Office Action 6: Notice of Allowance (2025-02-12)

CLAIMS:
Listing of Claims:

Amdt date November 21, 2024 Reply to Office action of August 28, 2024 Amendments to the Claims:

This listing of claims will replace all prior versions, and listings, of claims in the application:

1 - 22. (canceled)221

23\. (Previously Presented) A preparation method of an antibody-drug conjugate with a structure as shown in formula I:
mAb-(L-D)ni wherein,
mAb represents a recombinant anti-CD20 monoclonal antibody, wherein the recombinant anti-CD20 monoclonal antibody is rituximab or a biosimilar thereof;
D represents a small molecule toxin, wherein the small molecule toxin is monomethyl auristatin-E (MMAE);
L is a linker connecting the antibody and the small molecule toxin, wherein the linker is maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl;
n is an average number of the small molecule toxins conjugated to the antibody, and n is in a range of 4.2+0.5; and
"-" is a bond, and
wherein the structure of the antibody-drug conjugate is the following formula:mAbSVG {EMBED HERE} 1.2875.817,and wherein the preparation method comprises the steps of:

(1) performing a reduction reaction between the recombinant anti-CD20 monoclonal antibody and a reducing agent in a PB (phosphate buffer) reaction buffer (pH 7.6) at 25±10C for 90±10 minutes, thereby obtaining a reaction system comprising a reduced recombinant anti-CD20 monoclonal antibody, wherein the reducing agent is tris (2-carboxyethyl) phosphine, and a molar ratio of the recombinant anti-CD20 monoclonal antibody to the reducing agent is 1: 2.9 to 1: 3.1;
(2) adding a solution of maleimidocaproyl-vc-PAB-MMAE in acetonitrile and water dropwise to the reaction system of step (1) and performing a coupling reaction at 4±0.5 0C for 60±10 minutes to obtain a coupling solution, wherein a volume ratio of acetonitrile to water is 1:1, and a molar ratio of the recombinant anti-CD20 monoclonal antibody in step (1) to the small molecule toxin is 1: 7.0 to 1: 8.0;
(3) adding into the reaction system obtained from step (2) a stopping solution of L-cysteine which is prepared by dissolving L-cysteine in a diethylenetriaminepentaacetic acid solution, to terminate the coupling reaction; and
(4) when the coupling reaction in step (3) is terminated, transferring the coupling solution to a tangential flow filtration (TFF) system for buffer replacement, thereby replacing the PB reaction buffer with a HT preparation buffer, and filtering the solution obtained with a 0.22 micrometer Rapid - Flow device to make a residual rate of the PB reaction buffer in a final solution less than 0.25%, wherein the HT preparation buffer contains 3.18 g of L-histidine, 70 g of trehalose dihydrate, and 0.2 mL of Tween 80 per L, with pH adjusted to 6.5 using 0.5 mol/L hydrochloric acid, and then filtered through a 0.22 pm membrane.

24\. (Previously Presented) The preparation method of claim 23, wherein n is in a range of 4.2 ±0.3.

25\. (Previously Presented) The preparation method of claim 23, wherein in step (1), the molar ratio of the recombinant anti-CD20 monoclonal antibody to the reducing agent is about 1:2.95-1:3.05.

26\. (Previously Presented) The preparation method of claim 23, wherein in step (1), the molar ratio of the recombinant anti-CD20 monoclonal antibody to the reducing agent is about 1:2.99-1:3.01.

27\. (Previously Presented) The preparation method of claim 23, wherein in step (1), the molar ratio of the recombinant anti-CD20 monoclonal antibody to the reducing agent is about 1:3.

28\. (Previously Presented) The preparation method of claim 23, wherein in step (2), the molar ratio of the recombinant anti-CD20 monoclonal antibody from step (1) to the small molecule toxin is about 1:7.2-1:7.7.

29\. (Previously Presented) The preparation method of claim 23, wherein in step (2), the molar ratio of the recombinant anti-CD20 monoclonal antibody from step (1) to the small molecule toxin is about 1:7.4-1:7.6.

30\. (Previously Presented) The preparation method of claim 23, wherein in step (2), the molar ratio of the recombinant anti-CD20 monoclonal antibody from step (1) to the small molecule toxin is about 1:7.5.

31\. (Previously Presented) The preparation method of claim 23, wherein n is 4.2, and in step (1), the reduction reaction between the recombinant anti-CD20 monoclonal antibody and the reducing agent is performed in the PB reaction buffer (pH

Amdt date November 21, 2024 Reply to Office action of August 28, 2024 7.6) at 25 0C for 90 minutes, the molar ratio of the recombinant anti-CD20 monoclonal antibody to the reducing agent is 1:3; andstep (2), the solution of the small molecule toxin in acetonitrile and water is dropwise added to the reaction system of step (1) and the coupling reaction is performed at 4 0C for 60 minutes, and the molar ratio of the recombinant anti-CD20 monoclonal antibody in step (1) to the small molecule toxin is 1: 7.5.- 35. (Canceled)

0 32

OFFICE ACTION:
DETAILED ACTION
Notice of Pre-AIA  or AIA  Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
The following is an examiner’s statement of reasons for allowance:
The rejection under section 103 is moot since the rejected claims have been canceled.
The rejection under section 112(b) is moot since the rejected claims have been canceled.
Any comments considered necessary by applicant must be submitted no later than the payment of the issue fee and, to avoid processing delays, should preferably accompany the issue fee.  Such submissions should be clearly labeled “Comments on Statement of Reasons for Allowance.”
Conclusion

APPLICANT REBUTTAL: REMARKS/ARGUMENTS
Claims 23-31 are currently pending in the present application. Claims 1-22 were previously canceled, without prejudice and without disclaimer. Claims 32-35 are canceled herein, without prejudice and without disclaimer. No new matter is added. In view of the amendments and remarks that follow, Applicant respectfully requests a timely issuance of a Notice of Allowance of allowed claims 23-31.
Allowable Sub
j
ect Matter
Applicant wishes to thank the Examiner for indicating that claims 23-31 are allowed.
Rej
ection of Claims Under 35 U.S.C.
§
Claims 32-35 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention. In particular, the Office action states that "The criteria of ranking is not clear. Applicant should amend the claims to clearly recite the intended criteria, e.g., amount or concentration of conjugates with the recited ratios.." See OA, page 16.
Without conceding to the propriety of the above definiteness rejection,
in an effort to ex
p
edite allowance
of the present application, Applicant has cancelled claims 32-35, without prejudice and without disclaimer, and thus respectfully submits that the rejection under 35 U.S.C. 112 is now moot and should be withdrawn.
Rej
ection of Claims Under 35 U.S.C.
§
Claims 32-35 were rejected under 35 U.S.C. 103 as allegedly being unpatentable over Law et al., Clin Cancer Res (2004) 10 (23): 7842-7851 (Law); and Francisco et al., Blood. 2003 Aug 15;102(4):1458-65 (Francisco).
In an effort to ex
p
edite allowance
of the present application, and without conceding to this rejection, Applicant has cancelled claims 32-35, without prejudice and without disclaimer, and thus respectfully submits that this rejection under 35 U.S.C. 103 is now moot and should be withdrawn.
For at least the reasons detailed above, it is believed that all rejections are addressed. Thus, Applicant respectfully requests a timely issuance of a Notice of Allowance of allowed claims 23-31.
Concluding Remarks
In view of the foregoing amendments and remarks, Applicant earnestly solicits a timely issuance of a Notice of Allowance. If there are any remaining issues that can be addressed over the telephone, the Examiner is cordially invited to call Applicant's attorney at the number listed below.
Payment of any fee required to enter and process this amendment is provided. The Director is authorized to charge any underpayment of fees or credit any overpayment of fees to Deposit Account No. 03-1728.
Respectfully submitted, LEWIS ROCA ROTHGERBER CHRISTIE LLP


CITED REFERENCE SUMMARIES: Ref US20050180972A1: Claims:
An anti-CD20 antibody-cytotoxic agent conjugate, wherein the cytotoxic agent of the anti-CD20 antibody-cytotoxic agent conjugate has an IC 50 of between 40-fold and 4,000-fold less than the IC 50 of doxorubicin, with the proviso that the cytotoxic agent is not a radioisotope or a toxin, and wherein the IC 50 of each of the cytotoxic agent and doxorubicin is measured by a method comprising: (a) culturing one or more CD20-expressing cell populations in the presence of one or more concentrations of the cytotoxic agent for a 72-to 96-hour period; (b) culturing one or more CD20-expressing cell populations in the presence of one or more concentrations of doxorubicin for a 72- to 96-hour period; and (c) identifying a concentration of the cytotoxic agent and doxorubicin, respectively, at which 50% fewer cells in the CD20-expressing cell populations of steps (a) and (b), respectively, are viable at the end of the period relative to a CD20-expressing cell population cultured in the absence of the cytotoxic agent and doxorubicin, wherein the CD20-expressing cell populations of steps (a), (b) and (c) are of the same cell type and are cultured under the same conditions, and wherein the concentration of the cytotoxic agent and doxorubicin identified in step (c) is the IC 50 of the cytotoxic agent and doxorubicin, respectively.
(a) culturing one or more CD20-expressing cell populations in the presence of one or more concentrations of the cytotoxic agent for a 72-to 96-hour period;
(b) culturing one or more CD20-expressing cell populations in the presence of one or more concentrations of doxorubicin for a 72- to 96-hour period; and
(c) identifying a concentration of the cytotoxic agent and doxorubicin, respectively, at which 50% fewer cells in the CD20-expressing cell populations of steps (a) and (b), respectively, are viable at the end of the period relative to a CD20-expressing cell population cultured in the absence of the cytotoxic agent and doxorubicin,
wherein the CD20-expressing cell populations of steps (a), (b) and (c) are of the same cell type and are cultured under the same conditions,
and wherein the concentration of the cytotoxic agent and doxorubicin identified in step (c) is the IC 50 of the cytotoxic agent and doxorubicin, respectively.
- 5 . (canceled)
An anti-CD20 antibody-cytotoxic agent conjugate, wherein the anti-CD20 antibody-cytotoxic agent conjugate has an IC 50 of between 40-fold and 4,000-fold less than the IC 50 of an anti-CD20 antibody-doxorubicin conjugate, with the proviso that the cytotoxic agent is not a radioisotope or a toxin, wherein the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate comprise the same anti-CD20 antibody, and wherein the IC 50 of each of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate is measured by a method comprising: (a) culturing one or more CD20-expressing cell populations in the presence of one or more concentrations of the anti-CD20 antibody-cytotoxic agent conjugate for a 72- to 96-hour period; (b) culturing one or more CD20-expressing cell populations in the presence of one or more concentrations of the anti-CD20 antibody-doxorubicin conjugate for a 72- to 96-hour period; and (c) identifying a concentration of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate, respectively, at which 50% fewer cells in the CD20-expressing cell populations of steps (a) and (b), respectively, are viable at the end of the period relative to a CD20-expressing cell population type cultured in the absence of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate, wherein the CD20-expressing cell populations of steps (a), (b) and (c) are of the same cell type and are cultured under the same conditions, and wherein the concentration of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate identified in step (c) is the IC 50 of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate, respectively.
(a) culturing one or more CD20-expressing cell populations in the presence of one or more concentrations of the anti-CD20 antibody-cytotoxic agent conjugate for a 72- to 96-hour period;
(b) culturing one or more CD20-expressing cell populations in the presence of one or more concentrations of the anti-CD20 antibody-doxorubicin conjugate for a 72- to 96-hour period; and
(c) identifying a concentration of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate, respectively, at which 50% fewer cells in the CD20-expressing cell populations of steps (a) and (b), respectively, are viable at the end of the period relative to a CD20-expressing cell population type cultured in the absence of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate,
and wherein the concentration of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate identified in step (c) is the IC 50 of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate, respectively.
- 11 . (canceled)
An anti-CD20 antibody-cytotoxic agent conjugate, wherein the conjugate has a rate of accumulation in a CD20-expressing cell that is between 20-fold and 5,000-fold greater than the rate of accumulation of an unconjugated form of the anti-CD20 antibody in the CD20-expressing cell, wherein the rates of accumulation of the conjugate and of the unconjugated form of the antibody are measured by a method comprising: (a) culturing a population of the CD20-expressing cell with the conjugate; (b) culturing a population of the CD20-expressing cell with the unconjugated antibody, wherein the populations of steps (a) and (b) are cultured under the same conditions; and (c) measuring the amount of the conjugate and unconjugated antibody accumulated in the populations of steps (a) and (b), respectively.
(a) culturing a population of the CD20-expressing cell with the conjugate;
(b) culturing a population of the CD20-expressing cell with the unconjugated antibody, wherein the populations of steps (a) and (b) are cultured under the same conditions; and
(c) measuring the amount of the conjugate and unconjugated antibody accumulated in the populations of steps (a) and (b), respectively.
- 17 . (canceled)
An anti-CD20 antibody-cytotoxic agent conjugate, wherein the conjugate has a rate of accumulation in a CD20-expressing cell that is between 20-fold and 5,000-fold greater than the rate of accumulation of an anti-CD20 antibody-doxorubicin conjugate in a CD20-expressing cell of the same cell type, wherein the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate comprise the same anti-CD20 antibody, and wherein the rates of accumulation of the anti-CD20 antibody-cytotoxic agent conjugate and of the anti-CD20 antibody-doxorubicin conjugate are measured by a method comprising: (a) culturing a population of the CD20-expressing cell with the anti-CD20 antibody-cytotoxic agent conjugate; (b) culturing a population of the CD20-expressing cell with the anti-CD20 antibody-doxorubicin conjugate, wherein the populations of steps (a) and (b) are cultured under the same conditions; and (c) measuring the amount of the anti-CD20 antibody-cytotoxic agent conjugate and anti-CD20 antibody-doxorubicin conjugate accumulated in the populations of steps (a) and (b), respectively.
(a) culturing a population of the CD20-expressing cell with the anti-CD20 antibody-cytotoxic agent conjugate;
(b) culturing a population of the CD20-expressing cell with the anti-CD20 antibody-doxorubicin conjugate, wherein the populations of steps (a) and (b) are cultured under the same conditions; and
(c) measuring the amount of the anti-CD20 antibody-cytotoxic agent conjugate and anti-CD20 antibody-doxorubicin conjugate accumulated in the populations of steps (a) and (b), respectively.
- 21 . (canceled)
An anti-CD20 antibody-cytotoxic agent conjugate, wherein the conjugate exhibits an accumulation in a non-peripheral region inside a CD20-expressing cell that is between 1.5-fold and 5,000-fold greater than the accumulation of an unconjugated form of the anti-CD20 antibody in the CD20-expressing cell, wherein the accumulation of the conjugate and of the unconjugated form of the antibody are measured by a method comprising: (a) culturing a population of the CD20-expressing cell with the conjugate; (b) culturing a population of the CD20-expressing cell with the unconjugated form of the anti-CD20 antibody; and (c) detecting by confocal fluorescence microscopy localization of the conjugate and the unconjugated form of the anti-CD20 antibody in the populations of steps (a) and (b), respectively, wherein the populations of steps (a) and (b) are cultured under the same conditions and for the same period of time, and wherein the conjugate exhibits an accumulation in the CD20-expressing cell that is between 1.5-fold and 5,000-fold greater than the accumulation of the unconjugated form of the anti-CD20 antibody in the CD20-expressing cell if: (i) between 1.5-fold and 5,000-fold as many cells of the population of step (a) contain a detectable amount of the conjugate in a non-peripheral region as the number of cells of the population of step (b) contain the unconjugated form of the antibody in a non-peripheral region; or (ii) the accumulation of the conjugate in a non-peripheral region of the majority of CD20-expressing cells of the population of step (a) is between 1.5-fold and 5,000-fold greater than the accumulation of the unconjugated form of the anti-CD20 antibody in the majority of CD20-expressing cells of the population of step (b).
(b) culturing a population of the CD20-expressing cell with the unconjugated form of the anti-CD20 antibody; and
(c) detecting by confocal fluorescence microscopy localization of the conjugate and the unconjugated form of the anti-CD20 antibody in the populations of steps (a) and (b), respectively,
wherein the populations of steps (a) and (b) are cultured under the same conditions and for the same period of time, and wherein the conjugate exhibits an accumulation in the CD20-expressing cell that is between 1.5-fold and 5,000-fold greater than the accumulation of the unconjugated form of the anti-CD20 antibody in the CD20-expressing cell if:
(i) between 1.5-fold and 5,000-fold as many cells of the population of step (a) contain a detectable amount of the conjugate in a non-peripheral region as the number of cells of the population of step (b) contain the unconjugated form of the antibody in a non-peripheral region; or
(ii) the accumulation of the conjugate in a non-peripheral region of the majority of CD20-expressing cells of the population of step (a) is between 1.5-fold and 5,000-fold greater than the accumulation of the unconjugated form of the anti-CD20 antibody in the majority of CD20-expressing cells of the population of step (b).
- 25 . (canceled)
An anti-CD20 antibody-cytotoxic agent conjugate, wherein the conjugate exhibits an accumulation in a non-peripheral region inside a CD20-expressing cell that is between 1.5-fold and 5,000-fold greater than the accumulation of an anti-CD20 antibody-doxorubicin conjugate, wherein the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate comprise the same anti-CD20 antibody, in the CD20-expressing cell, wherein the accumulation of the conjugate and of the anti-CD20 antibody-doxorubicin conjugate are measured by a method comprising: (a) culturing a population of the CD20-expressing cell with the conjugate; (b) culturing a population of the CD20-expressing cell with the anti-CD20 antibody-doxorubicin conjugate; and (c) detecting by confocal fluorescence microscopy localization of the conjugate and the anti-CD20 antibody-doxorubicin conjugate in the populations of steps (a) and (b), respectively, wherein the populations of steps (a) and (b) are cultured under the same conditions and for the same period of time, and wherein the conjugate exhibits an accumulation in the CD20-expressing cell that is between 1.5-fold and 5,000-fold greater than the accumulation of the anti-CD20 antibody-doxorubicin conjugate in the CD20-expressing cell if: (i) between 1.5-fold and 5,000-fold as many cells of the population of step (a) contain a detectable amount of the conjugate in a non-peripheral region as the number of cells of the population of step (b) contain the anti-CD20 antibody-doxorubicin conjugate in a non-peripheral region; or (ii) the accumulation of the conjugate in a non-peripheral region of the majority of CD20-expressing cells of the population of step (a) is between 1.5-fold and 5,000-fold greater than the accumulation of the anti-CD20 antibody-doxorubicin conjugate in the majority of CD20-expressing cells of the population of step (b).
(b) culturing a population of the CD20-expressing cell with the anti-CD20 antibody-doxorubicin conjugate; and
(c) detecting by confocal fluorescence microscopy localization of the conjugate and the anti-CD20 antibody-doxorubicin conjugate in the populations of steps (a) and (b), respectively,
wherein the populations of steps (a) and (b) are cultured under the same conditions and for the same period of time, and wherein the conjugate exhibits an accumulation in the CD20-expressing cell that is between 1.5-fold and 5,000-fold greater than the accumulation of the anti-CD20 antibody-doxorubicin conjugate in the CD20-expressing cell if:
(i) between 1.5-fold and 5,000-fold as many cells of the population of step (a) contain a detectable amount of the conjugate in a non-peripheral region as the number of cells of the population of step (b) contain the anti-CD20 antibody-doxorubicin conjugate in a non-peripheral region; or
(ii) the accumulation of the conjugate in a non-peripheral region of the majority of CD20-expressing cells of the population of step (a) is between 1.5-fold and 5,000-fold greater than the accumulation of the anti-CD20 antibody-doxorubicin conjugate in the majority of CD20-expressing cells of the population of step (b).
- 64 . (canceled)
A pharmaceutical composition comprising an anti-CD20 antibody-cytotoxic agent conjugate, wherein the cytotoxic agent of the anti-CD20 antibody-cytotoxic agent conjugate has an IC 50 of between 40-fold and 4,000-fold less than the IC 50 of doxorubicin, with the proviso that the cytotoxic agent is not a radioisotope or a toxin, and wherein the IC 50 of each of the cytotoxic agent and doxorubicin is measured by a method comprising: (a) culturing one or more CD20-expressing cell populations in the presence of one or more concentrations of the cytotoxic agent for a 72-to 96-hour period; (b) culturing one or more CD20-expressing cell populations in the presence of one or more concentrations of doxorubicin for a 72- to 96-hour period; and (c) identifying a concentration of the cytotoxic agent and doxorubicin, respectively, at which 50% fewer cells in the CD20-expressing cell populations of steps (a) and (b), respectively, are viable at the end of the period relative to a CD20-expressing cell population cultured in the absence of the cytotoxic agent and doxorubicin, wherein the CD20-expressing cell populations of steps (a), (b) and (c) are of the same cell type and are cultured under the same conditions, and wherein the concentration of the cytotoxic agent and doxorubicin identified in step (c) is the IC 50 of the cytotoxic agent and doxorubicin, respectively.
A pharmaceutical composition comprising an anti-CD20 antibody-cytotoxic agent conjugate, wherein the anti-CD20 antibody-cytotoxic agent conjugate has an IC 50 of between 40-fold and 4,000-fold less than the IC 50 of an anti-CD20 antibody-doxorubicin conjugate, with the proviso that the cytotoxic agent is not a radioisotope or a toxin, wherein the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate comprise the same anti-CD20 antibody, and wherein the IC 50 of each of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate is measured by a method comprising: (a) culturing one or more CD20-expressing cell populations in the presence of one or more concentrations of the anti-CD20 antibody-cytotoxic agent conjugate for a 72- to 96-hour period; (b) culturing one or more CD20-expressing cell populations in the presence of one or more concentrations of the anti-CD20 antibody-doxorubicin conjugate for a 72- to 96-hour period; and (c) identifying a concentration of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate, respectively, at which 50% fewer cells in the CD20-expressing cell populations of steps (a) and (b), respectively, are viable at the end of the period relative to a CD20-expressing cell population cultured in the absence of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate, wherein the CD20-expressing cell populations of steps (a), (b) and (c) are of the same cell type and are cultured under the same conditions, and wherein the concentration of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate identified in step (c) is the IC 50 of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate, respectively.
(c) identifying a concentration of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate, respectively, at which 50% fewer cells in the CD20-expressing cell populations of steps (a) and (b), respectively, are viable at the end of the period relative to a CD20-expressing cell population cultured in the absence of the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate,
A pharmaceutical composition comprising an anti-CD20 antibody-cytotoxic agent conjugate, wherein the conjugate has a rate of accumulation in a CD20-expressing cell that is between 20-fold and 5,000-fold greater than the rate of accumulation of an unconjugated form of the anti-CD20 antibody in the CD20-expressing cell, with the proviso that the cytotoxic agent is not a radioisotope or a toxin, wherein the rates of accumulation of the conjugate and of the unconjugated form of the antibody are measured by a method comprising: (a) culturing a population of the CD20-expressing cell with the conjugate; (b) culturing a population of the CD20-expressing cell with the unconjugated antibody, wherein the populations of steps (a) and (b) are cultured under the same conditions; and (c) measuring the amount of the conjugate and unconjugated antibody accumulated in the populations of steps (a) and (b), respectively.
A pharmaceutical composition comprising an anti-CD20 antibody-cytotoxic agent conjugate, wherein the conjugate has a rate of accumulation in a CD20-expressing cell that is between 20-fold and 5,000-fold greater than the rate of accumulation of an anti-CD20 antibody-doxorubicin conjugate in a CD20-expressing cell of the same cell type, with the proviso that the cytotoxic agent is not a radioisotope or a toxin, wherein the anti-CD20 antibody-cytotoxic agent conjugate and the anti-CD20 antibody-doxorubicin conjugate comprise the same anti-CD20 antibody, and wherein the rates of accumulation of the anti-CD20 antibody-cytotoxic agent conjugate and of the anti-CD20 antibody-doxorubicin conjugate are measured by a method comprising: (a) culturing a population of the CD20-expressing cell with the anti-CD20 antibody-cytotoxic agent conjugate; (b) culturing a population of the CD20-expressing cell with the anti-CD20 antibody-doxorubicin conjugate, wherein the populations of steps (a) and (b) a
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