Genetic package and uses thereof

• US 20090099031A1
• Filed: 03/06/2007
• Published: 04/16/2009
• Est. Priority Date: 09/27/2005
• Status: Abandoned Application

• Current Assignee: Amunix Operating Inc. (Sanofi )
• Original Assignee: Amunix Operating Inc. (Sanofi )
• Inventors: Schellenberger, Volker, Wang, Chia-wei, Stemmer, Willem P., Crameri, Andreas, Scholle, Michael D., Popkov, Mikhail, Gordon, Nathaniel C.
• Application Number:
  US11/715,300
• Publication Number:
  US 20090099031A1
• Time in Patent Office: Days
• US Class Current: 506/9
• 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: C07K 14/415 from plants

Abstract

The present invention provides unstructured recombinant polymers (URPs) and proteins containing one or more of the URPs. The present invention also provides microproteins, toxins and other related proteinaceous entities, as well as genetic packages displaying these entities. The present invention also provides recombinant polypeptides including vectors encoding the subject proteinaceous entities, as well as host cells comprising the vectors. The subject compositions have a variety of utilities including a range of pharmaceutical applications.

Claims

1. A genetic package displaying a microprotein, wherein said microprotein retains binding capability to its native target.

2. The genetic package of claim 1 wherein the microprotein exhibits binding capability towards at least one family of ion channel selected from the group consisting of a sodium, a potassium, a calcium, an acetylcholine, and a chlorine channel.

3. The genetic package of claim 1 wherein the microprotein is an ion-channel-binding microprotein, and is modified such that

(a) the microprotein binds to a different family of channel as compared to the corresponding unmodified microprotein;

(b) the microprotein binds to a different subfamily of the same channel family as compared to the corresponding unmodified microprotein;

(c) the microprotein binds to a different species of the same subfamily of channel as compared to the corresponding unmodified microprotein;

(d) the microprotein binds to a different site on the same channel as compared to the corresponding unmodified microprotein; and

/or

(e) the microprotein binds to the same site of the same channel but yield a different physiological effect as compared to the corresponding unmodified microprotein.

4. The genetic package of claim 1 wherein said microprotein is a toxin.

5. A library of genetic packages of any one of claims 1 through 4.

19. A method of obtaining a microprotein with desired property, comprising:

(a) providing a library of claim 5 or claim 6; and

(b) screening the selectable library to obtain at least one phage displaying a microprotein with the desired property.

20. The method of claim 19, wherein the screening the selectable library further comprises isolating the phage that displays the microprotein having the desired property.

21. The method of claim 19, wherein the desired property is binding specificity to a target of interest.

22. A recombinant polynucleotide comprising a coding sequence that encodes the protein claim 1 or 13.

23. A host cell comprising the recombinant polynucleotides of claim 22.

24. A vector comprising the recombinant polynucleotide of claim 22.

6. A genetic package displaying a proteinaceous toxin wherein said toxin retains in part or in whole its toxicity spectrum.

7. The genetic package of claim 6 wherein the toxin is a microprotein.

8. The genetic package of claim 6 wherein the toxin is derived from a single toxin protein, or derived from a family of toxins.

9. A library of genetic packages of claim 6, said library displaying a family of toxins, wherein the family retains in part or in whole its native toxicity spectrum.

10. The library of claim 9, wherein members of the family mediate toxicity via binding to an ion channel.

11. The library of claim 10, wherein the ion channel is selected from the group consisting of a sodium, potassium, calcium, acetylcholine, and chlorine channel.

12. The library of claim 9, wherein majority of the displayed members of the toxin family are substantially homologous.

13. A protein comprising a plurality of ion-channel binding domains, wherein individual domains are microprotein domains that have been modified such that

(a) the microprotein domains bind to a different family of channel as compared to the corresponding unmodified microprotein domains;

(b) the microprotein domains bind to a different subfamily of the same channel family as compared to the corresponding unmodified microprotein domains;

(c) the microprotein domains bind to a different species of the same subfamily as compared to the corresponding unmodified microprotein domains;

(d) the microprotein domains bind to a different site on the same channel as compared to the corresponding unmodified microprotein domains;

(e) the microprotein domains bind to the same site of the same channel but yield a different physiological effect as compared to the corresponding unmodified microprotein domains; and

/or

(f) the microprotein domains bind to the same site of the same channel and yield the same physiological effect as compared to the corresponding unmodified microprotein domains.

14. The protein of claim 13 wherein the microprotein domains comprise natural sequences.

15. The protein of claim 13 wherein the microprotein domains comprise non-natural sequences.

16. The protein of claim 13 wherein individual domains are linked together via a heterologous linker

17. The protein of claim 13 wherein individual microprotein domains bind to the same channel family, same channel subfamily, same species of the same subfamily, or same site on the same channel.

18. The protein of claim 13 wherein at least one individual microprotein domain binds to a different channel family, different channel subfamily, different species of the same subfamily, or different site on the same channel.