Granted Patent B2

US 12,630,645 · App. 17/925,883 · Granted May 19, 2026

Inventors: Christoph Herbel (Bergisch Gladbach, DE); Dominik Eckardt (Bergisch Gladbach, DE); Vera Dittmer (Bergisch Gladbach, DE); Manuel Martinez-Osuna (Bergisch Gladbach, DE); Jutta Kollet (Bergisch Gladbach, DE); Thorsten Olaf Hardt (Bergisch Gladbach, DE); Andreas Bosio (Bergisch Gladbach, DE)

Assignee: Miltenyi Biotec B.V. & Co. KG

C07K16/30A61K40/11A61K40/31A61K40/421A61K40/4254C07K14/7051C07K16/2803A61K2239/29A61K2239/59C07K2319/02C07K2319/03C07K2319/20

Abstract

Claims (83)

1 . An in vitro combination comprising:

a) an antigen binding domain specific for CD90 and

b) an antigen binding domain specific for CD326

for use in treatment of human cancer comprising cancerous cells that co-express CD90 and CD326.

2 . The in vitro combination according to claim 1 , wherein said combination further comprises:

a) a tagged first polypeptide having the antigen binding domain specific for CD90,

b) a tagged second polypeptide having the antigen binding domain specific for CD326, wherein the tag of the first polypeptide and the tag of the second polypeptide are identical, and

c) an immune cell comprising a chimeric antigen receptor (CAR), the CAR comprising:

i) an antigen binding domain specific for said tag of the first and second polypeptides,

ii) a transmembrane domain, and

iii) an intracellular signaling domain comprising a primary signaling domain and at least one costimulatory domain.

3 . The in vitro combination of claim 2 , wherein the concentration of said tagged first polypeptide is below the activation threshold of said CAR expressed in said immune cell, wherein the concentration of said tagged second polypeptide is below the activation threshold of said CAR expressed in said immune cell, wherein the combined concentrations of the tagged first polypeptide and the tagged second polypeptide is above the activation threshold of said CAR expressed in said immune cell.

4 . The in vitro combination of claim 3 , wherein the concentration of said tagged first polypeptide is at least 10% below the concentration that is needed when the CAR is activated solely by said tagged first polypeptide, i.e. without the presence of said tagged second polypeptide, and wherein the concentration of said tagged second polypeptide is at least 10% below the concentration that is needed when the CAR is activated solely by said tagged second polypeptide, i.e. without the presence of said tagged first polypeptide.

5 . The in vitro combination according to claim 1 , wherein said combination further comprises an immune cell comprising a first CAR and a second CAR,

the first CAR comprising:

i) the first antigen binding domain specific for CD90 or CD326,

ii) a transmembrane domain, and

iii) a first intracellular signaling domain,

wherein said first intracellular signaling domain is a primary signaling domain comprising an immunoreceptor tyrosine-based activation motif (ITAM), wherein said first intracellular signaling domain does not comprise a costimulatory domain,

and the second CAR comprising:

i) the second antigen binding domain specific for CD326 if the first antigen binding domain of the first CAR is specific for CD90, or the second antigen binding domain specific for CD90 if the first antigen binding domain of the first CAR is specific for CD326,

ii) a transmembrane domain, and

iii) a second intracellular signaling domain,

wherein said second intracellular signaling domain comprises one or more co-stimulatory signaling domains,

and wherein said immune cell becomes maximally cytotoxic only when said first signaling domain and said second signaling domain are both activated by said CD90 and CD326, respectively.

6 . The in vitro combination according to claim 1 , wherein said combination further comprises an immune cell comprising a first CAR and a second CAR,

the first CAR comprising:

i) the first antigen binding domain specific for a tag of a tagged polypeptide, wherein said polypeptide has an antigen binding domain specific for CD90 or CD326,

ii) a transmembrane domain, and

iii) a first intracellular signaling domain,

wherein said first intracellular signaling domain is a primary signaling domain comprising an immunoreceptor tyrosine-based activation motif (ITAM), wherein said first intracellular signaling domain does not comprise a costimulatory domain,

and the second CAR comprising:

i) the second antigen binding domain specific for CD326 if said tagged polypeptide specifically binds to CD90, or the second antigen binding domain specific for CD90 if said tagged polypeptide specifically binds to CD326,

ii) a transmembrane domain, and

iii) a second intracellular signaling domain,

wherein said second intracellular signaling domain comprises one or more co-stimulatory signaling domains,

and wherein said immune cell becomes maximally cytotoxic only when said first signaling domain is activated by said tag and said second signaling domain is activated by said CD90 or CD326,

and wherein said combination further comprises said tagged polypeptide.

7 . The in vitro combination according to claim 1 , wherein said combination further comprises an immune cell comprising a first CAR and a second CAR,

the first CAR comprising:

i) the first antigen binding domain specific for CD90 or CD326,

ii) a transmembrane domain, and

iii) a first intracellular signaling domain,

wherein said first intracellular signaling domain is a primary signaling domain comprising an immunoreceptor tyrosine-based activation motif (ITAM), wherein said first intracellular signaling domain does not comprise a costimulatory domain,

and the second CAR comprising:

i) the second antigen binding domain specific for a tag of a tagged polypeptide wherein said polypeptide has an antigen binding domain specific for CD326 if the first antigen binding domain of the first CAR is specific for CD90, or the second antigen binding domain specific for a tagged polypeptide, wherein said polypeptide has an antigen binding domain specific for CD90 if the first antigen binding domain of the first CAR is specific for CD326,

ii) a transmembrane domain, and

iii) a second intracellular signaling domain,

wherein said second intracellular signaling domain comprises one or more co-stimulatory signaling domains,

and wherein said immune cell becomes maximally cytotoxic only when said first signaling domain is activated by said CD90 or CD326 and said second signaling domain is activated by said tag,

and wherein said combination comprises said tagged polypeptide.

8 . The in vitro combination according to claim 1 , wherein said combination further comprises an immune cell comprising a first CAR and a second CAR,

the first CAR comprising:

i) the first antigen binding domain specific for a first tag of a first tagged polypeptide, wherein said first polypeptide has an antigen binding domain specific for CD90 or CD326,

ii) a transmembrane domain, and

iii) a first intracellular signaling domain,

wherein said first intracellular signaling domain is a primary signaling domain comprising an immunoreceptor tyrosine-based activation motif (ITAM), wherein said first intracellular signaling domain does not comprise a costimulatory domain,

and the second CAR comprising:

i) the second antigen binding domain specific for a second tag of a second tagged polypeptide, wherein said second polypeptide has an antigen binding domain specific for CD326 if said first tagged polypeptide specifically binds to CD90, or the second antigen binding domain specific for a second tagged polypeptide wherein said second polypeptide has an antigen binding domain specific for CD90 if said first tagged polypeptide specifically binds to CD326,

ii) a transmembrane domain, and

iii) a second intracellular signaling domain,

wherein said second intracellular signaling domain comprises one or more co-stimulatory signaling domains,

and wherein said immune cell becomes maximally cytotoxic only when said first signaling domain and said second signaling domain are both activated by said first tag and said second tag, respectively,

and wherein said combination comprises said first tagged polypeptide and said second tagged polypeptide.

9 . The in vitro combination according to claim 1 , for use in treatment of a human ovarian cancer comprising cancerous cells that co-express CD90 and CD326.

10 . A method of treating a subject having a human cancer comprising cancerous cells that co-express CD90 and CD326, wherein the method comprises:

a) administering to said subject a tagged first polypeptide having an antigen binding domain specific for CD90, and

b) administering to said subject a tagged second polypeptide having an antigen binding domain specific for CD326, wherein the tag of the first polypeptide and the tag of the second polypeptide are identical, and

c) administering to said subject an immune cell comprising a chimeric antigen receptor (CAR), the CAR comprising:

i) an antigen binding domain specific for said tag of the first and second polypeptides,

ii) a transmembrane domain, and

iii) an intracellular signaling domain comprising a primary signaling domain and at least one costimulatory domain, thereby treating the subject having said human cancer.

11 . The method of claim 10 , wherein said human cancer is an ovarian cancer.

12 . The method of claim 10 , wherein the concentration of said tagged first polypeptide is below the activation threshold of said CAR expressed in said immune cell, wherein the concentration of said tagged second polypeptide is below the activation threshold of said CAR expressed in said immune cell, wherein the combined concentrations of the tagged first polypeptide and the tagged second polypeptide is above the activation threshold of said CAR expressed in said immune cell.

13 . The method of claim 12 , wherein the concentration of said tagged first polypeptide is at least 10% below the concentration that is needed when the CAR is activated solely by said tagged first polypeptide, i.e. without the presence of said tagged second polypeptide, and wherein the concentration of said tagged second polypeptide is at least 10% below the concentration that is needed when the CAR is activated solely by said tagged second polypeptide, i.e. without the presence of said tagged first polypeptide.

14 . The method of claim 10 , wherein said antigen binding domain specific for CD90 of the tagged polypeptide comprises SEQ ID NO: 1 and SEQ ID NO: 2, and wherein said an antigen binding domain specific for CD326 comprises SEQ ID NO: 3 and SEQ ID NO: 4.

15 . An in vitro combination comprising an antigen binding domain specific for CD90 and an antigen binding domain specific for CD326, wherein the in vitro combination comprises:

a) a tagged first polypeptide having the antigen binding domain specific for CD90;

b) a tagged second polypeptide having the antigen binding domain specific for CD326, wherein the tag of the first polypeptide and the tag of the second polypeptide are identical; and

c) an immune cell comprising a chimeric antigen receptor (CAR), the CAR comprising:

i) an antigen binding domain specific for the tag of the first polypeptide and the tag of the second polypeptide;

ii) a transmembrane domain, and

iii) an intracellular signaling domain comprising a primary signaling domain and at least one costimulatory domain.

Assignments (1)

Priority Claims (1)

Continuity (1)

References Cited (22)