CZ20022110A3 - Peptide beta-rotating mimetic compounds and process of their preparation - Google Patents

Abstract

The subject invention involves compounds having structure (I), wherein: (a) R1 is hydrogen or alkyl; and R2 is selected from hydrogen, alkyl, aryl, heterocyclyl, carboxy and its esters and amides; or R1 and R2 are attached and are together alkylene or heteroalkylene; (b) R4 is selected from aryl, heteroaryl, and alpha , beta -unsaturated conjugated aryl or heteroaryl; and (c) R5 is selected from hydrogen, alkyl, aryl, and heterocyclyl; and an optical isomer, diesteriomer, or enantiomer thereof; a salt, hydrate, ester, amide or imide thereof. The subject invention also includes libraries of such compounds, and processes for making the subject compounds and libraries.

Description

The present invention relates to novel peptide β-rotating mimetic compounds and methods for their preparation.

Link

This application claims priority under Title 35, United States Code 119 (e) of Provisional Application Serial No. 60/172 823, filed December 21, 1999.

BACKGROUND OF THE INVENTION

In peptides, β-rotation belongs to the set of reverse rotation, which is a common characteristic of biologically active peptides and proteins. It is generally believed to act as a molecularly identifiable site for many biological processes. Specific types of β-rotations are classified according to their geometry.

β-rotation is defined as any tetrapeptide sequence with an 10-cell intramolecular H-bonded ring in which the distance between C _and 'and C _and ^{1 + 3} ranges from 0.4 to 0.7 nm (4 to 7 Å) .

i + 1

Figure 1

Depending on φ ₂ , ψ ₂ , <j> 3 and ψ3, there are many types of β-rotating structures described in the literature (see Gillespie et al., Conformational Analysis of Dipeptide Mimetics, Biopoly, Vol. 43, (1997), pp. 191-217, Venkatachalam, Biopolymers, Vol. 6, (1968), pp. 1425-36.

The compounds of the invention are mimetics for β-rotating peptides. Such compounds are useful as probes for studying molecular identification events, including enzyme inhibition, cell-cell interactions, and cell-matrix interactions. One of the physical consequences of such structural constraints on the compounds of the invention is to reduce the number of available structural states of the molecules, resulting in a better definition of the bioactive structure of the corresponding active peptides.

Information regarding β-rotating peptides and mimetic compounds can also be found, for example, in the following references: Balí et al., Β-Turn Topography, Tetrahedron, Vol. 49, no. 17 (1993), s. 3467-3478; Kahn, Peptide Secondary Structure Mimetics: Recent Advances and Future Challenges, SYNLETT, (Nov. 1993), pp. 821-826; Hanessian et al., Design and Synthesis of Conformationally Constrained Amino Acids as Versatile Scaffolds and Peptide Mimetics. Tetrahedron, Vol. 53, No. 38 (1997), s. 12789-12854.

SUMMARY OF THE INVENTION

The invention relates to compounds of formula I,

o • · · · · ·

9 9 where

a) R 1 is hydrogen or alkyl and R 2 is selected from the group consisting of hydrogen, alkyl, aryl, heterocyclyl, carboxy and esters and amides thereof, or R 1 and R 2 are taken together to form alkylene or heteroalkylene;

b) R 4 is selected from the group consisting of aryl, heteroaryl and α, β-unsaturated conjugated aryl or heteroaryl,

c) R 5 is selected from the group consisting of hydrogen, alkyl, aryl and heterocyclyl, and an optical isomer, diasteromer or enantiomer thereof, and a salt, hydrate, ester, amide, or imide thereof.

The invention also relates to a series (libraries) of such compounds and methods for preparing the compounds of the invention and methods for preparing the series.

Detailed description of the invention

Unless otherwise specified, the term "alkyl" means a hydrocarbon chain that is branched, linear or cyclic, saturated or unsaturated (but not aromatic), substituted or unsubstituted. The term "alkyl" may be used alone or as part of another word where it is optionally abbreviated to "alk" (eg, alkoxy, alkylacyl). Preferred linear alkyl contains one to twenty carbon atoms, more preferably one to ten carbon atoms, even more preferably one to six carbon atoms, even more preferably one to four carbon atoms, most preferably methyl or ethyl. Preferred cyclic and branched alkyls contain three to twenty carbon atoms, more preferably three to ten carbon atoms, even more preferably three to seven carbon atoms, even more preferably three to five carbon atoms. A preferred cyclic alkyl comprises one hydrocarbon ring, optionally two, three or more fused or spirocyclic hydrocarbon rings. Preferred unsaturated alkyls contain one to three double or triple bonds, preferably double bonds, more preferred are mono-unsaturated alkyls with one double bond. Even more preferred are saturated alkyls. Saturated alkyl is referred to herein as "alkanyl". Unsaturated alkyl with one or more double bonds (not triple bonds) is referred to herein as "alkenyl". Preferred substituents include halogen, alkyl, aryl, heterocycle, hydroxy, alkoxy, aryloxy, thio, alkylthio, arylthio, amino, alkylamino and arylamino, furthermore amide, alkylamide, arylamide, formyl, alkylacyl, arylacyl, carboxy and its alkyl and aryl esters and amides, and nitro and cyano. Also preferred are unsubstituted alkyls.

The term heteroatom means a nitrogen, oxygen or sulfur atom.

The term alkylene means alkyl that connects two other groups, the term "heteroalkylene" means alkylene with one or more heteroatoms in the linker chain.

Unless otherwise specified, the term "aryl" means an aromatic hydrocarbon ring (or fused rings) which is substituted or unsubstituted. The term "aryl" may be used alone or as part of another word (eg, aryloxy, arylacyl). A preferred aryl comprises six to fourteen, preferably up to ten, carbon atoms in the aromatic ring (s) and a total of six to twenty, preferably up to twelve, carbon atoms. Preferred aryl is phenyl or naphthyl, most preferably phenyl. Preferred substituents include halogen, alkyl, aryl, heterocycle, hydroxy, alkoxy, aryloxy, thio, alkylthio, arylthio, amino, alkylamino and arylamino, amide, alkylamide, arylamide, formyl, alkylacyl, arylacyl, carboxy and its alkyl groups and aryl esters and amides, and a nitro and cyano group. Also preferred are unsubstituted aryl.

Unless otherwise specified, the term "heterocycle" or "heterocyclyl" means a saturated, unsaturated or aromatic cyclic hydrocarbon ring (or fused rings) with one or more heteroatoms in the hydrocarbon ring (s). Preferred heterocycles contain one to six heteroatoms in the ring (s), more preferably one or two or three heteroatoms in the ring (s). Preferred heterocycles contain three to fourteen, preferably up to ten carbon atoms plus ring heteroatoms, more preferably three to seven, even more preferably five or six carbon atoms plus ring heteroatoms, and further comprising a total of three to twenty carbon atoms plus heteroatoms, more preferably three to ten, even more preferably five or six carbon atoms plus heteroatoms. Preferred heterocycles contain one ring, optionally two, three or more fused or spirocyclic rings. More preferred heterocycles contain one ring of 5 or 6 carbon atoms plus heteroatoms, of which a maximum of three are. Heteroatoms, of which no more than two are O and S. Even more preferred are those heterocycles which are formed by a ring containing 5 or 6 atoms, one or two of which are O or S and the other are C, or of which one, two or three atoms are N and the rest are C. Such preferred heterocycles, which are formed by a ring containing 5 or 6 atoms, are preferably saturated, unsaturated with one or two double bonds or aromatic. Such heterocycles which are formed by a ring containing 5 or 6 atoms are preferably single or further condensed with hydrocarbon rings of 3 to 6 atoms and which are saturated, unsaturated with one double bond or aromatic (phenyl), or further condensed with other heterocycles which are formed by a ring containing 5 or 6 atoms. Heterocycles are unsubstituted or substituted. Preferred substituents include those set forth in the definition of alkyl.

Unless otherwise specified, the term "heteroaryl" means an aromatic heterocycle.

Compounds of the invention

The present invention provides compounds of formula I

In formula I, R 1 is hydrogen or alkyl. R 1 is preferably alkyl having 1 to 12 carbon atoms, more preferably 1 to 6 carbon atoms, even more preferably 1 to 2 carbon atoms. R 1 is preferably unsubstituted or substituted alkyl, preferred substituents include aryl, heterocyclyl, amino, alkylamino, arylamino, hydroxy, alkoxy, aryloxy, thio, alkylthio and arylthio, carboxy and esters and amides thereof; further preferred substituents include phenyl, naphthyl and heterocyclyl having one ring of 5 or 6 atoms, of which 1 to 3 are heteroatoms, or further heterocyclyl having two fused ring forms 8 to 10 atoms, of which 1 to 4 are heteroatoms. More preferably, R 1 is hydrogen.

In Formula I, R 2 is selected from the group consisting of hydrogen, alkyl, aryl, heterocyclyl, carboxy, and esters and amides thereof. R 2 is preferably alkyl of 1 to 8 carbon atoms, more preferably 1 to 4 carbon atoms, even more preferably 1 to 2 carbon atoms. R 2 is preferably unsubstituted or substituted alkyl, preferred substituents include aryl, heterocyclyl, amino, alkylamino, arylamino, hydroxy, alkoxy, aryloxy, thio, alkylthio and arylthio, halogen, nitro and cyano, carboxy and its groups esters and amides; further preferred substituents include phenyl, naphthyl and heterocyclyl having one ring of 5 or 6 atoms of which 1 to 3 are heteroatoms, or further heterocyclyl having two fused rings of 8 to 10 atoms of which 1 to 4 are heteroatoms, furthermore hydroxy , -C ₆ alkoxy, phenoxy, thio, _Ci-C6 alkylthio, phenylthio, carboxy and its C1-C6 esters and amides. Aryl R2 is preferably phenyl or naphthyl, more preferably phenyl. R2 is preferably unsubstituted or substituted aryl, with preferred substituents including alkyl, aryl, heterocyclyl, amino, alkylamino, arylamino, hydroxy, alkoxy, aryloxy, thio, alkylthio and arylthio, halogen, nitro and cyano, carboxy and esters and amides thereof; more preferred substituents include C 1 -C 6 alkyl. R 2 is preferably a heterocycle with one ring of 5 or 6 atoms, of which 1 to 3 are heteroatoms, or further a heterocyclyl with two fused rings of 8 to 10 atoms, of which 1 to 4 are heteroatoms. More preferably, the heterocycle R2 is heteroaryl. R 2 is preferably an unsubstituted or substituted heterocycle, preferred substituents include alkyl, aryl, heterocyclyl, amino, alkylamino, arylamino, hydroxy, alkoxy, aryloxy, thio, alkylthio and arylthio, halogen, nitro and cyano, carboxy and esters and amides thereof; more preferred substituents include C _1-6 alkyl, phenyl, naphthyl and heterocyclyl having one ring of 5 or 6 atoms of which 1 to 3 are heteroatoms, or further heterocyclyl with two fused rings of 8 to 10 atoms of which 1 to 4 are heteroatoms .

• ·. ··· ··· · · · ··· ·· ··

Preferably, R 2 is selected from the group consisting of known α-amino acid side chains, especially those α-amino acids usually found in nature.

In formula I, R 1 and R 2 are optionally joined, and such R 1 / R 2 bonds form an alkylene or heteroalkylene. R 1 / R 2 is preferably alkylene having 1 to 6 carbon atoms, more preferably 2 to 4 carbon atoms, even more preferably 3 or 4 carbon atoms. R 1 / R 2 is preferably heteroalkylene containing 1 to 5 carbon atoms, more preferably 1 to 4 carbon atoms, even more preferably 2 or 3 carbon atoms, and further preferably 1 to 3 heteroatoms, more preferably 1 or 2 heteroatoms, even more preferably 1 heteroatom. R 1 / R 2 is preferably unsubstituted or substituted alkylene and heteroalkylene, preferred substituents of the carbon atom include alkyl, aryl, heterocyclyl, amino, alkylamino, arylamino, hydroxy, alkoxy, aryloxy, thio, alkylthio and arylthio, carboxy and esters thereof and amides; further preferred carbon atom substituents include hydroxy, Ο-β-β alkoxy, phenoxy, thio, C 1 -C ₆ alkylthio and phenylthio, carboxy and its _{C 1-6} esters and amides, preferred nitrogen substituents further include C 1 -C ₆ alkyl (unsubstituted or substituted).

In Formula I, R 4 is selected from the group consisting of aryl, heteroaryl and α, β-unsaturated conjugated aryl or heteroaryl. Aryl R4 is preferably phenyl or naphthyl, more preferably phenyl. R 4 is preferably unsubstituted or substituted aryl, with preferred substituents including alkyl, aryl, alkoxy, aryloxy, alkylthio and arylthio, halogen, nitro and cyano, carboxy and esters and amides thereof; more preferred substituents further include a C 1 -C ₆ alkyl, phenyl, C 1 -C 6 alkoxy group, and halogen. R 4 is preferably heteroaryl having one ring of 5 or 6 atoms, of which 1 to 3 are heteroatoms, or further two fused rings of 8 to 10 atoms, of which 1 to 4 are heteroatoms. R 4 is preferably unsubstituted or substituted heteroaryl, preferred substituents include alkyl, aryl, alkoxy, aryloxy, alkylthio and arylthio, halogen, nitro and cyano, carboxy and esters and amides thereof; More preferred substituents also include C1-C6 alkyl, phenyl, _Ci-C6 alkoxy and halogen. The conjugated aryl and heteroaryl R 4 is preferably styryl. R 4 is preferably unsubstituted or substituted conjugated aryl and hetroaryl, preferred substituents include alkyl, alkoxy, aryloxy, alkylthio and arylthio, halogen, nitro and cyano, carboxy and esters and amides thereof; more preferred substituents further include C1-C6 alkyl, phenyl, C1- _C6 alkoxy and halogen.

In Formula I, R 5 is selected from the group consisting of hydrogen, alkyl, aryl and heterocyclyl. R 5 is preferably alkyl having 1 to 10 carbon atoms, more preferably 1 to 4 carbon atoms, even more preferably 1 or 2 carbon atoms. R 5 is preferably unsubstituted or substituted alkyl, preferred substituents include alkyl, aryl, heterocyclyl, amino, alkylamino, arylamino, hydroxy, alkoxy, aryloxy, thio, alkylthio and arylthio, furthermore formyl, alkylacyl, arylacyl, halogen, and the like; nitro and cyano, carboxy and its esters and amides; more preferred substituents further include phenyl, naphthyl and heterocyclyl having one ring of 5 or 6 atoms, of which 1 to 3 are heteroatoms, or two fused rings of 8 to 10 atoms, of which 1 to 4 are heteroatoms. Aryl R 5 is preferably phenyl or naphthyl, more preferably phenyl. R 5 is preferably unsubstituted or substituted aryl, with preferred substituents including alkyl, aryl, heterocyclyl, amino, alkylamino, arylamino, hydroxy, alkoxy, aryloxy, thio, alkylthio and arylthio, furthermore formyl, alkylacyl, arylacyl, halogen, nitro and cyano, carboxy, and esters and amides thereof; more preferred substituents further include C 1 -C 6 alkyl, phenyl and heterocyclyl having one ring of 5 or 6 atoms of which 1 to 3 are heteroatoms, or two fused rings of 8 to 10 atoms of which 1 to 4 are heteroatoms. R 5 is preferably a heterocycle having one ring of 5 or 6 atoms, of which 1 to 3 are heteroatoms, or further two fused rings of 8 to 10 atoms, of which 1 to 4 are heteroatoms. More preferably, the heterocycle R 5 is heteroaryl. R 5 is preferably an unsubstituted or substituted heterocycle, preferred substituents include alkyl, aryl, heterocyclyl, further amino, alkylamino, arylamino, hydroxy, alkoxy, aryloxy, thio, alkylthio and arylthio, furthermore formyl, alkylacyl, arylacyl, halogen; nitro and cyano, carboxyamide, esters and amides thereof; more preferred substituents further include C 1 -C 6 alkyl, phenyl and heterocyclyl having one ring of 5 or 6 atoms of which 1 to 3 are heteroatoms, or two fused rings of 8 to 10 atoms of which 1 to 4 are heteroatoms.

The present invention relates to the optical isomers, diasteromers and enantiomers of the compounds of formula I. The invention relates to the salts, hydrates, esters, amides and imides of such compounds.

The term "salt" means a cationic salt formed on any acidic group (eg a carboxy group) or an anionic salt formed on any basic group (eg an amino group) in a compound of formula I. Many salts are known. Preferred cationic salts include alkali metal salts such as sodium or potassium salts, alkaline earth metal salts such as magnesium and calcium, and organic salts such as ammonium. Preferred anionic salts include halides, sulfonates, carboxylates, phosphates, and the like. The addition salts optionally provide an optical center that has not been there before.

The compounds of the invention and their salts optionally contain one or more chiral centers. The present invention relates to all optical isomers of a compound of formula I and salts thereof including diastereomers and enatiomers.

The invention also relates to a series of compounds of formula I. Such series are optionally mixtures of compounds of formula I or a collection of individual compounds of formula I.

4 4 · · · · · · · · · · · · · · · · · · · · · ·

5 · · »

Methods for preparing compounds

Further, the present invention relates to a process for the preparation of compounds of formula I (product 2) as generally depicted in Figure 2.

Figure 2

Orthogonally protected, resin-bound piperazine acid is used. The deprotection reaction is followed by functionalization of the β-nitrogen atom via the Petasis reaction. The subsequently formed amide bond results in the desired inclusion of R4 and R5 substituents. Unblocking α-nitrogen, followed by coupling of the Boc-N-protected (Boc-butoxycarbonyl) α-amino acid, deprotection and cyclizative cleavage, incorporates the substituents R 2 and R 1, resulting in the bicyclic product 2.

In step a, the hydroxymethylpolystyrene resin is reacted with the N-protected piperazineic acid (3), resulting in its binding to the resin. This reaction is preferably carried out in tetrahydrofuran solvent (THF) after the resin swells

· <

• ». • '*' • * * • · ♦ •. In dichloromethane (DCM), in the presence of triphenylphosphine (Ph 3 P) and diethyl azodicarboxylate (DEAD), which act as reagents for the Mitsunobu reaction. The resin product is preferably filtered and washed several times using one or more THF, DCM and methanol (MeOH) to purify the product.

In step b, the protecting group is removed from the resin-bonded β-nitrogen of piperazineic acid to give product 4. This is preferably achieved using a solution of trifluoroacetic acid (TFA) and DCM, preferably 40% TFA, preferably after swelling of the reacting resin component in DCM. The resin product is preferably filtered and washed several times using DCM and / or MeOH, neutralized and again filtered and washed for purification.

In step c, the piperazine ester of resin 4 is reacted with R4-boronic acid and glyoxylic acid. This reaction is preferably carried out in a DCM solution, preferably after swelling of the reacting resin component in DCM. The resin product is preferably filtered and washed several times using DCM and MeOH, resulting in purification. This reaction step and purification are preferably repeated to increase the yield of the desired product.

In step d, the resin product of step c is reacted with an amine R 5 -NH ₂ . Prior to the addition of the amine in this step, the reacting resin component is preferably swelled in dimethylformamide (DMF), mixed with hydroxybenzotriazole (HOBt), which acts as an activating agent, then mixed with diisopropylcarbodiimide (DIC), which acts as coupling agent, and further filtered and washed with DMF. After reaction with the amine, the resin product is preferably filtered and washed with DMF.

In step e, the resin product of step d is mixed with piperidine, which acts as a deprotecting agent. This step is preferably carried out in a solution of piperidine in DMF containing 15 to 30% piperidine. The resulting resin product 5 is preferably filtered and washed several times with DMF. This step is preferably repeated to increase the yield of the desired product.

• 0

In step f, the resin product 5 is reacted with an N-protected α-amino acid. This reaction is preferably carried out in the presence of benzotriazol-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate (PyBOP), which acts as a coupling agent and then in the presence of a base of diisopropylethylamine (DiPEA). Before the amino acid is added, the reacting resin component is preferably swelled in DMF. The resin product is preferably filtered and washed several times with DMF. This step is preferably repeated.

In step g, the α-amino acid protecting group is removed from step f. This is preferably done using a solution of TFA in DCM, preferably 25% TFA. The resin product is preferably filtered and washed several times with DMF and MeOH.

In step h, the resin product of step g is cleaved from the resin and cyclized to give product 2. This step is preferably carried out in a solution of 5 to 20% acetic acid (AcOH) in isopropanol (iPrOH) at an elevated temperature of 30 to 80 ° C for The resolved resin is filtered and washed several times, preferably using MeOH. The filtrate and washings are preferably mixed, concentrated and dried to give solid products 2. The solid product is preferably purified by repeated evaporation with chloroform and then drying under vacuum.

♦

DETAILED DESCRIPTION OF THE INVENTION

The following non-limiting examples illustrate the preparation of the compounds of this invention in more detail.

Example 1

a) Hydroxymethylpolystyrene resin (1.0 g, 1.44 mmol / g, Advanced Chemtech) was swelled in anhydrous DCM (6 mL). Ph3P (1.13 g, 4.32 mmol) was dissolved in this suspension and the heterogeneous reaction was cooled to 0 ° C with nitrogen. To this finely stirred suspension was added THF (30 mL) solution of N-Fmoc-Np-Boc-2-carboxy-piperazine 3 (Fmoc-fluorenylmethyloxycarbonyl, Np-nitropiperonyl) (1.95 g, 4.32 mmol) over 30 minutes. ) and DEAD (751 mL, 4.32 mmol). The mixture was stirred for 72 h after which time the resin was filtered and washed with THF (3x), DCM (3x), MeOH (3x) and then washed several times, successively and alternately with DCM and MeOH (standard method).

b) The piperazine ester of the resin of step a (1.4 g) was swelled in DCM, filtered and purified for 1 h with a 40% solution of TFA in DCM. The resin was filtered and washed with DCM (3x), MeOH (3x) and then alternately with DCM / MEOH as described in step (a). The resin-bound amine TFA is then neutralized with a 10% solution of DiPEA in DCM and washed again in a standard manner.

c) The resin ester 4 from step b was swelled in DCM (10 mL) and glyoxylic acid (265 g, 2.88 mmol) and boronic acid (2.88 mmol) were added as a solution in MeOH (15 mL). The resulting suspension was stirred for 5 h before filtering the resin and washing the MeOH (3X). The procedure described above was repeated for 16 h, after which time the resin was filtered again and washed in a standard manner.

d) The resin ester from step c was swelled in DMF (5 mL) and HOBt (1.10 g, 7.2 mmol) was added followed by DIC (907 mg, 7.2 mmol). The mixture was stirred for 3 h, after which time the resin was filtered and washed with DMF (4x). Resin

•

was re-swelled in DMF (10 mL) and amine (7.2 mmol) was added to this suspension. The mixture was stirred for 15 h. The resin was filtered and washed with DMF (3x) and further washed in a standard manner.

e) The resin ester of step d was purified with 25% piperidine in DMF for 20 min. The resin was filtered and washed with DMF (2x) before repeating. The resin product is filtered and washed in a standard manner.

f) The resin product 5 of step e is swollen in DMF. Add Boc-α-amino acid (Boc-AA) (7.2 mmol), PyBOP (3.74 g, 7.2 mmol) followed by DiPEA (1.1 g, 8.64 mmol). The mixture was stirred for 5 h, filtered and washed with DMF (3x) in a standard manner.

g) The resin product of step f was purified with 25% TFA / DCM for 1 h. The resin was filtered and washed.

h) The resin product of step g was mixed with 10% AcOH in iPrOH and kept at 50 ° C for 16 h. The resin was filtered and washed several times with MeOH. The filtrate and washings were combined and concentrated to give an off-white solid. The solid product was evaporated several times with chloroform and then dried under vacuum for 15 h.

with

Examples 2 to 7

In the following non-limiting examples, compounds are prepared as described in Example 1 using the boronic acid, amine and Boc-α-amino acid derivatives below instead of the original ones.

• · * «fe» ·· »♦ · * · _Λ • fe" · · · ·· • · · · · · fe • Fefe · · ·

Bfe «fe · fe fe · fe fe fe

Example Boronic acid Amin Aminoacid Product 2 acid phenylboronic acid isoamylamine Boc-Asp (O-tBu) -OH 2a 3 acid 2-thiophenoborone mono-Boc-butyl- -1,4-diamine Boc-Lys (Boc) -OH 2b 4 acid benzothiofen- -2-boron 4-aminomethyl- -pyridine Boc-Tic-OH 2c 5 acid 4-Fluoroborone phenethylamine Boc-Phe-OH 2d 6 acid 2-furylboron benzylamine Boc-Phe-OH 2e 7 acid 2-naphthylboronic acid aniline Boc-Hyp (OBn) -OH 2f

(Asp-aspartic acid) (tBu-tert-butyl) (Lys-lysine) (Tic-tetrahydroisoquinoline-3-carboxylic acid) (Phe-phenylalanine) (Hyp-4-hydroxyproline) (OBn-alkaloid ouabain)

The above-described processes for preparing the compounds of this invention are carried out using a solid-forming resin. This allows the reaction, separation and purification of the intermediate and end product to be automated. A series of compounds of formula I are readily prepared using the preparation methods of this invention. The automation of the preparation of such series is achieved by the use of equipment known to those skilled in the art. Such a device can be used to prepare series, which are mixtures of compounds of formula I or a collection of individual compounds of formula I, each in an isolated chamber.

»444 ··

4 »··

4 · ♦

4 4

4 4 4

4 * 4

4444

Although particular embodiments of the present invention have been described, it will be apparent to those skilled in the art that they can be made with various changes and modifications without departing from the spirit and scope of the present invention. All such modifications that do not depart from the scope of the invention are included in the following claims.

Industrial applicability

The compounds of this invention are useful as probes for studying molecular identification events.

Claims (10)

PATENT CLAIMS A compound of formula I wherein (a) R 1 is hydrogen or alkyl and R 2 is selected from the group consisting of hydrogen, alkyl, aryl, heterocyclyl, carboxy and esters and amides thereof, or R 1 and R 2 are optionally joined to form alkylene together or heteroalkylene, (b) R 4 is selected from the group consisting of aryl, heteroaryl and α, β-unsaturated conjugated aryl or heteroaryl, (c) R 5 is selected from the group consisting of hydrogen, alkyl, aryl and heterocyclyl, and an optical isomer, diasteromer thereof or an enantiomer, a salt, a hydrate, an ester, an amide or an imide thereof. A compound according to claim 1, wherein (a) R 1 is hydrogen or C 1 -C 12 alkyl and R 2 is selected from the group consisting of hydrogen, C 1 -C 8 alkyl, phenyl, naphthyl and heterocyclyl having one ring of 5 or 6 atoms of which 1 to 3 are heteroatoms, or further two fused rings of 8 to 10 atoms, of which 1 to 4 are heteroatoms, or R1 and R2 are joined together to form C1-C6 alkylene or heteroalkylene of 1 to 5 carbon atoms and 1 to 3 heteroatoms, ( b) R 4 is selected from the group consisting of phenyl, naphthyl, styryl and hetroaryl having one ring of 5 or 6 atoms of which 1 to 3 are heteroatoms or two fused rings of 8 to 10 atoms of which 1 to 4 are heteroatoms , • · (c) R5 is selected from the group consisting of hydrogen, _Ci-Cio alkyl, phenyl, naphthyl, and heterocyclyl having one ring formed of 5 or 6 atoms of which 1-3 are heteroatoms, or two condensed rings containing 8 to 10 carbon of which 1 to 4 are heter oatomy. The compound of claim 2, wherein (a) R 1 is hydrogen or unsubstituted or substituted C 1-6 alkyl and R 2 is selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 4 alkyl, unsubstituted or substituted phenyl, unsubstituted or substituted heteroaryl with one ring of 5 or 6 atoms, of which 1 to 3 are heteroatoms, or R 1 and R 2 are joined together to form unsubstituted or substituted C 1 -C 4 alkylene or further unsubstituted or substituted C 1 -C 4 heteroalkylene also containing one or two heteroatoms, ( b) R 4 is selected from the group consisting of unsubstituted or substituted phenyl, unsubstituted or substituted styryl and unsubstituted or substituted heteroaryl having one ring of 5 or 6 atoms of which 1 to 3 are heteroatoms, (c) R 5 is selected from hydrogen, unsubstituted or substituted C 1 -C 4 alkyl, unsubstituted or substituted phenyl; and unsubstituted or substituted heterocyclyl having one ring of 5 or 6 atoms, of which 1 to 3 are heteroatoms. A compound according to claim 3, wherein (a) the R 1 alkyl substituents are phenyl or a single ring of 5 or 6 atoms of which 1 to 3 are heteroatoms, (b) the R 2 alkyl substituents are selected from phenyl, heterocyclyl with one ring of 5 or 6 atoms of which 1 to 3 are heteroatoms, hydroxy, C 1 -C ₆ alkoxy, phenoxy, thio, C 1 -C 6 alkylthio and phenylthio, carboxy and its C 1 -C ₆ esters and amides, (c) aryl substituents R2 is C1- _C6 alkyl, (d) the heterocyclyl substituents R2 are selected from the group consisting of C1-C6 alkyl, phenyl and heterocyclyl having one ring of 5 or 6 atoms, of which 1 to 3 are heteroatoms, (e) ) the R 1 / R 2 alkylene and heteroalkylene substituents are selected from the group consisting of C _1-6 alkyl, hydroxy, C 1 -C ₆ alkoxy, phenoxy, thio, C 1 -C 6 alkylthio and phenylthio, carboxy and its _{C 1-6} esters and amides; the phenyl, heteroaryl and styryl substituents R 4 are selected from the group consisting of obs exceeding -C ₆ alkyl, phenyl, _Ci-C6 alkoxy and halogen, (g) alkyl substitutents R 5 are phenyl or heterocyclyl having one ring formed of 5 or 6 atoms of which 1-3 are heteroatoms, (h) substituents of aryl and heterocyclyl R 5 is selected from the group consisting of C _1-6 alkyl, phenyl and heterocyclyl having one ring of 5 or 6 atoms, of which 1 to 3 are heteroatoms. The compound of claim 3, wherein R 1 is hydrogen or alkyl, R 1 and R 2 are linked and R 4 is aryl, heteroaryl or α, β-unsaturated conjugated aryl or heteroaryl. 6. A process for the preparation of a compound of formula I, wherein the piperazine ester or resin ester is used as reactants. ··· · ·· · ·· · The method of claim 8, wherein the β-nitrogen atom of the piperazine ester is functionalized by a Petasis reaction of boronic acid, wherein (a) the hydroxymethylpolystyrene resin reacts with the N-protected piperazine acid resulting in its binding to the resin to form the product in step (a), (b) the protecting group is removed from the β-nitrogen of the product of step (a), resulting in the product of step (b), (c) the product of step (b) reacts with R4-boronic acid and acid glyoxyl, resulting in the product of step (c), ^with (d) the product of step (c) reacting with the amine R 5 -NH ₂ , resulting in the product of step (d), (e) removing from the product of step (d) protecting group resulting in the product of step (e), (f) the product of step (e) reacts with the N-protected α-amino acid, resulting in the product of step (f), (g) removing the α-amino acid protecting group the product of step (f), resulting in the product of step (g), (h) the product of kr Eye (g) is cleaved from the resin and cyclized to give a compound. A process according to claim 9, characterized in that (a) in step (c) the product of step (b) is swelled in DCM and the reaction is carried out in a solvent of MeOH, * (b) in step (b). d) the product of step (c) is swollen in DMF, contacted with HOBt, which acts as an activating agent, and contacted with DIC, which acts as a coupling agent, (c) in step (f), the product of step (e) swell in DMF, contact with PyBOP, which acts as coupling agent, and DiPEA, and then contact with amino acid, (d) in step (h) use a 5-20% solution of AcOH in iPrOH. tt ·· A series of compounds wherein any one of claims 1, 3, 4, 8 or 9. A series of compounds characterized by the method of claim 9. contain the compounds according to