Claims for Patent: 8,937,161

Summary for Patent: 8,937,161

Title:	Cysteine engineered anti-TENB2 antibodies and antibody drug conjugates
Abstract:	Cysteine engineered anti-TENB2 antibodies are engineered by replacing one or more amino acids of a parent anti-TENB2 antibody with non cross-linked, reactive cysteine amino acids. Methods of design, preparation, screening, and selection of the cysteine engineered anti-TENB2 antibodies are provided. Cysteine engineered anti-TENB2 antibodies (Ab) are conjugated with one or more drug moieties (D) through a linker (L) to form cysteine engineered anti-TENB2 antibody-drug conjugates having Formula I: Ab-(L-D).sub.p I where p is 1 to 4. Diagnostic and therapeutic uses for cysteine engineered antibody drug compounds and compositions are disclosed.
Inventor(s):	Mao; Weiguang (San Mateo, CA), Junutula; Jagath Reddy (Fremont, CA), Polakis; Paul (Mill Valley, CA)
Assignee:	Genentech, Inc. (South San Francisco, CA)
Application Number:	12/288,181
Patent Claims:	1. A cysteine engineered anti-TENB2 antibody comprising a heavy and a light chain, one or more free cysteine amino acid residues located in the heavy chain, and the sequence: TABLE-US-00003 Sequence near Sequential Kabat Seq. Cys mutation Numbering Numbering I.D. VTVSSCSTKGP A121C A114C 12 in combination with one or more additional cysteine mutations in the heavy chain, selected from the group consisting of: TABLE-US-00004 Sequence near Sequential Kabat Seq. Cys mutation Numbering Numbering I.D. DVQLCESGPG Q5C Q5C 8 LSLTCCVSGYS A23C A23C 9 VSSASCKGPSV T123C T116C 13 WYVDGCEVHNA V285C V278C 14 KGFYPCDIAVE S378C S371C 15 PPVLDCDGSFF S403C S396C 16. 2. A cysteine engineered anti-TENB2 antibody comprising a heavy and a light chain, wherein said antibody comprises one or more free cysteine amino acid residues located in the heavy chain, and cysteine mutations: TABLE-US-00005 Sequence near Sequential Kabat Seq. Cys mutation Numbering Numbering I.D. VTVSSCSTKGP A121C A114C 12 and TABLE-US-00006 Sequence near Sequential Kabat Seq. Cys mutation Numbering Numbering I.D. DVQLCESGPG Q5C Q5C 8. 3. The cysteine engineered anti-TENB2 antibody of claim 1 or claim 2 prepared by a process comprising replacing one or more amino acid residues of a parent anti-TENB2 antibody by cysteine. 4. The cysteine engineered anti-TENB2 antibody of claim 1 or claim 2 further comprising one or more free cysteine amino acid residues located in the light chain. 5. The cysteine engineered anti-TENB2 antibody of claim 4 comprising in the light chain one or more sequences selected from: TABLE-US-00007 Sequence near Sequential Kabat Seq. Cys mutation Numbering Numbering I.D. SLSASCGDRVT V15C Vl5C 17 EIKRTCAAPSV V110C V110C 18 TVAAPCVFIFP S114C S114C 19 FIFPPCDEQLK S121C Sl2lC 20 DEQLKCGTASV S127C S127C 21 VTEQDCKDSTY S168C S168C 22 GLSSPCTKSFN V205C V205C 23. 6. The cysteine engineered anti-TENB2 antibody of claim 1 or claim 2 which is engineered from a parent anti-TENB2 antibody selected from a monoclonal antibody, a bispecific antibody, a chimeric antibody, a human antibody, and a humanized antibody. 7. The cysteine engineered anti-TENB2 antibody of claim 1 or claim 2 which is an antibody fragment. 8. The cysteine engineered anti-TENB2 antibody of claim 7 wherein the antibody fragment is a Fab fragment. 9. The cysteine engineered anti-TENB2 antibody of claim 1 or claim 2 which is selected from a chimeric antibody, a human antibody, or a humanized antibody. 10. The cysteine engineered anti-TENB2 antibody of claim 1 or claim 2 which is produced in bacteria. 11. The cysteine engineered anti-TENB2 antibody of claim 1 or claim 2 which is produced in CHO cells. 12. A pharmaceutical formulation comprising the cysteine engineered anti-TENB2 antibody of claim 1 or claim 2, and a pharmaceutically acceptable diluent, carrier or excipient. 13. The cysteine engineered anti-TENB2 antibody of claim 1 or claim 2 wherein the antibody is covalently attached to an auristatin drug moiety whereby an antibody drug conjugate is formed. 14. The antibody-drug conjugate of claim 13 comprising a cysteine engineered anti-TENB2 antibody (Ab), and an auristatin drug moiety (D) wherein the cysteine engineered anti-TENB2 antibody is attached through one or more free cysteine amino acids by a linker moiety (L) to D; the compound having Formula I: Ab-(L-D).sub.p I where p is 1, 2, 3, or 4. 15. The antibody-drug conjugate compound of claim 14 wherein p is 2. 16. The antibody-drug conjugate compound of claim 14 wherein L has the formula: -A.sub.a-W.sub.w--Y.sub.y-- where: A is a Stretcher unit covalently attached to a cysteine thiol of the cysteine engineered antibody (Ab); a is 0 or 1; each W is independently an Amino Acid unit; w is an integer ranging from 0 to 12; Y is a Spacer unit covalently attached to the drug moiety; and y is 0, 1 or 2. 17. The antibody-drug conjugate compound of claim 16 having the formula: ##STR00027## where PAB is para-aminobenzylcarbamoyl, and R.sup.17 is a divalent radical selected from (CH.sub.2).sub.r, C.sub.3-C.sub.8 carbocyclyl, O--(CH.sub.2).sub.r, arylene, (CH.sub.2).sub.r-arylene, -arylene-(CH.sub.2).sub.r--, (CH.sub.2).sub.r--(C.sub.3-C.sub.8 carbocyclyl), (C.sub.3-C.sub.8 carbocyclyl)-(CH.sub.2).sub.r, C.sub.3-C.sub.8 heterocyclyl, (CH.sub.2).sub.r--(C.sub.3-C.sub.8 heterocyclyl), --(C.sub.3-C.sub.8 heterocyclyl)-(CH.sub.2).sub.r--, --(CH.sub.2).sub.rC(O)NR.sup.b(CH.sub.2).sub.r--, --(CH.sub.2CH.sub.2O).sub.r--, --(CH.sub.2CH.sub.2O).sub.r--CH.sub.2--, --(CH.sub.2).sub.rC(O)NR.sup.b(CH.sub.2CH.sub.2O).sub.r--, --(CH.sub.2).sub.rC(O)NR.sup.b(CH.sub.2CH.sub.2O).sub.r--CH.sub.2--, --(CH.sub.2CH.sub.2O).sub.rC(O)NR.sup.b(CH.sub.2CH.sub.2O).sub.r--, --(CH.sub.2CH.sub.2O).sub.rC(O)NR.sup.b(CH.sub.2CH.sub.2O).sub.r--CH.sub.- 2--, and --(CH.sub.2CH.sub.2O).sub.rC(O)NR.sup.b(CH.sub.2).sub.r--; where R.sup.b is H, C.sub.1-C.sub.6 alkyl, phenyl, or benzyl; and r is independently an integer ranging from 1 to 10. 18. The antibody-drug conjugate compound of claim 16 wherein W.sub.w is valine-citrulline. 19. The antibody-drug conjugate compound of claim 17 wherein R.sup.17 is (CH.sub.2).sub.5 or (CH.sub.2).sub.2. 20. The antibody-drug conjugate compound of claim 16 having the formula: ##STR00028## 21. The antibody-drug conjugate compound of claim 20 wherein R.sup.17 is (CH.sub.2).sub.5 or (CH.sub.2).sub.2. 22. The antibody-drug conjugate compound of claim 16 having the formula: ##STR00029## 23. The antibody-drug conjugate compound of claim 14 wherein L is SMCC or BMPEO. 24. The antibody-drug conjugate compound of claim 14 wherein D is MMAE, having the structure: ##STR00030## where the wavy line indicates the attachment site to the linker L. 25. The antibody-drug conjugate compound of claim 14 wherein D is MMAF, having the structure: ##STR00031## where the wavy line indicates the attachment site to the linker L. 26. The antibody-drug conjugate compound of claim 13 wherein the parent anti-TENB2 antibody is selected from a monoclonal antibody, a bispecific antibody, a chimeric antibody, a human antibody, and a humanized antibody. 27. The antibody-drug conjugate compound of claim 13 which is engineered from a parent anti-TENB2 antibody that is an antibody fragment. 28. The antibody-drug conjugate compound of claim 27 wherein the antibody fragment is a Fab fragment. 29. The antibody drug conjugate of claim 13 wherein the auristatin is MMAE or MMAF. 30. The antibody drug conjugate of claim 13 wherein L is MC-val-cit-PAB or MC. 31. The antibody drug conjugate of claim 13 wherein L is SMCC, SPP, or BMPEO. 32. An antibody-drug conjugate compound selected from the structures: ##STR00032## wherein Val is valine; Cit is citrulline; p is 1, 2, 3, or 4; and Ab is a cysteine engineered anti-TENB2 antibody of claim 1. 33. The antibody drug conjugate of claim 32 wherein Ab comprises SEQ ID NO:2. 34. A pharmaceutical formulation comprising the antibody drug conjugate of claim 13, and a pharmaceutically acceptable diluent, carrier or excipient. 35. The pharmaceutical formulation of claim 34 further comprising a therapeutically effective amount of a chemotherapeutic agent selected from letrozole, oxaliplatin, doxetaxel, 5-FU, lapatinib, capecitabine, leucovorin, erlotinib, pertuzumab, bevacizumab, and gemcitabine. 36. An article of manufacture comprising the pharmaceutical formulation of claim 34; a container; and a package insert or label indicating that the compound can be used to treat cancer characterized by the overexpression of a TENB2 polypeptide. 37. The article of manufacture of claim 36 wherein the cancer is ovarian cancer, prostate cancer, cancer of the urinary tract, pancreatic cancer, lung cancer, breast cancer, or colon cancer.

Details for Patent 8,937,161

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genentech, Inc.	AVASTIN	bevacizumab	Injection	125085	02/26/2004		2027-10-19
Genentech, Inc.	PERJETA	pertuzumab	Injection	125409	06/08/2012		2027-10-19