# US 12,006,370 B2 · Use of anti-CD5 antibody drug conjugate (ADC) in allogeneic cell therapy

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 12,006,370 B2
- Document ID: US12006370B2
- Status: Granted
- Kind: B2
- Application number: 17/155,832
- Filed: 2021-01-22
- Published: 2021-11-18
- Granted: 2024-06-11
- Earliest claimed: 2018-07-23
- Assignees: Heidelberg Pharma Research GmbH
- Applicants: Heidelberg Pharma Research GmbH
- Inventors: Anthony Boitano; Michael Cooke
- Web page: https://patentgrep.com/patent/US12006370B2

## CPC

- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- C07K 16/2896: {against molecules with a "CD"-designation, not provided for elsewhere}
- A61K 31/40: having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K 31/535: having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K 31/5517: condensed with five-membered rings having nitrogen as a ring hetero atom, e.g. imidazobenzodiazepines, triazolam
- A61K 31/704: attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin {(digitoxin A61K31/7048)}
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/22: Immunosuppressive or immunotolerising
- A61K 40/31: Chimeric antigen receptors [CAR]
- A61K 40/418: {Antigens related to induction of tolerance to non-self}
- A61K 47/6809: {Antibiotics, e.g. antitumor antibiotics anthracyclins, adriamycin, doxorubicin or daunomycin}
- A61K 47/6831: {Fungal toxins, e.g. alpha sarcine, mitogillin, zinniol or restrictocin}
- A61K 47/6849: {the antibody targeting a receptor, a cell surface antigen or a cell surface determinant}
- A61P 35/00: Antineoplastic agents
- A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection
- C07K 14/7051: {T-cell receptor (TcR)-CD3 complex}
- C07K 14/70521: {CD28, CD152}
- C07K 14/70578: {NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 (NGF-receptor C07K14/71, TNF-receptor C07K14/7151)}
- C07K 16/2806: {against CD2}
- C12N 5/0087: {Purging against subsets of blood cells, e.g. purging alloreactive T cells}
- C12N 5/0636: {T lymphocytes}
- A61K 40/50: characterised by the use of allogeneic cells
- A61K 2239/31: characterized by the route of administration
- A61K 2239/38: characterised by the dose, timing or administration schedule
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- C07K 2319/02: containing a signal sequence
- C12N 2510/00: Genetically modified cells
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The invention provides methods of depleting CD5+ cells in human patients undergoing chimeric antigen receptor (CAR) immunotherapy in order to promote acceptance of CAR expressing immune cells. Anti-CD5 antibody drug conjugates (ADCs) are administered as a conditioning regimen to a human patient receiving autologous or allogeneic CAR expressing immune cells such that the CAR expressing immune cells are accepted by the human patient. Compositions and methods of the invention can be used in combination with CAR therapy to treat a variety of pathologies, including autoimmune diseases and cancer.

## Claims

### Claim 1

A method of promoting acceptance of an immune cell expressing a chimeric antigen receptor (CAR) in a human subject having cancer or an autoimmune disease, the method comprising:
(a) administering an anti-CD5 antibody drug conjugate (ADC) to a human subject having cancer or an autoimmune disease, wherein the anti-CD5 ADC comprises an anti-CD5 antibody, or antigen-binding fragment thereof, conjugated to a cytotoxin via a linker; and
(b) after administering said anti-CD5 antibody drug conjugate, administering a therapeutically effective amount of an immune cell expressing a CAR to the human subject, wherein the CAR comprises an extracellular domain that binds to a tumor antigen or an antigen associated with an autoimmune disease, a transmembrane domain, and a cytoplasmic domain.

### Claim 2 (depends on 1)

The method of claim 1, wherein the human subject is not administered alemtuzumab and/or a lymphodepleting chemotherapeutic agent prior to, concomitantly with, or following step (b).

### Claim 3 (depends on 2)

The method of claim 2, wherein said human subject is not administered a lymphodepleting chemotherapeutic agent, the lymphodepleting chemotherapeutic agent is selected from the group consisting of fludarabine, cyclophosphamide, bendamustine, and pentostatin.

### Claim 4 (depends on 1)

The method of claim 1, further comprising administering an anti-CD2 ADC to the human subject prior to step (b).

### Claim 5 (depends on 1)

The method of claim 1, wherein the immune cell is an allogeneic cell or an autologous cell.

### Claim 6 (depends on 5)

The method of claim 5, wherein said immune cell is an allogeneic cell is selected from the group consisting of an allogeneic T cell and an allogeneic NK cell.

### Claim 7

A method of treating a patient having a tumor, comprising:
administering to a subject in need thereof:
(i) an anti-CD5 ADC, wherein the anti-CD5 ADC comprises an anti-CD5 antibody, or antigen-binding fragment thereof, conjugated to a cytotoxin via a linker, and
(ii) after administering said anti-CD5 ADC, administering to the patient a therapeutically effective amount of from about 1×106 to about 1×108 engineered CAR T cells/kg.

### Claim 8 (depends on 1)

The method of claim 1, wherein the anti-CD5 ADC is administered to the patient as a single dose or as multiple doses.

### Claim 9 (depends on 1)

The method of claim 1, wherein the anti-CD5 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 having an amino acid sequence as set forth in SEQ ID NOs: 3, 4, and 5, respectively, and comprises a light chain variable region comprising a CDR1, a CDR2, and a CDR3 having an amino acid sequence as set forth in SEQ ID NOs: 6, 7, and 8, respectively.

### Claim 10 (depends on 1)

The method of claim 1, wherein the cytotoxin is an antimitotic agent or an RNA polymerase inhibitor.

### Claim 11 (depends on 10)

The method of claim 10, wherein said cytotoxin is an RNA polymerase inhibitor which is an amatoxin.

### Claim 12 (depends on 11)

The method of claim 11, wherein the anti-CD5 antibody, or antigen-binding fragment thereof conjugated to an amatoxin is represented by the formula Ab-Z-L-Am, wherein Ab is the antibody or antigen—binding fragment thereof, L is a linker, Z is a chemical moiety, and Am is an amatoxin represented by formula (III)
wherein R1 is H, OH, ORA, or ORC;
R2 is H, OH, ORB, or ORC;
RA and RB, when present, together with the oxygen atoms to which they are bound, combine
to form an optionally substituted 5-membered heterocycloalkyl group;
R3 is H, RC, or RD;
R4, R5, R6, and R7 are each independently H, OH, ORC, ORD, RC, or RD;
R8 is OH, NH2, ORC, ORD, NHRC, or NRCRD;
R9 is H, OH, ORC, or ORD;
Q is —S—, —S(O)—, or —SO2—;
RC is -L-Z;
RD is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally
substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, optionally substituted C2-C6
alkynyl, optionally substituted C2-C6 heteroalkynyl, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl;
L is a linker; and
Z is a chemical moiety formed from a coupling reaction between a reactive substituent present on L and a reactive substituent present within the anti-CD5 antibody or antigen-binding fragment thereof, wherein Am comprises exactly one RC substituent.

### Claim 13 (depends on 10)

The method of claim 10, wherein said cytotoxin is at least one antimitotic agent selected from the group consisting of a maytansine, an auristatin, a pyrrolobenzodiazepine (PBD), and a calicheamicin.

### Claim 14 (depends on 11)

The method of claim 11, wherein the linker of the ADC is N-beta-maleimidopropionyl-Val-Ala-para-aminobenzyl (BMP-Val-Ala-PAB).

### Claim 15 (depends on 1)

The method of claim 1, wherein the extracellular domain of the CAR comprises an scFv antibody or a single chain T cell receptor (scTCR).

### Claim 16 (depends on 1)

The method of claim 1, wherein the tumor antigen is an antigen selected from the group consisting of CD19, CD22, CD30, CD7, BCMA, CD137, CD22, CD20, AFP, GPC3, MUC1, mesothelin, CD38, PD1, EGFR, MG7, BCMA, TACI, CEA, PSCA, CEA, HER2, MUC1, CD33, ROR2, NKR-2, PSCA, CD28, TAA, NKG2D, or CD123.

### Claim 17 (depends on 1)

The method of claim 1, wherein the cytoplasmic domain of the CAR comprises a CD28 cytoplasmic signaling domain, a CD3 zeta cytoplasmic signaling domain, an OX40 cytoplasmic signaling domain, and/or a CD137 (4-1BB) cytoplasmic signaling domain.

### Claim 18 (depends on 1)

The method of claim 1, wherein the cytoplasmic domain of the CAR comprises a CD3 zeta cytoplasmic signaling domain.

### Claim 19 (depends on 1)

The method of claim 1, wherein the human subject having cancer has a cancer selected from the group consisting of leukemia, adult advanced cancer, pancreatic cancer, non-resectable pancreatic cancer, colorectal cancer, metastatic colorectal cancer, ovarian cancer, triple-negative breast cancer, hematopoietic/lymphoid cancer, colon cancer liver metastasis, small cell lung cancer, non-small cell lung cancer, B-cell lymphoma, relapsed or refractory B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large cell lymphoma, relapsed or refractory diffuse large cell lymphoma, anaplastic large cell lymphoma, primary mediastinal B-cell lymphoma, recurrent mediastinal, refractory mediastinal large B-cell lymphoma, large B-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, relapsed or refractory non-Hodgkin lymphoma, refractory aggressive non-Hodgkin lymphoma, B-cell non-Hodgkin lymphoma, refractory non-Hodgkin lymphoma, colorectal carcinoma, gastric carcinoma, pancreatic carcinoma, triple-negative invasive breast carcinoma, renal cell carcinoma, lung squamous cell carcinoma, hepatocellularcarcinoma, urothelial carcinoma, leukemia, B-cell leukemia, B-cell acute lymphocytic leukemia, B-cell acute lymphoblastic leukemia, adult acute lymphoblastic leukemia, B-cell prolymphocytic leukemia, childhood acute lymphoblastic leukemia, refractory childhood acute lymphoblastic leukemia, acute leukemia, acute lymphoblastic leukemia, acute lymphocytic leukemia, prolymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, recurrent plasma cell myeloma, refractory plasma cell myeloma, multiple myeloma, relapsed or refractory multiple myeloma, multiple myeloma of bone, malignant glioma of brain, myelodysplastic syndrome, EGFR-positive colorectal cancer, glioblastoma multiforme, neoplasms, blastic plasmacytoid dendritic cell neoplasms, liver metastases, solid tumors, advanced solid tumors, mesothelin positive tumors, and hematological malignancies.

### Claim 20 (depends on 7)

The method of claim 7, wherein the anti-CD5 ADC is administered to the patient as multiple doses.

### Claim 21 (depends on 7)

The method of claim 7, wherein the anti-CD5 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 having an amino acid sequence as set forth in SEQ ID NOs: 1, 2, and 3, respectively, and comprises a light chain variable region comprising a CDR1, a CDR2, and a CDR3 having an amino acid sequence as set forth in SEQ ID NOs: 4, 5, and 6, respectively.

### Claim 22 (depends on 7)

The method of claim 7, wherein the cytotoxin is an antimitotic agent or an RNA polymerase inhibitor.

### Claim 23 (depends on 22)

The method of claim 22, wherein the cytotoxin is an RNA polymerase inhibitor which is an amatoxin.

### Claim 24 (depends on 22)

The method of claim 22, wherein the cytotoxin is an antimitotic agent which is selected from the group consisting of a maytansine, an auristatin, a pyrrolobenzodiazepine (PBD), and a calicheamicin.

### Claim 25 (depends on 7)

The method of claim 7, wherein an extracellular domain of the CAR comprises an scFv antibody or a single chain T cell receptor (scTCR).

### Claim 26 (depends on 7)

The method of claim 7, wherein said tumor has a tumor antigen which is selected from the group consisting of CD19, CD22, CD30, CD7, BCMA, CD137, CD22, CD20, AFP, GPC3, MUC1, mesothelin, CD38, PD1, EG, MG7, BCMA, TAC1, CEA, PSCA, CEA, HER2, MUC1, CD33, ROR2, NKR-2, PSCA, CD28, TAA, NKG2D, and CD123.

### Claim 27 (depends on 7)

The method of claim 7, wherein a cytoplasmic domain of the CAR comprises a CD28 cytoplasmic signaling domain, a CD3 zeta cytoplasmic signaling domain, an OX40 cytoplasmic signaling domain, and/or a CD137 (4-1BB) cytoplasmic signaling domain.

### Claim 28 (depends on 7)

The method of claim 7, wherein the human subject having cancer has a cancer selected from the group consisting of leukemia, adult advanced cancer, pancreatic cancer, non-resectable pancreatic cancer, colorectal cancer, metastatic colorectal cancer, ovarian cancer, triple-negative breast cancer, hematopoietic/lymphoid cancer, colon cancer liver metastasis, small cell lung cancer, non-small cell lung cancer, B-cell lymphoma, relapsed or refractory B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large cell lymphoma, relapsed or refractory diffuse large cell lymphoma, anaplastic large cell lymphoma, primary mediastinal B-cell lymphoma, recurrent mediastinal, refractory mediastinal large B-cell lymphoma, large B-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, relapsed or refractory non-Hodgkin lymphoma, refractory aggressive non-Hodgkin lymphoma, B-cell non-Hodgkin lymphoma, refractory non-Hodgkin lymphoma, colorectal carcinoma, gastric carcinoma, pancreatic carcinoma, triple-negative invasive breast carcinoma, renal cell carcinoma, lung squamous cell carcinoma, hepatocellularcarcinoma, urothelial carcinoma, leukemia, B-cell leukemia, B-cell acute lymphocytic leukemia, B-cell acute lymphoblastic leukemia, adult acute lymphoblastic leukemia, B-cell prolymphocytic leukemia, childhood acute lymphoblastic leukemia, refractory childhood acute lymphoblastic leukemia, acute leukemia, acute lymphoblastic leukemia, acute lymphocytic leukemia, prolymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, recurrent plasma cell myeloma, refractory plasma cell myeloma, multiple myeloma, relapsed or refractory multiple myeloma, multiple myeloma of bone, malignant glioma of brain,
myelodysplastic syndrome, EGFR-positive colorectal cancer, glioblastoma multiforme, neoplasms, blastic plasmacytoid dendritic cell neoplasms, liver metastases, solid tumors, advanced solid tumors, mesothelin positive tumors, and hematological malignancies.

### Claim 29 (depends on 1)

The method of claim 1, wherein administering the immune cell expressing a CAR to said human subject occurs about 12 hours to about 21 days after step (a).

### Claim 30 (depends on 1)

The method of claim 1, wherein administering the anti-CD5 ADC to the human subject in step (a) reduces endogenous lymphocytes.