WO2009032949A2 - High affinity anti-prostate stem cell antigen (psca) antibodies for cancer targeting and detection - Google Patents

High affinity anti-prostate stem cell antigen (psca) antibodies for cancer targeting and detection Download PDF

Info

Publication number
WO2009032949A2
WO2009032949A2 PCT/US2008/075291 US2008075291W WO2009032949A2 WO 2009032949 A2 WO2009032949 A2 WO 2009032949A2 US 2008075291 W US2008075291 W US 2008075291W WO 2009032949 A2 WO2009032949 A2 WO 2009032949A2
Authority
WO
WIPO (PCT)
Prior art keywords
construct
psca
antibody
minibody
cancer
Prior art date
Application number
PCT/US2008/075291
Other languages
French (fr)
Other versions
WO2009032949A3 (en
Inventor
Anna M. Wu
Robert E. Reiter
Eric J. Lepin
James D. Marks
Yu Zhou
Original Assignee
The Regents Of The University Of California
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The Regents Of The University Of California filed Critical The Regents Of The University Of California
Priority to JP2010524150A priority Critical patent/JP6126773B2/en
Priority to US12/676,348 priority patent/US8940298B2/en
Priority to EP08799192A priority patent/EP2197491A4/en
Priority to CA2698343A priority patent/CA2698343C/en
Publication of WO2009032949A2 publication Critical patent/WO2009032949A2/en
Publication of WO2009032949A3 publication Critical patent/WO2009032949A3/en
Priority to US14/562,269 priority patent/US9527919B2/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C07K16/3069—Reproductive system, e.g. ovaria, uterus, testes, prostate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
    • A61K51/1093—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody conjugates with carriers being antibodies
    • A61K51/1096—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody conjugates with carriers being antibodies radioimmunotoxins, i.e. conjugates being structurally as defined in A61K51/1093, and including a radioactive nucleus for use in radiotherapeutic applications
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/08—Drugs for disorders of the urinary system of the prostate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/10—Drugs for disorders of the urinary system of the bladder
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57434—Specifically defined cancers of prostate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A61K39/39558—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against tumor tissues, cells, antigens
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/626—Diabody or triabody
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • G01N33/57492—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds localized on the membrane of tumor or cancer cells

Definitions

  • the agent should at least demonstrate bivalent, as opposed to monovalent binding.
  • the overall size, shape, and domain composition of the agent can be varied to suit the final application.
  • Engineered fragments that exhibit optimal targeting in humans may be slightly different from formats that are optimal in mice.
  • the invention incorporates a humanized set of antibody variable regions, as well as human hinge and constant regions. Additional embodiments would include fully human variable regions.
  • the proteins can be expressed in a variety of systems, including microbial, insect cells, mammalian cell culture and transgenic animals.
  • the agent can be ricin, ricin A-chain, doxorubicin, daunorubicin, taxol, ethiduim bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxy anthracin dione, actinomycin D, diphteria toxin, Pseudomonas exotoxin (PE) A, PE40, abrin, arbrin A chain, modeccin A chain, alpha-sarcin, gelonin mitogellin, retstrictocin, phenomycin, enomycin, curicin, crotin, calicheamicin, sapaonaria officinalis inhibitor, maytansinoids, or glucocorticoidricin.
  • PE Pseudomonas exotoxin
  • FIG. 1 Schematic overview of anti-PSCA minibody.
  • a gene encoding the minibody is assembled in the order V L -lmker-V H -hinge-C H 3, with the hinge and C H 3 domains derived from human IgGl.
  • the protein self assembles into 80-kDa dimers.
  • the invention provides an antigen binding protein construct selected from the group consisting of a minibody, a diabody, scFv and scFv-Fc wherein the selected construct comprises CDR regions of an anti-PSCA antibody.
  • the binding protein construct will comprise at least one CDR region selected from a CDR-Ll, CDR-L2, CDR-L3, CDR-Hl, CDR-H2, or CDR-H3 from an anti-PSCA antibody.
  • the protein binding construct may comprise all 3 light chain CDR regions or all three heavy chain CDR regions.
  • a “minibody” is an engineered antibody construct comprised of the variable heavy (VH) and variable light (VL) chain domains of a native antibody fused to the hinge region and to the CH3 domain of the immunoglobulin molecule (see, Figure 1).
  • Minibodies are thus small versions of whole antibodies encoded in a single protein chain which retain the antigen binding region, and the CH3 domain which to permit assembly into a bivalent molecule and the antibody hinge to accommodate dimerization by disulfide linkages.
  • native antibodies are comprised of four chains, two heavy and two light. The size, valency and affinity of the minibody is particularly suited for in vivo targeting.
  • polypeptide peptide
  • protein protein
  • amino acid polymers capply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non- naturally occurring amino acid polymer.
  • amino acid sequences one of skill will recognize that individual substitutions, deletions or additions to a nucleic acid, peptide, polypeptide, or protein sequence which alters, adds or deletes a single amino acid or a small percentage of amino acids in the encoded sequence is a "conservatively modified variant" where the alteration results in the substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants are in addition to and do not exclude polymorphic variants, interspecies homologs, and alleles of the invention.
  • Antibodies exist, e.g., as intact immunoglobulins or as a number of well- characterized fragments produced by digestion with various peptidases.
  • pepsin digests an antibody below the disulfide linkages in the hinge region to produce F(ab)' 2; a dimer of Fab which itself is a light chain joined to V H -C H I by a disulfide bond.
  • the F(ab)' 2 may be reduced under mild conditions to break the disulfide linkage in the hinge region, thereby converting the F(ab)' 2 dimer into an Fab' monomer.
  • phage display technology can be used to identify antibodies and heteromeric Fab fragments that specifically bind to selected antigens (see, e.g., McCafferty et al, Nature 348:552-554 (1990); Marks et al, Biotechnology 10:779-783 (1992)).
  • Antibodies can also be made bispecific, i.e., able to recognize two different antigens (see, e.g., WO 93/08829, Traunecker et al, EMBO J. 10:3655-3659 (1991); and Suresh et al, Methods in Enzymology 121:210 (1986)).
  • Antibodies can also be heteroconjugates, e.g., two covalently joined antibodies, or immunotoxins (see, e.g., U.S. Patent No. 4,676,980 , WO 91/00360; WO 92/200373; and EP 03089).
  • a humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues are often referred to as import residues, which are typically taken from an import variable domain. Humanization can be essentially performed following the method of Winter and co-workers (see, e.g., Jones et al., Nature 321:522-525 (1986); Riechmann et al, Nature 332:323-327 (1988); Verhoeyen et al, Science 239:1534-1536 (1988) and Presta, Curr. Op. Struct. Biol.
  • the antibody is conjugated to an "effector" moiety.
  • the effector moiety can be any number of molecules, including labeling moieties such as radioactive labels or fluorescent labels, or can be a therapeutic moiety.
  • the antibody modulates the activity of the protein.
  • effector moieties include, but are not limited to, an anti-tumor drug, a toxin, a radioactive agent, a cytokine, a second antibody or an enzyme.
  • the invention provides an embodiment wherein the antibody of the invention is linked to an enzyme that converts a prodrug into a cytotoxic agent.
  • the immunoconjugate can be used for targeting the effector moiety to a PSCA- positive cell, particularly cells, which overexpress the PCSA protein. Such differences can be readily apparent when viewing the bands of gels with approximately similarly loaded with test and controls samples..
  • the invention provides antigen binding protein constructs used to systemically to treat cancer (e.g., prostate, pancreatic or bladder cancer) alone or when conjugated with an effector moiety.
  • PSCA-targeting constructs conjugated with toxic agents, such as ricin, as well as unconjugated antibodies can be useful therapeutic agents naturally targeted to PSCA bearing cancer cells.
  • Such constructs can be useful in blocking invasiveness
  • the antigen-binding protein constructs of the invention can be used to treat cancer.
  • the construct is joined to at least a functionally active portion of a second protein or toxic molecule having therapeutic activity.
  • the second protein can include, but is not limited to, an enzyme, lymphokine, oncostatin or toxin.
  • Suitable toxins include doxorubicin, daunorubicin, taxol, ethiduim bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxy anthracin dione, actinomycin D, diphteria toxin, Pseudomonas exotoxin (PE) A, PE40, ricin, abrin, glucocorticoid and radioisotopes.
  • PE Pseudomonas exotoxin
  • the specified antibodies bind to a particular protein at least two times the background and more typically more than 10 to 100 times background.
  • Specific binding to an antibody under such conditions requires a construct be selected for its specificity for a particular protein.
  • a variety of immunoassay formats may be used to select constructs specifically immunoreactive with PSCA.
  • solid-phase ELISA immunoassays are routinely used to select antibodies specifically immunoreactive with a protein ⁇ see, e.g., Harlow & Lane, Using Antibodies, A Laboratory Manual (1998) for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity) .
  • pharmaceutically acceptable salts or “pharmaceutically acceptable carrier” is meant to include salts of the active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein.
  • base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent.
  • pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt.
  • acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent.
  • pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like.
  • the neutral forms of the compounds may be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner.
  • the parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but otherwise the salts are equivalent to the parent form of the compound for the purposes of the present invention.
  • Dosages will depend upon various factors generally appreciated by those of skill in the art, including without limitation the type of cancer and the severity, grade, or stage of the cancer, the binding affinity and half life of the agents used, the desired steady-state antibody concentration level, frequency of treatment, and the influence of chemotherapeutic agents used in combination with the treatment method of the invention.
  • Typical daily doses may range from about 0.1 to 100 mg/kg. Doses in the range of 10-500 mg of the constructs or their immunoconjugates per week may be effective and well tolerated, although even higher weekly doses may be appropriate and/or well tolerated.
  • the principal determining factor in defining the appropriate dose is the amount of a particular agent necessary to be therapeutically effective in a particular context. Repeated administrations may be required in order to achieve tumor inhibition or regression.
  • Initial loading doses may be higher. The initial loading dose may be administered as an infusion. Periodic maintenance doses may be administered similarly, provided the initial dose is well tolerated.
  • the agents may be injected directly into the bladder.
  • compositions for administration will commonly comprise an agent as described herein dissolved in a pharmaceutically acceptable carrier, preferably an aqueous carrier.
  • a pharmaceutically acceptable carrier preferably an aqueous carrier.
  • aqueous carriers can be used, e.g., buffered saline and the like. These solutions are sterile and generally free of undesirable matter.
  • These compositions may be sterilized by conventional, well known sterilization techniques.
  • the compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents, toxicity adjusting agents and the like, for example, sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like.
  • concentration of active agent in these formulations can vary widely, and will be selected primarily based on fluid volumes, viscosities, body weight and the like in accordance with the particular mode of administration selected and the patient's needs.
  • the formulation may also provide additional active compounds, including, chemotherapeutic agents, cytotoxic agents, cytokines, growth inhibitory agent, and anti- hormonal agent.
  • the active ingredients may also prepared as sustained-release preparations (e.g., semi-permeable matrices of solid hydrophobic polymers (e.g., polyesters, hydrogels (for example, poly (2-hydroxyethyl-methacrylate), or poly (vinylalcohol) ), polylactides.
  • the methods of the invention may be practiced together with other cancer therapies (e.g, radical prostatectomy), radiation therapy (external beam or brachytherapy), hormone therapy (e.g., orchiectomy, LHRH- analog therapy to suppress testosterone production, anti-androgen therapy), or chemotherapy.
  • Radical prostatectomy involves removal of the entire prostate gland plus some surrounding tissue. This treatment is used commonly when the cancer is thought not to have spread beyond the tissue. Radiation therapy is commonly used to treat prostate cancer that is still confined to the prostate gland, or has spread to nearby tissue. If the disease is more advanced, radiation may be used to reduce the size of the tumor.
  • Hormone therapy is often used for patients whose prostate cancer has spread beyond the prostate or has recurred.
  • Preferred pharmaceutical preparations deliver one or more constructs according to the invention, optionally in combination with one or more chemotherapeutic agents or immunotherapeutic agents, in a sustained release formulation.
  • the construct may be administered therapeutically as a sensitizing agent that increases the susceptibility of tumor cells to other cytotoxic cancer therapies, including chemotherapy, radiation therapy, immunotherapy and hormonal therapy.
  • Oligonucleotides and vectors used in the construction of the library include; Gap 5': 5'-TTAAGCTTCTGCAGGCTAGTG-S' (SEQ ID NO:2); Gap 3': 5'- GAGACCGAGGAGAGGGTTAGG-S 1 (SEQ ID NO:3); pYD2 inside the Ncol-Notl restriction sites (Razai A, et al. J MoI Biol. 2005; 35 1 : 1 58).
  • Example 4 This example describes the biochemical characterization of anti-PSCA minibodies.
  • Flow Cytometry was conducted to assess cellular PSCA binding activity.
  • An EBV transformed B-cell lymphoma cell line stably transfected with PSCA were used. Briefly, cells 5 x 10' were incubated for 30 min on ice with 100 ⁇ l of minibody at 2ug/ml concentration. Cells were washed and stained with goat anti-hFc PE conjugated antibody at 1:100 dilution.
  • mice were anesthetized using 2% isoflurane, positioned in a prone position along the long axis of the microPET scanner and imaged. Acquisition time was 10 minutes (1 bed position), and images were reconstructed using a filtered backprojection reconstruction algorithm. Images were displayed and regions of interest (ROI) were drawn as described in figure 6 and quantified using AMIDE (Loening and Gambhir, Molecular Imaging 2: 13 1,2003). After scanning, tumors, liver, spleen, kidney, lung, and blood were excised, weighed and counted in a well counter (Cobra II AutoGamma, Packard, IL). Background, crossover, and decay corrections were done. Results were calculated as percentage of injected dose per gram of tissue (% ID/g).

Abstract

The present invention provides novel high affinity antibodies and fragments thereof that bind to the cancer antigen PSCA. The antibodies of the present invention may be used for cancer diagnosis, prognosis, treatment, visualization, and the like. The present invention also provides methods for the detection, visualization, and treatment of various cancers expressing PSCA.

Description

HIGH AFFINITYANTI-PROSTATE STEM CELL ANTIGEN (PSCA) ANTIBODIES FOR CANCER TARGETING AND DETECTION
CROSS-REFERENCES TO RELATED APPLICATIONS [0001] NOT APPLICABLE
STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT [0002] This invention was made with Government support of Grant No. C A092131, awarded by the NIH/NCI. The Government has certain rights in this invention.
REFERENCE TO A "SEQUENCE LISTING," A TABLE, OR A COMPUTER
PROGRAM LISTING APPENDIX SUBMITTED ON A COMPACT DISK. [0003] NOT APPLICABLE
BACKGROUND OF THE INVENTION
[0004] Prostate stem cell antigen (PSCA) a cell surface glycoprotein expressed in normal human prostate and bladder is over-expressed in prostate cancers (40% of primary tumors and 60-100% of lymph node and bone marrow metastases). It is also highly expressed in transitional carcinomas of the bladder and pancreatic carcinoma. 1G8, an anti-PSCA mouse monoclonal antibody specific for PSCA demonstrated anti-tumor targeting activity in vivo (Gu Z, et al. Cancer Res. 2005; 65:9495). This antibody was humanized by grafting on a human framework (Trastuzumab) and named 2B3 (Olafsen T, et al. J. Immunotherapy 2007; 30:396).
[0005] The invention addresses the need for agents that have appropriate pharmacodynamic properties to target and image tumors that express PSCA. There is a tremendous need in the field for effective agents to image cancers with sensitivity and specificity, particularly early stage tumors or ones with early metastasis not imageable by traditional means. As PSCA is highly expressed by most prostate, bladder and pancreatic tumors, it is an important target in the detection, diagnosis, prognosis, and treatment of these cancers. The current invention describes an innovative constructs with optimal characteristics for tumor imaging and targeting. They may also be used for tumor targeting of gene therapy, radioactivity therapy, and may have therapeutic utility by themselves.
BRIEF SUMMARY OF THE INVENTION
[0006] This invention describes engineered antibodies that recognize a novel cell surface marker in prostate and other cancers with high affinity. These genetically engineered antibody fragments are tailored specifically for in vivo use for targeting and detection.
[0007] The invention addresses the need for agents that have appropriate pharmacodynamic properties to target and image tumors that express PSCA. There is a tremendous need in the field for effective agents to image cancers with sensitivity and specificity, particularly early stage tumors or ones with early metastasis not imageable by traditional means. PSCA is highly expressed by most prostate, bladder and pancreatic tumors and is a promising target. The current invention describes an innovative molecule with optimal characteristics for tumor imaging. It may also be useful for tumor targeting of gene therapy, radioactivity or may have therapeutic utility by itself.
[0008] There are multiple embodiments. For instance, there are a variety of engineered antibody formats, such as scFvs, diabodies, triabodies, minibodies, and scFv-Fc. In general, the agent should at least demonstrate bivalent, as opposed to monovalent binding. The overall size, shape, and domain composition of the agent can be varied to suit the final application. Engineered fragments that exhibit optimal targeting in humans may be slightly different from formats that are optimal in mice. Since one goal is human application, the invention incorporates a humanized set of antibody variable regions, as well as human hinge and constant regions. Additional embodiments would include fully human variable regions. The proteins can be expressed in a variety of systems, including microbial, insect cells, mammalian cell culture and transgenic animals.
[0009] For imaging purposes, a variety of radionuclides can be attached to the engineered antibodies for detection with gamma or SPECT cameras, or PET scanners. For therapy one can attach drugs, toxins, cytokines, enzymes, or other therapeutic moieties for PSCA-targeted delivery to tumors. The engineered PSCA-specific antibodies can be coupled to nanosensors for detection (in vitro or in vivo) or nanoparticles for delivery (in vivo). One can also incorporate the PSCA antibody fragments into viral vectors for targeted gene therapy of tumors. [0010] The invention addresses the unmet need for imaging of cancer, in early diagnosis or diagnosis of metastatic disease. In particular, there is a critical need for better agents for imaging prostate cancer for detection and staging. PSCA antibody fragment imaging will be very useful for imaging bone metastases and assessing response to treatment; there are no good imaging approaches currently available. Detection of pancreatic cancer is a critical need, and an imaging agent would be useful in high-risk patients. The invention describes high-affinity, highly specific engineered antibodies tailored for in vivo targeting and detection of PSCA in prostate cancer, bladder cancer, and pancreatic cancer patients.
[0011] Accordingly, in a first aspect, the invention provides high affinity PSCA antigen binding protein constructs which can be used in the treatment and detection of cancers which overexpress PSCA. In some embodiments, these constructs are minibodies, diabodies, triabodies, ScFv, or ScFv-Fc as described further below. In one embodiment, the invention provides an antigen binding protein construct directed toward a mammalian PSCA protein (e.g., human, murine) selected from the group consisting of a minibody, a diabody, and scFv- Fc wherein the selected construct has VL and VH domains that are substantially identical, respectively, to the VL domain and the VH domain of an anti-PSCA antibody. For example, the construct can be a minibody in which the VL and VH chain variable domains of the anti- PSCA antibody are fused to part of the hinge region of an antibody, an amino acid linker and the CH3 domain of an immunoglobulin molecule. In other embodiments, the construct is a diabody.
[0012] In embodiments, where the construct is a minibody or diabody or scFV-Fc, the anti- PSCA antibody can be a humanized antibody and the CH3 domain is from a human immunoglobulin molecule. In preferred embodiments, the anti-PSCA antibody is 2B3 or 1G8. In yet other embodiments, the construct is a minibody having VH and VLdomains substantially identical to an scFv fragment designated herein as Al 1, A2, or C5. In still further embodiments, the construct has a CH3 domain is from a human immunoglobulin molecule. For instance, the construct may have a hinge region and CH3 domain are the human IgG hinge region and human CH3 domain.
[0013] In some embodiments, the anti-PSCA antibody is a monoclonal antibody designated 1G8 (ATCC No. HB-12612), 2A2 (ATCC No. HB-1203), 2H9 (ATCC No. HB-12614), 3C5 (ATCC No. HB-12616), 3E6 (ATCC No. HB12618), 3G3 (ATCC No. HB-12615), or 4A10 (ATCC No. HB- 12617). [0014] The constructs according to the invention can also be linked to therapeutic agents or detectable markers. In some embodiments, the therapeutic agent is a cytotoxic agent. For instance, the agent can be ricin, ricin A-chain, doxorubicin, daunorubicin, taxol, ethiduim bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxy anthracin dione, actinomycin D, diphteria toxin, Pseudomonas exotoxin (PE) A, PE40, abrin, arbrin A chain, modeccin A chain, alpha-sarcin, gelonin mitogellin, retstrictocin, phenomycin, enomycin, curicin, crotin, calicheamicin, sapaonaria officinalis inhibitor, maytansinoids, or glucocorticoidricin. In other embodiments, the therapeutic agent is a radioactive isotope. The radioactive isotope can be selected, for instance, from the group consisting Of212Bi, 1311, 111In, 9OY and 186Re. In other embodiments the construct is linked to an anti-cancer pro-drug activating enzyme capable of converting a pro-drug to its active form.
[0015] In another preferred embodiments of the above, the PSCA targeted by the anti- PSCA antibody is human PSCA.
[0016] hi another aspect, the invention provides an antigen binding protein construct selected from the group consisting of a minibody, a diabody, scFv and scFv-Fc wherein the selected construct has VL and VH domains that are substantially identical to the VL and VH domains of 2B3 or an scFv variant designated herein as Al 1, A2, or C5. In other embodiments, the binding constructs of the present invention may comprise one or more mutations found in the variant antibodies Al 1, A2, or C5.
[0017] hi yet another aspect, the present invention provides an antigen binding protein construct selected from the group consisting of a minibody, a diabody, scFv and scFv-Fc wherein the selected construct comprises CDR regions of an anti-PSCA antibody, hi certain embodiments, the anti-PSCA antibody will bind to PSCA with an affinity equal to or greater than the antibody designated 2B3. In other embodiments, the anti-PSCA antibody may be an affinity matured antibody, wherein the affinity matured antibody comprises a higher affinity for PSCA than does the parental antibody.
[0018] In another aspect, the anti-PSCA construct according to the invention is labeled with a detectable marker. The marker can be for instance, a radioisotope, a fluorescent compound, a bioluminescent compound, chemiluminescent compound, a metal chelator or an enzyme. Many radionuclides may be used as imaging labels, including without limitation, 1241, 86Y, 18F, 94Tc, and the like. One of skill in the art will know of other radionuclides particularly well suited for use in the present invention. [0019] In further other embodiments of any of the above, the invention provides a pharmaceutical composition of the constructs according to the invention.
[0020] In another aspect still, the invention provides methods for treating a subject having cancer (e.g, prostate, pancreatic or bladder cancer), or inhibiting the growth of a prostate cancer cell expressing a Prostate Stem Cell Antigen (PSCA) protein comprising contacting the cancer cell (e.g., prostate, bladder, pancreatic cancer cell, with a construct according to the invention in an amount effective to inhibit the growth of the cancer cell. The method can kill the cancer cell, hi some embodiments, the construct recognizes and binds the PSCA protein as shown below beginning with leucine at amino acid position 22 and ending with alanine at amino acid position 99. In additional embodiments, the method further comprises administering to a chemotherapeutic drug, radiation therapy, hi some embodiments, the subject is also treated with hormone ablation therapy or hormone antagonist therapy.
[0021] The treatments may be given to the patient or subject by intravenously, intraperitoneally, intramuscularly, intratumorally, or intradermally. In some embodiments, the contacting comprises administering the construct directly into a prostate cancer, a bladder cancer, a pancreatic cancer or a metastasis thereof.
[0022] In another aspect, the invention provides methods of detecting a cancerous cell in a subject by contacting the cancer cell with a construct which bears a detectable marker. The methods can be used in screening patients at increased risk of cancer or to monitory response to therapy or to develop a prognosis for the cancer (e.g., prostate, bladder, or pancreatic cancers. The methods are particularly advantageous in detecting metastases of the cancer.
BRIEF DESCRIPTION OF THE DRAWINGS
[0023] Figure 1: Schematic overview of anti-PSCA minibody. A gene encoding the minibody is assembled in the order VL-lmker-VH-hinge-CH3, with the hinge and CH3 domains derived from human IgGl. The protein self assembles into 80-kDa dimers.
[0024] Figure 2: Comparison of 2B3 ScFv variant protein sequences. The VL sequence and part of the linker are in the top frame, the rest of the linker and VH sequences are in the lower frame. Also indicated are the CDRs: underlined wild type sequence. The mutations are highlighted in different shading. The parental ScFv P-2B3 sequence (SEQ ID NO:4) and variant C5 (SEQ ID NO:5), A2 (SEQ ID NO:6), and Al 1 (SEQ ED NO:7) ScFv sequences are shown. [0025] Figure 3: DNA sequence (SEQ ID NO: 8) and translated protein sequence of the parental 2B3 minibody (SEQ ID NO:9). Also indicated are the starting points of the following protein segments: signal peptide (for mammalian secretion), light chain variable region (VL), the 15 amino acid interdomain peptide linker, heavy chain variable region (VH), human IgGl hinge sequence and 10 amino acid extension, and the human IgGl CHj domain followed by two stop codons.
[0026] Figure 4: SDS-PAGE analysis of the four minibodies in non-reducing conditions: parental (p), Al 1, A2 and C5. The arrow indicates the expected molecular weight of a minibody.
[0027] Figure 5: Size-exclusion HPLC of Al 1 minibody showing homogenous peak at expected molecular size.
[0028] Figure 6: Affinity ranking. (A) by Competitive ELISA binding assay: Plates were coated with CEA and biotinylated intact, chimeric anti-CEA antibody was used as probe. (B) by flow cytometry: 5pg/ml of each minibody was incubated with PSCA expressing cells. Cells were then stained with anti Human Fc PE conjugated.
[0029] Figure 7: Co-registered microPET/microCT scan of a nude mouse bearing LAPC- 9AD (PSCA-positive human prostate cancer) xenografts. The mouse was injected with 1-1 24 radiolabeled Al 1 minibody variant and scanned serially. A, B, C; saggital sections; D, E, F; coronal sections. A, D; coregistered microPET and microCT, showing ROI (region of interest) as a white rectangle. B, E; microPET images and ROI C, F; microPET images only.
[0030] Figure 8: Biodistribution and microPET Imaging ranking of the 2B3 minibody variants. Biodistribution unit is a %ID/g values of weighed tissues in y- counter after 21 or 25 hours injection time. MicroPET Imaging values were obtained on the average values of 4 ROIs as shown in figure 6 and described in the materials and methods.
[0031] Figure 9: Summary of PSCA data and properties.
[0032] Figure 10: Depictions of various constructs according to the invention.
[0033] Figure 11: Schema for producing 2B3 variants.
[0034] Figure 12: MicroPET imaging and biodistribution data for various constructs.
[0035] Figure 13: MicroPET imaging and biodistribution results for All and parent 2B3. [0036] Figure 14: MicroPET imaging and biodistribution data for pancreatic cancer Capan-1 xenographic mice using parental 2B3 and variant Al 1 anti-PSCA minibodies.
[0037] Figure 15: MicroPET imaging and biodistribution data for pancreatic cancer MIA PaCa-2 xenographic mice using parental 2B3 and variant Al 1 anti-PSCA minibodies.
[0038] Figure 16: Amino acid sequences of parental 2B3 and variant A2, Al 1, and C5 anti-PSCA minibodies.
DETAILED DESCRIPTION OF THE INVENTION
[0039] Prolonged clearance kinetics have hampered the development of intact antibodies as imaging and therapeutic agents, despite their ability to effectively deliver radionuclides to tumor targets in vivo. Here, we also report genetically engineered antibody fragments which display rapid, high-level tumor uptake coupled with rapid clearance from the circulation in a nude mouse model to allow ready detection of tumors in vivo
[0040] The 2B3 antibody was humanized by grafting on a human framework (Trastuzumab) and named (Olafsen T, et al. J. Immunotherapy 2007; 30:396; which is incorporated by reference in its entirety with respect to the description, biological activity, and making of the antibody). Antibody fragments of 2B3 have been generated for PET imaging application. One of these antibody fragments, the 2B3 minibody has demonstrated rapid and specific localization to PSCA-expressing tumors in a murine model. However, humanization has decreased the affinity of the intact antibody for PSCA from a Kd of 2.6 nM to 16.7 n M. hi addition, reformatting of intact antibody into antibody fragments can also affect the binding efficiency. It has been shown that quantitative tumor retention of ScFvs increases with affinity but only to a threshold close to 10"9 M (Adams GP, et al. Cancer Res. 2001;61:4750). There are no published data on the effect of minibody affinity on tumor targeting and imaging properties. Here, we describe the generation of three high affinity anti- PSCA ScFvs, and the subsequent generation and characterization of three high affinity anti- PSCA minibodies. We generated minibody fragments with better tumor targeting/imaging aptitude, hi some embodiments, with the regard to the constructs of the invention, there is a proviso that the construct comprises a VH or VL domain that is not identical to a corresponding domain or the 2B3 antibody.
[0041] In one embodiment, the present invention provides antigen binding constructs selected from the group consisting of an antibody, a minibody, a diabody, an scFv, an scFv- Fc, and the like, wherein the VL and VH domains are substantially identical to those found in 2B3. In a second embodiment, the antigen binding construct may comprise one or more mutations found in an antibody variant designated herein as Al 1, A2, or C5. hi a third embodiment, the binding construct comprises at least one mutation at a residue corresponding to an amino acid of SEQ ID NO:4 selected from the group consisting of T5, SlO, V15, S91, S123, S131, N179, T182, 1194, A203, G213, Q228, and a combination thereof. In certain embodiments, the at least one mutation comprises a mutation corresponding to a mutation in SEQ ID NO:4 selected from the group comprising T5I, SlOI, V15M, S91G, Δ123, S131Y, N179Y, T182S, I194M, A203V, E213K, Q228R, and a combination thereof. In other embodiments, the binding constructs of the invention comprise mutations corresponding to those found in variants Al l, A2, or C5.
[0042] In a certain embodiment, the present invention provides a minibody wherein the VL and VH domains are substantially identical to those found in SEQ ID NO: 10. In one embodiment, a minibody of the present invention comprises an amino acid sequence of SEQ ID NO: 10. In other embodiments, the minibody comprises an amino acid sequence that is substantially identical to SEQ ID NO: 10, wherein said minibody binds to PSCA with a higher affinity than a minibody of SEQ ID NO: 10. In yet other embodiments, the minibody may comprise one or more mutations at a residue corresponding to an amino acid of SEQ ID NO:10 selected from the group comprising T5, SlO, V15, S91, S123, S131, N179, T182, 1194, A203, G213, Q228, and a combination thereof, hi yet another embodiment, the construct may comprise a sequence selected from SEQ ID NOS: 11, 12, and 13.
[0043] In one embodiment, the invention provides an antigen binding protein construct selected from the group consisting of a minibody, a diabody, scFv and scFv-Fc wherein the selected construct comprises CDR regions of an anti-PSCA antibody. Li one embodiment, the binding protein construct will comprise at least one CDR region selected from a CDR-Ll, CDR-L2, CDR-L3, CDR-Hl, CDR-H2, or CDR-H3 from an anti-PSCA antibody. In yet other embodiments, the protein binding construct may comprise all 3 light chain CDR regions or all three heavy chain CDR regions. In one embodiment, the protein binding constructs of the present invention may comprise all of the CDR regions of an anti-PSCA antibody, hi certain embodiments, the anti-PSCA antibody will bind to PSCA with an affinity equal to or greater than the antibody designated 2B3. In other embodiments, the anti-PSCA antibody may be an affinity matured antibody, wherein the affinity matured antibody comprises a higher affinity for PSCA than does the parental antibody. In particular embodiments, the parental anti-PSCA antibody may be selected from the group consisting of 1G8 (ATCC No. HB-12612), 2A2 (ATCC No. HB-1203), 2H9 (ATCC No. HB-12614), 3C5 (ATCC No. HB- 12616), 3E6 (ATCC No. HB12618), 3G3 (ATCC No. HB-12615), 4A10 (ATCC No. HB- 12617), and 2B3. In other embodiments, the CDRs may be selected from those found in SEQ ID NOS:10, 11, 12, and 13.
[0044] A "minibody" is an engineered antibody construct comprised of the variable heavy (VH) and variable light (VL) chain domains of a native antibody fused to the hinge region and to the CH3 domain of the immunoglobulin molecule (see, Figure 1). Minibodies are thus small versions of whole antibodies encoded in a single protein chain which retain the antigen binding region, and the CH3 domain which to permit assembly into a bivalent molecule and the antibody hinge to accommodate dimerization by disulfide linkages. In contrast, native antibodies are comprised of four chains, two heavy and two light. The size, valency and affinity of the minibody is particularly suited for in vivo targeting. Expression in bacterial or mammalian cells is simplified because minibodies can be produced as single amino acid chains (see, U.S. Patent No. 5,837,821) which is incorporated by reference herein in its entirety and particularly with reference to minibodies, their structure, ways of making them, and their suitable pharmaceutical formulations.
[0045] A 'diabody" comprises a first polypeptide chain which comprises a heavy (VH) chain variable domain connected to a light chain variable domain (VL) on the first polypeptide chain (VH-VL) connected by a peptide linker that is too short to allow pairing between the two domains on the first polypeptide chain and a second polypeptide chain comprising a light chain variable domain (VL) linked to a heavy chain variable domain VH on the second polypeptide chain (VL-VH) connected by a peptide linker that is too short to allow pairing between the two domains on the second polypeptide chain. The short linkages force chain pairing between the complementary domains of the first and the second polypeptide chains and promotes the assembly of a dimeric molecule with two functional antigen binding sites.
[0046] To construct bispecific diabodies the V-domains of different antibodies (e.g., antibody A and antibody B) are fused to create the two chains (e.g., VHA-VLB, VHB-VLA). Each chain is inactive in binding to antigen, but recreates the functional antigen binding sites of antibodies A and B on pairing with the other chain. [0047] PSCA and its expression in cancer of the prostate, bladder, and pancreas is disclosed in U.S. Patent No. 6,756,036 which is incorporated by reference in its entirety. The human PSCA translated amino acid sequence is:
MKAVLLALLMAGLALQPGTALLCVSCKAQVSNEDCLQV
ENCTQLGEQCWTARIRAVGLLTVISKGCSLNCVDOS QDYYVGKKHITCCDTDLCNASGAHALQPAAAILALLPAL 6LLLWGPGOL
(SEQ ID NO:1)
[0048] The terms "substantially identical" in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., about 80% identity, preferably at least 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity) over the referenced sequences or portions, when compared and aligned for maximum correspondence over a comparison window or designated region) as measured using a BLAST or BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection {see, e.g., NCBI web site http://www.ncbi.nlm.nih.gov/BLAST/ or the like). The definition also includes sequences that have deletions and/or additions, as well as those that have substitutions. Preferably, the sequence identity is at least 85%, 90%, 95% 97% between two referenced domains. In some embodiments, the difference in sequence is just by one, two, three or four, or from five to 12, amino acids as to referenced sequence or domain. As described below, the preferred algorithms can account for gaps and the like. Preferably, identity exists over a region that is at least about 15 amino acids or nucleotides in length, or more preferably over a region that is 15-50 amino acids or nucleotides in length. In other embodiments, the identity may exist over a region that is at least about 50, 100, 150, 200, or more amino acids.
[0049] For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Preferably, default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters.
[0050] A "comparison window", as used herein, includes reference to a segment of any one of the number of contiguous positions selected over a specified range of residues (e.g., 20 to 50, usually about 50 to about 200, more usually about 100 to about 150) in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Methods of alignment of sequences for comparison are well-known in the art. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J MoI. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat 'I. Acad. ScL USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by manual alignment and visual inspection (see, e.g., Current Protocols in Molecular Biology (Ausubel et al., eds. 1995 supplement)).
[0051] A preferred example of algorithm that is suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al, Nuc. Acids Res. 25:3389-3402 (1977) and Altschul et al, J. MoI Biol. 215:403-410 (1990), respectively. BLAST and BLAST 2.0 are used, with the parameters described herein, to determine percent sequence identity for the nucleic acids and proteins of the invention. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov/). This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive- valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always > 0) and N (penalty score for mismatching residues; always < 0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, M=5, N=-4 and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength of 3, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sd. USA 89:10915 (1989)) alignments (B) of 50, expectation (E) of 10, M=5, N=-4, and a comparison of both strands.
[0052] The terms "polypeptide," "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms capply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non- naturally occurring amino acid polymer.
[0053] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise, may be referred to by their commonly accepted single-letter codes.
[0054] As to amino acid sequences, one of skill will recognize that individual substitutions, deletions or additions to a nucleic acid, peptide, polypeptide, or protein sequence which alters, adds or deletes a single amino acid or a small percentage of amino acids in the encoded sequence is a "conservatively modified variant" where the alteration results in the substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants are in addition to and do not exclude polymorphic variants, interspecies homologs, and alleles of the invention.
[0055] The following eight groups each contain amino acids that are conservative substitutions for one another: 1) Alanine (A), Glycine (G); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W); 7) Serine (S), Threonine (T); and 8) Cysteine (C), Methionine (M) (see, e.g., Creighton, Proteins (1984)).
[0056] A "label" or a "detectable moiety'Or "detectable marker" is a composition detectable by spectroscopic, photochemical, biochemical, immunochemical, chemical, or other physical means. For example, useful labels include 32P, fluorescent dyes, electron-dense reagents, enzymes (e.g., as commonly used in an ELISA), biotin, digoxigenin, or haptens and proteins which can be made detectable, e.g., by incorporating a radiolabel into the peptide or used to detect antibodies specifically reactive with the peptide.
[0057] "Antibody" refers to a polypeptide comprising a framework region from an immunoglobulin gene or fragments thereof that specifically binds and recognizes an antigen. The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon, and mu constant region genes, as well as the myriad immunoglobulin variable region genes. Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively. Typically, the antigen-binding region of an antibody will be most critical in specificity and affinity of binding.
[0058] An exemplary immunoglobulin (antibody) structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" (about 25 kD) and one "heavy" chain (about 50-70 kD). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The terms variable light chain (VL) and variable heavy chain (VH) refer to these light and heavy chains respectively.
[0059] Antibodies exist, e.g., as intact immunoglobulins or as a number of well- characterized fragments produced by digestion with various peptidases. Thus, for example, pepsin digests an antibody below the disulfide linkages in the hinge region to produce F(ab)'2; a dimer of Fab which itself is a light chain joined to VH-CHI by a disulfide bond. The F(ab)'2 may be reduced under mild conditions to break the disulfide linkage in the hinge region, thereby converting the F(ab)'2 dimer into an Fab' monomer. The Fab' monomer is essentially Fab with part of the hinge region (see Fundamental Immunology (Paul ed., 3d ed. 1993). While various antibody fragments are defined in terms of the digestion of an intact antibody, one of skill will appreciate that such fragments may be synthesized de novo either chemically or by using recombinant DNA methodology. Thus, the term antibody, as used herein, also includes antibody fragments either produced by the modification of whole antibodies, or those synthesized de novo using recombinant DNA methodologies (e.g., single chain Fv) or those identified using phage display libraries (see, e.g., McCafferty et al, Nature 348:552-554 (1990))
[0060] For preparation of suitable antibodies or constructs of the invention and for use according to the invention, e.g., recombinant, monoclonal, or polyclonal antibodies, many techniques known in the art can be used (see, U.S. Patent Application Publication No. 20070196274 and U.S. Patent Application Publication No. 20050163782, which are each incorporated by reference in their entirities, particularly with respect to minibody and diabody design) (see, e.g., Kohler & Milstein, Nature 256:495-497 (1975); Kozbor et al, Immunology Today 4: 72 (1983); Cole et al, pp. 77-96 in Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc. (1985); Coligan, Current Protocols in Immunology (1991); Harlow & Lane, Antibodies, A Laboratory Manual (1988); and Goding, Monoclonal Antibodies: Principles and Practice (2d ed. 1986)). The genes encoding the heavy and light chains of an antibody of interest can be cloned from a cell, e.g., the genes encoding a monoclonal antibody can be cloned from a hybridoma and used to produce a recombinant monoclonal antibody. Gene libraries encoding heavy and light chains of monoclonal antibodies can also be made from hybridoma or plasma cells. Random combinations of the heavy and light chain gene products generate a large pool of antibodies with different antigenic specificity (see, e.g., Kuby, Immunology (3r ed. 1997)). Techniques for the production of single chain antibodies or recombinant antibodies (U.S. Patent 4,946,778, U.S. Patent No. 4,816,567) can be adapted to produce antibodies to polypeptides of this invention. Also, transgenic mice, or other organisms such as other mammals, may be used to express humanized or human antibodies (see, e.g., U.S. Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, Marks et al, Bio/Technology 10:779-783 (1992); Lonberg et al, Nature 368:856-859 (1994); Morrison, Nature 368:812-13 (1994); Fishwild et al, Nature Biotechnology 14:845-51 (1996); Neuberger, Nature Biotechnology 14:826 (1996); and Lonberg & Huszar, Intern. Rev. Immunol. 13:65-93 (1995)). Alternatively, phage display technology can be used to identify antibodies and heteromeric Fab fragments that specifically bind to selected antigens (see, e.g., McCafferty et al, Nature 348:552-554 (1990); Marks et al, Biotechnology 10:779-783 (1992)). Antibodies can also be made bispecific, i.e., able to recognize two different antigens (see, e.g., WO 93/08829, Traunecker et al, EMBO J. 10:3655-3659 (1991); and Suresh et al, Methods in Enzymology 121:210 (1986)). Antibodies can also be heteroconjugates, e.g., two covalently joined antibodies, or immunotoxins (see, e.g., U.S. Patent No. 4,676,980 , WO 91/00360; WO 92/200373; and EP 03089).
[0061] The antibodies of the invention include derivatives that are modified, i.e., by the covalent attachment of any type of molecule to the antibody. For example, the antibody derivatives include, without limitation, antibodies that have been modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein, and the like. Any of numerous chemical modifications may be carried out by known techniques, including, but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. Additionally, the derivative may contain one or more non-natural amino acids.
[0062] Methods for humanizing or primatizing non-human antibodies are well known in the art. Generally, a humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues are often referred to as import residues, which are typically taken from an import variable domain. Humanization can be essentially performed following the method of Winter and co-workers (see, e.g., Jones et al., Nature 321:522-525 (1986); Riechmann et al, Nature 332:323-327 (1988); Verhoeyen et al, Science 239:1534-1536 (1988) and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Accordingly, such humanized antibodies are chimeric antibodies (U.S. Patent No. 4,816,567), wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.
[0063] A "chimeric antibody" is an antibody molecule in which (a) the constant region, or a portion thereof, is altered, replaced or exchanged so that the antigen binding site (variable region) is linked to a constant region of a different or altered class, effector function and/or species, or an entirely different molecule which confers new properties to the chimeric antibody, e.g., an enzyme, toxin, hormone, growth factor, drug, etc.; or (b) the variable region, or a portion thereof, is altered, replaced or exchanged with a variable region having a different or altered antigen specificity. The preferred antibodies of, and for use according to the invention include humanized and/or chimeric monoclonal antibodies.
[0064] Li some embodiments, the antibody is conjugated to an "effector" moiety. The effector moiety can be any number of molecules, including labeling moieties such as radioactive labels or fluorescent labels, or can be a therapeutic moiety. In one aspect the antibody modulates the activity of the protein. Such effector moieties include, but are not limited to, an anti-tumor drug, a toxin, a radioactive agent, a cytokine, a second antibody or an enzyme. Further, the invention provides an embodiment wherein the antibody of the invention is linked to an enzyme that converts a prodrug into a cytotoxic agent. [0065] The immunoconjugate can be used for targeting the effector moiety to a PSCA- positive cell, particularly cells, which overexpress the PCSA protein. Such differences can be readily apparent when viewing the bands of gels with approximately similarly loaded with test and controls samples.. Examples of cytotoxic agents include, but are not limited to ricin, doxorubicin, daunorubicin, taxol, ethiduim bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxy anthracin dione, actinomycin D, diphteria toxin, Pseudomonas exotoxin (PE) A, PE40, abrin, and glucocorticoid and other chemotherapeutic agents, as well as radioisotopes. Suitable detectable markers include, but are not limited to, a radioisotope, a fluorescent compound, a bioluminescent compound, chemiluminescent compound, a metal chelator or an enzyme.
[0066] In some embodiments, the invention provides antigen binding protein constructs used to systemically to treat cancer (e.g., prostate, pancreatic or bladder cancer) alone or when conjugated with an effector moiety. PSCA-targeting constructs conjugated with toxic agents, such as ricin, as well as unconjugated antibodies can be useful therapeutic agents naturally targeted to PSCA bearing cancer cells. Such constructs can be useful in blocking invasiveness
[0067] Additionally, the antigen-binding protein constructs of the invention can be used to treat cancer. In such a situation, the construct is joined to at least a functionally active portion of a second protein or toxic molecule having therapeutic activity. The second protein can include, but is not limited to, an enzyme, lymphokine, oncostatin or toxin. Suitable toxins include doxorubicin, daunorubicin, taxol, ethiduim bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxy anthracin dione, actinomycin D, diphteria toxin, Pseudomonas exotoxin (PE) A, PE40, ricin, abrin, glucocorticoid and radioisotopes.
[0068] Techniques for conjugating therapeutic agents to constructs according to the invention are well known (see, e.g., Arnon et al., "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy", in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., "Antibodies For Drug Delivery"in Controlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); Thorpe, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review" in Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985); and Thorpe et al., "The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates", Immunol. Rev., 62:119-58 (1982)).
[0069] The phrase "specifically (or selectively) binds" to an antibody or "specifically (or selectively) immunoreactive with," when referring to a protein or peptide or construct according to the invention, refers to a binding reaction that is determinative of the presence of the protein, often in a heterogeneous population of proteins and other biologies. Thus, under designated immunoassay conditions, the specified antibodies bind to a particular protein at least two times the background and more typically more than 10 to 100 times background. Specific binding to an antibody under such conditions requires a construct be selected for its specificity for a particular protein. A variety of immunoassay formats may be used to select constructs specifically immunoreactive with PSCA. For example, solid-phase ELISA immunoassays are routinely used to select antibodies specifically immunoreactive with a protein {see, e.g., Harlow & Lane, Using Antibodies, A Laboratory Manual (1998) for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity) .
[0070] By "therapeutically effective dose or amount" herein is meant a dose that produces effects for which it is administered. The exact dose and formulation will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques {see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Remington: The Science and Practice of Pharmacy, 20th Edition, Gennaro, Editor (2003), and Pickar, Dosage Calculations (1999)). [0071] The term "pharmaceutically acceptable salts" or "pharmaceutically acceptable carrier" is meant to include salts of the active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. When compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, e.g., Berge et ah, Journal of Pharmaceutical Science 66:1-19 (1977)). Certain specific compounds of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts. Other pharmaceutically acceptable carriers known to those of skill in the art are suitable for the present invention.
[0072] The neutral forms of the compounds may be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but otherwise the salts are equivalent to the parent form of the compound for the purposes of the present invention.
[0073] The methods find particular application in the diagnosis, prognosis and treatment of cancers which overexpress PSCA, for example, prostate, pancreatic and bladder cancers. In certain embodiments the methods are applied to hormone refractory or therapy resistant cancers. In certain embodiments the methods are applied to metastatic cancers. [0074] Treatment will generally involve the repeated administration of the constructs and their immunoconjugates via an acceptable route of administration such as intravenous injection (IV), at an effective dose. Dosages will depend upon various factors generally appreciated by those of skill in the art, including without limitation the type of cancer and the severity, grade, or stage of the cancer, the binding affinity and half life of the agents used, the desired steady-state antibody concentration level, frequency of treatment, and the influence of chemotherapeutic agents used in combination with the treatment method of the invention. Typical daily doses may range from about 0.1 to 100 mg/kg. Doses in the range of 10-500 mg of the constructs or their immunoconjugates per week may be effective and well tolerated, although even higher weekly doses may be appropriate and/or well tolerated. The principal determining factor in defining the appropriate dose is the amount of a particular agent necessary to be therapeutically effective in a particular context. Repeated administrations may be required in order to achieve tumor inhibition or regression. Initial loading doses may be higher. The initial loading dose may be administered as an infusion. Periodic maintenance doses may be administered similarly, provided the initial dose is well tolerated.
[0075] Direct administration of the constructs is also possible and may have advantages in certain contexts. For example, for the treatment of bladder carcinoma, the agents may be injected directly into the bladder.
[0076] In another embodiment, the invention provides polynucleotides comprising a nucleotide sequence encoding an antibody of the invention and fragments thereof. In one embodiment, the present invention provides an expression vector encoding for an antibody or fragment thereof of the present invention. In another embodiment, the present invention provides polynucleotides encoding an antibody of the present invention for use in gene therapy or in vivo administration.
[0077] "Nucleic acid" refers to deoxyribonucleotides or ribonucleotides and polymers thereof in either single- or double-stranded form, and complements thereof. The term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides. Examples of such analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs).
[0078] Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions) and complementary sequences, as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et ah, J. Biol. Chem. 260:2605-2608 (1985); Rossolini et al., Mol. Cell. Probes 8:91-98 (1994)). The term nucleic acid is used interchangeably with gene, cDNA, mRNA, oligonucleotide, and polynucleotide.
Methods of Administration and Formulation
[0079] The constructs are administered to a subject in accord with known methods, such as intravenous administration, e.g., as a bolus or by continuous infusion over a period of time, by intramuscular, intraperitoneal, intracerobrospinal, subcutaneous, intra-articular, intrasynovial, intrathecal, oral, topical, or inhalation routes. Intravenous or subcutaneous administration is preferred. The administration may be local or systemic.
[0080] The compositions for administration will commonly comprise an agent as described herein dissolved in a pharmaceutically acceptable carrier, preferably an aqueous carrier. A variety of aqueous carriers can be used, e.g., buffered saline and the like. These solutions are sterile and generally free of undesirable matter. These compositions may be sterilized by conventional, well known sterilization techniques. The compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents, toxicity adjusting agents and the like, for example, sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like. The concentration of active agent in these formulations can vary widely, and will be selected primarily based on fluid volumes, viscosities, body weight and the like in accordance with the particular mode of administration selected and the patient's needs.
[0081] Thus, a typical pharmaceutical composition for intravenous administration will vary according to the agent. Actual methods for preparing parenterally administrable compositions will be known or apparent to those skilled in the art and are described in more detail in such publications as Remington's Pharmaceutical Science, 15th ed., Mack Publishing Company, Easton, Pa. (1980).
[0189] The pharmaceutical compositions can be administered in a variety of unit dosage forms depending upon the method of administration. For example, unit dosage forms suitable for oral administration include, but are not limited to, powder, tablets, pills, capsules and lozenges. It is recognized that constructs when administered orally, should be protected from digestion. This is typically accomplished either by complexing the molecules with a composition to render them resistant to acidic and enzymatic hydrolysis, or by packaging the molecules in an appropriately resistant carrier, such as a liposome or a protection barrier. Means of protecting agents from digestion are well known in the art.
[0082] Pharmaceutical formulations, particularly, constructs and immunoconjugates and inhibitors for use with the present invention can be prepared by mixing a construct having the desired degree of purity with optional pharmaceutically acceptable carriers, excipients or stabilizers. Such formulations can be lyophilized formulations or aqueous solutions. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations used. Acceptable carriers, excipients or stabilizers can be acetate, phosphate, citrate, and other organic acids; antioxidants (e.g., ascorbic acid) preservatives low molecular weight polypeptides; proteins, such as serum albumin or gelatin, or hydrophilic polymers such as polyvinylpyllolidone; and amino acids, monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents; and ionic and non-ionic surfactants (e.g., polysorbate); salt-forming counter-ions such as sodium; metal complexes (e. g. Zn-protein complexes); and/or non-ionic surfactants. The construct can be formulated at a concentration of between 0.5 - 200 mg/ml, or between 10-50 mg/ml.
[0083] The formulation may also provide additional active compounds, including, chemotherapeutic agents, cytotoxic agents, cytokines, growth inhibitory agent, and anti- hormonal agent. The active ingredients may also prepared as sustained-release preparations (e.g., semi-permeable matrices of solid hydrophobic polymers (e.g., polyesters, hydrogels (for example, poly (2-hydroxyethyl-methacrylate), or poly (vinylalcohol) ), polylactides. The antibodies and immunocongugates may also be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin microcapsules and poly- (methylmethacylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions.
[0084] The compositions can be administered for therapeutic or prophylactic treatments, hi therapeutic applications, compositions are administered to a patient suffering from a disease (e.g., cancer) in a "therapeutically effective dose." Amounts effective for this use will depend upon the severity of the disease and the general state of the patient's health. Single or multiple administrations of the compositions may be administered depending on the dosage and frequency as required and tolerated by the patient. A "patient" or "subject" for the purposes of the present invention includes both humans and other animals, particularly mammals. Thus the methods are applicable to both human therapy and veterinary applications. In the preferred embodiment the patient is a mammal, preferably a primate, and in the most preferred embodiment the patient is human. Other known cancer therapies can be used in combination with the methods of the invention. For example, the compositions for use according to the invention may also be used to target or sensitize a cell to other cancer therapeutic agents such as 5FU, vinblastine, actinomycin D, cisplatin, methotrexate, and the like.
[0085] In other embodiments, the methods of the invention may be practiced together with other cancer therapies (e.g, radical prostatectomy), radiation therapy (external beam or brachytherapy), hormone therapy (e.g., orchiectomy, LHRH- analog therapy to suppress testosterone production, anti-androgen therapy), or chemotherapy. Radical prostatectomy involves removal of the entire prostate gland plus some surrounding tissue. This treatment is used commonly when the cancer is thought not to have spread beyond the tissue. Radiation therapy is commonly used to treat prostate cancer that is still confined to the prostate gland, or has spread to nearby tissue. If the disease is more advanced, radiation may be used to reduce the size of the tumor. Hormone therapy is often used for patients whose prostate cancer has spread beyond the prostate or has recurred. The objective of hormone therapy is to lower levels of the male hormones, androgens and thereby cause the prostate cancer to shrink or grow more slowly. Luteinizing hormone-releasing hormone (LHRH) agonists decrease the production of testosterone. These agents may be injected either monthly or longer. Two such analogs are leuprolide and goserelin. Anti-androgens (e.g., flutamide, bicalutamide, and nilutamide) may also be used. Total androgen blockade refers to the use of anti-androgens in combination with orchiectomy or LHRH analogs, the s combination is called. Chemotherapy is an option for patients whose prostate cancer has spread outside of the prostate gland and for whom hormone therapy has failed. It is not expected to destroy all of the cancer cells, but it may slow tumor growth and reduce pain. Some of the chemotherapy drugs used in treating prostate cancer that has returned or continued to grow and spread after treatment with hormonal therapy include doxorubicin (Adriamycin), estramustine, etoposide, mitoxantrone, vinblastine, and paclitaxel. Two or more drugs are often given together to reduce the likelihood of the cancer cells becoming resistant to chemotherapy. Small cell carcinoma is a rare type of prostate cancer that is more likely to respond to chemotherapy than to hormonal therapy.
[0086] The combined administrations contemplates co-administration, using separate formulations or a single pharmaceutical formulation, and consecutive administration in either order, wherein preferably there is a time period while both (or all) active agents simultaneously exert their biological activities.
[0087] The compound of choice, alone or in combination with other suitable components, can be made into aerosol formulations (i.e., they can be "nebulized") to be administered via inhalation. Aerosol formulations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen, and the like.
[0088] Formulations suitable for parenteral administration, such as, for example, by intraarticular (in the joints), intravenous, intramuscular, intratumoral, intradermal, intraperitoneal, and subcutaneous routes, include aqueous and non-aqueous, isotonic sterile injection solutions, which can contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. In the practice of this invention, compositions can be administered, for example, by intravenous infusion, orally, topically, intraperitoneally, intravesically or intrathecally. Parenteral administration, oral administration, and intravenous administration are the preferred methods of administration. The formulations of compounds can be presented in unit-dose or multi-dose sealed containers, such as ampules and vials.
[0089] Injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the kind previously described. Cells transduced by nucleic acids for ex vivo therapy can also be administered intravenously or parenterally as described above.
[0090] The pharmaceutical preparation is preferably in unit dosage form. In such form the preparation is subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form. The composition can, if desired, also contain other compatible therapeutic agents.
[0091] Preferred pharmaceutical preparations deliver one or more constructs according to the invention, optionally in combination with one or more chemotherapeutic agents or immunotherapeutic agents, in a sustained release formulation. The construct may be administered therapeutically as a sensitizing agent that increases the susceptibility of tumor cells to other cytotoxic cancer therapies, including chemotherapy, radiation therapy, immunotherapy and hormonal therapy.
[0092] In therapeutic use for the treatment of cancer, the constructs utilized in the pharmaceutical method of the invention are administered at the initial dosage of about 0.001 mg/kg to about 1000 mg/kg daily. A daily dose range of about 0.01 mg/kg to about 500 mg/kg, or about 0.1 mg/kg to about 200 mg/kg, or about 1 mg/kg to about 100 mg/kg, or about 10 mg/kg to about 50 mg/kg, can be used. The dosages, however, may be varied depending upon the requirements of the patient, the severity of the condition being treated, and the compound being employed. For example, dosages can be empirically determined considering the type and stage of cancer diagnosed in a particular patient. The dose administered to a patient, in the context of the present invention should be sufficient to effect a beneficial therapeutic response in the patient over time. Determination of the proper dosage for a particular situation is within the skill of the practitioner. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day, if desired.
[0093] The pharmaceutical preparations for use according to the invention are typically delivered to a mammal, including humans and non-human mammals. Non-human mammals treated using the present methods include domesticated animals (i.e., canine, feline, murine, rodentia, and lagomorpha) and agricultural animals (bovine, equine, ovine, porcine).
Methods of Tumor Imaging [0094] In certain embodiments, the present invention provides methods of imaging cancer cells or tumors in vivo through administration of antibodies of the invention. In one embodiment, the present invention provides a method of imaging a cancer cell in vivo, the method comprising administering a labeled anti-PSCA antibody to a mammal and imaging the antibody in vivo. The methods of the present invention may be used to image a cancer cell in mammal, including without limitation, a mouse, rat, hamster, rabbit, pig, human, and the like.
[0095] Methods of in vivo imaging are well known in the art and include without limitation, magnetic resonance imaging (MRI), nuclear magnetic resonance (NMR) (R. Weissleder, 1999, Radiology 212:609-14), computerized axial tomography (CAT) scan, cooled charged coupled device (CCD) camera optical imaging (Honigman, et al., 2001 MoI. Ther. 4:239- 249), bioluminescent optical imaging (P R Contag, et al., 1998 Nat. Med. 4:245-247), position emission tomography (PET) (M E Phelps, 1991 Neurochemical Research 16:929- 994; J G Tjuvajev, et al., 1998 Cancer Res 58:4333-4341), single photon emission computed tomography (J G. Tjuvajev, et al., 1996 Cancer Res. 45:4087-4095), microPET (reviewed in McVeigh, 2006, Circ. Res. 98:879-86), and the like.
Examples
[0096] The following examples are offered to illustrate, but not limit, the claimed invention.
[0097] Example 1
[0098] This example describes the construction of a mutant scFv yeast display library and the selection of ScFv mutants with improved PSCA binding affinity.
[0099] Oligonucleotides and vectors used in the construction of the library include; Gap 5': 5'-TTAAGCTTCTGCAGGCTAGTG-S' (SEQ ID NO:2); Gap 3': 5'- GAGACCGAGGAGAGGGTTAGG-S1 (SEQ ID NO:3); pYD2 inside the Ncol-Notl restriction sites (Razai A, et al. J MoI Biol. 2005; 35 1 : 1 58).
[0100] 2B3 ScFv gene was first cloned into the yeast display vector pYD2 (Razai A, et al. J MoI. Biol. 2005; 351:158) using Ncol-Notl restriction sites. A bacterial clone with the correct sequence was amplified and DNA extracted with QIAprep Spin Miniprep. Random mutations were introduced into the 2B3 Scfv gene using error prone PCR as follows: 2B3 ScFv gene was subject to 20 cycles of PCR with Taq in the presence of 10OmM MnC 12 to generate random mutations. The PCR product was run on an agarose gel and purified using QIAquick gel extraction. The purified PCR product was re-amplified using a proofreading DNA polymerase for 35 cycles. Both PCRs were carried out with the Gap5' and Gap3' primers. The amplified 2B3 Fv gene was again run on an agarose gel and purified using QIAquick gel extraction. Mutated scFv genes and Ncol-Notl digested pYD2 were used to transform LiAc -treated EBY 100 cells by gap repair. The resulting gene repertoire was cloned into pYD2 using gap repair to create a library of 5.9 X 105 transformants Transformation mixes were cultured and sub-cultured in SD-CAA. Library size was determined by plating serial dilutions of the transformation mixture on SD-CAA plates.
[0101] For selection, scFv display was induced by culturing in SG-CAA media plus zeocin for 24 hours at 2O0C. For the first round of selection 20 million yeast (more than 30 times the library size) were washed and resuspended in FACS buffer (phosphate-buffered saline (pH 7.4), 0.5% bovine serum albumin) to which 200 nM of PSCA human Gamma 1 fusion protein was added and incubated for 1 hour at room temperature. The concentrations of PSCA human Gamma 1 used for round 2, 3 and 4 of sorting were 5 nM, 2 nM and InM respectively. Cells were incubated for 30 minutes with secondary antibodies at 40C, washed once with FACS buffer, resuspended in 200-500 μl of FACS buffer and sorted on a FACS Aria. Typically 1% of the PSCA binding population was gated for collection. Collected cells were grown in SD- CAA media and used for the next round of sorting after induction in SG-CAA. Twenty yeast clones from the fourth round of sorting were analyzed by flow cytometry. Eight of these clones showing strong staining were selected (A2, A4, A8, A9, Al 1, A12, B5 and C5) and their DNA sequenced. A2, A4, A8, A9, A12, B5 protein sequences were identical with 10 mutations, Al 1 had 6 mutations and C5 had 4 mutations. Protein sequence comparisons of the parental 2B3 ScFv with A2, Al 1 and C5 are shown in Figure 2.
[0102] Example 2
[0103] This example describes the reformatting of mutant 2B3 scFv's into minibodies.
[0104] The parental 2B3 minibody pEE12 construct (Figure 2) was used as a backbone to generate the three 2B3 minibody affinity variants where the wild type ScFv insert was replaced by each of the three 2B3 ScFv affinity variants. The parental 2B3 minibody construct is presented in Figure 1. Briefly, VL and VH regions were fused with a 15 residue long Gly-Ser rich linker in the VL-VH orientation. This ScFv is flanked by a signal peptide upstream and the human IgGlCπ3 domain via the human IgGl hinge including a 10 residue GlySer peptide linker downstream. The final product was cloned into the PEE 12 vector of expression (Lonza Biologies, Slough, UK). This vector contains the hCMV promoter and the glutamine synthetase gene for selection (Bebbington et al., Biotechnology (N Y). 1992; 10: 169). The parental 2B3 minibody pEE12 construct was used as a backbone to generate the three 2B3 minibody affinity variants. The parental pEE12 DNA was digested with Xba I and Xho I restriction sites to remove the parental 2B3 ScFv insert that was replaced by each of the three 2B3 ScFv affinity variants. Xba I- Xho I restriction sites were added at the extremities of 2B3 ScFv affinity variants for sub-cloning in pEE12, by extension PCR.
[0105] Example 3
[0106] This example describes the expression, selection, and purification of anti-PSCA minibodies.
[0107] A total of 2 x 106 NSO mouse myeloma cells were transfected with lOug of linearized (cut with SaIT) vector DNA by electroporation and selected in glutamine-deficient media as described (Yazaki PJ, et al. J Immunol Methods. 2001 ;253: 195). Clones were screened for expression by ELISA, whereby the desired protein was captured by goat anti- human IgG (Fc specific) and detected by alkaline phosphastase (AP)-conjugated goat anti- human IgG (Fc specific) (both from Jackson ImmunoResearch Labs, West Grove,PA). The highest producing clones were expanded and brought to terminal culture.
[0108] Soluble minibodies were purified from cell culture supernatants by Protein L chromatography using a Thermal Separations Products HPLC with an in-line UV monitor, equipped with a preparative Poros 50 A column (Applied Biosystems, Foster City, CA) and analyzed on SDS-PAGE (Figure 3). The four minibodies showed similar results. All the minibodies migrated as molecular weight species of -95 kDa under non-reducing conditions and all showed good purity. In addition, the Al 1 minibody eluted at 29.5 minutes as expected when run on a calibrated size exclusion column (Figure 4).. Supernatants were loaded onto a 10x50 mm column and eluted using 0.1 M glycine pH 2.5, the pH was immediately neutralized with 2M Tris-HCl pH 8. The purified proteins were then dialyzed against PBS using a molecular porous membrane tubing (mwco: 30,000) and concentrated with a Vivascience Vivaspin 20 (mwco: 30,000). Final protein concentrations were determined by measuring UV absorbance at 280 nm, using the parental murine antibody as the standard.
[0109] Example 4 [0110] This example describes the biochemical characterization of anti-PSCA minibodies.
[0111] Size and composition: Purified proteins were analyzed by SDS-PAGE (Figure 3) under non-reducing conditions. Native structural size was determined by size exclusion columns (Superdex 75) (Pharmacia).
[0112] The ranking of the four minibodies was determined by competition ELISA and flow cytometry (figure 5). The relative affinity as measured by competition ELISA indicated that all three affinity variants had higher affinity than the parental, with an improvement of 4.4x, 3.Ox and 1.9x for A2, Al 1 and C5 respectively compared to the parental. Flow cytometry data also resulted in ranking the four minibodies in the same order when targeting PSCA expressed at the cell surface. In conclusion, the affinity ranking of the four minibodies was: A2>A1 l>C5>parental.
[0113] Competition ELISA: PSCA relative binding affinity for the minibodies was determined by competition ELISA in which microtiter plate wells were coated with purified PSCA-Fc (Olafsen T, et al. J. Immunotherapy 2007: 30:396).
[0114] Flow Cytometry : was conducted to assess cellular PSCA binding activity. An EBV transformed B-cell lymphoma cell line stably transfected with PSCA were used. Briefly, cells 5 x 10' were incubated for 30 min on ice with 100 μl of minibody at 2ug/ml concentration. Cells were washed and stained with goat anti-hFc PE conjugated antibody at 1:100 dilution.
[0115] Radioiodination: Purified minibodies were radioiodinated with the positron emitting isotope 124I( sodium iodide in 0.02 M NaOH; radionuclide purity >99%) provided by Advanced Nuclide Technologies, Indianapolis, IN as previously described (Kenanova, Olafsen et al., Cancer Res. 65:622,2005). rmmunoreactivity was assayed by incubating radioiodinated-minibody with an excess amount of SKW-PSCA+ cells for an hour at room temperature, centrifugating the cells, and counting radioactivity present into the supernatant compared to the control.
[0116] Example 5
[0117] This example describes MicroPET imaging and biodistribution studies of anti- PSCA minibodies.
[0118] All animal studies were conducted under protocols approved by the Chancellor's Animal Research Committee at the University of California, Los Angeles. Xenografts were established in 7- to 8-week-old male nude mice (Charles River Laboratories, Wilmington, MA) by s.c. inoculation of 2 XlO6 LAPCP AD cells in the shoulder region. After 14 days, when tumor masses were in the range of 100 to 300 mg, 100 μCi of isotope 124I(30-50 μg protein) was injected into the tail vein of each animal. Mice were imaged using a P4 microPET scanner (Concorde Microsystems, Inc., Knoxville, TN). To enable imaging, mice were anesthetized using 2% isoflurane, positioned in a prone position along the long axis of the microPET scanner and imaged. Acquisition time was 10 minutes (1 bed position), and images were reconstructed using a filtered backprojection reconstruction algorithm. Images were displayed and regions of interest (ROI) were drawn as described in figure 6 and quantified using AMIDE (Loening and Gambhir, Molecular Imaging 2: 13 1,2003). After scanning, tumors, liver, spleen, kidney, lung, and blood were excised, weighed and counted in a well counter (Cobra II AutoGamma, Packard, IL). Background, crossover, and decay corrections were done. Results were calculated as percentage of injected dose per gram of tissue (% ID/g).
[0119] To evaluate tumor targeting and microPET imaging efficiency, 124I-labeled minibodies were injected into Nude mice bearing LAPC-9AD tumors on the right shoulder. Wholebody micoPET and CT scans were performed at 2 Ih and/or 25h, after which the animals were sacrificed, and activity in various tissues quantified using a gamma counter. To quantify microPET imaging, four 3-dimentional ROIs were drawn in the tumors and four other ROI in soft tissues around the tumor as presented in figure 6. The ROI position and size were based on CT image information. Both biodistribution and imaging quantification are presented as a ratio of tumor signal to background. Al l minibody gave the best biodistribution and imaging data in an experiment that compared the three affinity variant (Figure 7A). Thus the affinity ranking (Figure 5) and in vivo tumor targeting/imaging ranking (Figure 7A) of the three affinity variant minibodies are different, suggesting that in our model in vivo tumor targeting/imaging effectiveness is not solely dependant on the inherent affinity of the tracer to its target. A2 which has the best affinity to PSCA did not give the best in vivo tumor targeting/imaging results. One possible explanation for this discordance is that A2 has a substitution of an asparagine into a tyrosine in VL CDR2, and that iodination of this new tyrosine could affect the binding to PSCA. In a second experiment Al 1 minibody was compared to the parental minibody for their in vivo tumor targeting/imaging effectiveness. Al l showed a 20% increase in tumor targeting (n=3) and a 14 1 % increase in microPET tumor imaging (n=2) (figure 7B). [0120] Example 6
[0121] This example describes the imaging of various pancreactic cancer tumors using anti- PSCA minibodies.
[0122] In order to evaluate the targeting and imaging potential of affinity matured anti- PSCA minibodies, 124I-labeled minibodies (parental 2B3 and variant Al 1) were injected into athymic nude mice bearing tumors that express low levels of target PSCA antigen. Briefly, xenographic mice bearing either human Capan-1 (Figure 14) or human MIA PaCa-2 (Figure 15) pancreactic tumors were injected with either 200 or 300 μg of labeled anti-PSCA minibody. Wholebody micoPET and CT scans were performed as before, and tissue radioactivity was determined. Similarly, quantification of microPET imaging and ROI position and size were determined as in example 5.
[0123] As can be seen in Figures 14 and 15, variant Al 1 anti-PSCA microbodies consistently demonstrated improved tumor to muscle specificity ratios as compared to parental 2B3 minibodies. The nearly 2-fold enhancement in in vivo specificity suggests that these variant minibodies are better suited for use in therapeutic targeting and tumor imaging than existing anti-PSCA antibodies. Notebly, tumor uptake in Capan-1 and HPAF-11 (data not shown) was roughly 2% and less than 1% in MIA PaCa-2 tumors, suggesting specific uptake.
[0124] Related art:
[0125] 1) Sundaresan, G., Yazaki, PJ., Shively, J.E., Finn, R.D., Larson, S.M., Raubitschek, A.A., Williams, L.E., Chatziioannou, A.F., Gambhir, S. S., and Wu, A.M. (2003) Iodine- 1 24 labeled engineered anti-CEA minibodies and diabodies allow highcontrast, antigen-specific small-animal PET imaging of xenografts in athymic mice. J Nucl. Med., 44:1962-1969.
[0126] 2) Olafsen, T., Gu, Z., Sherman, M.A., Leyton, J. V., Witkosky, M.E., Shively, J.E.., Raubitschek, A.A., Morrison, S.L., Wu, A.M. and Reiter, R.E. (2007) Targeting, imaging, and therapy using a humanized anti-prostate stem cell antigen (PSCA) antibody. J. Immunotherapy 30:396-405.
[0127] 3) Leyton, J.V., Olafsen, T., Sherman, M.A., Reiter, R.E., and Wu, A.M. Anti- prostate stem cell antigen (PSCA) antibody fragments for PET imaging of prostate cancer (abstract). Cancer Biotherapy & Radiopharmaceuticals 21 :391, 2006. [0128] It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes to the extent consistent with the present disclosure.

Claims

WHAT IS CLAIMED IS:
1. An antigen binding protein construct selected from the group consisting of a minibody, a diabody, scFv and scFv-Fc wherein the selected construct has VL and VH domains that are substantially identical to the VL and VH domains of a minibody or scFv designated herein as Al 1, A2, or C5.
2. An antigen binding protein construct selected from the group consisting of a minibody, a diabody, and a scFv-Fc, wherein the selected construct has VL and VH domains that are substantially identical, respectively, to the VL domain and the VH domain ofan anti-PSC A antibody.
3. The antigen binding protein construct of claim 2, wherein said anti- PSCA antibody is an affinity matured antibody.
4. The construct of any one of claims 1 to 3, wherein the construct is a minibody in which the VL and VH chain variable domains of the anti-PSCA antibody are fused to the hinge region of an antibody, an amino acid linker and the CH3 domain of an immunoglobulin molecule.
5. The construct of any one of claims 1 to 3, wherein the construct is a diabody.
6. The construct of any one of claims 1 to 5, wherein the anti-PSCA antibody is a humanized antibody and the CH3 domain is from a human immunoglobulin molecule.
7. The construct of claim 6, wherein the hinge region is a human IgG hinge region.
8. The construct of any one of claims 2 to 7, wherein the anti-PSCA antibody is 2B3.
9. The construct of claim 2, wherein the antigen binding protein is a minibody comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 10, wherein said minibody binds to PSCA with an affinity equal to or greater than that of antibody 2B3.
10. The construct of claim 9, wherein said minibody binds to PSCA with an affinity greater than that of antibody 2B3.
11. The minibody of claim 9, wherein said minibody comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 10, 11, 12, and 13.
12. The construct of claim 2 or 3, wherein the anti-PSCA antibody is a monoclonal antibody designated 1G8 (ATCC No. HB-12612), 2A2 (ATCC No. HB-1203), 2H9 (ATCC No. HB-12614), 3C5 (ATCC No. HB-12616), 3E6 (ATCC No. HB12618), 3G3 (ATCC No. HB-12615), or 4A10 (ATCC No. HB-12617).
13. An antigen binding protein construct selected from the group consisting of a minibody, a diabody, and a scFv-Fc, wherein the selected construct comprises CDR regions of an anti-PSCA antibody.
14. The construct of claim 13, wherein said anti-PSCA antibody binds to PSCA with an affinity equal to or greater than that of antibody 2B3.
15. The construct of claim 14, wherein said anti-PSCA antibody is selected from those designated 2B3, A2, Al 1, or C5.
16. The construct of any one of claims 13 to 15, wherein said construct is a minibody.
17. The construct of any one of claims 1 to 16, wherein the construct is conjugated to a therapeutic agent.
18. The construct of claim 17, wherein the therapeutic agent is a cytotoxic agent.
19. The construct of claim 18, wherein the cytotoxic agent is selected from a group consisting of ricin, ricin A-chain, doxorubicin, daunorubicin, taxol, ethiduim bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxy anthracin dione, actinomycin D, diphteria toxin, Pseudomonas exotoxin (PE) A, PE40, abrin, arbrin A chain, modeccin A chain, alpha-sarcin, gelonin mitogellin, retstrictocin, phenomycin, enomycin, curicin, crotin, calicheamicin, sapaonaria officinalis inhibitor, maytansinoids, and glucocorticoidricin.
20. The construct of claim 18, wherein the therapeutic agent is a radioactive isotope.
21. The construct of claim 20, wherein the radioactive isotope is selected from the group consisting Of212Bi, 1311, 111In, 90Y and 186Re.
22. The construct of any one of claims 1 to 16, wherein the construct is linked to an anti-cancer pro-drug activating enzyme capable of converting a pro-drug to its active form.
23. The construct of claim 2, wherein the PSCA is human PSCA.
24. The construct of any one of claims 1 to 16, wherein the construct is labeled with a detectable marker.
25. The construct according to claim 24, wherein the marker is a radioisotope, a fluorescent compound, a bioluminescent compound, chemiluminescent compound, a metal chelator or an enzyme.
26. The construct of claim 25, wherein the marker is 124I.
27. A polynucleotide encoding an anti-PSCA antibody of any one of claims 1 to 25.
28. A method for inhibiting the growth of a prostate cancer cell expressing a Prostate Stem Cell Antigen (PSCA) protein comprising contacting the cancer cell with a construct of any one of claims 1 to 23, in an amount effective to inhibit the growth of the cancer cell.
29. A method for killing a prostate cancer cell by the method of claim 28.
30. The method of claim 28, wherein the construct recognizes and binds the PSCA protein as shown in SEQ ID NO:2 beginning with leucine at amino acid position 22 and ending with alanine at amino acid position 99.
31. The method of claim 28, further comprising administering to the cell a chemo therapeutic drug.
32. The method of claim 28 further comprising administering radiation therapy to the cell.
33. A method for treating a patient suffering from a prostate, bladder, or pancreatic cancer by the method of any one of claims 28 to 32.
34. The method of claim 33, comprising administering to the patient hormone ablation therapy or hormone antagonist therapy.
35. The method of claim 34, wherein the contacting comprises administering the construct to the patient intravenously, intraperitoneally, intramuscularly, intratumorally, or intradermally.
36. The method of claim 35, wherein the contacting comprises administering the construct directly into a prostate cancer, a bladder cancer, a pancreatic cancer or a metastasis thereof.
37. A method of detecting a cancerous cell in a subject, said method comprising contacting a construct of any one of claims 24 to 26.
38. A pharmaceutical composition comprising a construct of any one of claims 2 to 1, and pharmaceutically acceptable excipient, carrier, or stabilizer.
39. The composition of claim 38, wherein the composition is a lyophilized formulations or aqueous solution.
40. A construct of any one of claims 1 to 23, for use in treating or detecting a prostate, bladder, or pancreatic cancer in a subject.
41. A method of imaging a cancer cell in vivo, comprising the steps of:
(a) administering to a mammal suspected of having cancer an antibody of any one of claims 24 to 26; and (b) detecting said antibody with a method selected from the group consisting of magnetic resonance imaging (MRI), nuclear magnetic resonance (NMR), computerized axial tomography (CAT) scan, cooled charged coupled device (CCD) camera optical imaging, bioluminescent optical imaging, position emission tomography (PET), single photon emission computed tomography, and microPET, thereby imaging a cancer cell in vivo.
42. The method of claim 41, wherein said mammal is a mouse, rat, . hamster, rabbit, pig, or human.
PCT/US2008/075291 2007-09-04 2008-09-04 High affinity anti-prostate stem cell antigen (psca) antibodies for cancer targeting and detection WO2009032949A2 (en)

Priority Applications (5)

Application Number Priority Date Filing Date Title
JP2010524150A JP6126773B2 (en) 2007-09-04 2008-09-04 High affinity anti-prostatic stem cell antigen (PSCA) antibody for cancer targeting and detection
US12/676,348 US8940298B2 (en) 2007-09-04 2008-09-04 High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection
EP08799192A EP2197491A4 (en) 2007-09-04 2008-09-04 High affinity anti-prostate stem cell antigen (psca) antibodies for cancer targeting and detection
CA2698343A CA2698343C (en) 2007-09-04 2008-09-04 High affinity anti-prostate stem cell antigen (psca) antibodies for cancer targeting and detection
US14/562,269 US9527919B2 (en) 2007-09-04 2014-12-05 High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US96993907P 2007-09-04 2007-09-04
US60/969,939 2007-09-04

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US12/676,348 A-371-Of-International US8940298B2 (en) 2007-09-04 2008-09-04 High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection
US14/562,269 Division US9527919B2 (en) 2007-09-04 2014-12-05 High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection

Publications (2)

Publication Number Publication Date
WO2009032949A2 true WO2009032949A2 (en) 2009-03-12
WO2009032949A3 WO2009032949A3 (en) 2009-12-30

Family

ID=40429695

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2008/075291 WO2009032949A2 (en) 2007-09-04 2008-09-04 High affinity anti-prostate stem cell antigen (psca) antibodies for cancer targeting and detection

Country Status (5)

Country Link
US (2) US8940298B2 (en)
EP (1) EP2197491A4 (en)
JP (3) JP6126773B2 (en)
CA (1) CA2698343C (en)
WO (1) WO2009032949A2 (en)

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2506876A2 (en) * 2009-12-02 2012-10-10 Imaginab, Inc. J591 minibodies and cys-diabodies for targeting human prostate specific membrane antigen (psma) and methods for their use
DE102011118022A1 (en) 2011-06-30 2013-01-03 Technische Universität Dresden Antibodies against the prostate-specific stem cell antigen and its use
CN104203975A (en) * 2011-06-23 2014-12-10 埃博灵克斯股份有限公司 Techniques for predicting, detecting and reducing non-specific protein interference in assays involving immunoglobulin single variable domains
US8940871B2 (en) 2006-03-20 2015-01-27 The Regents Of The University Of California Engineered anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting
US8940298B2 (en) 2007-09-04 2015-01-27 The Regents Of The University Of California High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection
WO2017062628A1 (en) * 2015-10-06 2017-04-13 City Of Hope Chimeric antigen receptors targeted to psca
US9701754B1 (en) 2002-10-23 2017-07-11 City Of Hope Covalent disulfide-linked diabodies and uses thereof
CN107602704A (en) * 2017-09-11 2018-01-19 贵州医科大学 A kind of preparation method of anti-prostate cancer stem cell antigen scfv fusion protein
US10035009B2 (en) 2013-04-15 2018-07-31 The Board Of Trustees Of The Leland Stanford Junior University Systems and methods for treating pancreatic cancer
US10287321B2 (en) 2011-03-17 2019-05-14 The University Of Birmingham Re-directed immunotherapy
US10517969B2 (en) 2009-02-17 2019-12-31 Cornell University Methods and kits for diagnosis of cancer and prediction of therapeutic value
US11254744B2 (en) 2015-08-07 2022-02-22 Imaginab, Inc. Antigen binding constructs to target molecules
US11266745B2 (en) 2017-02-08 2022-03-08 Imaginab, Inc. Extension sequences for diabodies
US11644471B2 (en) 2010-09-30 2023-05-09 Ablynx N.V. Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains
US11702482B2 (en) 2018-12-17 2023-07-18 Revitope Limited Twin immune cell engager
EP4028414A4 (en) * 2019-09-11 2024-01-17 Univ Pennsylvania Compositions and methods comprising prostate stem cell antigen (psca) chimeric antigen receptors (cars)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6136311A (en) 1996-05-06 2000-10-24 Cornell Research Foundation, Inc. Treatment and diagnosis of cancer
US6541212B2 (en) 1997-03-10 2003-04-01 The Regents Of The University Of California Methods for detecting prostate stem cell antigen protein
WO2012042026A1 (en) 2010-09-30 2012-04-05 Ablynx Nv Biological materials related to c-met
US20150344568A1 (en) * 2011-06-23 2015-12-03 Ablynx N.V. Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains
ES2759936T3 (en) 2011-06-23 2020-05-12 Ablynx Nv Serum albumin binding proteins
AU2019397048A1 (en) * 2018-12-13 2021-06-24 Bellicum Pharmaceuticals, Inc. PSCA CAR-T cells

Family Cites Families (252)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US121022A (en) 1871-11-14 Improvement in harness-fviountings
US943525A (en) 1908-01-04 1909-12-14 William Henry Drake Folding crate.
US5292668A (en) 1981-12-21 1994-03-08 Boston Biomedical Research Institute, Inc. Bispecific antibody determinants
US4444878A (en) 1981-12-21 1984-04-24 Boston Biomedical Research Institute, Inc. Bispecific antibody determinants
US5869620A (en) 1986-09-02 1999-02-09 Enzon, Inc. Multivalent antigen-binding proteins
US5260203A (en) 1986-09-02 1993-11-09 Enzon, Inc. Single polypeptide chain binding molecules
FR2604092B1 (en) 1986-09-19 1990-04-13 Immunotech Sa IMMUNOREACTIVES FOR TARGETING ANIMAL CELLS FOR VISUALIZATION OR DESTRUCTION IN VIVO
US5766960A (en) 1987-07-27 1998-06-16 Australian Membrane And Biotechnology Research Institute Receptor membranes
WO1989001159A1 (en) 1987-07-27 1989-02-09 Commonwealth Scientific And Industrial Research Or Receptor membranes
US4943525A (en) 1987-11-02 1990-07-24 Bioventures, Inc. Simultaneous immunoassay for the determination of antigens and antibodies
US4892824A (en) 1988-03-15 1990-01-09 Synbiotics Corporation Fast track method for producing monoclonal bi-specific immunoglobulins
US6010902A (en) 1988-04-04 2000-01-04 Bristol-Meyers Squibb Company Antibody heteroconjugates and bispecific antibodies for use in regulation of lymphocyte activity
US20030068322A1 (en) 1988-04-18 2003-04-10 Immunomedics, Inc. Methods of antibody-directed enzyme-prodrug therapy
US5851527A (en) 1988-04-18 1998-12-22 Immunomedics, Inc. Method for antibody targeting of therapeutic agents
ES2052027T5 (en) 1988-11-11 2005-04-16 Medical Research Council IMMUNOGLOBULINE VARIABLE DOMAIN SEQUENCE CLONING.
DE3920358A1 (en) 1989-06-22 1991-01-17 Behringwerke Ag BISPECIFIC AND OLIGO-SPECIFIC, MONO- AND OLIGOVALENT ANTI-BODY CONSTRUCTS, THEIR PRODUCTION AND USE
US6020145A (en) 1989-06-30 2000-02-01 Bristol-Myers Squibb Company Methods for determining the presence of carcinoma using the antigen binding region of monoclonal antibody BR96
IL94872A (en) 1989-06-30 1995-03-30 Oncogen Monoclonal or chimeric antibodies reactive with human carcinomas, recombinant proteins comprising their antigen binding region, pharmaceutical compositions and kits comprising said antibodies and methods for imaging human carcinoma using same
US5980896A (en) 1989-06-30 1999-11-09 Bristol-Myers Squibb Company Antibodies reactive with human carcinomas
US5332567A (en) 1989-08-24 1994-07-26 Immunomedics Detection and treatment of infections with immunoconjugates
US5582996A (en) 1990-12-04 1996-12-10 The Wistar Institute Of Anatomy & Biology Bifunctional antibodies and method of preparing same
US6106835A (en) 1991-04-19 2000-08-22 Tanox, Inc. Modified binding molecules specific for T or B lymphocytes and their use as in vivo immune modulators
US6197298B1 (en) 1991-04-19 2001-03-06 Tanox, Inc. Modified binding molecules specific for T lymphocytes and their use as in vivo immune modulators in animals
CA2065658A1 (en) 1991-04-19 1992-10-20 Tse-Wen Chang Conjugates of liposomes or microbeads and antibodies specific for t lymphocytes and their use in vivo immune modulators
US6129916A (en) 1991-04-19 2000-10-10 Tanox, Inc. Method of Increasing activation on proliferation of T cells using antibody-microbead conjugates
US5872222A (en) 1991-04-19 1999-02-16 Tanox Biosystems, Inc. Conjugates of polymers and antibodies specific for T lymphocytes, and their use as adjuvants
DE4118120A1 (en) 1991-06-03 1992-12-10 Behringwerke Ag TETRAVALENT BISPECIFIC RECEPTORS, THEIR PRODUCTION AND USE
US5770197A (en) 1991-06-27 1998-06-23 Bristol-Myers Squibb Company Methods for regulating the immune response using B7 binding molecules and IL4-binding molecules
US5851795A (en) 1991-06-27 1998-12-22 Bristol-Myers Squibb Company Soluble CTLA4 molecules and uses thereof
US6887471B1 (en) 1991-06-27 2005-05-03 Bristol-Myers Squibb Company Method to inhibit T cell interactions with soluble B7
EP0606217B2 (en) 1991-06-27 2008-12-03 Bristol-Myers Squibb Company Ctl4a receptor, fusion proteins containing it and uses thereof
US5844095A (en) 1991-06-27 1998-12-01 Bristol-Myers Squibb Company CTLA4 Ig fusion proteins
US6090914A (en) 1991-06-27 2000-07-18 Bristol-Myers Squibb Company CTLA4/CD28Ig hybrid fusion proteins and uses thereof
US5637481A (en) 1993-02-01 1997-06-10 Bristol-Myers Squibb Company Expression vectors encoding bispecific fusion proteins and methods of producing biologically active bispecific fusion proteins in a mammalian cell
DE4134982A1 (en) 1991-10-23 1993-04-29 Kernforschungsz Karlsruhe USE OF ANTIBODY-CONTAINING PREPARATIONS FOR IMMUNE SUPPRESSION
US5932448A (en) 1991-11-29 1999-08-03 Protein Design Labs., Inc. Bispecific antibody heterodimers
DE4140142A1 (en) 1991-12-05 1993-06-09 Boehringer Mannheim Gmbh, 6800 Mannheim, De MULTIVALENT DEXTRANREAGENT FOR USE IN PRECIPITATION TESTS
WO1993012232A1 (en) 1991-12-10 1993-06-24 Dana Farber Cancer Institute Reactive neutralizing human anti-gp120 recombinant antibody, dna coding the same and use thereof
FR2686899B1 (en) 1992-01-31 1995-09-01 Rhone Poulenc Rorer Sa NOVEL BIOLOGICALLY ACTIVE POLYPEPTIDES, THEIR PREPARATION AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM.
WO1993016185A2 (en) 1992-02-06 1993-08-19 Creative Biomolecules, Inc. Biosynthetic binding protein for cancer marker
US6093399A (en) 1992-03-05 2000-07-25 Board Of Regents, The University Of Texas System Methods and compositions for the specific coagulation of vasculature
US6749853B1 (en) 1992-03-05 2004-06-15 Board Of Regents, The University Of Texas System Combined methods and compositions for coagulation and tumor treatment
US5965132A (en) 1992-03-05 1999-10-12 Board Of Regents, The University Of Texas System Methods and compositions for targeting the vasculature of solid tumors
US6004555A (en) 1992-03-05 1999-12-21 Board Of Regents, The University Of Texas System Methods for the specific coagulation of vasculature
US5776427A (en) 1992-03-05 1998-07-07 Board Of Regents, The University Of Texas System Methods for targeting the vasculature of solid tumors
DE69332948T2 (en) 1992-03-05 2003-11-27 Univ Texas Use of immunoconjugates for the diagnosis and / or therapy of vascularized tumors
US5877289A (en) 1992-03-05 1999-03-02 The Scripps Research Institute Tissue factor compositions and ligands for the specific coagulation of vasculature
US6096289A (en) 1992-05-06 2000-08-01 Immunomedics, Inc. Intraoperative, intravascular, and endoscopic tumor and lesion detection, biopsy and therapy
SG41929A1 (en) 1992-09-25 1997-08-15 Commw Scient Ind Res Org Target binding polypeptide
WO1994008038A1 (en) 1992-10-02 1994-04-14 Trustees Of Dartmouth College Bispecific reagents for redirected targeting of low density lipoprotein
US5837821A (en) 1992-11-04 1998-11-17 City Of Hope Antibody construct
JP4204642B2 (en) 1992-11-05 2009-01-07 スローン − ケタリング・インスティテュート・フォー・キャンサー・リサーチ Prostate-specific membrane antigen
AU5670194A (en) 1992-11-20 1994-06-22 Enzon, Inc. Linker for linked fusion polypeptides
US5376249A (en) 1992-11-25 1994-12-27 Perseptive Biosystems, Inc. Analysis utilizing isoelectric focusing
GB9225453D0 (en) 1992-12-04 1993-01-27 Medical Res Council Binding proteins
CA2150262C (en) 1992-12-04 2008-07-08 Kaspar-Philipp Holliger Multivalent and multispecific binding proteins, their manufacture and use
ATE187494T1 (en) 1992-12-11 1999-12-15 Dow Chemical Co MULTIVALENT SINGLE CHAIN ANTIBODIES
US5861156A (en) 1993-01-08 1999-01-19 Creative Biomolecules Methods of delivering agents to target cells
US5773253A (en) 1993-01-22 1998-06-30 Bristol-Myers Squibb Company MYPPPY variants of CTL A4 and uses thereof
US6482919B2 (en) 1993-02-01 2002-11-19 Bristol-Myers Squibb Company Expression vectors encoding bispecific fusion proteins and methods of producing biologically active bispecific fusion proteins in a mammalian cell
ATE241642T1 (en) 1993-02-04 2003-06-15 Denzyme Aps IMPROVED METHOD FOR REFOLDING PROTEINS
US5705614A (en) 1993-04-09 1998-01-06 Chiron Corporation Methods of producing antigen forks
CN100341896C (en) 1993-09-02 2007-10-10 达特茅斯学院理事 Anti-GP39 antibodies and uses therefor
US5869049A (en) 1993-09-02 1999-02-09 Trustees Of Dartmouth College Methods of inducing T cell unresponsiveness to bone marrow with gp39 antagonists
ZA946765B (en) 1993-09-02 1996-02-15 Dartmouth College Methods of prolonged suppression of humoral immunity
EP0720624B1 (en) 1993-09-22 1998-11-25 Medical Research Council Retargeting antibodies
GB9324807D0 (en) 1993-12-03 1994-01-19 Cancer Res Campaign Tech Tumour antibody
DE4421391C1 (en) 1994-06-18 1995-11-30 Gsf Forschungszentrum Umwelt Use of antibodies against T cells for prolonged immunosuppression
US5846782A (en) 1995-11-28 1998-12-08 Genvec, Inc. Targeting adenovirus with use of constrained peptide motifs
US5965541A (en) 1995-11-28 1999-10-12 Genvec, Inc. Vectors and methods for gene transfer to cells
US5962311A (en) 1994-09-08 1999-10-05 Genvec, Inc. Short-shafted adenoviral fiber and its use
US6465253B1 (en) 1994-09-08 2002-10-15 Genvec, Inc. Vectors and methods for gene transfer to cells
US5559099A (en) 1994-09-08 1996-09-24 Genvec, Inc. Penton base protein and methods of using same
US5541087A (en) 1994-09-14 1996-07-30 Fuji Immunopharmaceuticals Corporation Expression and export technology of proteins as immunofusins
US5688690A (en) 1994-09-16 1997-11-18 The Wistar Institute Of Anatomy And Biology Human cytotoxic lymphocyte signal transduction surface protein (P38) and monoclonal antibodies thereto
JP2001526522A (en) 1995-02-24 2001-12-18 スローン−ケタリング・インスティテュート・フォー・キャンサー・リサーチ Prostate specific membrane antigen and uses thereof
US5731168A (en) 1995-03-01 1998-03-24 Genentech, Inc. Method for making heteromultimeric polypeptides
GB9504344D0 (en) 1995-03-03 1995-04-19 Unilever Plc Antibody fragment production
US5837281A (en) * 1995-03-17 1998-11-17 Takeda Chemical Industries, Ltd. Stabilized interface for iontophoresis
US6096871A (en) 1995-04-14 2000-08-01 Genentech, Inc. Polypeptides altered to contain an epitope from the Fc region of an IgG molecule for increased half-life
US5747035A (en) 1995-04-14 1998-05-05 Genentech, Inc. Polypeptides with increased half-life for use in treating disorders involving the LFA-1 receptor
US6121022A (en) 1995-04-14 2000-09-19 Genentech, Inc. Altered polypeptides with increased half-life
AUPO591797A0 (en) 1997-03-27 1997-04-24 Commonwealth Scientific And Industrial Research Organisation High avidity polyvalent and polyspecific reagents
US5830478A (en) 1995-06-07 1998-11-03 Boston Biomedical Research Institute Method for delivering functional domains of diphtheria toxin to a cellular target
CN1173878A (en) 1995-10-16 1998-02-18 尤尼利弗公司 Bifunctional or bivalent antibody fragment analogue
ATE265532T1 (en) 1995-10-19 2004-05-15 Bristol Myers Squibb Co MONOCLONAL ANTIBODY BR110 AND ITS APPLICATION
DK0914155T3 (en) 1996-03-25 2006-06-26 Medarex Inc Monoclonal antibodies specific for the extracellular domain of prostate-specific membrane antigen
US6962981B1 (en) 1996-03-25 2005-11-08 Medarex, Inc. Monoclonal antibodies specific for the extracellular domain of prostate-specific membrane antigen
US6150508A (en) 1996-03-25 2000-11-21 Northwest Biotherapeutics, Inc. Monoclonal antibodies specific for the extracellular domain of prostate-specific membrane antigen
US7381407B1 (en) 1996-03-25 2008-06-03 Medarex, Inc. Monoclonal antibodies specific for the extracellular domain of prostate-specific membrane antigen
US6136311A (en) 1996-05-06 2000-10-24 Cornell Research Foundation, Inc. Treatment and diagnosis of cancer
AU2961397A (en) 1996-05-23 1997-12-09 Inverness Medical Switzerland Gmbh Improvements in or relating to specific binding assays
AU729029B2 (en) 1996-07-03 2001-01-25 Genentech Inc. Hepatocyte growth factor receptor agonists and uses thereof
GB9712818D0 (en) 1996-07-08 1997-08-20 Cambridge Antibody Tech Labelling and selection of specific binding molecules
US5922845A (en) 1996-07-11 1999-07-13 Medarex, Inc. Therapeutic multispecific compounds comprised of anti-Fcα receptor antibodies
US6653104B2 (en) 1996-10-17 2003-11-25 Immunomedics, Inc. Immunotoxins, comprising an internalizing antibody, directed against malignant and normal cells
CA2269060C (en) 1996-10-17 2009-04-07 Immunomedics, Inc. Non-antigenic toxin-conjugate and fusion protein of internalizing receptor system
EP1249705A3 (en) 1996-12-31 2003-11-05 Genometrix Genomics Incorporated Multiplexed molecular analysis apparatus and its fabrication method
US6258939B1 (en) 1997-03-10 2001-07-10 The Regents Of The University Of California PSCA antibodies and hybridomas producing them
US20080318254A9 (en) 1997-03-10 2008-12-25 The Regents Of The University Of California PSCA antibodies and hybridomas producing them
CA2281877C (en) 1997-03-10 2010-01-05 The Regents Of The University Of California Psca: prostate stem cell antigen
US6261789B1 (en) * 1997-03-10 2001-07-17 The Regents Of The University Of California Methods for detecting the presence of a PSCA protein using PSCA antibodies
US6541212B2 (en) 1997-03-10 2003-04-01 The Regents Of The University Of California Methods for detecting prostate stem cell antigen protein
US6261791B1 (en) 1997-03-10 2001-07-17 The Regents Of The University Of California Method for diagnosing cancer using specific PSCA antibodies
US6960443B2 (en) 1997-03-10 2005-11-01 The Regents Of The University Of California Methods of detecting prostate stem cell antigen in cancer
AU6914098A (en) 1997-04-03 1998-10-30 Universite Laval Transgenic expression in genital tract and sexual accessory glands
CA2288232A1 (en) 1997-05-02 1998-11-12 The Government Of The United States, Represented By The Secretary, Depar Tment Of Health And Human Services Immunotoxins directed against malignant cells
AU8296098A (en) 1997-07-08 1999-02-08 Board Of Regents, The University Of Texas System Compositions and methods for homoconjugates of antibodies which induce growth arrest or apoptosis of tumor cells
US6977074B2 (en) 1997-07-10 2005-12-20 Mannkind Corporation Method of inducing a CTL response
US6994851B1 (en) 1997-07-10 2006-02-07 Mannkind Corporation Method of inducing a CTL response
DE69837095T2 (en) 1997-09-03 2007-11-22 Immunomedics, Inc. FLUORATION OF PROTEINS AND PEPTIDES FOR POSITRONOMISSION STOMOGRAPHY
US6030792A (en) 1997-11-13 2000-02-29 Pfizer Inc Assays for measurement of protein fragments in biological media
US6709844B1 (en) 2000-11-16 2004-03-23 Mannkind Corporation Avoidance of undesirable replication intermediates in plasmid propagation
DE19911329A1 (en) 1998-03-27 2000-09-21 Benes Ivan Friedrich Radioimmunoconjugate which can be used in human therapy and process for its preparation
CZ121599A3 (en) 1998-04-09 1999-10-13 Aventis Pharma Deutschland Gmbh Single-chain molecule binding several antigens, process of its preparation and medicament in which the molecule is comprised
DK2180007T4 (en) 1998-04-20 2017-11-27 Roche Glycart Ag Glycosylation technique for antibodies to enhance antibody-dependent cell cytotoxicity
US6200765B1 (en) 1998-05-04 2001-03-13 Pacific Northwest Cancer Foundation Non-invasive methods to detect prostate cancer
US6071490A (en) 1998-05-07 2000-06-06 Immunomedics, Inc. Position emission tomography using gallium-68 chelates
WO1999059633A1 (en) 1998-05-20 1999-11-25 Immunomedics, Inc. Therapeutics using a bispecific anti-hla class ii invariant chain x anti-pathogen antibody
US20060052321A1 (en) 2002-04-05 2006-03-09 Raitano Arthur B Nucleic acid and corresponding protein entitled 98P4B6 useful in treatment and detection of cancer
US20040141975A1 (en) 1998-06-01 2004-07-22 Raitano Arthur B. Nucleic acid and corresponding protein entitled 98P4B6 useful in treatment and detection of cancer
DK1086223T3 (en) 1998-06-01 2009-11-30 Agensys Inc Novel serpentine transmembrane antigens expressed in human cancers and their applications
CA2331789C (en) 1998-07-13 2013-09-10 Board Of Regents, The University Of Texas System Cancer treatment methods using therapeutic conjugates that bind to aminophospholipids
WO2000014234A1 (en) 1998-09-08 2000-03-16 Urocor, Inc. Prostate specific promoter and regulation of gene expression
US6361774B1 (en) 1999-09-17 2002-03-26 Immunomedics, Inc. Methods and compositions for increasing the target-specific toxicity of a chemotherapy drug
JP2002538767A (en) 1998-09-29 2002-11-19 ザ ユーエイビー リサーチ ファンデイション Immunomodulation by genetic modification of dendritic cells and B cells
US6642007B1 (en) 1998-11-02 2003-11-04 Pfizer Inc. Assays for measurement of type II collagen fragments in urine
WO2000034461A2 (en) 1998-12-10 2000-06-15 Board Of Regents, The University Of Texas System Compositions and methods of modulating cholesterol metabolism
EP1918303A3 (en) * 1999-04-09 2008-05-14 Universität Zürich method for the stabilization of chimeric immunoglobulins or immunoglobulin fragments, and stabilized anti-egp-2 scfv fragment
EP1175616A4 (en) 1999-04-13 2003-08-13 Northwest Biotherapeutics Inc Methods for the diagnosis and treatment of metastatic prostate tumors
ES2223705T3 (en) 1999-04-28 2005-03-01 Board Of Regents, The University Of Texas System COMPOSITIONS AND METHODS FOR THE TREATMENT OF CANCER THROUGH INHIBITION SELECTIVE OF VEGF.
US6703020B1 (en) 1999-04-28 2004-03-09 Board Of Regents, The University Of Texas System Antibody conjugate methods for selectively inhibiting VEGF
GB9915653D0 (en) 1999-07-06 1999-09-01 Bruce Terry Hose clamp
WO2001009303A2 (en) 1999-07-30 2001-02-08 Vical Inc. Flt-3 LIGAND-ENCODING POLYNUCLEOTIDE AS A POLYNUCLEOTIDE-BASED VACCINE ENHANCER
KR20050004240A (en) 1999-08-31 2005-01-12 제넨테크, 인크. Compositions and Methods for the Treatment of Tumor
US6346249B1 (en) 1999-10-22 2002-02-12 Ludwig Institute For Cancer Research Methods for reducing the effects of cancers that express A33 antigen using A33 antigen specific immunoglobulin products
US6342587B1 (en) 1999-10-22 2002-01-29 Ludwig Institute For Cancer Research A33 antigen specific immunoglobulin products and uses thereof
AU778199B2 (en) 1999-10-29 2004-11-25 Genentech Inc. Anti-prostate stem cell antigen (PSCA) antibody compositions and methods of use
US6824780B1 (en) * 1999-10-29 2004-11-30 Genentech, Inc. Anti-tumor antibody compositions and methods of use
EP1266009B1 (en) * 2000-02-25 2008-12-31 THE GOVERNMENT OF THE UNITED STATES OF AMERICA, as represented by THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES ANTI-EGFRvIII SCFVS WITH IMPROVED CYTOTOXICITY AND YIELD, IMMUNOTOXINS BASED THEREON, AND METHODS OF USE THEREOF
WO2001071005A2 (en) 2000-03-24 2001-09-27 Micromet Ag Multifunctional polypeptides comprising a binding site to an epitope of the nkg2d receptor complex
US6573096B1 (en) 2000-04-01 2003-06-03 The Research Foundation At State University Of New York Compositions and methods for inhibition of cancer invasion and angiogenesis
IL151853A0 (en) 2000-04-11 2003-04-10 Genentech Inc Multivalent antibodies and uses therefor
US6861234B1 (en) 2000-04-28 2005-03-01 Mannkind Corporation Method of epitope discovery
EP1276896B1 (en) 2000-04-28 2010-07-21 Mannkind Corporation Epitope synchronization in antigen presenting cells
US6375991B1 (en) 2000-09-08 2002-04-23 Albemarle Corporation Production of concentrated biocidal solutions
DE10045591A1 (en) 2000-09-15 2002-04-04 Klaus Pfizenmaier Site-specific, antibody-mediated activation of proapoptotic cytokines - AMAIZe (Antibody-Mediated Apoptosis Inducing Cytokines)
WO2003008537A2 (en) 2001-04-06 2003-01-30 Mannkind Corporation Epitope sequences
KR20040054669A (en) 2001-08-03 2004-06-25 글리카트 바이오테크놀로지 아게 Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity
US7494646B2 (en) 2001-09-06 2009-02-24 Agensys, Inc. Antibodies and molecules derived therefrom that bind to STEAP-1 proteins
JP2005505271A (en) 2001-09-06 2005-02-24 アジェンシス， インコーポレイテッド Nucleic acids and corresponding proteins given the name STEAP-1 useful in the treatment and detection of cancer
CN100423777C (en) 2001-10-25 2008-10-08 杰南技术公司 Glycoprotein compositions
ES2358642T3 (en) 2001-11-07 2011-05-12 Mannkind Corporation EXPRESSION VECTORS CODING ANTIGENS EPITHOPES AND METHODS FOR THEIR DESIGN.
US20040018519A1 (en) 2001-11-16 2004-01-29 Wright ,Jr. George L Methods and devices for quantitative detection of prostate specific membrane antigen and other prostatic markers
WO2003050140A1 (en) 2001-12-10 2003-06-19 Kyogo Itoh Tumor antigens
AU2002351353A1 (en) 2001-12-19 2003-09-02 Genentech, Inc. Stabilizing polypeptides which have been exposed to urea
EP2048154B2 (en) 2002-02-05 2015-06-03 Genentech, Inc. Protein purification
JP4319979B2 (en) 2002-04-26 2009-08-26 ジェネンテック・インコーポレーテッド Non-affinity purification of proteins
JP2005530856A (en) 2002-06-21 2005-10-13 ジョンズ ホプキンス ユニバーシティー スクール オブ メディシン Membrane-related tumor endothelial marker
US9809654B2 (en) 2002-09-27 2017-11-07 Vaccinex, Inc. Targeted CD1d molecules
US7820166B2 (en) 2002-10-11 2010-10-26 Micromet Ag Potent T cell modulating molecules
US9701754B1 (en) 2002-10-23 2017-07-11 City Of Hope Covalent disulfide-linked diabodies and uses thereof
US7449616B2 (en) * 2002-12-24 2008-11-11 Pfizer Inc. Anti-NGF antibodies and methods using same
EP2264151B1 (en) 2003-01-22 2016-04-20 Roche Glycart AG Fusion constructs and use of same to produce antibodies with increased FC receptor binding affinity and effector function
US7453436B2 (en) * 2003-03-21 2008-11-18 Ruiz David M Hand-held on-screen control device
WO2004088279A2 (en) 2003-03-28 2004-10-14 Iowa State University Research Foundation, Inc. Allosteric probes and methods
GB0308988D0 (en) 2003-04-17 2003-05-28 Univ Singapore Molecule
US7595379B2 (en) 2003-05-30 2009-09-29 Agensys, Inc. Antibodies and related molecules that bind to PSCA proteins
US7541442B2 (en) 2003-05-30 2009-06-02 Agensys, Inc. Antibodies and related molecules that bind to PSCA proteins
US7622564B2 (en) 2003-05-30 2009-11-24 Agensys, Inc. Prostate stem cell antigen (PSCA) variants and subsequences thereof
US8784821B1 (en) 2003-05-31 2014-07-22 Amgen Research (Munich) Gmbh Human-anti-human cd3 binding molecules
AU2004252171B2 (en) 2003-06-27 2011-04-21 Biogen Ma Inc. Modified binding molecules comprising connecting peptides
SI1648940T1 (en) 2003-07-28 2016-08-31 Genentech, Inc. Reducing protein a leaching during protein a affinity chromatography
GB0321805D0 (en) 2003-09-18 2003-10-15 Univ Wales Medicine Human tumour growth patterns
US7524813B2 (en) 2003-10-10 2009-04-28 Novo Nordisk Health Care Ag Selectively conjugated peptides and methods of making the same
BRPI0415457A (en) 2003-10-16 2006-12-05 Micromet Ag cytotoxically active cd3 specific binding construct, its production process, composition comprising the same, nucleic acid sequence, vector, host, its uses in the preparation of a pharmaceutical composition and kit comprising the same
ES2428358T3 (en) 2003-10-17 2013-11-07 Novo Nordisk A/S Combination therapy
ITPD20030264A1 (en) 2003-10-30 2005-04-30 Xeptagen Spa HIGHLY SPECIFIC DIAGNOSIS METHOD FOR NEOPLASIA
KR101192496B1 (en) 2003-11-06 2012-10-18 시애틀 지네틱스, 인크. Monomethylvaline compounds capable of conjugation to ligands
US7235641B2 (en) 2003-12-22 2007-06-26 Micromet Ag Bispecific antibodies
ATE553189T1 (en) 2004-01-16 2012-04-15 Fraunhofer Ges Forschung IMMUNOKINASES
CA2555503C (en) 2004-02-16 2018-03-27 Micromet Ag Humanized anti-cd3 bispecific binding molecules having reduced immunogenicity, uses and compositions relating thereto
JP5064037B2 (en) 2004-02-23 2012-10-31 ジェネンテック， インコーポレイテッド Heterocyclic self-destructive linkers and conjugates
US7662936B2 (en) 2004-04-07 2010-02-16 Genentech, Inc. Mass spectrometry of antibody conjugates
CA2568238C (en) 2004-04-12 2013-10-15 Medical College Of Ohio Methods and compositions for assaying analytes
BRPI0418766B8 (en) 2004-04-22 2021-05-25 Agensys Inc antibody or fragment thereof, vector, pharmaceutical composition, assay and method for detecting the presence of step-1 protein, as well as a method for delivering a cytotoxic agent or a diagnostic agent
CN114053429A (en) 2004-06-01 2022-02-18 健泰科生物技术公司 Antibody-drug conjugates and methods
US20080227736A1 (en) 2004-06-03 2008-09-18 Regents Of The University Of California, Targeting Pseudotyped Retroviral Vectors
US20060008468A1 (en) 2004-06-17 2006-01-12 Chih-Sheng Chiang Combinations of tumor-associated antigens in diagnostics for various types of cancers
US20060159689A1 (en) 2004-06-17 2006-07-20 Chih-Sheng Chiang Combinations of tumor-associated antigens in diagnostics for various types of cancers
US7323682B2 (en) * 2004-07-02 2008-01-29 Thermo Finnigan Llc Pulsed ion source for quadrupole mass spectrometer and method
MX2007000387A (en) 2004-07-16 2007-03-28 Micromet Ag Expression-enhanced polypeptides.
RU2412947C2 (en) 2004-09-23 2011-02-27 Дженентек, Инк. Antibodies, constructed on cysteine basis and their conjugates
US20100111856A1 (en) 2004-09-23 2010-05-06 Herman Gill Zirconium-radiolabeled, cysteine engineered antibody conjugates
GB0422431D0 (en) * 2004-10-08 2004-11-10 Affitech As Method
US7947839B2 (en) 2004-12-01 2011-05-24 Genentech, Inc. Heterocyclic-substituted bis-1,8 naphthalimide compounds, antibody drug conjugates, and methods of use
US20070134243A1 (en) 2004-12-01 2007-06-14 Gazzard Lewis J Antibody drug conjugates and methods
CA2491067A1 (en) 2004-12-24 2006-06-24 Stichting Katholieke Universiteit Mrna rations in urinary sediments and/or urine as a prognostic marker for prostate cancer
EP1874822A1 (en) 2005-04-14 2008-01-09 Agensys, Inc. Antibodies and related molecules that bind to psca proteins
US20060246524A1 (en) 2005-04-28 2006-11-02 Christina Bauer Nanoparticle conjugates
US8088908B2 (en) 2005-05-10 2012-01-03 City Of Hope Humanized anti-prostate stem cell antigen monoclonal antibody
GB2426581A (en) 2005-05-27 2006-11-29 Univ Nottingham Immunoassay methods
ES2539250T3 (en) 2005-07-25 2015-06-29 Emergent Product Development Seattle, Llc Reduction of B cells through the use of CD37 specific binding and CD20 specific binding molecules
US7612181B2 (en) 2005-08-19 2009-11-03 Abbott Laboratories Dual variable domain immunoglobulin and uses thereof
US20090226465A1 (en) 2005-10-31 2009-09-10 Jackson David Y Macrocyclic depsipeptide antibody-drug conjugates and methods
US20070212331A1 (en) 2006-03-07 2007-09-13 Baldassare Joseph J Methods and compositions for selectively killing cells
ES2363891T3 (en) * 2006-03-20 2011-08-18 The Regents Of The University Of California ANTIBODIES AGAINST THE ANTIGEN OF TRONCAL CELLS OF THE PROSTATE (PSCA) GENETICALLY MODIFIED FOR ADDRESSING TO CANCER.
BRPI0708970A2 (en) 2006-03-21 2011-06-21 Wyeth Corp method for treating a subject with a disease or disorder characterized by amyloid deposition of a-beta; method of inhibiting or reducing the accumulation of amyloid deposits of a-beta in a subject; method of inhibiting or reducing neurodegeneration in a subject; and method of inhibiting or reducing cognitive decline, or improving cognition, in a subject
JP2009534309A (en) 2006-03-31 2009-09-24 マサチューセッツ インスティテュート オブ テクノロジー System for targeted delivery of therapeutic agents
US7566276B2 (en) 2006-04-14 2009-07-28 Dogleg Right Corporation Multi-piece putter head having an insert
US20110052697A1 (en) 2006-05-17 2011-03-03 Gwangju Institute Of Science & Technology Aptamer-Directed Drug Delivery
US8574848B2 (en) 2006-09-13 2013-11-05 Oncimmune Ltd. Immunoassay methods
RU2450832C2 (en) 2006-10-25 2012-05-20 Конинклейке Филипс Электроникс Н.В. Radiopaque substances for prostate cancer detection
US8163886B2 (en) 2006-12-21 2012-04-24 Emd Millipore Corporation Purification of proteins
US8362217B2 (en) 2006-12-21 2013-01-29 Emd Millipore Corporation Purification of proteins
EP2144930A1 (en) 2007-04-18 2010-01-20 ZymoGenetics, Inc. Single chain fc, methods of making and methods of treatment
WO2009003493A2 (en) 2007-07-03 2009-01-08 Dako Denmark A/S Mhc multimers, methods for their generation, labeling and use
CA2698343C (en) * 2007-09-04 2018-06-12 The Regents Of The University Of California High affinity anti-prostate stem cell antigen (psca) antibodies for cancer targeting and detection
WO2009039854A2 (en) 2007-09-27 2009-04-02 Dako Denmark A/S Mhc multimers in tuberculosis diagnostics, vaccine and therapeutics
CA2702586C (en) 2007-10-18 2017-08-01 Bn Immunotherapeutics Inc. Use of mva to treat prostate cancer
US20090169613A1 (en) 2007-11-09 2009-07-02 Reznik Boris N Targeting of tumor stem cells through selective silencing of boris expression
US9107858B2 (en) 2007-12-05 2015-08-18 Wisconsin Alumni Research Foundation Dendritic cell targeting compositions and uses thereof
US8043830B2 (en) 2008-02-01 2011-10-25 The Regents Of The University Of California Biotin-ligase system for secretion of biotinylated protein
EP2240495B1 (en) 2008-02-01 2015-07-15 Genentech, Inc. Nemorubicin metabolite and analog reagents, antibody-drug conjugates and methods
TW200947474A (en) 2008-05-05 2009-11-16 Ren-Huan Pan Manufacturing equipment and method of quadratic arc alignment dua-turret formation center, and miniature milliohm current sensors manufactured thereof
AU2009246516B2 (en) 2008-05-13 2015-03-05 Genentech, Inc. Analysis of antibody drug conjugates by bead-based affinity capture and mass spectrometry
EP2303332B1 (en) 2008-07-15 2014-12-31 Genentech, Inc. Anthracycline conjugates, process for their preparation and their use as antitumor compounds
US20100069616A1 (en) 2008-08-06 2010-03-18 The Regents Of The University Of California Engineered antibody-nanoparticle conjugates
WO2010027629A2 (en) 2008-09-08 2010-03-11 Conocophillips Company System for incondensable component separation in a liquefied natural gas facility
WO2010037408A1 (en) 2008-09-30 2010-04-08 Curevac Gmbh Composition comprising a complexed (m)rna and a naked mrna for providing or enhancing an immunostimulatory response in a mammal and uses thereof
WO2010037835A2 (en) 2008-10-01 2010-04-08 Micromet Ag Cross-species-specific pscaxcd3, cd19xcd3, c-metxcd3, endosialinxcd3, epcamxc d3, igf-1rxcd3 or fapalpha xcd3 bispecific single chain antibody
WO2010037397A1 (en) 2008-10-01 2010-04-08 Dako Denmark A/S Mhc multimers in cmv immune monitoring
US20110318380A1 (en) 2008-10-01 2011-12-29 Dako Denmark A/S MHC Multimers in Cancer Vaccines and Immune Monitoring
US20110207155A1 (en) 2008-10-13 2011-08-25 Xeptagen Spa Method for the preparation of immunoconjugates and use thereof
JP2012507300A (en) 2008-10-30 2012-03-29 カリス ライフ サイエンシズ ルクセンブルク ホールディングス Method for evaluating RNA patterns
US7897356B2 (en) 2008-11-12 2011-03-01 Caris Life Sciences Methods and systems of using exosomes for determining phenotypes
CA2743487A1 (en) 2008-11-13 2010-05-20 Emergent Product Development Seattle, Llc Cd37 immunotherapeutic combination therapies and uses thereof
WO2010074702A1 (en) 2008-12-16 2010-07-01 Millipore Corporation Purification of proteins
CN102265059B (en) 2008-12-25 2013-08-07 东海橡塑工业株式会社 Fluid-sealed vibration damping device
WO2010081173A2 (en) 2009-01-12 2010-07-15 Cytomx Therapeutics, Llc Modified antibody compositions, methods of making and using thereof
WO2010102195A2 (en) 2009-03-06 2010-09-10 The Johns Hopkins University Annexin a11 and associated genes as biomarkers for cancer
WO2010111388A2 (en) 2009-03-24 2010-09-30 Biocept, Inc. Devices and methods of cell capture and analysis
WO2010129304A2 (en) 2009-04-27 2010-11-11 Oncomed Pharmaceuticals, Inc. Method for making heteromultimeric molecules
US20100303821A1 (en) 2009-05-28 2010-12-02 Glaxo Group Limited Immunoglobulins
AU2010268690B2 (en) 2009-07-03 2011-11-10 Avipep Pty Ltd Immuno-conjugates and methods for producing them
TWM383852U (en) 2009-07-13 2010-07-01 Speedtech Corp An improvement of the universal serial bus connector
CN102573925B (en) 2009-10-05 2015-08-05 佳能株式会社 The acousto-optic imaging method of photoacoustic imaging contrast agent and the described photoacoustic imaging contrast agent of use
CA2782333C (en) 2009-12-02 2019-06-04 Imaginab, Inc. J591 minibodies and cys-diabodies for targeting human prostate specific membrane antigen (psma) and methods for their use
US20110142811A1 (en) 2009-12-16 2011-06-16 Deutsches Krebsforschungszentrum Measles virus for the elimination of unwanted cell populations
US9315581B2 (en) 2009-12-23 2016-04-19 A Vipep Pty Limited Immuno-conjugates and methods for producing them
PT2519543T (en) 2009-12-29 2016-10-07 Emergent Product Dev Seattle Heterodimer binding proteins and uses thereof
EP2542696B1 (en) 2010-03-01 2016-09-28 Caris Life Sciences Switzerland Holdings GmbH Biomarkers for theranostics

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of EP2197491A4 *

Cited By (36)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9765155B2 (en) 2002-10-23 2017-09-19 City Of Hope Covalent disulfide-linked diabodies and uses thereof
US9701754B1 (en) 2002-10-23 2017-07-11 City Of Hope Covalent disulfide-linked diabodies and uses thereof
US8940871B2 (en) 2006-03-20 2015-01-27 The Regents Of The University Of California Engineered anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting
US9527919B2 (en) 2007-09-04 2016-12-27 The Regents Of The University Of California High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection
US8940298B2 (en) 2007-09-04 2015-01-27 The Regents Of The University Of California High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection
US10517969B2 (en) 2009-02-17 2019-12-31 Cornell University Methods and kits for diagnosis of cancer and prediction of therapeutic value
EP2506876A4 (en) * 2009-12-02 2013-07-31 Imaginab Inc J591 minibodies and cys-diabodies for targeting human prostate specific membrane antigen (psma) and methods for their use
US8772459B2 (en) 2009-12-02 2014-07-08 Imaginab, Inc. J591 minibodies and Cys-diabodies for targeting human prostate specific membrane antigen (PSMA) and methods for their use
EP2506876A2 (en) * 2009-12-02 2012-10-10 Imaginab, Inc. J591 minibodies and cys-diabodies for targeting human prostate specific membrane antigen (psma) and methods for their use
AU2010325969B2 (en) * 2009-12-02 2016-10-20 Imaginab, Inc. J591 minibodies and cys-diabodies for targeting human prostate specific membrane antigen
US11180570B2 (en) 2009-12-02 2021-11-23 Imaginab, Inc. J591 minibodies and cys-diabodies for targeting human prostate specific membrane antigen (PSMA) and methods for their use
CN102753194A (en) * 2009-12-02 2012-10-24 伊麦吉纳博公司 J591 minibodies and cys-diabodies for targeting human prostate specific membrane antigen (psma) and methods for their use
US11644471B2 (en) 2010-09-30 2023-05-09 Ablynx N.V. Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains
US10287321B2 (en) 2011-03-17 2019-05-14 The University Of Birmingham Re-directed immunotherapy
US11236131B2 (en) 2011-03-17 2022-02-01 The University Of Birmingham Re-directed immunotherapy
CN104203975A (en) * 2011-06-23 2014-12-10 埃博灵克斯股份有限公司 Techniques for predicting, detecting and reducing non-specific protein interference in assays involving immunoglobulin single variable domains
US11192937B2 (en) 2011-06-23 2021-12-07 Ablynx N.V. Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains
US11192938B2 (en) 2011-06-23 2021-12-07 Ablynx N.V. Serum albumin binding proteins containing immunoglobulin single variable domains
US10858418B2 (en) 2011-06-23 2020-12-08 Ablynx N.V. Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains
US9200078B2 (en) 2011-06-30 2015-12-01 Gemoab Monoclonals Gmbh Antibodies against prostate-specific stem cell antigen and use thereof
DE102011118022B4 (en) * 2011-06-30 2018-01-18 Gemoab Monoclonals Gmbh Antibodies against the prostate-specific stem cell antigen and its use
DE102011118022A1 (en) 2011-06-30 2013-01-03 Technische Universität Dresden Antibodies against the prostate-specific stem cell antigen and its use
EP3505536A1 (en) * 2011-06-30 2019-07-03 GEMoaB Monoclonals GmbH Antibodies directed against the prostate-specific stem cell antigen and its use
WO2013001065A1 (en) 2011-06-30 2013-01-03 Technische Universität Dresden Antibodies against prostate-specific stem cell antigen and use thereof
US10035009B2 (en) 2013-04-15 2018-07-31 The Board Of Trustees Of The Leland Stanford Junior University Systems and methods for treating pancreatic cancer
US11254744B2 (en) 2015-08-07 2022-02-22 Imaginab, Inc. Antigen binding constructs to target molecules
WO2017062628A1 (en) * 2015-10-06 2017-04-13 City Of Hope Chimeric antigen receptors targeted to psca
EP3747462A1 (en) * 2015-10-06 2020-12-09 City of Hope Chimeric antigen receptors targeted to psca
US11466097B2 (en) 2015-10-06 2022-10-11 City Of Hope Chimeric antigen receptors targeted to PSCA
IL258502B2 (en) * 2015-10-06 2023-05-01 Hope City Chimeric antigen receptors targeted to psca
IL258502A (en) * 2015-10-06 2018-05-31 Hope City Chimeric antigen receptors targeted to psca
AU2016336515B2 (en) * 2015-10-06 2023-08-24 City Of Hope Chimeric antigen receptors targeted to PSCA
US11266745B2 (en) 2017-02-08 2022-03-08 Imaginab, Inc. Extension sequences for diabodies
CN107602704A (en) * 2017-09-11 2018-01-19 贵州医科大学 A kind of preparation method of anti-prostate cancer stem cell antigen scfv fusion protein
US11702482B2 (en) 2018-12-17 2023-07-18 Revitope Limited Twin immune cell engager
EP4028414A4 (en) * 2019-09-11 2024-01-17 Univ Pennsylvania Compositions and methods comprising prostate stem cell antigen (psca) chimeric antigen receptors (cars)

Also Published As

Publication number Publication date
EP2197491A2 (en) 2010-06-23
CA2698343A1 (en) 2009-03-12
JP2010538080A (en) 2010-12-09
JP6126773B2 (en) 2017-05-10
JP2017158554A (en) 2017-09-14
US20150239983A1 (en) 2015-08-27
WO2009032949A3 (en) 2009-12-30
JP2015015960A (en) 2015-01-29
US8940298B2 (en) 2015-01-27
CA2698343C (en) 2018-06-12
US9527919B2 (en) 2016-12-27
EP2197491A4 (en) 2011-01-12
US20100297004A1 (en) 2010-11-25
JP6120819B2 (en) 2017-04-26

Similar Documents

Publication Publication Date Title
US9527919B2 (en) High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection
JP6856611B2 (en) Antigen-binding construct for CD8
CN107921121B (en) Antibody therapeutics that bind CD47
US20200255537A1 (en) Internalizing human monoclonal antibodies targeting prostate cancer cells in situ
Olafsen et al. Optimizing radiolabeled engineered anti-p185HER2 antibody fragments for in vivo imaging
KR100620554B1 (en) -72 Humanized Anti-TAG-72 Monoclonal Antibodies
AU2008234615B2 (en) The ED-A antigen of fibrinogen is associated with the neovasculature of tumour metastases
JP5693779B2 (en) Antibodies against endoplasmin and uses thereof
US20150191543A1 (en) Engineered antibody fragments for targeting and imaging cd8 expression in vivo
US20080287309A1 (en) Methods for Discovering Antibodies Specific to Cancer Cells and Antibodies Discovered Thereby
CN115960230A (en) Antigen binding constructs for targeting molecules
JP6990652B2 (en) New polypeptide
CN104768563B (en) Antigens associated with inflammatory bowel disease
WO2018166507A1 (en) Novel recombinant bifunctional fusion protein, preparation method therefor and use thereof
JP2013538042A5 (en)
JP5881085B2 (en) Composition for treatment and diagnosis of pancreatic cancer
JP2024505001A (en) Compositions and methods for treating and preventing diseases associated with AVB8 integrin
CA2490659C (en) Humanized anti-tag-72 cc49 for diagnosis and therapy of human tumors
Albrecht et al. Update: Recombinant antibodies: From the laboratory to the clinic
US20230181772A1 (en) Engineered anti-prostate stem cell antigen fusion proteins and uses thereof
WO2024001669A1 (en) Antibodies targeting itga2 and antibody-drug conjugates comprising same
CA2696634A1 (en) Monoclonal antibodies against human acidic and basic ferritins and nucleotide and amino acid sequences thereof

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 08799192

Country of ref document: EP

Kind code of ref document: A2

WWE Wipo information: entry into national phase

Ref document number: 2698343

Country of ref document: CA

ENP Entry into the national phase

Ref document number: 2010524150

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 2008799192

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 12676348

Country of ref document: US