METHODS FOR INCREASING EFFICACY OF FOLR1 CANCER THERAPY

1 .- 19 . (canceled)

20 . An article of manufacture comprising an anti-FOLR1 antibody or an anti-FOLR1 immunoconjugate; a container; and a package insert or label indicating that the antibody or immunoconjugate can be used to treat a cancer characterized by the expression of FOLR1 at a level of 2, 3, or 3+ measured by IHC.

21 .- 96 . (canceled)

97 . A method for treating ovarian cancer comprising administering a therapeutically effective dose of an anti-Folate Receptor 1 (FOLR1) immunoconjugate to a human subject having ovarian cancer; wherein a FOLR1 staining intensity score of 2 or greater has been detected in greater than 25% of the cells in a tumor sample from the human subject using an immunohistochemistry (IHC) detection method that distinguishes between staining intensity and staining uniformity.

98 . The method of claim 97 , wherein the anti-FOLR1 immunoconjugate has the formula (A)-(L)-(C), wherein:

(A) comprises an antibody or antigen binding fragment thereof comprising: (a) a heavy chain CDR1 comprising the amino acid sequence GYFMN (SEQ ID NO:6); a heavy chain CDR2 comprising the amino acid sequence RIHPYDGDTFYNQKFQG (SEQ ID NO:7); and a heavy chain CDR3 comprising the amino acid sequence YDGSRAMDY (SEQ ID NO:8); and (b) a light chain CDR1 comprising the amino acid sequence KASQSVSFAGTSLMH (SEQ ID NO:9); a light chain CDR2 comprising the amino acid sequence RASNLEA (SEQ ID NO:10); and a light chain CDR3 comprising the amino acid sequence QQSREYPYT (SEQ ID NO:11),

(L) comprises the linker N-succinimidyl 4-(2-pyridyldithio)-2-sulfobutanoate (sulfo-SPDB), and

(C) comprises the cytotoxic agent N(2′)-deacetyl-N(2′)-(4-mercapto-4-methyl-1-oxopentyl)-maytansine (DM4), and

wherein the linker (L) links (A) to (C).

99 . The method of claim 99 , wherein the antibody or antigen binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 5.

100 . The method of claim 99 , wherein the antibody comprises (i) a heavy chain comprising the same amino acid sequence as the amino acid sequence of the heavy chain encoded by the plasmid deposited with the American Type Culture Collection (ATCC) as PTA-10772 and (ii) a light chain comprising the same amino acid sequence as the amino acid sequence of the light chain encoded by the plasmid deposited with the ATCC as PTA-10774.

101 . The method of claim 97 , wherein a FOLR1 staining intensity score of 2 or greater has been detected in greater than 75% of the cells in a tumor sample from the human subject using the IHC detection method.

102 . The method of claim 101 , wherein:

(A) comprises an antibody or antigen binding fragment thereof comprising: a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 5,

(L) comprises the linker N-succinimidyl 4-(2-pyridyldithio)-2-sulfobutanoate (sulfo-SPDB), and

(C) comprises the cytotoxic agent N(2′)-deacetyl-N(2′)-(4-mercapto-4-methyl-1-oxopentyl)-maytansine (DM4), and

wherein the linker (L) links (A) to (C).

103 . The method of claim 102 , wherein the antibody comprises (i) a heavy chain comprising the same amino acid sequence as the amino acid sequence of the heavy chain encoded by the plasmid deposited with the American Type Culture Collection (ATCC) as PTA-10772 and (ii) a light chain comprising the same amino acid sequence as the amino acid sequence of the light chain encoded by the plasmid deposited with the ATCC as PTA-10774.

104 . The method of claim 97 , further comprising detecting FOLR1 expression in the tumor sample from the human subject using the IHC detection method prior to administering the therapeutically effective dose of the anti-FOLR1 immunoconjugate to the human subject.

105 . The method of claim 97 , wherein the tumor sample is a formalin fixed paraffin embedded sample.

106 . The method of claim 97 , wherein the IHC detection method comprises detecting FOLR1 expression with a detection antibody or antigen binding fragment thereof that specifically binds FOLR1, wherein the detection antibody or antigen binding fragment thereof comprises a detection reagent selected from the group consisting of: an enzyme, a fluorophore, a radioactive label, and a luminophore.

107 . The method of claim 106 , wherein the detection reagent is selected from the group consisting of: biotin, digoxigenin, fluorescein, tritium, and rhodamine.

108 . The method of claim 97 , wherein a staining intensity score of 3 or greater for FOLR1 expression has been detected in greater than 75% of cells of the tumor sample.

109 . A method for treating ovarian cancer comprising:

(a) contacting a tumor tissue sample from a human subject having ovarian cancer with a detection antibody or antigen binding fragment thereof that specifically binds FOLR1, wherein the sample is formalin-fixed paraffin embedded;

(b) measuring the binding of the detection antibody or antigen binding fragment thereof to FOLR1 in the tumor tissue sample in step (a) using an IHC detection method that can distinguish between staining intensity and staining uniformity in the FOLR1 expressing tumor tissue sample as compared to staining intensity or staining uniformity in one or more reference samples;

(c) assigning a FOLR1 expression score to the tumor tissue sample after comparing the level of FOLR1 staining intensity and staining uniformity in the tumor tissue sample to one or more reference samples; and

(d) administering an anti-FOLR1 immunoconjugate to the human subject whose tumor tissue sample has greater than 25% of cells having a FOLR1 staining intensity score of 2 or greater.

110 . The method of claim 109 , wherein the tumor tissue sample has greater than 75% of cells having a FOLR1 staining intensity score of 2 or greater.

111 . The method of claim 109 , wherein the anti-FOLR1 immunoconjugate has the formula (A)-(L)-(C), wherein:

(A) comprises an antibody or antigen binding fragment thereof comprising: (a) a heavy chain CDR1 comprising the amino acid sequence GYFMN (SEQ ID NO:6); a heavy chain CDR2 comprising the amino acid sequence RIHPYDGDTFYNQKFQG (SEQ ID NO:7); and a heavy chain CDR3 comprising the amino acid sequence YDGSRAMDY (SEQ ID NO:8); and (b) a light chain CDR1 comprising the amino acid sequence KASQSVSFAGTSLMH (SEQ ID NO:9); a light chain CDR2 comprising the amino acid sequence RASNLEA (SEQ ID NO:10); and a light chain CDR3 comprising the amino acid sequence QQSREYPYT (SEQ ID NO:11),

(L) comprises the linker N-succinimidyl 4-(2-pyridyldithio)-2-sulfobutanoate (sulfo-SPDB), and

(C) comprises the cytotoxic agent N(2′)-deacetyl-N(2′)-(4-mercapto-4-methyl-1-oxopentyl)-maytansine (DM4), and

wherein the linker (L) links (A) to (C).

112 . The method of claim 111 , wherein the antibody or antigen binding fragment thereof of the immunoconjugate comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 5.

113 . The method of claim 111 , wherein the antibody of the immunoconjugate comprises (i) a heavy chain comprising the same amino acid sequence as the amino acid sequence of the heavy chain encoded by the plasmid deposited with the ATCC as PTA-10772 and (ii) a light chain comprising the same amino acid sequence as the amino acid sequence of the light chain encoded by the plasmid deposited with the ATCC as PTA-10774.

114 . The method of claim 109 , wherein the tumor tissue sample is a formalin fixed paraffin embedded sample.

115 . The method of claim 109 , wherein the tumor tissue sample has greater than 75% of cells having a staining intensity score of 3 or greater.