EP0649443A1 - Matieres derivees d'oxazolone - Google Patents

Matieres derivees d'oxazolone

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Publication number
EP0649443A1
EP0649443A1 EP93916883A EP93916883A EP0649443A1 EP 0649443 A1 EP0649443 A1 EP 0649443A1 EP 93916883 A EP93916883 A EP 93916883A EP 93916883 A EP93916883 A EP 93916883A EP 0649443 A1 EP0649443 A1 EP 0649443A1
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EP
European Patent Office
Prior art keywords
group
different
chemical bond
oxazolone
same
Prior art date
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EP93916883A
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German (de)
English (en)
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EP0649443A4 (fr
Inventor
Joseph C. Hogan, Jr.
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ARQULE PARTNERS, L.P.
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LEGOMER PARTNERS LP
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Publication of EP0649443A1 publication Critical patent/EP0649443A1/fr
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Ceased legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/34—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton containing six-membered aromatic rings
    • C07C229/36—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton containing six-membered aromatic rings with at least one amino group and one carboxyl group bound to the same carbon atom of the carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C237/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups
    • C07C237/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C237/22—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton having nitrogen atoms of amino groups bound to the carbon skeleton of the acid part, further acylated
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D263/00—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings
    • C07D263/02—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings
    • C07D263/30—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D263/34—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D263/36—One oxygen atom
    • C07D263/42—One oxygen atom attached in position 5
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/001—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof by chemical synthesis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • C07K14/4703—Inhibitors; Suppressors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/022—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -X-C(=O)-(C)n-N-C-C(=O)-Y-; X and Y being heteroatoms; n being 1 or 2
    • C07K5/0222—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -X-C(=O)-(C)n-N-C-C(=O)-Y-; X and Y being heteroatoms; n being 1 or 2 with the first amino acid being heterocyclic, e.g. Pro, Trp
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06191—Dipeptides containing heteroatoms different from O, S, or N
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/02—Linear peptides containing at least one abnormal peptide link

Definitions

  • the present invention relates to the logical development of biochemical and biopharmaceutical agents and of new materials, including fabricated materials such as fibers, beads, films and gels.
  • the invention relates to the development of molecular modules derived from oxazolone (azlactone) and related structures, and to the use of these modules in the a ⁇ se bly of molecules and fabricated materials with tailored properties, which are determined by the contributions of the individual building modules.
  • the molecular modules of the invention are preferably chiral, and can be used to synthesize new compounds and fabricated materials which are able to recognize biological receptors, enzymes, genetic materials, and other chiral molecules, and are thus of great interest in the fields of biopharmaceuticals, separation and materials science.
  • the discovery of new peptide hormones has involved work with peptides; the discovery of new therapeutic steroids has involved work with the steroid nucleus; the discovery of new surfaces to be used in the construction of computer chips or sensor ⁇ ha ⁇ involved work with inorganic materials, etc.
  • the discovery of new functional molecules being ad hoc in nature and relying predominantly on serendipity, has been an extremely time-con ⁇ uming, laborious, unpredictable, and costly enterpri ⁇ e.
  • nucleotides can form complementary base pairs so that complementary single-stranded molecules hybridize resulting in double- or triple-helical structures that appear to be involved in regulation of gene expression.
  • a biologically active molecule referred to as a ligand
  • binds with another molecule usually a macromolecule referred to as ligand-acceptor (e.g. a receptor or an enzyme) , and this binding elicits a chain of molecular events which ultimately gives rise to a physiological state, e.g. normal cell growth and differentiation, abnormal cell growth leading to carcinogenesis, blood-pressure regulation, nerve-impulse generation and propagation, etc.
  • ligand-acceptor e.g. a receptor or an enzyme
  • the binding between ligand and ligand-acceptor is geometrically characteristic and extraordinarily specific, involving appropriate three-dimensional structural arrangements and chemical interactions.
  • carbohydrates are being increasingly viewed a ⁇ the components of living sy ⁇ tem ⁇ with the enormously complex structures required for the encoding of the ma ⁇ ive amount ⁇ of information needed to orchestrate the proces ⁇ e ⁇ of life, e.g., cellular recognition, immunity, embryonic development, carcinogene ⁇ i ⁇ and cell-death.
  • Thu ⁇ wherea ⁇ two naturally occurring amino acid ⁇ can be used by nature to convey 2 fundamental molecular messages, i.e., via formation of the two pos ⁇ ible dipeptide ⁇ tructures, and four different nucleotides convey 24 molecular message ⁇
  • two different mono ⁇ accharide subunits can give rise to 11 unique di ⁇ accharide ⁇
  • four dissimilar monosaccharide ⁇ can give ri ⁇ e to up to 35,560 unique tetramer ⁇ each capable of functioning a ⁇ a fundamental molecular me ⁇ sage in a given phy ⁇ iological system.
  • the ganglioside ⁇ are example ⁇ of the ver ⁇ atility and effect with which organi ⁇ ms can use saccharide structures. These molecules are glycolipid ⁇ ( ⁇ ugar-lipid compo ⁇ ites) and as such are able to position themselves at strategic locations on the cell wall: their lipid component enables them to anchor in the hydropholic interior of the cell wall, positioning their hydrophilic component in the aqueous extracellular Jiiillieu.
  • the ganglioside ⁇ (like many other ⁇ accharide ⁇ ) have been cho ⁇ en to act a ⁇ cellular ⁇ entries: they are involved in both the inactivation of bacterial toxins and in contact inhibition , the latter being the complex and poorly understood process by which normal cells inhibit the growth of adjacent cells, a property lost in most tumor cells.
  • the ⁇ tructure of ganglio ⁇ ide GM, a potent inhibitor of the toxin ⁇ ecreted by the cholera organi ⁇ m, featuring a branched complex pentameric structure is shown below.
  • glycoproteins saliva-protein composites
  • human blood-group antigens the A, B, and O blood cla ⁇ e ⁇
  • glycosylation i.e., the covalent linking with ⁇ ugar ⁇
  • glycosylation of erythropoetin causes loss of the hormone's biological activity
  • deglycosylation of human gonadotropic hormone increa ⁇ e ⁇ receptor binding but re ⁇ ults in almost complete loss of biological activity ⁇ ee Rademacher et al., Ann. Rev. Biochem 57, 785 (1988)
  • TPA tissue plasminogen activating factor
  • the di ⁇ covery of ⁇ uch desirable ligand forms has traditionally been carried out either by random ⁇ creening of molecule ⁇ (produced through chemical synthesis or isolated from natural source ⁇ ) , or by u ⁇ ing a so-called "rational" approach involving identification of a lead- structure, usually the ⁇ tructure of the native ligand, and optimization of it ⁇ propertie ⁇ through numerou ⁇ cycle ⁇ of ⁇ tructural redesign and biological testing.
  • Mo ⁇ t lead- ⁇ tructure ⁇ which have been u ⁇ ed in "rational" drug de ⁇ ign are native polypeptide ligand ⁇ of receptor ⁇ or enzyme ⁇ .
  • the majority of polypeptide ligand ⁇ e ⁇ pecially the ⁇ mall ones, are relatively unstable in physiological fluids due to the tendency of the peptide bond to undergo facile hydrolysis in acidic media or in the presence of peptidases.
  • Such ligands are decisively inferior in a pharmacokinetic ⁇ en ⁇ e to nonpeptidic compounds, and are not favored a ⁇ drug ⁇ .
  • An additional limitation of small peptides as drugs is their low affinity for ligand acceptors. This phenomenon is in sharp contrast to the affinity demonstrated by large, folded polypeptides, e.g.
  • peptide ⁇ for specific acceptors, e.g. receptors or enzyme ⁇ , which is in the subnanomolar range.
  • peptide mimetic ⁇ which bind tightly, preferably in the nanomolar range, and can with ⁇ tand the chemical and biochemical rigors of coexistence with biological fluids.
  • De ⁇ pite numerou ⁇ incremental advance ⁇ in the art of peptido imetic de ⁇ ign no general ⁇ olution to the problem of converting a polypeptide-ligand structure to a peptidomimetic has been defined.
  • "rational" peptidomimetic design i ⁇ done on an ad hoc ba ⁇ is.
  • peptidic ligands belonging to a certain biochemical clas ⁇ have been converted by groups ⁇ of organic chemists and pharmacologist ⁇ to ⁇ pecific peptidomimetic ⁇ ; however, in the majority of ca ⁇ e ⁇ the re ⁇ ult ⁇ in one biochemical area, e.g. peptida ⁇ e inhibitor design using the enzyme ⁇ ub ⁇ trate a ⁇ a lead, cannot be tran ⁇ ferred for u ⁇ e in another area, e.g. tyro ⁇ ine-kina ⁇ e inhibitor de ⁇ ign u ⁇ ing the kina ⁇ e ⁇ ub ⁇ trate as a lead.
  • the peptidomi etics that re ⁇ ult from a peptide ⁇ tructural lead u ⁇ ing the "rational" approach co pri ⁇ e unnatural ⁇ -amino acid ⁇ .
  • Many of the ⁇ e mimetic ⁇ exhibit - ⁇ >veral of the trouble ⁇ ome feature ⁇ of native peptide ⁇ (x ⁇ h also comprise ⁇ -amino acids) and are, thus, not favt, ad for use as drugs.
  • Recently, fundamental research on the u ⁇ e of nonpeptidic scaffolds, such as ⁇ teroidal or ⁇ ugar ⁇ tructure ⁇ , to anchor ⁇ pecific receptor-binding group ⁇ in fixed geometric relation ⁇ hip ⁇ have been described (see for example Hirschmann, R.
  • V. D. Huebner and D.V. Santi utilized functionalized polystyrene beads divided into portions each of which was acylated with a desired amino acid; the bead portions were mixed together and then ⁇ plit into portion ⁇ each of which was subjected to acylation with a ⁇ econd de ⁇ irable amino acid producing dipeptide ⁇ , u ⁇ ing the technique ⁇ of ⁇ olid pha ⁇ e peptide ⁇ ynthesis.
  • u ⁇ ing thi ⁇ synthetic scheme exponentially increasing numbers of peptide ⁇ were produced in uniform amount ⁇ which were then ⁇ eparately ⁇ creened for a biological activity of intere ⁇ t.
  • Zuckerman et al. (1992, Int. J. Peptide
  • Protein Res. 91:1) also have developed similar methods for the synthesis of peptide libraries and applied these methods to the automation of a modular synthetic chemistry for the production of librarie ⁇ of N-alkyl glycine peptide derivatives, called "peptoids", which are screened for activity against a variety of biochemical targets.
  • peptoids N-alkyl glycine peptide derivatives
  • Encoded combinatorial chemical synthe ⁇ e ⁇ have been described recently (S. Brenner and R.A. Lerner, 1992, Proc. Natl. Acad. Sci. USA 89:5381).
  • Cry ⁇ tallization can be valuable a ⁇ a ⁇ eparation technique but in the majority of ca ⁇ e ⁇ , e ⁇ pecially in cases involving isolation of a biomolecule from a complex biological milieu, succe ⁇ ful separation is chromatographic. Chro atographic separations are the result of reversible differential binding of the components of a mixture a ⁇ the mixture move ⁇ on an active natural, ⁇ ynthetic, or ⁇ emi ⁇ ynthetic ⁇ urface; tight- binding component ⁇ in the moving mixture leave the ⁇ urface la ⁇ t en masse re ⁇ ulting in separation.
  • substrates or support ⁇ to be u ⁇ ed in separations ha ⁇ involved either the polymerization cro ⁇ linking of monomeric molecule ⁇ under variou ⁇ condition ⁇ to produce fabricated material ⁇ ⁇ uch a ⁇ bead ⁇ , gel ⁇ , or films, or the chemical modification of various commercially available fabricated materials, e.g., sulfonation of poly ⁇ tyrene beads, to produce the desired new materials.
  • Prior art support materials have been developed to perform specific separations or type ⁇ of ⁇ eparation ⁇ and are of limited utility. Many of these materials are incompatible with biological macromolecule ⁇ , e.g. rever ⁇ e-pha ⁇ e silica frequently used to perform high pres ⁇ ure liquid chromatography can denature hydrophobic protein ⁇ and other polypeptide ⁇ .
  • a chromatographic surface i ⁇ equipped with molecule ⁇ which bind ⁇ pecifically with a component of a complex mixture that component will be ⁇ eparated from the mixture and may subsequently be released by changing the experimental conditions, (e.g. buffers, stringency, etc.)
  • This type of separation i ⁇ appropriately called affinity chromatography and remain ⁇ an extremely effective and widely u ⁇ ed ⁇ eparation technique.
  • It i ⁇ certainly much more ⁇ elective than traditional chromatographic technique ⁇ , e.g chromatography on silica, alumina, silica or alumina coated with long-chain hydrocarbons, polysaccharide and other type ⁇ of bead ⁇ or gel ⁇ , etc. , which in order to attain their maximum ⁇ eparating efficiency need to be u ⁇ ed under condition ⁇ that are damaging to biomolecule ⁇ , e.g. condition ⁇ involving high pre ⁇ sure, use of organic solvents and other denaturing agents, etc.
  • Oxazolones, or azlactones are structure ⁇ of the general formula:
  • a i ⁇ a functional group and n is 0 or 1 and typically 1-3.
  • Oxazolones containing a five-membered ring and a ⁇ ingle ⁇ ub ⁇ tituent at position 4 are typically encountered a ⁇ tran ⁇ ient intermediates which cause problematic racemization during the chemical synthe ⁇ i ⁇ of peptide ⁇ .
  • An oxazolone can in principle contain one or two ⁇ ub ⁇ tituents at the 4-po ⁇ ition. When the ⁇ e ⁇ ubstituents are not equivalent, the carbon atom at the 4-position i ⁇ asymmetric and two non- ⁇ uperimpo ⁇ able oxazolone ⁇ tructures (azlactones) result:
  • Chiral oxazolones posses ⁇ ing a ⁇ ingle 4- ⁇ ub ⁇ tituent (also known as 5(4H) -oxazolones) , derived from (chiral) natural amino acid derivatives, including activated acylamino acyl structures, have been prepared and isolated in the pure, crystalline state (Bodansky, M. ; Klausner, Y.S.; Ondetti, M.A. in "Peptide Synthesis", Second Edition, John Wiley & Sons, New York, 1976, p. 14 and references cited therein) .
  • Racemization during peptide synthe ⁇ i ⁇ becomes very extensive when the desired peptide is produced by aminolysis of activated peptidyl carboxyl, as in the case of peptide chain extension from the amino terminus, e.g. I ⁇ VI shown below (see Atherton, E. ; Sheppard, R.C. " S olid Phase Peptide Synthesis, A Practical Approach", IRL Pres ⁇ at Oxford University Press, 1989, pages 11 and 1 2) .
  • Thi ⁇ approach involves the development of oxazolone (azlactone) derivative molecular building blocks, containing appropriate atoms and functional groups which may be chiral and which are used in a modular assembly of molecules with tailored propertie ⁇ ; each module contributing to the overall properties of the as ⁇ embled molecule.
  • the oxazolone derivative building block ⁇ of the invention can be u ⁇ ed to synthesize novel molecules designed to mimic the three-dimensional structure and function of native ligands, and/or interact with the binding sites of a native receptor.
  • This logical approach to molecular construction is applicable to the synthe ⁇ is of all types of molecule ⁇ , including but not limited to imetics of peptides, proteins, oligonucleotides, carbohydrates, lipids, polymers and to fabricated materials useful in materials science. It is analogous to the modular con ⁇ truction of a mechanical device that perfor ⁇ a ⁇ pecific operation wherein each module perform ⁇ a ⁇ pecific task contributing to the overall operation of the device.
  • the invention is ba ⁇ ed, in part, on the following insights of the discoverer.
  • All ligands share a single universal architectural feature: they consist of a scaffold structure, made e.g. of amide, carbon-carbon, or phosphodie ⁇ ter bonds which support ⁇ everal functional group ⁇ in a preci ⁇ e and relatively rigid geometric arrangement.
  • Binding modes between ligands and receptors share a single universal feature as well: they all involve attractive interactions between complementary structural elements, e.g., charge- and ⁇ - type interactions, hydrophobic and van der Waals forces, hydrogen bonds.
  • a continuum of fabricated materials exists spanning a dimen ⁇ ional range from about 100 A to 1 cm in diameter compri ⁇ ing various materials of con ⁇ truction, geometrie ⁇ , morphologies, and functions, all pos ⁇ e ⁇ ing the common feature of a functional ⁇ urface which i ⁇ pre ⁇ ented to a biologically active molecule or a mixture of molecule ⁇ to achieve recognition between the molecule (or the de ⁇ ired molecule in a mixture) and the ⁇ urface.
  • Oxazolone derivative ⁇ tructures heretofore regarded as unwanted intermediates which form during the synthe ⁇ i ⁇ of peptide ⁇ , would be ideal building block ⁇ for con ⁇ tructing backbone ⁇ or ⁇ caffold ⁇ bearing the appropriate functional group ⁇ that either mimic de ⁇ ired ligand ⁇ , and/or interact with appropriate receptor binding sites, and for carrying out the synthe ⁇ is of the various parts of the functionalized scaffold orthogonally, provided that racemization of the oxazolone structure ⁇ is prevented or controlled.
  • the invention i ⁇ also based, in part, on the further recognition that such derivatives of ozaxolone ⁇ , which do not racemize, can be used as universal building blocks for the synthe ⁇ i ⁇ of ⁇ uch novel molecule ⁇ .
  • oxazolone derivative ⁇ may be utilized in a variety of way ⁇ acro ⁇ the continuum of fabricated material ⁇ de ⁇ cribed above to produce new materials capable of specific molecular recognition.
  • These oxazolone derivatives may be chirally pure and used to synthesize molecules that mimic a number of biologically active molecules, including but not limited to peptides, proteins, oligonucleotides, polynucleotide ⁇ , carbohydrates and lipids, and a variety of other polymers a ⁇ well a ⁇ fabricated materials that are useful as new materials, including but not limited to solid supports useful in column chromatography, catalysts, solid phase immunoas ⁇ ay ⁇ , drug delivery vehicles, films, and
  • “intelligent” materials designed for use in selective separation ⁇ of variou ⁇ component ⁇ of complex mixture ⁇ .
  • Working example ⁇ de ⁇ cribing the u ⁇ e of oxazolone-derived module ⁇ in the modular a ⁇ embly of a variety of molecular ⁇ tructure ⁇ are given.
  • the molecular ⁇ tructure ⁇ include functionalized ⁇ ilica ⁇ urface ⁇ u ⁇ eful in the optical re ⁇ olution of racemic mixtures; peptide mimetics which inhibit human elasta ⁇ e, protein-kina ⁇ e, and the HIV protea ⁇ e; and polymer ⁇ formed via free- radical or condensation polymerization of oxazolone- containing monomers.
  • the oxazolone-derived molecules of interest pos ⁇ e ⁇ the de ⁇ ired stereochemistry and, when required, are obtained enantiomerically pure.
  • the synthe ⁇ i ⁇ of single molecular entities the synthe ⁇ i ⁇ of librarie ⁇ of oxazolone-derived molecule ⁇ , using the techniques described herein or modifications thereof which are well known in the art to perform combinatorial chemistry, is also within the scope of the invention.
  • the oxazolone-derived molecules possess enhanced hydrolytic and enzymatic stabilitie ⁇ , and in the case of biologically active materials, are transported to target ligand-acceptor macromolecules in vivo , without cau ⁇ ing any ⁇ erious side-effects.
  • chiral oxazolones in which the asymmetric center is a 4-disub ⁇ tituted carbon, a ⁇ well a ⁇ synthetic nonchiral oxazolone ⁇ may be ⁇ ynthe ⁇ ized readily and used as molecular modules capable of controlled reaction with a variety of other molecules to produce designed chiral recognition agents and conjugates.
  • These chiral oxazolones may also be linked together, u ⁇ ing polymerizing reaction ⁇ carried out either in a ⁇ tepwi ⁇ e or chain manner, to produce polymeric biological ligand mimic ⁇ of defined seguence and stereoche i ⁇ try.
  • 4-di ⁇ ub ⁇ tituted chiral oxazolone ⁇ are extremely useful in the asymmetric functionalization of various solid supports and biological macromolecules and in the production of various chiral polymers with useful propertie ⁇ .
  • the products of all of these reactions are surprisingly stable in diverse chemical and enzymological environments, and uniquely suitable for a variety of superior pharmaceutical and high-technological application ⁇ .
  • the 4 po ⁇ ition of the oxazolone precur ⁇ or does not need to be chiral, e.g., the construction of certain polymeric materials
  • the use of oxazolones in the construction of linkers for the joining of two or more pharmaceutically useful or, simply, biologically active ligands, etc., symmetric or nonchiral oxazolones are used in chemical synthe ⁇ e ⁇ .
  • the oxazolone-derived product doe ⁇ not need to incorporate the 4-po ⁇ ition of the oxazolone precur ⁇ or in the enantiomerically pure ⁇ tate
  • oxazolone precursors which are not enantiomerically pure may be used fqr synthese ⁇ .
  • Chiral 4,4'-disubstituted oxazolone ⁇ may be prepared from the appropriate N-acyl amino acid using any of a number of standard acylation and cyclization techniques well-known to those skilled in the art, e.g.:
  • the ⁇ e may be carried out with retention of the chirality at the 4-position to produce new oxazolones. This is shown for the Michael addition to an alkenyl oxazolone as follows:
  • X S or NR and A* is a functionalized alkyl group.
  • chiral amino acid precursors for oxazolone synthe ⁇ i ⁇ may be produced u ⁇ ing stereoselective reactions that employ chiral auxiliaries.
  • An example of such a chiral auxiliary is (5)-(-)-l-dimethoxymethyl-2- methoxymethylpyrrolidme (SMPD) (Liebig's Ann. Chem. 1668 (1983)) as shown below,
  • the desired chiral amino acid may be obtained using stereo ⁇ elective biochemical ,
  • 35 oxazolone ⁇ may be prepared from mono ⁇ ub ⁇ tituted oxazolone ⁇ by alkylation of the 4-position, a ⁇ in the following tran ⁇ formation (Svnthesi ⁇ Commun.. Sept. 1984, at 763; 23 Tetrahedron Lett. 4259 (1982)) :
  • Re ⁇ olution of racemic mixture ⁇ of oxalolones may be effected using chromatography or chiral support ⁇ under ⁇ uitable conditions which are well known in the art; using fractional crystallization of stable salt ⁇ of oxazolone ⁇ with chiral acid ⁇ ; or ⁇ imply by hydrolyzizing the racemic oxazolone to the amino acid derivative and re ⁇ olving the racemic modification using standard analytical techniques.
  • a wide variety of 4-mono ⁇ ub ⁇ tituted azlactone ⁇ may be readily prepared by reduction of the corresponding un ⁇ aturated derivative ⁇ obtained in high yield from the conden ⁇ ation reaction of aldehydes, ketones, or imines with the oxazolone formed from an N-acyl glycine (49 J. Org. Chem. 2502 (1984) ; 418 Synthesis Communication ⁇ (1984) )
  • Chiral oxazolones may be subjected to ring- opening reactions with a variety of nucleophiles producing chiral molecules as ⁇ hown below:
  • R 1 and R 2 differ from one another and taken alone each ⁇ ignifie ⁇ one of the followng: alkyl including cycloalkyl and ⁇ ub ⁇ tituted form ⁇ thereof; aryl, aralkyl, alkaryl, and ⁇ ub ⁇ tituted or heterocyclic ver ⁇ ions thereof; preferred forms of R 1 and R 2 are structure ⁇ mimicking the ⁇ ide chain ⁇ of naturally- occurring amino acid ⁇ as well as various ring structures.
  • the above ring-opening reaction can be carried out either in an organic ⁇ olvent ⁇ uch a ⁇ methylene chloride, ethyl acetate, dimethyl formamide (DMF) or in water at room or higher temperatures, in the presence or ab ⁇ ence of acid ⁇ , ⁇ uch a ⁇ carboxylic, other proton or Lewi ⁇ -acids, or bases, such as tertiary amines or hydroxides, serving a ⁇ catalysts.
  • organic ⁇ olvent ⁇ uch a ⁇ methylene chloride, ethyl acetate, dimethyl formamide (DMF) or in water at room or higher temperatures, in the presence or ab ⁇ ence of acid ⁇ , ⁇ uch a ⁇ carboxylic, other proton or Lewi ⁇ -acids, or bases, such as tertiary amines or hydroxides, serving a ⁇ catalysts.
  • ⁇ tructure BYH contains nucleophilic functional groups which may interfere with the ring-opening acylation, these groups must be temporarily protected using suitable orthogonal protection strategie ⁇ ba ⁇ ed on the many protecting group ⁇ known in the art; cf., e.g., Protective Group ⁇ in Organic Synthe ⁇ i ⁇ . 2ed. , T.W. Greene and P.G.M. Wuts, John Wiley & Sons, New York, N.Y., 1991.
  • the substituents A and B shown may be of a variety of structures and may differ markedly in their physical or functional properties, or may be the same; they may also be chiral or ⁇ ymmetric.
  • a and B are preferably ⁇ elected from: 1) an amino acid derivative of the form
  • NUCL NUCL
  • n natural and ⁇ ynthetic nucleotides
  • oligonucleotides n>25
  • CH deoxyribose
  • RNA ribose
  • Thi ⁇ would include natural physiologically active carbohydrates (glucose, galactose, etc.) including related compounds such as ⁇ ialic acids, etc.
  • 4) a naturally occurring or synthetic organic structural motif.
  • This term includes any of the well known base structures of pharmaceutical compounds including pharmacophores or metabolite ⁇ thereof. These structural motifs are generally known to have ⁇ pecific desirable binding properties to ligand acceptors of interest and would include ⁇ tructure ⁇ other than tho ⁇ e recited above in 1) , 2) and 3) .
  • a reporter element ⁇ uch a ⁇ a natural or synthetic dye or a residue capable of photographic amplification which po ⁇ e ⁇ es reactive groups which may be synthetically incorporated into the oxazolone structure or reaction scheme and may be attached through the group ⁇ without adversely interfering with the reporting functionality of the group.
  • Preferred reactive groups are amino, thio, hydroxy, carboxylic acid, acid chloride, isocyanate alkyl halide ⁇ , aryl halides and oxirane group ⁇ .
  • Suitable groups include vinyl group ⁇ , oxirane group ⁇ , carboxylic acids, acid chlorides, esters, amides, lactones and lactam ⁇ .
  • a macromolecular component such as a macromolecular surface or structures which may be attached to the oxazolone modules via the various reactive groups outlined above in a manner where the binding of the attached specie ⁇ to a ligand-receptor molecule is not adversely affected and the interactive activity of the attached functionality is determined or limited by the macromolecule.
  • the molecular weight of these macromolecules may range from about 1000 Daltons to as high as possible.
  • a and/or B may be a chemical bond to a suitable organic moiety, a hydrogen atom, an organic moiety which contains a suitable electrophilic group, such a ⁇ an aldehyde, e ⁇ ter, alkyl halide, ketone, nitrile, epoxide or the like, a ⁇ uitable nucleophilic group, such as a hydroxy1, amino, carboxylate, aminde, carbanion, urea or the like, or one of the R groups defined below.
  • a and B may join to form a ring or structure which connect ⁇ to the ends of the repeating unit of the compound defined by the preceding formula or may be separately connected to other moeities.
  • a and B are a ⁇ defined above and A and B are optionally connected to each other or to other compounds;
  • X and Y are the same or different and each represents a chemical bond or one or more atoms of carbon, nitrogen, sulfur, oxygen or combinations thereof;
  • R and R' are the same or different and each is an alkyl, cycloalkyl, aryl, aralkyl or alkaryl group or a sub ⁇ tituted or heterocyclic derivative thereof, wherein R and R' may be different in adjacent n units and have a selected stereochemical arrangement about the carbon atom to which they are attached;
  • G is a connecting group or a chemical bond which may be different in adjacent n units; and e.
  • n 1
  • X and Y are chemical bond ⁇
  • a and B are different and one i ⁇ other than a chemical bond, H or R
  • G include ⁇ a NH, OH or SH terminal group for connection to the carbonyl group and G-B i ⁇ other than an amino acid re ⁇ idue or a peptide
  • n i ⁇ 1 and X, Y, and G each i ⁇ a chemical bond
  • a and B each i ⁇ other than a chemical bond, an amino acid residue or a peptide
  • n i ⁇ 1, either X or A has to include a CO group for direct connection to the NH group.
  • composition ⁇ may be u ⁇ ed to mimic variou ⁇ compound ⁇ such as peptide ⁇ , nucleotides, carbohydrates, pharmaceutical compounds, reporter compounds, polymerizable compounds or sub ⁇ trates.
  • composition is defined by the formula: where A, B, X, Y and G are as defined above.
  • At least one of A and B repre ⁇ ents an organic or inorganic macromolecular surface functionalized with hydroxyl, sulfhydryl or amine groups.
  • preferred macromolecular surfaces include ceramics such as silica and alumina, porous or nonporou ⁇ beads, polymers such as a latex in the form of bead ⁇ , membranes, gels, macroscopic surfaces, or coated version ⁇ or composite ⁇ or hybrid ⁇ thereof.
  • group A or B in the above structure is an aminimide moiety.
  • This moiety may be introduced, for example by reacting the oxazolone with an asymmetrically sub ⁇ tituted hydrazine and alkylating the resulting hydrazide, (e.g., by reaction with an alkyl halide, or epoxide) .
  • An example of such a surface is shown below.
  • Preferred aminimide ⁇ are de ⁇ cribed in a PCT application entitled MODULAR DESIGN AND SYNTHESIS OF AMINIMIDE-BASED MOLECULES USEFUL AS MOLECULAR RECOGNITION AGENTS AND NEW POLYMERIC MATERIALS (attorney docket no.: 5925-005-228) and filed of even date herewith, the content of which i ⁇ expre ⁇ sly incorporated herein by reference thereto.
  • Another embodiment of the invention relates to an oxazolone ring having the ⁇ tructure
  • A, R, R' and Y are a ⁇ de ⁇ cribed above and q is zero or 1.
  • Y is a chemical bond [see claim 36]. This ring i ⁇ u ⁇ eful for preparing the de ⁇ ired oxazolone derivatives.
  • a further embodiment of the invention exploits the capability of oxazolones with ⁇ uitable ⁇ ub ⁇ tituent ⁇ at the 2-po ⁇ ition to act as alkylating agents.
  • Appropriate substituents include vinyl group ⁇ , which make the oxazolone a Michael acceptor, haloalkyl and alkyl sulfonate-e ⁇ ter and epoxide group ⁇ .
  • Michael addition to the dc :ble bond of a chiral 2-vinyloxazolone followed by a ring opening reaction re ⁇ ults in a chiral conjugate structure.
  • This general reaction scheme illustrated for the case of a 2-vinyl azlactone derivative, is as follows: wherein X represent ⁇ a ⁇ ulfur or nitrogen atom; Y represents a sulfur, oxygen, or nitrogen atom; and ⁇ ub ⁇ tituent ⁇ A and B, a ⁇ described above, may adopt a variety of structure ⁇ , differing markedly in their phy ⁇ ical or functional propertie ⁇ or being the ⁇ ame, may be chiral or achiral, and may be preferably selected from amino acid ⁇ , oligopeptide ⁇ , polypeptide ⁇ and protein ⁇ , nucleotide ⁇ , oligonucleotide ⁇ , ligand mimetics, carbohydrates, aminimide ⁇ , or ⁇ tructures found in therapeutic agents, metabolites, dyes, photographically active chemicals, or organic molecule ⁇ having de ⁇ ired steric, charge, hydrogen-bonding or hydrophobicity characteristics, or containing poly erizable vinyl groups.
  • the Michael reaction described above is usually carried out using stoichio etric amount ⁇ of nucleophile AXH and the oxazolone in a ⁇ uitable ⁇ olvent, ⁇ uch as toluene, ethyl acetate, dimethyl formamide, an alcohol, and the like.
  • the product of the Michael addition is preferably i ⁇ olated by evaporating the reaction ⁇ olvent in vacuo and purifying the material i ⁇ olated using a technique such a ⁇ recry ⁇ tallization or chromatography.
  • Gravity- or pre ⁇ sure-chromatography on one of a variety of ⁇ upport ⁇ , e.g., ⁇ ilica, alumina, under normal- or rever ⁇ ed-pha ⁇ e conditions, in the presence of a suitable solvent sy ⁇ tem, may be u ⁇ ed for purification.
  • nucleophiles of the form ROH tend to add primarily via the ring-opening reaction, and often require acidic catalysts (e.g., BF 3 ) ; thus, X should not be oxygen.
  • acidic catalysts e.g., BF 3
  • primary amines tend to add only via ring-opening, and X should therefore not be NH.
  • Secondary amines readily add to the double bond under appropriate reaction conditions, but many can also cause ring-opening; accordingly, X or Y can be N provided A or B are not hydrogen.
  • Nucleophiles of the form RSH will exclusively add via ring-opening if the sulfhydryl group i ⁇ ionized (i.e., if the ba ⁇ icity of the reaction mixture corre ⁇ pond ⁇ to pH ⁇ 9) ; on the other hand, such nucleophiles will exclusively add via Michael reaction under non-ionizing (i.e., neutral or acidic) conditions. During the Michael addition, it is important to limit the presence of hydroxylic species in the reaction mixture (e.g., moisture) to avoid ring-opening ⁇ ide-reaction ⁇ .
  • AXH can be a ⁇ econdary amine or thiol
  • BYH can be a primary or ⁇ econdary amine or thiol, or an alcohol.
  • A is a ⁇ ubstituent selected from the foregoing list and BXH comprise ⁇ an organic or inorganic macromolecular surface, e.g., a ceramic, a porous or non-porous bead, a polymer such as a latex in the form of a bead, a membrane, a gel or a composite, or hybrid of these; the macromolecular surface is functionalized with hydroxyl, sulfhydryl or amine groups which serve a ⁇ the nucleophiles in the ring-opening reaction.
  • an organic or inorganic macromolecular surface e.g., a ceramic, a porous or non-porous bead, a polymer such as a latex in the form of a bead, a membrane, a gel or a composite, or hybrid of these; the macromolecular surface is functionalized with hydroxyl, sulfhydryl or amine groups which serve a ⁇ the nucleophiles in
  • reaction sequence i ⁇ carried out under condition ⁇ ⁇ imilar to tho ⁇ e given for the nonpolymeric cases; purification of the final product involves techniques used in the art to purify support ⁇ and other ⁇ urfaces after derivatization, such as washing, dialysis, etc.
  • the role ⁇ of AXH and BYH are rever ⁇ ed, ⁇ o that BYH is the ⁇ ub ⁇ tituent ⁇ elected from the li ⁇ t above and AXH repre ⁇ ent ⁇ a functionalized ⁇ urface.
  • reactive group ⁇ may be introduced at the 2-po ⁇ ition of the oxazolone ring via ⁇ uitable acylation ⁇ , as shown for the specific example of a benzoyl chloride derivative:
  • ring-opening addition with BYH may be carried out and followed by reaction with an appropriate AXH group, e.g. an amine ANH 2 , to give the product shown:
  • a ⁇ uitable protecting group ⁇ hown a ⁇ Bl below, may be u ⁇ ed to block an exi ⁇ ting benzylic amino group in the oxazolone; ⁇ ub ⁇ equent to the ring-opening addition of BYH the protected group i ⁇ removed u ⁇ ing ⁇ tandard technique ⁇ 5 (e.g., if the protecting group is Boc, it is removed by using dilute TFA in CH 2 C1 2 ) , and the resulting product is reacted with an appropriate electrophile, e.g., A-CH 2 -Br, thus introducing substituent A into the molecule.
  • an appropriate electrophile e.g., A-CH 2 -Br
  • oxazolone building blocks By selecting appropriate oxazolone building blocks and catenating (linking) them in one of a variety of way ⁇ , it i ⁇ po ⁇ ible to produce polymeric functionalized ⁇ caffold ⁇ , of varying length and complexity, each of which mimicks a biologically important ligand and moreover possesses features which are desired of potent drug ⁇ , such as stability in physiological media, ⁇ uperior phar acokinetics, etc.
  • the oxazolones selected for catenation contain functional groups which, when part of the oxazolone-derived scaffold, will make specific contributions to the ligand- acceptor binding interaction, as determined by previou ⁇ structural studie ⁇ on the binding interaction.
  • a hybrid molecule may be produced which has improved stability properties.
  • These structures may be repre ⁇ ented through the general conjugate ⁇ tructure given above; A and B repre ⁇ ent the polypeptide sequences flanking the inserted oxazolone- derived unit or units.
  • the polymeric, oxazolone-derived ligand sequence ⁇ may be con ⁇ tructed in one of three way ⁇ a ⁇ outlined below.
  • the oxazolone ring is opened via nucleophilic attack by the amino group of a chiral ⁇ , ⁇ *-di ⁇ ub ⁇ tituted amino acid; the re ⁇ ulting amide may be recyclized to the oxazolone, with retention of chirality, and subjected to a further nucleophilic ring- opening reaction, producing a growing chiral polymer as shown below:
  • each member of the sub ⁇ tituent pair ⁇ R 1 and R 2 , R 3 and R 4 , and R 5 and R 6 differ ⁇ from the other and taken alone each ⁇ ignifie ⁇ alkyl, cycloalkyl, or substituted version ⁇ thereof, aryl, aralkyl or alkaryl, or substituted and heterocyclic version ⁇ thereof; the ⁇ e substituent pairs can also be joined into a carbocyclic or heterocyclic ring; preferred versions of these sub ⁇ tituent ⁇ are tho ⁇ e mimicking ⁇ ide- chain ⁇ tructure ⁇ found in naturally-occurring amino acid ⁇ ; X repre ⁇ ents an oxygen, sulfur, or nitrogen atom; and A and B are the substituents described above.
  • a ⁇ tructural ⁇ pecie ⁇ , po ⁇ ses ⁇ ing (1) a terminal - OH, -SH or -NH 2 group capable of ring-opening addition to the oxazolone and (2) another terminal group capable of reacting with the amino group of a chiral ⁇ , ⁇ ! - disubstituted amino acid, may be inserted in the polymer backbone as ⁇ hown below:
  • This process may be repeated, if desired, at each ⁇ tep in the ⁇ ynthe ⁇ i ⁇ where an oxazolone ring is produced.
  • the bifunctional species used may be the same or different in the steps of the synthesis.
  • ⁇ pecial ⁇ olvent ⁇ ⁇ uch a ⁇ dipolar aprotic ⁇ olvent ⁇ (e.g., dimethyl formamide, DMF, dimethyl sulfoxide, DMSO, N- methyl pyrolidone, etc.) and chaotropic (molecular aggregate-breaking) agents (e.g., urea) will be very useful as catenations produce progre ⁇ ively larger molecule ⁇ .
  • chaotropic agents e.g., urea
  • a chiral oxazolone derivative containing a blocked terminal amino group may be prepared from a blocked, disub ⁇ tituted dipeptide, that was prepared by standard techniques known to those ⁇ killed in the art, a ⁇ shown:
  • B i ⁇ an appropriate protecting group, ⁇ uch a ⁇ Boc (t-butoxycarbonyl) or Fmoc (fluorenylmethoxycarbonyl) .
  • Boc t-butoxycarbonyl
  • Fmoc fluorenylmethoxycarbonyl
  • Y is a linker (preferably a functionalized alkyl group) ;
  • X is a nitrogen of suitable structure; an oxygen or a sulfur atom; each member of the sub ⁇ tituent pair ⁇ R 1 and R 2 , R 3 and R 4 , R nl and R n differ ⁇ from the other and taken alone each ⁇ ignifies alkyl, cycloalkyl, or functionalized versions thereof; aryl, aralkyl or alkaryl or functionalized including heterocyclic version ⁇ thereof (preferably, these R sub ⁇ tituent ⁇ mimick the ⁇ ide-chain of naturally occurring amino acid ⁇ ) ;
  • ⁇ ub ⁇ tituent R can al ⁇ o be part of a carbocyclic or heterocyclic ring;
  • a i ⁇ a substituent as described above; and C is a sub ⁇ tituent selected from the set of structure ⁇ for A; and B, i ⁇ a blocking or protecting group.
  • a preliminary step may be carried out with a suitable amino acid derivative as shown below, prepared via ⁇ tandard ⁇ ynthe ⁇ i ⁇ .
  • the addition reaction may be combined with a ring-opening acylation to produce chiral polymeric ⁇ equence ⁇ .
  • Thi ⁇ is ⁇ hown for the ca ⁇ e of alkenyl azlactone ⁇ below.
  • A denotes a ⁇ tructure of the form de ⁇ cribed above and HNu'-Z-Nu 2 H repre ⁇ ent ⁇ a structure containing two differentially reactive nucleophilic groups, such as methylamino-ethylamine, 1- amino propane-3-thiol, and so on; groups Nu 1 , Nu 2 , Nu 3 and Nu 4 need not be identical and Z is a linker ⁇ tructure a ⁇ de ⁇ cribed above.
  • Structure HNu'-Z-Nu 2 H may contain two nucleophilic group ⁇ of differential reactivity, as ⁇ tated above, or if Nu 1 and Nu 2 are of comparable reactivity one of the nucleophilic groups is protected to prevent it from competing with the other and deprotected selectively following acylation; protecting groups commonly used in the art of peptide synthe ⁇ is (e.g., for the nucleophilic groups such as amino, hydroxyl, thio, etc.) are u ⁇ eful in the protection of one of the Nu ⁇ ubstituents of the ⁇ tructure HNu'-Z-N ⁇ H.
  • oligo er ⁇ are highly u ⁇ eful biochemically because of their structural similarity to polypeptides.
  • the substituent ⁇ R can be cho ⁇ en to tailor the ⁇ teric, charge or hydrophobicity characteri ⁇ tic ⁇ of the oligomer ⁇ uch that a ver ⁇ atile polypeptide mimetic re ⁇ ult ⁇ .
  • nucleophilic ring-opening of asymmetrically disubstituted oxazolone ⁇ may be utilized to introduce a chiral re ⁇ idue or sequence in selected positions in peptides or proteins to produce hybrid molecule ⁇ with improved hydrolytic ⁇ tability or other propertie ⁇ .
  • the oxazolone used in the above aminolysi ⁇ may contain a blocked amino terminu ⁇ which, after the aminoly ⁇ is, is deblocked and used for further elongation via acylation.
  • This synthetic variation is shown below (B, stand ⁇ for a ⁇ uitable blocking group a ⁇ de ⁇ cribed above) .
  • the polypeptide synthe ⁇ is may be continued, if desired, using standard peptide-synthe ⁇ is techniques.
  • the structure below illustrates a short polymer containing nine subunit ⁇ prepared as above and detached from the ⁇ olid pha ⁇ e ⁇ ynthesis support.
  • each of the R groups signifie ⁇ alkyl, cycloalkyl, or ⁇ ub ⁇ tituted version thereof; aryl, aralkyl, alkaryl, or sub ⁇ tituted including heterocyclic ver ⁇ ion ⁇ thereof; the R group ⁇ can al ⁇ o define a carbocyclic or heterocyclic ring; preferred ⁇ tructure ⁇ for the R groups are those mimicking the structure ⁇ of the side-chains of naturally-occurring amino acids.
  • di ⁇ ubstituted chiral azlactones may be utilized to introduce a variety of novel, unnatural residue ⁇ into peptide ⁇ or protein ⁇ u ⁇ ing the following multi ⁇ tep procedure: a. Synthesis of a peptide who ⁇ e carboxyl terminal re ⁇ idue i ⁇ chiral and disubstituted, preferably via solid phase synthe ⁇ i ⁇ :
  • each of the R group ⁇ ⁇ ignifies alkyl, cycloalkyl, aryl, aralkyl or alkaryl, or sub ⁇ tituted or ⁇ uitably heterocyclic versions thereof; the R groups may also define a carbocyclic or heterocyclic ring; preferably the R group ⁇ are ⁇ tructural mimetic ⁇ of the ⁇ ide-chain ⁇ of naturally-occurring amino acids.
  • oxazolone peptide produced in ⁇ tep (b) above may be reacted with a variety of bifunctional nucleophilic molecule ⁇ to give acylation products as shown below:
  • the above acylation product may be coupled with a peptide to produce novel chiral hybrids; two coupling routes may be used.
  • a i ⁇ a group which can be conden ⁇ ed with an amino group
  • the conden ⁇ ation reaction is used for coupling.
  • A is a carboxyl group
  • condensation with a peptide amine using DCC or similar reagent produces the desired product.
  • Reaction condition ⁇ and suitable (orthogonal) protecting groups well-known in the art, ⁇ uch as those de ⁇ cribed above, are expected to be u ⁇ eful.
  • a i ⁇ a ⁇ uitable nucleophilic group e.g., hydroxyl, amino, thio, etc.
  • reaction ⁇ are run under condition ⁇ , ⁇ imilar to tho ⁇ e de ⁇ cribed above for related peptide ⁇ ynthe ⁇ e ⁇ .
  • a great variety of molecules posses ⁇ ing nucleophilic hydroxyl, thio, amino and other groups, e.g., carbohydrates, may be conjugated with peptidic and related frameworks u ⁇ ing reaction ⁇ with ⁇ uitable oxazolone ⁇ a ⁇ outlined above.
  • re ⁇ idues may be attached to or inserted into peptide chains u ⁇ ing oxazolones with reactive groups attached at the 2-position of the ring.
  • oxazolone molecular building block ⁇ may be utilized to con ⁇ truct new macromolecular ⁇ tructures capable of recognizing specific molecules ("intelligent macromolecules") .
  • R i a structure capable of molecular recognition
  • L i a linker
  • Structure R may be a native ligand of a biological ligand-acceptor, or a mimetic thereof, such as those de ⁇ cribed above.
  • Linker L may be a chemical bond or one of the linker ⁇ tructures listed above, or a sequence of subunits such as amino acids, aminimide monomers, oxazolone- derived chains of atoms or the like.
  • Polymeric coating C may be attached to the supporting platform either via covalent bonds or "shrink wrapping," i.e., the bonding that re ⁇ ult ⁇ when a ⁇ urface i ⁇ ⁇ ubjected to coating polymerization well known to tho ⁇ e skilled in the art.
  • This coating element may be 1) a thin cros ⁇ linked polymeric film 10 - 50 A in thickne ⁇ , 2) a cro ⁇ linked polymeric layer having controlled microporo ⁇ ity and variable thickness, or 3) a controlled microporosity gel.
  • the controlled microporo ⁇ ity gel may be engineered to completely fill the porou ⁇ structure of the support platform.
  • the polymeric coatings may be constructed in a controlled way by carefully controlling a variety of reaction parameters, such as the nature and degree of coating crosslinking, polymerization initiator, ⁇ olvent, concentration of reactants, and other reaction conditions, ⁇ uch a ⁇ temperature, agitation, etc., in a manner that i ⁇ well known to tho ⁇ e ⁇ killed in the art.
  • reaction parameters such as the nature and degree of coating crosslinking, polymerization initiator, ⁇ olvent, concentration of reactants, and other reaction conditions, ⁇ uch a ⁇ temperature, agitation, etc.
  • the ⁇ upport platform P may be a pellicular material having a diameter (dp) from 100 A to 1000 ⁇ , a latex particle (dp 0.1 - 0.2 ⁇ ) , a icroporou ⁇ bead (dp 1 - 1000 ⁇ ) , a porous membrane, a gel, a fiber, or a continuous macroscopic ⁇ urface.
  • the ⁇ e may be commercially available polymeric material ⁇ , ⁇ uch a ⁇ ⁇ ilica, poly ⁇ tyrene, polyacrylate ⁇ , poly ⁇ ulfone ⁇ , agaro ⁇ e, cellulose, etc.
  • the multisubunit recognition agent ⁇ above are expected to be very u ⁇ eful in the development of targeted therapeutic ⁇ , drug delivery ⁇ y ⁇ tem ⁇ , adjuvant ⁇ , diagno ⁇ tic ⁇ , chiral ⁇ elector ⁇ , ⁇ eparation systems, and tailored catalyst ⁇ .
  • the azlactone ring-opening addition reaction discu ⁇ sed above may be used to directly produce a wide variety of chiral vinyl monomers. These may be polymerized or copolymerized to produce chiral oligomers or polymer ⁇ , and may be further crosslinked to produce chiral beads, membranes, gels, coatings or compo ⁇ ite ⁇ of the ⁇ e material ⁇ .
  • Other useful monomers which may be used to produce chiral cro ⁇ slinkable polymers, may be produced by nucleophilic opening of a chiral 2-vinyl oxazolone with a suitable amino alkene or other unsaturated nucleophile.
  • Vinyl polymerization and polymer-cro ⁇ linking technique ⁇ are well-known in the art (see, e.g., U.S. Patent No. 4,981,933) and are applicable to the above preferred proces ⁇ e ⁇ .
  • the bead- ⁇ taining technique of Lam may be u ⁇ ed.
  • the technique involve ⁇ tagging the ligand-candidate acceptor, e.g., an enzyme or cellular receptor of interest, with an enzyme (e.g., alkaline phosphata ⁇ e) whose activity can give rise to color prodution thus staining library support particle ⁇ which contain active ligands-candidates and leaving ⁇ upport particles containing inactive ligand- candidate ⁇ colorle ⁇ .
  • Stained ⁇ upport particle ⁇ are phy ⁇ ically removed from the library (e.g., using tiny forceps tht are coupled to a micromanipulator with the aid of a microscope) and used to structurally identify the biologically active ligand in the library after removel of the ligand acceptor from the complex by e.g., washing with 8M guanidine hydrochloride.
  • affinity selection techniques described by Zuckermann above may be employed.
  • An especially preferred type of combinatorial library is the encoded combinatorial library, which involves the ⁇ ynthe ⁇ i ⁇ of a unique chemical code (e.g., an oligonucleotide or peptide) , that i ⁇ readily decipherable (e.g., by ⁇ equencing using traditional analytical methods) , in parallel with the synthesi ⁇ of the ligand-candidate ⁇ of the library.
  • a unique chemical code e.g., an oligonucleotide or peptide
  • the ⁇ tructure of the code i ⁇ fully descriptive of the ⁇ tructure of the ligand and u ⁇ ed to ⁇ tructurally characterize biologically active ligand ⁇ who ⁇ e structures are difficult or impo ⁇ ible to elucidate u ⁇ ing traditional analytical method ⁇ .
  • Coding ⁇ cheme ⁇ for con ⁇ truction of combinatorial librarie ⁇ have been de ⁇ cribed recently (for example, ⁇ ee S. Brenner and R.A. Lerner, Proc. Natl. Acad. Sci. USA 89, 5381 (1992); J.M. Kerr, et al. J. Am. Chem. Soc. 115, 2529 (1993)).
  • the ⁇ e and other related ⁇ cheme ⁇ are contemplated for u ⁇ e in con ⁇ tructing encoded combinatorial librarie ⁇ of oligo er ⁇ and other complex ⁇ tructure ⁇ derived from oxazolone ⁇ .
  • a ⁇ uitable ⁇ olid phase synthesis support e.g., the chloromethyl resin of Merrifield, i ⁇ split into three equal portions.
  • Each amino acyl resin portion is treated with an acid solution such as neat trifluoroacetic acid (TFA), or preferably, a 1:1 mixture of TFA and CH 2 C1 2 , to remove the t-Bu blocking group.
  • TFA trifluoroacetic acid
  • the resulting acyl amino acid resin is treated with ethyl chloroformate as described above producing the oxazolone re ⁇ in.
  • Each of the re ⁇ in portions is coupled to a different glycine protected a ⁇ t-butyl e ⁇ ter using the conditions de ⁇ cribed above; the amide product i ⁇ 0 deprotected as described above, for each of the re ⁇ in portion ⁇ and cyclized to the oxazolone u ⁇ ing the reaction with ethyl chloroformate.
  • the sugar that results from reaction of D-glucose with benzaldehyde can be readily blocked at po ⁇ ition ⁇ 1 and 6, by ⁇ equential reactions with an alcohol in the presence of acid, and tritylation using techniques well known in the art of carbohydrate chemistry.
  • the resulting ⁇ ugar, with position 3 unblocked can be used selectively as described above to derivatize a desired oxazolone structure.
  • a ⁇ uitable oxazolone can al ⁇ o be ri ⁇ g-opened by a ⁇ ugar containing reactive amino ⁇ ubstituents, i.e., an aminosaccharide or polyaminosaccharide.
  • reactive amino ⁇ ubstituents i.e., an aminosaccharide or polyaminosaccharide.
  • reaction with muramic acid i ⁇ expected to proceed as follows .
  • nucleotide and oligonucleotide structural motifs incorporating oxazolone-derived structures are contemplated including, but not limited to, the following.
  • HPLC Analysis A solution of the diastereo ⁇ teric amides wa ⁇ prepared in methylene chloride at a concentration of 7 mg/ l. This solution was injected into a DuPont Model 830 liquid chromatograph equipped with a detector set at 254 nm using a 20 ⁇ l loop valve injection system. The sample was chromatographed on a 25 cm x 0.4 cm stainless steel HPLC column packed with 5 ⁇ Spherisorb S5W silica gel using a 98/1/1 cyclohexane/n-butanol/isopropanol mobile phase at a flow rate of 0.9 ml/miri.
  • the enantiomeric amide conjugates were then quantitated using a calibration curve generated with a series of synthetic mixtures containing varying ratios of the two pure enantiomer ⁇ .
  • the pure L-isomer was purchased from Schweizerhall Inc.
  • the pure D-isomer was prepared from the commercially available D,L-racemate obtained from MTM Re ⁇ earch Chemical ⁇ /Lancaster Synthesi ⁇ Inc. by the method of Clark, Phillips and Steer (J. Chem. Soc.. Perkins Trans. I at 475 [1976]).
  • N-Acryloyl-fS)-2-difluoromethyl phenylalanine N-Acryloyl-fS-2-difluoromethyl phenylalanine.
  • the wet cake was reslurried in 100 ml methanol and refiltered a total of four times.
  • the resulting product was dried in a vacuum oven set for 30" and 60°C to yield 4.87 g functionalized silica.
  • the bonded phase was packed into a 25 cm x 0.46 cm stainless- ⁇ teel HPLC column from methanol, and successfully used to separate a series of mandelic acid derivatives using standard conditions.
  • the silica was then wa ⁇ hed twice with 50 ml toluene, sucked dry, reslurried in 250 ml toluene, refiltered, reslurried in 250 ml methanol and refiltered a total of three time ⁇ .
  • the re ⁇ ulting methanol wet cake was dried in a vacuum oven set for 30" at 60°C to yield 196.4 g aminopropyl silica.
  • the wet cake was reslurried in 100 ml methanol and refiltered a total of four times.
  • the product wa ⁇ dried in a vacuum oven set for 30" and 60°C to give 9.72 g functionalized silica.
  • the bonded phase was packed into a 25 cm x 0.46 cm stainless-steel HPLC column from methanol and successive ⁇ fully used to separate a series of t-acceptor amine derivatives using standard conditions described in the Chromatography Catalog distributed by Regis Chemical, Morton Grove, 111. 60053 (e.g., the 3,5-dinitro benzoyl derivatives of racemic 2- amino-1-butanol + alpha methyl benzye amine) .
  • Buechner filter washed with 50 ml benzene, sucked dry, reslurried in 100 ml of methanol and refiltered a total of four time.
  • the resulting methanol wet cake was dried in a vacuum oven set for 30" at 60°C to yield 19.45 g oxazolone-functionalized silica.
  • NMR 9CDC1) chemical shifts
  • CH 2 CH - splitting pattern in 6 ppm region + integration ratios diagnostic for structure.
  • FTIR + (mull) strong azlactone CO band in
  • Buechner filter washed twice with 50 ml toluene, sucked dry, reslurried in 250 ml toluene, refiltered, reslurried in 250 ml methanol and refiltered a total of three times.
  • the resulting methanol wet cake was dried in a vacuum oven set for 30" at 60°C to yield 196.4 g of mercaptopropyl silica.
  • Chiral azlactone conjugates may similarly be produced using a variety of azlactone derivatives containing at the 2-po ⁇ ition other group ⁇ capable of undergoing addition (and sequential ring-opening) reactions.
  • Example ⁇ of the ⁇ e groups include hydroxyalkyl, haloalkyl and oxirane groups.
  • Thi ⁇ example teache ⁇ the ⁇ ynthe ⁇ is of a competitive inhibitor for human elastase based on the structure of known N-trifluoroacetyl dipeptide analide inhibitors - see, e.g., 107 Eur. J. Biochem. 423 (1980); 162 J. Mol. Biol. 645 (1982) and references cited therein.
  • N-trifluoroacetyl-fS)-2-methyl leucyl-fS) -2- ethylphenylalanyl-p-isopropylanl ide 0.135 g (0.001 mol) 4-isopropyl analine is dissolved in the minimum amount of an appropriate solvent, such as acetonitrile, and 0.384 g (0.001 mol) of 2-(N-trifluoroacetyl-(S)-2-methyl leucyl)-(S)-4-methyl-4- benzyl-5-oxazolone dissolved in the minimum amount of the same solvent is added gradually to the stirred solution with cooling.
  • an appropriate solvent such as acetonitrile
  • reaction mixture is allowed to come to room remperature and is stirred at room temperature for 36 hours.
  • the solvent is then removed in vacuo to yield the solid N-trifluoroacetyl- (S)-2-methyl-leucyl-(S)-2-ethylphenylalanyl analide, useful as a competitive inhibitor of human ela ⁇ ta ⁇ e in e ⁇ sentially quantitative yield.
  • N-trifluoroacetyl-(S)-2- methylleucyl-(S)-2-methylphenylalanine lithium salt is slurried in 50 ml of an appropriate ⁇ olvent, ⁇ uch a ⁇ dry benzene, in a three-necked round-bottomed fla ⁇ k equipped with a ⁇ tirrer, heating bath, claisen head, downward condenser, thermometer and dropping funnel.
  • the system is heated to 65°C, and 1.09 g (0.01 mol) of ethyl chloroformate dissolved in 10 ml dry benzene is added over a 10-min. period.
  • This mimetic is useful as a competitive inhibitor for protease ⁇ inhibited by pepstatin.
  • N-isovaleryl-(S)-2- methylvalyl-(3S,4S)-statyl-(S)-2-methylalanyl-(3S,4S)- statine useful as a pepstatin-mimetic competitive inhibitor for aspartyl proteases which are inhibited by pepstatin (see, 23 J. Med. Chem. 27 (1980) and references cited therein) .
  • NMR (d 6 DMSO) chemical shifts, integrations and D 2 0 exchange experiments diagnostic for structure.
  • Boc-protected (3S,4S)-statine, [(3S,4S)-4- amino-3-hydroxy-6- methylheptanoic acid] was produced from the commercially available amino acid, coupled with 2-methylalanine using standard peptide synthesis methods and converted to the lithium salt using the method described below. 18.30 g (0.05 mol) of this derivative was stirred in 150 ml dry acetonitrile at room temperature, 5.45 g (0.05 mol) of ethyl chloroformate and 7.0 ml (0.05 mol) of triethylamine were sequentially added with stirring and the mixture was stirred at room temperature until ga ⁇ evolution cea ⁇ ed (1.5 hour ⁇ ) .
  • 2-(S)-methylvaline was prepared from (S)-valine by the method described by Kolbe and Barth fLiebigs Ann. Chem. at 1668 (1983)), and was acylated with isovaleryl chloride using standard acylation methods to produce N- isovaleryl-(S)-methylvaline, this was subsequently treated with one equivalent of LiOH in ethanol, followed by removal of the solvent in vacuo to yield the N- isovaleryl-(S)-methylvaline lithium salt.
  • This example teaches the synthesis of a competitive inhibitor for the HIV protease, ba ⁇ ed on the in ⁇ ertion of a chiral azlactone re ⁇ idue into a strategically important position in the scissile position of the known substrate, Ac-Ser-Leu-Asn-Phe-Pro-Ile-Val-
  • the side- chain blocking groups are subsequently removed using standard peptide deprotection techniques to yield the product MeO-D-Ser-D-Leu-D-Asn-NH-CO-(S)-Phe-[Me]-NH-CO- CH2-CH2-L-N-Pro-L-Ile-L-Val-OMe, useful as a competitive inhibitor for the HIV protease.
  • This example teaches the synthesis of another competitive inhibitor for the HIV protease.
  • the phenyl substituent is replaced with a uracil derivative
  • A. 74.08 g (1 mol) of N-methyl urea and 216.2 g (1 mol) of diethylethoxymethylenemalonate are heated together at 122 °C for 24 hour ⁇ , followed by 170°C for 12 hours to yield the 3-methyluracil-5-carboxylic acid ethyl ester in 35% yield, following recrystallization from ethyl acetate.
  • 3-methyluracil-5-carboxylic acid was treated with HCL and CH 2 0 using standard chloromethylation conditions to yield 3-methyl-5-chloromethyluracil in 52% yield, following standard work-up and recrystallization from ethyl acetate.
  • 5-oxazolone as prepared in Example 3.3.3 above was added portionwise to a ⁇ tirred solution of 1.14 g (0.02 mol) allyl amine in 75 ml of methylene chloride cooled to 0°C with an ice bath. After 15 min. the mixture was allowed to warm to room temperature, and was then stirred at room temperature for 4 hours. The solvent was stripped under aspirator vacuum on a rotary evaporator to yield 5.7 g of crude monomer, identified by NMR and FTIR analyses. The product was recrystallized from ethyl acetate to yield pure white crystalline monomer, useful for fabricating crosslinked chiral gels, beads, membranes and composites for chiral separations.
  • N-acetylglucosamidase and related proteins of similar specificity since the carbohydrate functionality can bind to these proteins (See 350 Biochim. Biophv ⁇ . Acta.
  • the dopamine-connected catechol functionality is a photographic developer, capable of photographic amplification by means of standard techniques.
  • Lucifer Yellow CH in 0.5 ml of water is added, and the mixture is shaken at room temperature for 6 hours.
  • the solvents are removed by freeze drying to yield 130 mg of the bifunctional adduct (XI) , which is useful as a ligand for competitive evaluation of the binding affinity of competitive ligands for protein kinases and structurally similar proteins.
  • This example describes preparation of an affinity coating from compound (III) as prepared in the previou ⁇ example.
  • Epoxy Silica "Epoxy Silica" .
  • the mixture was stirred in a rotary at room temperature for 15 min and then stripped, using a bath temperature of 44 °C, to a volatiles content of 15% as measured by weight loss (from 25-200 °C with a sun gun) .
  • the silica, coated as a result of exposure to the mixture of ingredients was slurried in 50 ml isooctane containing 32.0 mg VAZ0-64 (i.e., the polymerization catalyst 2,2'-azobisisobutyronitrile dissolved in 0.5 ml toluene that had been de-aerated with nitrogen. The slurry was then thoroughly de-aerated with nitrogen and subsequently stirred at 70 °C for 2 h.
  • VAZ0-64 i.e., the polymerization catalyst 2,2'-azobisisobutyronitrile
  • the coated silica was then collected by filtration and washed three times in 50 ml methanol, and air dried. Finally, the silica was heated at 120 °C for two hours to cure the coating and yield 5.4 g of coated silica.
  • the silica contained the following attached groups:
  • a one-cm glass column was packed with the
  • the IgG was then collected and the amount measured spectrophotometrically using standard calibration curves.
  • the mea ⁇ ured capacity of the packing was 12 mg IgG per ml of column volume.
  • a suitable experimental procedure is as follows.
  • the azlactone-functional support is slurried in a suitable solvent, such as CHC1 3 , and cooled to 0 °C.
  • An amount of the bifunctional nucleophile equivalent on a molar basis to the total number of surface azlactone groups present, is dissolved in the same solvent and added with shaking.
  • the mixture is then shaken at 0 °C for 6 hours, allowed to come to room temperature, and shaken at room temperature overnight.
  • the support is collected by filtration, washed with fresh solvent, re ⁇ slurried in an appropriate solvent and one equivalent of vinylazlactone, dissolved in the same solvent, is added thereto.
  • the mixture is then shaken, heated to 70 °C and held at this temperature for 12 hours. At the end of this time, the mixture is cooled and the support collected by filtration.
  • the support is then wa ⁇ hed thoroughly with fresh solvent and dried in vacuo .
  • the functional beads prepared as above are suspended in pH 7.5 aqueous phosphate buffer.
  • the beads are concentrated by centrifugation, the supernate decanted off and the beads washed five times with pH 7.5 aqueous phosphate buffer. The beads are then loaded into a 0.46 cm inner-diameter glass column and used to purify human IgG from serum using standard affinity-purification techniques.

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Abstract

L'invention concerne la conception et la synthèse de nouveaux modules moléculaires dérivés d'oxazolone, ainsi que l'utilisation des modules dans l'élaboration de nouvelles molécules et de nouvelles matières fabriquées. Les nouvelles molécules et les nouvelles matières fabriquées constituent des agents de reconnaissance moléculaires utiles dans la conception et la synthèse de médicaments, et elles présentent des applications dans les sciences de séparation et des matières.
EP93916883A 1992-06-30 1993-06-30 Matieres derivees d'oxazolone. Ceased EP0649443A4 (fr)

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US90675692A 1992-06-30 1992-06-30
US906756 1992-06-30
US4156293A 1993-04-02 1993-04-02
US41562 1993-04-02
PCT/US1993/006240 WO1994000509A1 (fr) 1992-06-30 1993-06-30 Matieres derivees d'oxazolone

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EP0738155A4 (fr) * 1993-12-28 1997-03-19 Arqule Partners L P Structure et synthese modulaire de molecules derivees de l'oxazolone
EP0743857A4 (fr) * 1994-01-05 1998-07-15 Arqule Inc Procede de preparation de polymeres ayant des proprietes specifiques
DE19714343A1 (de) * 1997-04-08 1998-10-15 Bayer Ag Chromatographische Enantiomerentrennung von Lactonen
GB9812675D0 (en) * 1998-06-11 1998-08-12 Univ Edinburgh Peptides
WO2000064845A1 (fr) * 1999-04-22 2000-11-02 Sankyo Company, Limited Cetoamides porteurs de resine et leur procede de preparation
EP2113510A1 (fr) 2008-04-30 2009-11-04 Freie Universität Berlin Dérivés de pepstatine A
CN112898218B (zh) * 2020-01-14 2022-08-26 河南师范大学 一锅法合成含三氟甲基噁唑酮类化合物的方法
CN112830882A (zh) * 2020-12-31 2021-05-25 河南绿园药业有限公司 一种对羟基苯甘氨酸甲酯合成方法

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2391191A1 (fr) * 1977-05-16 1978-12-15 Sandoz Sa Nouveaux composes polypeptidiques, lyeptidiques, leur preparation et leur application comme medicaments
EP0008097A2 (fr) * 1978-08-07 1980-02-20 BASF Aktiengesellschaft Procédé de préparation d'indolénines
EP0105665A1 (fr) * 1982-09-27 1984-04-18 Minnesota Mining And Manufacturing Company Composés contenant le groupement fonctionnel azlactone et compositions durcissables les contenant
EP0328340A2 (fr) * 1988-02-09 1989-08-16 BAUSCH & LOMB INCORPORATED Polymères hydrophiles perméables à l'oxygène
EP0433034A2 (fr) * 1989-12-12 1991-06-19 Minnesota Mining And Manufacturing Company Méthode impliquant liaisons alpha-amidoacétyles ou béta-amidopropionyles thermosensibles
EP0449488A1 (fr) * 1990-03-28 1991-10-02 Minnesota Mining And Manufacturing Company Produits d'addition Michael des azlactones
EP0525420A1 (fr) * 1991-07-01 1993-02-03 Mitsubishi Chemical Corporation Pseudopeptides et peptides caractérisés par une partie méthyle cétone substituée à la terminaison C comme inhibiteurs de thiol protéase

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Publication number Priority date Publication date Assignee Title
US4125519A (en) * 1976-10-13 1978-11-14 Murray Goodman Polypeptides containing 3,4-dihydroxyphenylalanine
JPS60215657A (ja) * 1984-04-10 1985-10-29 Mitsui Toatsu Chem Inc Ｎ−アシルフエニルアラニン類の製造法
US4996292A (en) * 1989-06-30 1991-02-26 Fox Sidney W Self-sealing artificial skin comprising copoly-alpha-amino acid
US5039813A (en) * 1990-06-29 1991-08-13 Polaroid Corporation 2-(4-alkenylphenyl)-5-oxazolones and polymers thereof
US5219731A (en) * 1991-11-01 1993-06-15 Wisconsin Alumni Research Foundation Method for preparing optically-active amino acid derivatives

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2391191A1 (fr) * 1977-05-16 1978-12-15 Sandoz Sa Nouveaux composes polypeptidiques, lyeptidiques, leur preparation et leur application comme medicaments
EP0008097A2 (fr) * 1978-08-07 1980-02-20 BASF Aktiengesellschaft Procédé de préparation d'indolénines
EP0105665A1 (fr) * 1982-09-27 1984-04-18 Minnesota Mining And Manufacturing Company Composés contenant le groupement fonctionnel azlactone et compositions durcissables les contenant
EP0328340A2 (fr) * 1988-02-09 1989-08-16 BAUSCH & LOMB INCORPORATED Polymères hydrophiles perméables à l'oxygène
EP0433034A2 (fr) * 1989-12-12 1991-06-19 Minnesota Mining And Manufacturing Company Méthode impliquant liaisons alpha-amidoacétyles ou béta-amidopropionyles thermosensibles
EP0449488A1 (fr) * 1990-03-28 1991-10-02 Minnesota Mining And Manufacturing Company Produits d'addition Michael des azlactones
EP0525420A1 (fr) * 1991-07-01 1993-02-03 Mitsubishi Chemical Corporation Pseudopeptides et peptides caractérisés par une partie méthyle cétone substituée à la terminaison C comme inhibiteurs de thiol protéase

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of WO9400509A1 *

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CA2139350A1 (fr) 1994-01-06
AU4659193A (en) 1994-01-24
WO1994000509A1 (fr) 1994-01-06
JPH08500576A (ja) 1996-01-23
EP0649443A4 (fr) 1995-11-08

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