EFFECTIVE TARGETING OF PRIMARY HUMAN LEUKEMIA USING ANTI-CD123 CHIMERIC ANTIGEN RECEPTOR ENGINEERED T CELLS

• US 20170183415A1
• Filed: 01/06/2017
• Published: 06/29/2017
• Est. Priority Date: 02/20/2013
• Status: Active Grant

• Current Assignee: Novartis Ag, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Novartis Ag, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: June, Carl H., Brogdon, Jennifer, Gill, Saar, Kalos, Michael D., Scholler, John, Loew, Andreas
• Granted Patent: US 11,028,177 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/507 Comprising a combination of two or more separate antibodies
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 38/1774 Immunoglobulin superfamily (e.g. CD2, CD4, CD8, ICAM molecules, B7 molecules, Fc-receptors, MHC-molecules)
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/464429 Molecules with a "CD" designation not provided for elsewhere
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70517 CD8
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/53 Hinge
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/33 : fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies

Abstract

The invention provides compositions and methods for treating leukemia, for example, acute myeloid leukemia (AML) and B-cell acute lymphoid leukemia (B-ALL). The invention also relates to at least one chimeric antigen receptor (CAR) specific to CD123, vectors comprising the same, and recombinant T cells comprising the CD123 CAR. The invention also includes methods of administering a genetically modified T cell expressing a CAR that comprises a CD123 binding domain. The invention also includes methods of bone marrow ablation for use in treatments necessitating bone marrow reconditioning or transplant.

Claims

1-102. -102. (canceled)

103. A method of eradicating at least a portion of existing bone marrow in a subject, the method comprising administering to the subject an effective amount of cells expressing a chimeric antigen receptor (CAR) molecule comprising an anti-CD123 binding domain, a transmembrane domain, and an intracellular signaling domain, thereby eradicating at least a portion of existing bone marrow in a subject.

117. The method of claim 103, wherein the method is a cellular conditioning method prior to bone marrow transplantation.

118. The method of claim 103, wherein the portion of existing bone marrow eradicated comprises normal CD123-expressing bone marrow cells.

119. The method of claim 103, wherein the portion of existing bone marrow eradicated comprises CD123-expressing bone marrow myeloid progenitor cells.

120. The method of claim 103, wherein all or essentially all of the existing bone marrow in the subject is eradicated.

121. The method of claim 103, wherein the number of CD123-expressing cells in bone marrow is reduced.

122. The method of claim 103, wherein the subject is in need of a bone marrow transplant or bone marrow reconditioning.

123. The method of claim 103, wherein the subject has a disease, disorder or condition that is treatable with a bone marrow transplant or bone marrow reconditioning.

124. The method of claim 123, wherein the disease, disorder or condition is selected from the group consisting of a hematological cancer, a solid tumor, a hematological disease, a metabolic disorder, HIV, HTLV, a lysosomal storage disorder and an immunodeficiency.

125. The method of claim 124, wherein the hematological cancer is a leukemia, lymphoma, or myeloma.

126. The method of claim 124, wherein the hematological cancer acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphoid leukemia (CLL), chronic myelogenous leukemia (CML), Hodgkin'"'"'s disease, Non-Hodgkin'"'"'s lymphoma, or multiple myeloma.

127. The method of claim 103, wherein the method is performed prior to a bone marrow transplant.

128. The method of claim 103, further comprising administering a bone marrow transplant to the subject following the eradication of at least a portion of existing bone marrow in the subject.

129. The method of claim 128, wherein the bone marrow transplant comprises transplantation of a stem cell.

130. The method of claim 128, wherein the bone marrow transplant comprises transplantation of autologous bone marrow cells or autologous stem cells.

131. The method of claim 128, wherein the bone marrow transplant comprises transplantation of allogeneic bone marrow cells or allogeneic stem cells.

132. The method of claim 103, wherein the subject has a hematological cancer or a solid tumor.

133. The method of claim 132, further comprising administering one or more anti-cancer therapy.

134. The method of claim 133, wherein the anti-cancer therapy is an anti-cancer CAR therapy, chemotherapy, or radiation.

135. The method of claim 103, wherein the cells expressing the CAR molecule are T cells.

136. The method of claim 103, wherein the cells expressing the CAR molecule are CD8+ T cells.

137. The method of claim 103, wherein the cells expressing the CAR molecule are human cells.

138. The method of claim 103, wherein the cells expressing the CAR molecule are allogeneic T cells.

139. The method of claim 103, wherein the cells expressing the CAR molecule are autologous T cells.

140. The method of claim 103, wherein the anti-CD123 binding domain comprises an antibody molecule.

141. The method of claim 140, wherein the antibody molecule comprises a scFv, a F(ab′

), a F(ab′

)2, a Fv fragment, an immunoglobulin single domain antibody (sdAb), a single light chain variable domain (VL), a single heavy chain variable domain (VH), or a nanobody.

142. The method of claim 140, wherein the antibody molecule is a murine, human, or humanized antibody molecule.

143. The method of claim 103, wherein the anti-CD123 binding domain comprises one or more of light chain complementary determining region 1 (LC CDR1), light chain complementary determining region 2 (LC CDR2), and light chain complementary determining region 3 (LC CDR3) of any anti-CD123 light chain binding domain amino acid sequence listed in Table 1 or SEQ ID NO:

2 or SEQ ID NO;

101, and one or more of heavy chain complementary determining region 1 (HC CDR1), heavy chain complementary determining region 2 (HC CDR2), and heavy chain complementary determining region 3 (HC CDR3) of any anti-CD123 heavy chain binding domain amino acid sequence listed in Table 1 or SEQ ID NO;

2 or SEQ ID NO;

101.

144. The method of claim 103, wherein said anti-CD123 binding domain comprises:

(a) a light chain variable region comprising;

a light chain complementarity determining region 1 (LC CDR1) comprising the sequence of SEQ ID NO;

20, a light chain complementarity determining region 2 (LC CDR2) comprising the sequence of SEQ ID NO;

21, and a light chain complementarity determining region 3 (LC CDR3) comprising the sequence of SEQ ID NO;

22, and

a heavy chain variable region comprising;

a heavy chain complementarity determining region 1 (HC CDR1) comprising the sequence of SEQ ID NO;

16, a heavy chain complementarity determining region 2 (HC CDR2) comprising the sequence of SEQ ID NO;

17, and a heavy chain complementarity determining region 3 (HC CDR3) comprising the sequence of SEQ ID NO;

18;

or

(b) a light chain variable region comprising;

a light chain complementarity determining region 1 (LC CDR1) comprising the sequence of SEQ ID NO;

28, a light chain complementarity determining region 2 (LC CDR2) comprising the sequence of SEQ ID NO;

29, and a light chain complementarity determining region 3 (LC CDR3) comprising the sequence of SEQ ID NO;

30, and

a heavy chain variable region comprising;

a heavy chain complementarity determining region 1 (HC CDR1) comprising the sequence of SEQ ID NO;

24, a heavy chain complementarity determining region 2 (HC CDR2) comprising the sequence of SEQ ID NO;

25, and a heavy chain complementarity determining region 3 (HC CDR3) comprising the sequence of SEQ ID NO;

26;

or

(c) a light chain variable region comprising;

a light chain complementarity determining region 1 (LC CDR1) comprising the sequence of SEQ ID NO;

87, a light chain complementarity determining region 2 (LC CDR2) comprising the sequence of SEQ ID NO;

88, and a light chain complementarity determining region 3 (LC CDR3) comprising the sequence of SEQ ID NO;

89, and

a heavy chain variable region comprising;

a heavy chain complementarity determining region 1 (HC CDR1) comprising the sequence of SEQ ID NO;

84, a heavy chain complementarity determining region 2 (HC CDR2) comprising the sequence of SEQ ID NO;

85, and a heavy chain complementarity determining region 3 (HC CDR3) comprising the sequence of SEQ ID NO;

86.

145. The method of claim 103, wherein

(a) the anti-CD123 binding domain comprises a light chain variable region comprising an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications of an amino acid sequence of a light chain variable region provided in SEQ ID NO:

36, SEQ ID NO;

42, SEQ ID NO;

48, SEQ ID NO;

54, SEQ ID NO;

60, SEQ ID NO;

66, SEQ ID NO;

72, or SEQ ID NO;

78, or a sequence with 95-99% identity to an amino acid sequence provided in SEQ ID NO;

36, SEQ ID NO;

42, SEQ ID NO;

48, SEQ ID NO;

54, SEQ ID NO;

60, SEQ ID NO;

66, SEQ ID NO;

72, or SEQ ID NO;

78;

(b) the anti-CD123 binding domain comprises a heavy chain variable region comprising an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications of an amino acid sequence of a heavy chain variable region provided in SEQ ID NO;

36, SEQ ID NO;

42, SEQ ID NO;

48, SEQ ID NO;

54, SEQ ID NO;

60, SEQ ID NO;

66, SEQ ID NO;

72, or SEQ ID NO;

78, or a sequence with 95-99% identity to an amino acid sequence provided in SEQ ID NO;

36, SEQ ID NO;

42, SEQ ID NO;

48, SEQ ID NO;

54, SEQ ID NO;

60, SEQ ID NO;

66, SEQ ID NO;

72, or SEQ ID NO;

78;

or

(c) both (a) and (b).

146. The method of claim 103, wherein the anti-CD123 binding domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

36, SEQ ID NO;

42, SEQ ID NO;

48, SEQ ID NO;

54, SEQ ID NO;

60, SEQ ID NO;

66, SEQ ID NO;

72, and SEQ ID NO;

78, or a sequence with 95-99% identity thereof.

147. The method of claim 103, wherein the anti-CD123 binding domain comprises the amino acid sequence of SEQ ID NO:

101 or SEQ ID NO;

2, or an amino acid sequence with 95-99% identity thereto.

148. The method of claim 103, wherein the transmembrane domain comprises a transmembrane domain of a protein selected from the group consisting of the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 and CD154.

149. The method of claim 103, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:

5;

an amino acid sequence comprising at least one, two or three modifications but not more than 20, 10 or 5 modifications of the amino acid sequence of SEQ ID NO;

5;

or an amino acid sequence with 95-99% identity to the amino acid sequence of SEQ ID NO;

5.

150. The method of claim 103, wherein the anti-CD123 binding domain is connected to the transmembrane domain by a hinge region.

151. The method of claim 150, wherein the hinge region comprises the amino acid sequence of any of SEQ ID NO:

4, 104, 122 or 124, or a sequence with 95-99% identity thereof.

152. The method of claim 103, wherein the intracellular signaling domain comprises a functional signaling domain of 4-1BB and a functional signaling domain of CD3 zeta.

153. The method of claim 103, wherein the intracellular signaling domain comprises a costimulatory domain.

154. The method of claim 103, wherein the intracellular signaling domain comprises a primary signaling domain.

155. The method of claim 103, wherein the intracellular signaling domain comprises a costimulatory domain and a primary signaling domain.

156. The method of claim 103, wherein the intracellular signaling domain comprises:

(i) the amino acid sequence of SEQ ID NO;

6 or SEQ ID NO;

23;

(ii) the amino acid sequence of SEQ ID NO;

7 or SEQ ID NO;

98;

or

(iii) both (i) and (ii).

157. The method of claim 153, wherein the costimulatory domain comprises a functional signaling domain of a protein selected from the group consisting of OX40, CD27, CD28, CD5, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), and 4-1BB (CD137).

158. The method of claim 153, wherein the costimulatory domain comprises

(i) the amino acid sequence of SEQ ID NO:

6 or SEQ ID NO;

23;

an amino acid sequence comprising at least one, two or three modifications but not more than 20, 10 or 5 modifications of the amino acid sequence of SEQ ID NO;

6 or SEQ ID NO;

23;

or a sequence with 95-99% identity to the amino acid sequence of SEQ ID NO;

6 or SEQ ID NO;

23.

159. The method of claim 103, wherein intracellular signaling domain comprises:

(i) a costimulatory domain comprising a functional signaling domain of a protein chosen from OX40, CD27, CD28, CD5, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), or 4-1BB (CD137);

(ii) a primary signaling domain comprising a functional signaling domain derived from CD3 zeta, FcR gamma, FcR beta, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, CD278, CD66d, DAP10, or DAP12;

or

(iii) both (i) and (ii).

160. The method of claim 155, wherein the costimulatory domain comprises the amino acid sequence of SEQ ID NO:

6 or SEQ ID NO;

23;

an amino acid sequence having at least one, two or three modifications but not more than 20, 10 or 5 modifications of the amino acid sequence of SEQ ID NO;

6 or SEQ ID NO;

23;

or a sequence with 95-99% identity to the amino acid sequence of SEQ ID NO;

6 or SEQ ID NO;

23, and the primary signaling domain comprises the amino acid sequence of SEQ ID NO;

7 or SEQ ID NO;

98;

an amino acid sequence having at least one, two or three modifications but not more than 20, 10 or 5 modifications of the amino acid sequence of SEQ ID NO;

7 or SEQ ID NO;

98;

or a sequence with 95-99% identity to the amino acid sequence of SEQ ID NO;

7 or SEQ ID NO;

98.

161. The method of claim 103, wherein the CAR molecule comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

41, SEQ ID NO;

47, SEQ ID NO;

53, SEQ ID NO;

59, SEQ ID NO;

65, SEQ ID NO;

71, SEQ ID NO;

77, or SEQ ID NO;

83, or a sequence with 95-99% identity thereof.

162. The method of claim 103, wherein the CAR molecule comprises an amino acid sequence of SEQ ID NO:

100, or a sequence with 95-99% identity thereof.

163. The method of claim 103, wherein the CAR molecule comprises an amino acid sequence of SEQ ID NO:

97, or a sequence with 95-99% identity thereof.

104-114. -114. (canceled)

115. A method of treating a subject having cancer, the method comprising

administering to the subject an amount of cells expressing a chimeric antigen receptor (CAR) molecule effective to eradicate at least a portion of existing bone marrow in a subject, wherein said CAR molecule comprises an anti-CD123 binding domain, a transmembrane domain, and an intracellular signaling domain,

thereby treating the subject having cancer.

116. A method of treating a subject having a disease, disorder or condition that is treatable with a bone marrow transplant or bone marrow reconditioning, the method comprising

administering to the subject an amount of cells expressing a chimeric antigen receptor (CAR) molecule effective to eradicate at least a portion of existing bone marrow in a subject, wherein said CAR molecule comprises an anti-CD123 binding domain, a transmembrane domain, and an intracellular signaling domain,

thereby treating the subject having a disease, disorder or condition that is treatable with a bone marrow transplant or bone marrow reconditioning.