ANTI-CD79B ANTIBODIES AND IMMUNOCONJUGATES AND METHODS OF USE

1 - 258 . (canceled)

259 . An antibody-drug conjugate comprising an anti-CD79b cysteine engineered antibody comprising a heavy and a light chain, a single free cysteine amino acid at position 118 of the heavy chain according to EU numbering convention or 205 of the light chain according to Kabat numbering convention, and comprises CDRs selected from

(a)

(i) an HVR-L1 sequence of

(SEQ ID NO: 194)

KASQSVDYEGDSFLN;

(ii) an HVR-L2 sequence of

(SEQ ID NO: 195)

AASNLES;

(iii) an HVR-L3 sequence of

(SEQ ID NO: 196)

QQSNEDPLT;

(iv) an HVR-H1 sequence of

(SEQ ID NO: 202)

GYTFSSYWIE;

(v) an HVR-H2 sequence of

(SEQ ID NO: 203)

GEILPGGGDTNYNEIFKG;

and

(vi) an HVR-H3 sequence of

(SEQ ID NO: 204)

TRRVPIRLDY;

(b)

(i) an HVR-L1 sequence of

(SEQ ID NO: 156)

KASQSVDYDGDSFLN;

(ii) an HVR-L2 sequence of

(SEQ ID NO: 157)

AASNLES;

(iii) an HVR-L3 sequence of

(SEQ ID NO: 158)

QQSNEDPLT;

(iv) an HVR-H1 sequence of

(SEQ ID NO: 164)

GYTFSSYWIE;

(v) an HVR-H2 sequence of

(SEQ ID NO: 165)

GEILPGGGDTNYNEIFKG;

and

(vi) an HVR-H3 sequence of

(SEQ ID NO: 166)

TRRVPVYFDY;

(c)

(i) an HVR-L1 sequence of

(SEQ ID NO: 175)

KASQSVDYDGDSFLN;

(ii) an HVR-L2 sequence of

(SEQ ID NO: 176)

AASNLES;

(iii) an HVR-L3 sequence of

(SEQ ID NO: 177)

QQSNEDPLT;

(iv) an HVR-H1 sequence of

(SEQ ID NO: 183)

GYTFSSYWIE;

(v) an HVR-H2 sequence of

(SEQ ID NO: 184)

GEILPGGGDTNYNEIFKG;

and

(vi) an HVR-H3 sequence of

(SEQ ID NO: 185)

TRRVPIRLDY;

and

(d)

(i) an HVR-L1 sequence of

(SEQ ID NO: 213)

KASQSVDYSGDSFLN;

(ii) an HVR-L2 sequence of

(SEQ ID NO: 176)

AASNLES;

(iii) an HVR-L3 sequence of

(SEQ ID NO: 177)

QQSNEDPLT;

(iv) an HVR-H1 sequence of

(SEQ ID NO: 221)

GYTFSSYWIE;

(v) an HVR-H2 sequence of

(SEQ ID NO: 222)

GEILPGGGDTNYNEIFKG;

and

(vi) an HVR-H3 sequence of

(SEQ ID NO: 223)

TRRVPIRLDY

wherein said antibody is covalently attached to a cytotoxic agent.

260 . The antibody-drug conjugate according to claim 259 , wherein the cytotoxic agent is selected from a chemotherapeutic agent, a growth inhibitory agent, a toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or fragments thereof), a drug moiety, an antibiotic, a nucleolytic enzyme and a radioactive isotope (i.e., a radioconjugate).

261 . The antibody-drug conjugate according to claim 260 , wherein the cytotoxic agent is a chemotherapeutic agent.

262 . The antibody-drug conjugate according to claim 260 , wherein the cytotoxic agent is a growth inhibitory agent

263 . The antibody-drug conjugate according to claim 260 , wherein the cytotoxic agent is a toxin.

264 . The antibody-drug conjugate according to claim 260 , wherein the cytotoxic agent is a radioactive isotope.

265 . The antibody-drug conjugate of claim 260 wherein the drug moiety is an auristatin or a maytansinoid.

266 . The antibody-drug conjugate of claim 265 comprising an antibody (Ab), and an auristatin or maytansinoid drug moiety (D) wherein the cysteine engineered antibody is attached through one or more free cysteine amino acids by a linker moiety (L) to D; the compound having Formula I:

A b -(L-D) p   I

where p is 1, 2, 3, or 4.

267 . The antibody-drug conjugate of claim 266 wherein p is 2.

268 . The antibody-drug conjugate of claim 266 wherein L has the formula:

-A a -W w —Y 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.

269 . The antibody-drug conjugate of claim 268 having the formula:

where PAB is para-aminobenzylcarbamoyl, and R 17 is a divalent radical selected from (CH 2 ) r , C 3 -C 8 carbocyclyl, O—(CH 2 ) r , arylene, (CH 2 ) r -arylene, -arylene-(CH 2 ) r —, (CH 2 ) r —(C 3 -C 8 carbocyclyl), (C 3 -C 8 carbocyclyl)-(CH 2 ) r , C 3 -C 8 heterocyclyl, (CH 2 ) r —(C 3 -C 8 heterocyclyl), —(C 3 -C 8 heterocyclyl)-(CH 2 ) r —, —(CH 2 ) r C(O)NR b (CH 2 ) r —, —(CH 2 CH 2 O) r —, —(CH 2 CH 2 O) r —CH 2 —, —(CH 2 ) r C(O)NR b (CH 2 CH 2 O) r —, —(CH 2 ) r C(O)NR b (CH 2 CH 2 O) r —CH 2 —, —(CH 2 CH 2 O) r C(O)NR b (CH 2 CH 2 O) r —, —(CH 2 CH 2 O) r C(O)NR b (CH 2 CH 2 O) r —CH 2 —, and —(CH 2 CH 2 O) r C(O)NR b (CH 2 ) r —; where R b is H, C 1 -C 6 alkyl, phenyl, or benzyl; and r is independently an integer ranging from 1 to 10.

270 . The antibody-drug conjugate of claim 269 wherein W w is valine-citrulline.

271 . The antibody-drug conjugate of claim 269 wherein R 17 is (CH 2 ) 5 or (CH 2 ) 2 .

272 . The antibody-drug conjugate of claim 268 having the formula:

273 . The antibody-drug conjugate of claim 272 wherein R 17 is (CH 2 ) 5 or (CH 2 ) 2 .

274 . The antibody-drug conjugate of claim 272 having the formula:

275 . The antibody-drug conjugate of claim 268 wherein L comprises 4-((2,5-dioxopyrrolidin-1-yl)methyl)cyclohexanecarboxylic acid (SMCC), 4-mercaptopentanoic acid (SPP), 4-mercaptobutanoic acid (SPDB) or Bis-Maleimido polyethylene glycol (BMPEO).

276 . The antibody-drug conjugate of claim 268 wherein D is monomethyl auristatin E (MMAE), having the structure:

wherein the wavy line indicates the attachment site to the linker L.

277 . The antibody-drug conjugate of claim 268 wherein D is monomethyl auristatin F (MMAF), having the structure:

wherein the wavy line indicates the attachment site to the linker L.

278 . The antibody-drug conjugate of claim 268 wherein D is DM1 or DM4, having the structures:

wherein the wavy line indicates the attachment site to the linker L.

279 . The antibody-drug conjugate of claim 265 wherein the antibody is selected from a monoclonal antibody, a bispecific antibody, a chimeric antibody, a human antibody, a humanized antibody, and an antibody fragment.

280 . The antibody-drug conjugate of claim 265 wherein the antibody fragment is a Fab fragment.

281 . The antibody-drug conjugate of claim 265 wherein the auristatin is MMAE or MMAF.

282 . The antibody-drug conjugate of claim 268 wherein L is 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-val-cit-PAB) or 6-maleimidocaproyl (MC).

283 . An antibody-drug conjugate according to claim 259 having a structure selected from:

wherein Val is valine; Cit is citrulline; p is 1, 2, 3, or 4;

wherein Val is valine; Cit is citrulline; p is 1, 2, 3, or 4;

wherein Val is valine; Cit is citrulline; p is 1, 2, 3, or 4;

or

wherein Val is valine; Cit is citrulline; p is 1, 2, 3, or 4.

284 . The antibody-drug conjugate of claim 283 having the structure

wherein Val is valine; Cit is citrulline; p is 1, 2, 3, or 4.

285 . The antibody-drug conjugate of claim 283 having the structure

wherein Val is valine; Cit is citrulline; p is 1, 2, 3, or 4.

286 . A pharmaceutical formulation for treating cancer comprising an effective amount of the antibody drug conjugate according to of claim 259 and a pharmaceutically acceptable diluent, carrier or excipient.

287 . A method of detecting the presence of a CD79b protein in a sample suspected of containing said protein, said method comprising exposing said sample to an antibody drug conjugate of claim 259 and determining binding of said antibody to said CD79b protein in said sample, wherein said antibody is covalently attached to a label, and wherein binding of the antibody to said protein is indicative of the presence of said protein in said sample.

288 . An assay carried out on a biological sample for detecting cancer cells comprising:

(a) exposing cells to an antibody-drug conjugate of claim 259 ; and

(b) detecting the extent of binding of the antibody-drug conjugate to the cells; wherein the cancer cells are hematopoietic tumor cells.

289 . A method of inhibiting cellular proliferation comprising treating mammalian tumor cells with an antibody-drug conjugate of claim 259 , 283 , 284 or 285 , whereby proliferation of the tumor cells is inhibited.

290 . The method of claim 289 , wherein the mammalian tumor cells are hematopoietic tumor cells.

291 . A method for making an antibody drug conjugate comprising a cysteine engineered anti-CD79b antibody (Ab) according to claim 259 , and an auristatin drug moiety (D) wherein the cysteine engineered antibody is attached through at least one engineered cysteine amino acid by a linker moiety (L) to D; the compound having Formula I:

A b -(L-D) p   I

where p is 1, 2, 3, or 4; the method comprising the steps of:

(a) reacting an engineered cysteine group of the cysteine engineered antibody with a linker reagent to form antibody-linker intermediate Ab-L; and

(b) reacting Ab-L with an activated drug moiety D; whereby the antibody-drug conjugate is formed;

or comprising the steps of:

(c) reacting a nucleophilic group of a drug moiety with a linker reagent to form drug-linker intermediate D-L; and

(d) reacting D-L with an engineered cysteine group of the cysteine engineered antibody;

whereby the antibody-drug conjugate is formed;

wherein the method optionally comprises the step of expressing the cysteine engineered antibody in chinese hamster ovary (CHO) cells.

292 . A method of inhibiting the growth of a cell that expresses CD79b, said method comprising contacting said cell with an antibody-drug conjugate of claim 259 .

293 . A method of treating a proliferative disorder in a subject, said method comprising administering to the subject an effective amount of an antibody-drug conjugate of claim 259 .