Modified peptide and peptide libraries with protease resistance, derivatives thereof and methods of producing and screening such

• US 5,965,695 A
• Filed: 05/10/1995
• Issued: 10/12/1999
• Est. Priority Date: 05/15/1990
• Status: Expired due to Term

• Current Assignee: Chiron Corporation (Bayer AG)
• Original Assignee: Chiron Corporation (Bayer AG)
• Inventors: Simon, Reyna J., Bartlett, Paul A., Santi, Daniel V.
• Primary Examiner(s): Tsang, Cecilia J.
• Assistant Examiner(s): Lukton, David
• Application Number:
  US08/438,896
• Time in Patent Office: 1,616 Days
• Field of Search: 514/2, 514/12, 514/13-17, 530/323, 530/324, 530/325, 530/332, 530/345
• US Class Current: 530/324
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 47/62 the modifying agent being a protein, peptide or polyamino acid
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00283 Reactor vessels with top opening
• CPC Class Codes: B01J 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00308 interchangeably mounted in racks or blocks
• CPC Class Codes: B01J 2219/00416 Vacuum
• CPC Class Codes: B01J 2219/00418 using pressure
• CPC Class Codes: B01J 2219/00423 using filtration, e.g. through porous frits
• CPC Class Codes: B01J 2219/00466 in a slurry
• CPC Class Codes: B01J 2219/00493 by sparging or bubbling with gases
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00592 Split-and-pool, mix-and-divide processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00691 using robots
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07K 1/045 : using devices to improve synthesis, e.g. reactors, special vessels
• CPC Class Codes: C07K 1/047 : Simultaneous synthesis of different peptide species; Peptide libraries
• CPC Class Codes: C07K 14/001 : by chemical synthesis
• CPC Class Codes: C07K 5/0207 : containing the structure -NH-(X)4-C(=0), e.g. 'isosters', replacing two amino acids
• CPC Class Codes: C07K 7/02 : Linear peptides containing at least one abnormal peptide link
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries

Abstract

Peptoids are provided which are polymers comprised of monomer units wherein the monomer units include least some substitute amino acids and may include conventional amino acids. The peptoids can be synthesized in large numbers so as to provide libraries of peptoids which can be screened in order to isolate peptoids of desired biological activity. Although the peptoids may include amino acids, they preferably include only substituted amino acids and are designed in a manner so as to have a particular biological activity. Certain peptoids are designed to mimic as closely as possible the activity of known proteins. Other peptoids are designed so as to have greater or lesser activity than known proteins and may be designed so as to block known receptor sites and/or elicit a desired immunogenic response and thereby act as vaccines. In that the peptoids are comprised of substitute amino acids they can be designed to have structures which natural proteins cannot conform to. A range of different amino acid substitutes are disclosed, as are their methods of synthesis and methods of using such amino acid substitutes in the synthesis of peptoids and peptoid libraries. Methods of screening the libraries in order to obtain desired peptoids of a particular biological activity are also disclosed. The peptoids are preferably linked to a pharmaceutically active drug forming a conjugate with increased binding affinity to a particular biological receptor site.

Claims

1. A peptoid conjugate consisting of a pharmaceutically active drug covalently bound to a heteropolymeric peptoid compound consisting of 2 to 50 monomeric units linked by peptide bonds, wherein said monomeric units are selected from the group consisting of amino acid residues and at least 2 different N-substituted glycine residues having an α

-amino R group and differing by R group, wherein the α

-amino R group of each of said N-substituted glycine residues is alkyl of 2-6 carbon atoms, haloalkyl of 1-6 carbon atoms where halo is F, Cl, Br, or I, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkoxyalkyl of 2-8 carbon atoms, aryl of 6-10 carbon atoms, arylalkyl of 7-12 carbon atoms, arylalkyl of 7-12 carbon atoms substituted with 1-3 radicals independently selected from the group consisting of halo and nitro and hydroxy, aminoalkyl of 1-6 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, carboxy, carboxyalkyl of 2-6 carbon atoms, carboalkoxy-alkyl of 3-10 carbon atoms, carbamyl, carbamylalkyl of 2-6 carbon atoms, guanidino, guanidinoalkyl of 1-6 carbon atoms, mercapto, mercaptoalkyl of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, alkylthioalkyl of 2-10 carbon atoms, imidazolyl, imidazolylalkyl of 4-10 carbon atoms, pyridyl, pyridylalkyl of 6-10 carbon atoms, piperidyl, piperidylalkyl of 5-10 carbon atoms, indolyl, or indolylalkyl of 9-15 carbon atoms, wherein less than 50% of said monomeric units are amino acids and at least two of said monomeric units are different N-substituted glycine residues that differ by α

-amino R group.

2. A peptoid conjugate compound as claimed in claim 1, wherein the pharmaceutically active drug is a steroid.

3. A peptoid conjugate compound as claimed in claim 1, wherein the conjugate has a greater binding affinity with a bioactive receptor site than either the peptoid or the drug.

4. A peptoid conjugate compound as claimed in claim 1, wherein said pharmaceutically active drug is an antibacterial agent.

5. A peptoid conjugate compound as claimed in claim 4, wherein said antibacterial agent is a dihydrofolate reductase inhibitor.

6. A peptoid conjugate compound as claimed in claim 5, wherein said dihydrofolate reductase inhibitor is trimethoprim.

7. A peptoid conjugate compound as claimed in claim 5, wherein said dihydrofolate reductase inhibitor is 2,4-diamino-5-[3,5-di-methoxy-4-(3-hydroxycarboxy-1-oxopropylamino)benzyl]pyrimidine.

8. A peptoid conjugate compound as claimed in claim 1, wherein said pharmaceutically active drug is AZT.

9. The peptoid conjugate of claim 1, wherein all of said monomeric units are N-substituted glycine residues.

10. A peptoid conjugate consisting of a pharmaceutically active drug covalently bound to a heteropolymeric peptoid compound consisting of 2 to 50 monomeric units linked by peptide bonds, wherein said monomeric units are selected from N-substituted glycine residues having an α

-amino R group and differing by R group, wherein the α

-amino R group of each of said N-substituted glycine residues is alkyl of 2-6 carbon atoms, haloalkyl of 1-6 carbon atoms where halo is F, Cl, Br, or I, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkoxyalkyl of 2-8 carbon atoms, aryl of 6-10 carbon atoms, arylalkyl of 7-12 carbon atoms, arylalkyl of 7-12 carbon atoms substituted with 1-3 radicals independently selected from the group consisting of halo and nitro and hydroxy, aminoalkyl of 1-6 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, carboxy, carboxyalkyl of 2-6 carbon atoms, carboalkoxy-alkyl of 3-10 carbon atoms, carbamyl, carbamylalkyl of 2-6 carbon atoms, guanidino, guanidinoalkyl of 1-6 carbon atoms, mercapto, mercaptoalkyl of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, alkylthioalkyl of 2-10 carbon atoms, imidazolyl, imidazolylalkyl of 4-10 carbon atoms, pyridyl, pyridylalkyl of 6-10 carbon atoms, piperidyl, piperidylalkyl of 5-10 carbon atoms, indolyl, or indolylalkyl of 9-15 carbon atoms;

wherein at least two of said monomeric units are different N-substituted glycine residues that differ by α

-amino R group.

11. A peptoid conjugate compound as claimed in claim 10, wherein the pharmaceutically active drug is a steroid.

12. A peptoid conjugate compound as claimed in claim 10, wherein the conjugate has a greater binding affinity with a bioactive receptor site than either the peptoid or the drug.

13. A peptoid conjugate compound as claimed in claim 10, wherein said pharmaceutically active drug is an antibacterial agent.

14. A peptoid conjugate compound as claimed in claim 13, wherein said antibacterial agent is a dihydrofolate reductase inhibitor.

15. A peptoid conjugate compound as claimed in claim 14, wherein said dihydrofolate reductase inhibitor is trimethoprim.

16. A peptoid conjugate compound as claimed in claim 14, wherein said dihydrofolate reductase inhibitor is 2,4-diamino-5-[3,5-di-methoxy-4-(3-hydroxycarboxy-1-oxopropylamino)benzyl]pyrimidine.

17. A peptoid conjugate compound as claimed in claim 10, wherein said pharmaceutically active drug is AZT.