acid
Figure imgf000053_0001
To a solution of [1, 4] dioxane-2, 6-dione (0.026g, 0.22mmol) in dimethylformamide (0.5ml) was added a solution of 6-methoxy-4H- spiro [1, 3 -benzodioxine-2 , 4 ' -piperidine] (0.047g, 0.2mmol) in dimethylformamide (0.5ml). The mixture was agitated at room temperature for 18hrs. Product was purified by preparative HPLC (see Appendix 1 for conditions); Yield (0.041g, 59%); XH NMR (CDC13) 5= 1.86 (br s, 4H, 2xCH2) , 3.48 (br s, 4H, 2xCH2) , 3.73 (s, 3H, CH3O) , 3.96 (s, 2H, CH20) , 4.26 (s, 2H, CH20) , 4.76 (s, 2H, CH20) , 6.48 (d, 1H, J= 2.5 Hz, ArH), 6.68-6.74 (m, 2H, ArH); 13C NMR (CDCI3) : 5= 18.2, 30.3, 32.5, 33.8, 38.5, 41.0, 52.4, 55.7, 60.4, 68.9, 70.5, 97.4, 109.2, 114.4, 117.8, 120.0, 144.1, 153.7, 168.3, 175.2; MS (electrospray, [M+H] +) m/z: 352.
Synthesis of {1- [2- (8-methyl-6- ( ethylsulfanyl) -42ϊ-spiro [1,3- benzodioxine-2 , 4 ' -piperidine] ) -2 -oxoethyl] cyclopentyl}acetic acid
Figure imgf000054_0001
To a solution of 8-oxa-spiro [4.5] decane-7, 9-dione (0.037g, 0.22mmol) in dimethylformamide (0.5ml) was added a solution of 8- methyl-6- (methylsulfanyl) -4---.-spiro [1, 3 -benzodioxine-2 ,4 ' - piperidine] (0.053g, 0.2mmol) in dimethylformamide (0.5ml) . The mixture was agitated at room temperature for lδhrs. Product was purified by preparative HPLC (see Appendix 1 for conditions) ; Yield (0.053g, 61%); 1H NMR (CDC13) 5= 1.45-2.15 (m, 10H, 5xCH2) , 2.19 (s, 3H, CH3) , 2.44 (s, 3H, CH3S) , 2.49 (s, 2H, CH2) , 2.62 (s, 2H, CH2) , 3.48 to 3.67 (m, 6H, 3xCH2) , 4.06-4.15 (m, 1H) , 3.76- 3.84 (m, 1H) , 6.79 (d, 1H, J= 1.7 Hz, ArH), 7.02 (s, 1H, ArH); 13C NMR (CDC13) : 5= 15.3, 17.6, 23.8, 32.8, 34.6, 38*.5, 38.6, 38.7, 39.1, 43.8, 44.3, 45.0, 60.3, 97.3, 119.4, 122.1, 127.1, 129.1, 129.7, 146.6, 172.0, 173.0; MS (electrospray, [M+H]+) m/z 434.
Appendix 1
Analytical HPLC Conditions
Figure imgf000055_0001
Table 1
Prep HPLC Conditions
All compounds were purified using reverse phase HPLC using a Gilson preparative HPLC system (322 pump, 155 UV/VIS detector, 215 liquid handler) and a Xterra MS, 100 x 19 mm, C18, 5 μm column. A flow rate of 25 mL/min was used.
The Gilson 215 was used as both auto-sampler and fraction collector
The gradient used was 90% water (5 mM ΝH4HC03) / 10% methanol for 1.5 min to 100% methanol over 6.5 min then held at 100% methanol for 3.0 min. The solvent mixture was then returned to the initial conditions over 0.5 min.
The purification was controlled by Unipoint software, triggering a threshold collection value monitoring at 220 nm. Collected fractions were analysed by LCMS (Waters/Micromass ZQ) (Table 1) . The fractions that contained the desired product are concentrated by vacuum centrifugation and the resultant residue dried by freeze-drying .

Claims

A compound library comprising' or consisting of a set of structurally related compounds having a core chemical structure (scaffold) of general formula (I) •
Figure imgf000057_0001
(I) wherein the permitted substituents for Rl are derived from the precursor groups of List 1 and may be further derivatised by reaction with the boronic acids of List
6; the permitted substituents for R2 are derived from the precursor groups of Lists 2-5, # is a leaving group; and n = 0-4.
Figure imgf000058_0001
Figure imgf000058_0002
Figure imgf000058_0003
Figure imgf000058_0004
Figure imgf000058_0005
Figure imgf000058_0006
List 1
Figure imgf000059_0001
List 2 (R2-#)
Figure imgf000060_0001
List 2 (cont'd)
Figure imgf000061_0001
List 2 (cont'd)
Figure imgf000062_0001
List 3 (R2-C1)
Figure imgf000063_0001
Figure imgf000063_0003
Figure imgf000063_0002
Figure imgf000063_0004
Figure imgf000063_0005
Figure imgf000064_0001
Figure imgf000065_0001
Figure imgf000065_0003
Figure imgf000065_0004
Figure imgf000065_0002
List 3 (cont'd)
Figure imgf000066_0002
Figure imgf000066_0003
Figure imgf000066_0001
List 3 (cont'd)
Figure imgf000067_0001
List 3 (cont'd)
Figure imgf000068_0001
Figure imgf000068_0002
Figure imgf000068_0003
Figure imgf000068_0004
List 3 (cont'd)
Figure imgf000069_0001
List 3 (cont'd)
Figure imgf000070_0001
List 3 (cont'd)
Figure imgf000071_0001
Figure imgf000071_0002
Figure imgf000071_0003
List 3 (cont'd)
Figure imgf000072_0001
Figure imgf000072_0002
O
//
List 4 (X-NCO)
Figure imgf000073_0001
Figure imgf000073_0002
Figure imgf000073_0003
Figure imgf000073_0004
List 4 (cont'd)
Figure imgf000074_0001
Figure imgf000074_0002
Figure imgf000074_0003
Figure imgf000074_0004
List 4 (cont'd)
Figure imgf000075_0001
Figure imgf000075_0002
List 4 (cont'd)
Figure imgf000076_0001
List 5
Figure imgf000077_0001
Figure imgf000077_0002
Figure imgf000077_0003
Figure imgf000077_0004
Figure imgf000077_0005
List 6 (Rl-B(OH)2)
Figure imgf000078_0001
Figure imgf000078_0002
List 6 (cont'd)
Figure imgf000079_0001
Figure imgf000079_0002
Figure imgf000079_0003
List 6 (cont'd)
Figure imgf000080_0001
List 6 (cont'd)
Figure imgf000081_0001
Figure imgf000081_0002
List 6 (cont'd)
Figure imgf000082_0001
Figure imgf000082_0002
List 6 (cont'd)
Figure imgf000083_0001
Figure imgf000083_0002
Figure imgf000083_0003
Figure imgf000083_0004
Figure imgf000083_0005
Figure imgf000083_0006
List 6 (cont'd)
Figure imgf000084_0001
Figure imgf000084_0002
List 6 (cont'd)
2. A library according to the claim 1 wherein said library has all or substantially all of the compounds represented therein.
3. A library according to claim 1 or 2 wherein said library has about 100, 1000, 2000, 3000, or 10000 of the compounds represented therein.
4. A method for making a compound library according to claim 1, which method comprises the step of synthesising compounds of formula I according to the following reaction scheme:
Figure imgf000086_0001
Figure imgf000086_0002
Acylation Alkylation ' ' Reaction with isocyanates
Figure imgf000086_0003
wherein Rl and R2 are as defined in claim 1 and n = 0- 4.
A method for making a compound library according to claim 4, which method comprises the step of synthesising compounds of formula I according to the following reaction scheme :
Figure imgf000087_0001
(II) (HI) i) H+ ii) NaOH(aq)
Figure imgf000087_0002
Acylation Alkylation t* Reaction with isocyanates
Figure imgf000087_0003
wherein Rl and R2 are as defined in claim 1 and n = 0-
A method for making a compound library according to claim 4 or 5 , which method comprises the step of synthesising compounds of formula I by further derivatising Rl by reaction with boronic acids from List 6.
Intermediate compounds of formula (IV) , as defined in claims 4-5, for use in a method according to any of claims 4 to 5 for making a compound library according to any of claims 1 to 3.
A compound capable of binding to a G-protein coupled receptor, which compound is selected from compounds represented within a library according to claim 1 but not including compounds having any of the following eight structures :
Figure imgf000088_0001
Figure imgf000088_0002
Figure imgf000089_0001
9. A method of making a compound according to claim 8 , which method is according to the methods of claims 4 to 6.
10. A compound library comprising or consisting of a set of structurally related compounds, substantially as herein described.
11. A compound capable of binding to a G-protein coupled receptor, substantially as herein described.
PCT/GB2004/002533 2003-06-13 2004-06-11 Compound libraries of spiro-benzodioxine-piperidines WO2004111056A2 (en)

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Publication number Priority date Publication date Assignee Title
US7772271B2 (en) 2004-07-14 2010-08-10 Ptc Therapeutics, Inc. Methods for treating hepatitis C
US7781478B2 (en) 2004-07-14 2010-08-24 Ptc Therapeutics, Inc. Methods for treating hepatitis C
US7868037B2 (en) 2004-07-14 2011-01-11 Ptc Therapeutics, Inc. Methods for treating hepatitis C
US7973069B2 (en) 2004-07-14 2011-07-05 Ptc Therapeutics, Inc. Methods for treating hepatitis C
US8013006B2 (en) 2004-07-14 2011-09-06 Ptc Therapeutics, Inc. Methods for treating hepatitis C
US7645881B2 (en) 2004-07-22 2010-01-12 Ptc Therapeutics, Inc. Methods for treating hepatitis C
WO2006067224A2 (en) * 2004-12-23 2006-06-29 Biovitrum Ab (Publ) Spiro-benzodioxole and spiro-benzodioxane compounds as orexin receptor antagonists
WO2006067224A3 (en) * 2004-12-23 2006-11-02 Biovitrum Ab Spiro-benzodioxole and spiro-benzodioxane compounds as orexin receptor antagonists
US20100298332A1 (en) * 2008-01-30 2010-11-25 Cephalon, Inc. Substituted Spirocyclic Piperidine Derivatives as Histamine-3 (H3) Receptor Ligands
US8524713B2 (en) * 2008-01-30 2013-09-03 Cephalon, Inc Substituted spirocyclic piperidine derivatives as histamine-3 (H3) receptor ligands
WO2010103381A1 (en) * 2009-03-13 2010-09-16 Glenmark Pharmaceuticals S.A. Spirocyclic piperidine derivatives as trpm 8 modulators

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