Anti-CD276 polypeptides, proteins, and chimeric antigen receptors

Inventors

Orentas, Rimas JZhu, ZhongyuMackall, Crystal LDimitrov, Dimiter SSt. Croix, BradleySaha, Saurabh

Assignees

Biomed Valley Discoveries IncUS Department of Health and Human Services

Publication Number

US-9790282-B2

Publication Date

2017-10-17

Expiration Date

2033-03-25

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Abstract

Polypeptides and proteins that specifically bind to and immunologically recognize CD276 are disclosed. Chimeric antigen receptors (CARs), anti-CD276 binding moieties, nucleic acids, recombinant expression vectors, host cells, populations of cells, and pharmaceutical compositions relating to the polypeptides and proteins are also disclosed. Methods of detecting the presence of cancer in a mammal and methods of treating or preventing cancer in a mammal are also disclosed.

Core Innovation

The invention provides polypeptides and proteins comprising antigen binding domains of anti-CD276 antibodies that specifically recognize and bind to CD276, also known as B7-H3, which is expressed or overexpressed on various human tumors including pediatric solid tumors and adult carcinomas. The inventive polypeptides and proteins may elicit antigen-specific responses against CD276, enabling them to target CD276-expressing cancer cells.

Without being bound to a specific theory, the invention believes that these polypeptides and proteins can facilitate the targeting and destruction of CD276-expressing cancer cells, reduce or eliminate cancer cells, facilitate infiltration of immune cells or effector molecules to tumor sites, and enhance or extend anti-cancer responses.

The invention also discloses chimeric antigen receptors (CARs) comprising an antigen binding domain formed by these polypeptides or proteins, a transmembrane domain, and an intracellular T cell signaling domain. Methods of detecting cancer in mammals and methods of treating or preventing cancer in mammals using these inventive materials are also provided.

Claims Coverage

The claims cover 27 inventive features primarily directed to polypeptides, proteins, chimeric antigen receptors, anti-CD276 binding moieties, conjugates, nucleic acids, expression vectors, host cells, pharmaceutical compositions, and methods related to CD276 targeting.

Polypeptides comprising specific heavy and light chains binding to CD276

Polypeptides comprising a heavy and light chain of either SEQ ID NOs: 1-6, 11-16, or 20-25 that specifically bind to CD276.

Specific polypeptide embodiments with defined sequences

Polypeptides comprising SEQ ID NOs: 7 and 8, 17 and 18, or 26 and 27.

Proteins comprising first and second polypeptide chains binding CD276

Proteins comprising a first polypeptide chain with heavy chain SEQ ID NOs: 1-3, 11-13, or 20-22 and a second polypeptide chain with light chain SEQ ID NOs: 4-6, 14-16, or 23-25 that bind CD276.

Protein embodiments comprising specific heavy and light chain sequences

Proteins comprising first polypeptide chain SEQ ID NO: 7, 17, or 26 and second polypeptide chain SEQ ID NO: 8, 18, or 27.

Polypeptides comprising linker amino acid sequences

Polypeptides further comprising a linker amino acid sequence, including linkers comprising SEQ ID NO: 115.

Polypeptides with defined combined sequences

Polypeptides comprising SEQ ID NO: 10, 19, or 28.

Chimeric antigen receptors comprising anti-CD276 antigen binding domains

Chimeric antigen receptors comprising an antigen binding domain formed by the defined polypeptides, a transmembrane domain, and an intracellular T cell signaling domain.

Transmembrane domain compositions in CARs

CARs wherein the transmembrane domain comprises CD8 and/or CD28, including sequences of SEQ ID NO: 29, 30, or 31.

Intracellular T cell signaling domains in CARs

CARs wherein the intracellular signaling domain comprises one or more of CD28 (SEQ ID NO: 32 or 35), CD137 (SEQ ID NO: 33 or 37), and CD3 zeta (SEQ ID NOs: 34, 36, and 38).

CARs comprising specific amino acid sequences

CARs comprising any one of SEQ ID NOs: 39-47 and 122-130.

Anti-CD276 binding moieties comprising specified polypeptides

Anti-CD276 binding moieties such as antibodies, Fab fragments, diabodies, single-chain variable fragments (scFv), or disulfide-stabilized variable region fragments comprising the polypeptides.

Conjugates comprising polypeptides linked to effector molecules

Conjugates comprising the polypeptides conjugated to an effector molecule, including drugs, toxins, labels, small molecules, or antibodies.

Effector molecules comprising specific toxins

Conjugates wherein the effector molecule is a Pseudomonas exotoxin variant such as PE4E, PE40, PE38, PE25, PE38QQR, PE38KDEL, PE-LR, or PE35.

Nucleic acids encoding the polypeptides

Nucleic acids comprising nucleotide sequences encoding the polypeptides such as those selected from SEQ ID NOs: 57-59.

Nucleic acids encoding CARs

Nucleic acids encoding CARs selected from SEQ ID NOs: 48-56 and 131-139.

Nucleotide sequences encoding variable region pairs

Nucleic acids comprising nucleotide sequences encoding pairs of variable regions (SEQ ID NOs: 116 and 117; 118 and 119; or 120 and 121).

Recombinant expression vectors comprising the nucleic acids

Recombinant expression vectors comprising the nucleic acid sequences encoding polypeptides or CARs.

Isolated host cells comprising the recombinant expression vectors

Isolated host cells transformed or transfected with the recombinant expression vectors.

Populations of cells comprising isolated host cells

Populations of cells comprising at least one isolated host cell with the recombinant expression vector.

Pharmaceutical compositions comprising the polypeptides

Pharmaceutical compositions containing the polypeptides and pharmaceutically acceptable carriers.

Methods of detecting cancer using polypeptides

Methods of detecting cancer in a mammal by contacting a sample with the polypeptides to form a complex and detecting the complex as indicative of cancer presence.

Use of polypeptides in treating cancer

Polypeptides expressed in host cells for use in treating cancer in a mammal.

The claims comprehensively cover polypeptides and proteins with specific anti-CD276 binding sequences, chimeric antigen receptors with defined transmembrane and signaling domains, various forms of anti-CD276 binding moieties, conjugates with effector molecules including toxins, nucleic acids encoding these molecules, recombinant vectors, host cells, pharmaceutical compositions, and methods for cancer detection and treatment.

Stated Advantages

Targets and destroys CD276-expressing cancer cells.

Reduces or eliminates cancer cells.

Facilitates infiltration of immune cells and effector molecules to tumor sites.

Enhances and extends anti-cancer immune responses.

Documented Applications

Methods of detecting the presence of cancer in a mammal by using the polypeptides, proteins, CARs, or related compositions to form detectable complexes with cancer cells.

Methods of treating or preventing cancer in mammals using the inventive polypeptides, proteins, CARs, nucleic acids, recombinant vectors, host cells, and pharmaceutical compositions.

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