Methods and compositions for making antibodies and antibody derivatives with reduced core fucosylation

1. A mammalian cell culture medium for the production of antibodies or antibody derivatives having reduced core fucosylation, comprising an effective amount of a fucose analog, wherein the fucose analog is selected from the group consisting of one of the following formulae (III) or (IV):

or a biologically acceptable salt or solvate thereof, wherein each of formula (III) or (IV) can be the alpha or beta anomer or the corresponding aldose form;

each of R 1 -R 4 is independently selected from the group consisting of fluoro, chloro, —OH, —OC(O)H, —OC(O)C 1 -C 10 alkyl, —OC(O)C 2 -C 10 alkenyl, —OC(O)C 2 -C 10 alkynyl, —OC(O)aryl, —OC(O)heterocycle, —OC(O)C 1 -C 10 alkylene(aryl), —OC(O)C 2 -C 10 alkenylene(aryl), —OC(O)C 2 -C 10 alkynyl(aryl), —OC(O)C 1 -C 10 alkylene heterocycle, —OC(O)C 2 -C 10 alkenylene(heterocycle), —OC(O)C 2 -C 10 alkynyl heterocycle, —OCH 2 OC(O) alkyl, —OCH 2 OC(O)O alkyl, —OCH 2 OC(O) aryl, —OCH 2 OC(O)O aryl,

—OC(O)CH 2 O(CH 2 CH 2 O) n CH 3 , —OC(O)CH 2 CH 2 O(CH 2 CH 2 O) n CH 3 , —O-tri-C 1 -C 3 alkylsilyl and —OC 1 -C 10 alkyl, wherein each n is an integer independently selected from 0-5; and

each of R 2a and R 3a is independently selected from the group consisting of H, F and Cl;

R 5 is selected from the group consisting

of —CH 3 , —CHF 2 , —CH═C═CH 2 , —C≡CH, —C≡CCH 3 , —CH 2 C≡CH, —C(O)OCH 3 , —CH(OAc)CH 3 , —CN, —CH 2 CN, —CH 2 X (wherein X is Br, Cl or I), and methoxiran;

wherein when R 5 is other than —CH═C═CH 2 or —CHF 2 , at least one of R 1 , R 2 , R 3 , R 2a and R 3a is fluoro or chloro.

2. The culture medium of claim 1 wherein

each of R 1 —R 4 is independently selected from the group consisting of fluoro, chloro, —OH, —OC(O)H, —OC(O)C 1 -C 10 alkyl, —OC(O)C 2 -C 10 alkenyl, —OC(O)C 2 -C 10 alkynyl, —OC(O)aryl, —OC(O)heterocycle, —OC(O)C 1 -C 10 alkylene(aryl), —OC(O)C 2 -C 10 alkynyl(aryl), —OC(O)C 1 -C 10 alkylene heterocycle, —OC(O)C 2 -C 10 alkenylene(heterocycle), —OC(O)C 2 -C 10 alkynyl heterocycle, —OCH 2 OC(O) alkyl, —OCH 2 OC(O)O alkyl, —OCH 2 OC(O) aryl, —OCH 2 OC(O)O aryl, —OC(O)CH 2 O(CH 2 CH 2 O) n CH 3 , —OC(O)CH 2 CH 2 —O—(CH 2 CH 2 O) n CH 3 , —O-tri-C 1 -C 3 alkylsilyl and —OC 1 -C 10 alkyl, wherein each n is an integer independently selected from 0-5; and at least one of R 1 , R 2 , R 3 , R 2a and R 3a is fluoro or chloro.

3. The culture medium of claim 1 , which:

(i) is free of added animal protein;

(ii) is free of serum;

(iii) is free of added fucose; and

(iv) is a powder or a liquid.

4. The culture medium of claim 2 , which:

(i) is free of added animal protein;

(ii) is free of serum;

(iii) is free of added fucose; and

(iv) is a powder or a liquid.

5. The culture medium of claim 3 , wherein at least one of R 1 , R 2 , R 2a , R 3 and R 3a is fluoro.

6. The culture medium of claim 3 , wherein two of R 1 , R 2 , R 2a , R 3 and R 3a are fluoro or chloro.

7. The culture medium of claim 3 wherein the effective amount is an amount of the analog that is sufficient to decrease fucose incorporation into a complex N-glycoside-linked sugar chain of an antibody or antibody derivative by at least 90%.

8. The culture medium of claim 7 , wherein R 1 , R 3 and R 4 are each independently selected from —OH and —OAc; R 2 is F; R 2a and R 3a are each H; and R 5 is —CH 3 .

9. The culture medium of claim 3 , wherein the fucose analog is 2-deoxy-2-fluorofucose diacetate, 2-deoxy-2-chlorofucose triacetate, 2-deoxy-2-fluorofucose, 2-deoxy-2-fluorofucose peracetate, 1,2-difluoro-1,2-didexoy fucose peracetate, or 6,6-difluorofucose tetraacetate.

10. The culture medium of claim 3 , wherein the fucose analog is 2-deoxy-2-fluorofucose diacetate or 2-deoxy-2-chlorofucose triacetate.

11. The culture medium of claim 3 , wherein the fucose analog is 2-deoxy-2-fluorofucose.

12. The culture medium of claim 11 , wherein the effective amount is an amount of the analog that is sufficient to decrease fucose incorporation into a complex N-glycoside-linked sugar chain of an antibody or antibody derivative by at least 80%.

13. The culture medium of claim 12 , which is free of added fucose.

14. The culture medium of claim 1 wherein the antibody or antibody derivative is a humanized or human antibody or antibody derivative.

15. The culture medium of claim 1 wherein the mammalian cell culture medium is a Chinese hamster ovary cell culture medium.

16. The culture medium of claim 2 wherein the antibody or antibody derivative is a humanized or human antibody or antibody derivative.

17. The culture medium of claim 2 wherein the mammalian cell culture medium is a Chinese hamster ovary cell culture medium.

18. The culture medium of claim 7 wherein the antibody or antibody derivative is a humanized or human antibody or antibody derivative.

19. The culture medium of claim 7 wherein the mammalian cell culture medium is a Chinese hamster ovary cell culture medium.

20. The culture medium of claim 8 wherein the antibody or antibody derivative is a humanized or human antibody or antibody derivative.

21. The culture medium of claim 8 wherein the mammalian cell culture medium is a Chinese hamster ovary cell culture medium.

22. The culture medium of claim 11 wherein the antibody or antibody derivative is a humanized or human antibody or antibody derivative.

23. The culture medium of claim 11 wherein the mammalian cell culture medium is a Chinese hamster ovary cell culture medium.

24. The culture medium of claim 12 wherein the antibody or antibody derivative is a humanized or human antibody or antibody derivative.

25. The culture medium of claim 12 wherein the mammalian cell culture medium is a Chinese hamster ovary cell culture medium.

26. The culture medium of claim 13 wherein the antibody or antibody derivative is a humanized or human antibody or antibody derivative.

27. The culture medium of claim 13 wherein the mammalian cell culture medium is a Chinese hamster ovary cell culture medium.