EP0723441A1 - Molecules contenant des aminimides et materiaux utilises comme agents de reconnaissance moleculaire - Google Patents

Molecules contenant des aminimides et materiaux utilises comme agents de reconnaissance moleculaire

Info

Publication number
EP0723441A1
EP0723441A1 EP93916884A EP93916884A EP0723441A1 EP 0723441 A1 EP0723441 A1 EP 0723441A1 EP 93916884 A EP93916884 A EP 93916884A EP 93916884 A EP93916884 A EP 93916884A EP 0723441 A1 EP0723441 A1 EP 0723441A1
Authority
EP
European Patent Office
Prior art keywords
different
group
aminimide
same
chemical bond
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP93916884A
Other languages
German (de)
English (en)
Other versions
EP0723441A4 (fr
Inventor
Joseph C. Hogan, Jr.
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Arqule Inc
Original Assignee
LEGOMER PARTNERS LP
Arqule Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by LEGOMER PARTNERS LP, Arqule Inc filed Critical LEGOMER PARTNERS LP
Publication of EP0723441A1 publication Critical patent/EP0723441A1/fr
Publication of EP0723441A4 publication Critical patent/EP0723441A4/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D489/00—Heterocyclic compounds containing 4aH-8, 9 c- Iminoethano-phenanthro [4, 5-b, c, d] furan ring systems, e.g. derivatives of [4, 5-epoxy]-morphinan of the formula:
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82B—NANOSTRUCTURES FORMED BY MANIPULATION OF INDIVIDUAL ATOMS, MOLECULES, OR LIMITED COLLECTIONS OF ATOMS OR MOLECULES AS DISCRETE UNITS; MANUFACTURE OR TREATMENT THEREOF
    • B82B3/00—Manufacture or treatment of nanostructures by manipulation of individual atoms or molecules, or limited collections of atoms or molecules as discrete units
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3202—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the carrier, support or substrate used for impregnation or coating
    • B01J20/3204—Inorganic carriers, supports or substrates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
    • B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
    • B01J20/3244—Non-macromolecular compounds
    • B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
    • B01J20/3248—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such
    • B01J20/3251—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulphur
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
    • B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
    • B01J20/3244—Non-macromolecular compounds
    • B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
    • B01J20/3248—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such
    • B01J20/3253—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such comprising a cyclic structure not containing any of the heteroatoms nitrogen, oxygen or sulfur, e.g. aromatic structures
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
    • B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
    • B01J20/3268—Macromolecular compounds
    • B01J20/3272—Polymers obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
    • B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
    • B01J20/3268—Macromolecular compounds
    • B01J20/328—Polymers on the carrier being further modified
    • B01J20/3282—Crosslinked polymers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C243/00—Compounds containing chains of nitrogen atoms singly-bound to each other, e.g. hydrazines, triazanes
    • C07C243/40—Hydrazines having nitrogen atoms of hydrazine groups being quaternised
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C271/00—Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C07C271/06—Esters of carbamic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/04—Indoles; Hydrogenated indoles
    • C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
    • C07D209/14—Radicals substituted by nitrogen atoms, not forming part of a nitro radical
    • C07D209/16—Tryptamines
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means

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. Specifically, the invention relates to the development of molecular modules based on aminimide and related structures, and to the use of these modules in the assembly of molecules and
  • 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
  • nucleotides can form complementary base pairs so that complementary single-stranded molecules hybridize resulting in double- or triple-helical
  • a biologically active molecule 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
  • oligonucleotides that can be used to block or suppress gene expression via an antisense, ribozyme or triple helix mechanism.
  • sequence of the native target DNA or RNA molecule is characterized and standard methods are used to synthesize oligonucleotides representing the complement of the desired target
  • carbohydrates increasingly are being viewed as the components of living systems with the enormously complex structures required for the encoding of the massive amounts of information needed to orchestrate the processes of life, e .g . , cellular
  • Naturally occurring amino acids can be used by nature to convey 2 fundamental molecular messages, i.e., via formation of the two possible dipeptide structures, and four different nucleotides convey 24 molecular messages, two different monosaccharide subunits can give rise to 11 unique disaccharides, and four dissimilar monosaccharides can give rise to up to 35,560 unique tetramers each capable of functioning as a fundamental discreet
  • the gangliosides are examples of the versatility and effect with which organisms can use saccharide structures. These molecules are glycolipids (sugar-lipid composites) and as such are able to position themselves at strategic locations on the cell wall:
  • ganglioside GM a potent inhibitor of the toxin secreted by the cholera organism, featuring a branched complex pentameric structure is shown below.
  • glycoproteins responsible for the human blood-group antigens (the A, B, and 0 blood classes) are shown below.
  • incompatible blood classes cause formation of aggregates, or clusters and are the cause for failed transfusions of human blood.
  • glycosylation i.e., the covalent linking with sugars.
  • deglycosylation of erythropoetin causes loss of the hormone's biological activity; deglycosylation of human gonadotropic hormone increases receptor binding but results in almost complete loss of biological activity (see Rademacher et al., Ann. Rev. Biochem 57, 785 (1988); and glycosylation of three sites in tissue plasminogen activating factor (TPA) produces a glycopolypeptide which is 30% more active than the polypeptide that has been glycosylated at two of the sites.
  • TPA tissue plasminogen activating factor
  • a currently favored strategy for development of agents which can be used to treat diseases involves the discovery of forms of ligands of biological receptors, enzymes, or related macromolecules, which mimic such ligands and either boost, i . e . , agonize, or suppress, i.e., antagonize the activity of the ligand.
  • proteins for specific acceptors, e . g . receptors or enzymes, which is in the subnanomolar range.
  • peptide mimetics which bind tightly, preferably in the nanomolar range, and can withstand the chemical and biochemical rigors of coexistence with biological fluids.
  • peptidase inhibitor design using the enzyme substrate as a lead cannot be transferred for use in another area, e . g. tyrosine-kinase inhibitor design using the kinase substrate as a lead.
  • the peptidomimetics that result from a peptide structural lead using the "rational" approach comprise unnatural ⁇ -amino acids. Many of these mimetics exhibit several of the troublesome features of native peptides (which also comprise ⁇ -amino acids) and are, thus, not favored for use as drugs.
  • nonpeptidic scaffolds such as steroidal or sugar structures, to anchor specific receptor-binding groups in fixed geometric relationships have been described (see for example Hirschmann, R. et al . , 1992 J. Am. Chem. Soc., 114:9699-9701; Hirschmann, R. et al . , 1992 J. Am. Chem. Soc. , 114:9217-9218);
  • 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 split into portions each of which was subjected to acylation with a second desirable amino acid producing dipeptides, using the techniques of solid phase peptide synthesis.
  • This synthetic scheme exponentially increasing numbers of peptides were produced in uniform amounts which were then separately screened for a
  • 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
  • Crystallization can be valuable as a separation technique but in the majority of cases, especially in cases involving isolation of a biomolecule from a complex biological milieu , successful separation is
  • Chromatographic separations are the result of reversible differential binding of the
  • a chromatographic support is equipped with molecules which bind specifically with a component of a complex mixture, that component will be separated from the mixture and may be released subsequently by changing the experimental conditions (e.g. buffers, stringency, etc.)
  • This type of separation is appropriately called affinity chromatography and remains an extremely
  • hydrocarbons, polysaccharide and other types of beads or gels which in order to attain their maximum separating efficiency need to be used under conditions that are damaging to biomolecules, e . g. conditions involving high pressure, use of organic solvents and other denaturing agents, etc.
  • the aminimide building blocks of the invention can be used 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 synthesis of all types of molecules, including but not limited to mimetics of peptides, proteins, oligonucleotides, carbohydrates, lipids, polymers and to fabricated materials useful in materials science. It is analogous to the modular construction of a mechanical device that performs a specific operation wherein each module performs a
  • the invention is based, 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 phosphodiester bonds which support several functional groups in a precise 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 retype interactions, hydrophobic and Van der Waals forces, hydrogen bonds.
  • a continuum of fabricated materials exists spanning a dimensional range from 100 A to 1 cm in diameter comprising various materials of varied
  • aminimide modules may be utilized in a variety of ways across the continuum of fabricated materials described above to produce new materials capable of specific molecular recognition.
  • aminimide building blocks may be chirally pure and can be used to synthesize molecules that mimic a number of biologically active molecules, including but not limited to peptides, proteins, oligonucleotides, polynucleotides, carbohydrates, lipids, and a variety of polymers and fabricated materials that are useful as new materials, including but not limited to solid supports useful in column chromatography, catalysts, solid phase
  • the molecular structures include functionalized silica surfaces useful in the optical resolution of racemic mixtures; peptide mimetics which inhibit human elastase, protein-kinase, and the HIV protease; polymers formed via free-radical or condensation polymerization of aminimide-containing monomers; and lipid-mimetics useful in the detection, isolation, and purification of a variety of receptors.
  • the aminimide-based molecules of interest possess the desired stereochemistry and, when required, are obtained optically pure.
  • the synthesis of libraries of aminimide-based molecules 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.
  • aminimide-containing molecules possess enhanced hydrolytic and enzymatic stabilities, and in the case of biologically active materials, are transported to target ligand-acceptor macromolecules in vivo without causing any serious side-effects. 4. DETAILED DESCRIPTION OF THE INVENTION
  • the tetrasubstituted nitrogen of the aminimide group can be asymetric rendering aminimides chiral as shown by the two enantiomers below.
  • Dilute aqueous solutions of aminimides are neutral and of very low conductivity; aminimide conjugate acids are weakly acidic, pK a ⁇ 4.5.
  • a striking property of aminimides is their hydrolytic stability, under acidic, basic, or enzymatic conditions. For example, boiling trimethyl amine benzamide in 6 N NaOH for 24 hrs leaves the aminimide unchanged. Upon thermolytic treatment, at temperatures exceeding 180°C, aminimides decompose to give isocyanates as follows.
  • the aminimide group mimics several key
  • both functional groups contain a planar carbonyl unit and a tetrahedral atom linked to the acylated nitrogen
  • aspects of charge distribution e.g. both functional groups contain a carbonyl with significant negative charge development on the oxygen.
  • aminimides are valuable building blocks for the
  • the aminimide backbone is used as a scaffold for the geometrically precise
  • aminimide units can be linked in a variety of modes, using likers of diverse structures, to produce polymers of a great variety of structures. Specific molecular forms are chosen for screening and further study using several criteria. In one instance a certain aminimide structure is chosen because it is novel and has never been tested for activity as a biopharmaceutical agent or as material for device construction. In a preferable instance an aminimide ligand is chosen because it
  • the aminimide structure is the result of assembly of molecular modules each making a specific desirable contribution to the overall properties of the aminimide-containing molecule.
  • aminimides are functional groups with unusual and very desirable physiochemical
  • This alkylation is carried out in a suitable solvent such as a hydroxylic solvent e . g. water, ethanol, isopropanol or a dipolar aprotic solvent e . g. , DMF, DMSO, acetonitrile, usually with heating.
  • a suitable solvent such as a hydroxylic solvent e . g. water, ethanol, isopropanol or a dipolar aprotic solvent e . g. , DMF, DMSO, acetonitrile, usually with heating.
  • the acyl hydrazide is produced by the reaction of a 1,1-disubstituted hydrazine with an activated acyl derivative or an isocyanate, in a suitable organic solvent, e . g. methylene chloride, toluene, ether, etc. in the presence of a base such as triethylamine to
  • Activated acyl derivatives include acid chlorides, chlorocarbonates, chlorothiocarbonates, etc.; the acyl derivative may also be replaced with a suitable
  • DCC dicyclohexylcarbodiimide
  • the desired 1,1-disubstituted hydrazines may be readily prepared in a number of ways well known in the art; one is the reaction of a secondary amine with NH 2 Cl in an inert organic solvent.
  • acyl derivatives for the acylation reaction are the same as those required for the synthesis of the
  • the required hydrazinium salts may be prepared by routine alkylation of a 1,1-disubstituted hydrazines or by treatment of a tertiary amine with a haloamine (see 78 J. Am. Chem. Soc. 1211 (1956)).
  • Hydrazinium salts being chiral at nitrogen, may be resolved, e . g . by treatment with a chiral acid followed by separation of the diastereomers (e . g. using chromatography or fractional crystallization and the resulting enantiomers used in stereoselective syntheses of aminimides.
  • the ester may be saponified efficiently using LiOH in a mixture of methanol and water, producing a useful ⁇ -hydrazinium acid after neutralization of the reaction mixture with an acid.
  • Suitably protected hydrazinium carboxylates may be used in condensation reactions to produce aminimides.
  • the hydrazinium carboxylate units may be coupled with ⁇ -amino-acids or with other nucleophiles, such as amines, thiols, alcohols, etc., using standard techniques, to produce molecules of wide utility as ligand mimetics and new materials for high technological applications.
  • the ⁇ -hydrazinium esters may in turn be produced by the alkylation of a 1,1-disubstituted hydrazine with a haloester under standard reaction conditions, such as those given above for the alkylation of hydrazones.
  • these hydrazinium esters may be produced by standard alkylation of the appropriate ⁇ -hydrazino ester.
  • the required 1, 1-disubstituted hydrazine for the above reaction may be obtained by acid or base hydrolysis of the corresponding hydrazone (see 108 J. Am. Chem. Soc. 6394 (1986)); the alkylated hydrazone is produced from the monosubstituted hydrazone by the method of Hinman and Flores (24 J. Orq. Chem. 660 (1958)).
  • the monosubstituted hydrazones required above may be obtained by reduction of the Schiff base formed from an ⁇ -keto ester and a suitable hydrazone. This reduction may also be carried out stereoselectively, if desired, using DuPHOS-Rhodium catalysis (114 J. Am. Chem. Soc. 6266 (1992); 259 Science 479 (1993)), as shown: H
  • ⁇ -halo aminimides are prepared using an ⁇ -halo acyl halide.
  • halo aminimides may be reacted with nucleophiles containing reactive hydroxyl, thio, or amino groups to give complex aminimide structures.
  • a very useful and versatile synthesis of aminimides involves the one-pot reaction of an epoxide, an asymetrically disubstituted hydrazine, and an ester in a hydroxylic solvent, usually water or an alcohol, which is allowed to proceed usually at room temperature over several hours to several days.
  • R 1 , R 2 and R 3 are selected from a set of diverse structural types (e.g. alkyl, cycloalkyl, aryl, aralkyl, alkaryl or many substituted versions thereof), and R 4 and R 5 are alkyl or cycloalkyl.
  • substituent R 4 of the ester component in the above aminimide formation contains a double bond
  • an aminimide with a terminal double bond results which may be epoxidized, e .g. using a peracid under standard reaction conditions, and the resulting epoxide used as starting material for a new aminimide formation; thus a structure containing two aminimide subunits results.
  • R 2 and R 3 are used to illustrate the manner in which the hydrazine substituents R 2 and R 3 can be varied in each polymerization step to produce oligomers or polymers of diverse structures.
  • a related aminimide polymerization sequence utilizes an ester moiety bonded directly to the epoxide group.
  • An additional related polymerization sequence involves the use of bifunctional epoxides and esters of the following form and
  • X and Y are alkyl, cycloalkyl, aryl, aralkyl or alkaryl linkers.
  • Enantiomerically-pure aminimides may be produced by acylation of chiral hydrazinium salts as shown in the example below.
  • Chirally-pure hydrazinium salts may be obtained by resolution of the racemates; resolution can be
  • enantiomerically-pure aminimides may be obtained by resolution of the racemic
  • chiral epoxides produced by chiral epoxidations such as those developed by Sharpless (Asynn. Syn., J.D. Morrison ed., Vol. 5, Ch. 7 + 8, Acad. Press, New York, N.Y., 1985), and chiral esters, produced by standard procedures, may be used to produce a wide variety of chiral aminimides.
  • Chirally-pure aminimide molecular building blocks are especially preferred since they will be used to produce a vast array of molecules useful as new materials for high technological applications and as molecular recognition agents, including biological ligand mimetics to be used as drugs, diagnostics, and separation agents.
  • 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 symmetric.
  • a and B are preferably selected from
  • oligonucleotides n>25
  • DNA deoxyribose
  • RNA ribose
  • a reporter element such as a natural or synthetic dye or a residue capable of photographic amplification which possesses reactive groups which may be synthetically incorporated into the oxazolone structure or reaction scheme and may be attached through the groups without adversely interfering with the reporting functionality of the group.
  • Preferred reactive groups are amino, thio, hydroxy, carboxylic acid, acid chloride, isocyanate alkyl halides, aryl halides and oxirane groups.
  • Suitable groups include vinyl groups, oxirane groups, carboxylic acids, acid chlorides, esters, amides, lactones and
  • a macromolecular component such as a macromolecular surface or structures which may be attached to the oxazolone modules via the
  • 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 as an aldehyde, ester, alkyl halide, ketone, nitrile, epoxide or the like, a suitable nucleophilic group, such as a hydroxyl, 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 connects to the ends of the repeating unit of the compound defined by the preceding formula or may be separately connected to other moeities.
  • composition of the invention is defined by the structure
  • a and B are as
  • 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
  • R and R' are the same or different
  • each is an alkyl, cycloalkyl, aryl, aralkyl or alkaryl group or a substituted or
  • R' may be different in adjacent n units and
  • G is a chemical bond or a connecting group that includes a terminal carbon atom for attachment to the quaternary nitrogen and G may be different in adjacent n units;
  • G is a chemical bond
  • Y includes a
  • At least one of A and B represent an organic or inorganic
  • macromolecular surfaces include ceramics such as silica and alumina, porous and nonporous beads, polymers such as a latex in the form of beads, membranes, gels,
  • This functionalized surface may be represented as follows:
  • either A, B, or both contain one or more double bonds capable of undergoing free-radical
  • A, Y, R, R 1 and G are as defined above and
  • X- is an anion, such as a halogen or tosyl anion.
  • Yet another aspect of the invention relates to a lipid mimetic composition having the structure
  • R is an alkyl, cycloalkyl, aryl, aralkyl or alkaryl group or a substituted or heterocyclic derivative thereof
  • T is a linear or branched hydrocarbon having between 12 and 20 carbon atoms some of which are optionally substituted with oxygen, nitrogen or sulfur atoms or by an aromatic ring; and provided that at least two T substituents are present in the structure of the composition.
  • R n where n is an integer will be used to designate a group from the definition of R and R 1 .
  • Another aspect of the invention relates to functionalized polymers having the structure:
  • the invention also contemplates various methods of producing an aminimide-functional support.
  • One method comprises the steps of reacting a polymer or oligomer containing pendant moieties of OH, NH or SH with a compound of the formula:
  • R 1 and R 2 each represent alkyl, cycloalkyl, aryl, aralkyl or alkaryl, and R 3 is an amino acid derivative; a nucleotide derivative; a carbohydrate derivative; an organic structural motif; a reporter element; an organic moiety containing a polymerizable group; or a macromolecular component; coating the reacted polymer or oligomer onto a support to form a film thereon; and heating the coated support to crosslink the film.
  • Another method comprises the steps of coating a mixture of multifunctional esters and multifunctional epoxides onto a support to form a film thereon; and reacting the coated support with 1,1'-dialkylhydrazine to crosslink the film.
  • a third method comprises the steps of coating a mixture of an aminimide-functional vinyl monomer, a difunctional vinyl monomer and a vinyl polymerization initiator onto a support to form a film thereon; and heating the coating support to form a crosslinked film.
  • the aminimide-functionalized support prepared according to the previous methods are another aspect of the invention.
  • aminimide building blocks possessing functional groups capable of establishing predictable binding interactions with target molecules, and using synthetic techniques such as those broadly described above to effect catenation (linking) of the building blocks, it is possible to construct sequences of aminimide subunits mimicking selected native oligomers or polymers, e . g. peptides and polypeptides, which have better stability and pharmacokinetic properties than those of the native sequencers.
  • Specific syntheses of multisubunit aminimides are outlined below.
  • bifunctional species such as bromoalkyl isocyanates, 2- bromoalkyl oxazolones, etc., may be used as acylating agents under the reaction conditions given above.
  • the experimental conditions e .g. reaction-solvent, temperature and time, and purification
  • reaction solvents such as DMF, or N-methyl pyrollidone
  • chaotropic (aggregate-breaking) agents such as urea
  • alkylating and an acylating agent to form a racemic mixture of aminimides as before the use of BrCH 2 COCl is shown below, but other bifunctional species, such as bromoalkyl isocyanates, 2-bromoalkyl oxazolones, etc. may also be used.
  • the aminimine is normally not isolated, but used directly for the following reaction.
  • Bl is an appropriate protecting group such as BOC (t-butoxy carbonyl).
  • the ⁇ -hydrazinium carboxylic acids may be obtained by treatment of the esters with LiOH in MeOH/H 2 O at room temperature, as described above, and coupled with each other using condensation reactions promoted by DCC or other agents.
  • Protecting groups used in traditional peptide synthesis are expected to be useful here as well.
  • Aminimide subunits may be introduced into any position of a polypeptide via chemical synthesis, using one of the procedures outlined above, including the techniques for dealing with problematic reactions of high molecular weight species.
  • the resulting hybrid molecules are expected to have improved properties over the native molecules; for example, the aminimide group may confer greater hydrolytic and enzymatic stability to the hybrid molecule over its native counterpart.
  • moiety B contains a functional group which can be used to link additional aminimide and natural or unnatural amino acid subunits, e . g. via acylation reactions, complex hybrid structures may be obtained using the experimental procedures outlined above.
  • Chiral (as well as achiral) vinylaminimide monomers of the general structures shown below may be readily prepared, following the procedures outlined above, and used in free-radical polymerizations, according to experimental procedures well-known in the art, to produce a vast array of novel polymeric materials.
  • Additional monomeric structures useful in preferred free radical polymerizations include those shown below; they produce polymeric chains capable of being crosslinked into more rigid structures.
  • the monomers shown below may be prepared using the synthetic procedures outlined above, and the polymerization/crosslinking reactions may be run using standard polymerization techniques. See, for example, Practical Macromolecular Organic Chemistry, Braun, Cherdron and Kern, trans, by K. Ivin, 3ed., Vol Z, Harwood Academic Publishers, New York, N.Y. 1984.
  • the monomers shown above may be polymerized with other alkenes or dienes, which are either
  • Sequential condensations of aminimide-forming molecules may be used to produce a variety of novel polymers of controlled size.
  • An example involving dimeric epoxides and esters is given below; processes involving trimeric and more complex epoxides and esters are also contemplated; and experimental conditions for running these polymerizations (including techniques for resolving experimental difficulties as product molecular weight increases) have been described above.
  • a support e . g. silica
  • a support capable of specific molecular recognition is produced; an example of such a support is given below:
  • Aminimide conjugate structures containing two long-chain alkyl groups capable of producing bilayer membrane structures are preferred embodiments of the present invention. Many uses of these amphiphilic, surface-active compounds are envisioned. They may be used to isolate and stabilize biologically-active
  • Substituents R may be chosen from a variety of structures of various sizes including structures of ligands of biological receptors or enzymes; a preferred combination of substituents involves sterically small groups for R 1 and R 2 and a group such as A or B described above for R 3 ; the long-chain alkyl groups are 4-20 carbons i-n length; group X is a linker composed of atoms chosen from the set of C, H, N, O, and S.
  • X is a linker group (e.g. CH);
  • substituents R are chosen from the group of structures A and B described above and the remaining
  • substituent(s) in preferably a sterically small group, e .g. H, or CH 3 .
  • aminimide molecular building blocks may be utilized to construct new macromolecular structures capable of recognizing specific molecules ("intelligent macromolecules").
  • the "intelligent macromolecules” may be represented by the following general formula:
  • R is a structure capable of molecular
  • L is a linker
  • P is a macromolecular structure serving as a supporting platform
  • C is a polymeric structure serving as a coating which surrounds P.
  • Structure R may be a native ligand or a biological ligand-acceptor or a mimetic thereof, such as those described above.
  • Linker L may be a chemical bond or one of the linker structures listed above, or a sequence of subunits such as amino acids, aminimide monomers, oxazolonederived chains of atoms, etc.
  • Polymeric coating C may be attached to the supporting platform either via covalent bonds or "shrink wrapping," i.e. the bonding that results when a surface is subjected to coating polymerization well known to those skilled in the art.
  • This coating element may be
  • the controlled microporosity gel may be engineered to completely fill the porous 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, solvent, concentration of reactants, and other reaction conditions, such as temperature, agitation, etc., in a manner that is well known to those skilled in the art.
  • the support 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 microporous bead (dp 1 - 1000 ⁇ ), a porous membrane, a gel, a fiber, or a continuous macroscopic surface. These may be
  • polymeric materials such as silica, polystyrene, polyacrylates, polysulfones,
  • any of the elements P, C, L, or R containing an aminimide-based structure is derived from a form of the element containing a precursor to the aminimide-based structure.
  • the multisubunit recognition agents above are expected to be very useful in the development of targeted therapeutics, drug delivery systems, adjuvants,
  • surface refers to either P, P linked to C or P linked to C and L as defined above.
  • another aspect of the invention relates to a three-dimensional crosslinked random copolymer containing, in copolymerized form about 1 to 99 parts of a free-radically polymerizable monomer containing an aminimide group; up to 98 parts of a free-radically addition-polymerizable comonomer; and about 1 to 50 parts of at least one crosslinking monomer.
  • the comonomer used in this copolymer may be water-soluble or water-insoluble, and the copolymer is fashioned into a water-insoluble bead, a water-insoluble membrane or a latex particle, or can be a swollen aqueous gel suitable for use as an electrophoresis gel.
  • This copolymer is preferably the reaction product of about 1 to 99 parts of a condensation-polymerizable monomer containing a moiety cluster
  • chromatographic support materials for chromatographic and other applications, as well as other fabricated materials may be derivatized with tailored aminimide moieties, through chemical modification, producing novel materials capable of recognizing specific molecular structures.
  • A is selected from the group consisting of amino acids, oligopeptides, polypeptides and proteins, nucleotides, oligonucleotides,
  • X and Y are chemical bonds or groups consisting of atoms selected from the set of C, H, N, O, S ;
  • R 1 and R 2 are chosen from the group of alkyl , cycloalkyl, aryl, aralkyl, alkaryl and, preferably, structures mimicking the side-chains of naturally-occurring amino acids.
  • a surface bearing ester groups can be treated with an epoxide, containing desired group B, and a disubstituted hydrazine to form an aminimide surface as follows:
  • the surface is treated with a solution containing a 10% molar excess of the epoxide (based on the calculated number of reactive ester groups of the surface), and a stoichiometric amount of the hydrazine (with respect to the amount of the epoxide) in an appropriate solvent, such as an alcohol, with shaking.
  • an appropriate solvent such as an alcohol
  • the surface is treated with a solution containing a 10% molar excess of the ester (based on the calculated number of reactive epoxide groups of the support), and a stoichiometric amount of the hydrazine (with respect to the amount of the ester used), in an appropriate solvent, such as an alcohol, with shaking.
  • an appropriate solvent such as an alcohol
  • the foregoing reaction can be modified by utilizing an ester whose substituent B contains a double bond.
  • the double bond of the ester can be epoxidized using one of a variety of reactions including the asymetric epoxidation of Sharpies (e .g. , utilizing a peracid under suitable reaction conditions well-known in the art), and the product used as the epoxide in a new repetition of the aminimide-forming reaction.
  • the overall process can be repeated to form oligomers and polymers.
  • R 1...n and R' 1...n are used to illustrate the manner in which the hydrazine substituents R 2 and R 3 are varied in each polymerization step, if desired, to produce an oligomer or polymer.
  • Y is a linker as defined above, to form desirable polymers. If an ester-functionalized surface is reacted with bifunctional esters and epoxides, the resulting surface will have the following general structure.
  • An amine-functionalized surface can be
  • reaction (a) a 10% molar excess of methyl acrylate (based on the number of reactive amino groups the surface as determined by a titration with acid) is dissolved in an appropriate solvent, such as an alcohol, and added to the surface. After addition is complete, the mixture is shaken at room temperature for 2 days .
  • an appropriate solvent such as an alcohol
  • the solvent is then removed by decantation and the surface is washed thoroughly with fresh solvent in preparation for the next step.
  • reaction (b) the stoichiometric amount of a 1:1 mixture of the hydrazine and the epoxide, is combined in an appropriate solvent, such as an alcohol, and quickly added to the solvent-wet surface from reaction (a). The mixture is shaken at room temperature for 3 days. The solvent is then removed by decantation, and the surface is washed thoroughly with fresh solvent and dried.
  • an appropriate solvent such as an alcohol
  • substituent B in the structure above represents the surface and the desired functional group bears the amine moiety.
  • substituent B in the structure above represents the surface and the desired functional group bears the amine moiety.
  • One way of obtaining such a surface is to react a silica surface with a silicic ester containing an epoxide group to produce a so-called “epoxy silica”, as shown below.
  • a surface functionalized with a carboxylic acid group can be reacted with an 1,1-dialkylhydrazine and a coupling agent, such as dicyclohexyl carbodiimide (DCC), to form a hydrazone-containing surface as shown in step
  • a coupling agent such as dicyclohexyl carbodiimide (DCC)
  • This surface can then be coupled with a desired group B bearing a substituent capable of
  • Substituent B is a surface functionalized with an alkylating agent capable of reacting with a hydrazone.
  • the surface is treated with a 10% molar excess equimolar amounts of the N,N- dimethylhydrazine and DCC in a suitable solvent, such as methylene chloride, and the mixture is shaken for 2 hours at room temperature. The slurry is then removed by decantation and the surface is washed thoroughly with fresh solvent to remove any residual precipitated
  • the surface is then treated with a stoichiometric amount of the alkylating agent in a suitable solvent, warmed to 70 °C and held at this temperature for 6 hours. The mixture is then cooled, the solvent is removed by decantation, and the surface is washed with fresh solvent and dried.
  • a surface bearing a group capable of alkylating acyl hydrazones can be functionalized to contain
  • Z and W are linkers composed of atoms selected from the set of C, N, H, O , S , and X is a suitable leaving group, such as a halogen or tosylate.
  • a hydrazone bearing a desired group B is produced by reacting the appropriate 1,1'-dialkylhydrazine with any of a variety of derivatives containing B via reactions that are well-known in the art.
  • derivatives may be acid halides, azlactones (oxazolones), isocyanates, chloroformates, or
  • chloromethyl aminimides can be prepared by known literature procedures (See, e . g. , 21 J. Polymer Sci., Polymer Chem. Ed. 1159 (1983)), or by using the techniques described above. 4.4.6.1.6 Functionalization of Oxazolone-Containing
  • Oxazolone-containing surfaces can be functionalized by first reacting them with 1,1'- dialkylhydrazine as shown in step (a) below followed by alkylation of the resulting hydrazone with an alkylating agent B-CH 2 -X as shown in step (b); reaction conditions similar to those described above are expected to be effective in carrying out these modifications.
  • R 3 and R 4 are derived from the five membered azlactone ring denoted by Az.
  • aminimidefunctionalized composite support materials by coating various soluble aminimide formulations on the surfaces of existing supports, and subsequently crosslinking the resulting coatings in place to form mechanically stable surfaces.
  • the coating may be engineered for a particular application (e.g., to take the form of a thin non-porous film or to possess localized microporosity for enhanced surface area) by judicious selection of process
  • any of the foregoing reactions can be carried out with a vinyl aminimide in contact with a selected surface, which is polymerized according to well- known techniques (see, e .g. , U.S. Patent No. 4,737,560).
  • the polymerization results in a surface coated with a polymer containing aminimide side-chains.
  • Other coating procedures employing aminimide functional groups are described below in greater detail.
  • new surfaces and other materials can be fabricated de novo from aminimide precursors bearing polymerizable groups by polymerizations and/or
  • crosslinking agents Depending upon the properties for the desired material, various combinations of monomers, crosslinkers, and ratios thereof may be employed.
  • the resultant support materials may be latex particles, porous or non-porous beads, membranes, fibers, gels, electrophoresis gels, or hybrids thereof.
  • the monomers and crosslinking agents may or may not all be aminimides.
  • Vinyl or condensation polymerizations may be advantageously employed to prepare the desired aminimide-containing materials.
  • suitable examples include styrene, vinyl acetate, and acrylic monomers.
  • compatible non-aminimide crosslinkers such as divinyl benzene, may be employed (either singly or in combination as the other such agents).
  • Condensation polymerization may be accomplished using multifunctional epoxides and multifunctional esters with the appropriate amounts of an 1,1'-dialkylhydrazine, using the reaction conditions described above.
  • Either the ester component or the epoxide component should be at least trifunctional to obtain three-dimensionally
  • crosslinked polymer structures preferably, both
  • crosslinker to total monomer content can be varied to produce a variety of product structures (e .g. , beads, fibers, membranes, gels, or hybrids of the foregoing) and to tailor the mechanical and surface properties of the final product (e.g., particle size and shape, porosity, and surface area).
  • product structures e.g. , beads, fibers, membranes, gels, or hybrids of the foregoing
  • mechanical and surface properties of the final product e.g., particle size and shape, porosity, and surface area
  • the bead-staining technique of Lam may be used.
  • the technique involves tagging the ligand-candidate acceptor (e . g. , an enzyme or cellular receptor of interest) with an enzyme (e.g., alkaline phosphatase) whose activity can give rise to color production thus staining library support particles which contain active ligand-candidates and leaving support particles containing inactive ligand- candidates colorless.
  • an enzyme e.g., alkaline phosphatase
  • Stained support particles are physically removed from the library (e.g., using tiny forceps that 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.
  • combinatorial library is the encoded combinatorial library, which involves the synthesis of a unique chemical code (e.g., an oligonucleotide or peptide), that is readily
  • decipherable e . g. , by sequencing using traditional analytical methods
  • the structure of the code is fully descriptive of the structure of the ligand and used to structurally characterize biologically active ligands whose structures are difficult or
  • a suitable solid phase synthesis support e.g., the chloromethyl resin of Merrifield is treated with 4- hydroxyl butyric acid in the presence of CsCO 3 followed by tosylation with p-toluenesulfonyl chloride, under conditions known in the art;
  • Each resin portion is coupled with a different hydrazine followed by reaction with an acid chloride to produce a resin with three linked aminimide subunits;
  • the resin portions are mixed producing a library containing 27 types of beads each bead type containing a single trimeric aminimide species for screening using the bead-stain method described above.
  • the aminimides may be detached from the support via acidolysis producing a "solution-phase" library of aminimides containing a butyrylated terminal nitrogen. (Shown in the structure below in which
  • polysaccharide structural motifs incorporating aminimide structures are contemplated including, but not limited to, the following.
  • aminimide-based structures are contemplated including, but not limited to, the following.
  • oligonucleotides the following approach is one of many that is expected to be useful.
  • oligonucleotide-derived units an approach such as the following is expected to be very useful .
  • conjugate (II) is useful as a stabilization agent for the isolation and
  • Rhodamine B (VI), prepared from rhodamine B by the standard techniques for preparing acid chlorides from carboxylic acids, are dissolved in 500 ml of a suitable solvent and are added, with stirring, over a 1-hour period to a solution of 6.01 g (0.1 mol) of 1,1-dimethylhydrazine in 100 ml of the same solvent. The temperature is kept at 10 °C. After the addition is complete, the mixture is stirred at room temperature for 12 hours, and the solvent is stripped away in vacuo to yield the Rhodamine B dimethylhydrazine (VII).
  • conjugate (XIII) useful as a probe for the location and isolation of receptor proteins that bind codeine and similar molecules.
  • the dodecamer peptide (BEAD)-Asp-His-Ile- Ala-Asn-Arg-Arg-Gly-Thr-Arg-Gly-Ser-NH 2 is obtained attached to the solid support as shown using standard FMOC peptide synthesis techniques, after deprotection of the terminal FMOC group.
  • This peptide is shaken with a solution of an equivalent molar amount of ClCH 2 COCl in a suitable solvent at 50 °C for 6 hours. The solvent is removed by decantation, leaving a terminal -NH-CO-CH 2 Cl group attached to the peptide.
  • This example teaches the synthesis of a competitive inhibitor for human elastase based on the structure of known N-trifluoroacetyl dipeptide analide inhibitors (see 162 J. Mol. Biol. 645 (1982) and
  • hydrazinium iodide enantiomer prepared as outlined below, 1.0 g (0.0106 mol) chloroacetic acid and 1.24 g (0.011 mol) chloracetyl chloride, contained in a micro reaction flask equipped with a drying tube, was heated in an oil bath at 105°C for 1 hour. The (homogeneous) reaction mixture was then cooled to room temperature and extracted with 4 ⁇ 20 ml of ethyl ether to remove chloracetyl chloride and chloracetic acid, with vigorous stirring each time. The residual semisolid was dissolved in the minimum amount of methanol and titrated with 10% KOH in methanol to the phenolphthalein end point. The
  • This example teaches the synthesis of a competitive inhibitor for the HIV protease with enhanced stability, based on the insertion of a chiral aminimide residue into the scissile bond position of the substrate
  • the side chain blocking groups are subsequently removed using standard peptide deprotection techniques to yield the product Ac-Ser-Leu-Asn-Phe-CON-N + (C 5 H 10 )-CH 2 -CO- NH-Val-Ile-OMe, useful as a enhanced stability
  • the functionalized silica was then collected by filtration, re-slurried in 100 ml methanol and refiltered a total of five times, then dried in a vacuum oven at 60°C/30" overnight to give 9.68 g of the product.
  • This functionalized silica was slurry packed from
  • This example describes preparation of an aminimide-functionalized ion-exchange silica matrix using epoxy silica as the support to be modified.
  • the reaction sequence is:
  • the diethylaminoethyl (DEAE) functionalized silica was collected by filtration, re-slurried in 100 ml methanol and re-filtered a total of five times. The packing was dried in a vacuum oven at 60°C/30" overnight. A 1.0 ml bed of this material was then packed in a 15 mM NaAc buffer at pH 7.7. The column was then equilibrated with 15 mM NaAc buffer at pH 5.6, and a solution of 1 mg/ml ovalbumin in this buffer run through the bed at a flow rate of 1.6 ml/min. A total of 59.2 ml of protein solution was run.
  • the column was then washed with 41.7 ml of 15 mM NaAc buffer at pH 5.58 and at a flow rate of 3.9 ml/min.
  • the bound protein was eluted using 23.4 ml of 0.5M NaCl at a flow rate of 3.9 ml/min.
  • the eluent (15.2 ml) was then collected and the transmission of an aliquot measured at 280 m ⁇ with a spectrophotometer.
  • ovalbumin concentration was determined from a calibration curve. The total amount of ovalbumin collected was 63.7 mg.
  • This example describes preparation of an aminimide-functionalized size-exclusion silica matrix using the epoxy silica support described above.
  • the mixture was allowed to stand at room temperature with periodic shaking for 45 min.
  • the functionalized silica was then collected by filtration, re-slurried in 100 ml methanol, re-filtered a total of five times and dried in a vacuum oven at 60°C/30" overnight.
  • the functionalized silica was slurry packed from methanol into a 10mm interior-diameter jacketed glass column with adjustable pistons to provide an 8 cm-long packed bed. This packing was used to separate mixtures of polyethylene glycol polymers of varying molecular weight with good resolution using a mobile phase.
  • Hydroxypropylcellulose is mono-functionalized by reaction, under strong alkaline conditions (preferably provided by a strong base, such as potassium t-butoxide) with C1CH 2 CON-N + (CH 3 ) 3 .
  • a strong base such as potassium t-butoxide
  • C1CH 2 CON-N + (CH 3 ) 3 The result is replacement of approximately one hydroxyl group in each saccharide unit with the aminimide as follows :
  • the resulting aminimide derivative is coated onto a surface (e.g., silica).
  • a surface e.g., silica.
  • the N(CH 3)3 group leaves, resulting in formation of an isocyanate moiety:
  • the isocyanate groups then react with unreacted hydroxyl groups on the saccharide units to produce a cross-linked coating.
  • the cellulose can be coated onto the surface and immobilized using standard techniques (e . g. , reaction with bisoxiranes), and then mono, di- or tri-substituted with desired aminimide derivatives as described above.
  • reaction sequence can also be employed with polymers or oligomers bearing NH or SH groups instead of hydroxyl groups and can also be utilized to fabricate structures such as crosslinked cellulose membranes.
  • This example illustrates an alternative immobilization technique, namely, polymerizing aminimide precursors containing vinyl groups and which have been coated onto a surface.
  • the chemistry resembles the approach described above, except polymerization forms a sturdy shell around an existing support rather than creating a solid block of material.
  • 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 coated silica was slurried in 100 ml of isooctane containing 86 mg of VAZO-64 dissolved in 1.5 ml toluene which had been de-aerated with nitrogen. The slurry was thoroughly de-aerated wtih nitrogen and then stirred at 70 °C for two hours.
  • coated silica was collected by filtration and washed three times in 100 ml methanol and air dried. The silica was heated at 120 °C for 2 hours to cure the coating. 13.1 g of coated silica were obtained. A 1 ml bed of this material was packed in an adjustable glass column and successfully used to separate BSA from
  • an epoxy-functionalized surface is reacted with disubstituted hydrazine, a bisepoxide and a triester to form a crosslinked network of aminimide chains attached covalently to the surface as follows:
  • the reaction can be carried out in water at room temperature without special conditions.
  • This example describes preparation of three-dimensional cross-linked porous copolymeric aminimide ion-exchange beads. It involves reaction of three monomers:
  • the beads obtained at the conclusion of the foregoing steps had a mean diameter of approximately 75 ⁇ and an ion-exchange capacity of 175 ⁇ eq/ml.
  • This example describes preparation of an aminimide electrophoresis gel.
  • the aminimide electrophoresis gel As a control, the
  • Tris 1.5M 6.06 g Tris base, 8 ml 10% SDS, volume adjusted to 90 ml with double-distilled water.
  • the pH was adjusted to 6.0 with concentrated HCl, and the final volume adjusted to 100 ml with DD water.
  • Ammonium persulfate 10% 0.1 g ammonium persulfate was dissolved in 0.9 ml DD water. The solution was used within 4 hours of preparation.
  • TMED used directly as obtained from Sigma Chemical Co., St. Louis, MO, under the tradename TMEDA.
  • This example describes preparation of latex particles containing an aminimide comonomer.
  • 591.1 ml of distilled water was charged to a three-necked round-bottomed flask.
  • a nitrogen dip tube was placed below the liquid level and the nitrogen flow rate set to 2 cm 3 /min.
  • the solution was mechanically agitated with a Teflon paddle at 250 RPM and heated to 80 °C over a half-hour period.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Analytical Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nanotechnology (AREA)
  • Biochemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Communicable Diseases (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Inorganic Chemistry (AREA)
  • Virology (AREA)
  • Manufacturing & Machinery (AREA)
  • Oncology (AREA)
  • Electrochemistry (AREA)
  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Saccharide Compounds (AREA)
  • Peptides Or Proteins (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)
  • Macromolecular Compounds Obtained By Forming Nitrogen-Containing Linkages In General (AREA)

Abstract

L'invention concerne la conception et la synthèse de nouveaux modules moléculaires à base d'aminimide ainsi que l'utilisation de ces modules dans l'élaboration de nouvelles molécules et de nouveaux matériaux fabriqués. Les nouvelles molécules et les nouveaux matériaux fabriqués constituent des agents de reconnaissance moléculaire utiles dans la conception et la synthèse de médicaments, et ils présentent des applications dans les sciences de séparation et des matéiaux.
EP93916884A 1992-06-30 1993-06-30 Molecules contenant des aminimides et materiaux utilises comme agents de reconnaissance moleculaire Withdrawn EP0723441A4 (fr)

Applications Claiming Priority (7)

Application Number Priority Date Filing Date Title
US90677092A 1992-06-30 1992-06-30
US90676992A 1992-06-30 1992-06-30
US906769 1992-06-30
US906770 1992-06-30
US4155993A 1993-04-02 1993-04-02
US41559 1993-04-02
PCT/US1993/006241 WO1994001102A1 (fr) 1992-06-30 1993-06-30 Molecules contenant des aminimides et materiaux utilises comme agents de reconnaissance moleculaire

Publications (2)

Publication Number Publication Date
EP0723441A1 true EP0723441A1 (fr) 1996-07-31
EP0723441A4 EP0723441A4 (fr) 1997-07-30

Family

ID=27365935

Family Applications (1)

Application Number Title Priority Date Filing Date
EP93916884A Withdrawn EP0723441A4 (fr) 1992-06-30 1993-06-30 Molecules contenant des aminimides et materiaux utilises comme agents de reconnaissance moleculaire

Country Status (7)

Country Link
EP (1) EP0723441A4 (fr)
JP (1) JPH08500339A (fr)
KR (1) KR950702113A (fr)
AU (1) AU685752B2 (fr)
BR (1) BR9306657A (fr)
CA (1) CA2139349A1 (fr)
WO (1) WO1994001102A1 (fr)

Families Citing this family (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5833665A (en) * 1990-06-14 1998-11-10 Integra Lifesciences I, Ltd. Polyurethane-biopolymer composite
US5795870A (en) * 1991-12-13 1998-08-18 Trustees Of Princeton University Compositions and methods for cell transformation
US5693769A (en) * 1991-12-13 1997-12-02 Transcell Technologies, Inc. Glycosylated steroid derivatives for transport across biological membranes and process for making and using same
AU689764B2 (en) * 1993-12-28 1998-04-09 Arqule, Inc. Modular design and synthesis of aminimide containing molecules
JPH09507487A (ja) * 1994-01-05 1997-07-29 アーキュール，インコーポレーテッド 選ばれた性質を有するアミンイミドおよびオキサゾロンをベースとした分子の系統的モジュール製造
US5734082A (en) * 1994-10-20 1998-03-31 Arqule Inc. Hydroxyethyl aminimides
US5807754A (en) * 1995-05-11 1998-09-15 Arqule, Inc. Combinatorial synthesis and high-throughput screening of a Rev-inhibiting arylidenediamide array
CA2231582A1 (fr) * 1995-10-17 1997-04-24 Genencor International, Inc. Composition enzymatique et son procede de preparation
US5874443A (en) * 1995-10-19 1999-02-23 Trega Biosciences, Inc. Isoquinoline derivatives and isoquinoline combinatorial libraries
US5840500A (en) * 1996-07-11 1998-11-24 Trega Biosciences, Inc. Quinoline derivatives and quinoline combinatorial libraries
US5916899A (en) * 1996-10-18 1999-06-29 Trega Biosciences, Inc. Isoquinoline derivatives and isoquinoline combinatorial libraries
US6617114B1 (en) 1996-10-31 2003-09-09 Karo Bio Ab Identification of drug complementary combinatorial libraries
WO1999011655A1 (fr) 1997-09-04 1999-03-11 Gryphon Sciences Banques modulaires de proteines et leurs procedes de preparation
US6784186B1 (en) * 1998-01-29 2004-08-31 Roy W. Jackson Therapeutic compounds
US6284735B1 (en) 1998-04-28 2001-09-04 Lion Bioscience Ag HP-3228 and related peptides to treat sexual dysfunction
US6534503B1 (en) 1998-04-28 2003-03-18 Lion Bioscience Ag Melanocortin receptor-3 ligands to treat sexual dysfunction
AU3768799A (en) * 1998-04-28 1999-11-16 Trega Biosciences, Inc. Isoquinoline compound melanocortin receptor ligands and methods of using same
FR2797632B1 (fr) * 1999-08-19 2003-01-24 Dutel Sa Fonctionnalisation d'un peptide par un groupement alpha- hydrazinoacetique
DE19946737A1 (de) * 1999-09-29 2001-04-19 Max Planck Gesellschaft Verwendung von Gangliosiden zur Modulation von Sphingolipid-Cholesterin-Mikrodomänen
KR101180201B1 (ko) * 2007-12-28 2012-09-05 미쓰이 가가쿠 가부시키가이샤 잠재성 경화제, 그것을 포함하는 에폭시 수지 조성물, 실링재 및 유기 ｅｌ 디스플레이
JP2013537966A (ja) 2010-08-31 2013-10-07 ジェネンテック， インコーポレイテッド バイオマーカー及び治療の方法
WO2012122512A1 (fr) 2011-03-10 2012-09-13 Hco Antibody, Inc. Production recombinante de mélanges d'anticorps monocaténaires
WO2015004028A1 (fr) 2013-07-08 2015-01-15 Bayer Cropscience Ag Dérivés de sulfures et sulfoxydes d'aryles hexacycliques à liaison c‑n utilisés comme pesticides
US10377928B2 (en) 2015-12-10 2019-08-13 Ppg Industries Ohio, Inc. Structural adhesive compositions
CN113171762B (zh) * 2021-04-27 2023-02-24 重庆科技学院 一种纤维基吸附材料及其制备方法和回收钯的应用
CN114247435B (zh) * 2021-12-25 2022-07-22 盐城工学院 一种高效吸附VOCs的有机变色材料的制备方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR1478953A (fr) * 1965-05-06 1967-04-28 Hoechst Ag Nouveaux benzène-sulfonyl-semicarbazides et leur préparation
FR1558886A (fr) * 1967-04-05 1969-02-28
DE1809950A1 (de) * 1968-11-20 1970-06-11 Basf Ag Substituierte Hydrazinderivate und diese enthaltende Mittel zur Regulierung des Pflanzenwachstums

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3671473A (en) * 1968-03-19 1972-06-20 Ashland Oil Inc Polymers formed from the reaction of a mixture of a polyepoxide and a polyester and an unsymmetrical disubstituted hydrazine
US3949140A (en) * 1974-05-06 1976-04-06 Owens-Corning Fiberglas Corporation Organo silicon derivatives coated on glass fibers
US3946131A (en) * 1974-05-06 1976-03-23 Owens-Corning Fiberglas Corporation Glass fibers coated with silicon containing aminimide compounds
US3898087A (en) * 1974-06-14 1975-08-05 Ball Corp Photopolymerizable compositions containing aminimides
US3985807A (en) * 1975-03-31 1976-10-12 Ashland Oil, Inc. Alkoxy derivatives of hydroxy aminimides
US4016340A (en) * 1975-08-07 1977-04-05 Polaroid Corporation Hydroxyl-containing polymers having aminimide groups attached thereto through an ether linkage
US4260705A (en) * 1976-06-30 1981-04-07 Board Of Regents, For And On Behalf Of The University Of Florida Addition copolymers of aminimides useful for affinity chromatography
US4330443A (en) * 1980-06-18 1982-05-18 The United States Of America As Represented By The Secretary Of Agriculture Dry chemical process for grafting acrylic and methyl acrylic ester and amide monomers onto starch-containing materials

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR1478953A (fr) * 1965-05-06 1967-04-28 Hoechst Ag Nouveaux benzène-sulfonyl-semicarbazides et leur préparation
FR1558886A (fr) * 1967-04-05 1969-02-28
DE1809950A1 (de) * 1968-11-20 1970-06-11 Basf Ag Substituierte Hydrazinderivate und diese enthaltende Mittel zur Regulierung des Pflanzenwachstums

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CANADIAN JOURNAL OF CHEMISTRY, vol. 45, 1967, OTTAWA CA, pages 2619-2623, XP002031748 W.J. MCKILLIP ET AL.: "Aminimides. II- A one-step synthesis of aminimides from carboxylic acid esters" *
JOURNAL OF THE CHEMICAL SOCIETY, PERKIN TRANSACTIONS 2, 1977, LETCHWORTH GB, pages 909-914, XP002031550 M. LILER ET AL. : "Mono- and di-protonation sites in N-ammonio-amidates: a spectroscopic study" *
See also references of WO9401102A1 *
SYNTHESIS, 1972, STUTTGART DE, pages 45-46, XP002031551 B. ZEEH ET AL. : "Eine einfache isoindol-synthese" *

Also Published As

Publication number Publication date
BR9306657A (pt) 1998-12-08
EP0723441A4 (fr) 1997-07-30
AU4659293A (en) 1994-01-31
WO1994001102A1 (fr) 1994-01-20
JPH08500339A (ja) 1996-01-16
AU685752B2 (en) 1998-01-29
CA2139349A1 (fr) 1994-01-20
KR950702113A (ko) 1995-06-19

Similar Documents

Publication Publication Date Title
WO1994001102A1 (fr) Molecules contenant des aminimides et materiaux utilises comme agents de reconnaissance moleculaire
US6271195B1 (en) Aminimide-containing molecules and materials as molecular recognition agents
US4965289A (en) Substrate and process for making a substrate
US4785021A (en) Adsorbent and method of preparation
JPH01151596A (ja) 結合オリゴヌクレオチド及びペプチドを有する膜
JP2002080530A (ja) 固相ペプチド合成のための樹脂
AU689764B2 (en) Modular design and synthesis of aminimide containing molecules
US7820785B2 (en) Solid phase synthesis supports and methods
WO1994000509A9 (fr) Matieres derivees d'oxazolone
AU678168B2 (en) Oxazolone derived materials
AU6084294A (en) Method of making polymers having specific properties
WO1995017903A1 (fr) Structure et synthese modulaire de molecules derivees de l'oxazolone
US20020146684A1 (en) One dimensional unichemo protection (UCP) in organic synthesis
CA2180527A1 (fr) Procede de preparation de polymeres ayant des proprietes specifiques
JPH06197B2 (ja) 吸着剤
CA2179984A1 (fr) Structure et synthese modulaire de molecules derivees de l'oxazolone

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 19950130

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LI LU MC NL PT SE

A4 Supplementary search report drawn up and despatched

Effective date: 19970611

AK Designated contracting states

Kind code of ref document: A4

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LI LU MC NL PT SE

17Q First examination report despatched

Effective date: 19980831

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: ARQULE, INC.

GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20040205