JP2020519640A - Anti-CD33 immunoconjugate administration plan - Google Patents

Abstract

The invention provides a method of treating cancer in a subject, comprising administering to the subject an anti-CD33 immunoconjugate (eg, IMGN779) at a therapeutically effective dose. The present invention has discovered that, at least in part, specific dosing regimens of anti-CD33 immunoconjugates (eg, IMGN779) are useful in the treatment of patients suffering from cancer, particularly blood cancers such as AML. Regarding Thus, provided herein are methods of treating cancer with anti-CD33 immunoconjugates (eg, IMGN779). [Selection diagram] None

Description

Treatment of hematological malignancies with conventional chemotherapy is often associated with severe morbidity because chemotherapeutic agents are not specific to cancerous cells. Moreover, despite a high initial response rate to chemotherapy, many patients with hematological malignancies (eg, acute myeloid leukemia) fail to achieve complete remission and relapse within a relatively short period of time after diagnosis.

The leukocyte differentiation antigen CD33 is a 364 amino acid transmembrane glycoprotein with sequence homology to members of the sialoadhesin family including glycoproteins related to myelin and CD22, and sialoadhesin itself (S. Peiper, 2002). , Leucoty Typing VII, White Cell Differentiation, Antigens, Proceedings of the Seventh International Workshop and Conference, Oxford University, 77.).

Expression of CD33 appears to be highly specific for hematopoietic compartments, with strong expression by myeloid progenitor cells (S. Peiper, 2002). It has reduced expression upon maturation and differentiation, but myeloid progenitor cells such as CFU-GEMM, CFU-GM, CFU-G and BFU-E, granulocyte precursors such as monocytes/macrophages, promyelocytes and myelocytes. It is expressed in the body as well as by mature granulocytes at low expression levels (S. Peiper, 2002). Anti-CD33 monoclonal antibodies have shown that clonogenic acute myeloid leukemia (AML) cells express CD33 in more than 80% of human cases (LaRussa, VF et al., 1992, Exp. Hematol. 20:442-448). In contrast, pluripotent hematopoietic stem cells (Leary, AG et al., 1987, Blood 69:953) that give rise to "blast colonies" in vitro and pluripotent to induce hematopoietic long-term marrow cultures. Sexual hematopoietic stem cells (Andrews RG et al., 1989, J. Exp. Med. 169:1721; Sutherland, HJ et al., 1989, Blood 74:1563) appear to lack CD33 expression. Looks like.

Immunoconjugates (also known as antibody drug conjugates or ADCs) that combine a cytotoxic agent with a monoclonal antibody that specifically recognizes and binds to CD33 by selectively expressing CD33 are associated with hematological malignancies. Use in the selective targeting of tumors (eg AML cells) has been proposed. However, after 10 years on the market, the anti-CD33 immunoconjugate Mylotarg® (gemtuzumab ozogamicin) was voluntarily collected from the market as a result of safety and efficacy concerns. As such, there remains a need to identify specific dosing regimens for specific anti-CD33 immunoconjugates for treating patients suffering from cancer, particularly blood cancers such as AML.

The present invention has discovered that, at least in part, specific dosing regimens of anti-CD33 immunoconjugates (eg, IMGN779) are useful in the treatment of patients suffering from cancer, particularly blood cancers such as AML. Regarding Thus, provided herein are methods of treating cancer with anti-CD33 immunoconjugates (eg, IMGN779).

In one embodiment, the method of treating cancer in a subject comprises:
The following formula:

Comprising administering to the subject about 1 to about 2 mg/kg of an anti-CD33 immunoconjugate of or a pharmaceutically acceptable salt thereof, wherein the doublet between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a is variable heavy chain comprising the CDRl, CDR2, and CDR3 sequences set forth in SEQ ID NO: 1-3, and SEQ ID NO: 4-6 An anti-CD33 antibody or an antigen-binding fragment thereof, which comprises a variable light chain comprising the CDR1, CDR2, and CDR3 sequences described, respectively.

In one embodiment, the method of treating cancer in a subject comprises:
The following formula:

Comprising administering to the subject about 0.70 to about 1.25 mg/kg of the anti-CD33 immunoconjugate or a pharmaceutically acceptable salt thereof, wherein the doublet between N and C:

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a is variable heavy chain comprising the CDRl, CDR2, and CDR3 sequences set forth in SEQ ID NO: 1-3, and SEQ ID NO: 4-6 An anti-CD33 antibody or an antigen-binding fragment thereof, which comprises a variable light chain comprising the CDR1, CDR2, and CDR3 sequences described, respectively.

In one embodiment, the method of treating cancer in a subject comprises:
The following formula:

Comprising administering to the subject about 0.54 to about 1.0 mg/kg of an anti-CD33 immunoconjugate or a pharmaceutically acceptable salt thereof, wherein: a doublet between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a is variable heavy chain comprising the CDRl, CDR2, and CDR3 sequences set forth in SEQ ID NO: 1-3, and SEQ ID NO: 4-6 An anti-CD33 antibody or an antigen-binding fragment thereof, which comprises a variable light chain comprising the CDR1, CDR2, and CDR3 sequences described, respectively.

In one embodiment, the method of treating cancer in a subject comprises:
The following formula:

Comprising administering to the subject about 0.54 to about 0.8 mg/kg of an anti-CD33 immunoconjugate or a pharmaceutically acceptable salt thereof, wherein: the doublet between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a is variable heavy chain comprising the CDRl, CDR2, and CDR3 sequences set forth in SEQ ID NO: 1-3, and SEQ ID NO: 4-6 An anti-CD33 antibody or an antigen-binding fragment thereof, which comprises a variable light chain comprising the CDR1, CDR2, and CDR3 sequences described, respectively.

In one embodiment, about 0.54 to about 0.70 mg/kg of immunoconjugate is administered. In one embodiment, about 0.54 or about 0.70 mg/kg of the immunoconjugate is administered.

In one embodiment, about 0.5 mg/kg of the immunoconjugate is administered. In one embodiment, about 0.6 mg/kg of immunoconjugate is administered. In one embodiment, about 0.7 mg/kg of immunoconjugate is administered. In one embodiment, about 0.75 mg/kg of the immunoconjugate is administered. In one embodiment, about 0.8 mg/kg of immunoconjugate is administered. In one embodiment, about 0.85 mg/kg of immunoconjugate is administered. In one embodiment, about 0.9 mg/kg of the immunoconjugate is administered. In one embodiment, about 0.91 mg/kg of the immunoconjugate is administered. In one embodiment, about 0.92 mg/kg of the immunoconjugate is administered. In one embodiment, about 0.93 mg/kg of immunoconjugate is administered. In one embodiment, about 0.94 mg/kg of immunoconjugate is administered. In one embodiment, about 0.95 mg/kg of immunoconjugate is administered. In one embodiment, about 0.96 mg/kg of the immunoconjugate is administered. In one embodiment, about 0.97 mg/kg of immunoconjugate is administered. In one embodiment, about 0.98 mg/kg of immunoconjugate is administered. In one embodiment, about 0.99 mg/kg of immunoconjugate is administered. In one embodiment, about 1.0 mg/kg of the immunoconjugate is administered.

In one embodiment, about 1.01 mg/kg of immunoconjugate is administered. In one embodiment, about 1.02 mg/kg of immunoconjugate is administered. In one embodiment, about 1.03 mg/kg of immunoconjugate is administered. In one embodiment, about 1.04 mg/kg of immunoconjugate is administered. In one embodiment, about 1.05 mg/kg of immunoconjugate is administered. In one embodiment, about 1.06 mg/kg of immunoconjugate is administered. In one embodiment, about 1.07 mg/kg of immunoconjugate is administered. In one embodiment, about 1.08 mg/kg of immunoconjugate is administered. In one embodiment, about 1.09 mg/kg of immunoconjugate is administered.

In one embodiment, about 1.1 mg/kg of immunoconjugate is administered. In one embodiment, about 1.11 mg/kg of immunoconjugate is administered. In one embodiment, about 1.12 mg/kg of immunoconjugate is administered. In one embodiment, about 1.13 mg/kg of immunoconjugate is administered. In one embodiment, about 1.14 mg/kg of immunoconjugate is administered. In one embodiment, about 1.15 mg/kg of the immunoconjugate is administered. In one embodiment, about 1.16 mg/kg of immunoconjugate is administered. In one embodiment, about 1.17 mg/kg of immunoconjugate is administered. In one embodiment, about 1.18 mg/kg of immunoconjugate is administered. In one embodiment, about 1.19 mg/kg of immunoconjugate is administered.

In one embodiment, about 1.2 mg/kg of immunoconjugate is administered. In one embodiment, about 1.21 mg/kg of immunoconjugate is administered. In one embodiment, about 1.22 mg/kg of immunoconjugate is administered. In one embodiment, about 1.23 mg/kg of the immunoconjugate is administered. In one embodiment, about 1.24 mg/kg of immunoconjugate is administered. In one embodiment, about 1.25 mg/kg of immunoconjugate is administered.

In one embodiment, about 1.5 mg/kg of immunoconjugate is administered. In one embodiment, about 1.51 mg/kg of immunoconjugate is administered. In one embodiment, about 1.52 mg/kg of immunoconjugate is administered. In one embodiment, about 1.53 mg/kg of immunoconjugate is administered. In one embodiment, about 1.54 mg/kg of immunoconjugate is administered. In one embodiment, about 1.55 mg/kg of immunoconjugate is administered. In one embodiment, about 1.56 mg/kg of immunoconjugate is administered. In one embodiment, about 1.57 mg/kg of immunoconjugate is administered. In one embodiment, about 1.58 mg/kg of immunoconjugate is administered. In one embodiment, about 1.59 mg/kg of the immunoconjugate is administered. In one embodiment, about 1.6 mg/kg of the immunoconjugate is administered.

In one embodiment, about 2 mg/kg of immunoconjugate is administered.

In one embodiment, the immunoconjugate is administered about once every two weeks.

In one embodiment, the method of treating cancer in a subject comprises:
The following formula:

Comprising administering to the subject about 0.30 to about 0.54 mg/kg of an anti-CD33 immunoconjugate or a pharmaceutically acceptable salt thereof, wherein: a doublet between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a is variable heavy chain comprising the CDRl, CDR2, and CDR3 sequences set forth in SEQ ID NO: 1-3, and SEQ ID NO: 4-6 An anti-CD33 antibody or an antigen-binding fragment thereof, which comprises a variable light chain comprising the CDR1, CDR2, and CDR3 sequences described, respectively.

In one embodiment, about 0.39 to about 0.54 mg/kg of immunoconjugate is administered. In one embodiment, about 0.39 mg/kg or about 0.54 mg/kg immunoconjugate is administered.

In one embodiment, about 0.45 mg/kg of immunoconjugate is administered. In one embodiment, about 0.5 mg/kg of the immunoconjugate is administered. In one embodiment, about 0.55 mg/kg of immunoconjugate is administered. In one embodiment, about 0.6 mg/kg of immunoconjugate is administered. In one embodiment, about 0.65 mg/kg of immunoconjugate is administered. In one embodiment, about 0.7 mg/kg of immunoconjugate is administered.

In one embodiment, the immunoconjugate is administered about once per week.

In one embodiment, the immunoconjugate is administered on a 28-day cycle.

In one embodiment, the immunoconjugate is administered by intravenous infusion.

In one embodiment, the variable heavy chain comprises the sequence set forth in SEQ ID NO:9 and the variable light chain comprises the sequence set forth in SEQ ID NO:10.

In one embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain comprising the sequence set forth in SEQ ID NO:11 and a light chain comprising the sequence set forth in SEQ ID NO:12.

In one embodiment, the immunoconjugate is
Formula (II):

Or an pharmaceutically acceptable salt thereof, wherein A is an anti-CD33 antibody or an antigen-binding fragment thereof, and r is an integer of 1 to 10.

In one embodiment, the immunoconjugate is
Formula (III):

Or an pharmaceutically acceptable salt thereof, wherein A is an anti-CD33 antibody or an antigen-binding fragment thereof, and r is an integer of 1 to 10.

In one embodiment, the pharmaceutically acceptable salt is the sodium or potassium salt.

In one embodiment, r is an integer from 2-9, 3-8, 4-7, or 5-6.

In one embodiment, the immunoconjugate is included in a composition comprising at least two of the immunoconjugates and the average number of cytotoxins per antibody or antigen binding fragment thereof is 2-8, 3-7. 3 to 5 or 2.5 to 3.5.

In one embodiment, the indolinobenzodiazepine dimer (eg, DGN462) is linked to the anti-CD33 antibody or antigen binding fragment thereof via the lysine residue of the antibody or antigen binding fragment thereof. In one embodiment, 1-10, 2-9, 3-8, 4-7, or 5-6 indolinobenzodiazepine dimer molecules (eg, DGN462) are lysine residues of the antibody or antigen-binding fragment thereof. Via anti-CD33 antibody or an antigen-binding fragment thereof. In one embodiment, 2-8, 2-7, 3-5, or 2.5-3.5 indolinobenzodiazepine dimer molecules (eg, DGN462) are lysine residues of the antibody or antigen-binding fragment thereof. Via anti-CD33 antibody or an antigen-binding fragment thereof.

In one embodiment, the immunoconjugate is IMGN779.

In one embodiment, the cancer is selected from the group consisting of leukemia, lymphoma and myeloma. In one embodiment, the cancer is acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute lymphoblastic leukemia (ALL), B cell lineage of acute lymphoblastic leukemia (B ALL), T cell acute lymphoblastic leukemia (T ALL) chronic lymphocytic leukemia (CLL), hair cell leukemia (HCL), myelodysplastic syndrome (MDS), basic plasmacytoid DC tumor (BPDCN) leukemia, non It is selected from the group consisting of Hodgkin lymphoma (NHL), mantle cell lymphoma, and Hodgkin leukemia (HL). In one embodiment, the cancer is acute myelogenous leukemia (AML).

In one embodiment, AML is refractory or relapsing acute myelogenous leukemia. In one embodiment, AML has overexpression of P-glycoprotein, overexpression of EVI1, p53 mutation, DNMT3A mutation, FLT3 internal tandem duplication, decreased expression of complex karyotype, BRCA1, BRCA2, or PALB2, or BRCA1, Characterized by mutations in BRCA2, or PALB2.

In one embodiment, the homozygous rs12459419C genotype has been detected in a sample obtained from the subject. In one embodiment, the method further comprises detecting the homozygous rs12459419C genotype in a sample obtained from the subject.

In one embodiment, the heterozygous rs12459419C genotype has been detected in a sample obtained from the subject. In one embodiment, the method further comprises detecting the heterozygous rs12459419C genotype in a sample obtained from the subject.

In one embodiment, the homozygous rs12459419T genotype has been detected in a sample obtained from the subject. In one embodiment, the method of treating cancer comprises detecting a homozygous rs12459419T genotype in a sample obtained from a subject.

In one embodiment, the heterozygous rs12459419T genotype has been detected in a sample obtained from the subject. In one embodiment, the method of treating cancer comprises detecting a heterozygous rs12459419T genotype in a sample obtained from a subject.

In one embodiment, the sample obtained from the subject is a blood sample or a buccal swab.

In one embodiment, the cancer is characterized by a RAS mutation. In one embodiment, the cancer is characterized by a TP53 mutation. In one embodiment, the cancer is characterized by an IDH mutation. In one embodiment, the cancer is characterized by a FLT3 mutation.

In one embodiment, the cancer is chemotherapy sensitive.

In one embodiment, the cancer is chemoresistant.

In one embodiment, at least 20% of the cancer-derived blasts are CD33 positive, as measured by flow cytometry.

In one embodiment, the anti-CD33 immunoconjugate (eg, IMGN779) is administered at a dose (eg, 0.39 mg/kg or 0.54 mg) that maintains exposure to the anti-CD33 immunoconjugate, eg, over at least 7 days after infusion. /Kg; for example, once a week or once every two weeks).

In one embodiment, the administration results in saturation of free CD33.

In one embodiment, the administration results in a reduction in peripheral hemoblasts, eg, within 3-8 days of the first administration of the anti-CD33 immunoconjugate (eg, IMGN779) or after the second administration.

In one embodiment, the administration results in a reduction in myeloblasts (eg, at least 45%, at least 48%, at least 59%, or at least 96% reduction from baseline).

In one embodiment, the subject is a human.

In one embodiment, the method of treating AML in humans comprises:
The following formula:

About 0.7, 0.75, 0.8, 0.85, 0.9, 0.91, 0.92, 0.93, 0 of the anti-CD33 immunoconjugate or pharmaceutically acceptable salt thereof of .94, 0.95, 0.96, 0.97, 0.98, 0.99, or 1.0 mg/kg administered to the subject about once every two weeks, wherein: Double line between C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a comprises a light chain comprising the sequence set forth in SEQ and SEQ ID NO: 12 as set forth in SEQ ID NO: 11, anti-CD33 antibody or antigen-binding It is a fragment.

In one embodiment, the method of treating AML in humans comprises:
The following formula:

About 1.1, 1.11, 1.12, 1.13, 1.14, 1.15, 1.16, 1.17, 1 of the anti-CD33 immunoconjugate or a pharmaceutically acceptable salt thereof of .18, 1.19, 1.2, 1.21, 1.22, 1.23, 1.24, or 1.25 mg/kg to the subject about once every two weeks, Middle: Double line between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a comprises a light chain comprising the sequence set forth in SEQ and SEQ ID NO: 12 as set forth in SEQ ID NO: 11, anti-CD33 antibody or antigen-binding It is a fragment.

In one embodiment, the method of treating AML in humans comprises:
The following formula:

1.50, 1.51, 1.52, 1.53, 1.54, 1.55, 1.56, 1.57, 1, of an anti-CD33 immunoconjugate of 0.58, 1.59, 1.6, or 2.0 mg/kg administered to the subject about once every two weeks, wherein: the doublet between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a comprises a light chain comprising the sequence set forth in SEQ and SEQ ID NO: 12 as set forth in SEQ ID NO: 11, anti-CD33 antibody or antigen-binding It is a fragment.

In one embodiment, the method of treating AML in humans comprises:
The following formula:

About 0.45, 0.46, 0.47, 0.48, 0.49, 0.5, 0.51, 0.52, 0 of the anti-CD33 immunoconjugate or pharmaceutically acceptable salt thereof of .53, 0.54, 0.55, 0.56, 0.57, 0.58, 0.59, 0.6, 0.61, 0.62, 0.63, 0.64, 0.65 , 0.66, 0.67, 0.68, 0.69, or 0.7 mg/kg administered to the subject once weekly, wherein: the doublet between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a comprises a light chain comprising the sequence set forth in SEQ and SEQ ID NO: 12 as set forth in SEQ ID NO: 11, anti-CD33 antibody or antigen-binding It is a fragment.

In one embodiment, r is an integer from 2-9, 3-8, 4-7, or 5-6.

In one embodiment, the indolinobenzodiazepine dimer (eg, DGN462) is linked to the anti-CD33 antibody or antigen binding fragment thereof via the lysine residue of the antibody or antigen binding fragment thereof. In one embodiment, 1-10, 2-9, 3-8, 4-7, or 5-6 indolinobenzodiazepine dimer molecules (eg, DGN462) are lysine residues of the antibody or antigen-binding fragment thereof. Via anti-CD33 antibody or an antigen-binding fragment thereof.

In one example (I1), the method of treating cancer in a subject is
The following formula:

Comprising administering to the subject about 0.54 to about 0.8 mg/kg of an anti-CD33 immunoconjugate or a pharmaceutically acceptable salt thereof, wherein: the doublet between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is a There integer from 1 to 10; a comprises a light chain comprising the sequence set forth in the heavy chain and SEQ ID NO: 12 comprises the sequence set forth in SEQ ID NO: 11, anti-CD33 antibody Alternatively, it is an antigen-binding fragment thereof.

In one example of I1 (I2), about 0.54 to about 0.70 mg/kg of the immunoconjugate is administered. In one example of I1 (I3), about 0.54 or about 0.70 mg/kg of the immunoconjugate is administered.

In one example (I4) of any one of I1 to I3, the immunoconjugate is administered about once every two weeks.

In one example (I5), the method of treating cancer in a subject is
The following formula:

Comprising administering to the subject about 0.30 to about 0.54 mg/kg of an anti-CD33 immunoconjugate or a pharmaceutically acceptable salt thereof, wherein: a doublet between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is a There integer from 1 to 10; a comprises a light chain comprising the sequence set forth in the heavy chain and SEQ ID NO: 12 comprises the sequence set forth in SEQ ID NO: 11, anti-CD33 antibody Alternatively, it is an antigen-binding fragment thereof.

In one example of I5 (I6), about 0.39 to about 0.54 mg/kg of the immunoconjugate is administered. In one example of I5 (I7), about 0.39 or about 0.54 mg/kg of the immunoconjugate is administered. In one example (I8) of any one of I5 to I7, the immunoconjugate is administered about once a week.

In one example (I9) of any one of I1 to I8, the immunoconjugate is administered in a 28-day cycle. In one example of any one of I1-I9 (I10), the immunoconjugate is administered by intravenous infusion. In one example of any one of I1 to I10 (I11), the variable heavy chain comprises the sequence set forth in SEQ ID NO:9, and the variable light chain comprises the sequence set forth in SEQ ID NO:10. In one example of I11 (I12), the antibody or antigen-binding fragment thereof comprises a heavy chain comprising the sequence set forth in SEQ ID NO:11 and a light chain comprising the sequence set forth in SEQ ID NO:12.

In one example (I13) of any one of I1 to I12, the immunoconjugate comprises
Formula (II):

Or an pharmaceutically acceptable salt thereof, wherein A is an anti-CD33 antibody or an antigen-binding fragment thereof, and r is an integer of 1 to 10.

In one example (I14) of any one of I1 to I12, the immunoconjugate comprises
Formula (III):

Or an pharmaceutically acceptable salt thereof, wherein A is an anti-CD33 antibody or an antigen-binding fragment thereof, and r is an integer of 1 to 10.

In one example (I15) of any one of I1 to I14, the pharmaceutically acceptable salt is a sodium or potassium salt. In one example (I16) of any one of I1 to I15, r is an integer of 2 to 9, 3 to 8, 4 to 7, or 5 to 6. In one example of any one of I1 to I15 (I17), the immunoconjugate is included in a composition comprising at least two of the immunoconjugates, and the average number of cytotoxins per antibody or antigen-binding fragment thereof. Is 2 to 8, 3 to 7, 3 to 5, or 2.5 to 3.5. In one example of any one of I1 to I17 (I18), the immunoconjugate is IMGN779.

In one example of any one of I1 to I18 (I19), the cancer is selected from the group consisting of leukemia, lymphoma, and myeloma. In one example of I19 (I20), the cancer is acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute lymphoblastic leukemia (ALL), B cell lineage of acute lymphoblastic leukemia (B). ALL), T cell acute lymphoblastic leukemia (T ALL), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), myelodysplastic syndrome (MDS), basic plasmacytoid DC tumor (BPDCN). ) Selected from the group consisting of leukemia, non-Hodgkin's lymphoma (NHL), mantle cell lymphoma, and Hodgkin's leukemia (HL). In one example of I20 (I21), the cancer is acute myelogenous leukemia (AML). In one example of I21 (I22), AML is refractory or relapsed acute myeloid leukemia. In one example of I21 or I22 (I23), AML expresses P-glycoprotein overexpression, EVI1 overexpression, p53 mutation, DNMT3A mutation, FLT3 internal tandem duplication, complex karyotype, BRCA1, BRCA2, or PALB2. It is characterized by a decrease, or a mutation in BRCA1, BRCA2, or PALB2.

In one example of any one of I1-I23 (I24), the homozygous rs12459419C genotype has been detected in a sample obtained from the subject. In one example of I24 (I25), the method further comprises detecting the homozygous rs12459419C genotype in a sample obtained from the subject. In one example of any one of I1-I23 (I26), the heterozygous rs12459419C genotype has been detected in a sample obtained from the subject. In one example of I26 (I27), the method further comprises detecting the heterozygous rs12459419C genotype in a sample obtained from the subject. In one example of any one of I1-I23 (I28), the homozygous rs12459419T genotype has been detected in a sample obtained from the subject. In one example of I28 (I29), the method further comprises detecting the homozygous rs12459419T genotype in a sample obtained from the subject. In one example of any one of I1-I23 (I30), the heterozygous rs12459419T genotype has been detected in a sample obtained from the subject. In one example of I30 (I31), the method further comprises detecting the heterozygous rs12459419T genotype in a sample obtained from the subject. In one example of any one of I24-I31 (I32), the sample obtained from the patient is a blood sample or a buccal swab.

In one example of any one of I1-I32 (I33), the cancer is chemosensitive. In one example of any one of I1-I32 (I34), the cancer is chemo-resistant. In one example of any one of I1-I34 (I35), at least 20% of the cancer-derived blasts are CD33-positive, as measured by flow cytometry.

In one example of any one of I1-I35 (I36), the administration results in saturation of free CD33.

In one example of any one of I1-I36 (I37), the subject is a human.

In one example (I38), a method of treating AML in humans comprises:
The following formula:

Comprising administering to the subject about 0.7 mg/kg of the anti-CD33 immunoconjugate or pharmaceutically acceptable salt thereof about once every two weeks, wherein the doublet between N and C.

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is a There integer from 1 to 10; a comprises a light chain comprising the sequence set forth in the heavy chain and SEQ ID NO: 12 comprises the sequence set forth in SEQ ID NO: 11, anti-CD33 antibody Alternatively, it is an antigen-binding fragment thereof.

In one example (I39), a method of treating AML in humans comprises:
The following formula:

Comprising administering to the subject about 0.54 mg/kg of an anti-CD33 immunoconjugate or a pharmaceutically acceptable salt thereof once a week, wherein the doublet between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is a There integer from 1 to 10; a comprises a light chain comprising the sequence set forth in the heavy chain and SEQ ID NO: 12 comprises the sequence set forth in SEQ ID NO: 11, anti-CD33 antibody Alternatively, it is an antigen-binding fragment thereof.

In one example (I40) of I38 or I39, r is an integer of 2 to 9, 3 to 8, 4 to 7, or 5 to 6.

In one example (I41), the method of treating cancer in a subject is
The following formula:

Comprising administering to the subject about 0.7 to about 1 mg/kg of an anti-CD33 immunoconjugate of or a pharmaceutically acceptable salt thereof, wherein the doublet between N and C:

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a is variable heavy chain comprising the CDRl, CDR2, and CDR3 sequences set forth in SEQ ID NO: 1-3, and SEQ ID NO: 4-6 An anti-CD33 antibody or an antigen-binding fragment thereof, which comprises a variable light chain comprising the CDR1, CDR2, and CDR3 sequences described, respectively.

In one example of I41 (I42), about 0.7 to about 0.9 mg/kg of the immunoconjugate is administered. In one example of I41 (I43), about 0.7 mg/kg of the immunoconjugate is administered. In one example of I41 (I44), about 0.75 mg/kg of the immunoconjugate is administered. In one example of I41 (I45), about 0.8 mg/kg of the immunoconjugate is administered. In one example of I41 (I46), about 0.85 mg/kg of the immunoconjugate is administered. In one example of I41 (I47), about 0.9 mg/kg of the immunoconjugate is administered. In one example of I41 (I48), about 0.95 mg/kg of the immunoconjugate is administered. In one example of I41 (I49), about 1 mg/kg of the immunoconjugate is administered. In one example of any one of I41 to I49 (I50), the immunoconjugate is administered about once every two weeks.

In one example (I51), the method of treating cancer in a subject is
The following formula:

Comprising administering to the subject about 0.39 to about 0.7 mg/kg of an anti-CD33 immunoconjugate or a pharmaceutically acceptable salt thereof, wherein: a doublet between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a is variable heavy chain comprising the CDRl, CDR2, and CDR3 sequences set forth in SEQ ID NO: 1-3, and SEQ ID NO: 4-6 An anti-CD33 antibody or an antigen-binding fragment thereof, which comprises a variable light chain comprising the CDR1, CDR2, and CDR3 sequences described, respectively.

In one example of I51 (I52), about 0.39 mg/kg of the immunoconjugate is administered. In one example of I51 (I53), about 0.45 mg/kg of immunoconjugate is administered. In one example of I51 (I54), about 0.5 mg/kg of immunoconjugate is administered. In one example of I51 (I55), about 0.54 mg/kg of immunoconjugate is administered. In one example of I51 (I56), about 0.55 mg/kg of immunoconjugate is administered. In one example of I51 (I57), about 0.6 mg/kg of the immunoconjugate is administered. In one example of I51 (I58), about 0.65 mg/kg of the immunoconjugate is administered. In one example of I51 (I59), about 0.7 mg/kg of the immunoconjugate is administered. In one example of any one of I51 to I59 (I60), the immunoconjugate is administered about once a week.

In one example of any one of I41 to I60 (I61), the immunoconjugate is administered in a 28-day cycle. In one example of any one of I41-I61 (I62), the immunoconjugate is administered by intravenous infusion. In an example of any one of I41 to I62 (I63), the variable heavy chain comprises the sequence set forth in SEQ ID NO:9 and the variable light chain comprises the sequence set forth in SEQ ID NO:10. In one example of I63 (I64), the antibody or antigen-binding fragment thereof comprises a heavy chain comprising the sequence set forth in SEQ ID NO:11 and a light chain comprising the sequence set forth in SEQ ID NO:12. In one example (I65) of any one of I41 to I64, the immunoconjugate is
Formula (II):

Or an pharmaceutically acceptable salt thereof, wherein A is an anti-CD33 antibody or an antigen-binding fragment thereof, and r is an integer of 1 to 10.

In one example (I66) of any one of I41 to I64, the immunoconjugate comprises
Formula (III):

Or an pharmaceutically acceptable salt thereof, wherein A is an anti-CD33 antibody or an antigen-binding fragment thereof, and r is an integer of 1 to 10.

In one example (I67) of any one of I41 to I66, the pharmaceutically acceptable salt is a sodium or potassium salt. In one example (I68) of any one of I41 to I67, r is an integer of 2 to 9, 3 to 8, 4 to 7, or 5 to 6. In one example of any one of I41-I67 (I69), the immunoconjugate is included in a composition comprising at least two of the immunoconjugates, and the average number of cytotoxins per antibody or antigen-binding fragment thereof is included. Is 2 to 8, 3 to 7, 3 to 5, or 2.5 to 3.5. In one example of any one of I41 to I69 (I70), the immunoconjugate is IMGN779. In one example of any one of I41 to I70 (I71), the cancer is selected from the group consisting of leukemia, lymphoma, and myeloma. In one example of I71 (I72), the cancer is acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute lymphoblastic leukemia (ALL), B cell lineage of acute lymphoblastic leukemia (B). ALL), T cell acute lymphoblastic leukemia (T ALL), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), myelodysplastic syndrome (MDS), basic plasmacytoid DC tumor (BPDCN). ) Selected from the group consisting of leukemia, non-Hodgkin's lymphoma (NHL), mantle cell lymphoma, and Hodgkin's leukemia (HL). In one example of I72 (I73), the cancer is acute myelogenous leukemia (AML). In one example of I73 (I74), AML is refractory or relapsed acute myelogenous leukemia. In one example of I73 (I75), the AML is a newly diagnosed AML. In one example of any one of I71 to I75 (I76), AML has P-glycoprotein overexpression, EVI1 overexpression, p53 mutation, DNMT3A mutation, FLT3 internal tandem duplication, complex karyotype, BRCA1, BRCA2, Alternatively, it is characterized by reduced expression of PALB2, or a mutation of BRCA1, BRCA2, or PALB2. In one example of I72 (I77), the MDS is a high risk MDS.

In one example of any one of I41 to I77 (I78), the homozygous rs12459419C genotype has been detected in a sample obtained from the subject. In one example of I78 (I79), the method further comprises detecting the homozygous rs12459419C genotype in a sample obtained from the subject. In one example of any one of I41 to I77 (I80), the heterozygous rs12459419C genotype has been detected in a sample obtained from the subject. In one example of I80 (I81), the method further comprises detecting the heterozygous rs12459419C genotype in a sample obtained from the subject. In one example of any one of I41 to I77 (I82), the homozygous rs12459419T genotype has been detected in a sample obtained from the subject. In one example of I82 (I83), the method further comprises detecting the homozygous rs12459419T genotype in a sample obtained from the subject. In one example (I84) of any one of I41 to I77, the heterozygous rs12459419T genotype has been detected in a sample obtained from the subject. In one example of I84 (I85), the method further comprises detecting the heterozygous rs12459419T genotype in a sample obtained from the subject. In one example of any one of I78-I85 (I86), the sample obtained from the patient is a blood sample or a buccal swab.

In one example of any one of I41-I86 (I87), the cancer is chemosensitive. In one example of any one of I41-I86 (I88), the cancer is chemoresistant. In one example of any one of I41-I88 (I89), at least 20% of the cancer-derived blasts are CD33-positive, as measured by flow cytometry. In one example of any one of I41-I89 (I90), the administration results in saturation of free CD33.

In one example of any one of I41-I90 (I91), the administration results in a reduction in peripheral hemoblasts. In one example of any one of I41-I91 (I92), the administration results in a reduction in myeloblasts. In one example of any one of I41 to I90 (I93), the subject is a human.

In one example (I94), the method of treating cancer in a subject is
The following formula:

Comprising administering to the subject about 0.7 to about 1.25 mg/kg of the anti-CD33 immunoconjugate or a pharmaceutically acceptable salt thereof, wherein the doublet between N and C:

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a is variable heavy chain comprising the CDRl, CDR2, and CDR3 sequences set forth in SEQ ID NO: 1-3, and SEQ ID NO: 4-6 An anti-CD33 antibody or an antigen-binding fragment thereof, which comprises a variable light chain comprising the CDR1, CDR2, and CDR3 sequences described, respectively.

In one example of I94 (I95), about 0.7 to about 1 mg/kg of the immunoconjugate is administered. In one example of I94 (I96), about 0.7 to about 0.9 mg/kg of the immunoconjugate is administered. In one example of I94 (I97), about 0.7 mg/kg of the immunoconjugate is administered. In one example of I94 (I98), about 0.75 mg/kg of immunoconjugate is administered. In one example of I94 (I99), about 0.8 mg/kg of immunoconjugate is administered. In one example of I94 (I100), about 0.85 mg/kg of the immunoconjugate is administered. In one example of I94 (I101), about 0.9 mg/kg of immunoconjugate is administered. In one example of I94 (I102), about 0.95 mg/kg of immunoconjugate is administered. In one example of I94 (I103), about 1 mg/kg of immunoconjugate is administered.

In any one example of I94-I103 (I104), the immunoconjugate is administered about once a week.

In one example of I94 (I105), about 1 to about 1.25 mg/kg of the immunoconjugate is administered. In one example of I94 (I106), about 1.1 mg/kg of the immunoconjugate is administered. In one example of I94 (I107), about 1.15 mg/kg of the immunoconjugate is administered. In one example of I94 (I108), about 1.2 mg/kg of the immunoconjugate is administered. In one example of I94 (I109), about 1.25 mg/kg of immunoconjugate is administered. In one example of any one of I94 and I105 to I109 (I110), the immunoconjugate is administered about once every two weeks. In one example of any one of I94 and I105 to I109 (I111), the immunoconjugate is administered about once a week.

In any one of I94 to I111 (I112), the immunoconjugate is administered in a 28-day cycle. In one example of any one of I94-I112 (I113), the immunoconjugate is administered by intravenous infusion. In one example of any one of I94 to I113 (I114), the variable heavy chain comprises the sequence set forth in SEQ ID NO:9 and the variable light chain comprises the sequence set forth in SEQ ID NO:10. In one example of I114 (I115), the antibody or antigen-binding fragment thereof comprises a heavy chain comprising the sequence set forth in SEQ ID NO:11 and a light chain comprising the sequence set forth in SEQ ID NO:12.

In one example (I116) of any one of I94 to I115, the immunoconjugate comprises
Formula (II):

Or an pharmaceutically acceptable salt thereof, wherein A is an anti-CD33 antibody or an antigen-binding fragment thereof, and r is an integer of 1 to 10.

In one example (I117) of any one of I94 to I115, the immunoconjugate is:
Formula (III):

Or an pharmaceutically acceptable salt thereof, wherein A is an anti-CD33 antibody or an antigen-binding fragment thereof, and r is an integer of 1 to 10.

In one example of any one of I94 to I17 (I118), the pharmaceutically acceptable salt is a sodium or potassium salt. In one example (I119) of any one of I94 to I118, r is an integer of 2 to 9, 3 to 8, 4 to 7, or 5 to 6. In one example of any one of I94 to I118 (I120), the immunoconjugate is included in a composition comprising at least two of the immunoconjugates and the average number of cytotoxins per antibody or antigen-binding fragment thereof. Is 2 to 8, 3 to 7, 3 to 5, or 2.5 to 3.5. In one example of any one of I94 to I120 (I121), the immunoconjugate is IMGN779.

In one example of any one of I94 to I121 (I122), the cancer is selected from the group consisting of leukemia, lymphoma, and myeloma. In one example of I122 (I123), the cancer is acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute lymphoblastic leukemia (ALL), B cell lineage of acute lymphoblastic leukemia (B). ALL), T cell acute lymphoblastic leukemia (T ALL), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), myelodysplastic syndrome (MDS), basic plasmacytoid DC tumor (BPDCN). ) Selected from the group consisting of leukemia, non-Hodgkin's lymphoma (NHL), mantle cell lymphoma, and Hodgkin's leukemia (HL). In one example of I123 (I124), the cancer is acute myelogenous leukemia (AML). In one example of I124 (I125), AML is refractory or relapsed acute myelogenous leukemia. In one example of I124 (I126), the AML is a newly diagnosed AML. In any one example of I124-I126 (I127), AML is P-glycoprotein overexpression, EVI1 overexpression, p53 mutation, DNMT3A mutation, FLT3 internal tandem duplication, complex karyotype, BRCA1, BRCA2, Alternatively, it is characterized by reduced expression of PALB2, or a mutation of BRCA1, BRCA2, or PALB2. In one example of I123 (I128), the MDS is a high risk MDS.

In one example of any one of I94 to I128 (I129), the homozygous rs12459419C genotype has been detected in a sample obtained from the subject. In one example of I129 (I130), the method further comprises detecting the homozygous rs12459419C genotype in a sample obtained from the subject. In one example of any one of I94 to I128 (I131), the heterozygous rs12459419C genotype has been detected in a sample obtained from the subject. In one example of I131 (I132), the method further comprises detecting the heterozygous rs12459419C genotype in a sample obtained from the subject. In one example of any one of I94-I128 (I133), the homozygous rs12459419T genotype has been detected in a sample obtained from the subject. In one example of I133 (I134), the method further comprises detecting the homozygous rs12459419T genotype in a sample obtained from the subject. In one example of any of I94-I128 (I135), the heterozygous rs12459419T genotype has been detected in a sample obtained from the subject. In one example of I135 (I136), the method further comprises detecting the heterozygous rs12459419T genotype in a sample obtained from the subject. In the method of any one of I129-I136 (I137), any of claims 129-136, the sample obtained from the patient is a blood sample or a buccal swab.

In one example (I138) of any one of I94 to I137, the cancer is chemosensitive. In one example (I139) of any one of I94 to I137, the cancer is chemoresistant. In one example of any one of I94-I139 (I140), at least 20% of the cancer-derived blasts are CD33-positive, as measured by flow cytometry.

In one example of any one of I94-I140 (I141), the administration results in saturation of free CD33. In one example of any one of I94-I141 (I142), the administration results in a reduction in peripheral hemoblasts. In one example of any one of I94-I142 (I143), the administration results in a reduction in myeloblasts. In one example of any one of I94 to I143 (I144), the subject is a human.

In one example (I145), the method of treating cancer is
The following formula:

Comprising administering to the subject about 1 to about 2 mg/kg of an anti-CD33 immunoconjugate of or a pharmaceutically acceptable salt thereof, wherein the doublet between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is be a -SO ₃ H; r is an integer from 1 to 10; a is variable heavy chain comprising the CDRl, CDR2, and CDR3 sequences set forth in SEQ ID NO: 1-3, and SEQ ID NO: 4-6 An anti-CD33 antibody or an antigen-binding fragment thereof, which comprises a variable light chain comprising the CDR1, CDR2, and CDR3 sequences described, respectively.

In one example of I145 (I146), about 1.2 to about 2 mg/kg of immunoconjugate is administered. In one example of I145 (I147), about 1.5 to about 1.6 mg/kg mg/kg of the immunoconjugate is administered. In one example of I145 (I148), about 1.5 mg/kg or about 1.56 mg/kg of immunoconjugate is administered. In one example of any one of I145-I148 (I149), the immunoconjugate is administered about once every two weeks. In one example of any one of I145-I148 (I150), the immunoconjugate is administered about once a week.

In one example of any one of I145 to I150 (I151), the immunoconjugate is administered in a 28-day cycle. In one example of any one of I145-I151 (I152), the immunoconjugate is administered by intravenous infusion. In one example of any one of I145 to I152 (I153), the variable heavy chain comprises the sequence set forth in SEQ ID NO:9 and the variable light chain comprises the sequence set forth in SEQ ID NO:10. In one example of I153 (I154), the antibody or antigen-binding fragment thereof comprises a heavy chain comprising the sequence set forth in SEQ ID NO:11 and a light chain comprising the sequence set forth in SEQ ID NO:12.

In one example of any one of I145 to I154 (I155), the immunoconjugate comprises
Formula (II):

Or an pharmaceutically acceptable salt thereof, wherein A is an anti-CD33 antibody or an antigen-binding fragment thereof, and r is an integer of 1 to 10.

In one example (I156) of any one of I145 to I154, the immunoconjugate comprises
Formula (III):

Or an pharmaceutically acceptable salt thereof, wherein A is an anti-CD33 antibody or an antigen-binding fragment thereof, and r is an integer of 1 to 10.

In one example of any one of I145-I156 (I157), the pharmaceutically acceptable salt is a sodium or potassium salt. In one example (I158) of any one of I145 to I157, r is an integer of 2 to 9, 3 to 8, 4 to 7, or 5 to 6. In one example of any one of I145-I157 (I159), the immunoconjugate is included in a composition comprising at least two of the immunoconjugates, and the average number of cytotoxins per antibody or antigen-binding fragment thereof is included. Is 2 to 8, 3 to 7, 3 to 5, or 2.5 to 3.5. In one example of any one of I145 to I159 (I160), the immunoconjugate is IMGN779.

In one example of any one of I145 to I160 (I161), the cancer is selected from the group consisting of leukemia, lymphoma, and myeloma. In one example of I161 (I162), the cancer is acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute lymphoblastic leukemia (ALL), B cell lineage of acute lymphoblastic leukemia (B). ALL), T cell acute lymphoblastic leukemia (T ALL), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), myelodysplastic syndrome (MDS), basic plasmacytoid DC tumor (BPDCN). ) Selected from the group consisting of leukemia, non-Hodgkin's lymphoma (NHL), mantle cell lymphoma, and Hodgkin's leukemia (HL). In one example of I162 (I163), the cancer is acute myelogenous leukemia (AML). In one example of I163 (I164), AML is refractory or relapsed acute myelogenous leukemia. In an example of I163 (I165), the AML is a newly diagnosed AML. In any one example of I163-I165 (I166), AML is P-glycoprotein overexpression, EVI1 overexpression, p53 mutation, DNMT3A mutation, FLT3 internal tandem duplication, complex karyotype, BRCA1, BRCA2, Alternatively, it is characterized by reduced expression of PALB2, or a mutation of BRCA1, BRCA2, or PALB2. In one example of I162 (I167), the MDS is a high risk MDS.

In one example of any one of I145-I167 (I168), the homozygous rs12459419C genotype has been detected in a sample obtained from the subject. In one example of I168 (I169), the method further comprises detecting the homozygous rs12459419C genotype in a sample obtained from the subject. In one example of any one of I145-I167 (I170), the heterozygous rs12459419C genotype has been detected in a sample obtained from the subject. In one example of I170 (I171), the method further comprises detecting the heterozygous rs12459419C genotype in a sample obtained from the subject. In one example of any one of I145-I167 (I172), the homozygous rs12459419T genotype has been detected in a sample obtained from the subject. In one example of I172 (I173), the method further comprises detecting the homozygous rs12459419T genotype in a sample obtained from the subject. In one example of any one of I145-I167 (I174), the heterozygous rs12459419T genotype has been detected in a sample obtained from the subject. In one example of I174 (I175), the method further comprises detecting the heterozygous rs12459419T genotype in a sample obtained from the subject. In one example of any one of I168-I175 (I176), the sample obtained from the patient is a blood sample or a buccal swab.

In one example of any one of I145-I176 (I177), the cancer is chemosensitive. In one example of any one of I145-I176 (I178), the cancer is chemoresistant. In one example of any one of I145-I178 (I179), at least 20% of the cancer-derived blasts are CD33-positive, as measured by flow cytometry.

In one example of any one of I145-I179 (I180), the administration results in saturation of free CD33. In one example of any one of I145-I180 (I181), the administration results in a reduction in peripheral hemoblasts. In one example of any one of I145-I181 (I182), the administration results in a reduction in myeloblasts. In one example of any one of I145 to I182 (I183), the subject is a human.

In one example of any one of I1-I183 (I184), the cancer is characterized by a RAS mutation. In one example of any one of I1-I184 (I185), the cancer is characterized by a TP53 mutation. In one example of any one of I1-I185 (I186), the cancer is characterized by an IDH mutation. In one example of any one of I1-I186 (I187), the cancer is characterized by an FLT3 mutation.

FIG. 1A shows the in vitro potency of IMGN779 in the EOL-1 cell line. FIG. 1B shows the in vitro potency of IMGN779 in the MV4-11 cell line. In vitro potency of DGN462 and DGN484 in three AML cell lines. 6 shows the apoptotic activity of DGN462 and DGN484 in three AML cell lines. Figure 7 shows indirect binding of CD33 antibody drug conjugate to MOLM-13 cells. 8 shows the in vitro potency of IMGN779 and Z4681A-sSPDB-DGN484 in MOLM-13 cell line. 8 shows the in vitro potency of IMGN779 and Z4681A-sSPDB-DGN484 in the MV4-11 cell line. 8 shows the in vitro potency of IMGN779 and Z4681A-sSPDB-DGN484 in the EOL-1 cell line. 8 shows the in vitro potency of IMGN779 and Z4681A-sSPDB-DGN484 in HNT-34. FIG. 6 shows the results of a single intravenous dose tolerability study of IMGN779 in female CD-1 mice. Figure 3 shows the results of an intravenous single dose tolerability study of IMGN779 and Z4681A-sSPDB-DGN484 conjugates in female CD-1 mice. 5 shows the results of in vivo efficacy studies of a single dose of IMGN779 in the EOL-1 subcutaneous model. 5 shows the results of in vivo efficacy studies of a single dose of IMGN779 in the HL60/QC subcutaneous model. Figure 7 shows the results of single and multiple dose in vivo efficacy studies of IMGN779 in the HL60/QC subcutaneous model. Figure 7 shows the results of a single dose in vivo efficacy study of IMGN779 and Z4681A-sSPDB-DGN484 in the HL60/QC subcutaneous model. Figure 7 shows the results of a single versus multiple divided dose in vivo efficacy study in IMGN79 in the MV4-11 subcutaneous model. Figure 7 shows the results of in vivo efficacy studies of a single dose of IMGN779 in the MV4-11 disseminated model. Figure 7 shows the results of in vivo efficacy studies of multiple doses of IMGN779 in the MV4-11 disseminated model. Figure 7 shows the results of in vivo efficacy studies of single dose IMGN779 and Z6481A-sSPDB-DGN484 in the MV4-11 disseminated model. Figure 3 shows the results of in vivo efficacy studies of multiple doses of IMGN779 in the Molm-13 disseminated model. 3 shows demographics of 23 patients treated with IMGN779 (0.02 mg/kg to 0.54 mg/kg). Figure 7 shows adverse events occurring during treatment that occurred at a frequency of at least 10% in 23 patients treated with IMGN779 (0.02 mg/kg to 0.54 mg/kg). Shows adverse events occurring in at least 2 of 23 patients of grade 3 or higher and treated with IMGN779 (0.02 mg/kg to 0.54 mg/kg). Shows the change (%) from baseline in absolute peripheral sprouting in patients in cohorts 6 and 7. Shows the maximum change from baseline in myeloblasts in patients in cohorts 6 and 7. A shows IMGN779 concentration over time at various doses of IMGN779. B shows C _max values calculated for cycle 1 (first dosing cycle per patient) at various doses of IMGN779. C shows the calculated AUC for cycle 1 (first dosing cycle per patient) at various doses of IMGN779. A shows IMGN779 concentration over time at various doses of IMGN779. B shows C _max values calculated for cycle 1 (first dosing cycle per patient) at various doses of IMGN779. C shows the calculated AUC for cycle 1 (first dosing cycle per patient) at various doses of IMGN779. IMGN779 (left panel) concentration and CD33 saturation (right panel) in patients in cohorts 4-7. Shows IMGN779 exposure (left panel) and CD33 saturation (right panel) in patients treated with 0.04-0.91 mg/kg IMGN779. CD33 saturation (left) and end of infusion concentration (right) in patients treated weekly (QW) or biweekly (Q2W) with 0.39 mg/kg IMGN779. Figure 6 shows the rate of myeloblast changes in patients treated with IMGN779. Figure 7 shows adverse events (TEAEs) occurring during Grade 3+ treatment that occurred in 2 or more patients after IMGN779 administration. 27A and 27B show the concentration of IMGN779 in the blood of patients after receiving various doses. Figure 27A shows the concentration of ADC (IMGN779) throughout cycle 1 (2 weeks) at various doses, and Figure 27B relates to administration at 0.39 mg/kg Q2W and 0.39 mg/kg QW. 8 shows IMGN779 concentrations at the end of infusion in various patients. 28A and 28B show the pharmacodynamics associated with CD33 saturation at various doses. FIG. 28A shows the availability of CD33 receptors in blasts from patients treated with various concentrations of IMGN779 on the Q2W schedule, and FIG. 28B shows blasts from patients treated with various concentrations of IMGN779 on the QW schedule. 2 shows the availability of the CD33 receptor in the medium. 29A and 29B show a comparison of Q2W and QW dosing schedules at a dose of 0.54 mg/kg. FIG. 29A shows the availability of CD33 receptors in blasts from patients treated with 0.54 mg/kg IMGN779 on the Q2W schedule, and FIG. 29B shows patients treated with 0.54 mg/kg IMGN779 on the QW schedule. Shows the availability of the CD33 receptor in the derived blasts. 30A and 30B show CD33 expression in leukemic cells obtained from patients treated with IMGN779. FIG. 30A shows CD33 levels plotted against binding sites per cell in patients from cohort 6+, and FIG. 30B shows percent CD33+ leukemic cells in local bone marrow. Asterisk indicates formal or informal bone marrow response (more than 25% reduction in myeloblast+peripheral blood (PB) clearance). The average CD33 antigen per cell of patients with three different SNP allele combinations is shown. Asterisk indicates formal or informal bone marrow response (more than 25% reduction in myeloblast+PB clearance). Figure 6 shows the best response obtained in patients treated with various dosing schedules of IMGN779. Shown are the elimination rate constant estimates obtained from elimination half-life data for plasma ADC (IMGN779) at various doses. It shows that the linear pharmacokinetic (PK) model does not fit the PK data of IMGN779. Figures 35A and 35B show IMGN779 efflux rate constants related to the number of binding sites on target cells (Figure 35A) and total target concentration (Figure 35B). It shows that the target-mediated pharmacokinetic model does not fit the PK data of IMGN779. 37A and 37B show scatter plots (FIG. 37A) and box plots (FIG. 37B) of measured and estimated subjects in patients treated with IMGN779. We show that the binding model fits better with the IMGN779 PK data.

I. Definitions To facilitate an understanding of the present invention, a number of terms and phrases are defined below.

The term "CD33", as used herein, unless otherwise indicated, refers to any naturally occurring CD33 polypeptide. The term "CD33" includes "full length" unprocessed CD33 polypeptide as well as any form of CD33 resulting from intracellular processing (eg, removal of the signal peptide). The term also includes naturally occurring variants of CD33, such as those encoded by splice variants and allelic variants. The CD33 polypeptides described herein can be isolated from a variety of sources, such as human tissue type or another source, or can be prepared by recombinant or synthetic methods. Unless otherwise specified, "CD33" can be used to refer to a nucleic acid that encodes a CD33 polypeptide. Human CD33 sequences are known and include, for example, the publicly available sequences (including isoforms) under NCBI Accession No. CAD36509. As used herein, the term "human CD33" refers to CD33 comprising the sequence of amino acids 18-364 of SEQ ID NO:14:

The term “antibody” recognizes a target, such as a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid or combination of the foregoing, via at least one antigen recognition site within the variable region of an immunoglobulin molecule. , An immunoglobulin molecule that specifically binds to it. As used herein, the term "antibody" refers to an intact polyclonal antibody, an intact monoclonal antibody, a chimeric antibody, a humanized antibody, a human antibody, a fusion protein containing an antibody, as long as the antibody exhibits the desired biological activity, Also included are any other modified immunoglobulin molecules. Antibodies are classified into five major classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, or their subclasses (isotypes) based on the identity of the heavy chain constant domains called α, δ, ε, γ and μ, respectively. ) (Eg, IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2). Different classes of immunoglobulins have different well-known subunit and three-dimensional structures. Antibodies can be naked or conjugated to other molecules such as toxins, radioisotopes and the like.

The term "antibody fragment" refers to a portion of an intact antibody. “Antigen-binding fragment” refers to the portion of an intact antibody that binds an antigen. The antigen binding fragment may comprise the antigen-determining variable region of an intact antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2 and Fv fragments, linear antibodies, and single chain antibodies. Antibody fragments can be naked or conjugated to other molecules such as toxins, radioisotopes and the like.

The term "anti-CD33 antibody" or "antibody that binds to CD33" refers to an antibody capable of binding to CD33 with sufficient affinity, such that the antibody is a diagnostic and/or therapeutic agent that targets CD33. Is useful as The extent to which an anti-CD33 antibody binds to an unrelated non-CD33 protein can be less than about 10% of the antibody's binding to CD33, as measured by, for example, radioimmunoassay (RIA).

A "monoclonal" antibody or antigen-binding fragment thereof refers to a homogeneous population of antibodies or antigen-binding fragments involved in highly specific recognition, and the binding of a single antigenic determinant or epitope. This is in contrast to polyclonal antibodies that typically include different antibodies directed against different antigenic determinants. The term "monoclonal" antibody or antigen binding fragment thereof refers to both intact and full length monoclonal antibodies, as well as antibody fragments (Fab, Fab', F(ab')2 and Fv), single chain (scFv) variants, antibodies. Included are fusion proteins that include moieties, and any other modified immunoglobulin molecule that includes an antigen recognition site. Furthermore, "monoclonal" antibodies or antigen-binding fragments thereof include any such technique and antigens thereof produced by any number of techniques, including but not limited to hybridoma production, phage selection, recombinant expression, and transfected animals. Refers to the binding fragment.

The term “humanized” antibody or antigen-binding fragment thereof is a non-human (eg, murine) antibody that is a specific immunoglobulin chain, chimeric immunoglobulin, or fragment thereof that contains minimal non-human (eg, murine) sequences. Alternatively, it refers to the form of an antigen-binding fragment. Typically, a humanized antibody or antigen-binding fragment thereof will comprise a non-human species (eg , Mouse, rat, rabbit, hamster), which is a human immunoglobulin substituted with residues derived from CDRs (Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-. 327 (1988); Verhoeyen et al., Science 239:1534-1536 (1988)). In some examples, the Fv framework region (FR) residue of a human immunoglobulin is replaced by the corresponding residue of an antibody or fragment from a non-human species that has the desired specificity, affinity, and potency. To be The humanized antibody or antigen-binding fragment thereof may be further modified by making additional residue substitutions in the Fv framework regions and/or within the replaced non-human residue to provide the antibody or antigen-binding fragment thereof. The specificity, affinity and/or capacity of the fragments can be improved and optimized. Generally, a humanized antibody or antigen-binding fragment thereof comprises at least one, and typically 2 or 3 variable domains, substantially all or substantially all of the CDR regions corresponding to a non-human immunoglobulin. All, or substantially all, of the FR regions are of human immunoglobulin consensus sequences. The humanized antibody or antigen-binding fragment thereof can also include at least a portion of an immunoglobulin constant region or domain (Fc), typically that of a human immunoglobulin. Examples of methods used to generate humanized antibodies are described in US Pat. No. 5,225,539; Roguska et al. , Proc. Natl. Acad. Sei. , USA, 91(3):969-973 (1994), and Roguska et al. , Protein Eng. 9(10):895-904 (1996). In some embodiments, "humanized antibody" is a resurfaced antibody.

The “variable region” of an antibody, alone or in combination, refers to the variable region of an antibody light chain or the variable region of an antibody heavy chain. The variable regions of the heavy and light chains each consist of four framework regions (FR) connected by three complementarity determining regions (CDRs), also known as hypervariable regions. The CDRs in each chain are held together very closely with the FRs and, together with the CDRs from the other chains, contribute to the formation of the antigen-binding site of the antibody. At least two techniques for determining CDRs: (1) Diversity-based approaches to heterologous sequence (ie, Kabat et al., Sequences of Proteins of Immunological Interest, (5th edition, 1991, National Institutes of Health,. Bethesda Md.) (“Kabat”); and (2) an approach based on crystallographic studies of antigen/antibody complexes (Al-lazikani et al., J. Molec. Biol. 273:927-948 (1997). In addition, the art may use a combination of these two approaches to determine CDRs.

The "constant" regions of an antibody are not directly involved in binding the antibody to its antigen, but exhibit various effector functions such as involvement of the antibody in antibody-dependent cellular cytotoxicity.

When referring to residues in the variable domain (approximately residues 1 to 107 of the light chain and residues 1 to 113 of the heavy chain), the Kabat numbering system is generally used (see, eg, Kabat et al. , Sequences of Immunological Interest. 5th Edition., 1991, National Institutes of Health, Bethesda, Md. ("Kabat").

Numbering of amino acid positions as in Kabat is as described in Kabat et al. (Sequences of Immunological Interest. 5th Edition., 1991, National Institutes of Health, Bethesda, Md.) (“Kabat”) used for heavy and light chain variable domains in organizing antibodies. Refers to the numbering system. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids that correspond to contracting or inserting the FRs or CDRs of the variable domain. For example, the heavy chain variable domain can include a single amino acid insert after residue 52 of H2 (residue 52a according to Kabat) and insert a residue after residue 82 of heavy chain FR. Can (residues 82a, 82b and 82c, etc. according to Kabat). The Kabat numbering of residues for a given antibody can be determined by sequence comparisons in the regions of homology of the antibody's sequence with the "standard" Kabat numbering sequence. Chothia instead refers to the position of the loop on the structure (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). The ends of the Chothia CDR-H1 loop, when numbered using the Kabat numbering transformation, vary between H32 and H34 depending on the length of the loop (which is different from Kabat's numbering scheme as H35A). This is because the insertion is placed at H35B, the loop ends at 32 if neither 35A nor 35B is present, and the loop ends at 33 if only 35A is present, since both 35A and 35B are present. If so, the loop ends at 34). The hypervariable region of AbM represents a compromise between the CDRs of Kabat and the structural loop of Chothia and is used by Oxford Molecular's AbM antibody modeling software.

The term “human” antibody or antigen-binding fragment thereof has the amino acid sequence corresponding to the antibody or antigen-binding fragment thereof produced by human, or the antibody or antigen-binding fragment thereof produced by human, and is known in the art. Antibody or antigen-binding fragment thereof produced by any of the techniques described above. This definition of human antibodies or antigen binding fragments thereof includes intact or full length antibodies and fragments thereof.

The term "chimeric" antibody or antigen binding fragment thereof refers to an antibody or antigen binding fragment thereof in which the amino acid sequence is derived from two or more species. Typically, the variable regions of both the light and heavy chains are antibodies or derived from one of the mammalian species (eg, mouse, rat, rabbit, etc.) with the desired specificity, affinity, and potency. The constant region, while corresponding to the variable region of an antigen binding fragment, is homologous to the sequence of an antibody or antigen binding fragment thereof from another species (usually human) and is responsible for eliciting an immune response in that species. To avoid.

The terms "epitope" or "antigenic determinant" are used interchangeably herein and refer to the portion of an antigen that is recognized by a particular antibody and capable of being specifically bound. When the antigen is a polypeptide, the epitope can be formed from both contiguous and noncontiguous amino acids juxtaposed by tertiary folding of the protein. Epitopes formed from contiguous amino acids are typically retained during protein denaturation, whereas epitopes formed by tertiary folding are typically lost during protein denaturation. Be seen. Epitopes typically include at least 3, and more usually, at least 5 or 8-10 amino acids in a unique spatial organization.

“Binding affinity” generally refers to the total strength of non-covalent interactions between a single binding site of a molecule (eg, antibody) and its binding partner (eg, antigen). Unless otherwise indicated, "binding affinity" as used herein refers to a unique binding affinity that reflects a 1:1 interaction between members of a binding pair (eg, antibody and antigen). The affinity of molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Antibodies with low affinity generally bind antigen slowly and tend to dissociate rapidly, whereas antibodies with high affinity generally bind antigen fast and tend to remain bound for a long time. Various methods of measuring binding affinity are known in the art, any of which can be used for the purposes of the present invention.

"Better" as used herein to refer to binding affinity refers to the stronger binding between a molecule and its binding partner. "Better" as used herein refers to stronger binding represented by smaller Kd values. For example, for an antibody having an affinity for the antigen "better than 0.6 nM", the affinity of the antibody for the antigen is less than 0.6 nM, ie 0.59 nM, 0.58 nM, 0.57 nM, etc., or 0. Any value less than 6 nM.

By "specifically binds" is usually meant that the antibody binds to the epitope via its antigen binding domain and that the binding involves some degree of complementarity between the antigen binding domain and the epitope. .. By this definition, an antibody is said to "specifically bind" to an epitope if it binds to the epitope via its antigen binding domain more easily than it binds to a random, unrelated epitope. .. The term "specificity" is used herein to specify the relative affinity of a particular antibody for binding a particular epitope. For example, antibody "A" may be considered to have a higher specificity for a given epitope than antibody "B", or antibody "A" may have a higher specificity for a related epitope "D". It can also be said that it binds to epitope "C" with high specificity.

By "preferentially binds" is meant that the antibody specifically binds to an epitope more easily than it binds to a related, similar, homologous, or similar epitope. Thus, even though antibodies that "preferentially bind" to a given epitope can cross-react with related epitopes, such antibodies are more likely to bind that epitope than related epitopes.

An antibody is said to "competitively inhibit" binding of a reference antibody to a given epitope if it preferentially binds to that epitope or an overlapping epitope to the extent that it blocks binding of the reference antibody to that epitope to some extent. .. Competitive inhibition may be measured by any method known in the art, eg, a competitive ELISA assay. An antibody may be said to competitively inhibit the binding of a reference antibody to a given epitope by at least 90%, at least 80%, at least 70%, at least 60%, or at least 50%.

The phrase “substantially similar” or “substantially identical” as used herein refers to two numerical values (generally one relating to an antibody of the invention and the other to a reference/comparative antibody). Within the context of the biological characteristics measured by (related to), one of ordinary skill in the art would consider that the difference between the two values has little or no biological and/or statistical significance. A significantly higher degree of similarity between the above values (eg Kd values) such that The difference between the two values can be less than about 50%, less than about 40%, less than about 30%, less than about 20%, or less than about 10% as a function of the value of the reference/comparative antibody.

An "isolated" polypeptide, antibody, polynucleotide, vector, cell or composition is a polypeptide, antibody, polynucleotide, vector, cell or composition in a form not found in nature. Isolated polypeptides, antibodies, polynucleotides, vectors, cells or compositions include those that have been purified to the extent that they are no longer in their naturally occurring form. In some embodiments, the isolated antibody, polynucleotide, vector, cell or composition is substantially pure.

As used herein, "substantially pure" means at least 50% pure (ie, contaminant-free), at least 90% pure, at least 95% pure, at least 98% pure or at least 99% pure. Refers to a substance.

The term “immunoconjugate” or “conjugate” as used herein refers to a compound or derivative thereof that is linked to a cell binding agent (ie, an anti-CD33 antibody or antigen binding fragment thereof) and has the general formula: Defined by CLA, where C = cytotoxin, L = linker, and A = anti-CD33 antibody or antibody fragment. Immunoconjugates can also be defined by the general formula in reverse order: ALC.

A "linker" is any chemical compound that is capable of linking a compound, usually a drug such as an indolinobenzodiazepine dimer, to a cell-binding agent such as an anti-CD33 antibody or antigen-binding fragment thereof, in a stable covalent bond. It is a part. The linker may be sensitive to acid-induced cleavage, light-induced cleavage, peptidase-induced cleavage, esterase-induced cleavage, and disulfide bond cleavage under conditions in which the compound or antibody remains active. Can be, or can be substantially resistant. Suitable linkers are well known in the art and include, for example, disulfide groups, and thioether groups.

The term "IMGN779" refers to the huMy9-6 antibody conjugated to DGN462 via the cleavable disulfide linker, N-succinimidyl 4-(2-pyridyldithio)2-sulfobutanoic acid (sulfo-SPDB). Including a CD33-targeted ADC (ie, heavy chain CDRs 1-3 having the sequences SEQ ID NOS: 1-3, respectively, and light chain CDRs 1-3 having the sequences SEQ ID NOS: 4-6; A chain variable region and a light chain variable region having the sequence of SEQ ID NO: 10; an antibody comprising a heavy chain sequence having the sequence of SEQ ID NO: 11 and a light chain sequence having the sequence of SEQ ID NO: 12). The huMy9-6 antibody is also known as the Z4681A antibody. IMGN779 has Formulas IV and V shown below:

Or a pharmaceutically acceptable salt thereof; and

Alternatively, it is formulated as a combination of pharmaceutically acceptable salts thereof.

The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals in which a population of cells is characterized by unregulated cell growth. Examples of cancer include acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute lymphoblastic leukemia (ALL), T cell acute lymphoblastic leukemia (T-ALL), B cell lineage. Acute lymphoblastic leukemia (BALL), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), myelodysplastic syndrome (MDS), basic plasmacytoid DC tumor (BPDCN) leukemia, non Hodgkin lymphoma (NHL), mantle cell lymphoma, and Hodgkin leukemia (HL). The cancer can be a cancer that expresses CD33.

The terms "cancer cell," "tumor cell," and grammatical equivalents refer to a tumor or precancerous lesion that includes both non-tumorigenic cells that comprise the majority of the tumor cell population and tumorigenic stem cells (cancer stem cells). Refers to the entire population of cells derived from. As used herein, the term "tumor cell" is modified by the term "non-tumorigenic" when it distinguishes them from cancer stem cells simply by referring to those tumor cells that lack the ability to regenerate and differentiate. Will be done.

An "advanced" cancer is one that has spread outside the original site or organ, either by local invasion or metastasis. The term "advanced" cancer includes both locally advanced and metastatic disease.

"Metastatic" cancer refers to cancer that has spread from one part of the body to another part of the body.

“Refractory” cancer is one that progresses despite the administration of antitumor treatments such as chemotherapeutic agents to cancer patients.

A "recurrent" cancer is one that has regrown at either an early or distant site after response to initial therapy.

A "relapsed" patient is one that has the signs or symptoms of cancer after the recurrence. Optionally, the patient has relapsed after adjuvant or neoadjuvant therapy.

The term "treatment policy" or "therapy policy" refers to surgery, radiation therapy, chemotherapy, differentiation therapy, biotherapy, immunotherapy, or one or more anti-cancer agents (eg, cytotoxic agents, anti-proliferative compounds, and /Or angiogenesis inhibitors), but not limited thereto.

The terms “first line therapy”, “first line therapy”, and “front line therapy” refer to the preferred and standard initial treatment for a particular condition, eg, a given type and stage of cancer. These treatments are different from the "second-line" therapies that are tried when the first-line therapies do not work well. "Tertiary" therapy is tried when primary and secondary therapies do not work well.

The term "maintenance therapy" refers to therapy given to the cancer to prevent it from recurring after it has been eliminated after the initial therapy.

The term “ineligible AML” as used herein refers to a subject with AML who is not eligible for intensive care. Measurements to determine subjects with inappropriate AML include, for example, physical performance (eg, US East Coast Cancer Clinical Trials Group Performance Status (ECOG PS), Karnofsky Performance Status (KPS), and Brief Physical Fitness Battery). (As determined by (SPPB)), comorbidity (as determined by the Charlson Complication Index Score (CCI) or Hematopoietic Cell Transplantation Specific Comorbidity Index (HCT-CI)), cognitive function, and prognostic model (Including but not limited to cytogenetic groups, age, white blood cell count, LDH, AML types). In some cases, the inappropriate AML subject is a subject over the age of 60.

The term "appropriate AML" as used herein refers to a subject with AML who is eligible for intensive care. Measurements to determine subjects with suitable AML include, for example, physical performance (eg, US East Coast Cancer Clinical Trials Group Performance Status (ECOG PS), Karnofsky Performance Status (KPS), and Brief Physical Performance Battery ( (As determined by SPPB)), comorbidity (as determined by the Charlson Complication Index Score (CCI) or Hematopoietic Cell Transplantation Specific Comorbidity Index (HCT-CI)), cognitive function, and prognostic model ( (Including but not limited to cytogenetic groups, age, white blood cell count, LDH, AML types). In some cases, suitable AML subjects are those who are 60 years of age or older or under 60 years of age.

The term "P-glycoprotein", as used herein, unless otherwise indicated, refers to any naturally occurring P-glycoprotein. The term "P-glycoprotein" includes "full length" unprocessed P-glycoprotein, as well as any form of P-glycoprotein that results from intracellular processing. The term also includes naturally occurring variants of P-glycoprotein, such as those encoded by splice variants and allelic variants. The P-glycoproteins described herein can be isolated from a variety of sources, such as human tissue type or another source, or can be prepared by recombinant or synthetic methods. Unless otherwise specified, "P-glycoprotein" can be used to refer to a nucleic acid that encodes a P-glycoprotein. Human P-glycoprotein sequences are known and include, for example, the publicly available sequences (including isoforms) under NCBI accession number NP_001035830. As used herein, the term “human P-glycoprotein” refers to a P-glycoprotein comprising the sequence of SEQ ID NO:15:

The terms "FLT3 protein", "FLT3 polypeptide", "FLT3", "FLT-3 receptor", or "FLT-3R", as used herein, unless otherwise indicated, refer to any naturally occurring protein. Refers to the FLT3 protein. The term “FLT3” encompasses “full length” untreated FLT3, as well as any form of FLT3 that results from intracellular treatment. The term also includes naturally occurring variants of FLT3, such as those encoded by splice variants and allelic variants. The FLT3 polypeptides described herein can be isolated from a variety of sources, such as human tissue type or another source, or can be prepared by recombinant or synthetic methods. Unless otherwise specified, "FLT3" can be used to refer to a nucleic acid encoding a FLT3 polypeptide. Human FLT3 sequences are known and include, for example, the publicly available sequences (including isoforms) under NCBI accession number NP_004110. As used herein, the term "human FLT3" refers to FLT3 comprising the sequence of SEQ ID NO:16:

By "FLT3-ITD" is meant an FLT3 polypeptide having internal tandem duplication(s), including but not limited to simple tandem duplication(s) and/or tandem duplication(s) with insertions. .. In various embodiments, the FLT3 polypeptide with internal tandem duplication is an activated FLT3 variant (eg, constitutively autophosphorylated). In some embodiments, the FLT3-ITD includes tandem duplications in exons or introns, including, for example, exon 11, exon 11-intron 11, and exon 12, exon 14, exon 14-intron 14, and exon 15. And/or tandem duplication(s) with insertions. The internal tandem duplication mutation (FLT3-ITD) is the most common FLT3 mutation present in approximately 20-25% of AML cases. FLT3-ITD AML patients have a worse prognosis than wild-type (WT) FLT3 patients, with high recurrence rates and short-duration response to chemotherapy.

By "analog" is meant a molecule that is not identical, but has similar functional or structural characteristics. For example, a polypeptide analog has certain biochemical modifications that retain the biological activity of the corresponding naturally occurring polypeptide while enhancing the function of the analog as compared to the naturally occurring polypeptide. .. Such biochemical modifications can, for example, increase the protease resistance, membrane permeability, or half-life of the analog without altering ligand binding. Analogs may include unnatural amino acids.

The term “subject” refers to any animal (eg, mammal) including, but not limited to, humans, non-human primates, rodents, etc., which are recipients of a particular treatment. Generally, the terms "subject" and "patient" are used interchangeably herein with reference to a human subject.

The term "pharmaceutical formulation" refers to a preparation that is in a form that renders the biological activity of the active ingredient effective and does not contain additional ingredients that are unacceptably toxic to the subject to which the formulation is administered. The formulation may be sterile.

An “effective amount” of an antibody, immunoconjugate, or other drug as disclosed herein is an amount sufficient to carry out a specifically mentioned purpose.

The term "therapeutically effective amount" refers to the amount of antibody, immunoconjugate, or other drug effective to "treat" a disease or disorder in a subject or mammal. In the case of cancer, a therapeutically effective amount of the drug can reduce the number of cancer cells; reduce the size or burden of the tumor; inhibit the invasion of cancer cells into peripheral organs (ie Can be delayed, to some extent, can be stopped in certain embodiments); tumor metastasis can be inhibited (ie, delayed to some extent, stopped in certain embodiments); tumor growth can be inhibited to some extent One or more of the symptoms associated with cancer can be alleviated to some extent; and/or increased progression-free survival (PFS), disease-free survival (DFS) or overall survival (OS), complete An advantageous response such as a response (CR), partial response (PR) or possibly stable disease (SD), reduced progressive disease (PD), reduced time to progression (TTP), or any combination thereof. Can bring. See the definition herein of "treating". To the extent the drug interferes with growth and/or kills existing cancer cells, the drug may be cytostatic and/or cytotoxic.

The term “responsively beneficial” generally refers to producing a beneficial condition in a subject. With respect to treating cancer, the term refers to providing a therapeutic effect to a subject. A favorable therapeutic effect in cancer can be measured by a number of methods (see WA Weber, J. Nucl. Med. 50:1S-10S (2009)). For example, inhibition of tumor growth, expression of molecular markers, expression of serum markers and molecular imaging methods can all be used to assess the therapeutic efficacy of anti-cancer therapy. For inhibition of tumor growth, T/C≦42% is the minimum level of antitumor activity according to NCI criteria. A T/C of less than 10% is considered a high level of anti-tumor activity, T/C (%) = median tumor volume with treatment/median tumor volume of control x 100. An advantageous response may be, for example, increased progression-free survival (PFS), recurrence-free survival (EFS), disease-free survival (DFS), or overall survival (OS), complete response (CR), partial response (PR). ), or optionally stable disease (SD), progressive disease (PD) reduction, reduced time to progression (TTP), or any combination thereof.

PFS, DFS and OS can be measured according to the standards set by the National Cancer Institute and the US Food and Drug Administration for new drug approval. Johnson et al, (2003)J. Clin. Oncol. 21(7):1404-1411.

Progression-free survival (PFS) refers to the time from enrollment to disease progression or death. PFS is generally measured using the Kaplan-Meier method and the Response Evaluation Criteria in Solid Tumors (RECIST) 1.1 standard. Generally, progression-free survival refers to the situation in which a patient remains alive without the cancer getting worse.

"Relapse-free survival" (EFS) refers to the length of time, without complications or events, intended for a patient to prevent or delay treatment after the end of primary treatment. The event may include the recurrence of cancer, or the development of certain symptoms.

"Time to tumor progression" (TTP) is defined as the time from enrollment to disease progression. TTP is generally measured using the RECIST 1.1 standard.

"Complete response" or "complete response" or "CR" indicates that all signs of tumor or cancer are shed in response to treatment. This does not necessarily mean that the cancer has been cured.

"Complete remission with inadequate restoration of hematopoietic function" or "CRi" means that the number of blast cells in the bone marrow is less than 5%, but the blood cell count (for example, neutrophil and platelet) is not within the normal range Refers to the patient response.

"Partial response" or "PR" refers to a reduction in the size or volume of one or more tumors or lesions, or a reduction in the degree of cancer in the body in response to treatment.

“Stable disease” refers to a disease that does not progress or relapse. In stable disease, there is neither sufficient tumor atrophy to be considered a partial response nor sufficient tumor growth to be considered a progressive disease.

"Progressive disease" refers to the appearance of another new lesion or tumor and/or the definite progression of an existing non-target lesion. Progressive disease can also refer to the growth of tumors by more than 20% since the beginning of treatment, either by increasing the mass or increasing the spread of the tumor.

“Disease-free survival (DFS)” refers to the length of time that a patient remains disease-free during and after treatment.

"Overall Survival (OS)" refers to the time from patient enrollment to the time of censorship at the time of death or last known survival. OS includes prolonging life expectancy as compared to naive or untreated individuals or patients. Overall survival refers to the situation in which a patient remains alive for a defined period of time, eg, 1 year, 5 years, etc., from the time of diagnosis or treatment. In a population of patients, overall survival is measured as the mean overall survival (mOS).

Terms such as "treating" or "treatment" or "to treat" or "alleviating" or "to alleviate" have been diagnosed Refers to a therapeutic means that cures, slows down, reduces, and/or halts the progression of the symptoms of the condition or disorder. Thus, those in need of treatment include those already diagnosed with the disorder or suspected of having the disorder. In certain embodiments, the patient has the following: reduced number of cancer cells or complete absence of cancer cells; reduced tumor size; eg, cancer to peripheral organs, including cancer spread to soft tissue and bone. Inhibition or absence of cell invasion; Inhibition or absence of tumor metastasis; Inhibition or absence of tumor growth; Alleviation of one or more symptoms associated with a particular cancer; Reduction of morbidity and mortality; Improved quality; decreased tumorigenicity, tumorigenicity frequency or tumorigenic capacity of the tumor; decreased number or frequency of cancer stem cells in the tumor; differentiation of tumorigenic cells to a non-tumorigenic state; increased absence Exacerbation Survival (PFS), Disease Free Survival (DFS) or Overall Survival (OS), Complete Response (CR), Partial Response (PR), Stable Disease (SD), Progressive Disease (PD) Reduction, Until Progression Subject has been successfully “treated” for cancer according to the methods of the present invention if it exhibits one or more of the following: a reduction in time (TTP), or a combination thereof.

The terms “administering”, “administering”, “administration” and the like, as used herein, refer to the method used to allow delivery of the immunoconjugate to the desired site of biological action. Point to. Administration techniques that can be used in the agents and methods described herein are described, for example, in Goodman and Gilman, The Pharmacologic Basis of Therapeutics, current ed. Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co.; , Easton, Pa. In one aspect, the immunoconjugate is administered intravenously.

The term "instructing" provides instructions for any applicable therapy, dosing, treatment, treatment regimen, etc., by any means, eg, in writing, eg, in the form of a package insert or other written promotional material. Means that.

The terms “polypeptide”, “peptide”, and “protein” are used interchangeably herein to refer to polymers of amino acids of any length. The polymer can be linear or branched, it can comprise modified amino acids, and it can be interrupted by non-amino acids. The term is modified naturally or is modified by intervention, such as disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation or any other manipulation or modification, such as conjugation with a labeling moiety. Included amino acid polymers. Also included within the definition are, for example, polypeptides containing one or more analogs of an amino acid (including, for example, unnatural amino acids, etc.) as well as other modifications known in the art. Since the polypeptides of the present invention are antibody-based, it is understood that in certain embodiments, the polypeptides may exist as single chains or associated chains.

The term "identical" or percent "identity" in the context of two or more nucleic acids or polypeptides is compared for maximum correspondence without consideration of conservative amino acid substitutions as part of sequence identity, (necessary When aligned (introducing a gap accordingly), it refers to two or more sequences or subsequences that are the same, or two or more sequences or subsequences that have the specified percentage of nucleotide or amino acid residues that are the same. Percent identity can be measured using sequence comparison software or algorithms, or by visual inspection. Various algorithms and software are known in the art that can be used to obtain sequence comparisons of amino acid or nucleotide sequences. One such non-limiting example of a sequence comparison algorithm is described by Karlin et al. , Proc. Natl. Acad. Sci. , 87:2264-2268 (1990), and Karlin et al. , Proc. Natl. Acad. Sci. , 90:5873-5877 (1993) and incorporated into the NBLAST and XBLAST programs (Altschul et al., Nucleic Acids Res., 25:3389-3402 (1991)). In certain embodiments, gapped BLAST can be prepared as described in Altschul et al. , Nucleic Acids Res. 25:3389-3402 (1997). BLAST-2, WU-BLAST-2 (Altschul et al., Methods in Enzymology, 266:460-480 (1996)), ALIGN, ALIGN-2 (Genentech, South San Francisco, Calif.), or Meg. There are additional publicly available software programs that can be used to align the sequences. In certain embodiments, the percent identity between two nucleotide sequences is determined by the GCG software (eg, NWSgapdna.CMP index and 40, 50, 60, 70, or 90 gap weights and 1, 2, 3, 4, 5 or 6 (with a length weight of 6). In certain alternative embodiments, a GAP program in the GCG software package that incorporates the algorithm of Needleman and Wunsch (J. Mol. Biol. (48):444-453 (1970)) is used (eg, the Blossum 62 indicator or PAM 250. Determining the percent identity between two amino acid sequences (using indicators and gap weights of 16, 14, 12, 10, 8, 6, or 4 and length weights of 1, 2, 3, 4, 5). You can Alternatively, in certain embodiments, the percent identity between nucleotide or amino acid sequences is determined using the algorithm of Myers and Miller (CABIOS, 4:11-17 (1989)). For example, percent identity can be determined using the ALIGN program (version 2.0) and using PAM120 with a residue table, a gap length penalty of 12, and a gap penalty of 4. Appropriate parameters for maximum sequence comparisons with particular sequence comparison software can be determined by one of ordinary skill in the art. In certain embodiments, the default parameters of sequence comparison software are used. In certain embodiments, the percent identity “X” of a first amino acid sequence to a second sequence amino acid is calculated as 100×(Y/Z), where Y is the first and second The number of amino acid residues scored as the same match in a sequence comparison (aligned by visual inspection or a particular sequence comparison program) of the sequences, and Z is the total number of residues in the second sequence. If the length of the first sequence is longer than the second sequence, then the percent identity of the first sequence to the second sequence is greater than the percent identity of the second sequence to the first sequence. Would be

By way of non-limiting example, any particular polynucleotide has a certain ratio of sequence identity to a reference sequence (eg, at least 80% identical, at least 85% identical, at least 90% identical). , And in some embodiments at least 95%, 96%, 97%, 98% or 99% identical), in certain embodiments, the Bestfit program (Wisconsin Sequence Analysis Package, Unix®). Version 8, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, WI 53711). Bestfit uses the local homology algorithm of Smith and Waterman (Advances in Applied Mathematics 2:482, 489 (1981)) to find the best segment of homology between two sequences. When using Bestfit or any other sequence comparison program to determine whether a particular sequence is, for example, 95 percent identical to a reference sequence according to the present invention, the parameter is the percentage of identity referred to. Calculated over the full length of the nucleotide sequence and set to allow gaps in homology up to 5% of the total number of nucleotides in the reference sequence.

In some embodiments, the two nucleic acids or polynucleotides of the invention are substantially identical, which compares and aligns for the largest match, as measured using a sequence comparison algorithm or by visual inspection. They are at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, and in some embodiments at least 95%, 96%, 97%, 98%, 99%. It is meant to have nucleotide or amino acid residue identity. Identity is over a region of the sequence that is at least about 10, about 20, about 40-60 residues in length, or an integer value in between, or is greater than 60-80 residues, eg, at least about 90-100 residues. Present over a region, and in some embodiments, the sequences are substantially identical over the full length of the sequences being compared, eg, the coding regions of the nucleotide sequences.

A "conservative amino acid substitution" is one in which one amino acid residue is replaced with another amino acid residue having a similar side chain. Basic side chains (eg lysine, arginine, histidine), acidic side chains (eg aspartic acid, glutamic acid), uncharged polar side chains (eg glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), non Polar side chains (eg, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), β-branched side chains (eg, threonine, valine, isoleucine) and aromatic side chains (eg, tyrosine, phenylalanine, tryptophan, A family of amino acid residues with similar side chains, including histidine) has been defined in the art. For example, the substitution of tyrosine for phenylalanine is a conservative substitution. In some embodiments, conservative substitutions in the sequences of the polypeptides and antibodies of the present invention will bind to the antigen(s) of the polypeptide or antibody containing the amino acid sequence, ie, the polypeptide or antibody will bind. Does not abolish binding to CD33. Methods for identifying conservative substitutions of nucleotides and amino acids that do not eliminate antigen binding are well known in the art (eg, Brummell et al., Biochem. 32:1180-1 187 (1993); Kobayashi et al.,. Protein Eng. 12(10):879-884 (1999); and Burks et al., Proc. Natl. Acad. Sci. USA 94:.412-417 (1997)).

Unless specifically stated or clear from the description, the term "about" as used herein is understood to be included within the general tolerances in the art, eg, within 2 standard deviations of the mean. To be done. About 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05% of the standard value , Or within 0.01%. Unless otherwise apparent from the written description, all numerical values provided herein are modified by the term about.

The recitation of a listing of chemical groups in any definition of a variable herein includes the definition of that variable as any single group or combination of listed groups. A description of an embodiment with respect to a variable part or aspect herein includes that embodiment as any single embodiment or in combination with any other embodiment or part thereof.

As used in this disclosure and the claims, the singular forms "a", "an", and "the" include plural forms unless the context clearly dictates otherwise.

Wherever an embodiment is described herein with the language "comprising", the phrase "consisting of" and/or "consisting essentially of". It is understood that another similar embodiment as described in 1. is also provided. In this disclosure, "comprises," "comprising," "containing," and "having" and the like are given to them in the United States Patent Act. It can have meaning and can mean "includes," "includes," etc.; "consisting essentially of" and "consistently essentially". Has the meaning given in US patent law, which terms are open-ended and whose basic or novel features are listed more than those listed. The presence of more than that listed is permitted, except for prior art embodiments, unless the presence of something alters it.

Unless specifically stated or clear from the description, the term "or" as used herein is understood to be inclusive. The term "and/or" as used herein with terms such as "A and/or B" includes "A and B," "A or B," "A" and "B." Is intended to include. Similarly, the term "and/or" when used in a phrase such as "A, B and/or C" has the following embodiments: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone), respectively.

Any composition or method provided herein can be combined with one or more of any of the other compositions and methods provided herein.

II. Anti-CD33 Antibodies and Antigen Binding Fragments Thereof Described herein are methods of administering an immunoconjugate that specifically binds to CD33 (eg, IMGN779). The immunoconjugate comprises an anti-CD33 antibody or antigen binding fragment thereof.

The anti-CD33 antibody can be a huMy9-6 antibody.

"My9-6" (also known as "murine My9-6" and "muMy9-6") is a murine anti-CD33 antibody derived from huMy9-6. My9-6 has been fully characterized for germline amino acid sequences of both light and heavy chain variable regions, amino acid sequences of both light and heavy chain variable regions, CDR identification, surface amino acid identification. And means its expression in recombinant form. For example, US Pat. Nos. 7,557,189; 7,342,110; 8,119,787; 8; the contents of each of which are incorporated herein by reference in their entirety. , 337,855 and US Patent Publication No. 20120244171. The hybridoma producing the muMy9-6 antibody was deposited with the American Type Culture Collection (PO Box 1549, Manassas, Va. 20108) on November 7, 2002 under the terms of the Budapest Treaty, and the accession number is PTA-4786. Was given. The amino acid sequence of muMy9-6 is also shown in Table 1 below. The My9-6 antibody has also been functionally characterized and shown to bind CD33 with high affinity on the surface of CD33-positive cells.

The humanized version of My9-6 is referred to herein as "huMy9-6" or "humanized My9-6". The CDR sequences of huMy9-6 are also provided in Table 1. To generate hyMy9-6, CDRs of My9-6 were identified by modeling and predicted their molecular structure. Humanized antibodies were then prepared and fully characterized, eg, as described in US Pat. Nos. 7,342,110 and 7,557,189, incorporated herein by reference. .. The amino acid sequences of the light and heavy chains of many huMy9-6 antibodies are found, for example, in US Pat. No. 8,337,855 and US Pat. No. 8,765,740, each of which is incorporated herein by reference. It is described in. The variable heavy and light chain amino acid sequences of huMy9-6, and the heavy and light chain amino acid sequences are provided in Table 2.

In some embodiments, the anti-CD33 immunoconjugate (eg, IMGN779) comprises a humanized antibody or antigen binding fragment thereof. In some embodiments, the humanized antibody or fragment is a resurfaced antibody or antigen binding fragment thereof. In other embodiments, the anti-CD33 immunoconjugate (eg, IMGN779) comprises a fully human antibody or antigen binding fragment thereof.

In some embodiments, the anti-CD33 antibody or antigen-binding fragment comprises the heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 1-3, respectively, and the light chain CDR1, CDR2 set forth in SEQ ID NOs: 4-6, respectively. , And CDR3 sequences.

In some embodiments, the anti-CD33 antibody or antigen binding fragment comprises the heavy chain variable domain sequence of SEQ ID NO:9. In some embodiments, the anti-CD33 antibody or antigen binding fragment comprises the light chain variable domain sequence of SEQ ID NO:10. In some embodiments, the anti-CD33 antibody or antigen binding fragment comprises the heavy chain variable domain sequence of SEQ ID NO:9 and the light chain variable domain of SEQ ID NO:10.

In some embodiments, the anti-CD33 antibody or antigen-binding fragment comprises the heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 1-3, respectively, and the light chain CDR1, CDR2 set forth in SEQ ID NOs: 4-6, respectively. , And the CDR3 sequence, and the heavy chain variable domain sequence of SEQ ID NO:9. In some embodiments, the anti-CD33 antibody or antigen-binding fragment comprises the heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 1-3, respectively, and the light chain CDR1, CDR2 set forth in SEQ ID NOs: 4-6, respectively. , And the CDR3 sequence, and the light chain variable domain sequence of SEQ ID NO:10.

In some embodiments, the anti-CD33 antibody or antigen binding fragment thereof comprises the heavy chain sequence of SEQ ID NO:11 and the light chain sequence of SEQ ID NO:12.

In a further embodiment, the anti-CD33 antibody or antigen binding fragment thereof comprises at least one heavy chain variable region, and at least one light chain variable region, wherein said heavy chain variable regions are represented by SEQ ID NOs: 1-3, respectively. And the light chain variable region has three amino acid sequences represented by SEQ ID NOs: 4 to 6, respectively.

In a further embodiment, the anti-CD33 antibody or antigen binding fragment thereof is a humanized antibody or antigen binding fragment thereof comprising at least one heavy chain variable region and at least one light chain variable region, wherein the heavy chain variable region is , Three complementarity determining regions each having an amino acid sequence represented by SEQ ID NOS: 1-3, wherein the light chain variable region has three complementarity determining regions each having an amino acid sequence represented by SEQ ID NOS: 4-6. Including the area.

In a further embodiment, the anti-CD33 antibody or antigen binding fragment thereof is a CDR-grafted or resurfaced antibody comprising at least one heavy chain variable region and at least one light chain variable region, said heavy chain variable region comprising the sequence The light chain variable region comprises three complementarity determining regions each having an amino acid sequence represented by SEQ ID NOS: 1 to 3, and the light chain variable region includes three complementarity determining regions each having an amino acid sequence represented by SEQ ID NOS: 4 to 6. Including.

In a further embodiment there is provided an antibody or antigen binding fragment thereof comprising at least one heavy chain variable region and at least one light chain variable region, wherein said heavy chain variable region is according to SEQ ID NOs: 1, 13 and 3, respectively. The light chain variable region comprises three complementarity determining regions having the amino acid sequences represented by SEQ ID NOs: 4 to 6, respectively.

In a further embodiment, at least 90% sequence identity with the amino acid sequence represented by SEQ ID NO: 9, more preferably with SEQ ID NO: 9 and 95% sequence identity, most preferably with SEQ ID NO: 9 and 100% sequence identity. Antibodies or antigen-binding fragments having humanized (eg, resurfacing, CDR grafted) heavy chain variable regions that share identity are provided. In certain embodiments, the antibody comprises conservative mutations in framework regions outside of the CDRs.

Similarly, at least 90% sequence identity with the amino acid sequence corresponding to SEQ ID NO: 10, more preferably with SEQ ID NO: 10 and 95% sequence identity, most preferably with SEQ ID NO: 10 and 100% sequence identity. Antibodies with shared humanized (eg, resurfacing, CDR grafted) light chain variable regions are provided. In certain embodiments, the antibody comprises conservative mutations in framework regions outside of the CDRs.

Antibodies and antigen-binding fragments of immunoconjugates can include polypeptides that are recombinant polypeptides, natural polypeptides, or synthetic polypeptides.

The polypeptide can be further modified to include additional chemical moieties that are not normally part of the protein. These derivatized moieties may improve protein solubility, biological half-life, or absorption. The moieties may further reduce or eliminate any desired side effects such as proteins. An overview of those parts is given in REMINGTON'S PHARMACEUTICAL SCIENCES, 20th ed. Mack Publishing Co. , Easton, PA (2000).

Methods known in the art for purifying antibodies and other proteins are described in US Patent Publication Nos. 2008/0312425, 2008/0177048, the contents of each of which is incorporated herein by reference in its entirety. , And those described in JP-A-2009/0187005.

III. Anti-CD33 Immunoconjugate In addition to an anti-CD33 antibody or antigen-binding fragment thereof, an anti-CD33 immunoconjugate (eg, IMGN779) has at least one cell linked to the antibody or antigen-binding by a linker (eg, sulfo-SPDB). Includes toxins (eg, DGN462). As used herein, expression "linked to an anti-CD33 antibody or antigen binding fragment thereof" refers to at least one cell linked to the anti-CD33 antibody or antigen binding fragment thereof via a suitable linking group or precursor thereof. Refers to an immunoconjugate containing a toxin derivative.

The cytotoxin can be a benzodiazepine dimer. The cytotoxin can be an indolinobenzodiazepine dimer. The cytotoxin can be a DNA alkylating agent. The cytotoxin can be DGN462.

Immunoconjugates can be prepared by using a linking group to link the cytotoxin to the anti-CD33 antibody or antigen binding fragment thereof. Suitable linking groups are well known in the art and include, for example, disulfide groups, thioether groups, acid labile groups, photolabile groups, peptidase labile groups, and esterase labile groups.

The cytotoxin can be linked to the anti-CD33 antibody or antigen binding fragment thereof, for example, via a disulfide bond. The linker molecule or crosslinker contains reactive chemical groups capable of reacting with the anti-CD33 antibody or antigen binding fragment thereof. Reactive chemical groups for reacting with the cell binding agent can be, for example, N-succinimidyl ester and N-sulfosuccinimidyl ester. In addition, the linker molecule contains a reactive chemical group such as a dithiopyridyl group that can react with the drug to form a disulfide bond. Linker molecules include, for example, N-succinimidyl 4-(2-pyridyldithio)2-sulfobutanoic acid (sulfo-SPDB) (see US Publication No. 20090274713, which is incorporated herein by reference in its entirety). .. For example, since the anti-CD33 antibody or antigen-binding fragment thereof can be modified with a cross-linking reagent, the derived anti-CD33 antibody or antigen-binding fragment thereof containing a free or protected thiol group can then be disulfide-containing or thiol-containing. Reacting with the contained indolinobenzodiazepine dimer to produce the immunoconjugate. Immunoconjugates can be purified by chromatography, including but not limited to HPLC, size exclusion, adsorption, ion exchange and affinity capture, dialysis or tangential flow filtration.

In another aspect of the invention, the anti-CD33 antibody or antigen-binding fragment thereof is linked to a cytotoxic drug via a disulfide bond and a polyethylene glycol spacer in enhancing the potency, solubility, or efficacy of the immunoconjugate. It is connected. Such cleavable hydrophilic linkers are described, for example, in WO2009/0134976, which is incorporated herein by reference in its entirety. An additional benefit of this linker design is the desired high monomer ratio and minimal aggregation of antibody drug conjugates. Is being specifically contemplated in this embodiment, has a polyethylene glycol spacer _{((CH 2 CH 2 O)} n = 1~14), drug loading is a narrow range of 2-8, a disulfide group (-S -S-) is a conjugate of a cell binding agent and a drug linked together. These conjugates exhibit relatively high and potent bioactivity against cancer cells, possess the desired biochemical properties of minimal protein aggregation, high binding yields and high monomer ratios.

In some embodiments, the linker is a linker that includes at least one charged group, eg, as described in US Patent Publication No. 2012/0282282, the contents of which are incorporated herein by reference in its entirety. .. In some embodiments, the charged or procharged crosslinker is a crosslinker that includes a sulfonate, phosphate, carboxyl or quaternary amine substituent, which is modified cell binding agent and cell binding agent-drug conjugate. Solubility is significantly increased, especially for 2-20 drug/antibody linked monoclonal antibody-drug conjugates. A conjugate prepared from a linker containing a procharged moiety will produce one or more charged moieties after the conjugate is metabolized intracellularly. In some embodiments, the linker is N-succinimidyl 4-(2-pyridyldithio)-2-sulfobutanoic acid (sulfo-SPDB).

The immunoconjugate has the formula (I):

Immunoconjugate or a pharmaceutically acceptable salt thereof. Double line between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen, when it is a single bond, X is hydrogen and Y is a -SO ₃ H. The term "A" refers to an anti-CD33 antibody, ie, a heavy chain variable region (VH) complementarity determining region (CDR) 1 sequence of SEQ ID NO:1, a VH CDR2 sequence of SEQ ID NO:2, and a VH CDR3 sequence of SEQ ID NO:3, And a light chain variable region (VL) CDR1 sequence of SEQ ID NO:4, a VL CDR2 sequence of SEQ ID NO:5, and a VL CDR3 sequence of SEQ ID NO:6, which is an antibody or an antigen-binding fragment thereof that specifically binds to CD33. The term "r" is an integer from 1-10.

The immunoconjugates of formula (II) and (III) and their pharmaceutically acceptable salts are specific examples of immunoconjugates that can be used in the therapeutic methods of this disclosure.


“A” and “r” are as defined in formula (I). The term "r" is an integer from 1-10. Methods for preparing immunoconjugates of Formulas II and III (eg, IMGN779) are described in US Pat. Provided by.

In certain embodiments, the antibody portion of the immunoconjugate of formula (I), (II), or (III) is the huMy9-6 antibody, also referred to as "Z4681A". In a specific embodiment, the anti-CD33 immunoconjugate is IMGN779. IMGN779 comprises huMy9-6 or Z4681A antibodies conjugated to DGN462 via the cleavable disulfide linker N-succinimidyl 4-(2-pyridyldithio)-2-sulfobutanoic acid (sulfo-SPDB). .. IMGN779 has Formulas IV and V shown below:

Or a pharmaceutically acceptable salt thereof; and

Alternatively, it is formulated as a combination of pharmaceutically acceptable salts thereof.

In certain embodiments, the immunoconjugate comprises 1 to 10 cytotoxic benzodiazepine dimer compounds, 2 to 9 cytotoxic benzodiazepine dimer compounds, 3 to 8 cytotoxic benzodiazepine dimer compounds. 4 to 7 cytotoxic benzodiazepine dimer compounds or 5 to 6 cytotoxic benzodiazepine dimer compounds.

In certain embodiments, the immunoconjugate is 1-10 indolinobenzodiazepine dimer compounds, 2-9 indolinobenzodiazepine dimer compounds, 3-8 indolinobenzodiazepine dimer compounds, 4-8. 7 indolinobenzodiazepine dimer compounds, or 5 to 6 indolinobenzodiazepine dimer compounds.

In certain embodiments, the immunoconjugate comprises 1-10 DGN462, 2-9 DGN462, 3-8 DGN462, 4-7 DGN462, or 5-6 DGN462.

In certain embodiments, a composition comprising an immunoconjugate described herein comprises at least two immunoconjugates having an average of 1-10 cytotoxic benzodiazepine dimer molecules per antibody or antigen binding fragment thereof. It may include a gate. The average ratio of cytotoxic benzodiazepine dimer molecules per antibody molecule is referred to herein as the drug antibody ratio (DAR). In one embodiment, the DAR is 2-8, 3-7, 3-5, or 2.5-3.5.

Further, in some embodiments, the indolinobenzodiazepine dimer (eg, DGN462) is attached to the anti-CD33 antibody or antigen-binding fragment thereof (eg, sulfo-, via the lysine residue of the antibody or antigen-binding fragment thereof). (By SPDB). In some embodiments, 1-10 indolinobenzodiazepine dimers (eg, DGN462) are conjugated to the anti-CD33 antibody or antigen-binding fragment thereof via 1-10 lysine residues of the antibody or antigen-binding fragment thereof. (Eg, by sulfo-SPDB). In some embodiments, the 2-8 indolinobenzodiazepine dimer (eg, DGN462) is conjugated to the anti-CD33 antibody or antigen-binding fragment thereof via the 2-8 lysine residues of the antibody or antigen-binding fragment thereof. (Eg, by sulfo-SPDB). In some embodiments, the 2-5 indolinobenzodiazepine dimer (eg, DGN462) is conjugated to the anti-CD33 antibody or antigen-binding fragment thereof via 2-5 lysine residues of the antibody or antigen-binding fragment thereof. (Eg, by sulfo-SPDB). In some embodiments, the 3-4 indolinobenzodiazepine dimer (eg, DGN462) is conjugated to the anti-CD33 antibody or antigen-binding fragment thereof via the 3-4 lysine residues of the antibody or antigen-binding fragment thereof. (Eg, by sulfo-SPDB).

The cytotoxic benzodiazepine dimer compounds and conjugates described herein are disclosed, for example, in US Pat. No. 8,765,740, paragraphs [0395] to [0397] and [0598] to [0607], FIG. 1, 15, 22, 23, 38-41, 43, 48, 55 and 60, and Examples 1, 6, 12, 13, 20, 21, 22, 23, 26-30 and 32, and U.S. Pat. , 353, 127, paragraphs [0007] to [0105], [0197] to [0291], FIGS. 1 to 11, 16, 28 and Examples 1 to 7, 9 to 13, 15 and 16, but Can be prepared according to the methods described in US Pat. Nos. 8,765,740 and 9,353,127, which are not limited thereto.

The term "cation" refers to an ion that has a positive charge. The cation can be monovalent (eg, Na ⁺ , K ^+, etc.), divalent (eg, Ca ²⁺ , Mg ^2+, etc.) or polyvalent (eg, Al ^3+, etc.). Preferably the cation is monovalent.

The phrase "pharmaceutically acceptable" indicates that the substance or composition must be chemically and/or toxicologically compatible with the other ingredients, including the formulation, and/or the mammal treated with it. ..

The phrase "pharmaceutically acceptable salt," as used herein, refers to pharmaceutically acceptable organic or inorganic salts of a compound of the invention. Examples of salts include sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate. , Lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentianate, fumarate , Gluconate, glucuronate, sucrose, formate, benzoate, glutamate, methanesulfonate, "mesylate", ethanesulfonate, benzenesulfonate, p-toluenesulfonate , Pamoate salts (ie 1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts, alkali metal (eg sodium and potassium) salts, alkaline earth metal (eg magnesium) salts, And ammonium salts, but are not limited thereto. Pharmaceutically acceptable salts may involve the inclusion of another molecule such as, for example, acetate, succinate or other counterion. The counterion may be an organic or inorganic moiety that stabilizes the charge on the parent compound. Furthermore, a pharmaceutically acceptable salt may have more than one charged atom in its structure. Instances where multiple charged atoms are part of the pharmaceutically acceptable salt can have multiple counterions. Thus, a pharmaceutically acceptable salt can have one or more charged atoms and/or one or more counterion. In certain embodiments, the pharmaceutically acceptable salt is a sodium or potassium salt.

If the compound of the invention is a base, the desired pharmaceutically acceptable salt can be prepared by any suitable method available in the art, eg, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfone. Acids, with inorganic acids such as phosphoric acid, or for example, acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranosidyl acid, such as glucurone. Acids or galacturonic acids, alpha hydroxy acids such as citric acid or tartaric acid, amino acids such as aspartic acid or glutamic acid, aromatic acids such as benzoic acid or cinnamic acid, sulfonic acids such as p-toluenesulfonic acid or ethanesulfone. It may be prepared by treatment of the free base with an organic acid such as an acid.

If the compound of the invention is an acid, the desired pharmaceutically acceptable salt can be prepared by any suitable method, such as amine (primary, secondary or tertiary), alkali metal hydroxide or alkaline earth. It may be prepared by treatment of the free acid with an inorganic or organic base such as a metal hydroxide. Illustrative examples of suitable salts are derived from amino acids such as glycine and arginine, ammonia, primary, secondary and tertiary amines, and cyclic amines such as piperidine, morpholine and piperazine. Organic salts and inorganic salts derived from, but not limited to, sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.

IV. Treatment of Cancer with Anti-CD33 Immunoconjugate The present invention treats a patient with cancer, particularly blood cancer, eg, AML, by administering an anti-CD33 immunoconjugate (eg, IMGN779). Provide a way. In some embodiments, the patient is an inappropriate AML patient. In some embodiments, the patient is a suitable AML patient. As used herein, "hematological cancer" is a cancer that begins in hematopoietic tissue, such as bone marrow, or in cells of the immune system. Examples of hematological cancers are leukemia, lymphoma, and multiple myeloma.

Cancers that can be treated using the methods of the present disclosure include leukemia, lymphoma, and myeloma. The cancer may be chemosensitive; or the cancer may be chemoresistant. More specifically, cancers that can be treated using the methods of the present disclosure include acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), chronic. Myeloid leukemia (CML), acute promyelocytic leukemia (APL), myelodysplastic syndrome (MDS), acute monocytic leukemia (AMOL), hairy cell leukemia (HCL), T cell prolymphocytic leukemia (T-PLL), large granular lymphocytic leukemia, adult T-cell leukemia, small lymphocytic lymphoma (SLL), Hodgkin lymphoma (nodular sclerosis, mixed cell type, lymphoid rich type, lymphopenic type or non- Reduced and nodular lymphocyte-predominant Hodgkin lymphoma), non-Hodgkin's lymphoma (all subtypes), chronic lymphocytic leukemia/small lymphocytic lymphoma, B cell prolymphocytic leukemia, lymphoplasmacytic lymphoma (eg, , Waldenstrom macroglobulinemia), splenic marginal zone lymphoma, plasma cell tumor (plasma cell myeloma, plasmacytoma, monoclonal immunoglobulin deposition, heavy chain disease), extranodal marginal zone B cell lymphoma ( MALT lymphoma), nodular marginal zone B cell lymphoma (NMZL), follicular lymphoma, mantle cell lymphoma, diffuse B cell lymphoma, mediastinal (thymus) large B cell lymphoma, intravascular large B cell lymphoma, primary body cavity Lymphoma, Burkitt lymphoma/leukemia, T-cell prolymphocytic leukemia, T-cell large-granular lymphocytic leukemia, aggressive NK-cell leukemia, adult T-cell leukemia/lymphoma, extranodal NK/T-cell lymphoma (nasal type) , Enteropathy-type T cell lymphoma, hepatosplenic T cell lymphoma, blast type NK cell lymphoma, mycosis fungoides/Sezary syndrome, primary cutaneous CD30-positive T cell lymphoproliferative disease, primary cutaneous undifferentiated large cell lymphoma, Includes lymphomatoid papulosis, angioimmunoblastic T cell lymphoma, peripheral T cell lymphoma (unspecified), anaplastic large cell lymphoma), and multiple myeloma (plasma cell myeloma Koehler disease).

In another embodiment, the cancer is acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute lymphoblastic leukemia (ALL), T cell acute lymphoblastic leukemia (T-ALL), B-cell lineage of acute lymphoblastic leukemia (BALL), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), myelodysplastic syndrome, basic plasmacytoid DC tumor (BPDCN) leukemia, non- It is selected from Hodgkin lymphoma (NHL), mantle cell lymphoma, and Hodgkin leukemia (HL). In another embodiment, the cancer is acute myelogenous leukemia (AML). In yet another embodiment, the acute myelogenous leukemia is refractory or recurrent acute myelogenous leukemia. In another embodiment, the invention provides treatment of a patient with multidrug resistant AML. P-glycoprotein (PGP), also known as MDR1, is a 170 kD ATP-dependent drug efflux pump. It is a member of the ABC superfamily, is abundantly expressed in multidrug resistant (MDR) cells and is made by the ABCB1 gene. AML cells expressing PGP are, at least in part, resistant to treatment with conventional chemotherapy. Accordingly, the present invention also provides a method of treating AML expressing PGP.

The present invention provides a hematological cancer having at least one negative prognostic factor, such as overexpression of P-glycoprotein, overexpression of EVI1, p53 mutation, DNMT3A mutation, FLT3 internal tandem duplication, and/or complex karyotype. A method of treatment is also provided. In another embodiment, the invention also provides a method of treating a hematological cancer having reduced expression of BRCA1, BRCA2, or PALB2 or a mutation of BRCA1, BRCA2, or PALB2. Also within the scope of the present invention is BRCA1, BRCA2, prior to administration of at least one negative prognostic factor (eg at least one poor cytogenetic factor), and/or anti-CD33 immunoconjugate (eg IMGN779). Or selection of patients with reduced expression or mutations of PALB2.

The invention also provides a method of treating a blood cancer having a RAS mutation. The invention also provides a method of treating a hematological cancer having a TP53 mutation. The invention also provides a method of treating a blood cancer having an IDH mutation. The invention also provides a method of treating a hematological cancer having an FLT3 mutation.

Single nucleotide polymorphisms (SNPs) are associated with the effectiveness of CD33-directed therapy. For example, patients with the rs12459419T polymorphism (described in, for example, Malik et al., Hum. Mol. Genet. 15:3557-70 (2015)) are less responsive to CD33-directed therapy. Patients with cytosine (C) instead of thymine (T) at the SNP position respond favorably to CD33-directed therapy. Thus, in some embodiments, the present invention provides methods for treating cancer in a patient homozygous for rs12459419C(CC). In some embodiments, the invention provides methods of treating cancer in a patient heterozygous for rs12459419C (eg, CT). Patients undergoing CD33-directed therapy with the rs351112940A polymorphism (as described in, eg, Mortland et al., Clin. Cancer Res. 19: 1620-27 (2013)) have a major allele, rs35112940G. Poor overall survival and increased risk of recurrence compared to homozygous patients. Thus, in some embodiments, the present invention provides methods for treating cancer in a patient homozygous for rs351112940G (GG). In some embodiments, the invention provides methods for treating cancer in a patient heterozygous for rs351112940G (eg, GA). Patients undergoing CD33-directed therapy with the rs1803254C polymorphism (as described in, for example, Mortland et al.) have a higher 3-year treatment association compared to patients homozygous for the major allele rs1803254G. Indicates the mortality rate. Thus, in some embodiments, the present invention provides methods of treating cancer in a patient homozygous for rs1803254G(GG). In some embodiments, the invention provides methods for treating cancer in a patient heterozygous for rs1803254G (eg, GC).

In some embodiments, the method comprises administering an anti-CD33 immunoconjugate (eg, IMGN779) to the patient and the homozygous rs12459419C genotype has been detected in a sample obtained from the patient. In some embodiments, the method comprises detecting a homozygous rs12459419C genotype in a sample obtained from the patient and then administering an anti-CD33 immunoconjugate (eg, IMGN779).

In some embodiments, the CD33 SNP is detected in a blood sample obtained from the patient. In some embodiments, the CD33 SNP is detected in a buccal swab obtained from the patient.

In some embodiments, the cancer is a CD33-positive cancer. In some embodiments, at least 20% of cancer-derived myeloblasts (“blasts”) are CD33-positive, as measured, for example, by flow cytometry. In some embodiments, cancer-derived blasts are obtained by bone marrow biopsy.

V. Methods of Administering Anti-CD33 Immunoconjugates As provided herein, anti-CD33 immunoconjugates (eg, IMGN779) can be administered at specific doses. Unless otherwise specified (eg, in the examples) mg/kg doses are antibody-based. For example, in some embodiments, about 0.02 to about 0.75 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.3 to about 0.75 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.3 to about 0.7 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.4 to about 0.7 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.5 to about 0.7 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.39 to about 0.75 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.39 to about 0.7 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered.

In some embodiments, about 0.3 to about 0.8 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.4 to about 0.8 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.5 to about 0.8 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered.

In some embodiments, about 0.3 to about 0.9 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.4 to about 0.9 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.5 to about 0.9 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered.

In some embodiments, about 0.3 to about 1.0 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.4 to about 1.0 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.5 to about 1.0 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered.

In some embodiments, about 0.3 to about 1.2 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.4 to about 1.2 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.5 to about 1.2 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered.

In some embodiments, about 0.3 to about 1.25 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.4 to about 1.25 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.5 to about 1.25 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered.

In some embodiments, about 0.54 to about 0.75 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly. In some embodiments, about 0.54 to about 0.7 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly. In some embodiments, about 0.54 to about 0.8 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly. In some embodiments, about 0.54 to about 0.9 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly. In some embodiments, about 0.54 to about 1.0 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly. In some embodiments, about 0.7 to about 1.1 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly. In some embodiments, about 0.7 to about 1.15 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly. In some embodiments, about 0.7 to about 1.2 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly. In some embodiments, about 0.7 to about 1.25 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly.

In some embodiments, about 1 to about 1.6 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly. In some embodiments, about 1 to about 2 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly.

In some embodiments, about 1.2 to about 1.6 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly. In some embodiments, about 1.2 to about 2 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly.

In some embodiments, about 1.5 to about 1.6 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly. In some embodiments, about 1.5 to about 2 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, biweekly.

In some embodiments, about 0.3 to about 0.54 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, once weekly. In some embodiments, about 0.39 to about 0.54 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, once weekly. In some embodiments, about 0.54 to about 0.7 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered, eg, once weekly.

In some embodiments, about 0.39 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.54 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.55, 0.56, 0.57, 0.58, 0.59, 0.60, 0.61, 0.62, 0.63, 0.64, 0. 65, 0.66, 0.67, 0.68, or 0.69 mg/kg of anti-CD33 immunoconjugate (eg IMGN779) is administered. In some embodiments, about 0.70 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered. In some embodiments, about 0.71, 0.72, 0.73, 0.74, 0.75, 0.76, 0.77, 0.78, 0.79, or 0.80 mg/kg. Of the anti-CD33 immunoconjugate (eg, IMGN779).

In some embodiments, about 0.54 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.55, 0.56, 0.57, 0.58, or 0.59 mg/kg anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, for example. , On days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.6 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.61, 0.62, 0.63, 0.64, 0.65, 0.66, 0.67, 0.68, or 0.69 mg/kg of anti-CD33 immunogen. The conjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.7 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.75 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.8 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.85 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.90 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.91 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.92 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.93 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.94 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.95 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.96 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.97 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.98 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.99 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 1 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 1.01 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.02 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.03 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.04 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.05 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.06 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.07 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.08 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.09 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 1.1 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 1.11 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.12 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.13 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.14 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.15 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.16 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.17 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.18 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.19 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 1.2 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 1.21 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.22 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.23 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.24 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.25 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 1.5 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.51 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.52 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.53 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.54 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.55 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.56 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.57 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.58 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.59 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle. In some embodiments, about 1.6 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 1.7 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 1.8 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 1.9 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 1.95 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 2 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about every 2 weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, about 0.3 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.35 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.39 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.4 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.45 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.50 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.55 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.6 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.65 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.7 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.75 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.8 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.85 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.90 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.91 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.92 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.93 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.94 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.95 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.96 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.97 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.98 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 0.99 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 1 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle.

In some embodiments, about 1.01 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.02 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.03 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.04 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.05 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.06 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.07 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.08 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.09 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 1.1 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 1.11 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.12 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.13 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.14 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.15 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.16 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.17 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.18 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.19 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 1.2 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 1.21 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.22 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.23 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.24 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.25 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 1.5 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.51 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.52 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.53 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.54 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.55 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.56 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.57 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.58 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.59 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. .. In some embodiments, about 1.6 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 1.7 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 1.8 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 1.9 mg/kg of the anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 1.95 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once a week, eg, on days 1, 8, 15, and 22 of a 28-day cycle. ..

In some embodiments, about 2 mg/kg of anti-CD33 immunoconjugate (eg, IMGN779) is administered about once weekly, eg, on days 1, 8, 15, and 22 of a 28-day cycle.

In some embodiments, the anti-CD33 immunoconjugate (eg, IMGN779) is administered once weekly, eg, on days 1, 8, 15, and 22 of the 28-day cycle.

In some embodiments, the anti-CD33 immunoconjugate (eg, IMGN779) is administered about every two weeks, eg, on days 1 and 15 of a 28-day cycle.

In some embodiments, an anti-CD33 immunoconjugate (eg, IMGN779) is administered to maintain exposure levels or avoid a significant reduction in exposure.

In some embodiments, administration of an anti-CD33 immunoconjugate (eg, IMGN779) results in saturation of residual free CD33.

In some embodiments, administration of administration of an anti-CD33 immunoconjugate (eg, IMGN779) results in a reduction in peripheral hematoblasts. In some embodiments, administration of an anti-CD33 immunoconjugate (eg, IMGN779) results in a reduction in peripheral hemoblasts within 3-8 days of initial administration. In some embodiments, administration of the anti-CD33 immunoconjugate (eg, IMGN779) results in a reduction in peripheral hemoblasts after the second administration. In some embodiments, administration of the anti-CD33 immunoconjugate (eg, IMGN779) is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85% after the second dose. , 90%, 95%, or 100% reduction in peripheral blasts. In some embodiments, administration of an anti-CD33 immunoconjugate (eg, IMGN779) achieves a peripheral hemoblast percentage of less than about 5%, 4%, 3%, 2%, 1%, or 1%. ..

In some embodiments, administration of an anti-CD33 immunoconjugate (eg, IMGN779) results in a reduction in myeloblasts. In some embodiments, administration of an anti-CD33 immunoconjugate (eg, IMGN779) results in a myeloblast reduction of at least 40%. In some embodiments, administration of an anti-CD33 immunoconjugate (eg, IMGN779) results in at least 45% reduction in myeloblasts. In some embodiments, administration of an anti-CD33 immunoconjugate (eg, IMGN779) results in a myeloblast reduction of at least 48%. In some embodiments, administration of the anti-CD33 immunoconjugate (eg, IMGN779) is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95. % Or 100% reduction in myeloblasts. In some embodiments, administration of an anti-CD33 immunoconjugate (eg, IMGN779) results in at least 59% reduction in myeloblasts. In some embodiments, administration of an anti-CD33 immunoconjugate (eg, IMGN779) results in a myeloblast reduction of at least 96%. In some embodiments, administration of an anti-CD33 immunoconjugate (eg, IMGN779) achieves a myeloblast percentage of less than about 5%, 4%, 3%, 2%, 1%, or 1%.

In some embodiments, the anti-CD33 immunoconjugate (eg, IMGN779) is administered to the subject in a pharmaceutically acceptable dosage form. The anti-CD33 immunoconjugate (eg, IMGN779) can be administered intravenously as a bolus or by continuous infusion over a period of time, intramuscular, subcutaneous, intraarticular, intrasynovial, intrathecal, oral, topical. It can be administered topically or by the inhalation route. In some embodiments, the anti-CD33 immunoconjugate (eg, IMGN779) is administered intravenously.

The anti-CD33 immunoconjugates (eg IMGN779) and pharmaceutical compositions used in the methods of the present disclosure can be supplied as solutions or lyophilized powders tested for sterility and endotoxin levels. Suitable pharmaceutically acceptable carriers, diluents and excipients are well known and can be determined by those skilled in the art on the basis of the clinical situation.

The following examples are presented to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the assays, screens, and therapeutic methods of the present invention. It is not intended to limit the scope of what is considered their invention.

Example 1. In vitro potency of IMGN779 in AML cell line IMGN779 is a specific targeting antibody drug conjugate (ADC) that binds CD33 with high affinity and is widely expressed on acute myelogenous leukemia (AML) cells. However, it is not expressed in non-hematopoietic tissues. IMGN 779 is disclosed in International Publication Application Nos. WO 2015/179400 and WO 2012/128868, and US Published Application Nos. US 2017/0080102 and US 2015/009987, each of which is incorporated herein by reference in its entirety. Already mentioned in the issue. IMGN779 contains a humanized anti-CD33 antibody (Z4681A) attached to a DNA alkylated payload DGN462 (approximately 3 payloads per antibody) via a disulfide containing linker. The Z4681A antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO:9 and a variable light chain having the amino acid sequence of SEQ ID NO:10. The Z4681A variable heavy chain comprises the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 1-3, and the Z4681A variable light chain comprises the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 4-6. When released in target cells, DGN462 exerts its antitumor effect by alkylated DNA without cross-linking, leading to cell cycle arrest and apoptosis.

The potency of IMGN779 was tested in two AML cell lines, EOL-1 and MV4-11 in an in vitro cytotoxicity assay.

The cytotoxicity assay was performed in triplicate for each data point. Culture medium was supplemented with 100 nM huKTI to block Fc receptors on AML cells. AML cells were added to each well of a flat bottom 96 well plate. Test articles were diluted in complete cell culture medium and the dilutions added to plate cells. Wells containing cells and media, but no test article, and wells containing media only were included in each assay plate. The plates were incubated for 4 days at 37° C. in a 6% CO ₂ humidified incubator. Thereafter, the relative number of viable cells in each well was determined using a Water Soluble Tetrazolium Salt-8 (WST-8) based cell counting kit. WST-8 reagent was added to 10% of the final volume and the plates were incubated at 37°C for an additional 2-8 hours. The absorbance of each well was measured with a plate reader spectrophotometer at dual wavelength mode 450 nm/650 nm, and the absorbance at 650 nm (non-specific light scattering by cells) was subtracted from the absorbance at 450 nm. Viability of cells in each well was calculated by subtracting the mean value of wells containing medium only and dividing by the mean value of wells containing cells and medium but no test article. Viability was plotted against test article concentration in a semi-log plot. IC50 values were estimated from the plotted survival curves.

As shown in FIGS. 1A and 1B, IMGN779 was cytotoxic to both cell lines tested, killing EOL-1 cells with an IC50 of 10 nM and MV4-11 cells with an IC50 of 5 nM (FIG. 1A and 1B solid lines). Moreover, killing was CD33-dependent, as blocking CD33 with unconjugated antibody (dotted lines in FIGS. 1A and 1B) reduced IMGN779 cytotoxicity by at least 10-fold.

Example 2. In Vitro Cytotoxicity and Apoptotic Activity of DNA Cross-Linking vs. DNA Alkylation Payload in AML Cell Lines DNA Cross-linking Payload in IMGN779, DGN484 and DNA Alkylation Payload, DGN462, Efficacy in Three In Vitro Cytotoxicity Assays Strains, EOL-1, HL-60, and OCI-M1 were tested. DGN484 is disclosed as compound 1H in U.S. Publication No. 2016-0108129.

The cytotoxicity assay was performed in triplicate for each data point. Cell lines were cultured as described above. Cells in culture were plated in 96 well plates at 50,000 viable cells/well. Serial dilutions of either DGN462 or DGN484 were added to cells in 96 well plates in a total volume of 200 μL/well. Wells containing medium only served as a negative control, and cells in medium treated with DGN484/DGN462 solvent at concentrations consistent with the highest serial dilution molarity served as positive controls. After 72 hours of culture, 20 μL of WST-8 was added to each well in 200 μL increments. The plate was replaced with an incubator. Two to eight hours after WST-8 exposure, the relative amount of viable cells was assessed (SpectraMax M2; at 450 nm) using spectrophotometry.

As shown in FIG. 2A, EOL-1 cells are more sensitive to DGN462 and DGN484 than HL60 and OCI-M1 cells. Furthermore, the crosslinker, DGN484, is more potent in vitro than the alkylating agent, DGN462, in all cell lines tested.

The apoptotic activity of crosslinkers and alkylating agents was also tested in these AML cell lines. In these experiments, cell lines were cultured as described above. Cells in culture were harvested and seeded at 500,000 viable cells/mL in cell culture flasks. Either DGN462, DGN484, or DGN462/DGN484 vehicle (negative control) was added to the flask. After 24 and 48 hours of culture growth, 2 mL per culture was harvested and stained with fluorochrome conjugated Annexin V and To-Pro®-3 iodide. Cells were then run on a BD FACSCanto™ II and the results were treated with FACSDiva 10 and graphed.

As shown in Figure 2B, in vitro treatment with DGN484 resulted in more apoptotic cells in the cell lines tested than treatment with an equivalent dose of DGN462. The magnitude of this difference was greater in cell lines with greater DGN sensitivity.

Example 3. In Vitro Cytotoxicity of DNA Cross-Linking vs. DNA Alkylation Conjugates in AML Cell Lines IMGN779 Conjugate and CD33 Targeting Cross-Linker Conjugate (ie, Z4681A Conjugate to DNA Cross-Linking Payload, DGN484 via sSPDB Linker) Antibody) was tested on an AML cell line in an in vitro cytotoxicity assay.

The assay was run in triplicate for each data point. Culture medium was supplemented with 100 nM huKTI to block Fc receptors on AML cells. AML cells, 1500-10,000 in 100 μL complete culture medium were added to each well of a flat bottom 96 well plate. Test articles were diluted in complete cell culture medium and 100 μL of diluent was added to plate cells. Wells containing cells and media, but no test article, and wells containing media only were included in each assay plate. The plates were incubated for 4-7 days at 37° C. in a 6% CO ₂ humidified incubator. The relative number of viable cells in each well was then determined using a Water Soluble Tetrazolium Salt-8 (WST-8) based cell counting kit (Dojindo Molecular Technologies, Inc., Rockville, MD). WST-8 reagent was added to 10% of the final volume and the plates were incubated at 37°C for an additional 2-6 hours. The absorbance of each well was measured on a plate reader spectrophotometer at dual wavelength mode 450 nm/650 nm, then the absorbance at 650 nm (non-specific light scattering by cells) was subtracted from the absorbance at 450 nm. Viability of cells in each well was calculated by subtracting the mean value of wells containing medium only and dividing by the mean value of wells containing cells and medium but no test article. Viability was plotted against test article concentration in a semi-log plot. IC ₅₀ values were estimated from the plotted survival curves.

Binding of IMGN779 and CD33 targeting crosslinker conjugates to MOLM-13 cells is similar to that assessed by indirect binding assay (see Figure 3). As shown in Figures 4A-4D, the CD33 targeting conjugate of DGN484 is 1.6-5 fold more active in vitro than IMGN779 against the AML cell line.

Example 4. Tolerability of IMGN779, Z4681A-s-SPDB-DGN484, and related conjugates in mice, rats, and cynomolgus monkeys. IMGN779 Single-dose Tolerance Intravenous in Female CD-1 Mice To test the tolerability of IMGN779 in vivo, a mouse model was used as described in the protocol below.

Female CD-1 mice are vehicle, 650 μg/kg (32.9 mg antibody/kg by DGN462, 32.9 mg antibody/kg by DGN462) IMGN779, 725 μg/kg (36.7 mg antibody/kg by DGN462, by IMGN779, 800 μg/kg (DGN462). 40.5 mg antibody/kg) IMGN779, 875 μg/kg (DGN462, 44.3 mg antibody/kg) IMGN779, 950 μg/kg (DGN462, 48.1 mg antibody/kg) IMGN779, or 1025 μg/kg (44.3 mg antibody/kg). A single intravenous injection of 51.9 mg antibody/kg of IMGN779 with DGN462 was received in the lateral tail vein. The antibody does not cross-react with mouse CD33, making this in vivo mouse model an indicator of off-target toxicity only. Mice were observed daily for 19 days to determine body weight. Animals were euthanized if they experienced more than 20% weight loss or were moribund. The BW change rate is calculated from the following formula:% BW change=[(after BW/before BW)-1]×100; after BW is the weight after treatment, and before BW is the starting weight before treatment. Is. A positive %BW change indicates BW increase, while a negative %BW change indicates BW loss.

The results are summarized in Table 3 (below) and Figure 5. IMGN779 was tolerated at 650 μg/kg (according to DGN462, 32.9 mg antibody/kg). The nadir of the mean change in body weight was a 4% decrease, occurring on day 3. None of the 8 mice in this treatment group were euthanized due to weight loss. IMGN779 was tolerated at 725 μg/kg (according to DGN462, 36.7 mg antibody/kg). The nadir of the mean change in body weight was a 6% decrease, occurring on day 5. None of the 8 mice in this treatment group were euthanized due to weight loss. IMGN779 was tolerated at 800 μg/kg (40.5 mg antibody/kg according to DGN462). The nadir of the mean change in body weight was an 8% decrease, occurring on day 4. None of the 8 mice in this treatment group were euthanized due to weight loss. IMGN779 was tolerated at 875 μg/kg (according to DGN462, 44.3 mg antibody/kg). The nadir of the average change in body weight was a 6% decrease, occurring on day 6. None of the 8 mice in this treatment group were euthanized due to weight loss. IMGN779 was tolerated at 950 μg/kg (with DGN462, 48.1 mg antibody/kg). The nadir of the mean change in body weight was a 9% decrease, occurring on day 5. None of the 8 mice in this treatment group were euthanized due to weight loss. This dose defines the maximum tolerated dose (MTD) of IMGN779 in mice. IMGN779 was not tolerated at 1025 μg/kg (according to DGN462, 51.9 mg antibody/kg). The nadir of the average change in body weight was an 11% decrease, occurring on day 6. Two of the eight mice in this treatment group were euthanized by weight loss.

B. Tolerability of a single intravenous dose of IMGN779 or Z4681A-s-SPDB-DGN484 in female CD-1 mice. To test the in vivo tolerability of IMGN779 or Z4681A-s-SPDB-DGN484, the mouse model was Used as described in the protocol.

Female CD-1 mice were vehicle 475 μg/kg (24.7 mg antibody/kg by DGN462) IMGN779, 475 μg/kg (25.7 mg antibody/kg by DGN484) Z4681A-s-SPDB-DGN484, 360 μg/kg (19.4 mg antibody/kg by DGN484) Z4681A-s-SPDB-DGN484, 250 μg/kg (13.5 mg antibody/kg by DGN484) Z4681A-s-SPDB-DGN484, or 190 μg/kg. A single intravenous injection of Z4681A-s-SPDB-DGN484 (10.3 mg antibody/kg by DGN484) was received in the lateral tail vein. The antibody does not cross-react with mouse CD33, making this in vivo mouse model an indicator of off-target toxicity only. Mice were observed daily for 90 days to determine body weight. Animals were euthanized if they experienced more than 20% weight loss or were moribund. The BW change rate is calculated from the following formula:% BW change=[(after BW/before BW)-1]×100; after BW is the weight after treatment, and before BW is the starting weight before treatment. Is. A positive %BW change indicates BW increase, while a negative %BW change indicates BW loss.

The results are summarized in Table 4 (below) and Figure 6. IMGN779 was tolerated at a dose of 475 μg/kg (with DGN462, 24.7 mg antibody/kg) and no animals were euthanized before the end of the study (day 90). No other clinical observations were made during the study, except for minimal individual animal BW loss. The group average (N=8) BW loss nadir occurred on day 4 (-4.2%).

Z4681A-s-SPDB-DGN484 at a dose of 190 μg/kg (10.3 mg antibody/kg according to DGN484) was unacceptable, and before the end of the study, 4 of 8 animals had more than 20%. BW loss of 1 animal (one on days 24, 33, 47 and 57), 2 animals were euthanized with >20% BW loss with hind limb paralysis. (Both animals on day 33). Two of the eight mice survived until the end of the study (day 90). The group average (N=8) BW loss nadir occurred on day 24 (-5.2%). Z4681A-s-SPDB-DGN484 at a dose of 250 μg/kg (13.5 mg antibody/kg according to DGN484) was unacceptable and, prior to the end of the study, 2 of 8 animals had >20%. BW loss (one on days 20 and 23) and 3 animals euthanized by hindlimb paralysis (one on days 28, 29 and 30). Two animals were euthanized by weight loss with hind limb paralysis (day 26, 28) and one animal was euthanized by scoliosis (day 20). The group average (N=8) BW loss nadir occurred on day 20 (-10.4%). Z4681A-s-SPDB-DGN484 at a dose of 360 μg/kg (according to DGN484, 19.4 mg antibody/kg) was unacceptable, and before the end of the study, 5 out of 8 mice had >20%. Euthanized by BW loss (2 at day 17 and 1 at day 23, 24 and 26), 3 mice were euthanized by >20% BW loss with hindlimb paralysis. They were killed (one each on day 16, 19, and 28). The group average (N=8) BW loss nadir occurred on day 16 (-13.5%). Z4681A-s-SPDB-DGN484 at a dose of 475 μg/kg (25.7 mg antibody/kg by DGN484) was unacceptable, and before the end of the study, 4 of 8 animals had >20%. Were euthanized by BW loss of 1 (one each on days 17, 18, 23 and 26) and 3 animals were euthanized due to morbidity (day 13, 19). And 1 animal each on day 20), one animal was euthanized due to morbidity with hind limb paralysis (day 18). Group mean (N=8) BW loss nadir occurred on day 4 (-5.3).

A serious clinical observation noted in mice treated with each of four different dose levels of Z4781A-s-SPDB-DGN484 (190, 250, 360 or 475 μg/kg, based on DGN484 concentration) was hindlimb paralysis. Yes, suggesting that treatment with this conjugate ultimately has some neurological effects in these non-tumor bearing CD-1 mice. Hindlimb paralysis was not observed in any of the 8 mice treated with 475 μg/kg of IMGN779 (based on DGN462 concentration).

C. Toxicity of IMGN779 in mice, rats, and cynomolgus monkeys (in a contract research institution) In three different toxicology studies performed in three different species, IMGN779 was administered as a single intravenous injection to animals at 36 mg/kg (antibody CD-1 mice at 5 mg/kg (by antibody) and female Sprague Dawley rats at 2.4 mg/kg (by antibody) and cynomolgus monkeys at 2.4 mg/kg (by antibody). Liver enzymes and blood cellularity were measured by standard methods. Body weight was measured and clinical observations recorded over the duration of each study. At the end of each study, animals were euthanized, organs and tissues were collected, fixed, processed on hematoxylin and eosin (H&E) stained slides, examined by a veterinary pathologist and their findings recorded. The results of these three toxicology tests are summarized in Table 5 (below).

In mice, IMGN779 given as a single intravenous injection at 36 mg/kg (by antibody) caused a mild elevation of liver enzymes (ALT, AST, SDH). The major histopathological findings of interest were gastrointestinal (GI) disorders and bone marrow depletion. The primary clinical observation was weight loss. When analyzed for peripheral blood cellularity, a decrease was seen in platelets (mild to moderate), reticulocytes (severe), neutrophils (moderate), and lymphocytes (moderate).

In rats, IMGN779 given as a single intravenous injection at 5 mg/kg (by antibody) caused a slight increase in SDH and a slight decrease in ALT and ALP. The major histopathological findings of interest were gastrointestinal (GI) disorders and bone marrow depletion. The main clinical observations were weight loss and diarrhea at doses above the maximum tolerated dose (MTD). When analyzed for peripheral blood cellularity, a decrease was seen in platelets (mild to moderate), reticulocytes (severe), neutrophils (moderate to severe), and lymphocytes (moderate).

In cynomolgus monkeys, IMGN779 administered as a single intravenous injection at 2.4 mg/kg (by antibody) did not cause changes in the levels of liver enzymes (ALT, AST, SDH). The major histopathological findings of interest were gastrointestinal (GI) disorders and bone marrow depletion. The main clinical observations were weight loss and diarrhea. When analyzed for peripheral blood cellularity, reductions were found in platelets (mild to moderate), reticulocytes (severe), neutrophils (moderate to severe), and lymphocytes (mild).

D. Toxicity of CD33 targeting conjugates without IMGN779 (cleavable linker) and cleavable linker (Z4681A-IGN23) in female CD-1 mice Cleavable -s-SPDB-linker or Z4681A-IGN23. Immun 779 with a monoimine payload containing conjugate with a non-cleavable linker was administered to female CD-1 mice as a single intravenous injection. IGN23 is disclosed in US Publication No. 2016-0222013 as compound 254C. IMGN779 is administered at a dose of approximately 12.5 mg/kg (by antibody) and approximately 35 mg/kg (by antibody) and Z4681A-IGN23 is administered at a dose of approximately 10 mg/kg (by antibody). Blood samples were collected 5 days after administration of the conjugate and analyzed for liver enzyme (ALT, AST, SDH) levels and cellularity (white blood cells, platelets, reticulocytes, neutrophils and lymphocytes) using standard methods. did. The percent change (% change) for each parameter was calculated as follows:% change = ((value, conjugate treated mouse-value, untreated mouse)/value, untreated mouse) x 100%, normal The historical value of treated female CD-1 mice was used as the untreated baseline level for each parameter. A positive% change represents an increase compared to the baseline level, a negative% change represents a decrease compared to the baseline level, and a 100% decrease indicates that the value of the parameter has dropped to zero. The results are summarized in Table 6 (below).

Treatment with a low dose of 12.5 mg/kg IMGN779, 5 days after conjugate administration, resulted in a mild increase in liver enzymes, a 45% increase in ALT, a 39% increase in AST, and a 46% increase in SDH. .. Similarly, treatment with a 35 mg/kg dose of IMGN779 increased liver enzymes, but not more than the 12.5 mg/kg dose, with a 393% increase in ALT, a 135% increase in AST, and a 275% increase in SDH. It is an increase. In contrast, treatment with the 10 mg/kg dose of Z4681A-IGN23 resulted in a very large increase in liver enzymes, above the baseline level, a 14,000% increase in ALT, a 6,000% increase in AST, This is a 750% increase in SDH.

Treatment with low doses of 12.5 mg/kg IMGN779 5 days after conjugate administration resulted in leukocytes (WBC, 67%), reticulocytes (98%), neutrophils (83%) and lymphocytes (63%). And a very small increase in platelets (9.4%). In contrast, treatment with Z4681A-IGN23 at a dose of 10 mg/kg (by antibody) (equivalent to 80% of low dose IMGN779) resulted in a 91% reduction in platelets. In addition, treatment with a 10 mg/kg dose of Z4681A-IGN23 resulted in a 10% reduction in WBC, a 73% reduction in reticulocytes, a 200% increase in neutrophils and a 47% decrease in lymphocytes. Treatment with high dose IMGN779, 35 mg antibody/kg results in a 79% reduction in WBC, a 77% reduction in platelets, a 99% reduction in reticulocytes, a 92% reduction in neutrophils, and a 76% reduction in lymphocytes.

Example 5. In vivo efficacy of a single dose of IMGN779 in the EOL-1 subcutaneous model. Data collection and analysis of all subcutaneous xenograft models.
Mice were weighed twice weekly and monitored for clinical signs during the study period. Animals were euthanized when hind limb paralysis appeared, body weight was lost by more than 20% of pre-treatment weight, tumor ulceration occurred, or any signs of distress were visible. Tumor volume was measured 1-2 times weekly in three dimensions using calipers. Tumor volume was expressed in mm ³ using the formula V=length×width×height×1/2 (Tomayko and Reynolds, Cancer Chemother. Pharmacol. 24:148-54 (1989)). Activity is as described by Bissey et al. , Cancer Res. 51: 4845-52 (1991). Tumor growth inhibition (T/C value) was evaluated using the following formula: T/C (%)=(median tumor volume of treatment/median tumor volume of control)×100%. Tumor volume was determined simultaneously for the treated (T) and vehicle control (C) groups when the vehicle control tumor volume reached a given size (Bissery et al., Cancer Res. 51:4845-52(. 1991.) The median daily tumor volume for each treatment group, including tumor-free mice (0 mm ³ ), was determined.By NCI standard, T/C<42% is the minimum level of antitumor activity. T/C<10% is considered a high level of antitumor activity.

To test the efficacy of IMGN779 for its ability to reduce tumor burden in vivo, a subcutaneous tumor model was used as described in the protocol below.

Each of the female athymic nude mice was inoculated subcutaneously on the right flank with 1×10 ⁷ EOL-1 cells, human AML cell line in 100 μl of serum-free medium/matrigel. Six days after EOL-1 inoculation, mice were randomized into test groups based on tumor volume. Twenty-four hours prior to conjugate administration, mice were injected intraperitoneally with 400 mg/kg of non-targeted humanized AVE1642 IgG antibody, blocking Fc receptors on EOL-1 cells and conjugating nonspecific conjugates. Prevented capture. Similarly, 5 and 10 days after conjugate administration (10 and 15 days after EOL-1 inoculation), mice were injected ip with 100 mg/kg AVE1642 antibody to detect Fc receptors on EOL-1 cells. Ensured continuous interruption. Six days after EOL-1 inoculation, mice received vehicle, 10 μg/kg (by DGN462; 0.602 mg antibody/kg) IMGN779, 30 μg/kg (by DGN462; 1.80 mg antibody/kg) IMGN779, 60 μg/kg. (By DGN462; 3.61 mg antibody/kg) IMGN779, 10 μg/kg (by DGN462; 0.424 mg antibody/kg) chKTI-DGN462, 30 μg/kg (by DGN462; 1.27 mg antibody/kg) chKTI- A single intravenous injection of DGN462 or chKTI-DGN462 at 60 μg/kg (according to DGN462; 2.54 mg antibody/kg) was received in the lateral tail vein.

The results are shown in Table 7 (below) and FIG. 7. IMGN779 was highly active at all dose levels tested in this study. IMGN779 at both 60 and 30 μg/kg had T/C values of 0% and 6/6 complete regression (CR) and all animals were tumor free until the end of the study on day 90. However, one animal needed to be sacrificed in the highest dose group at day 67 due to morbidity. At the lowest dose of 10 μg/kg, IMGN779 had a T/C value of 3% with 6/6 being partial regression (PR) and 6/6 being CR. The non-targeted chKTI-DGN462 conjugate is inactive at 10 μg/kg dose, 78% T/C, no PR or CR, active at 30 μg/kg, T/C 20%. , 1/6 was PR and only 1/6 was CR. At the highest dose of 60 μg/kg, the chKTI-DGN462 non-targeting conjugate was highly active with T/C values of 3%, 5/6 PR and 5/6 CR.

Example 6. In Vivo Efficacy of a Single Dose of IMGN779 in the HL60/QC Subcutaneous Model To test the efficacy of IMGN779 for its ability to reduce tumor burden in vivo, a subcutaneous tumor model was used as described in the protocol below.

Female C. Each of the B17 SCID mice was inoculated subcutaneously on the right flank with 5×10 ⁶ HL60/QC cells, human AML cell line in 100 μl serum-free medium/matrigel. 9 days after HL60/QC inoculation, mice were randomized into test groups. Twenty-four hours prior to conjugate administration, mice were injected ip with 400 mg/kg of non-targeted chKTI antibody to block Fc receptors on HL60/QC cells and prevent non-specific uptake of conjugate. did. Similarly, 4 and 9 days post conjugate administration, mice were injected ip with 100 mg/kg of non-targeted chKTI Ab to ensure continued blockade of Fc receptors on HL60/QC cells. 10 days after HL60/QC inoculation, the mice received vehicle, 10 μg/kg (with DGN462; 0.520 mg antibody/kg) IMGN779, 5 μg/kg (with DGN462; 0.260 mg antibody/kg) IMGN779 or 2.5 μg. A single intravenous injection of IMGN779/kg (according to DGN462; 0.130 mg antibody/kg) was received in the lateral tail vein.

The results are shown in Table 8 (below) and FIG. IMGN779 at a dose of 10 μg/kg (according to DGN462) was highly active, resulting in T/C of 7% and complete regression (CR) of 1/6. IMGN779 at both the 5 μg/kg and 2 μg/kg doses (according to DGN462) were inactive, with 46% T/C (0/6 CR) and 76% T/C (0/6 CR, respectively). ) Was brought. These results define the minimum effective dose of a single intravenous dose of IMGN779 in the HL60/QC subcutaneous model as 10 μg/kg.

Example 7. In Vivo Efficacy of Multiple Doses of IMGN779 in HL60/QC Subcutaneous Model To test the efficacy of IMGN779 for its ability to reduce tumor burden in vivo, a subcutaneous tumor model was used as described in the protocol below.

Female C. Each of the B17 SCID mice was inoculated subcutaneously on the right flank with 5×10 ⁶ HL60/QC cells, human AML cell line in 100 μl serum-free medium/matrigel. Unlike previous studies, FcR was shown to not contribute to activity at the low doses tested in this study, so the group was treated with excess human IgG blocking FcR to eliminate nonspecific activity. I didn't. 9 days after cell inoculation, mice were randomized into test groups based on tumor volume. Nine days after HL60/QC inoculation (Day 1 of treatment), mice received vehicle, 10 μg/kg (with DGN462; 0.506 mg antibody/kg) IMGN779, 30 μg/kg (with DGN462; 1.52 mg antibody/kg). ) IMGN779, or 0.66 mg/kg unconjugated Z4681A antibody + 30 μg/kg free DGN462 payload, received a single intravenous injection in the tail vein. One particular group in this study was on days 4 and 7 (Q3D x 3 dose completion) or 8 and 15 (QW x 3 dose completion) after the first IMGN779 treatment (day 1). Either received an additional single intravenous injection of IMGN779 at 10 μg/kg (by DGN462; 0.506 mg antibody/kg) into the tail vein. Certain other groups in this study were on Days 4 and 7 (Completion of Q3D x 3 Dosing Scheme) or Days 8 and 15 (Completion of QW x 3 Dosing Schedule) after the first IMGN779 treatment (Day 1). ), received an additional single intravenous injection of IMGN779 at 30 μg/kg (by DGN462; 1.52 mg antibody/kg) into the tail vein.

The results are shown in Table 9 (below) and FIG. 9. IMGN779 was active at 10 μg/kg (according to DGN462) at both single dose and QW×3 doses (days 1, 8 and 15) with T/C values of 12% and 10%, respectively. It was IMGN779 was highly active when administered at 10 μg/kg (according to DGN462) Q3D×3 (Days 1, 4 and 7) with a T/C value of 7%. In the single dose group, 3/6 had partial regression (PR), 3/6 had complete regression (CR), and no animals remained tumor free at the end of the study (day 90). In both the QW×3 and Q3D×3 groups, 6/6 were PR, 6/6 were CR, and 2/6 of the animals in each group were tumor free at the end of the study (day 90). At the high dose of 30 μg/kg (according to DGN462), IMGN779 was highly active on all dosing schedules, with a single dose of 7% T/C value, 6% on Q3D×3 and QW×3 groups. T/C value of All three groups had 6/6 PR and 6/6 CR. At the end of the study (Day 90), the single dose group and the QW×3 group had 5/6 tumor free survivors and the Q3D×3 group had 4/6 tumor free survivors. A single dose of Z4681A antibody at 0.66 mg/kg and a single dose of free DGN462 payload at 30 μg/kg was inactive at 92% T/C and 0/6 PR and CR.

Example 8. In Vivo Efficacy of a Single Dose of IMGN779 or Z4681A-s-SPDB-DGN484 in the HL60/QC Subcutaneous Model To test the efficacy of IMGN779 for its ability to reduce tumor burden in vivo, the subcutaneous tumor model follows the protocol below. Used as described.

Female C. Each of the B17 SCID mice was inoculated subcutaneously on the right flank with 5×10 ⁶ HL60/QC cells, human AML cell line in 100 μl serum-free medium/matrigel. 9 days after HL60/QC inoculation, mice were randomized into test groups. Twenty-four hours prior to conjugate administration, mice were injected ip with 400 mg/kg of non-targeting chKTI antibody to block Fc receptors on HL60/QC cells and prevent non-specific uptake of conjugate. .. Similarly, 4 and 9 days post conjugate administration, mice were injected ip with 100 mg/kg of non-targeted chKTI Ab to ensure continued blockade of Fc receptors on HL60/QC cells. 10 days after HL60/QC inoculation, mice received vehicle, 30 μg/kg (with DGN462; 1.56 mg antibody/kg) IMGN779, 10 μg/kg (with DGN462; 0.522 mg antibody/kg) IMGN779, 10 μg/kg. (By DGN484; 0.473 mg antibody/kg) chKTI-s-SPDB-DGN484, 10 μg/kg (0.541 mg antibody/kg by DGN484) Z4681A-s-SPDB-DGN484, 5 μg/kg (by DGN484, 0) .271 mg antibody/kg) Z4681A-s-SPDB-DGN484, 2.5 μg/kg (with DGN484, 0.135 mg antibody/kg) Z4681A-s-SPDB-DGN484, 1 μg/kg (with DGN484, 0.0541 mg). Antibody/kg) Z4681A-s-SPDB-DGN484 or 0.5 μg/kg (0.0271 mg antibody/kg according to DGN484) Z4681A-s-SPDB-DGN484 received a single intravenous injection in the tail vein.

The results are shown in Table 10 (below) and FIG. IMGN779 at a dose of 30 μg/kg (according to DGN462) was highly active, resulting in 4% T/C, 6/6 PR, and 6/6 CR. IMGN779 at a dose of 10 μg/kg was active, resulting in 15% T/C, 4/6 PR, and 3/6 CR. The 10 μg/kg dose (by DGN484) of Z4681A-s-SPDB-DGN484 was highly active, resulting in 5% T/C, 6/6 PR, and 4/6 CR. The doses of 5, 2.5, 1, and 0.5 μg/kg of Z4681A-s-SPDB-DGN484 were all inactive, yielding% T/C values of 43, 88, 97, and 99, respectively. PR or CR was not achieved in any of the treatment groups. The non-targeted chKTI-s-SPDB-DGN484 control conjugate administered at 10 μg/kg (by DGN484) was inactive, yielding a T/C value of 104% and no PR or CR.

Example 9. In Vivo Efficacy of Single vs. Multiple Dose Administration of IMGN779 in MV4-11 Subcutaneous Model To test the efficacy of IMGN779 for its ability to reduce tumor burden in vivo, a subcutaneous tumor model is described in the protocol below. Used as.

Female C. Each of the B17 SCID mice was inoculated subcutaneously on the right flank with 10×10 ⁶ MV4-11 cells, human AML cell line in 100 μl serum-free medium/matrigel. Thirteen days after HL60/QC inoculation, mice were randomized into test groups based on tumor volume. Twenty-four hours prior to conjugate administration, mice were injected intraperitoneally with 400 mg/kg of non-targeting chKTI antibody to block Fc receptors on MV4-11 cells and prevent non-specific uptake of conjugate. .. Similarly, 18 and 23 days after MV4-11 inoculation, mice were injected ip with 100 mg/kg of non-targeted chKTI Ab to ensure continued blockade of Fc receptors on MV4-11 cells. .. Fourteen days after MV4-11 inoculation (Day 1 of treatment), mice received vehicle, 10 μg/kg (with DGN462; 0.506 mg antibody/kg) IMGN779 or 5 μg/kg (with DGN462; 0.253 mg antibody/kg). ) IMGN779 as a single intravenous injection into the lateral tail vein. After this initial administration, one animal in the 5 μg/kg IMGN779-administered group received a second single intravenous injection of 5 μg/kg (according to DGN462) IMGN779 on day 4 of treatment (completion of Q3D×2 dosing regimen). Others receiving the 5 μg/kg IMGN779 group received a second single intravenous injection of 5 μg/kg (according to DGN462) of IMGN779 on day 8 of treatment (QW×2 completion of dosing regimen). Received

The results are shown in Table 11 (below) and FIG. 11. IMGN779 is highly active at a single dose of 10 μg/kg (by DGN462) and both split dose schedules (5 μg/kg, Q3D×2 and QW×2), with 0% T/T in all groups. It was a C value. Complete regression in 6/6 animals in the 10 μg/kg single dose group, 5/6 animals in the 5 μg/kg Q3D×2 group, and 6/6 animals in the 5 μg/kg QW×2 split dose group (CR). Each of the two split-dose groups had more tumor-free survivors (in both groups) at the end of the study (day 90) compared to the single-dose group where 3/6 were tumor-free survivors. It was 5/6).

Example 10. In vivo efficacy of a single dose of IMGN779 in the MV4-11 disseminated model. Data collection and analysis of all disseminated xenograft models.
Mice were weighed twice weekly and monitored for clinical signs during the study period. The measurement end point was the survival rate. Animals were euthanized when hind limb paralysis appeared, body weight lost >20% of pre-treatment weight, visible tumors appeared, or any signs of distress appeared. Natural deaths were recorded if found.

For the disseminated model, tumor growth delay was calculated as T-C, where T is the median survival (days) of the treatment group and C is the median survival (days) of the vehicle control group. .. The rate of life extension of the disseminated model (% ILS) was calculated using the following formula:% ILS=(TC)/C×100%. Antitumor activity was assessed per NCI standard in the disseminated model: ILS>25% is minimal activity, ILS>40% is active, and ILS>50% is highly active.

To test the efficacy of IMGN779 for its ability to reduce tumor burden in vivo, a disseminated tumor model was used as described in the protocol below.

Female NOD SCID mice were pretreated with 150 mg/kg cyclophosphamide to partially engraft the bone marrow to improve engraftment of MV4-11 cells. Cyclophosphamide (Baxter, Lot #4E011, Deadline 05/2017) was reconstituted with 0.9% NaCl and intraperitoneally injected into mice 3 and 2 days prior to day 0 MV4-11 cell inoculation. Was administered. After cyclophosphamide treatment as described above, each mouse was intravenously infused into the tail vein with 3×10 ⁶ MV4-11 cells, a human AML cell line, in 100 μl of serum-free medium. Six days after MV4-11 inoculation, mice were randomized into test groups based on body weight. Twenty-four hours prior to conjugate administration, mice were injected intraperitoneally with 400 mg/kg of non-targeting chKTI antibody to block Fc receptors on MV4-11 AML cells and to induce non-specific uptake of conjugate. Prevented. Seven days after MV4-11 inoculation, mice received vehicle, 10 μg/kg (with DGN462; 0.528 mg antibody/kg) IMGN779, 60 μg/kg (with DGN462; 3.17 mg antibody/kg) IMGN779, 10 μg/kg. Sided with a single intravenous injection of chKTI-s-SPDB-DGN462 (by DGN462; 0.560 mg antibody/kg) or chKTI-s-SPDB-DGN462 at 60 μg/kg (by DGN462; 3.36 mg antibody/kg). Received in the tail vein.

The results are shown in Table 12 (below) and FIG. IMGN779 was very active in this model at 60 μg/kg, with extended lifespan (ILS) of 145.3% and tumor growth delay (TC) of 78.5 days, 3 at the end of the study. One animal survived. IMGN779 was also highly active at 10 μg/kg, ILS was 90.7% at the end of the study, tumor growth delay was 49 days, 1 animal survived. The non-targeting conjugate, chKTI-s-SPDB-DGN462, was active at 60 μg/kg, and at the end of the study, ILS was 42.6%, tumor growth delay was 23 days, and one animal survived. There wasn't. At 10 μg/kg, chKTI-s-SPDB-DGN462 was inactive, at the end of the study, ILS was 4.6%, tumor growth delay was 2.5 days, and no animals survived. ..

Example 11. In vivo efficacy of multiple doses of IMGN779 (QW x 3) in the MV4-11 disseminated model To test the efficacy of IMGN779 for its ability to reduce tumor burden in vivo, the disseminated tumor model is described in the protocol below. Used as described.

Female NOD SCID mice were pretreated with 150 mg/kg cyclophosphamide to partially engraft the bone marrow to improve engraftment of MV4-11 cells. Cyclophosphamide (Baxter, Lot #4E011, Deadline 05/2017) was reconstituted with 0.9% NaCl and intraperitoneally injected into mice 3 and 2 days prior to day 0 MV4-11 cell inoculation. Was administered. After cyclophosphamide treatment as described above, each mouse was intravenously infused into the tail vein with 3×10 ⁶ MV4-11 cells, a human AML cell line, in 100 μl of serum-free medium. Twenty days after MV4-11 inoculation, mice were randomized into test groups. Twenty-four hours prior to conjugate administration, mice were injected intraperitoneally with 400 mg/kg of non-targeting chKTI antibody to block Fc receptors on MV4-11 AML cells and to induce non-specific uptake of conjugate. Prevented. Similarly, 27 and 34 days after MV4-11 inoculation, mice were injected intraperitoneally with 400 mg/kg of non-targeted chKTI antibody to block Fc receptors on MV4-11 AML cells. 21, 28, and 35 days post MV4-11 inoculation, mice received vehicle, 10 μg/kg (with DGN462; 0.534 mg antibody/kg) IMGN779, 3 μg/kg (with DGN462; 0.160 mg antibody/kg). IMGN779, 1 μg/kg (according to DGN462; 0.0534 mg antibody/kg) IMGN779, 0.3 μg/kg (according to DGN462; 0.0160 mg antibody/kg) IMGN779, 0.1 μg/kg (according to DGN462; 0.00534 mg) Antibody/kg) IMGN779, 0.3 μg/kg (by DGN462; 0.00160 mg antibody/kg) IMGN779, or 10 μg/kg (by DGN462; 0.556 mg antibody/kg) chKTI-s-SPDB-DGN462. A single intravenous injection was received in the lateral tail vein.

The results are summarized in Table 13 (below) and Figure 13. IMGN779 at doses of 10 μg/kg and 3 μg/kg (each administered qw×3) were both highly active, 108% and 73% ILS, respectively, and corresponding tumors at 52 and 35 days, respectively. It caused a growth delay. The 1 μg/kg IMGN779, qw×3 regimen was the reliable minimum effective dose in this study, resulting in 33% ILS and 16 days tumor growth delay. Low dose IMGN 779: 0.3, 0.1 and 0.03 μg/kg, qw×3 at 15%, 29% and 33% ILS, respectively, and tumor at 15, 29 and 33 days respectively. It caused a growth delay. Also, the 10 μg/kg dose of the non-targeted Ab-DGN462 conjugate was highly active, but provided only 61.5% ILS and a tumor growth delay of 29.5 days. This demonstrated the benefit of CD33 targeting of IMGN779 when compared to 108% ILS and 52 days tumor growth delay.

Example 12. In Vivo Efficacy of a Single Dose of IMGN779 or Z6481A-s-SPDB-DGN484 in the MV4-11 Disseminated Model To test the efficacy of IMGN779 for its ability to reduce tumor burden in vivo, the disseminated tumor model was Used as described in protocol.

Female NOD SCID mice were pretreated with 150 mg/kg cyclophosphamide to partially engraft the bone marrow to improve engraftment of MV4-11 cells. Cyclophosphamide (Baxter, Lot #4E011, Deadline 05/2017) was reconstituted with 0.9% NaCl and intraperitoneally injected into mice 3 and 2 days prior to day 0 MV4-11 cell inoculation. Was administered. After cyclophosphamide treatment as described above, each mouse was intravenously infused into the tail vein with 3×10 ⁶ MV4-11 cells, a human AML cell line, in 100 μl of serum-free medium. Seven days after MV4-11 inoculation, mice were randomized into test groups. Twenty-four hours prior to conjugate administration, mice were injected intraperitoneally with 400 mg/kg of non-targeting chKTI antibody to block Fc receptors on MV4-11 AML cells and to induce non-specific uptake of conjugate. Prevented. Similarly, 11 and 16 days after MV4-11 inoculation, mice were injected ip with 100 mg/kg of non-targeting chKTI antibody to block Fc receptors on MV4-11 AML cells. Seven days after MV4-11 inoculation, the mice received vehicle, 2.5 μg/kg (with DGN462; 0.130 mg antibody/kg) IMGN779, 1 μg/kg (with DGN462; 0.0521 mg antibody/kg) IMGN779 or 0. 0.5 μg/kg (by DGN462; 0.0260 mg antibody/kg) IMGN779, 2.5 μg/kg Z4681A-s-SPDB-DGN484 (by DGN484; 0.135 mg antibody/kg), 1 μg/kg (by DGN484; 0.0541 mg antibody/kg) Z4681A-s-SPDB-DGN484, 0.5 μg/kg (by DGN484; 0.0271 mg antibody/kg) Z4681A-s-SPDB-DGN484, 0.25 μg/kg (by DGN484; 0.0135 mg antibody/kg), or 0.1 μg/kg (by DGN484; 0.00541 mg antibody/kg) received a single intravenous injection in the tail vein.

The results are summarized in Table 14 (below) and Figure 14. IMGN779 at 2.5 μg/kg, 1.0 μg/kg, and 0.5 μg/kg doses (according to DGN462) were all highly active, with tumors at 32.5 days, 44 days, and 56.5 days, respectively. It resulted in growth retardation (TC) values and extended life span (ILS) of 64%, 87%, and 112%, respectively. The 2.5 μg/kg, 1 μg/kg, 0.5 μg/kg, 0.25 μg/kg, and 0.1 μg/kg doses (according to DGN484) of Z4681A-s-SPDB-DGN484 are all highly active, Tumor growth delay values of 38.5 days, 46.5 days, 26.5 days, 49 days, and 34.5 days, respectively, and ILS of 76%, 92%, 52%, 97%, and 68%, respectively. Brought.

Example 13. In vivo efficacy of multiple doses of IMGN779 (QWx3) in Molm-13 disseminated model To test the efficacy of IMGN779 for its ability to reduce tumor burden in vivo, a disseminated tumor model is described in the protocol below. Used as described.

Female NOD SCID mice were pretreated with 150 mg/kg cyclophosphamide and partially ablated the bone marrow to improve the engraftment of Molm-13 cells. Cyclophosphamide (Baxter, Lot #4E011, Deadline 05/2017) was reconstituted with 0.9% NaCl and was administered ip to mice 2 days before Molm-13 cell inoculation on day 0. After the cyclophosphamide treatment described above, each mouse was intravenously infused into the tail vein with 2×10 ⁵ Molm-13 cells, a human AML cell line, in 100 μl of serum-free medium. Six days after Molm-13 inoculation, mice were randomized into test groups by body weight. Twenty-four hours prior to administration of conjugate, mice were injected intraperitoneally with 400 mg/kg of non-targeting chKTI antibody to block Fc receptors on Molm-13 AML cells and allow non-specific uptake of conjugate. Prevented. Similarly, 13 and 20 days after Molm-13 inoculation, mice were injected ip with 400 mg/kg of non-targeted chKTI antibody to block Fc receptors on Molm-13 AML cells. At 7, 14 and 21 days post Molm-13 inoculation, mice received vehicle, 10 μg/kg (with DGN462; 0.534 mg antibody/kg) IMGN779, 3 μg/kg (with DGN462; 0.160 mg antibody/kg). IMGN779, 1 μg/kg (according to DGN462; 0.0534 mg antibody/kg) IMGN779, 0.3 μg/kg (according to DGN462; 0.0160 mg antibody/kg) IMGN779, 0.1 μg/kg (according to DGN462; 0.00534 mg) Antibody/kg) IMGN779, 10 μg/kg (by DGN462; 0.556 mg antibody/kg) chKTI-s-SPDB-DGN462, or 3 μg/kg (by DGN462; 0.167 mg antibody/kg) chKTI-s-. A single intravenous injection of SPDB-DGN462 was received in the lateral tail vein.

The results are summarized in Table 15 (below) and Figure 15. IMGN779 at doses of 10, 3, 1 and 0.3 μg/kg (each administered qw×3) were all highly active with >158%, >158%, 126% and 58% ILS respectively. , And >30, >30, 24, and 11 days tumor growth delay, respectively. A 0.1 μg/kg dose of IMGN779 (administered qw×3) was the least effective dose and the least active, resulting in 26% ILS and a 5 day tumor growth delay. In contrast, the 10 μg/kg and 3 μg/kg doses of the non-targeting control conjugate, chKTI-s-SPDB-DGN462 (each administered qw×3) were both inactive, 16% and 16% respectively. Delivering 24% ILS and tumor growth delays of 3 and 4.5 days respectively, demonstrating the CD33 targeting specificity of IMGN779.

Example 14. Phase 1 study of IMGN779 intravenously administered to adult patients with relapsed/refractory CD33-positive acute myeloid leukemia. An open-label, multi-center Phase 1 study was initiated to determine the dose (MTD), dosing schedule, and recommended Phase 2 dose of IMGN779 (RP2D). The study consists of a two-step first-in-human (FIH) (first administered to human) study consisting of 1) a dose escalation step to identify the MTD and dosing schedule; and 2) a dose escalation step. Two dosing schedules: Q2W (Schedule A: IMGN779 administered IV on Days 1 and 15 of 28-day cycle) and QW (Schedule B: IV administered on Days 1, 8, 15, and 22 of 28-day cycle). IMGN 779).

The patient population enrolled in the dose escalation stage is CD33-positive, relapsed or refractory AML (at least 20% of blasts are CD33-positive as determined by flow cytometry performed at a local CLIA accredited laboratory). Adult (at least 18 years of age) patients with Relapsed/refractory AML is defined as 1) primary induction failure after two or more cycles of chemotherapy; 2) first early relapse after less than 6 months remission; and 3) second and subsequent relapses .. The primary purpose of the dose escalation phase is to identify the MTD and recommended schedule for IMGN779.

In some examples, patient blood samples or cheek cell swabs are collected and subjected to polymerase chain reaction (PCR), Sanger sequencing; next generation sequencing (NGS); transcriptase quantitative polymerase chain reaction (RT-qPCR or qPCR). Single nucleotide polymorphism in the CD33 gene (Table 16) using techniques known in the art, such as, but not limited to, or similar techniques for identifying DNA sequences. The type (SNP) was evaluated. Patients may also use CD33 mRNA using techniques known in the art, including, but not limited to, RNA sequencing by quantitative polymerase chain reaction (RT-qPCR or qPCR) or next generation sequencing (NGS). (Full length (FL), Delta-exon 2 (ΔE2) Delta-exon 7a (E7a) and various splice variants of delta-exon 2 and delta-exon 7a (ΔE2-E7a) (Lazlo et al. Oncotarget, Vol. .7, No. 28)) and/or ratios.

The dose escalation phase follows a 3+3 cohort design enrolling 3-6 patients in each cohort. Study enrollment is to administer IMGN779 on Schedule A (Q2W) at a starting dose of 0.02 mg/kg (by huMy9-6), which is 1/6 of HNSTD determined by repeated dose GLP toxicity study in cynomolgus monkeys. Start. Dose escalation for this schedule follows a modified Fibonacci dosing scheme.

A cohort review committee (CRC) will be convened after each cohort to review the safety, PK/Pd, and antitumor activity data collected, and based on these data, CRC will be used to determine the dose at Schedule A. It may be decided to continue the titration, add additional patients at the same dose level, or stop enrollment at that dose level or schedule. The CRC may also recommend starting Weekly Dosing Schedule B. The starting dose for Schedule B is determined by the CRC based on safety, Pd, and PK data such that the maximum of one dose level is below the final dose level cleared by CRC from Schedule A. .. Dose escalation proceeds with Schedule A, as shown in Table 17, until the MTD is defined.

The MTD for a given schedule is determined from an assessment of dose limiting toxicity (DLT) during the first dosing cycle. Meet the definition of dose limitation, but clinically significant toxicities or adverse reactions (TEAEs) that occurred during treatment that occur after Cycle 1 may be considered when determining the MTD on a schedule basis. The use of hydroxyurea or leukocyte depletion during testing is taken into account as it can be a confounding factor.

To better characterize safety, up to 12 patients may receive IMGN779 with an estimated RP2D at each dosing schedule based on CRC and drug sponsor recommendations. The CRC will be recruited to review available safety, PK/Pd, and antitumor activity data and to select the dose and dosing schedule used during the dose escalation part of the study. Given that IMGN779 is the targeted therapy, the available information about the target along with safety, PK, and Pd data will be considered when selecting the recommended dose for the expansion phase.

Patients enrolled in the dose escalation phase are assigned to the cohort as follows: patients with CD33-positive AML with initial relapse (disease that relapses more than 6 months after initial remission). A first-recurrent AML is defined as a disease that relapses more than 6 months after initial remission. A Bayesian continuous reassessment (CRM) approach is used to guide DLT monitoring during the dose escalation phase of the study. Once the putative RP2D and schedule have been selected, the dose escalation phase of the study begins. The dose escalation portion of the study is further designed to assess safety and tolerability, assess antitumor activity at the putative RP2D, and characterize IMGN779 PK. The data obtained during the dose escalation phase helps refine RP2D.

Secondary goals include assessment of IMGN779 safety and tolerability, characterization of IMGN779 PK profile, assessment of potential immunogenicity, and assessment of antitumor activity. Secondary endpoints include adverse reactions that occurred during treatment, PK parameters such as Cmax, AUG, and terminal half-life (t ^1/2 ), human anti-drug antibody levels, objective response rate (ORR), Includes identification of overall survival, recurrence-free survival (RFS), event-free survival (EFS), and cumulative recurrence rates at 6 and 12 months (remission period).

The observation period extends from the time the patient receives the first dose of IMGN779 until the final follow-up visit. Patients will continue to receive IMGN779 until either unacceptable toxicity or withdrawal of consent comes first, or until the drug sponsor completes the study. Patients who discontinue the study drug for reasons other than progressive disease are followed until either progressive disease, initiation of new anticancer therapy, or death first occurs.

PK parameters such as Cmax, Tmax, AUC, t1/2 (β), volume of distribution (Vss) at steady state, clearance (CL) were measured using IMGN779, total Z4681A, using actual sampling times when possible. Derivable as feasible from plasma concentrations of antibody and DGN462, and potential metabolites. If there is no actual sampling time, use the nominal time. Concentration data and PK parameters are descriptively summarized by dose. Non-compartmental methods are used to calculate PK parameters using SAS and/or Phoenix WinNonlin software. Plasma concentration time profiles are represented graphically by dose level.

Blood samples will be taken to assess the PK profile of IMGN779 when IV is administered biweekly (Schedule A) or weekly (Schedule B). Blood samples for PK measurements will be collected for all patients at the following time points and at unscheduled, end of study, and follow-up visits on Day 30:
Schedule A (Q2W)
● Cycles 1 and 3
○ Day 1-Before administration of IMGN779 and immediately after completion of infusion (+5 minutes); and 2 hours (±10 minutes), 4 hours (±15 minutes), and 6 hours (±1 hour) after completion of IMGN779 infusion. Day 2-24 hours after completion of IMGN779 infusion (±2 hours)
3rd day-48 hours after the completion of IMGN779 injection (±4 hours)
O Day 8-single blood sample o Day 15-before administration of IMGN779 and immediately after completion of infusion (+5 minutes).
● Cycle 2 and cycles 4-6
○ Day 1-Before administration of IMGN779 and immediately after completion of infusion (+5 minutes)
Day 15-Before IMGN779 administration and immediately after infusion completion (+5 minutes).
Schedule B (QW)
● Cycle 1
○ Day 1-Before administration of IMGN779 and immediately after completion of infusion (+5 minutes); and 2 hours (±10 minutes), 4 hours (±15 minutes), and 6 hours (±1 hour) after completion of IMGN779 infusion. Day 2-24 hours after completion of IMGN779 infusion (±2 hours)
3rd day-48 hours after the completion of IMGN779 injection (±4 hours)
Day 8-Before IMGN779 administration and immediately after infusion completion (+5 minutes).
Day 15-Before IMGN779 administration and immediately after infusion completion (+5 minutes).
○ Day 22-Before administration of IMGN779 and immediately after completion of infusion (+5 minutes).
● Cycle 2-6
○ Day 1-Before administration of IMGN779 and immediately after completion of infusion (+5 minutes)
Day 8-Before IMGN779 administration and immediately after infusion completion (+5 minutes).
Day 15-Before IMGN779 administration and immediately after infusion completion (+5 minutes).
○ Day 22-Before administration of IMGN779 and immediately after completion of infusion (+5 minutes).

Example 15. IMGN779 PK Exposure Metrics 19 patients with a median age of 62 received 6 dose levels ranging from 0.02 to 0.39 mg/kg IMGN779. Pharmacokinetic (PK) parameters were determined by non-compartmental analysis (NCA) of plasma samples taken at the time points outlined above in Example 14.

Adverse events (AEs) were as expected in this refractory AML population, including cytopenias and constitutional symptoms. No association between frequency or severity and IMGN779 dose level was observed. The most common AEs were nausea (41%), febrile neutropenia (29%), and rash (29%). Pneumonia, respiratory failure, and constipation were additional AEs reported in 4 or more patients (>24%). The most common serious adverse events (SAEs) were grade 3 febrile neutropenia (29%) and pneumonia (24%). No dose limiting toxicity was reported. Importantly, no safety signals for liver or hematopoietic toxicity were observed in the laboratory evaluation.

In a continuing clinical trial, a total of 23 patients with a median age of 68 years (including the 19 patients mentioned above) received 7 dose levels ranging from 0.02-0.54 mg/kg IMGN779. ..

Demographics of these 23 patients are shown in FIG. Adverse reactions that occurred during treatment are shown in Figures 17 and 18. The most frequent adverse events are:
Cytopenia: febrile neutropenia (39%), anemia (17%), and thrombocytopenia (13%);
● Gastrointestinal: constipation (22%), diarrhea (22%), and abdominal pain (17%); and ● Respiratory: dyspnea (22%), pneumonia (22%), and respiratory failure (17%).

The most frequent Grade 3 or higher adverse events are:
● Cytocytopenia: febrile neutropenia (39%), anemia (17%), and thrombocytopenia (9%); and ● Respiratory: pneumonia (22%) and respiratory failure (17%).

Other adverse events of interest included fluid response (4%), bilirubin (TBlii) elevation (8%), and alanine transaminase (ALT) elevation (4%). The most frequent serious adverse events (SAEs) included febrile neutropenia (39%), pneumonia (17%), and respiratory failure (17%). Grade 3 fluid response was a related serious adverse event.

No dose limiting toxicity was observed up to a dose of 0.54 mg/kg (cohort 7). In addition, no evidence of cumulative toxicity was observed up to a maximum of 17 doses given per patient.

Two of the three patients treated with 0.54 mg/kg IMGN779 (cohort 7) cleared peripheral blasts and the third showed a 66% reduction in peripheral blasts. One patient (cohort 6) treated with 0.39 mg/kg had less than 50% myeloblast reduction from 54% to 25% after one cycle.

The additional anti-leukemic activity observed in cohorts 6 and 7 is shown in Figures 19 and 20. FIG. 19 shows the effect of IMGN779 on peripheral hemoblasts. All patients in cohorts 6 and 7 showed a decrease from baseline in peripheral hemoblasts within the first 3-8 days of the first dose. Baseline is Peripheral Hemoblast Count on Day 0 (ie, pre-dose) for each patient. Of the 3 patients with peripheral hemoblast progression after 3-8 days, all showed a reduction in blasts after the second dose (day 15).

Figure 20 shows the effect of IMGN779 on myeloblasts. Three patients from cohorts 5 and 6 showed at least 45% reduction from baseline in myeloblasts (no hydroxyurea). One patient fell from 90% to 4% (96% below baseline). Another patient fell from 54% to 22% (59% below baseline) and a third from 23% to 12% (48% below baseline). Baseline is the number of myeloblasts on day 0 (ie, pre-dose) for each patient.

Pharmacokinetic (PK) parameters are reported for Cycle 1 of the IMGN779 Phase 1 trial (first dosing cycle per patient only) (see Figures 21A-21C and Table 19). Exposure of IMGN779 shows that data for calculating the elimination half-life of the terminal phase are minimal at inadequate low doses. IMGN779 exposure tends to increase at higher doses, with a mean (±SD) half-life of 18.3±10.2 hours at the 0.26 mg/kg dose level. Overall, exposure (AUC) and Cmax were shown to generally increase at higher doses, with Cmax and dose being linearly related at doses above 0.16 mg/kg and exposure and dose being non-linear. Yes (see Figures 21B and 21C). IMGN779 is well tolerated (at doses of 0.26 mg/kg and 0.39 mg/kg, respectively) in cohorts 5 and 6. All three patients in cohort 6 sustained the observed exposure throughout 168 hours post-infusion and sustained reduction in CD33 levels over 8 days.

The pharmacokinetic and pharmacodynamic parameters of patients in cohorts 4-7 are shown in FIG. These results show that Cmax and AUG increased with dose, with cohorts 6 and 7 showing long-term exposure over the 7 days following infusion. The duration of CD33 saturation increased with dose, and cohorts 6 and 7 showed long-term CD33 saturation over the 7 days following infusion.

The IMGN779 pharmacokinetics will continue to be evaluated and will characterize any association between clinical activity and IMGN779 pharmacokinetics.

Using a pharmacodynamic (PD) assay to measure saturation of residual free CD33, cohort 5 was shown to be consistent with saturation of free CD33 in all patients after 48 hours, consistent with PK results. It was

Without wishing to be bound by theory, at similar doses below 0.26 mg/kg, gemtuzumab ozogamicin (Mylotarg®) exposure, as indicated by the calculated Cmax. Is higher than IMGN779, suggesting that the difference may be the Kd of the antibody (ie, the Z4681A antibody contained in IMGN779 is more CD33 than the hP67.6 antibody contained in gemtuzumab ozogamicin. Had a high affinity for).

Example 16. IMGN779 Exposure and Efficacy In a sustained clinical trial, a total of 36 patients with a median age of 67 (including the 23 patients mentioned above) were dose levels ranging from 0.02 to 0.91 mg/kg IMGN779. Received.

A continuous increase in maximum concentration and exposure (AUC _last ) was observed up to a dose of 0.91 mg/kg Q2W, and the duration of increasing CD33 saturation also continued to be observed as the dose was increased (FIG. 23). ..

In addition to biweekly (Q2W) administration, IMGN779 was also administered once weekly (QW) at a dose of 0.39 mg/kg. Although once weekly administration increased the duration of CD33 saturation compared to Q2W administration, no substantial difference in IMGN779 accumulation was observed in weekly administration compared to Q2W administration ( Figure 24).

In addition, 16 of the 17 evaluable patients showed a reduction in peripheral hemoblasts within 10 days of the first dose. The median maximum reduction was 71%, ranging from 16% to 100%. In addition, 7 out of 17 evaluable patients showed a 48% to 96% reduction in myeloblasts (Figure 25). Of these 7 patients, 6 received prior intensive care, 3 had primary refractory disease, 3 had mutations in RAS, and 2 had TP53. , 2 had a mutation in IDH, and 2 had a mutation in FLT3. In addition, one patient treated with a low dose (0.16 mg/kg Q2W) showed a 93% reduction in myeloblasts (48% to 3%), prolonging treatment (cycle 5), Continued throughout cycle 14. Therefore, IMGN779 reduces peripheral hemoblasts in AML patients.

Example 17. Ongoing Assessment of Safety, Exposure Index, and Efficacy of IMGN779 As the clinical study progresses, three patients received 1.2 mg/kg IMGN779 biweekly (Q2W) and three others Patients were administered IMGN779 at 1.5 mg/kg on the same Q2W schedule. Six patients were also administered weekly (QW) with 0.54 mg/kg IMGN779.

No DLT was observed in any of the patients enrolled in this study (including the patients described in Examples 15 and 16) and no pattern of IMGN779 toxicity appeared. As shown in Figure 26, less than 40% of patients had febrile neutropenia. In addition, only one patient experienced Grade 3+ TEAE (hyperbilirubinemia) and only two patients had suspected and unexpected serious adverse events (SUSAR) (1 due to Grade 3 fluid response). 1 experienced a colon perforation). Data on hepatotoxicity are poorly corroborated, with 2 patients showing an increase in grade 3 total bilirubin (TBili) and 1 patient showing an increase in grade 3 alanine transaminase (ALT). The patient presented with grade 3 aspartate transaminase (AST). Each of these grade 3 events was observed in different patients, and the high law case was not observed based on the drug-induced liver injury assessment.

No pattern of neutropenia or thrombocytopenia was observed over the various patient cohorts, although some patients who received 0.54 mg/kg IMGN779 on the QW schedule showed cytopenias. In particular, one patient showed a significant reduction in neutrophils, hemoglobin, and platelets during cycle 1 in the SAE setting for colonic diverticulosis. The SAE resolved and patients showed a decrease in bone marrow (BM) blasts (-78%, 27% to 6%), but the number of patients did not recover during the 4 week observation. It was unclear whether the cytopenia observed in this patient was an effect of treatment with IMGN779. In addition, three other patients showed moderate to severe reductions in neutrophil and/or platelet counts, but all of these patients had progressive AML as a common confounding factor. Also, one patient had a moderate increase in platelets and a stable neutrophil count in the setting of a slight decrease in BM blasts (-30%, 26% to 18%). Collectively, none of these observations met the definition of DLT.

Pharmacokinetic and pharmacodynamic data for IMGN779 concentration and CD33 saturation were collected throughout the study. As shown in FIG. 27A, the maximum concentration of IMGN779 and exposure continued to increase with dose in cycle 1 of treatment. As shown in Figure 27B, upon repeated dosing, the QW dosing schedule was associated with a slightly higher end-of-infusion concentration compared to the Q2W dosing schedule, but without substantial accumulation of IMGN779. As shown in FIGS. 28A and 28B, increased CD33 saturation (ie, availability of low CD33 receptors on blasts) was observed with increasing dose. Saturation was not maintained between doses of Q2W schedule (Figure 28A), while saturation was maintained between doses of QW schedule (Figure 28B). As shown in FIGS. 29A and 29B, complete saturation occurred at 0.54 mg/kg QW administration (FIG. 29B), whereas incomplete saturation was observed at the same dose of Q2W administration (FIG. 29A). Taken together, these data show that exposure indicators (eg, Cmax and AUC) increase with dose and that CD33 saturation persists with the QW dosing schedule.

To assess whether the CD33 SNPs are associated with response to IMGN779 treatment, the level of CD33 expression on patient cells is determined and further characterized by the combination of SNP alleles. As shown in Figures 30A and 30B, uniform CD33 expression across blasts appeared to be a better indicator of response than the amount of CD33 on each blast. In addition, the majority of informal responders had high CD33% positivity. Previous studies with gemtuzumab ozogamicin have suggested specific therapeutic benefit in pediatric AML patients with the rs12459419 SNP (associated with the CC allele); however, the same trend is associated with conventional MRC adult AML clinical It was not observed in the test. In this study with adult AML patients, the clinical benefit of IMGN779 was observed with all three possible SNP allele combinations (CC, CT, and TT), as shown in FIG. Thus, these data suggest that the CD33 T allele does not interfere with the response to IMGN779 and that the particular CD33 SNP does not provide a definitive predictor of response.

Finally, the best response (measured as% reduction in myeloblasts) was observed across all patient cohorts in the study. Only one patient had a complete response (CRi) with inadequate recovery of hematopoietic function, but a significant proportion of patients (<44%, 11 out of 25) showed signs of efficacy. Indicated. As shown in Figure 32, efficacy signals were seen for both Q2W and QW dosing schedules. Taken together, these data suggest a broad activity of IMGN779 in the relapsed/refractory AML population.

Example 18. Pharmacokinetic Modeling of IMGN779 Pharmacokinetic (PK) modeling was used to assess the effect of IMGN779 dose on elimination half-life. A clear dose effect on elimination half-life was observed in patients who received IMGN779 and a slow elimination half-life was found at high doses. In particular, IMGN779 blood levels were rapidly reduced to 10-20 nM in all low dose subjects. The same rapid decline was also observed in some individuals who received high doses, followed by a slow elimination phase. The rapid elimination of IMGN779 observed in low dose subjects (ie, resulting in a final IMGN779 blood concentration of <10 nM) but not in the highest dose subjects suggests an inconsistent non-linear concentration range. did. A simple one-compartment PK model was then applied to the probe data to estimate individual concentration-time slopes. As shown in FIG. 33, the dose dependency was confirmed by the slope estimation value. However, as shown in FIG. 34, the conventional linear PK model that did not consider target or dose changes did not fit the data well.

Data from patients receiving 0.54 mg/kg IMGN779 were then used to assess the correlation between the elimination rate constant and the number of binding sites per cell and total target concentration. As shown in Figures 35A and 35B, the elimination rate constants did not correlate with the number of binding sites (Figure 35A) or total target concentration (Figure 35B). The target-mediated pharmacokinetics (TMDD) model was then used to assess linear + target-mediated elimination. Since only total target (R+RL) was measured, instantaneous binding was assumed. The number of binding sites per cell (ABC) was determined for monocytes, lymphocytes, blasts, and granulocytes to calculate the targets of measurement. Targets of measurement (ie ABC x cell count (n/L)) were calculated in circulating cells by cell type and summed. As shown in Figure 36, TMDD based on measured levels did not fit better than the linear PK model. Therefore, similar to the linear model, the addition of the target-mediated elimination model with the measurement target could not explain the observed PK profile.

A simple binding model was then applied to the data. Among them, the estimated target level was independent of the target level and there was no target-mediated efflux (saturated binding only). The target concentration pool was estimated at 16.4 nM and compared to the measured concentration value of 0.02 nM. As shown in FIGS. 37A and 37B, no clear trend was observed between putative target concentrations and individually measured circulating target concentrations, and the pool of target/non-specific binding required to explain PK was It was suggested to be outside the blood. Although not a perfect fit, this combined model was associated with an improved fit to the data as compared to the linear PK or TMDD model with the measurement target (Figure 38).

Claims (60)

The following formula:

(I) about 0.54 to about 0.8 mg/kg, (ii) about 0.30 to about 0.54 mg/kg, (iii) about an anti-CD33 immunoconjugate or a pharmaceutically acceptable salt thereof. 0.7 to about 1 mg/kg, (iv) about 0.39 to about 0.7 mg/kg, (v) about 0.7 to about 1.25 mg/kg or (vi) about 1 to about 2 mg/kg. A method of treating cancer in a subject, comprising administering to the subject,
In the formula:
Double line between N and C

Represents a single bond or a double bond, provided that when it is a double bond, X is absent and Y is hydrogen; when it is a single bond, X is hydrogen and Y is There in the -SO ₃ H;
r is an integer of 1 to 10; and A is a variable heavy chain containing the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 1 to 3, and CDR1 and CDR2 set forth in SEQ ID NOs: 4 to 6, respectively. And a variable light chain comprising a CDR3 sequence, which is an anti-CD33 antibody or an antigen-binding fragment thereof. The method of claim 1, wherein about 0.54 to about 0.70 mg/kg of the immunoconjugate is administered. The method of claim 1, wherein about 0.54 or about 0.70 mg/kg of the immunoconjugate is administered. 4. The method of any one of claims 1-3, wherein the immunoconjugate is administered about once every two weeks. The method of claim 1, wherein about 0.39 to about 0.54 mg/kg of the immunoconjugate is administered. The method of claim 1, wherein about 0.39 mg/kg or about 0.54 mg/kg of the immunoconjugate is administered. 7. The method of any one of claims 1, 5 and 6, wherein the immunoconjugate is administered about once a week. The method of claim 1, wherein about 0.7 to about 0.9 mg/kg of the immunoconjugate is administered. Administering about 0.7 mg/kg, about 0.75 mg/kg, about 0.8 mg/kg, about 0.85 mg/kg, about 0.9 mg/kg, or about 1 mg/kg of the immunoconjugate. The method according to Item 1. 10. The method of any one of claims 1, 8 and 9, wherein the immunoconjugate is administered about once every two weeks. About 0.39 mg/kg, about 0.45 mg/kg, about 0.5 mg/kg, about 0.54 mg/kg, about 0.55 mg/kg, about 0.6 mg/kg, about 0.65 mg/kg, or The method of claim 1, wherein about 0.7 mg/kg of the immunoconjugate is administered. 12. The method of claim 1 or 11, wherein the immunoconjugate is administered about once a week. 2. The method of claim 1, wherein about 0.7 to about 1 mg/kg or about 0.7 to about 0.9 mg/kg of the immunoconjugate is administered. About 0.7 mg/kg, about 0.75 mg/kg, about 0.8 mg/kg, about 0.85 mg/kg, about 0.9 mg/kg, about 0.95 mg/kg, or about 1 mg/kg of said immuno. The method of claim 1, wherein the conjugate is administered. 15. The method of claim 1, 13 or 14 wherein the immunoconjugate is administered about once per week. The method of claim 1, wherein about 1 to about 1.25 mg/kg of the immunoconjugate is administered. The method of claim 1, wherein about 1.1 mg/kg, about 1.15 mg/kg, about 1.2 mg/kg, or about 1.25 mg/kg of the immunoconjugate is administered. 18. The method of any one of claims 1, 16 and 17, wherein the immunoconjugate is administered about once every two weeks. 18. The method of any one of claims 1, 16 and 17, wherein the immunoconjugate is administered about once per week. The method of claim 1, wherein about 1.2 to about 2 mg/kg or about 1.5 to about 1.6 mg/kg of the immunoconjugate is administered. The method of claim 1, wherein about 1.5 mg/kg or about 1.56 mg/kg of the immunoconjugate is administered. 22. The method of any one of claims 1, 20, and 21, wherein the immunoconjugate is administered about once every two weeks. 22. The method of any one of claims 1, 20, and 21, wherein the immunoconjugate is administered about once per week. 24. The method of any one of claims 1-23, wherein the immunoconjugate is administered on a 28-day cycle. 25. The method of any one of claims 1-24, wherein the immunoconjugate is administered by intravenous infusion. 26. The method of any one of claims 1-25, wherein the variable heavy chain comprises the sequence set forth in SEQ ID NO:9 and the variable light chain comprises the sequence set forth in SEQ ID NO:10. 27. The method of claim 26, wherein the antibody or antigen binding fragment thereof comprises a heavy chain comprising the sequence set forth in SEQ ID NO:11 and a light chain comprising the sequence set forth in SEQ ID NO:12. The immunoconjugate is
Formula (II):

Or an pharmaceutically acceptable salt thereof, wherein A is the anti-CD33 antibody or an antigen-binding fragment thereof, and r is an integer of 1 to 10. The method according to any one of 1. The immunoconjugate is
Formula (III):

Or an pharmaceutically acceptable salt thereof, wherein A is the anti-CD33 antibody or an antigen-binding fragment thereof, and r is an integer of 1 to 10. The method according to any one of 1. 30. The method of any one of claims 1-29, wherein the pharmaceutically acceptable salt is a sodium or potassium salt. 31. The method of any one of claims 1-30, wherein r is an integer from 2-9, 3-8, 4-7, or 5-6. The immunoconjugate is included in a composition comprising at least two of the immunoconjugates, wherein the average number of cytotoxins per antibody or antigen-binding fragment thereof is 2-8, 3-7, 3- The method according to any one of claims 1 to 30, which is 5, or 2.5 to 3.5. 33. The method of any one of claims 1-32, wherein the immunoconjugate is IMGN779. 34. The method of any one of claims 1-33, wherein the cancer is selected from the group consisting of leukemia, lymphoma, and myeloma. The cancer is acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute lymphoblastic leukemia (ALL), B cell lineage of acute lymphoblastic leukemia (B ALL), T cell acute lymphoma. Blastic leukemia (T ALL), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), myelodysplastic syndrome (MDS), basic plasmacytoid DC tumor (BPDCN) leukemia, non-Hodgkin lymphoma ( 35. The method of claim 34, wherein the method is selected from the group consisting of NHL), mantle cell lymphoma, and Hodgkin leukemia (HL). 36. The method of claim 35, wherein the cancer is acute myelogenous leukemia (AML). 37. The method of claim 36, wherein the AML is refractory or relapsed acute myelogenous leukemia. 37. The method of claim 36, wherein the AML is newly diagnosed. The AML has overexpression of P-glycoprotein, EVI1 overexpression, p53 mutation, DNMT3A mutation, FLT3 internal tandem duplication, complex karyotype, decreased expression of BRCA1, BRCA2, or PALB2, or BRCA1, BRCA2, or PALB2. 39. The method of any one of claims 36-38 characterized by the mutation of 36. The method of claim 35, wherein the MDS is a high risk MDS. 41. The method of any one of claims 1-40, wherein the homozygous rs12459419C genotype has been detected in a sample obtained from the subject. 42. The method of claim 41, further comprising detecting the homozygous rs12459419C genotype in a sample obtained from the subject. 41. The method of any one of claims 1-40, wherein a heterozygous rs12459419C genotype has been detected in a sample obtained from the subject. 44. The method of claim 43, further comprising detecting the heterozygous rs12459419C genotype in a sample obtained from the subject. 41. The method of any one of claims 1-40, wherein a homozygous rs12459419T genotype has been detected in a sample obtained from the subject. 46. The method of claim 45, further comprising detecting the homozygous rs12459419T genotype in a sample obtained from the subject. 41. The method of any one of claims 1-40, wherein a heterozygous rs12459419T genotype has been detected in a sample obtained from the subject. 48. The method of claim 47, further comprising detecting the heterozygous rs12459419T genotype in a sample obtained from the subject. 49. The method of any one of claims 41-48, wherein the sample obtained from the patient is a blood sample or a buccal swab. 50. The method of any one of claims 1-49, wherein the cancer is chemosensitive. 50. The method of claims 1-49, wherein the cancer is chemoresistant. 52. The method of any one of claims 1-51, wherein at least 20% of the cancer-derived blasts are CD33 positive, as measured by flow cytometry. 53. The method of any one of claims 1-52, wherein said administration results in saturation of free CD33. 54. The method of any one of claims 1-53, wherein the administration results in a reduction in peripheral hemoblasts. 55. The method of any one of claims 1-54, wherein the administration results in a reduction in myeloblasts. 56. The method of any one of claims 1-55, wherein the subject is a human. 57. The method of any one of claims 1-56, wherein the cancer is characterized by a RAS mutation. 58. The method of any one of claims 1-57, wherein the cancer is characterized by a TP53 mutation. 59. The method of any one of claims 1-58, wherein the cancer is characterized by an IDH mutation. 60. The method of any one of claims 1-59, wherein the cancer is characterized by a FLT3 mutation.