WO1997033174A1 - Selectively n-alkylated peptidomimetic combinatorial libraries and compounds therein - Google Patents
Selectively n-alkylated peptidomimetic combinatorial libraries and compounds therein Download PDFInfo
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- WO1997033174A1 WO1997033174A1 PCT/IB1997/000349 IB9700349W WO9733174A1 WO 1997033174 A1 WO1997033174 A1 WO 1997033174A1 IB 9700349 W IB9700349 W IB 9700349W WO 9733174 A1 WO9733174 A1 WO 9733174A1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
- C07K1/047—Simultaneous synthesis of different peptide species; Peptide libraries
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
Definitions
- the present invention relates generally to novel, selectively N-alkylated compounds of Formula I below, as well as novel libraries composed of many such compounds, methods of synthesizing and screening the libraries, and methods of using the compounds.
- the process of discovering new therapeutically active compounds for a given indication involves the screening of all compounds from available compound collections. From the compounds tested, one or more structure (s) is selected as a promising lead. A large number of related analogs are then synthesized in order to develop a structure-activity relationship and select one or more optimal compounds. With traditional one-at- a-time synthesis and biological testing of analogs, this optimization process is long and labor intensive. Adding significant numbers of new structures to the compound collections used in the initial screening step of the discovery and optimization process cannot be accomplished with traditional one-at-a-time synthesis methods, except over a time frame of months or even years. Faster methods are needed that allow for the preparation of up to thousands of related compounds in a matter of days or a few weeks. This need is particularly evident when it comes to synthesizing more complex compounds, such as the instant compounds composed of two or more monomers, each monomer possessing more than one variable substituent.
- SCLs chemically synthesized combinatorial libraries
- the preparation and use of synthetic peptide combinatorial libraries has been described, for example, by Dooley in U.S. Patent 5,367,053, Huebner in U.S. Patent 5,182,366, Appel et al. in WO PCT 92/09300, Geysen in published European Patent Application 0 138 855 and Pirrung in U.S. Patent 5,143,853.
- SCLs provide the efficient synthesis of an extraordinary number of various peptides in such libraries and the rapid screening of the library which identifies lead pharmaceutical peptides.
- the present invention combines the techniques of solid-phase synthesis of peptidomimetic compounds and the general techniques of synthesis of combinatorial libraries to prepare new selective
- N-alkylated compounds N-alkylated compounds.
- This invention is directed to a single selectively N-alkylated compound or a library of an approximately equimolar mixture of two more selectively N-alkylated compounds of the Formula (I):
- R 1 and R 2 independently are a hydrogen atom, an amino protecting group, C 1 to C 12 acyl, C 3 to C 10
- R 3 , R 5 , and R 7 are independently a hydrogen atom
- R 4 , R 6 and R 8 are independently a C 1 to C 18 substituent group; with the proviso that all but one of R 4 , R 6 and R 8 can simultaneously be the same group;
- R 9 is a hydrogen atom or a solid support
- R 10 is optionally present as a C 1 to C 18 substituent group when R 1 and R 2 are other than a hydrogen atom, an amino protecting group or when both R 1 and R 2 are C 1 to C 12 acyl groups;
- AA, BB, and CC are independently 0 to 5;
- B is from 0 to 3 ; further wherein the stereochemistry at the carbons bonded to R 3 , R 5 , and R 7 are independently R or S or a mixture of the two; further wherein when B is 2 or 3; each R 4 and R 5 can be the same or different; with the proviso that either R 1 or R 2 can be taken with R 3 ; R 4 can be taken with R 5 ; and R 6 can be taken with R 7 ; respectively and independently, to form a subtituted or unsubstituted pyrrolidine ring;
- X and Y are either 1) each a hydrogen atom or 2) taken together to represent a carbonyl group; and a pharmaceutically acceptable salt, solvate or hydrate thereof.
- This invention is also directed to iterative and positional scanning methods of synthesizing the libraries of compounds described above as discussed below.
- Another aspect of the invention is a method of selective N-alkylation as set forth below.
- the invention comprises methods for affecting analgesia in a mammal, effecting a decrease in the postprandial rise in the blood glucose levels of a mammal after said mammal has ingested a carbohydrate load, and a method for treating microbial infections, all of which methods comprise administering a single compound of the above formula in conjunction with a pharmaceutically acceptable carrier, as set forth below.
- the instant invention is directed to a single compound or an approximately equimolar mixture of two or more selectively N-alkylated compounds of the Formula (I) :
- R 1 and R 2 independently are a hydrogen atom, an amino protecting group, C 1 to C 12 acyl, C 3 to C 10
- cycloalkyl C 3 to C 6 heterocycle, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle;
- R 3, R 5, and R 7 are independently a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16
- alkylaryl a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle
- R 4 , R 6 and R B are independently a C 1 to C 18 substituent group; with the proviso that all but one of R 4 , R 6 and R ⁇ can simultaneoulsy be the same group;
- R 9 is a hydrogen atom or a solid support
- R 10 is optionally present as a C 1 to C 18 substituent group when R 1 and R 2 are other than a hydrogen atom or an amino protecting group;
- AA, BB, and CC are independently 0 to 5 ;
- B is from 0 to 3 ; further wherein the stereochemistry at the carbons bonded to R 3 , R 5 , and R 7 are independently R or S or a mixture of the two; further wherein when B is 2 or 3; each R 4 and R 5 can be the same or different; with the proviso that either R 1 or R 2 can be taken with R 3 ; R 4 can be taken with R 5 ; and R 6 can be taken with R 7 ; respectively and independently, to form a subtituted or unsubstituted pyrrolidine ring; X and Y are either 1) each a hydrogen atom or 2) taken together to represent a carbonyl group; and a pharmaceutically acceptable salt, solvate or hydrate thereof.
- the terms used in the above Formula I having the following meanings when used in conjunction with Formula I and when used in described subsequent Formulas :
- cycloalkyl - unsubstituted or substituted mono- or bicyclic saturated rings such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or adamantyl or rings wherein the
- substituents are one or more hydroxy, halo, amino, protected amino, carboxy, protected carboxy, amido, nitro, trifluoromethyl, phenyl, heterocyclic rings, C 1 to C 7 acyl, C 1 to C 7 alkoxy, or C 1 to C 3 alkyl groups;
- C 1 to C 18 Substituent Group indicates a group of the formula -CH 2 -W wherein W is chosen from the group consisting of a hydrogen atom, C 1 to C 12 alkyl, C 3 to C 10 cycloalkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16 substituted alkylaryl, as those terms are defined herein;
- C 1 to C 12 substituted alkyl denotes the above C 1 to C 12 alkyl groups that are substituted by one to three halogen, hydroxy, protected hydroxy, amino, protected amino, guanidino, C 1 to C 7 acyloxy, C 1 to C 7 acyl, nitro, carboxy, protected carboxy, carboxamide, carbonyl, carboxyl, cyano, methylsulfonylamino or C 1 to C 4 alkoxy groups.
- the substituted alkyl groups may be substituted once or twice with the same or with different substituents; Examples of the above substituted alkyl groups include the cyanomethyl, nitromethyl, hydroxymethyl, trityloxymethyl, propionyloxymethyl, aminomethyl,
- C 1 to C 6 substituted alkyl includes the substituted methyl group, in other words, a methyl group substituted by the same
- substituents as the " C 1 to C 6 substituted alkyl" group include groups such as protected hydroxymethyl, (e.g.,
- substituted phenyl specifies a phenyl group substituted with one or more, and preferably one or two, moieties chosen from the groups consisting of halogen, hydroxy, protected hydroxy, cyano, nitro, trifluoromethyl, C 1 to C 6 alkyl, C 1 to C 7 alkoxy, C 1 to C 7 acyl, C 1 to C 7 acyloxy, carboxy, protected carboxy, carboxymethyl, protected carboxymethyl, hydroxymethyl, protected hydroxymethyl, amino, protected amino,
- substituted phenyl includes a mono- or di (halo) phenyl group such as 2, 3 or 4-chlorophenyl, 2, 6-dichlorophenyl, 2 , 5-dichlorophenyl, 3,4-dichlorophenyl, 2, 3 or 4-bromophenyl, 3,4- dibromophenyl, 3-chloro-4-fluorophenyl, 2, 3 or 4- fluorophenyl and the like; a mono or di (hydroxy) phenyl group such as 2, 3 or 4-hydroxyphenyl, 2,4- dihydroxyphenyl, the protected-hydroxy derivatives thereof and the like; a nitrophenyl group such as 2 , 3 or 4-nitrophenyl; a cyanophenyl group, for example, 2, 3 or 4-cyanophenyl; a mono- or di (alkyl) phenyl group such as 2, 3 or 4-methylphenyl, 2,4-dimethylphenyl, 2, 3 or 4-
- hydroxymethyl)phenyl such as 2, 3, or 4-(protected hydroxymethyl)phenyl or 3,4-di(hydroxymethyl)phenyl; a mono- or di(aminomethyl)phenyl or (protected
- aminomethyl)phenyl such as 2, 3 or 4-(aminomethyl)phenyl or 2,4-(protected aminomethyl)phenyl; or a mono- or di (N- (methylsulfonylamino))phenyl such as 2, 3 or 4-(N- (methylsulfonylamino))phenyl.
- a mono- or di (N- (methylsulfonylamino))phenyl such as 2, 3 or 4-(N- (methylsulfonylamino)phenyl.
- substituted phenyl represents disubstituted phenyl groups wherein the substituents are different, for
- halo and halogen refer to the fluoro, chloro, bromo or iodo groups. There can be one or more halogen, which are the same or different.
- C 7 to C 16 alkylaryl denotes a C 1 to C 6 alkyl group substituted at any position by a phenyl or naphthyl ring.
- Examples of such a group include benzyl, 2-phenylethyl, 3-phenyl(n-propyl), 4-phenylhexyl, 3- phenyl(n-amyl), 3-phenyl(sec-butyl) and the like.
- Preferred C 7 to C 16 phenylalkyl groups are the benzyl phenylethyl napth-1-ylmethyl and napth-2-ylmethyl groups.
- C 7 to C 16 substituted alkylaryl denotes a C 7 to C 16 alkylaryl group substituted on the C 1 to C 6 alkyl portion with one or more, and preferably one or two, groups chosen from halogen, hydroxy, protected hydroxy, oxo, protected oxo, amino, protected amino, (monosubstituted) amino, protected (monosubstituted) amino, (disubstituted) amino, guanidino, heterocyclic ring, substituted heterocyclic ring, C 1 to C 7 alkoxy, C 1 to C 7 acyl, C 1 to C 7 acyloxy, nitro, carboxy, protected carboxy, carbamoyl, carboxamide, protected carboxamide, N-(C 1 to C 6 alkyl) carboxamide, protected N-C 1 to C 6 alkyl) carboxamide, N, N-( C i to C 6 dialkyl) carboxamide, cyano, N-((C 1 to C 1 to C
- substituted alkyl or phenyl groups may be substituted with one or more, and preferably one or two, substituents which can be the same or different.
- C 7 to C 16 substituted alkylaryl include groups such as 2-phenyl-1-chloroethyl, 2-(4-methoxyphenyl)ethyl, 4-(2,6-dihydroxy phenyl)n- hexyl, 2-(5-cyano-3-methoxyphenyl)n-pentyl, 3-(2,6- dimethylphenyl) n-propyl, 4-chloro-3-aminobenzyl, 6-(4- methoxyphenyl)-3-carboxy (n-hexyl), 5-(4- aminomethylphenyl)-3-(aminomethyl) n-pentyl, 5-phenyl-3- oxo-n-pent-1-yl, (4-hydroxynapth-2-yl)methyl and the like.
- (monosubstituted) amino refers to an amino group with one substituent chosen from the group consisting of phenyl, substituted phenyl, C 1 to C 6 alkyl, C 1 to C 6 substituted alkyl, C 1 to C 7 acyl, C 2 to C 7 alkenyl, C 2 to C 7 substituted alkenyl, C 2 to C 7 alkynyl, C 7 to C 16 alkylaryl, C 7 to C 16 substituted alkylaryl and
- the (monosubstituted) amino can additionally have an amino-protecting group as
- (disubstituted) amino refers to amino groups with two substituents chosen from the group
- phenyl substituted phenyl, C 1 to C 6 alkyl, C 1 to C 6 substituted alkyl, C 1 to C 7 acyl, C 2 to C 7 alkenyl, C 2 to C 7 alkynyl, C 7 to C 16 alkylaryl, C 7 to C 16 substituted alkylaryl and heterocyclic ring.
- the two substituents can be the same or different.
- pharmaceutically-acceptable salt encompasses those salts that form with the carboxylate anions and includes salts formed with the organic and inorganic cations such as those chosen from the alkali and alkaline earth metals, (for example, lithium, sodium, potassium, magnesium, barium and calcium); ammonium; and the organic cations (for example, dibenzylammonium, benzylammonium, 2-hydroxyethylammonium,
- procaine protonated form of procaine, quinine and N-methylglucosamine
- protonated forms of basic amino acids such as glycine, ornithine, histidine, phenylglycine, lysine, and arginine
- acetic acid-like counter-ions such as acetate and trifluoroacetate .
- carboxylate anion compounds formed by a carboxylic acid and an amino group is referred to by this term.
- a preferred cation for the carboxylate anion is the sodium cation.
- the term includes salts that form by standard acid-base reactions with basic groups (such as amino groups) and organic or inorganic acids. Such acids include
- the compounds of Formula I may also exist as solvates and hydrates. Thus, these compounds may
- carboxy-protecting group refers to one of the ester derivatives of the carboxylic acid group commonly employed to block or protect the carboxylic acid group while reactions are carried out on other functional groups on the compound.
- carboxylic acid protecting groups include t-butyl, 4-nitrobenzyl, 4-methoxybenzyl, 3,4- dimethoxybenzyl, 2,4-dimethoxybenzyl, 2,4,6- trimethoxybenzyl, 2,4,6-trimethylbenzyl,
- hydroxy-protecting group refers to readily cleavable groups bonded to hydroxyl groups, such as the tetrahydropyranyl, 2-methoxyprop-2-yl,
- a preferred hydroxy-protecting group is the tert- butyl group.
- protected hydroxy denotes a hydroxy group bonded to one of the above hydroxy protecting groups.
- amino-protecting group refers to substituents of the amino group commonly employed to block or protect the amino functionality while reacting other functional groups of the molecule.
- protected (monosubstituted) amino means there is an amino-protecting group on the monosubstituted amino nitrogen atom.
- protected amino-protecting group means there is an amino-protecting group on the monosubstituted amino nitrogen atom.
- carboxamide means there is an amino-protecting group on the carboxamide nitrogen.
- amino-protecting groups include the formyl ("For") group, the trityl group, the phthalimido group, the trichloroacetyl group, the trifluoro-acetyl group, the chloroacetyl, bromoacetyl, and iodoacetyl groups, urethane-type blocking groups, such as t-butoxycarbonyl ("Boc”), 2-(4-biphenylyl)propyl- 2-oxycarbonyl ("Bpoc”), 2-phenylpropyl-2-oxycarbonyl (“Poc”), 2- (4-xenyl) isopropoxycarbonyl, 1,1- diphenylethyl-1-oxycarbonyl, 1,1-diphenylpropyl-1- oxycarbonyl, 2-(3,5-dimethoxyphenyl)propyl-2-oxycarbonyl (“Ddz”), 2-(p-1oluyl)propyl-2-oxycarbonyl,
- benzyloxycarbonyl (“Cbz”), 4-phenylbenzyloxycarbonyl, 2-methylbenzyloxy-carbonyl, ⁇ -2,4,5,- tetramethylbenzyloxycarbonyl (“Tmz”),
- PMC 2,2,5,7,8-pentamethylchroman-6-sulfonyl group
- Dts dithiasuccinoyl
- Nps 2-(nitro)phenylsulfenyl group
- diphenyl-phosphine oxide group and like amino-protecting groups.
- the species of amino- protecting group employed is not critical so long as the derivatized amino group is stable to the conditions of the subsequent reaction (s) and can be removed at the appropriate point without disrupting the remainder of the compounds.
- Preferred amino-protecting groups are Boc, Cbz and Fmoc.
- Further examples of amino-protecting groups embraced by the above term are well known in organic synthesis and the peptide art and are described by, for example, T.W. Greene and P.G.M. Wuts, "Protective Groups in Organic Synthesis," 2nd ed., John Wiley and Sons, New York, NY, 1991, Chapter 7, M. Bodanzsky,
- heterocycle denotes optionally substituted five-membered or six-membered rings that have 1 to 4 heteroatoms, such as oxygen, sulfur and/or nitrogen, in particular nitrogen, either alone or in conjunction with sulfur or oxygen ring atoms. These five-membered or six-membered rings may be saturated, fully unsaturated or partially unsaturated, with fully saturated rings being preferred.
- An "amino-substituted heterocyclic ring” means any one of the above-described heterocyclic rings is substituted with at least one amino group.
- Preferred heterocyclic rings include morpholino, piperidinyl, piperazinyl, tetrahydrofurano, pyrrolo, and tetrahydrothiophenyl.
- the above optionally substituted five-membered or six-membered rings can optionally be fused to an aromatic 5-membered or 6-membered ring system, such as a pyridine or a triazole system, and preferably to a benzene ring.
- an aromatic 5-membered or 6-membered ring system such as a pyridine or a triazole system, and preferably to a benzene ring.
- heterocyclic ring thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl,
- thiadiazolyl oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, thiazinyl, oxazinyl, triazinyl, thiadiazinyl,
- oxadiazinyl dithiazinyl, dioxazinyl, oxathiazinyl, tetrazinyl, thiatriazinyl, oxatriazinyl, dithiadiazinyl, imidazolinyl, dihydropyrimidyl, tetrahydropyrimidyl, tetrazolo[1,5-b]pyridazinyl and purinyl, as well as benzo-fused derivatives, for example benzoxazolyl, benzthiazolyl, benzimidazolyl and indolyl. Further specific examples of the above
- heterocyclic ring systems are 6-membered ring systems containing one to three nitrogen atoms.
- Such examples include pyridyl, such as pyrid-2-yl, pyrid-3-yl and pyrid-4-yl; pyrimidyl, preferably pyrimid-2-yl and pyrimid-4-yl; triazinyl, preferably 1,3,4-triazin-2-yl and 1,3,5-triazin-4-yl; pyridazinyl, in particular pyridazin-3-yl, and pyrazinyl.
- pyridine N-oxides and pyridazine N-oxides are a preferred group.
- heterocyclic ring are included in the heterocyclic thiomethyl groups listed under heading "A”.
- C 6 to C 15 alkyl heterocycle denotes a C 1 to C 6 alkyl group substituted at any position by a heterocycle ring (heterocycle) from as described above, said heterocycle containing up to 14 carbon atoms, as long as sum of the carbon atoms of the alkyl chain (up to 6) and the carbon atoms of the heterocycle do not exceed 15.
- substituted C 6 to C 1S alkyl heterocycle refer to a C 6 to C 15 alkyl heterocycle group substituted on the alkyl portion with the same substituents as listed for the C 7 to C 16 substituted alkylaryl groups and on the heterocycle as defined above for substituted heterocyle.
- a preferred group of single compounds are the interior amido compounds, that is, wherein X and Y are taken together to form a carbonyl moiety.
- interior amido single compounds are the dimers, thus, wherein B, AA, BB and CC are zero, except that AA can be zero or one when R 7 is a hydrogen atom and that CC can be zero or one when R 3 is a hydrogen atom.
- a preferred group of interior amido dimers are those that are cleaved from the solid support and are not quaternized, thus, wherein R 9 is a hydrogen atom and R 10 is absent.
- a preferred group of cleaved interior amido dimer single compounds are those wherein R 3 and R 7 are independently chosen from the group consiting of S- or R-methyl, S- or R-benzyl, a hydrogen atom, S- or R- (but-2-yl), S- or R-[ 4-(N-methylamino)-n-butyl], S- or R-[4-(N-ethylamino)-n-butyl], S- or R-[4-(N-allylamino)- n-butyl], S- or R-[4-(N-benzylamino)-n-butyl], S- or R- [4-(N-(napth-2-ylmethylamino)-n-butyl], S- or R- [4- (amino)-n-butyl], S- or R-[sec-butyl], S- or R- (methylsulfinyl) eth-1-yl, S- or R-
- Type I amido dimers compounds referred to as cleaved interior amido dimers, hereafter referred to as the "Type I" amido dimers, is wherein R 6 and R 8 are independently methyl, ethyl, allyl, benzyl, or napth-2-ylmethyl.
- a preferred group of "Type I" amido dimers are the “Type II” amido dimers, thus, wherein either R 1 or R 2 are each a hydrogen atom, or one of R 1 or R 2 is a hydrogen atom and the other is taken together with R 3 to form an S-pyrrolidine ring.
- a preferred group of the Type II interior amido dimers is the N-terminal monomer as a proline residue, thus, wherein one of R 1 or R 2 is a hydrogen atom and the other is taken together with R 3 to form an S-pyrrolidine ring.
- a preferred group of N-terminal proline Type II interior amido dimers occurs when R 6 is napth-2-ylmethyl and R 8 is benzyl, and more so when R 7 is S- or R-methyl, a hydrogen atom, S- or R-[3-(guanidino)-n-propyl], S- or R-[4-(N- benzylamino)-n-butyl], S-[iso-propyl], S-[2- (methylsulfinyl)ethyl], S- or R-(n-propyl), S- or R- (hydroxymethyl), S- or R-[n-butyl], R-[(napth-2- yl) methyl], or S-phenyl, and especially so when the C- terminal residue is S- or R-alanine, thus, R 7 is a S- or R-methyl.
- Another preferred group of Type II interior amido dimers has the N-terminal residue as a
- R 6 is ethyl and R 8 is (napth-2-yl)methyl, and especially so when R 7 is S- methyl, S-(2-(methylsulfinyl) ethyl), a hydrogen atom, S- (4-(hydroxy)benzyl) or S-[(hydroxy)methyl].
- R 7 is S-methyl.
- Type I interior amido dimers are wherein R 1 and R 2 are each a hydrogen atom and R 3 is R-[(N-(napth-2-ylmethyl)indol-3-yl)methyl].
- R 6 is napth-2-ylmethyl and R 8 is benzyl, and especially so when R 7 is S- or R-[3-(guanidino)-n-propyl] or S- or R- [4-(benzylamino)-n-butyl].
- Type I interior amido dimers occurs wherein either R 1 or R 2 is a hydrogen atom or is taken in conjunction with R 3 to form a
- pyrrolidine ring and the other is C 1 to C 12 acyl, C 3 to C 10 cycloalkyl, C 3 to C 6 heterocycle, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle.
- Another preferred group of compounds of the above Formula I wherein a single compound is indicated is the interior amine compounds, in other words, wherein X and Y are each a hydrogen atom.
- a preferred group of these interior amine compounds is dimers, wherein B, AA, BB and CC are zero, except that AA can be zero or one when R 7 is a hydrogen atom and that CC can be zero or one when R 3 is a hydrogen atom.
- a preferred group of interior amine dimers is those cleaved from the solid support, thus, wherein R 9 is a hydrogen atom and R 10 is absent.
- a preferred group of cleaved interior amine dimers is those wherein R 6 and R 8 are independently methyl, benzyl or 4-hydroxybenzyl.
- a preferred group of these preferred cleaved interior dimers are referred to
- Type I cleaved interior amine dimers that is, wherein R 3 and R 7 are independently benzyl or 4- hydroxybenzyl.
- a preferred group of the Type I cleaved interior amine dimers are those wherein R 8 is benzyl, R 7 is 4-hydroxybenzyl, R 6 is methyl, and R 3 is 4-hydroxybenzyl.
- Type I compounds are wherein a) R 1 and R 2 are the same and are methyl or a hydrogen atom; b) either Rj or R 2 is a hydrogen atom and the other is chosen from the group consisting of methyl, iso- propyl, cyclopropylmethyl, 4-hydroxymethyl, N- methylpiperidin-4-yl, and 3-(N,N-dimethylamino)-2-methyl- prop-2-en-1-yl.
- Type I cleaved interior amine dimers are wherein R 8 is methyl, R 7 is benzyl, R 6 is 4-hydroxybenzyl, and R 3 is 4- hydroxybenzyl, and especially so wherein R 1 and R 2 are the same and are either a hydrogen atom or methyl, or one of R 1 or R 2 is a hydrogen atom and the other is methyl.
- Type I cleaved interior amine dimers are wherein R 8 is methyl, R 7 is 4- hydroxymethyl, R 6 is benzyl, and R 3 is 4-hydroxybenzyl, and especially so wherein R 1 and R 2 are the same and are either a hydrogen atom or methyl, or one of R 1 or R 2 is a hydrogen atom and the other is methyl.
- Another preferred class of compounds within the invention encompassed by Formula I is a library of an approximately equimolar mixture of two or more compounds.
- a preferred group of this library of compounds are the interior amido compounds, thus, wherein X and Y are taken together to form a carbonyl group, and especially the interior amido dimers, wherein B, AA, BB and CC are zero, except that AA can be zero or one when R 7 is a hydrogen atom and that CC can be zero or one when R 3 is a hydrogen atom.
- a preferred group of library of interior amido dimers are the resin-bound interior amido dimers, wherein R, is a solid support and R 10 is absent.
- a preferred group of these resin bound dimers are wherein R 3 and R 7 are independently chosen from the group consisting of S- or R-methyl, S- or R-benzyl, a hydrogen atom, S- or R-(but- 2-yl), S- or R-[4-(t-butoxycarbonylamino)-n-butyl], S- or R-[sec-butyl], S- or R- (methylsulfinyl) eth-1-yl, S- or R-[ 3-(guanidino)-n-propyl], S- or R- [(N-PMO-3- (guanidino)-n-propyl], S- or R-(t-butyloxy) methyl, S- or R-[2-(t-butyloxy)ethy], S-phenyl, S- or R-[2-(
- AA is one or zero when R 7 is a hydrogen atom
- CC is one or zero when R 3 is a hydrogen atom
- R 1 or R 2 are taken together with R 3 to form an S- or R- pyrrolidine or S-[4-(hydroxy)pyrrolidine].
- a more preferred group of the library of resin- bound interior amido dimers referred to hereafter as the "Type I bound amido dimers (Library)" occurs wherein R 6 and R 8 are independently methyl, ethyl, allyl, benzyl, or napth-2-ylmethyl.
- a preferred group of the Type I bound amido dimers is wherein either R 1 or R 2 is a hydrogen atom or is taken in conjunction with R 3 to form a pyrrolidine ring, and the other is C 1 to C 12 acyl, C 3 to C 10 cycloalkyl, C 3 to C 6 heterocycle, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle.
- Type I bound amido dimers (Library) is wherein either R 1 or R 2 are each a hydrogen atom, or one of R 1 or R 2 is a hydrogen atom and the other is taken together with R 3 to form an S- pyrrolidine ring.
- R 9 is hydrogen and R 10 is absent.
- a preferred group of cleaved interior amido dimers are wherein R 3 and R 7 are independently chosen from the group consisting of S- or R-methyl, S- or R- benzyl, a hydrogen atom, S- or R-(but-2-yl), S- or R-[ 4- (N-methylamino)-n-butyl], S- or R-[4-(N-ethylamino) -n- butyl], S- or R-[4-(N-allylamino)-n-butyl], S- or R-[4- (N-benzylamino)-n-butyl], S- or R-[4-(N-(napth-2- ylmethylamino)-n-butyl], S- or R-[ 4-(amino)-n-butyl], S- or R-[sec-butyl], S- or R-(methylsulfinyl)
- a more preferred group of library of cleaved interior dimers hereinafter referred to as "Type I cleaved amido dimers", wherein R 6 and R 8 are independently methyl, ethyl, allyl, benzyl, or napth-2-ylmethyl.
- R 1 and R 2 are the same and are each a hydrogen atom; (2) wherein R 6 is ethyl, R 3 is benzyl and R 1 and
- R 2 are the same and are each a hydrogen atom
- R 6 is naphth-2 -ylmethyl
- R 3 is S- methyl
- R 1 and R 2 are the same and are each a hydrogen atom
- R 1 or R 2 is a hydrogen atom and the other is taken in conjunction with R 3 to form an S-pyrrolidine ring, and R 6 is napth-2-ylmethyl.
- Another preferred group of compounds within the library of an approximately equimolar mixture of two or more compounds of Formula I are the interior amine compounds, thus, wherein in the above Formula I, X and Y are the same and are each a hydrogen atom.
- a preferred library of interior amine compounds are those that are dimers, that is wherein B, AA, BB and CC are zero, except that AA can be zero or one when R 7 is a hydrogen atom and that CC can be zero or one when R 3 is a hydrogen atom.
- Preferred interior amine dimers are those that have been cleaved from the solid support, wherein R 9 is a hydrogen atom and R 10 is absent.
- a preferred group of such cleaved interior amine dimers are wherein R 3 and R 7 are independently chosen from the group consisting of S- or R-methyl, S- or R-benzyl, a hydrogen atom, S- or R- (but-2-yl), S- or R-[4-(N-methylamino)-n-butyl], S- or R- [4-(N-ethylamino)-n-butyl], S- or R-[4-(N-allylamino)-n- butyl], S- or R-[4-(N-benzylamino)-n-butyl], S- or R-[4- (N-(napth-2-ylmethylamino)-n-butyl], S- or R-[4-(amino)- n-butyl
- AA is zero or one when R 7 is a hydrogen atom
- CC is zero or one when R 3 is a hydrogen atom
- S- or R-thiomethyl or when either R 1 or R 2 are taken together with R 3 to form an S- or R- pyrrolidine or S-[4-(hydroxy)pyrrolidine].
- R fi and R 8 are independently methyl, ethyl, allyl, benzyl, or napth-2-ylmethyl.
- Another preferred group within the library of interior amine dimers are the resin-bound compounds, thus, wherein R 9 is a solid support and R 10 is absent.
- R 6 and R 8 are independently methyl, ethyl, allyl, benzyl, or napth-2-ylmethyl.
- a preferred method of effecting analgesia in a mammal occurs when a single compound that is an interior amido dimer and further wherein B, AA, BB and CC are zero, R 9 is a hydrogen atom, R 8 is napth-2-ylmethyl, R 7 is S-methyl, R 6 is ethyl, R 3 is S-benzyl, and R 1 and R 2 are each a hydrogen atom is used.
- Another preferred method of effecting analgesia in mammals utilizes a single interior amine dimer wherein further R 9 is a hydrogen atom, R 8 is benzyl, R 7 is S-methyl, R 6 is naphth-2-ylmethyl, R 3 is taken in conjunction with either R 1 or R 2 to form an S-pyrrolidine ring and the other of R 1 and R 2 a hydrogen atom.
- a preferred method of affecting a decrease in the postprandial rise in the blood glucose of a mammal occurs wherein the single compound has X and Y taken together to form a carbonyl group, B, AA, BB and CC are zero, R 9 is a hydrogen atom, R 8 is benzyl, R 6 is naphth-2-ylmethyl, R 3 is R-(N-(naphth-2-ylmethyl)indol-3- ylmethyl), R 1 and R 2 are each hydrogen, R 10 is absent, and R 7 is chosen from the group consisting of S- (4- (N- benzylamino)-n-butyl), R-(4-(N-benzylamino)-n-butyl), S-(3-guanidino)-n-propyl), and R-(3-guanidino)-n-propyl;
- Yet another aspect of the instant invention is a method of treating microbial infections in mammals, which comprises administering an effective amount of a single compound of Formula I in conjunction with a pharmaceutically-acceptable carrier.
- a preferred method of treating microbial infections in mammals occurs when wherein the single compound has X and Y taken together to form a carbonyl group, B, AA, BB and CC are zero, R 9 is a hydrogen atom, R 8 is benzyl, R 6 is naphth-2-ylmethyl, R 3 is R-(N-(naphth-2-ylmethyl)indol-3-ylmethyl), R 1 and R 2 are each hydrogen, R 10 is absent, and R 7 is chosen from the group consisting of S-(4-(N-benzylamino)-n-butyl), R- (4-(N-benzylamino)-n-butyl), S-(3-guanidino)-n-propyl), and R-(3-guanidino)-n-prop
- Another aspect of the instant invention is a method of step-wise N-alkylation of the amide bond of the N-terminal residue of a compound of the Formula (II) :
- R 11 is independently a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycleal, or a substituted C 6 to C 15 alkyl heterocycle;
- ZZ is from zero to five
- R 12 is a solid support or a group of the Formula (III):
- R 14 is a C 1 to C 18 substituent group
- W is 0 to 4.
- R 13 is independently a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle;
- R 15 is a solid support (when W is one) or a bond to the preceeding methylene group (when W is from two to four);
- YY is from zero to five
- the compound of the above formula is a) first reacted under anhydrous conditions in an inert atmosphere with an excess amount non-nucleophilic base having a pKa between about 18 to about 40; then
- LG is leaving group
- Q is a C 1 to C 18 substituent groups as defined above for Formula (I); and repeating steps a) and b) as necessary to drive the alkylation to completion; with the proviso that all previous internal backbone amide bonds have been previously alkylated with a C 1 to C 18 substituent group and, when W is from 2 to 4, all of the R 14 groups are not the same C 1 to C 18
- a preferred method of step-wise N-alkylation occurs when LG is iodo or bromo and the -CH 2 -Q moiety is methyl, ethyl, allyl, benzyl or napth-2 -ylmethyl.
- R n and R 13 are indepently chosen from the group consisting of S- or R-methyl, S- or R-benzyl, a hydrogen atom, S- or R-(but-2-yl), S- or R-[4-(t- butoxycarbonylamino)-n-butyl], S- or R-[4-(amino)-n-butyl], S- or R-[sec-butyl], S- or R-(methylsulfinyl)eth- 1-yl, S- or R-[ 3-(guanidino)-n-propyl], S- or R-[(N- PMC)-3-(guanidino)-n-propyl], S- or R-(t-butoxy)methyl, S- or R-[2-(t-butoxy)ethy], S-phenyl, S- or R-(3-(2- butoxycarbonyl)ethyl), S- or
- R 19 , R 21 and R 23 independently are a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16
- alkylaryl a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle
- R 25 is a hydrogen atom or a solid support
- R 20 , R 22 and R 24 are independently a C 1 to C16 substituent group
- AA, BB and CC are independently 0 to 5;
- B is from 0 to 3;
- X and Y are taken together to form a carbonyl group or are separate and are each a hydrogen atom;
- R 16 , R 17 and R 18 independently are a hydrogen atom, an amino protecting group, C 1 to C 12 acyl, C 3 to C 10 cycloalkyl, C 3 to C 6 heterocycle, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle;
- R 16 is optionally present as a C 1 to C 18 substituent group when R 1 and R 2 are other than a hydrogen atom or an amino protecting group;
- said library of compounds is composed of SL physically separate sublibraries; wherein SL is equal to (2B + 4);
- each sublibrary is composed of physically separate mixtures, wherein the number of said mixtures is equal to the number of different substituents incorporated at R fix , which R fix can be any one of R 19 , R 20 , R 21 , R 22 , R 23 , or R 24 in the above Formula IV;
- step (c) Dividing the mixed solid support from step (a) into approximately equal separate portions in a number equal to the number of different substituents to be incorporated at R 24 by alkylation, alkylating each physically separate solid support mixtures with one alkyl group, then mixing said resins;
- step (g) Dividing the mixture of solid support portions from either step (c), (e), or step (f) into approximately equal separate portions equal to the number of substituents to be placed at R 19 , coupling one such monomer containing a single R 19 to each physically
- step (h) Dividing the mixture of portions from step (g) into a number of approximately equal separate portions, said number equal to the number of alkyl substituents at R 22 to be utilized, alkylating each said separate portion with a single alkyl group R 22 ;
- each coupling step in the above series of steps ( (b), (d), (f) and (g) ) involves a substrate of the Formula (V) :
- SS is a solid support
- R 26 are two hydrogen atoms each bound to the nitrogen atom
- R 28 is a C 1 to C 18 substituent group
- R 27 is independently a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle;
- R VAR can be the same or different as R 27 and is chosen from the same group of substituents as R 27 ;
- DD and EE are independently 0 to 5 ;
- X and Y are either taken together to form a carbonyl oxygen
- PG is an amino protecting group other than trityl
- A is a group, when taken with the preceeding carbonyl group; that forms an active acylating agent;
- C is from 0 to 4.
- LG is a leaving group under the conditions of the alkylation
- Q is a C 1 to C 18 substituent group as defined above in Formula (I);
- FF is 0 to 5;
- R 31 is a hydrogen atom when R 29 is a trityl group or is a C 1 to C 18 substituent group;
- R 30 is independently a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle; and
- R 29 is a trityl group when R 31 is a hydrogen atom or is a group of the Formula (IX) :
- X and Y are as X and Y above;
- GG is 0 to 5;
- C is from 1 to 4;
- R 33 is independently a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle; and R 32 is a hydrogen atom if R 32 is bonded to the N- terminal amino group or otherwise it is a C 1 to C 18 substituent wherein one such C 1 to C 18 substituent differs from the other substituents; (3) Reductive alkylation of the N- terminal nitrogen group as described above in step (i) of a compound of the Formula (X) :
- HH, II, and JJ are independently 0 to 5;
- B is from 0 to 3;
- R 39 , R 41 and R 37 are the same or different and are chosen from the group consisting of independently a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle;
- R 40 and R 42 are different and are a C 1 to C 18 substituent group
- R 34 or R 35 when B is 0, is optionally one or two hydrogen atoms attached to the nitrogen atom, or is a optionally one or more, same or different groups, chosen from the group consisting of a hydrogen atom, an amino protecting group, C 1 to C 12 acyl, C 3 to C 10 cycloalkyl, C 3 to C 6 heterocycle, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, C 7 to C 16 alkylaryl, C 7 to C 16 substituted alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle;
- R 36 is a hydrogen or a bond to a R 34 or R 35 before the reduction occurs, and when B is from 1 to 3;
- R 38 is a C 1 to C l ⁇ substituent group different from at least one other R 40 , R 42 or R 38 groups;
- Yet another aspect of the invention is an iterative synthetic approach wherein the method for the iterative synthesis and screening of a library of an approximately equimolar amount of compounds of the
- R 48 is a hydrogen atom or a solid support
- R 45 and R 47 are different and are each a C 1 to C 18 substituent group; KK and LL are independently 0 to 5; R 44 and R 46 are independently chosen from the group consisting of independently a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle;
- R 43 is one or two hydrogen atoms, or groups of the formula R a, R b and R c , wherein R a and R b independently are a hydrogen atom, an amino protecting group, C 1 to C 12 acyl, C 3 to C 10 cycloalkyl, C 3 to C 6 heterocycle, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle; R c is
- R a and R b are other than a hydrogen atom or an amino
- step (m) Coupling the monomer containing the most active R 44 substituent to each of the separate portions of resin from step (1); (n) Alkylating each of the portions from step (m) with the best alkyl group at R 45 determined in step (k);
- step (q) Alkylating each separate resin portion from step (p) with an alkylating agent placing the best alkyl group at R 47 as such alkyl group was determined in step (o);
- step (r) Coupling to each separate portion of resin the monomer containing the best R 44 substituent as such substituent was determined in step (k); (s) Alkylating each separate portion of resin with a group that was the best alkyl group R 45 as such group determined in step (k);
- each of the above coupling steps (b), (h), (1), (m), (p) or (r), involves a substrate of the Formula (XII) :
- SS is a solid support
- R 49 is two hydrogen atoms
- R 51 is a C 1 to C 18 substituent group
- R 50 is independently a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle;
- R VAR2 can be the same or different as R 50 and is chosen from the same group of substituents as R 50 ;
- MM and NN are independently 0 to 5;
- X and Y are either taken together to form a carbonyl group or are separate and are each a hydrogen atom;
- PG is an amino protecting group other than trityl
- A is a group, when taken with the preceeding carbonyl group; that forms an active acylating agent; and C is 0 or 1;
- LG is a leaving group under the conditions of the alkylation;
- Q is a C 1 to C 18 substituent group;
- 00 is 0 to 5;
- X and Y are taken together to form a carbonyl group; or are separate and are each a hydrogen atom;
- R 54 is a hydrogen atom if R 52 is a trityl group or is a C 1 to C 18 substituent group;
- R 53 is independently a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle;
- R 52 is a trityl group if R 54 is a hydrogen atom or is a group of the Formula (XVI) :
- X and Y are as X and Y above;
- PP is 0 to 5;
- R 55 is a trityl group
- R 56 is independently a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle; and
- QQ and RR are independently 0 to 5;
- R 58 and R 60 are the same or different and are chosen from the group consisting of independently of a hydrogen atom, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, phenyl, substituted phenyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle;
- R 59 and R 61 are the same or different and are a C 1 to C 18 substituent group;
- R 57 is either two hydrogen atoms attached to the nitrogen atom, or is a single hydrogen atom and another group bonded to the nitrogen atom which groups is selected from the group consisting of a hydrogen atom, an amino protecting group, C 1 to C 12 acyl, C 3 to C 10
- cycloalkyl C 3 to C 6 heterocycle, C 1 to C 12 alkyl, C 1 to C 12 substituted alkyl, C 7 to C 16 alkylaryl, C 7 to C 16 substitued alkylaryl, a C 6 to C 15 alkyl heterocycle, or a substituted C 6 to C 15 alkyl heterocycle;
- substitutent at R 1 , R 2 and R 10 would be separate variables that could be synthesized and screened by the above iterative method. Due to their location in the molecule, these substituents, if present, would be screened to find the best substituent before the N-terminal monomer and the attendant N-alkyl group could be determined.
- variable substituents and the optimum substituents at the remaining positions could be determined by the positional scanning approach.
- the N-terminal nitrogen can be reductively alkylated and quarternized and the
- R 4 and R 5 do not have to be the same as the other R 4 and R 5 groups present in the molecule. Cleavage from the solid support led to peptidomimetics of the Formula I (wherein R 9 is hydrogen, each having diversity positions at the amino acid side chain positions (R 3 , R 5 and R 7 ), at the amide alkyl groups (R 4 , R 6 , R 8 ), and at the N-terminal groups (R 1 , R 2 and R 10 ).
- lithium t-butoxide was found to be more effective for the successive formation of the amide anions.
- Reaction "i” represents the coupling of the C-terminus of peptide-like residue to an amino-derivatized solid support.
- an peptide-like residue with amino terminus protected by a base or weak-acid labile protecting group, such as Fmoc is converted to a good acylating agent in situ using known reagents and conditions.
- reagents include carbodiimide reagents (eg. N,N'-dicyclohexylcarbodiimide (DCC) and N,N'-diiso-propylcarbodiimide in conjunction with 1-hydroxybenzotriazole) in an aprotic, polar solvent such as DMF.
- LG is a good leaving group under the S n 2
- reaction labelled “iv” in Scheme 1 is a repeat of the selective N- alkylation procedure, including the preliminary N- protection steps, of reaction "ii”.
- Reaction "v” shows the removal of the trityl group from the amino-terminus of the bound residue as before followed by the removal of the selectively-N-alkylated molecule from the amino- derivatized residue with hydrogen fluoride.
- the free amino terminus can be acylated with a C 1 to C 12 acyl group, using well-known conditions as described in Staftková, M., et al., Drug Development Research, 33:146 (1994) (herein incorporated by reference). It is preferable, however, to add the R 10 (alkyl) substituent before reductive alkylation. Such an alkylation proceeds under the same alkylation conditions used for the R 4 , R 6 , and R 8 groups. Finally, while still resin-bound, or after cleavage from the resin, the interior amide groups can be reduced (i.e.
- X and Y taken together form a carbonyl group to X and Y are each a hydrogen atom.
- Such a reduction is known in the art (see, for instance, Dooley, C.T., et al., Analgesia, INRC Proceedings, 1:400 (1995)).
- R 9 is a hydrogen atom or a solid (resin)
- mild, soluble hydrogenation catalysts such as a boric acid/trimethylborate/borane-tetrahydrofuran combination can be used.
- Trt-O-Leu-MBHA resin p-methylbenzhydrylamine
- Methyl iodide, allyl bromide and benzyl bromide were used initially as alkylating agents.
- the individual crude alkylated products were analyzed by RP-HPLC and matrix assisted laser desorption ionization-mass spectroscopy (MALDI-MS) to determine their purity and identity.
- MALDI-MS matrix assisted laser desorption ionization-mass spectroscopy
- the amide alkyl groups in the second X (R 8 ) and second 0 positions (R 6 ) were: methyl, ethyl, allyl, benzyl or naphthylmethyl.
- combinatorial library consists of 250 mixtures (50 amino acids x 5 alkyl groups), each of which is composed of 230 compounds (46 amino acids x 5 alkyl groups), and was prepared applying the divide, couple and recombine process, also independently reported as the "mixing and portioning” and "split synthesis” approaches (Lam, K.S., et al., Nature, 354:82 (1991); Furka, A., et al., Int. J.
- control resins were prepared having the formula Trt-Phe-Leu-NHMe-MBHA and Trt-Ala-Trp-NHMe-MBHA. These control resins were permethylated at the first amide position to determine the completeness of the second alkylation. Following trityl removal, starting material was not detected by RP-HPLC or MALDI-MS for any of the five crude alkylation control products.
- the highly acid labile amide linkage between the peptidomimetic and the MBHA resin linker does not permit the acid labile side chain protecting groups to be removed prior to final cleavage from the resin.
- the mixtures were cleaved from the resin under standard high hydrogen fluoride cleavage conditions (Houghten, R.A., et al., Int. J. Pept. Protein Res., 27:673 (1986)) and obtained as lyophilized powders following extraction with 50% aqueous
- the nonsupport-bound library mixtures were screened in solution in radio-receptor, antimicrobial and enzyme inhibition assays. Deconvolution of the highly active mixtures was carried out by both iterative and positional scanning methods. The iterative method is set forth in Dooley, et al., Science, 266:2019-2022 (1994) and the positional scanning method is set forth in U.S. Patent Application Serial No. 07/943,709, herein
- the instant invention is directed to screening sublibraries of the selectively N-alkylated componds of Formula I wherein each sublibrary has an R group defined, and all other R groups are synthesized with the desired subtstituents, and defining each single variable in a similar grouping of sublibrairies and screening for biological activity, until all such
- variable positions have been defined and screened for the desired activity.
- One skilled in the art would realize that this approach could also be applied in the situation wherein each sublibrary has to R groups defined, using a modification of the above techniques.
- the reduction of the interior amide of the compounds of Formula I is another means for the chemical transformation of such compounds which adds stability and can enhance activity.
- a number of reagents are
- Diborane has the advantage that
- a newly synthesized compound can be purified using a method such as reverse phase high performance liquid chromatography (RP-HPLC) or other methods of separation based on the size or charge of the compound. Furthermore, the purified compound can be characterized using these and other well known methods such as amino acid analysis and mass spectrometry.
- RP-HPLC reverse phase high performance liquid chromatography
- the compounds can be assayed for receptor binding activity using the radioreceptor assay (Examples III and IV) or other assays outlined below, including the glycosidase assay (Example V).
- the compounds of the present invention can be used in in vitro assays to study the opiate receptor subtypes.
- a detectable marker such as a radioisotope
- the compounds after being labeled with a detectable marker such as a radioisotope, can be contacted with the receptor sample under conditions which specifically favor binding to a particular receptor subtype. Unbound receptor and compound can be removed, for example, by washing with a saline solution, and bound receptor can then be detected using methods well known to those skilled in the art. Therefore, the compounds of the present invention are useful in vitro for the diagnosis of relevant opioid receptor subtypes, and in particular the ⁇ type, in brain and other tissue samples.
- the compounds are also useful in vivo.
- certain of the instant compounds can be used in vivo diagnostically to localize opioid receptor subtypes.
- the compounds are also useful as drugs to treat pathologies associated with other compounds which interact with the opioid receptor system. It can be envisioned that these compounds can be used for
- morphine is a centrally acting pain killer.
- Morphine has a number of deleterious effects in the periphery which are not required for the desired analgesic effects, such as constipation and pruritus (itching). While it is known that the many peptides do not readily cross the blood- brain barrier and, therefore, elicit no central effect, the subject peptides can have value in blocking the periphery effects of morphine, such as constipation and pruritus.
- compositions comprising the compounds of Formula I in a pharmaceutically acceptable carrier.
- pharmaceutically acceptable carrier encompasses any of the standard pharmaceutical carriers, such as a phosphate buffered saline solution, water, and emulsions, such as an oil/water or water/oil emulsion, and various types of wetting agents.
- compositions will, in general, contain an effective amount of the active reagent
- compositions can take the form of tablets, pills, capsules, powders, enterically coated or other protected formulations (such as by binding on ion exchange resins or other carriers, or packaging in lipid protein vesicles or adding
- sustained release formulations e.g. ophthalmic drops
- solutions e.g. ophthalmic drops
- suspensions elixirs, aerosols, and the like.
- Water, saline, aqueous dextrose, and glycols are preferred liquid carriers, particularly (when isotonic) for
- the carrier can be selected from various oils including those of petroleum, animal, vegetable or synthetic origin, for example, peanut oil, soybean oil, mineral oil, sesame oil, and the like.
- Suitable pharmaceutical excipients include starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol, and the like.
- compositions may be subjected to conventional pharmaceutical procedures such as
- sterilization and may contain conventional pharmaceutical additives such as preservatives, stabilizing agents, wetting or emulsifying agents, salts for adjusting osmotic pressure, buffers, and the like.
- This invention provides methods of effecting treating a mammal comprising the step of administering a therapeutically effective amount of a pharmaceutical composition of this invention to a subject.
- a therapeutically effective amount is that amount necessary to alleviate the condition from which the mammal suffers.
- composition of a compound of Formula I is administered via any of the usual and acceptable methods known in the art, either singly or in combination with another compound of the present invention.
- compounds or compositions can thus be administered orally, sublingually, topically (e.g., on the skin or in the eyes), parenterally (e.g., intramuscularly,
- the administration can be conducted in single unit dosage form with continuous therapy or in single dose therapy ad libitum.
- the therapeutic methods of the present invention are practiced when the relief of symptoms is specifically required or perhaps imminently so. In another embodiment, the method is effectively practiced as continuous or prophylactic treatment.
- the particular dosage of pharmaceutical composition to be administered to the subject will depend on a variety of considerations including the nature of the disease, the severity thereof, the schedule of administration, the age and physical characteristics of the subject, and so forth. Proper dosages may be
- Topical dosages may utilize formulations containing active peptides and a liquid carrier or excipient, with multiple daily applications being appropriate.
- Fluorenylmethoxycarbonyl (Fmoc) amino acid derivatives were purchased from Calbiochem-Novabiochem Corp. (San Diego, CA, USA), Bachem Bioscience Inc.
- MBHA resin (1% divinylbenzene, 100 - 200 mesh, 0.9 mmol/g substitution), was received from
- Fmoc-His(Trt)- OH and Fmoc-D-His(Trt)-OH were also used in the N- terminal position of the library.
- the library described below was synthesized using simultaneous multiple peptide synthesis (Houghten, R.A., Proc. Natl. Acad. Sci. USA, 82:5131 (1985).
- the solid support (MBHA resin) was contained in 230
- polypropylene mesh packets 250 mg resin per packet
- the only amino acids which required repetitive couplings were Fmoc-L-Gln-OH, Fmoc-D-Gln-OH, Fmoc-L-Arg(Pmc)-OH and Fmoc-D-Lys(Boc)-OH. Removal of the Fmoc protecting group was accomplished by shaking the resin packets in 20% piperidine/DMF (1 x 3 min, 1 x 10 min; 2 L) followed by a wash cycle (5 x DMF, 2 x IPA, 3 x DCM; approximately 8 mL per packet).
- the resin of the 230 library packets was combined, mixed in DCM (2 L; 15 h shaking), and dried.
- the resin was divided into 250 polypropylene mesh packets (packet size 3 cm x 3cm; each containing 310 mg resin).
- the resin packets prepared as described above, were washed (1 x DCM; approximately 8 mL per packet), treated twice with 2% TFA in DCM (1 x 2 min, 1 x 30 min; 2 L), and washed (1 x DCM, 2 x IPA, 2 x DCM, 1 x MeOH; approximately 8 mL per packet).
- the 250 mixture resin packets were cleaved 24 at a time with hydrogen fluoride (5 mL per resin packet with 0.35 mL anisole added as scavenger) using a multiple vessel cleavage apparatus (Kornheim, W., et al., Int. J. Pept. Protein Res., 25:414 (1985)).
- the resulting mixtures were extracted by sonicating with 50% aqueous acetonitrile (3 x 5 ml).
- the resulting solutions were lyophilized and relyophilized twice more from 50% aqueous acetonitrile.
- the peptidomimetic compound was synthesized using simultaneous multiple peptide synthesis (Merrifield, R. B. J. Am. Chem. Soc. 1963, 85, 2149; Houghten, R. A. Proc. Natl. Acad. Sci. USA 1985, 82, 5131) and Fmoc strategy.
- the solid support (MBHA resin) was contained in a polypropylene mesh packet (100 mg resin per packet; packet size 3 cm x 3 cm).
- Removal of the Fmoc protecting group was accomplished by shaking the resin packet in 20% piperidine/DMF (1 x 3 min, 1 x 10 min; 2 L) followed by a wash cycle (5 x DMF, 2 x IPA, 3 x DCM).
- the resin packet (0.09 mmol of free N- ⁇ -amino groups) was shaken for 2 h in a 0.077 M solution of trityl chloride (0.45 mmol) in DCM/DMF (9:1, 5.84 mL) containing DIEA (2.61 mmol, 0.45 mL). After a short wash procedure (1 x DMF, 1 x 5% DIEA, 1 x DCM), the
- the resin packet was washed (1 x DCM; approximately 5 mL), treated twice with 2% TFA in DCM (1 x 3 min, 1 x 30 min), and washed (1 x DCM, 2 x IPA, 2 x DCM, 1 x MeOH; approximately 5 mL). 5. Coupling of the second amino acid deriva tive and second alkylation
- Resin packets containing resin bound compounds with free N-terminal amino groups were shaken in a solution of methanol (MeOH) 20%/dichloromethane (DCM) 79%/acetic acid 1% (for one resin packet containing 0.05 mmol amine 4 ml of solvent were used - enugh to cover the resin packet) and 2 - 10 equivalents of the aldehyde or ketone (depending on their reactivity). After 20 minutes, 2-10 equivalents of a 1 M solution of sodium cyanoborohydride in N,N-dimethylformamide (DMF) were added and the reaction mixture shaken for 60 min. The resin packets were washed using the following washing sequence: 5 x DMF, 1 x DCM, 1 x MeOH (for one resin packet of the size mentioned above approximately 5 ml of solvent for each step). The completeness of the
- reaction mixture was shaken for 60 min.
- the resin packet was washed using the following washing sequence: 5 x DMF, 1 x DCM, 1 x MeOH, approximately 5 ml of solvent for each step.
- the completeness of the formation of the secondary amines was tested using the Kaiser test [if necessary the reaction can be repeated, also by using a different solvent system like DMF containing 1% acetic acid].
- Reduction can either be performed on solid support [procedure A] or in solution [procedure B].
- the resin packet was washed with DMF (2 x 1 min x 5ml), DCM (2 x 1 min x 5 ml), MeOH (1 x 1 min x 5 ml), DMF (2 x 1 min x 5ml), DCM (2 x 1 min x 5 ml) and MeOH (1 x 1 min x 5 ml).
- the tubes were cooled to room temperature and 5ml methanol were added dropwise to remove excess reducing agent. Excess solvent was removed by immersion of the tube in a 55°C water bath under a constant nitrogen flow (10-15 psi). The compound subsequently underwent
- the compound was cleaved from the resin with hydrogen fluoride (5 mL per resin packet with 0.35 mL anisole added as scavenger) using a multiple vessel cleavage apparatus (Houghten, R. A., et al., Int. J.
- This example describes the identification of individual compounds, either contained within a synthetic combinatorial library mixture or prepared separately, as inhibitors of the ⁇ -selective opioid peptide [ 3 H]-[D-Ala 2 , MePhe 4 , Gly-Ol 5 enkephalin ([ 3 H]-DAMGO). Individual peptides were identified as capable of inhibiting [ 3 H] - DAMGO by a radioreceptor assay. As detailed below, the compound libraries of the instant invention were screened at a single
- Protein content of the crude homogenates ranged from
- polypropylene tubes Each tube contained 0.5 ml of membrane suspension, 3 nM of the ⁇ -selective opioid peptide [ 3 H]-DAMGO (specific activity 36 Ci/mmol), 0.08 mg/ml compound mixture and Tris-HCl buffer in a total volume of 0.65 ml. Assay tubes were incubated for 60 min at 25°C. The reaction was terminated by filtration through GF-B filters (Wallac, Inc., Gaithersburg, MD). The filters were subsequently washed with 6 ml Tris-HCl buffer at 4°C. Bound radioactivity was counted on a Beta-plate Liquid Scintillation Counter (Life Technologies, Gaithersburg, MD) and expressed in counts per minute (cpm).
- Inter- and intra-assay variation standard curves were determined by incubation of [ 3 H] - DAMGO in the presence of 0.13-3900 nM of unlabeled DAMGO.
- Competitive inhibition assays were performed as above using serial dilutions of the peptide mixtures. IC 50 values were then calculated using the software GRAPHPAD (ISI, San Diego, CA). IC 50 values of less than 1000 nM are indicative of highly active opioid compounds which bind to the ⁇ receptor, with particularly active
- Assays demonstrating selective inhibition of binding to kappa oppiate receptors for K receptors were carried out using [ 3 H]-U69,593 (3 nM, specific activity 62 Ci/mmol) as the radioligand and tissue homogenates prepared from guinea pig brains (cortex and cerebellum) using Tris buffer containing 100 ⁇ M PMSF, 5 mM MgCl 2 and 1 mg/ml BSA, pH 7.4. Sample tubes were incubated for 2.5 hr. Standard curves were prepared using 0.05-6300 nM naloxone.
- ⁇ -Glucosidase Inhibitor ⁇ -Glucosidases are not only essential to carbohydrate metabolism, but also vital for the
- Inhibitors of these enzymes are therefore of high therapeutic potential.
- ⁇ -glucosidase inhibitors are potent oral anti-diabetics
- the most active inhibitors are compounds of Formula I wherein X and Y are taken together to form a carbonyl group, B is zero, AA, BB, and CC are zero except were noted, R 9 is a hydrogen atom, R 8 is benzyl, R 6 is naphth-2-ylmethyl, R 3 is S- (N- (naphth-2-ylmethyl)indol-3-ylmethyl), R 1 and R 2 are each hydrogen, R 10 is absent, and R 7 is as set forth in the following Table 3 :
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Abstract
Description
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Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP97908448A EP0890101A4 (en) | 1996-03-05 | 1997-03-05 | Selectively n-alkylated peptidomimetic combinatorial libraries and compounds therein |
AU20405/97A AU720632B2 (en) | 1996-03-05 | 1997-03-05 | Selectively N-alkylated peptidomimetic combinatorial libraries and compounds therein |
JP09531626A JP2001502293A (en) | 1996-03-05 | 1997-03-05 | Selective N-alkylated peptidomimetic combinatorial libraries and compounds therein |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US61139096A | 1996-03-05 | 1996-03-05 | |
US08/611,390 | 1996-03-05 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1997033174A1 true WO1997033174A1 (en) | 1997-09-12 |
Family
ID=24448833
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/IB1997/000349 WO1997033174A1 (en) | 1996-03-05 | 1997-03-05 | Selectively n-alkylated peptidomimetic combinatorial libraries and compounds therein |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP0890101A4 (en) |
JP (1) | JP2001502293A (en) |
AU (1) | AU720632B2 (en) |
CA (1) | CA2248078A1 (en) |
WO (1) | WO1997033174A1 (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2001038397A1 (en) * | 1999-11-22 | 2001-05-31 | Abbott Laboratories | N-alkylated peptides having antiangiogenic activity |
WO2001055091A1 (en) * | 2000-01-25 | 2001-08-02 | The Procter & Gamble Company | Solid-supported process for making n-substituted peptide mimetic compounds |
US6777535B1 (en) | 1999-11-22 | 2004-08-17 | Abbott Laboratories | N-alkylated peptides having antiangiogenic activity |
JP2010511637A (en) * | 2006-12-06 | 2010-04-15 | 南▲開▼大学 | Polypeptide polymer and synthesis method thereof |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
IE66205B1 (en) * | 1990-06-14 | 1995-12-13 | Paul A Bartlett | Polypeptide analogs |
US5480971A (en) * | 1993-06-17 | 1996-01-02 | Houghten Pharmaceuticals, Inc. | Peralkylated oligopeptide mixtures |
-
1997
- 1997-03-05 EP EP97908448A patent/EP0890101A4/en not_active Withdrawn
- 1997-03-05 JP JP09531626A patent/JP2001502293A/en active Pending
- 1997-03-05 WO PCT/IB1997/000349 patent/WO1997033174A1/en not_active Application Discontinuation
- 1997-03-05 CA CA002248078A patent/CA2248078A1/en not_active Abandoned
- 1997-03-05 AU AU20405/97A patent/AU720632B2/en not_active Ceased
Non-Patent Citations (4)
Title |
---|
METHODS IN MOLECULAR AND CELLULAR BIOLOGY, 1996, Vol. 6, DORNER B. et al., "Generation of Peralkylated Peptidomimetic Combinatorial Libraries", pages 17-22. * |
PROC. EUR. PEPT. SYMP., 23, 1994, HOUGHTEN R.A. et al., "Libraries from Libraries: The Generation of Peptidomimetic Combinatorial Diversities", pages 459-460. * |
PROC. NATL. ACAD. SCI. U.S.A., November 1994, Vol. 91, No. 23, OSTRESH J.M. et al., "Libraries from Libraries: Chemical Transformation of Combinatorial Libraries to Extend the Range and Repertoire of Chemical Diversity", pages 11138-11142. * |
See also references of EP0890101A4 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2001038397A1 (en) * | 1999-11-22 | 2001-05-31 | Abbott Laboratories | N-alkylated peptides having antiangiogenic activity |
US6777535B1 (en) | 1999-11-22 | 2004-08-17 | Abbott Laboratories | N-alkylated peptides having antiangiogenic activity |
WO2001055091A1 (en) * | 2000-01-25 | 2001-08-02 | The Procter & Gamble Company | Solid-supported process for making n-substituted peptide mimetic compounds |
JP2010511637A (en) * | 2006-12-06 | 2010-04-15 | 南▲開▼大学 | Polypeptide polymer and synthesis method thereof |
Also Published As
Publication number | Publication date |
---|---|
JP2001502293A (en) | 2001-02-20 |
EP0890101A4 (en) | 1999-12-08 |
EP0890101A1 (en) | 1999-01-13 |
AU720632B2 (en) | 2000-06-08 |
CA2248078A1 (en) | 1997-09-12 |
AU2040597A (en) | 1997-09-22 |
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