Hyperglycosylated polypeptide variants and methods of use

7,597,884

11/351163

February 08, 2006

The present invention provides synthetic Type I interferon receptor polypeptide agonists comprising consensus or hybrid Type I interferon receptor polypeptide agonists, containing one or more native or non-native glycosylation sites. The present invention provides synthetic Type I interferon receptor polypeptide agonists comprising consensus or hybrid Type I interferon receptor polypeptide agonists, containing one or more native or non-native glycosylation sites, as well as erythropoietin and darbepoetin alfa, each of which are linked to a penetrating peptide that facilitates translocation of a substance across a biological barrier as well as pharmaceutical compositions, including oral formulations, of the same. The present invention further provides oral formulations of hyperglycosylated or protease-resistant, hyperglycosylated polypeptide variants, which polypeptide variants lack at least one protease cleavage site found in a parent polypeptide, and thus exhibit increased protease resistance compared to the parent polypeptide, which polypeptide variants further include (1) a carbohydrate moiety covalently linked to at least one non-native glycosylation site not found in the parent protein therapeutic or (2) a carbohydrate moiety covalently linked to at least one native glycosylation site found but not glycosylated in the parent protein therapeutic. The present invention further provides compositions, including oral pharmaceutical compositions, comprising the synthetic Type I interferon receptor polypeptide agonist, the hyperglycosylated polypeptide variant, or the hyperglycosylated, protease-resistant polypeptide variant. The present invention further provides containers, devices, and kits comprising the synthetic Type I interferon receptor polypeptide agonist, the hyperglycosylated polypeptide variant, or the hyperglycosylated, protease-resistant polypeptide variant. The present invention further provides therapeutic methods involving administering an effective amount of an oral pharmaceutical composition comprising a synthetic Type I interferon receptor polypeptide agonist, a hyperglycosylated polypeptide variant, or a hyperglycosylated, protease-resistant polypeptide variant to an individual in need thereof.

Blatt, Lawrence M. (San Francisco, CA, US); Seiwert, Scott D. (Pacifica, CA, US); Hong, Jin (Brisbane, CA, US)

Alios BioPharma, Inc. (South San Francisco, CA, US)

1. A hyperglycosylated Type 1 interferon variant of a parent Type 1 interferon, wherein the parent Type 1 interferon is selected from the group consisting of interferon alfacon-1 and interferon α14, wherein the hyperglycosylated Type 1 interferon variant is the parent Type 1 interferon that has been modified to include at least three additional glycosylation sites, wherein at least one of the additional glycosylation sites is introduced by an amino acid substitution selected from the group consisting of D31N, D102N, D108N, and E138T.

2. The hyperglycosylated Type 1 interferon variant of claim 1 , wherein the parent Type 1 interferon is interferon alfacon-1.

3. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitution D102N.

4. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitution D108N.

5. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitution E138T.

6. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitutions D102N and D108N.

7. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitutions D102N and E138T.

8. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitutions D108N and E138T.

9. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitutions D102N, D108N, and E138T.

10. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is selected from the group consisting of an amino acid sequence set forth in SEQ ID NOS: 1769-1773.

11. The hyperglycosylated Type 1 interferon variant of claim 1 , wherein the parent Type 1 interferon is interferon α14.

12. The hyperglycosylated Type 1 interferon variant of claim 11 , wherein the hyperglycosylated Type 1 interferon variant is interferon α 14 comprising at least the amino acid substitution D108N.

13. The hyperglycosylated Type 1 interferon variant of claim 11 , wherein the hyperglycosylated Type 1 interferon variant is interferon α 14 comprising at least the amino acid substitution E138T.

14. The hyperglycosylated Type 1 interferon variant of claim 11 , wherein the hyperglycosylated Type 1 interferon variant is interferon α 14 comprising at least the amino acid substitution D108N and E138T.

15. A pharmaceutical composition comprising the hyperglycosylated Type 1 interferon variant of claim 1 ; and a pharmaceutically acceptable excipient.

16. The composition of claim 15 , wherein the pharmaceutically-acceptable excipient is suitable for oral delivery.

17. The composition of claim 15 , wherein the pharmaceutically-acceptable excipient is suitable for parenteral delivery.

18. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitutions D31N and D102N.

19. The hyperglycosylated Type 1 interferon variant of claim 3 , further comprising the amino acid substitution S104T.

20. The hyperglycosylated Type 1 interferon variant of claim 1 , further comprising the amino acid substitution S104T.

21. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitution D31N.

22. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitutions D31N and D108N.

23. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitutions D31N and E138T.

24. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitutions D31N, D102N, and D108N.

25. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitutions D31N, D102N, and E138T.

26. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitutions D31N, D108N, and E138T.

27. The hyperglycosylated Type 1 interferon variant of claim 2 , wherein the hyperglycosylated Type 1 interferon variant is interferon alfacon-1 comprising at least the amino acid substitutions D31N, D102N, D108N, and E138T.

424/854

3645090 February 1972 Mochizuki et al.
3720760 March 1973 Bennich et al.
3839346 October 1974 Gadekar
3940475 February 1976 Gross
3974281 August 1976 Gadekar
4052509 October 1977 Gadekar
4211771 July 1980 Witkowski et al.
RE30985 June 1982 Cartaya
4560655 December 1985 Baker
4657866 April 1987 Kumar
4683195 July 1987 Mullis et al.
4683202 July 1987 Mullis
4695623 September 1987 Stabinsky
4737456 April 1988 Weng et al.
4767704 August 1988 Cleveland et al.
4810643 March 1989 Souza
4810804 March 1989 Chandraratna
4877729 October 1989 Clark et al.
4897471 January 1990 Stabinsky
4927762 May 1990 Darfler
4959455 September 1990 Clark et al.
5041376 August 1991 Gething et al.
5122469 June 1992 Mather et al.
5194596 March 1993 Tischer et al.
5200534 April 1993 Rao
5202448 April 1993 Carver et al.
5229529 July 1993 Ueno et al.
5240848 August 1993 Keck et al.
5274137 December 1993 Nicolaou et al.
5279949 January 1994 Nair
5283253 February 1994 Holton et al.
5294637 March 1994 Chen et al.
5310562 May 1994 Margolin
5332671 July 1994 Ferrara et al.
5338840 August 1994 Bayne et al.
5395760 March 1995 Smith et al.
5415869 May 1995 Straubinger et al.
5426098 June 1995 Carlino
5462925 October 1995 Ogawa et al.
5466861 November 1995 Dawson et al.
5474978 December 1995 Bakaysa et al.
5484720 January 1996 Wurm et al.
5504188 April 1996 Baker et al.
5514646 May 1996 Chance et al.
5518729 May 1996 Margolin
5520911 May 1996 Anderson et al.
5521184 May 1996 Zimmermann
5532343 July 1996 Bayne et al.
5547929 August 1996 Anderson, Jr. et al.
5559101 September 1996 Weis et al.
5571714 November 1996 Dasch et al.
5593875 January 1997 Wurm et al.
5605690 February 1997 Jacobs et al.
5631015 May 1997 Bezwada et al.
5643575 July 1997 Martinez et al.
5650486 July 1997 De Felippis
5672662 September 1997 Harris et al.
5674843 October 1997 Carlino
5688666 November 1997 Bass et al.
5688679 November 1997 Powell
5690925 November 1997 Gray et al.
5693609 December 1997 Baker et al.
5700862 December 1997 Chance et al.
5712113 January 1998 Chung et al.
5716632 February 1998 Margolin
5726152 March 1998 Bayne et al.
5728074 March 1998 Castellano et al.
5728377 March 1998 Sarris et al.
5736371 April 1998 Samain et al.
5747642 May 1998 De Felippis
5759807 June 1998 Breece et al.
5770378 June 1998 Hwang et al.
5770382 June 1998 Hwang et al.
5770383 June 1998 Hwang et al.
5780676 July 1998 Boehm et al.
5783185 July 1998 Dasch et al.
5811395 September 1998 Schwabe et al.
5821263 October 1998 Scola et al.
5824685 October 1998 Campochiaro et al.
5824701 October 1998 Greenwald et al.
5853724 December 1998 Garrity et al.
5869680 February 1999 Mas et al.
5889144 March 1999 Alila et al.
5922675 July 1999 Baker et al.
5935567 August 1999 Leder et al.
5945402 August 1999 Cipolla et al.
5952297 September 1999 De Felippis et al.
5965697 October 1999 Czaplewski et al.
5985265 November 1999 Kinstler et al.
6022711 February 2000 Cunningham et al.
6034054 March 2000 DeFelippis et al.
6046305 April 2000 Choi
6057428 May 2000 Keyt et al.
6090822 July 2000 Margolin
6096010 August 2000 Walters et al.
6100071 August 2000 Davis-Smyth et al.
6127332 October 2000 Goelz et al.
6132970 October 2000 Stemmer
6136563 October 2000 Cunningham et al.
6143523 November 2000 Cunningham et al.
6143715 November 2000 Llinas-Brunet et al.
6146361 November 2000 DiBiasi et al.
6153600 November 2000 Leder et al.
6168784 January 2001 Offord et al.
6187330 February 2001 Wang et al.
6200953 March 2001 Schwabe et al.
6211144 April 2001 Havelund
6214542 April 2001 Striker et al.
6214854 April 2001 Wang et al.
6221053 April 2001 Walters et al.
6248095 June 2001 Giambattista et al.
6265380 July 2001 Tung et al.
6277830 August 2001 Ganguly et al.
6288089 September 2001 Zawada et al.
6296842 October 2001 Jaworowicz et al.
6299877 October 2001 Chen et al.
6300475 October 2001 Chen et al.
6303148 October 2001 Hennink et al.
6319921 November 2001 Cirillo et al.
6323180 November 2001 Llinas-Brunet et al.
6329379 December 2001 Llinas-Brunet et al.
6329417 December 2001 Llinas-Brunet et al.
6346269 February 2002 Hsiao et al.
6348444 February 2002 Chappel
6365157 April 2002 Rockwell et al.
6375929 April 2002 Thomas, Jr. et al.
6379701 April 2002 Tracy et al.
6383486 May 2002 Davis-Smyth et al.
6387879 May 2002 Blume et al.
6410531 June 2002 Llinas-Brunet et al.
6420380 July 2002 Llinas-Brunet et al.
6423695 July 2002 Tam et al.
6432962 August 2002 Horneman
6440985 August 2002 Bailey et al.
6448077 September 2002 Rockwell et al.
6458398 October 2002 Smith et al.
6475796 November 2002 Pollitt et al.
6479508 November 2002 Beaulieu et al.
6485942 November 2002 Zioncheck et al.
6489325 December 2002 Adams et al.
6491906 December 2002 Strieter et al.
6497871 December 2002 Gray et al.
6531122 March 2003 Pedersen et al.
6534523 March 2003 Llinas-Brunet et al.
6548520 April 2003 Adams et al.
6569420 May 2003 Chen et al.
6569871 May 2003 Adams et al.
6585398 July 2003 Haddad
6586398 July 2003 Kinstler et al.
6608027 August 2003 Tsantrizos et al.
6608067 August 2003 Tung et al.
6608182 August 2003 Rosen et al.
6617160 September 2003 Shitara et al.
6617309 September 2003 Tung et al.
6624290 September 2003 Zhang
6630155 October 2003 Chandrashekar et al.
6642204 November 2003 Llinas-Brunet et al.
6646110 November 2003 Nissen et al.
6673580 January 2004 Koren et al.
6685933 February 2004 Zoon et al.
6696056 February 2004 Cheung et al.
6703037 March 2004 Hubbell et al.
6703225 March 2004 Kojima et al.
6709649 March 2004 Lusso et al.
6730303 May 2004 Feng et al.
2001/0036955 November 2001 Gerritsen et al.
2002/0058635 May 2002 Averett
2002/0156023 October 2002 Walling et al.
2002/0183364 December 2002 Tang
2003/0064069 April 2003 Thompson et al.
2003/0073832 April 2003 Havez
2003/0091566 May 2003 Thompson et al.
2003/0133907 July 2003 Van Den Hazel et al.
2003/0149041 August 2003 Erickson et al.
2003/0153046 August 2003 Jensen et al.
2003/0186895 October 2003 Llinas-Brunet et al.
2003/0187018 October 2003 Llinas-Brunet et al.
2004/0023869 February 2004 Sims et al.
2004/0132977 July 2004 Gantier et al.
2006/0182716 August 2006 Hong et al.
640619 March 1995
WO 00/26354 May 2000
WO 01/36001 May 2001
WO 02/081507 October 2002
WO 03/066859 August 2003
WO 03/075944 September 2003
WO 2004/019856 March 2004
WO 2004/022593 March 2004
WO 2004/022747 March 2004
WO 2004/031352 April 2004

Infergen from www.rxlist.com/cgi/generic/infergen.htm (enclosed). cited by examiner
Infergen from www.drugdigest.org/DD/DVH/Uses/0,3915,891|Infergen,00.html (enclosed). cited by examiner
NCBI Accession No. CAA80408. cited by examiner
Loutfy MR, Blatt LM, Siminovitch KA, Ward S, Wolff B, Lho H, Pham DH, Deif H, LaMere EA, Chang M, Kain KC, Farcas GA, Ferguson P, Latchford M, Levy G, Dennis JW, Lai EKY, Fish EN, Interferon ALfacon-1 Plus Corticosteroids in Severe Acute Respriatory Syndrome, JAMA, 2003, 290(24): 3222-3228. cited by examiner
Keating MR, Antiviral Agents for Non-Human Immunodeficiency VIrus Infections, Mayo CLinic Proceeding, 1999, 74: 1226-1283 (as enclosed, pp. 1-26). cited by examiner
Nyman TA, Tolo H, Parkkinen J, Kalkkinen N, Identification of nine interferon-a subtypes produced by Sendai virus-induced human peripheral blood leucocytes, Biochem J., 1998, 329: 295-302. cited by examiner
GenBank Accession # NP—002163. cited by examiner
GenBank Accession No. NP—990758. Accessed Jul. 30, 2008. cited by examiner
GenBank Accession No. AAB27160. Accessed Jul. 30, 2008. cited by examiner
Egrie and Brown, Br J Cancer. Apr. 2001;84 Suppl 1:3-10. cited by other
Nyman et al. (1998) Eur. J. Biochem. 253:485-493. cited by other
Runkel et al. (1998) Pharmaceutical Research 15:641. cited by other
Adolf et al. (1990) J. Biol. Chem. 265:9290-9295. cited by other
International Search Report dated Nov. 23, 2007 for PCT/US2007/003333. cited by other
Written Opinion of the International Searching Authority dated Nov. 23, 2007 for PCT/US2007/003333. cited by other
Office Action dated Jun. 20, 2008 for U.S. Appl. No. 11/330,917, Filed Jan. 11, 2006. cited by other
International Search Report and Written Opinion dated Aug. 22, 2006 for International Application No. PCT/US2005/028165, Filed Aug. 8, 2005. cited by other
International Preliminary Report on Patentability dated Feb. 13, 2007 for International Application No. PCT/US2005/028165, Filed Aug. 8, 2005. cited by other
International Search Report and Written Opinion (Corrected Version) dated Oct. 26, 2007 for International Application No. PCT/US2007/003333, Filed Feb. 7, 2007. cited by other
International Preliminary Report on Patentability dated Aug. 8, 2008 for PCT/US2007/003333, Filed Feb. 7, 2007. cited by other

edspgr.07597884