WO2017027422A1 - Constructs having a sirp-alpha domain or variant thereof - Google Patents

Constructs having a sirp-alpha domain or variant thereof Download PDF

Info

Publication number
WO2017027422A1
WO2017027422A1 PCT/US2016/045914 US2016045914W WO2017027422A1 WO 2017027422 A1 WO2017027422 A1 WO 2017027422A1 US 2016045914 W US2016045914 W US 2016045914W WO 2017027422 A1 WO2017027422 A1 WO 2017027422A1
Authority
WO
WIPO (PCT)
Prior art keywords
polypeptide
sirp
variant
domain
antibody
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/US2016/045914
Other languages
English (en)
French (fr)
Inventor
Jaume Pons
Laura Deming
Corey Goodman
Bang Janet Sim
Steven Elliot KAUDER
Hong Wan
Tracy Chia-Chien Kuo
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
ALX Oncology Inc
Original Assignee
Alexo Therapeutics Inc
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
Priority to IL294679A priority Critical patent/IL294679B2/en
Priority to DK16835723.4T priority patent/DK3331902T3/da
Priority to SI201631278T priority patent/SI3331902T1/sl
Priority to LTEP16835723.4T priority patent/LT3331902T/lt
Priority to NZ738950A priority patent/NZ738950A/en
Priority to SM20210424T priority patent/SMT202100424T1/it
Priority to MX2018001428A priority patent/MX395275B/es
Priority to CN202311337928.4A priority patent/CN117777267A/zh
Priority to PL16835723T priority patent/PL3331902T3/pl
Priority to CN202410176664.7A priority patent/CN117964732A/zh
Priority to EP21163641.0A priority patent/EP3913051A1/en
Priority to IL302491A priority patent/IL302491A/en
Priority to HRP20211167TT priority patent/HRP20211167T1/hr
Priority to JP2018506281A priority patent/JP6898303B2/ja
Priority to IL256989A priority patent/IL256989B/en
Priority to KR1020187006473A priority patent/KR102855748B1/ko
Priority to UAA201802321A priority patent/UA125638C2/uk
Priority to CN201680057835.8A priority patent/CN108350048B/zh
Priority to AU2016304794A priority patent/AU2016304794B2/en
Application filed by Alexo Therapeutics Inc filed Critical Alexo Therapeutics Inc
Priority to HK18116580.2A priority patent/HK1257372B/zh
Priority to EP16835723.4A priority patent/EP3331902B1/en
Priority to BR112018001353-6A priority patent/BR112018001353B1/pt
Priority to RU2018108103A priority patent/RU2740672C2/ru
Priority to KR1020257028619A priority patent/KR20250133806A/ko
Priority to RS20210912A priority patent/RS62151B1/sr
Priority to ES16835723T priority patent/ES2881771T3/es
Priority to MYPI2018000181A priority patent/MY185014A/en
Priority to PH1/2018/500269A priority patent/PH12018500269B1/en
Priority to CA2993835A priority patent/CA2993835A1/en
Publication of WO2017027422A1 publication Critical patent/WO2017027422A1/en
Anticipated expiration legal-status Critical
Priority to ZA2018/00857A priority patent/ZA201800857B/en
Priority to CONC2018/0002471A priority patent/CO2018002471A2/es
Priority to CY20211100674T priority patent/CY1124372T1/el
Priority to AU2021218004A priority patent/AU2021218004B2/en
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
    • A61K38/1774—Immunoglobulin superfamily (e.g. CD2, CD4, CD8, ICAM molecules, B7 molecules, Fc-receptors, MHC-molecules)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70596—Molecules with a "CD"-designation not provided for elsewhere
    • 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
    • A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
    • 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
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/6811—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a protein or peptide, e.g. transferrin or bleomycin
    • 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
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/06—Antiasthmatics
    • 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
    • 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
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A61P21/04—Drugs for disorders of the muscular or neuromuscular system for myasthenia gravis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/76—Albumins
    • C07K14/765—Serum albumin, e.g. HSA
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • 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
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • 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
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2818—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
    • 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
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2833—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against MHC-molecules, e.g. HLA-molecules
    • 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
    • C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
    • 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
    • C07K16/2887—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against CD20
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • 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
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • C07K14/4703—Inhibitors; Suppressors
    • 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
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C07K2317/524—CH2 domain
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C07K2317/526—CH3 domain
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/732—Antibody-dependent cellular cytotoxicity [ADCC]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Definitions

  • polypeptides comprising: a signal- regulatory protein a (SIRP-a) Dl variant comprising a SIRP-a Dl domain, or a fragment thereof, having an amino acid mutation at residue 80 relative to a wild-type SIRP-a Dl domain; and at least one additional amino acid mutation relative to a wild-type SIRP-a Dl domain at a residue selected from the group consisting of: residue 6, residue 27, residue 31, residue 47, residue 53, residue 54, residue 56, residue 66, and residue 92.
  • the wild type SIRP-a Dl domain has a sequence according to any one of SEQ ID NOs: 1-10.
  • the SIRP-a Dl domain comprises between one and nine additional amino acid mutations relative to a wild-type SIRP-a Dl domain at a residue selected from the group consisting of: residue 6, residue 27, residue 31, residue 47, residue 53, residue 54 residue 56, residue 66, and residue 92.
  • the SIRP-a Dl variant comprises the amino acid sequence
  • SIRP-a Dl variant has at least two amino acid substitutions relative to a wild-type SIRP-a Dl domain having a sequence according to SEQ ID NO: 1.
  • the SIRP-a Dl variant has at least two amino acid substitutions relative to a wild-type SIRP-a Dl domain having a sequence according to SEQ ID NO: 1.
  • EEELQXiIQPDKSVLVAAGETATLRCTX 2 TSLX 3 PVGPIQWFRGAGPGRX 4 LIYNQX 5 X 6 GX 7 F PRVTWSDXgTKimNMDFSIRIGXgXioXnXiiADAGTYYCX ⁇ KFRKGSPDDVEFKSGAGTEL SVRAKPS (SEQ ID NO: 218), wherein Xj is V, L, or I; X 2 is A, V, L, or I; X 3 is I, S, T, or F; X 4 is E, L, or V; X5 is K or R; X 6 is E or Q; X 7 is H, R, or P; X 8 is S,G, L, or T; X 9 is any amino acid; Xio is any amino acid; Xn is any amino acid; X 12 is any amino acid; and X13 is V or I; and wherein the SIRP-a Dl variant has at least two amino acid substitutions relative to a
  • X9 is A. In some embodiments, X9 is N. In some embodiments, X 10 is I. In some embodiments, X9 is N and X10 is P. In some embodiments, X9 is N and XI 1 is any amino acid other than S, T, or C. In some embodiments, X n is T. In some embodiments, X n is an amino acid other than T. In some embodiments, Xn is P. In some embodiments, X9 is N and Xn is any amino acid other than P. In some embodiments, the SIRP-a Dl variant comprises the amino acid sequence,
  • SIRP-a Dl variant has at least two amino acid substitutions relative to a wild-type SIRP-a Dl domain having a sequence according to SEQ ID NO: 219), wherein Xj is V, L, or I; X 2 is A, V, L, or I; X 3 is I, S, T, or F; X 4 is E, L, or V; X5 is K or R; X 6 is E or Q; X 7 is H, R, or P; X 8 is S, G, L, or T; X 9 is N; X 10 is any amino acid other than P; and Xn is V or I; and wherein the SIRP-a Dl variant has at least two amino acid substitutions relative to a wild-type SIRP-a Dl domain having a sequence according to SEQ ID NO: 219), wherein Xj is V, L, or I; X 2 is A, V, L, or I; X 3 is I, S, T, or F; X 4 is E, L, or
  • the SIRP-a Dl variant comprises the amino acid sequence, EEELQX 1 IQPDKSVLVAAGETATLRCTX 2 TSLX 3 PVGPIQWFRGAGPGRELIYNQX 4 EGX 5 FPR VTTVSDX 6 TKRNNMDFSIRIGX 7 ITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKPS (SEQ ID NO: 52), wherein Xj is V, L, or I; X 2 is A, I, or L; X 3 is I, T, S, or F; X 4 is K or R; X 5 is
  • the SIRP-a Dl variant has at least two amino acid substitutions relative to a wild-type SIRP-a Dl domain having a sequence according to SEQ ID NO: 1.
  • Xi is V or I
  • X 2 is A or I
  • X 3 is I or F
  • X 4 is K or R
  • X 5 is H or P
  • X6 is L or T
  • X7 is A.
  • the SIRP-a Dl variant has at least three amino acid substitutions relative to a wild-type SIRP-a Dl domain having a sequence according to SEQ ID NO: 1.
  • the SIRP-a Dl variant has at least four amino acid substitutions relative to a wild-type SIRP-a Dl domain having a sequence according to SEQ ID NO: 1. In some embodiments, the SIRP-a Dl variant has at least five amino acid substitutions relative to a wild- type SIRP-a Dl domain having a sequence according to SEQ ID NO: 1. In some embodiments, the SIRP-a Dl variant has at least six amino acid substitutions relative to a wild-type SIRP-a Dl domain having a sequence according to SEQ ID NO: 1. In some embodiments, the SIRP-a Dl variant has at least seven amino acid substitutions relative to a wild-type SIRP-a Dl domain having a sequence according to SEQ ID NO: 1. In some embodiments, Xi is I. In some embodiments, X 2 is
  • X3 is F.
  • X4 is R.
  • X5 is P.
  • Xe is T.
  • each of X 1; X 2 , X3, X 4 , X5, and Xe is not a wild-type amino acid.
  • the SIRP-a Dl variant has an amino acid sequence according to any one of SEQ ID NOs: 81-85. In some embodiments, the SIRP-a Dl variant comprises the amino acid sequence,
  • the polypeptide binds to human CD47 with a K D less than about 5 x 10 "9 M.
  • the polypeptide further comprises an Fc domain monomer linked to the N-terminus or the C-terminus of the polypeptide, wherein the Fc domain monomer is a human IgGl, IgG2, or IgG4 Fc region.
  • the Fc domain monomer comprises at least one mutation relative to a wild-type human IgGl, IgG2, or IgG4 Fc region.
  • the polypeptide has the amino acid sequence of any one of SEQ ID NO: 135, SEQ ID NO: 136, or SEQ ID NO: 137.
  • the Fc domain monomer comprises (a)_one of the following amino acid substitutions relative to wild type human IgGl: T366W, T366S, L368A, Y407V, T366Y, T394W, F405W, Y349T, Y349E, Y349V, L351T, L351H, L351N, L351K, P353S, S354D, D356K, D356R, D356S, E357K, E357R, E357Q, S364A, T366E, L368T, L368Y, L368E, K370E, K370D, K370Q, K392E, K392D, T394N, P395N, P396T, V397T, V397Q, L398T, D399K, D399R, D399N, F405T, F405H, F405R,
  • the Fc domain monomer comprises (a) one of the following amino acid substitutions relative to wild type human IgGl : T366W, T366S, L368A, Y407V, T366Y, T394W, F405W, Y349T, Y349E, Y349V, L351T, L351H, L351N, L351K, P353S, S354D, D356K, D356R, D356S, E357K, E357R, E357Q, S364A, T366E, L368T, L368Y, L368E, K370E, K370D, K370Q, K392E, K392D, T394N, P395N, P396T, V397T, V397Q, L398T, D399K, D399R, D399N, F405T, F405H, F405R,
  • the polypeptide exhibits a reduction of phagocytosis in a phagocytosis assay compared to a polypeptide with a wild-type human IgG Fc region.
  • the Fc domain monomer is linked to a second polypeptide comprising a second Fc domain monomer to form an Fc domain dimer.
  • the second Fc domain monomer is linked to an additional polypeptide.
  • the additional polypeptide comprises an antibody variable domain.
  • the antibody variable domain targets an antigen expressed on a cell.
  • the cell is a cancer cell.
  • the antibody variable domain targets a cell surface protein involved in immune cell regulation.
  • the additional polypeptide comprises a therapeutic protein.
  • the therapeutic protein is a cytokine, an interleukin, an antigen, a steroid, an anti-inflammatory agent, or an immunomodulatory agent.
  • the additional polypeptide comprises a SIRP-a Dl variant.
  • the polypeptide further comprises a human serum albumin (HSA) (SEQ ID NO: 12).
  • HSA human serum albumin
  • the HSA comprises a C34S or K573P amino acid substitution relative to SEQ ID NO: 12.
  • the polypeptide has an amino acid sequence according to any one of SEQ ID NOs: 152-159.
  • the polypeptide further comprises an albumin-binding peptide.
  • the albumin- binding peptide comprises the amino acid sequence DICLPRWGCLW (SEQ ID NO: 160).
  • the polypeptide further comprises a polyethylene glycol (PEG) polymer.
  • PEG polyethylene glycol
  • the PEG polymer is joined to a cysteine substitution in the polypeptide.
  • polypeptides comprising: a signal- regulatory protein a (SIRP-a) Dl variant, wherein the SIRP-a Dl variant comprises the amino acid sequence,
  • one of the Fc domain monomers in the Fc domain dimer comprises a human IgGl Fc region comprising L234A, L235A, G237A, and N297A mutations.
  • the polypeptide comprises an amino acid sequence according to any one of SEQ ID NOs: 98-104, 107-113, 116-122, or 135-137.
  • the Fc variant exhibits ablated or reduced binding to an Fey receptor compared to a wild-type version of a human IgG Fc region.
  • the IgGl or IgG2 Fc variant exhibits ablated or reduced binding to CD 16a, CD32a, CD32b, CD32c, and CD64 Fey receptors compared to a wild-type version of a human IgGl or IgG2 Fc region.
  • the IgG4 Fc variant exhibits ablated or reduced binding to CD 16a and CD32b Fey receptors compared to a wild-type version of the human IgG4 Fc region.
  • the IgGl or IgG2 Fc variant exhibits ablated or reduced binding to Clq compared to a wild-type version of a human IgGl or IgG2 Fc fusion.
  • the Fc variant binds to an Fey receptor with a KD greater than about 5 x 10 "6 M.
  • polypeptides comprising an Fc variant, wherein the Fc variant comprises an Fc domain dimer having two Fc domain monomers, wherein each Fc domain monomer independently is selected from (i) a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A; (ii) a human IgG2 Fc region consisting of mutations A330S, P331 S and N297A; or (iii) a human IgG4 Fc region comprising mutations S228P, E233P, F234V, L235A, delG236, and N297A.
  • At least one of the Fc domain monomers is a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A. In some embodiments, at least one of the Fc domain monomers is a human IgG2 Fc region consisting of mutations A330S, P33 IS and N297A. In some embodiments, the Fc variant exhibits ablated or reduced binding to an Fey receptor compared to the wild-type version of the human IgG Fc region.
  • the Fc variant exhibits ablated or reduced binding to CD16a, CD32a, CD32b, CD32c, and CD64 Fey receptors compared to the wild- type version of the human IgG Fc region. In some embodiments, the Fc variant exhibits ablated or reduced binding to Clq compared to the wild-type version of the human IgG Fc fusion. In some embodiments, at least one of the Fc domain monomers is a human IgG4 Fc region comprising mutations S228P, E233P, F234V, L235A, delG236, and N297A.
  • the Fc variant exhibits ablated or reduced binding to a Fey receptor compared to the wild-type human IgG4 Fc region. In some embodiments, the Fc variant exhibits ablated or reduced binding to CD 16a and CD32b Fey receptors compared to the wild-type version of its human IgG4 Fc region. In some embodiments, the Fc variant binds to an Fey receptor with a KD greater than about 5 x 10 "6 M. In some embodiments, the polypeptide further comprises a CD47 binding polypeptide. In some embodiments, the Fc variant exhibits ablated or reduced binding to an Fey receptor compared to a wild-type version of a human IgG Fc region.
  • the CD47 binding polypeptide does not cause acute anemia in rodents and non-human primates. In some embodiments, the CD47 binding polypeptide does not cause acute anemia in humans. In some embodiments, the CD47 binding polypeptide is a signal-regulatory protein a (SIRP-a) polypeptide or a fragment thereof. In some embodiments, the SIRP-a polypeptide comprises a SIRP-a Dl variant comprising the amino acid sequence,
  • EEELQX 1 IQPDKSVLVAAGETATLRCTX 2 TSLX 3 PVGPIQWFRGAGPGRX 4 LIYNQX 5 EGX 6 FP RVT SDX 7 TKPv MDFSIRIGX 8 ITPADAGTYYCX 9 KFRKGSPDDVEFKSGAGTELSVRAK PS (SEQ ID NO: 51), wherein Xi is V or I; X 2 is A or I; X 3 is I or F; X 4 is E or V; X 5 is K or R; X 6 is H or P; X 7 is L or T; X 8 is any amino acid other than N; and X9 is V or I.
  • the SIRP-a polypeptide comprises a SIRP-a Dl variant wherein Xi is V or I; X 2 is A or I; X 3 is I or F; X 4 is E; X 5 is K or R; X 6 is H or P; X 7 is L or T; X 8 is not N; and X9 is V.
  • polypeptides comprising: a signal- regulatory protein a (SIRP-a) Dl variant, wherein the SIRP-a Dl variant is a non-naturally occurring high affinity SIRP-a Dl domain, wherein the SIRP-a Dl variant binds to human CD47 with an affinity that is at least 10-fold greater than the affinity of a naturally occurring Dl domain; and an Fc domain monomer, wherein the Fc domain monomer is linked to a second polypeptide comprising a second Fc domain monomer to form an Fc domain, wherein the Fc domain has ablated or reduced effector function.
  • the non-naturally occurring high affinity SIRP-a Dl domain comprises an amino acid mutation at residue 80.
  • polypeptides comprising a signal- regulatory protein a (SIRP-a) Dl variant, wherein the SIRP-a Dl variant binds CD47 from a first species with a K D less than 250 nM; and wherein the SIRP-a Dl variant binds CD47 from a second species with a K D less than 250 nM; and the K D for CD47 from the first species and the K D for CD47 from the second species are within 100 fold of each other; wherein the first species and the second species are selected from the group consisting of: human, rodent, and non-human primate.
  • the SIRP-a Dl variant binds CD47 from at least 3 different species.
  • the non-human primate is cynomolgus monkey.
  • polypeptides comprising: (a) a signal- regulatory protein a (SIRP-a) Dl domain that binds human CD47 with a KD less than 250 nM; and (b) an Fc domain monomer linked to the N-terminus or the C-terminus of the SIRP-a Dl domain, wherein the polypeptide does not cause acute anemia in rodents and non-human primates.
  • the polypeptide is a non-naturally occurring variant of a human SIRP-a.
  • administration of the polypeptide in vivo results in hemoglobin reduction by less than 50% during the first week after administration.
  • the polypeptide further comprises at least one Fc variant, wherein the Fc variant is selected from (i) a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A; (ii) a human IgG2 Fc region consisting of mutations A330S, P33 IS and N297A; or (iii) a human IgG4 Fc region comprising mutations S228P, E233P, F234V, L235A, delG236, and N297A.
  • the Fc variant is a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A. In some embodiments, the Fc variant is a human IgG2 Fc region consisting of mutations A330S, P33 IS and N297A. In some embodiments, the Fc variant is a human IgG4 Fc region comprising mutations S228P, E233P, F234V, L235A, delG236, and N297A.
  • the polypeptide comprises a signal-regulatory protein a (SIRP-a) Dl variant comprising a SIRP-a Dl domain, or a fragment thereof, having an amino acid mutation at residue 80 relative to a wild-type SIRP-a Dl domain; and at least one additional amino acid mutation relative to a wild-type SIRP-a Dl domain at a residue selected from the group consisting of: residue 6, residue 27, residue 31, residue 47, residue 53, residue 54, residue 56, residue 66, and residue 92.
  • SIRP-a signal-regulatory protein a
  • the polypeptide comprises a signal-regulatory protein a (SIRP-a) Dl variant, wherein the SIRP-a Dl variant comprises the amino acid sequence, EEXiX 2 QX 3 lQPDKX 4 VX 5 VAAGEX 6 X 7 X 8 LX 9 CTXioTSLX tillPVGPIQWFRGAGPXi 2 PvXi 3 LIYN QXi 4 Xi 5 GXi 6 FPRVTrVSXnXi 8 TXi 9 RX 2 oNMDFX 2 iIX 22 lX 23 X 24 lTX 25 ADAGTYYCX 26 KX 27 K GSPDX 28 X 29 EX 30 KSGAGTELSVRX 31 KPS (SEQ ID NO: 47), wherein Xj is E, or G; X 2 is L, I, or V; X3 is V, L, or I; X 4 is S, or F; X 5 is L, or S; Xe is S,
  • the polypeptide comprises an Fc variant, wherein the Fc variant comprises an Fc domain dimer having two Fc domain monomers, wherein each Fc domain monomer independently is selected from (i) a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A; (ii) a human IgG2 Fc region consisting of mutations A330S, P33 I S and N297A; or (iii) a human IgG4 Fc region comprising mutations S228P, E233P, F234V, L235A, delG236, and N297A.
  • the polypeptide comprises a signal- regulatory protein a (SIRP-a) Dl variant, wherein the SIRP-a Dl variant is a non-naturally occurring high affinity SIRP-a Dl domain, wherein the SIRP-a Dl variant binds to human CD47 with an affinity that is at least 10-fold greater than the affinity of a naturally occurring Dl domain; and an Fc domain monomer, wherein the Fc domain monomer is linked to a second polypeptide comprising a second Fc domain monomer to form an Fc domain, wherein the Fc domain has ablated or reduced effector function.
  • the non-naturally occurring high affinity SIRP-a Dl domain comprises an amino acid mutation at residue 80.
  • the polypeptide comprises a signal-regulatory protein a (SIRP-a) Dl variant, wherein the SIRP-a D 1 variant binds CD47 from a first species with a K D less than 250 nM; and wherein the SIRP-a D l variant binds CD47 from a second species with a KD less than 250 nM; and the KD for CD47 from the first species and the K D for CD47 from the second species are within 100 fold of each other; wherein the first species and the second species are selected from the group consisting of: human, rodent, and non-human primate.
  • SIRP-a D 1 variant binds CD47 from a first species with a K D less than 250 nM
  • the SIRP-a D l variant binds CD47 from a second species with a KD less than 250 nM
  • the KD for CD47 from the first species and the K D for CD47 from the second species are within 100 fold of each other; wherein the first species and the second species
  • the polypeptide comprises (a) a signal-regulatory protein a (SIRP-a) Dl domain that binds human CD47 with a K D less than 250 nM; and (b) an Fc domain monomer linked to the N-terminus or the C-terminus of the SIRP-a Dl domain, wherein the polypeptide does not cause acute anemia in rodents and non-human primates.
  • the disease or disorder is a cancer, an autoimmune disease, or an inflammatory disease.
  • the disease or disorder is a cancer
  • the cancer is selected from solid tumor cancer, hematological cancer, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, non-Hodgkin lymphoma, Hodgkin lymphoma, multiple myeloma, bladder cancer, pancreatic cancer, cervical cancer, endometrial cancer, lung cancer, bronchus cancer, liver cancer, ovarian cancer, colon and rectal cancer, stomach cancer, gastric cancer, gallbladder cancer, gastrointestinal stromal tumor cancer, thyroid cancer, head and neck cancer, oropharyngeal cancer, esophageal cancer, melanoma, non-melanoma skin cancer, Merkel cell carcinoma, virally induced cancer, neuroblastoma, breast cancer, prostate cancer, renal cancer, renal cell cancer, renal pelvis cancer, leukemia, lymphoma, sarcoma, glioma, brain tumor, and carcinoma.
  • the disease or disorder is an autoimmune disease or an inflammatory disease
  • the autoimmune disease or the inflammatory disease is selected from multiple sclerosis, rheumatoid arthritis, a spondyloarthropathy, systemic lupus erythematosus, an antibody-mediated inflammatory or autoimmune disease, graft versus host disease, sepsis, diabetes, psoriasis, atherosclerosis, Sjogren's syndrome, progressive systemic sclerosis, scleroderma, acute coronary syndrome, ischemic reperfusion, Crohn's Disease, endometriosis, glomerulonephritis, myasthenia gravis, idiopathic pulmonary fibrosis, asthma, acute respiratory distress syndrome (ARDS), vasculitis, and inflammatory autoimmune myositis.
  • multiple sclerosis rheumatoid arthritis, a spondyloarthropathy, systemic lupus erythematosus
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the method further comprises administering at least one additional agent.
  • the at least one additional agent is an antibody, tumor associated antigen, or a non-antibody therapeutic.
  • at least two additional agents are administered.
  • the at least two additional agents comprise two antibodies.
  • the at least two additional agents comprise an antibody and a tumor associated antigen.
  • the at least one additional agent is an antibody.
  • the antibody is a human IgGl isotype antibody.
  • the antibody is a human IgG2 isotype antibody. In some embodiments, the antibody is a human IgG4 isotype antibody. In some embodiments, the antibody is selected from an anti-HER2 antibody, anti-CD20 antibody, anti-CD19 antibody, anti-CS l antibody, anti-CD38 antibody, anti-EGFR antibody, anti-PDl antibody, anti-OX40 antibody, anti- PD-1 antibody, anti-PD-Ll antibody, anti-CD274 antibody, anti-CTLA-4 antibody, anti-CD 137 antibody, anti-4-lBB antibody, anti-B7-H3 antibody, anti-FZD7 antibody, anti-CD27 antibody, anti-CCR4 antibody, anti-CD38 antibody, anti-CSFlR antibody, anti-CSF antibody, anti-CD30 antibody, anti-BAFF antibody, anti-VEGF antibody, or anti-VEGFR2 antibody.
  • the antibody is selected from an anti-HER2 antibody, anti-CD20 antibody, anti- CD 19 antibody, anti-CSl antibody, anti-CD38 antibody, anti-PD-1 antibody, anti-RANKL antibody, or anti-PD-Ll antibody.
  • the at least one additional agent is at least one antibody and the antibody is selected from cetuximab, necitumumab, pembrolizumab, nivolumab, pidilizumab, MEDI0680, MED 16469, atezolizumab, avelumab, durvalumab,
  • the antibody is trastuzumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is rituximab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-1 13, 116-122, 135-137, or 152-159.
  • the antibody is cetuximab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152- 159.
  • the antibody is daratumumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is belimumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • n the antibody is bevacizumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is denosumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is pantimumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-1 13, 116-122, 135-137, or 152-159.
  • the antibody is ramucirumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152- 159.
  • the antibody is necitumumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is nivolumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is pembrolizumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is avelumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is atezolizumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is durvalumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is durvalumab.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is MEDI0680.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is pidilizumab.
  • the SIRP-a D l variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107-113, 116-122, 135-137, or 152-159.
  • the antibody is BMS-93659.
  • the SIRP-a Dl variant has a sequence according to any one of SEQ ID NOs: 78-85, 98-104, 107- 113, 116-122, 135-137, or 152-159.
  • the at least one additional agent is a tumor associated antigen and the tumor associated antigen elicits an immune response.
  • the at least one additional agent is an antibody and the antibody targets a
  • the antibody targets a HLA/peptide or MHC/peptide complex comprising NY-ESO-1/LAGE1, SSX-2, MAGE family (MAGE- A3), gpl00/pmell7, Melan-A/MART- 1 , gp75/TRPl, tyrosinase, TRP2, CEA, PSA, TAG- 72, Immature laminin receptor, MOK RAGE-1, WT-1, Her2/neu, EphA3, SAP-1, BING-4, Ep- CAM, MUCl, PRAME, survivin, Mesothelin, BRCAl/2 (mutated), CDK4, CML66, MART-2, p53 (mutated), Ras (mutated), ⁇ -catenin (mutated), TGF-PRII (mutated), HPV E6, or E7.
  • the antibody is ESK1, RL1B, Pr20, or 3.2G1.
  • FIG. 2 is an illustration of a SIRP-a construct including a SIRP-a Dl domain or variant thereof joined to a first Fc domain monomer and an antibody variable domain joined to a second Fc domain monomer, wherein the first Fc domain monomer and the second Fc domain monomer combine to form an Fc domain;
  • FIG. 3 is an illustration of a SIRP-a construct including a SIRP-a Dl domain or variant thereof joined to a first Fc domain monomer and a therapeutic protein joined to a second Fc domain monomer, wherein the first Fc domain monomer and the second Fc domain monomer combine to form an Fc domain;
  • FIG. 4A is an illustration of a SIRP-a construct including a SIRP-a Dl domain or variant thereof joined to a first Fc domain monomer having a knob mutation, which forms an Fc domain with a second Fc domain monomer having a hole mutation
  • FIG. 4B is an illustration of a SIRP-a construct including a SIRP-a Dl domain or variant thereof joined to a first Fc domain monomer having a hole mutation, which forms an Fc domain with a second Fc domain monomer having a knob mutation;
  • FIG. 5A is an illustration of a SIRP-a construct including a SIRP-a Dl domain or variant thereof joined to an Fc domain monomer
  • FIG. 5B is an illustration of a SIRP-a construct which is a homodimer of the construct illustrated in FIG. 5A;
  • FIG. 6 exemplifies SPR binding data for monofunctional and bifunctional SIRP-a constructs including a SIRP-a Dl domain;
  • FIG. 7 exemplifies phagocytosis of DLD- 1 -GFP-Luciferase tumor cells by human monocyte-derived macrophages in the presence of varying concentrations of SIRP-a polypeptide constructs;
  • FIG. 8 exemplifies phagocytosis of DLD- 1 -GFP-Luciferase tumor cells by human monocyte-derived macrophages in the presence of varying concentrations of SIRP-a polypeptide constructs;
  • FIG. 9 exemplifies phagocytosis of DLD- 1 -GFP-Luciferase tumor cells by human monocyte-derived macrophages in the presence of varying concentrations of SIRP-a polypeptide constructs;
  • FIG. 10 exemplifies half-life stability of SIRP-a polypeptides over a defined time period
  • FIG. 11 exemplifies hemagglutination assay data for SIRP-a polypeptide constructs
  • FIG. 12 exemplifies survival curves of mice syngeneic tumor models treated with
  • FIG. 13 exemplifies a tumor volume analysis of mice syngeneic tumor models treated with SIRP-a polypeptide constructs in combination with anti-mPD-Ll;
  • FIG. 14 exemplifies binding of various concentrations of Clq complement to SIRP- ⁇ - Fc fusions
  • FIG. 15 exemplifies phagocytosis of MM1R cells by human monocyte-derived macrophages in the presence of varying concentrations of SIRP-a polypeptide constructs
  • FIG. 16 exemplifies phagocytosis of MM1R cells by human monocyte-derived macrophages in the presence of varying concentrations of SIRP-a polypeptide constructs
  • FIG. 17 exemplifies phagocytosis of N87 cells by human monocyte-derived macrophages in the presence of varying concentrations of SIRP-a polypeptide constructs
  • FIG. 18 exemplifies molecular weight analysis of a SIRP-a Dl variant having a
  • FIG. 19A exemplifies tumor growth of human GFP-Luc-Raji lymphoma cells in a
  • FIG. 19B exemplifies a scatter plot of tumor volume of the tumors described in FIG. 19A
  • FIG. 19C exemplifies hemoglobin values of the treated mice described in FIG. 19A
  • FIG. 20 exemplifies red blood cell counts taken from mice treated with either a
  • SIRP-a wildtype IgGl Fc construct or a SIRP-a IgGl Fc variant construct are SIRP-a wildtype IgGl Fc construct or a SIRP-a IgGl Fc variant construct.
  • “about” can mean a range of up to 20%, up to 10%, up to 5%, or up to 1% of a given value.
  • the term can mean within an order of magnitude, preferably within 5 -fold, and more preferably within 2-fold, of a value.
  • antibody refers to intact antibodies; antibody fragments, provided that they exhibit the desired biological activity (e.g. epitope binding); monoclonal antibodies; polyclonal antibodies; monospecific antibodies; multi-specific antibodies (e.g., bispecific antibodies); and antibody-like proteins.
  • antibody variable domain refers to the portions of the light and heavy chains of an antibody that include amino acid sequences of complementary determining regions (CDRs, e.g., CDR LI, CDR L2, CDR L3, CDR HI, CDR H2, and CDR H3) and framework regions (FRs).
  • CDRs complementary determining regions
  • FRs framework regions
  • linker refers to a linkage between two elements, e.g., protein domains.
  • a linker can be a covalent bond or a spacer.
  • spacer refers to a moiety (e.g., a polyethylene glycol (PEG) polymer) or an amino acid sequence (e.g., a 1-200 amino acid sequence) occurring between two polypeptides or polypeptide domains to provide space or flexibility (or both space and flexibility) between the two polypeptides or polypeptide domains.
  • an amino acid spacer is part of the primary sequence of a polypeptide (e.g., joined to the spaced polypeptides or polypeptide domains via the polypeptide backbone).
  • the term "therapeutically effective amount” refers to an amount of a polypeptide or a pharmaceutical composition containing a polypeptide described herein, e.g., a polypeptide having a SIRP-a Dl domain or variant thereof, that is sufficient and effective in achieving a desired therapeutic effect in treating a patient having a disease, such as a cancer, e.g., solid tumor or hematological cancer.
  • a therapeutically effective amount of polypeptide will avoid adverse side effects.
  • the term "pharmaceutical composition” refers to a medicinal or pharmaceutical formulation that includes an active ingredient as well as excipients or diluents (or both excipients and diluents) and enables the active ingredient to be administered by suitable methods of administration.
  • the pharmaceutical compositions disclosed herein include pharmaceutically acceptable components that are compatible with the polypeptide.
  • the pharmaceutical composition is in tablet or capsule form for oral administration or in aqueous form for intravenous or subcutaneous administration, for example by injection.
  • the terms "subject,” “individual,” and “patient” are used interchangeably to refer to a vertebrate, for example, a mammal. Mammals include, but are not limited to, murines, simians, humans, farm animals, sport animals, and pets. Tissues, cells, and their progeny of a biological entity obtained in vivo or cultured in vitro are also encompassed. None of the terms entail supervision of a medical professional.
  • binding affinity refers to the strength of the binding interaction between two molecules.
  • binding affinity refers to the strength of the sum total of non-covalent interactions between a molecule and its binding partner, such as a high affinity SIRP-a Dl variant and CD47.
  • binding affinity refers to intrinsic binding affinity, which reflects a 1 : 1 interaction between members of a binding pair.
  • the binding affinity between two molecules is commonly described by the dissociation constant (KD) or the association constant (KA). TWO molecules that have low binding affinity for each other generally bind slowly, tend to dissociate easily, and exhibit a large K D .
  • K D Two molecules that have high affinity for each other generally bind readily, tend to remain bound longer, and exhibit a small K D .
  • the K D of two interacting molecules is determined using known methods and techniques, e.g., surface plasmon resonance (SPR). KD can be calculated as the ratio of k 0ff /k on .
  • KD less than refers to a numerically smaller KD value and an increasing binding affinity relative to the recited K D value.
  • K D greater than refers to a numerically larger K D value and a decreasing binding affinity relative to the recited K D value.
  • acute anemia refers to a decrease of red blood cell mass or hemoglobin of 30% during the first five days after administration of a compound or treatment.
  • SIRP-a Signal-Regulatory Protein a
  • polypeptides comprising a signal- regulatory protein a (SIRP-a) Dl variant comprising a SIRP-a Dl domain, or a fragment thereof, having an amino acid mutation at residue 80 relative to a wild-type SIRP-a Dl domain; and at least one additional amino acid mutation relative to a wild-type SIRP-a Dl domain at a residue selected from the group consisting of: residue 6, residue 27, residue 31, residue 47, residue 53, residue 54, residue 56, residue 66, and residue 92.
  • SIRP-a signal- regulatory protein a
  • polypeptides comprising an Fc variant, wherein the Fc variant comprises an Fc domain dimer having two Fc domain monomers, wherein each Fc domain monomer independently is selected from (i) a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A; (ii) a human IgG2 Fc region consisting of mutations A330S, P331 S and N297A; or (iii) a human IgG4 Fc region comprising mutations S228P, E233P, F234V, L235A, delG236, and N297A.
  • SIRP-a Signal-regulatory protein a
  • SIRP-alpha Signal-regulatory protein a
  • CD47 a protein broadly expressed on many cell types in the body.
  • the interaction of SIRP-a with CD47 prevents engulfment of "self cells, which can otherwise be recognized by the immune system. It has been observed that high CD47 expression on tumor cells can act, in acute myeloid leukemia and several solid tumor cancers, as a negative prognostic factor for survival.
  • SIRP-a comprises 3 highly homologous immunoglobulin (Ig)-like extracellular domains— Dl, D2, and D3.
  • the SIRP-a Dl domain (“Dl domain”) refers to the membrane distal, extracellular domain of SIRP-a and mediates binding of SIRP-a to CD47.
  • SIRP-a polypeptide refers to any SIRP-a polypeptide or fragment thereof that is capable of binding to CD47.
  • Table 1 shows the amino acid sequences of the Dl domains of the ten naturally occurring wild-type human SIRP-a Dl domain variants (SEQ ID NOs: 1-10).
  • a SIRP-a polypeptide comprises a SIRP-a Dl domain. In some embodiments, a SIRP-a polypeptide comprises a wild-type Dl domain, such as those provided in SEQ ID NOs: 1-10. In some
  • a SIRP-a polypeptide includes a D2 or D3 domain (or both a D2 and a D3 domain) (Table 3) of a wild-type human SIRP-a.
  • KGSXisXieDXnEFKSGAGTELSVRXigKPS Xi is E or G; X2 is S or F; X3 is L or S; X 4 is T or S; X5
  • high affinity SIRP-a D 1 variant refers to a polypeptide comprising a SIRP-a Dl domain or a CD47-binding portion of a SIRP-a polypeptide that has a higher affinity to CD47 than wild-type SIRP-a.
  • a high affinity SIRP-a Dl variant comprises at least one amino acid substitution, deletion, or insertion (or a combination thereof) relative to a wild-type SIRP-a.
  • high affinity SIRP-a Dl variants disclosed herein comprise a
  • a high affinity SIRP-a Dl variant comprises one or more amino acid substitutions, insertions, additions, or deletions relative to a wild-type D l domain shown in SEQ ID NOs: 1-10.
  • Table 2 lists exemplary amino acid substitutions in each SIRP-a Dl domain variant (SEQ ID NOs: 13-22).
  • the SIRP-a Dl domain polypeptide or high affinity SIRP-a Dl variant comprises a fragment of the Dl domain.
  • the SIRP-a polypeptide fragment or high affinity SIRP-a variant fragment comprises an amino acid sequence of less than 10 amino acids in length, about 10 amino acids in length, about 20 amino acids in length, about 30 amino acids in length, about 40 amino acids in length, about 50 amino acids in length, about 60 amino acids in length, about 70 amino acids in length, about 80 amino acids in length, about 90 amino acids in length, about 100 amino acids in length, or more than about 100 amino acids in length.
  • the SIRP-a D 1 domain fragments retain the ability to bind to CD47.
  • a polypeptide of the disclosure comprising a high affinity
  • SIRP-a Dl variant binds with higher binding affinity to CD47 than a wild-type human SIRP-a Dl domain.
  • the high affinity SIRP-a Dl variant binds to human CD47 with at least 1-fold (e.g., at least 1.5-fold, 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 5-fold or greater than 5- fold) affinity than the affinity of a naturally occurring Dl domain.
  • the high affinity SIRP-a Dl variant binds to human CD47 with at least 1-fold (e.g., at least 10-fold, 100- fold, 1000-fold or greater than 1000-fold) affinity than the affinity of a naturally occurring Dl domain.
  • the term “optimized affinity” or “optimized binding affinity” refers to an optimized strength of the binding interaction between a polypeptide disclosed herein, including a high affinity SIRP-a Dl variant, and CD47.
  • the polypeptide binds primarily or with higher affinity to CD47 on cancer cells and does not substantially bind or binds with lower affinity to CD47 on non-cancer cells.
  • the binding affinity between the polypeptide and CD47 is optimized such that the interaction does not cause clinically relevant toxicity or decreases toxicity compared to a variant which binds with maximal affinity.
  • the polypeptide including a high affinity SIRP-a Dl variant in order to achieve an optimized binding affinity between a polypeptide provided herein and CD47, the polypeptide including a high affinity SIRP-a Dl variant is developed to have a lower binding affinity to CD47 than which is maximally achievable.
  • the high affinity SIRP-a variants disclosed herein cross react with rodent, non-human primate (NHP), and human CD47.
  • immunogenicity refers to the property of a protein (e.g., a therapeutic protein) which causes an immune response in the host as though it is a foreign antigen.
  • the immunogenicity of a protein can be assayed in vitro in a variety of different ways, such as through in vitro T-cell proliferation assays.
  • minimal immunogenicity refers to an immunogenicity of a protein (e.g., a therapeutic protein) that has been modified, e.g., through amino acid substitutions, to be lower (e.g., at least 10%, 25%, 50%, or 100% lower) than the immunogenicity before the amino acid substitutions are introduced (e.g., an unmodified protein).
  • a protein e.g., a therapeutic protein
  • a protein is modified to have minimal immunogenicity and causes no or very little host immune response even though it is a foreign antigen.
  • the high affinity SIRP-a Dl variant has minimal immunogenicity.
  • a SIRP-a polypeptide of the disclosure administered to a subject has the same amino acid sequence as that of the SIRP-a polypeptide in a biological sample of the subject, except for amino acid changes which increase affinity of the SIRP-a Dl variant.
  • the polypeptide variants disclosed herein lower the risk of side effects compared to anti-CD47 antibodies or wild-type SIRP-a.
  • the polypeptide variants disclosed herein lower the risk of anemia compared to anti-CD47 antibodies or wild-type SIRP-a.
  • the polypeptide variants disclosed herein do not cause acute anemia in rodent or non-human primates (NHP) studies.
  • Table 2 lists specific amino acid substitutions in a high affinity SIRP-a Dl variant relative to each Dl domain sequence.
  • a high affinity SIRP-a Dl variant includes one or more (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen or more) of the substitutions listed in Table 2.
  • a high affinity SIRP-a Dl variant includes at most fourteen amino acid substitutions relative to a wild-type Dl domain.
  • a high affinity SIRP-a Dl variant includes at most ten amino acid substitutions relative to a wild-type Dl domain.
  • a high affinity SIRP-a includes at most ten amino acid substitutions relative to a wild-type Dl domain.
  • D l variant includes at most seven amino acid substitutions relative to a wild-type Dl domain.
  • a high affinity SIRP-a Dl variant of the disclosure has at least 90% (e.g., at least 92%, 95%, 97% or greater than 97%) amino acid sequence identity to a sequence of a wild- type Dl domain.
  • a high affinity SIRP-a Dl variant is a chimeric high affinity
  • SIRP-a Dl variant that includes a portion of two or more wild-type Dl domains or variants thereof
  • a chimeric high affinity SIRP-a Dl variant includes at least two portions (e.g., three, four, five or more portions) of wild-type Dl domains or variants thereof, wherein each of the portions is from a different wild-type Dl domain.
  • a chimeric high affinity SIRP-a Dl variant further includes one or more amino acid substitutions listed in Table 2.
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of: EEEXiQX 2 IQPDKSVLVAAGETX3TLRCTX4TSLX 5 PVGPIQWFRGAGPGRX6LiYNQX 7 X 8 GX 9 FPRVTTVSDXioTXnPvNNMDFSIRISNITPADAGTYYCXi 2 KXi 3 RKGSPDDVEXi 4 KSGAGTEL SVRAKPS (SEQ ID NO: 21), wherein Xj is L, I, or V; X 2 is V, L, or, I; X 3 is A or V; X 4 is A, I, or L; X 5 is I, T, S, or F; X 6 is E, V, or L; X 7 is K or R; X 8 is E or Q; X 9 is H, P, or R; Xi 0 is L, T, or G; Xii is K or R; X
  • a polypeptide includes a high affinity
  • SIRP-a Dl variant having a sequence of any one of SEQ ID NOs: 13, 16-18, and 21, wherein Xi is
  • X 2 is V, L, or, I. In any of the
  • X3 is A or V.
  • X4 is A, I, or L.
  • X5 is I, T, S, or F.
  • Xe is E, V, or L.
  • X7 is K or R.
  • X 8 is E or Q.
  • X9 is H, P, or R.
  • X 10 is L, T, or G. In any of the
  • Xn is K or R.
  • X 12 is V or I.
  • X13 is F, L, V.
  • X14 is F or V.
  • the polypeptide of this aspect of the disclosure includes no more than six amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1, 4-6, and 9.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1, 4-6, and 9. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1, 4-6, and 9. In some embodiments, the polypeptide binds CD47 with at least 1000- fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1, 4-6, and 9.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a KD less than 1 x 10-8 M, less than 5 x 10-9 M, less than 1 x 10-9 M, less 5 x 10-10 M, less than 1 x 10-10 M or less than 1 x 10-11 M.
  • a SIRP-a D l variant polypeptide or fragment thereof binds to CD47 with a KD between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • the polypeptide has the sequence of any one of SEQ ID NOs: 14, 15, 19, and 22, wherein Xi is L, I, or V.
  • X 2 is V, L, or, I.
  • X 3 is A or V.
  • X 4 is V, I, or L.
  • X5 is I, T, S, or F.
  • X6 is E, V, or L.
  • X7 is K or R.
  • Xg is E or Q.
  • X9 is H, P, or R.
  • X 10 is S, T, or G.
  • Xn is K or R.
  • X 12 is V or I.
  • X13 is F, L, or V.
  • X14 is F or V.
  • the polypeptide of this aspect of the disclosure includes no more than six amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 2, 3, 7, and 10.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 2, 3, 7, and 10. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 2, 3, 7, and 10. In some embodiments, the polypeptide binds CD47 with at least 1000-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 2, 3, 7, and 10.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D less than 1 x 10 "8 M, less than 5 x 10 "9 M, less than 1 x 10 "9 M, less 5 x 10 "10 M, less than 1 x 10 "10 M or less than 1 x 10 "n M.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • the polypeptide has the sequence of SEQ ID NO: 20, wherein
  • Xi is L, I, or V.
  • X 2 is V, L, or, I.
  • X3 is A or V.
  • X4 is A, I, or L.
  • X5 is I, T, S, or F.
  • Xe is E, V, or L.
  • X7 is K or R.
  • Xg is E or Q.
  • X9 is H, P, or R.
  • X 10 is S, T, or G.
  • Xn is K or R.
  • Xn is V or I.
  • X13 is F, L, or V.
  • X14 is F or V.
  • the polypeptide of this aspect of the disclosure includes no more than six amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of SEQ ID NO: 8.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of SEQ ID NO: 8. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of SEQ ID NO: 8. In some embodiments, the polypeptide binds CD47 with at least 1000-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of SEQ ID NO: 8.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D less than 1 x 10 "8 M, less than 5 x 10 "9 M, less than 1 x 10 "9 M, less 5 x 10 "10 M, less than 1 x 10 "10 M or less than 1 x 10 "n M.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • X 2 is L, I, or V.
  • X3 is V, L, or, I.
  • X4 is S or F.
  • X5 is L or S.
  • Xe is S or T.
  • X7 is A or V.
  • Xg is I or T.
  • X9 is H or R.
  • X 10 is A, V, I, or L.
  • Xn is I, T, S, or F.
  • Xn is A or G.
  • X13 is E, V, or L.
  • X 14 is K or R.
  • X15 is E or Q.
  • X e is H, P, or R. In any of the
  • Xn is D or E.
  • X $ is S, L, T, or G.
  • X19 is K or R.
  • X 2 o is E or D.
  • X 2 i is S or P.
  • X 22 is S or R.
  • X 2 3 is S or G.
  • X 2 4 is V or I.
  • X 3 ⁇ 4 is F, L, V.
  • X 3 ⁇ 4 is D or absent. In any of the aforementioned embodiments, ⁇ 2 ⁇ is T or V. In any of the aforementioned embodiments, X 3 ⁇ 4 is F or V. In any of the aforementioned embodiments, X 2 9 is A or G. In some embodiments, the polypeptide of this aspect of the disclosure includes no more than six amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1-10.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1- 10. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1- 10. In some embodiments, the polypeptide binds CD47 with at least 1000-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1-10.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D less than 1 x 10 "8 M, less than 5 x 10 "9 M, less than 1 x 10 "9 M, less 5 x 10 "10 M, less than 1 x 10 "10 M or less than 1 x 10 "n M.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a KD between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • a polypeptide of the disclosure including a high affinity
  • SIRP-a Dl variant further comprises a D2 domain having the sequence of SEQ ID NO: 24, a D3 domain having the sequence of SEQ ID NO: 25, or a D2 domain having the sequence of SEQ ID NO: 24 and a D3 domain having the sequence of SEQ ID NO: 25 of a wild-type human SIRP-a as shown in Table 3.
  • the high affinity SIRP-a Dl variant further comprises a fragment or variant of a D2 domain or a fragment or variant of a D3 domain.
  • the high affinity SIRP-a Dl variant further comprises a fragment or variant of a D2 domain and a fragment or variant of a D3 domain.
  • a high affinity SIRP-a D 1 variant is joined to a D2 or D3 domain by way of a linker.
  • a high affinity SIRP-a Dl variant is joined to a D2 and D3 domain by way of a linker.
  • a polypeptide of the disclosure including a high affinity
  • SIRP-a Dl variant is attached to an Fc domain monomer, a human serum albumin (HSA) or variant thereof, a serum-binding protein or peptide, or an organic molecule, e.g., a polymer (e.g., a PEG polymer), in order to improve the pharmacokinetic properties of the polypeptide, e.g., increase serum half-life.
  • a high affinity SIRP-a Dl variant is attached to an Fc domain monomer that is unable to dimerize.
  • Fc domain monomers, HSA proteins, serum-binding proteins or peptides, and organic molecules such as a PEG serve to increase the serum half-life of the polypeptides described herein.
  • a polypeptide of the disclosure including a high affinity SIRP-a Dl variant does not include the sequence of any one of SEQ ID NOs: 26-36 shown in Table 4. Table 4.
  • polypeptides and polypeptide constructs described herein are utilized in vitro for binding assays, such as immune assays.
  • the polypeptides and polypeptide constructs described herein are utilized in liquid phase or bound to a solid phase carrier.
  • polypeptides utilized for immunoassays are detectably labeled in various ways.
  • polypeptides and polypeptide constructs described herein are bound to various carriers and used to detect the presence of specific antigen expressing cells.
  • Examples of carriers include glass, polystyrene, polypropylene, polyethylene, dextran, nylon, amylases, natural and modified celluloses, polyacrylamides, agaroses, and magnetite.
  • the nature of the carrier can be either soluble or insoluble.
  • labels include enzymes, radioisotopes, fluorescent compounds, colloidal metals, chemilumine scent compounds, and bio-luminescent compounds.
  • Various techniques for binding labels to polypeptides disclosed herein are available.
  • the polypeptides are coupled to low molecular weight haptens. These haptens are then specifically detected by means of a second reaction.
  • the hapten biotin is used with avidin or the haptens dinitrophenol, pyridoxal, or fluorescein are detected with specific anti-hapten antibodies (e.g., anti-dinitrophenol antibodies, anti -pyridoxal antibodies, and anti-fluorescein antibodies respectively).
  • polypeptides comprising a signal- regulatory protein a (SIRP-a) Dl variant comprising a SIRP-a Dl domain, or a fragment thereof, having an amino acid mutation at residue 80 relative to a wild-type SIRP-a Dl domain; and at least one additional amino acid mutation relative to a wild-type SIRP-a Dl domain at a residue selected from the group consisting of: residue 6, residue 27, residue 31, residue 47, residue 53, residue 54, residue 56, residue 66, and residue 92.
  • SIRP-a signal- regulatory protein a
  • polypeptides comprising an Fc variant, wherein the Fc variant comprises an Fc domain dimer having two Fc domain monomers, wherein each Fc domain monomer independently is selected from (i) a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A; (ii) a human IgG2 Fc region consisting of mutations A330S, P331 S and N297A; or (iii) a human IgG4 Fc region comprising mutations S228P, E233P, F234V, L235A, delG236, and N297A.
  • a polypeptide in a composition disclosed herein comprises a high affinity SIRP-a Dl variant that has reduced or minimal glycosylation.
  • the Dl domain of each of the ten wild-type human SIRP-a proteins contains a single potential N-linked glycosylation site at amino acid N80 in the sequence N80ITP.
  • Expression of a SIRP-a Dl domain in Chinese Hamster Ovary (CHO) cells results in a major band of 16 kDa (non- glycosylated) and a minor band of higher molecular weight that was removed by Endo Hf.
  • Endo Hf is a recombinant protein fusion of Endoglycosidase H and maltose binding protein. Endo Hf cleaves within the chitobiose core of high mannose and some hybrid oligosaccharides from N- linked glycoproteins. This implies that a proline at amino acid position 83 can reduce the efficiency of glycosylation, leading to a protein with different degrees of glycosylation and therefore heterogeneity. For drug development, heterogeneity can give rise to challenges in process development. Therefore, to investigate the possibility of generating homogenous, non-glycosylated forms of high affinity SIPR-a Dl variants, in some embodiments, amino acid N80 of a SIPR-a Dl variant is mutated to Ala.
  • amino acid N80 in a high affinity SIRP-a Dl variant is replaced by any amino acid, including any naturally and non-naturally occurring amino acid, e.g., N80A and N80Q.
  • a high affinity SIRP-a Dl variant comprises an N80A mutation and at least 1 additional mutation (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional mutations or more).
  • the additional mutation is in the CD47 binding site.
  • the additional mutation is in the hydrophobic core of the Dl domain.
  • a polypeptide in a composition disclosed herein includes a high affinity SIRP-a Dl variant that has increased glycosylation relative to a wild-type SIRP-a Dl domain. Another option to increase homogeneity of the final product is to enhance the efficiency of glycosylation at amino acid N80 and generate high affinity SIRP-a Dl variants with increased glycosylation relative to a wild-type.
  • the amino acid P83 in the sequence NITP83 affects the degree of glycosylation at amino acid N80. In some embodiments, changing P83 to any amino acid increases the efficiency of glycosylation at N80.
  • amino acid P83 in a high affinity SIRP-a Dl variant is replaced by any amino acid, including naturally and non-naturally amino acids, e.g., P83V, P83A, P83I, and P83L.
  • a polypeptide of the disclosure is expressed in a cell that is optimized not to glycosylate proteins that are expressed by such cell, for example by genetic engineering of the cell line (e.g., genetically engineered yeast or mammalian host) or modifications of cell culture conditions such as addition of kifunensine or by using a naturally non-glycosylating host such as a prokaryote (E. coli, etc.).
  • Table 5 lists specific amino acid substitutions in a high affinity SIRP-a Dl variant relative to each Dl domain variant sequence.
  • a high affinity SIRP-a Dl variant includes one or more (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen or more) of the substitutions listed in Table 5.
  • the SIRP-a Dl variants are not glycosylated or are minimally glycosylated.
  • the SIRP-a D l variants are fully glycosylated or almost fully glycosylated.
  • a high affinity SIRP-a Dl variant includes at most fourteen amino acid substitutions relative to a wild-type Dl domain. In some embodiments, a high affinity SIRP-a Dl variant includes at most ten amino acid substitutions relative to a wild-type Dl domain. In some embodiments, a high affinity SIRP-a D l variant includes at most seven amino acid substitutions relative to a wild-type Dl domain. In some embodiments, a high affinity SIRP-a Dl variant of the disclosure has at least 90% (e.g., at least 92%, 95%, 97% or greater than 97%) amino acid sequence identity to a sequence of a wild- type Dl domain.
  • a high affinity SIRP-a Dl variant is a chimeric high affinity
  • SIRP-a Dl variant that includes a portion of two or more wild-type Dl domains or variants thereof
  • a chimeric high affinity SIRP-a Dl variant includes at least two portions (e.g., three, four, five or more portions) of wild-type Dl domains or variants thereof, wherein each of the portions is from a different wild-type Dl domain.
  • a chimeric high affinity SIRP-a Dl variant further includes one or more amino acid substitutions listed in Table 5.
  • R, S, T, V, W, Y; X 13 P, A, C, D, E, F, G, H, I, K, L, M, NO: 38
  • R, S, T, V, W, Y; X 13 P, A, C, D, E, F, G, H, I, K, L, M, NO: 39
  • R, S, T, V, W, Y; X 13 P, A, C, D, E, F, G, H, I, K, L, M, NO: 40
  • R, S, T, V, W, Y; X 13 P, A, C, D, E, F, G, H, I, K, L, M, NO: 41
  • R, S, T, V, W, Y; X 13 P, A, C, D, E, F, G, H, I, K, L, M, NO: 42
  • R, S, T, V, W, Y; X 13 P, A, C, D, E, F, G, H, I, K, L, M, NO: 43
  • R, S, T, V, W, Y; X 13 P, A, C, D, E, F, G, H, I, K, L, M, NO: 44
  • R, S, T, V, W, Y; X 13 P, A, C, D, E, F, G, H, I, K, L, M, NO: 45
  • a polypeptide includes a SIRV-ct D 1 variant having a sequence of:
  • X 8 is E or Q
  • X 9 is H, P, or R
  • X, 0 is L, T, or G
  • X, , is K or R
  • X, 2 is N, A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, or Y
  • X, 3 is P, A, C, D, E, F, G, H, i, K, L, M, N, Q, R, S, T, V, W, or Y
  • X, 4 is V or I
  • X, s is F, L, or V
  • « is F or V
  • die variant lias at least one amino acid substitution relative to a wild-type SI P-a Dl domain having the sequence of SEQ ID NO: 1.
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • eKSGAG TELSVRAKPS (SEQ ID NO: 40), wherein X, is L, I, or V; X 2 is V, L, or, I; X 3 is A or V; X ⁇ is A, 1, or L; X 5 is 1, T, S, or F; X 6 is E, V, or L; X 7 is K or R; X 8 is E or Q; X 9 is H, P, or R; ⁇ ⁇ 0 is L, T, or G; X trash is K or R; X I2 is N, A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, or Y; X, 3 is P, A, C, D, E, F, G, H, ⁇ , K, L, M, N, Q, R, S, T, V, W, or Y; X M is V or I; X theft is F, L, or V; and X l6 is
  • D l domain having the sequence of SEQ ID NO: 4.
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of any one of SEQ ID NOs: 37, 40-42, and 45, wherein Xi is L, I, or V.
  • X 2 is V, L, or, I.
  • X3 is A or V.
  • X4 is A, I, or L.
  • X5 is I, T, S, or F.
  • Xe is E, V, or L.
  • X7 is K or R.
  • Xg is E or Q.
  • X 9 is H, P, or R.
  • Xi 0 is L, T, or G.
  • Xn is K or R.
  • X 12 is N, A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, or Y.
  • X J3 is P, A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, or Y.
  • X14 is V or I.
  • X15 is F, L, V.
  • X 1 ⁇ 2 is F or V.
  • a polypeptide provided herein includes no more than ten amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1, 4-6, and 9. In some embodiments, the polypeptide provided herein includes no more than seven amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1, 4-6, and 9.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1, 4-6, and 9. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1, 4-6, and 9. In some embodiments, the polypeptide binds CD47 with at least 1000- fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1, 4-6, and 9.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D less than 1 x 10 "8 M, less than 5 x 10 "9 M, less than 1 x 10 "9 M, less 5 x 10 "10 M, less than 1 x 10 "10 M or less than 1 x 10 "n M.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • a polypeptide includes a SIRP-a Dl variant having a sequence of any one of SEQ ID NOs: 38, 39,
  • Xi is L, I, or V.
  • X2 is V, L, or, I.
  • X 3 is A or V.
  • X4 is V, I, or L.
  • X5 is I, T, S, or F.
  • Xe is E, V, or L.
  • X7 is K or R.
  • Xg is E or Q.
  • X9 is H, P, or R.
  • X10 is S, T, or G.
  • Xn is K or R.
  • X 12 is N, A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, or Y.
  • X J3 is P, A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, or Y.
  • X14 is V or I.
  • X15 is F, L, or V.
  • X 1 ⁇ 2 is F or V.
  • a polypeptide includes a SIRP-a Dl variant having no more than ten amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 2, 3, 1, and 10. In some embodiments, a polypeptide includes a SIRP-a Dl variant having no more than seven amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 2, 3, 7, and 10.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 2, 3, 7, and 10. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 2, 3, 7, and 10. In some embodiments, the polypeptide binds CD47 with at least 1000-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 2, 3, 7, and 10.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D less than 1 x 10 "8 M, less than 5 x 10 "9 M, less than 1 x 10 "9 M, less 5 x 10 "10 M, less than 1 x 10 "10 M or less than lxlO "11 M.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a KD between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • the polypeptide has the sequence of SEQ ID NO: 44, wherein Xi is L, I, or V.
  • X 2 is V, L, or, I.
  • X3 is A or V.
  • X 4 is A, I, or L.
  • X5 is I, T, S, or F.
  • Xe is E, V, or L.
  • X7 is K or R.
  • X 8 is E or Q. In any of the
  • X9 is H, P, or R.
  • X 10 is S, T, or G.
  • Xn is K or R.
  • X n is N, A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, or Y.
  • X J3 is P, A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, or Y.
  • Xi 4 is V or I.
  • any of the aforementioned embodiments is V or I.
  • X 15 is F, L, or V. In any of the aforementioned embodiments, X 1 ⁇ 2 is F or V.
  • a polypeptide includes a SIRP-a Dl variant having no more than ten amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of SEQ ID NO: 8. In some embodiments, a polypeptide includes a SIRP-a Dl variant having no more than seven amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of SEQ ID NO: 8.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of SEQ ID NO: 8. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of SEQ ID NO: 8. In some embodiments, the polypeptide binds CD47 with at least 1000-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of SEQ ID NO: 8.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D less than 1 x 10 "8 M, less than 5 x 10 "9 M, less than 1 x 10 "9 M, less 5 x 10 "10 M, less than 1 x 10 "10 M or less than 1 x 10 "n M.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • the disclosure features a polypeptide including a SIRP-a Dl variant having a sequence of:
  • the polypeptide has the sequence of SEQ ID NO: 47, wherein Xi is E or G.
  • X 2 is L, I, or V.
  • X 3 is V, L, or, I.
  • X 4 is S or F.
  • X5 is L or S.
  • Xe is S or T.
  • X7 is A or V.
  • Xg is I or T.
  • X 9 is H, R, or L.
  • X 10 is A, V, I, or L.
  • Xn is I, T, S, or F.
  • Xn is A or G.
  • X 13 is E, V, or L.
  • X J4 is K or R.
  • X i5 is E or Q.
  • X e is H, P, or R. In any of the
  • Xn is D or E.
  • Xi 8 is S, L, T, or G.
  • X19 is K or R.
  • X 20 is E or N.
  • X 21 is S or P.
  • X 22 is S or R.
  • X 23 is S or G.
  • X 24 is N, A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, or Y.
  • X 25 is P, A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, or Y.
  • X 2 6 is V or I.
  • X 27 is F, L, V.
  • X 28 is D or absent.
  • X 29 is T or V.
  • X30 is F or V.
  • X31 is A or G.
  • the polypeptide of this aspect of the disclosure includes no more than ten amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1-10. In some embodiments, the polypeptide of this aspect of the disclosure includes no more than seven amino acid substitutions relative to the wild-type SIRP- ⁇ Dl domain having the sequence of any one of SEQ ID NOs: 1-10.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1-10. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1-10. In some embodiments, the polypeptide binds CD47 with at least 1000-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1-10.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D less than 1 x 10 "8 M, less than 5 x 10 "9 M, less than 1 x 10 "9 M, less 5 x 10 "10 M, less than 1 x 10 "10 M or less than 1 x 10 "n M.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a KD between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • a polypeptide includes a SIRP-a Dl variant having a sequence of:
  • the disclosure features a polypeptide including a SIRP-a Dl variant having a sequence of: EEELQXiIQPDKSVLVAAGETATLRCTX 2 TSLX 3 PVGPIQWFRGAGPGRX 4 LIYNQX 5 X 6 GX 7 F PRVTWSDX 8 TKRNNMDFSIRIGX 9 ITPADAGTYYCXioKFRKGSPDDVEFKSGAGTELSVRA KPS (SEQ ID NO: 49), wherein Xj is V, L, or I; X 2 is A, I, V, or L; X 3 is I, F, S, or T; X 4 is E, V, or L; X 5 is K or R; X 6 is E or Q; X 7 is H, P, or R; X 8 is L, T, S, or G; X9 is A; and X 10 is V or I; and wherein the variant has at least one amino acid substitution relative to a wild-type SIRP-a D
  • the polypeptide has the sequence of SEQ ID NO: 49, wherein Xi is V, L or I.
  • X 2 is A, I, V, or L.
  • X3 is I, F, S, or T. In any of the
  • X4 is E, V, or L.
  • X5 is K or R.
  • Xe is E or Q.
  • X 7 is H, P, or R.
  • X 8 is L, T, S or G.
  • X9 is A.
  • X 1 0 is V or I.
  • the polypeptide has a high affinity SIRP-a Dl domain having at least 85% sequence identity (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) to SEQ ID NO: 49, wherein each of X 2, X3, X4, X5, ⁇ , X 7 , Xs, Xs>, and X10 are not a wild-type amino acid.
  • the polypeptide of this aspect of the disclosure includes no more than ten amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 1. In some embodiments, the polypeptide of this aspect of the disclosure includes no more than seven amino acid substitutions relative to the wild-type SIRP-a
  • D 1 domain having the sequence of any one of SEQ ID NO: 1.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 1. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 1. In some embodiments, the polypeptide binds CD47 with at least 1000-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 1.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a KD less than 1 x 10 "8 M, less than 5 x 10 "9 M, less than 1 x 10 "9 M, less 5 x 10 "10 M, less than 1 x 10 "10 M or less than 1 x 10 "n M.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • the disclosure features a polypeptide including a SIRP-a Dl variant having a sequence of:
  • the polypeptide has the sequence of SEQ ID NO: 50, wherein Xi is V or I.
  • X 2 is V or I.
  • X3 is I or F.
  • X4 is E or V.
  • X5 is K or R.
  • Xe is E or Q.
  • X7 is H or P.
  • X 8 is S or R.
  • X 9 is N or A.
  • X lo is V or I.
  • the polypeptide has a high affinity SIRP-a Dl domain having at least 85% sequence identity (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) to SEQ ID NO: 50, wherein each of X X 2j X3, Xzt, ⁇ 3 ⁇ 4, ⁇ , X 7 , ⁇ , X ⁇ >, and X lo is not a wild-type amino acid.
  • sequence identity e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity
  • the polypeptide of this aspect of the disclosure includes no more than ten amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 2. In some embodiments, the polypeptide of this aspect of the disclosure includes no more than seven amino acid substitutions relative to the wild-type SIRP-a
  • D 1 domain having the sequence of any one of SEQ ID NO: 2.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 2. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 2. In some embodiments, the polypeptide binds CD47 with at least 1000-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 2.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a KD less than 1 x 10 "8 M, less than 5 x 10 "9 M, less than 1 x 10 "9 M, less 5 x 10 "10 M, less than 1 x 10 "10 M or less than 1 x 10 "n M.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • the disclosure features a polypeptide including a SIRP-a Dl variant having a sequence of:
  • EEELQXiIQPDKSVLVAAGETATLRCTX 2 TSLX 3 PVGPIQWFRGAGPGRX 4 LIYNQX 5 EGX 6 FP RVT SDX 7 TKRN MDFSIRIGX 8 ITPADAGTYYCX 9 KFRKGSPDDVEFKSGAGTELSVRAK PS (SEQ ID NO: 51), wherein Xj is V or I; X 2 is A or I; X 3 is I or F; X 4 is E or V; X 5 is K or R; X 6 is H or P; X 7 is L or T; X 8 is any amino acid other than N; and X9 is V or I; and wherein the variant has at least one amino acid substitution relative to a wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 1.
  • the polypeptide has the sequence of SEQ ID NO: 5 1, wherein Xi is V or I.
  • X 2 is A or I.
  • X3 is I or F.
  • X 4 is E or V.
  • X5 is K or R.
  • Xe is H or P.
  • X7 is L or T.
  • X 8 is N or A.
  • X9 is V or I.
  • X 4 is not V.
  • the polypeptide has the sequence of SEQ ID NO: 5 1, wherein X 8 is A.
  • X 8 is A and Xi is V or I.
  • X 8 is A and X 2 is A or I.
  • X 8 is A and X3 is I or F.
  • X 8 is A and X 4 is E or V. In some embodiments, X 4 is not V.
  • X 8 is A and X5 is K or R.
  • X 8 is A and Xe is H or P. In any of the aforementioned embodiments, X 8 is A and X7 is A or V. In any of the aforementioned embodiments, X 8 is A and X 9 is V or I.
  • the polypeptide has the sequence of SEQ ID NO: 5 1, wherein X 8 is A.
  • X 8 is A and Xi is I.
  • X 8 is A and X 2 is I.
  • X 8 is A and X 3 is F.
  • X 8 is A and X 4 is V.
  • X 8 is A and X5 is R.
  • is A and Xe is P.
  • Xs is A and X7 is T.
  • Xs is A and X9 is I.
  • the polypeptide has a high affinity SIRP-a Dl domain having at least 85% sequence identity (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) to SEQ ID NO: 51, wherein each of Xj , X 2j X 3, X 4, ⁇ 3 ⁇ 4, ⁇ , X 7, Xs , and X 9 is not a wild-type amino acid.
  • sequence identity e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity
  • the polypeptide of this aspect of the disclosure includes no more than ten amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 1. In some embodiments, the polypeptide of this aspect of the disclosure includes no more than seven amino acid substitutions relative to the wild-type SIRP-a
  • D 1 domain having the sequence of any one of SEQ ID NO: 1.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 1. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NOs: 1. In some embodiments, the polypeptide binds CD47 with at least 1000-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 1.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D less than 1 x 10 "8 M, less than 5 x 10 "9 M, less than 1 x 10 "9 M, less 5 x 10 "10 M, less than 1 x 10 "10 M or less than 1 x 10 "n M.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • the disclosure features a polypeptide including a SIRP-a Dl variant having a sequence of:
  • the polypeptide has the sequence of SEQ ID NO: 52, wherein Xi is V, L, or I.
  • X 2 is A, I, or L.
  • X 3 is I, T, S, or F.
  • X 4 is K or R.
  • X5 is H or P.
  • Xe is L, T, or G.
  • X 7 is N or A.
  • the polypeptide has the sequence of SEQ ID NO: 52, wherein Xi is V or I.
  • X 2 is A or I.
  • X 3 is I or F.
  • X 4 is K or R.
  • X 5 is H or P.
  • Xe is L or T.
  • X 7 is N or A.
  • the polypeptide has the sequence of SEQ ID NO: 52, wherein X 7 is A.
  • X 7 is A and Xi is V or I.
  • X7 is A and X 2 is A or I.
  • X 7 is A and X 3 is I or F.
  • X 7 is A and X 4 is K or R. In any of the aforementioned
  • X 7 is A and X5 is H or P. In any of the aforementioned embodiments, X 7 is A and Xe is L or T.
  • the polypeptide has the sequence of SEQ ID NO: 52, wherein X 7 is A.
  • X 7 is A and Xi is I.
  • X7 is A and X 2 is I.
  • X 7 is A and X 3 is F.
  • X 7 is A and X 4 is R.
  • X 7 is A and X5 is P.
  • X 7 is A and Xe is T.
  • the polypeptide has a high affinity SIRP-a Dl domain having at least 85% sequence identity (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) to SEQ ID NO: 52, wherein each of Xj , X 2, X3 , X 4, ⁇ 3 ⁇ 4 , ⁇ ⁇ , and X7 is not a wild-type amino acid.
  • the polypeptide of this aspect of the disclosure includes no more than ten amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 1. In some embodiments, the polypeptide of this aspect of the disclosure includes no more than seven amino acid substitutions relative to the wild-type SIRP-a
  • D 1 domain having the sequence of any one of SEQ ID NO: 1.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 1. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 1. In some embodiments, the polypeptide binds CD47 with at least 1000-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 1.
  • fragments include polypeptides of less than 10 amino acids in length, about 10 amino acids in length, about 20 amino acids in length, about 30 amino acids in length, about 40 amino acids in length, about 50 amino acids in length, about 60 amino acids in length, about 70 amino acids in length, about 80 amino acids in length, about 90 amino acids in length, about 100 amino acids in length, or more than about 100 amino acids in length. Fragments retain the ability to bind to CD47.
  • SIRP-a Dl variant polypeptides and fragments thereof bind to CD47 with a higher affinity than a SIRP-a polypeptide binds to CD47. For example, in some
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D less than 1 x 10 "8 M, less than 5 x 10 "9 M, less than 1 x 10 "9 M, less 5 x 10 "10 M, less than 1 x 10 "10 M or less than 1 x 10 "n M.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • the disclosure features a polypeptide including a SIRP-a Dl variant having a sequence of:
  • the polypeptide has the sequence of SEQ ID NO: 212, wherein Xi is V, L, or I.
  • X 2 is V, I, or L.
  • X 3 is I, T, S, or F.
  • X4 is K or R.
  • X5 is H or P.
  • Xe is S, T, or G.
  • X7 is A.
  • the polypeptide has the sequence of SEQ ID NO: 212, wherein Xi is V or I.
  • X 2 is V or I.
  • X 3 is I or F.
  • X4 is K or R.
  • X 5 is H or P.
  • Xe is S or T.
  • X7 is A.
  • the polypeptide has the sequence of SEQ ID NO: 212, wherein X 7 is A.
  • X7 is A and Xi is V or I.
  • X7 is A and X 2 is V or I.
  • X 7 is A and X 3 is I or F.
  • X7 is A and X4 is K or R. In any of the aforementioned
  • X7 is A and X5 is H or P. In any of the aforementioned embodiments, X7 is A and Xe is S or T.
  • the polypeptide has the sequence of SEQ ID NO: 212, wherein X 7 is A.
  • X7 is A and Xi is I.
  • X7 is A and X2 is I.
  • X7 is A and X3 is F. In any of the
  • X7 is A and X4 is R. In any of the aforementioned embodiments, X7 is A and X5 is P. In any of the aforementioned embodiments, X7 is A and Xe is T.
  • the polypeptide has a high affinity SIRP-a Dl domain having at least 85% sequence identity (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) to SEQ ID NO: 212, wherein each of X X2, X3, X4, ⁇ 3 ⁇ 4 , ⁇ ⁇ , and X7 is not a wild-type amino acid.
  • the polypeptide of this aspect of the disclosure includes no more than ten amino acid substitutions relative to the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 2. In some embodiments, the polypeptide of this aspect of the disclosure includes no more than seven amino acid substitutions relative to the wild-type SIRP-a
  • D 1 domain having the sequence of any one of SEQ ID NO: 2.
  • the polypeptide binds CD47 with at least 10-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 2. In some embodiments, the polypeptide binds CD47 with at least 100-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 2. In some embodiments, the polypeptide binds CD47 with at least 1000-fold greater binding affinity than the wild-type SIRP-a Dl domain having the sequence of any one of SEQ ID NO: 2.
  • fragments include polypeptides of less than 10 amino acids in length, about 10 amino acids in length, about 20 amino acids in length, about 30 amino acids in length, about 40 amino acids in length, about 50 amino acids in length, about 60 amino acids in length, about 70 amino acids in length, about 80 amino acids in length, about 90 amino acids in length, about 100 amino acids in length, or more than about 100 amino acids in length. Fragments retain the ability to bind to CD47.
  • SIRP-a Dl variant polypeptides and fragments thereof bind to CD47 with a higher affinity than a SIRP-a polypeptide binds to CD47. For example, in some
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D less than 1 x 10 "8 M, less than 5 x 10 "9 M, less than 1 x 10 "9 M, less 5 x 10 "10 M, less than 1 x 10 "10 M or less than 1 x 10 "n M.
  • a SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a K D between about 500 nM and 100 nM, between about 100 nM and 50 nM, between about 50 nM and 10 nM, between about 10 nM and 5 nM, between about 5 nM and 1 nM, between about 1 nM and 500 pM, between about 500 pM and 100 pM, between about 100 pM and 50 pM, or between about 50 pM and 10 pM.
  • Described herein, in some embodiments, is a polypeptide comprising a SIRP-a Dl variant having a sequence according to:
  • EEELQX 1 IQPDKSVLVAAGETATLRCTX 2 TSLX 3 PVGPIQWFRGAGPGRX 4 LIYNQX 5 X 6 GX 7 F PRVTWSDX 8 TKRNNMDFSIRIGX 9 XioXnXi 2 ADAGTYYCXi 3 KFRKGSPDDVEFKSGAGTEL SVRAKPS (SEQ ID NO: 218), wherein Xj is V, L, or I; X 2 is A, V, L, or I; X 3 is I, S, T, or F; X 4 is E, L, or V; X5 is K or R; X 6 is E or Q; X 7 is H, R, or P; X 8 is S,G, L, or T; X 9 is any amino acid; X 10 is any amino acid; Xn is any amino acid; X 12 is any amino acid; and X13 is V or I; and wherein the SIRP-a Dl variant has at least two amino acid substitutions relative
  • the polypeptide has the sequence of SEQ ID NO: 212, X 9 is A. In any of the aforementioned embodiments in this aspect of the disclosure, X 9 is N. In any of the aforementioned embodiments in this aspect of the disclosure X 1 0 is I. In any of the
  • X 9 is N and X10 is P.
  • XI 1 is any amino acid other than S, T, or C.
  • Xn is T.
  • Xn is an amino acid other than T.
  • Xn is P.
  • X 9 is N and Xn is any amino acid other than P.
  • Described herein, in some embodiments, is a polypeptide comprising a SIRP-a Dl variant having a sequence according to:
  • SIRP-a Dl variant has at least two amino acid substitutions relative to a wild-type SIRP-a Dl domain having a sequence according to SEQ ID NO: 219), wherein Xj is V, L, or I; X 2 is A, V, L, or I; X 3 is I, S, T, or F; X 4 is E, L, or V; X5 is K or R; X 6 is E or Q; X 7 is H, R, or P; X 8 is S, G, L, or T; X 9 is N; X 10 is any amino acid other than P; and Xn is V or I; and wherein the SIRP-a Dl variant has at least two amino acid substitutions relative to a wild-type SIRP-a Dl domain having a sequence according to SEQ ID NO: 219), wherein Xj is V, L, or I; X 2 is A, V, L, or I; X 3 is I, S, T, or F; X 4 is E, L, or
  • compositions which include a SIRP-a Dl variant polypeptide having the amino acid sequence of SEQ ID NO: 48, or a fragment thereof.
  • the SIRP-a Dl variant polypeptide or fragment thereof binds to CD47 with a higher affinity compared to the affinity that a SIRP-a polypeptide binds to the CD47.
  • the SIRP-a Dl variant polypeptide binds to CD47 with a KD less than 1 x 10 "8 M, or less than 1 x 10 "9 M, less than 1 x 10 "10 M or less than 1 x 10 "n M.
  • the above-mentioned SIRP-a Dl variant polypeptides are attached or fused to a second polypeptide.
  • the second polypeptide includes, without limitation, an Fc polypeptide, an Fc variant, an HSA polypeptide, an albumin peptide, a PEG polymer or a fragment of the foregoing.
  • a SIRP-a Dl variant polypeptide is selected from any one of SEQ ID NOs: 53-87 and 213 shown in Table 6.
  • VEFKSGAGTELSVRAKPS EEELQIIQPDKSVLVAAGETATLRCTITSLFPVGPIQWFRGAGPGRELIYNQ
  • VEFKSGAGTELSVRAKPS EEELQIIQPDKSVLVAAGETATLRCTITSLFPVGPIQWFRGAGPGRELIYNQ
  • the polypeptide includes a high affinity SIRP-a Dl domain that has at least 85% sequence identity (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) to any variant provided in Table 6.
  • the polypeptide includes a high affinity SIRP-a Dl domain that has at least 85% sequence identity (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) to SEQ ID NOs: 80, 81, or 85 in Table 6.
  • polypeptides comprising a signal- regulatory protein a (SIRP-a) Dl variant comprising a SIRP-a Dl domain, or a fragment thereof, having an amino acid mutation at residue 80 relative to a wild-type SIRP-a Dl domain; and at least one additional amino acid mutation relative to a wild-type SIRP-a Dl domain at a residue selected from the group consisting of: residue 6, residue 27, residue 31, residue 47, residue 53, residue 54, residue 56, residue 66, and residue 92.
  • SIRP-a signal- regulatory protein a
  • polypeptides comprising an Fc variant, wherein the Fc variant comprises an Fc domain dimer having two Fc domain monomers, wherein each Fc domain monomer independently is selected from (i) a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A; (ii) a human IgG2 Fc region consisting of mutations A330S, P331 S and N297A; or (iii) a human IgG4 Fc region comprising mutations S228P, E233P, F234V, L235A, delG236, and N297A.
  • Antibodies that target cell surface antigens can trigger immunostimulatory and effector functions that are associated with Fc receptor (FcR) engagement on immune cells.
  • Fc receptor Fc receptor
  • Binding of the Fc region to Fc receptors on cell surfaces can trigger a number of biological responses including phagocytosis of antibody-coated particles (antibody-dependent cell-mediated phagocytosis, or ADCP), clearance of immune complexes, lysis of antibody-coated cells by killer cells (antibody- dependent cell-mediated cytotoxicity, or ADCC) and, release of inflammatory mediators, placental transfer, and control of immunoglobulin production. Additionally, binding of the CI component of complement to antibodies can activate the complement system. Activation of complement can be important for the lysis of cellular pathogens. However, the activation of complement can also stimulate the inflammatory response and can also be involved in autoimmune hypersensitivity or other immunological disorders. Variant Fc regions with reduced or ablated ability to bind certain Fc receptors are useful for developing therapeutic antibodies and Fc -fusion polypeptide constructs which act by targeting, activating, or neutralizing ligand functions while not damaging or destroying local cells or tissues.
  • a SIRP-a Dl polypeptide construct comprises a non-naturally occurring high affinity SIRP-a Dl variant linked to an Fc domain monomer which forms an Fc domain having ablated or reduced effector function.
  • a Fc domain monomer refers to a polypeptide chain that includes second and third antibody constant domains (e.g., CH2 and CH3).
  • an Fc domain monomer also includes a hinge domain.
  • the Fc domain monomer is of any immunoglobulin antibody isotype, including IgG, IgE, IgM, IgA, and IgD.
  • an Fc domain monomer is of any IgG subtype (e.g., IgGl, IgG2, IgG2a, IgG2b, IgG2c, IgG3, and IgG4).
  • Fc domain monomers include as many as ten changes from a wild-type Fc domain monomer sequence (e.g., 1-10, 1-8, 1-6, 1-4 amino acid substitutions, additions or insertions, deletions, or combinations thereof) that alter the interaction between an Fc domain and an Fc receptor.
  • a wild-type Fc domain monomer sequence e.g., 1-10, 1-8, 1-6, 1-4 amino acid substitutions, additions or insertions, deletions, or combinations thereof
  • Fc domain refers to a dimer of two Fc domain monomers.
  • two Fc domain monomers dimerize by the interaction between the two CH3 antibody constant domains, as well as one or more disulfide bonds that form between the hinge domains of the two dimerized Fc domain monomers.
  • an Fc domain is mutated to lack effector functions, for example a "dead Fc domain.”
  • each of the Fc domain monomers in an Fc domain includes amino acid substitutions in the CH2 antibody constant domain to reduce the interaction or binding between the Fc domain and an Fc receptor, such as an Fey receptor (FcyR), an Fca receptor (FcaR), or an Fee (FceR).
  • FcyR Fey receptor
  • FcaR Fca receptor
  • Fee Fee
  • a high affinity SIRP-a Dl variant e.g., any of the variants described in Tables 2, 5, and 6 is fused to an Fc domain monomer of an immunoglobulin or a fragment of an Fc domain monomer.
  • an Fc domain monomer of an immunoglobulin or a fragment of an Fc domain monomer is capable of forming an Fc domain with another Fc domain monomer.
  • an Fc domain monomer or a fragment of an Fc domain monomer is not capable of forming an Fc domain with another Fc domain monomer.
  • an Fc domain monomer or a fragment of an Fc domain is fused to a polypeptide of the disclosure to increase serum half-life of the polypeptide.
  • an Fc domain monomer or a fragment of an Fc domain monomer fused to a polypeptide of the disclosure dimerizes with a second Fc domain monomer to form an Fc domain which binds an Fc receptor, or alternatively, an Fc domain monomer binds to an Fc receptor.
  • an Fc domain or a fragment of the Fc domain fused to a polypeptide to increase serum half-life of the polypeptide does not induce any immune system- related response.
  • a SIRP-a polypeptide or construct provided herein includes a SIRP-a Dl domain or variant thereof joined to a first Fc domain monomer and an antibody variable domain joined to a second Fc domain monomer, in which the first and second Fc domain monomers combine to form an Fc domain (e.g., a heterodimeric Fc domain).
  • An Fc domain is the protein structure that is found at the C-terminus of an immunoglobulin.
  • An Fc domain includes two Fc domain monomers that are dimerized by the interaction between the CH3 antibody constant domains.
  • a wild-type Fc domain forms the minimum structure that binds to an Fc receptor, e.g., FcyRI, FcyRIIa, FcyRIIb, FcyRIIIa, FcyRIIIb, and FcyRTV.
  • the Fc domain is not involved directly in binding an antibody to its target, but can be involved in various effector functions, such as participation of the antibody in antibody- dependent cellular toxicity.
  • the Fc domain in a SIRP-a polypeptide or construct of the disclosure comprise amino acid substitutions, additions or insertions, deletions, or any combinations thereof that lead to decreased effector function such as decreased antibody- dependent cell-mediated cytotoxicity (ADCC), decreased complement-dependent cytolysis (CDC), decreased antibody-dependent cell-mediated phagocytosis (ADCP), or any combinations thereof.
  • the SIRP-a polypeptides or constructs of the disclosure are characterized by decreased binding (e.g., minimal binding or absence of binding) to a human Fc receptor and decreased binding (e.g., minimal binding or absence of binding) to complement protein Clq.
  • the SIRP-a constructs of the disclosure are characterized by decreased binding (e.g., minimal binding or absence of binding) to human FcyRI, FcyRIIA, FcyRJIB, FcyRIIIB, FcyRIIIB, or any combinations thereof, and Clq.
  • the Fc domains in SIRP-a constructs of the disclosure are of the IgG class and comprise one or more amino acid substitutions at E233, L234, L235, G236, G237, D265, D270, N297, E318, K320, K322, A327, A330, P331, or P329 (numbering according to the EU index of Kabat (Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991))).
  • polypeptide constructs comprising a non-native Fc region described herein exhibit reduced or ablated binding to at least one of Fey receptors CD16a, CD32a, CD32b, CD32c, and CD64 as compared to a polypeptide construct comprising a native Fc region.
  • the polypeptide constructs described herein exhibit reduced or ablated binding to CD16a, CD32a, CD32b, CD32c, and CD64 Fey receptors.
  • CDC refers to a form of cytotoxicity in which the complement cascade is activated by the complement component Clq binding to antibody Fc.
  • polypeptide constructs comprising a non-native Fc region described herein exhibit at least a 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater reduction in Clq binding compared to a polypeptide construct comprising a wild-type Fc region.
  • polypeptide constructs comprising a non-native Fc region as described herein exhibit reduced CDC as compared to a polypeptide construct comprising a wild-type Fc region.
  • polypeptide constructs comprising a non-native Fc region as described herein exhibit at least a 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater reduction in CDC compared to a polypeptide construct comprising a wild-type Fc region.
  • polypeptide constructs comprising a non-natural Fc variant as described herein exhibit negligible CDC as compared to a polypeptide construct comprising a wild-type Fc region.
  • the Fc variants herein are minimally glycosylated or have reduced glycosylation relative to a wild-type sequence.
  • deglycosylation is accomplished with a mutation of N297A, or by mutating N297 to any amino acid which is not N.
  • the N- Xaal-Xaa2-Xaa3 motif refers to residues 297-300 as designated according to Kabat et al., 1991.
  • a mutation to any one or more of N, Xaal, Xaa2, or Xaa3 results in deglycosylation of the Fc variant.
  • variants of antibody IgG constant regions e.g., Fc variants possess a reduced capacity to specifically bind Fey receptors or have a reduced capacity to induce phagocytosis.
  • variants of antibody IgG constant regions possess a reduced capacity to specifically bind Fey receptors and have a reduced capacity to induce phagocytosis.
  • an Fc domain is mutated to lack effector functions, typical of a "dead" Fc domain.
  • an Fc domain includes specific amino acid substitutions that are known to minimize the interaction between the Fc domain and an Fey receptor.
  • an Fc domain monomer is from an IgGl antibody and includes one or more of amino acid substitutions L234A, L235A, G237A, and N297A (as designated according to the EU numbering system per Kabat et al, 1991).
  • one or more additional mutations are included in such IgGl Fc variant.
  • additional mutations for human IgGl Fc variants include E318A and K322A.
  • a human IgGl Fc variant has up to 12, 11, 10, 9, 8, 7, 6, 5 or 4 or fewer mutations in total as compared to wild-type human IgGl sequence.
  • one or more additional deletions are included in such IgGl Fc variant.
  • the C-terminal lysine of the Fc IgGl heavy chain constant region provided in SEQ ID NO: 88 in Table 7 is deleted, for example to increase the homogeneity of the polypeptide when the polypeptide is produced in bacterial or mammalian cells.
  • a human IgGl Fc variant has up to 12, 11, 10, 9, 8, 7, 6, 5 or 4 or fewer deletions in total as compared to wild-type human IgGl sequence.
  • a IgGl Fc variant has a sequence according to any one of SEQ ID NO: 135, SEQ ID NO: 136, or SEQ ID NO: 137.
  • an Fc domain monomer is from an IgG2 or IgG4 antibody and includes amino acid substitutions A330S, P33 IS, or both A330S and P33 IS.
  • the Fc variant comprises a human IgG2 Fc sequence comprising one or more of A330S, P331 S and N297A amino acid substitutions (as designated according to the EU numbering system per Kabat, et al. (1991). In some embodiments, one or more additional mutations are included in such IgG2 Fc variants.
  • Non-limiting examples of such additional mutations for human IgG2 Fc variant include V234A, G237A, P238S, V309L and H268A (as designated according to the EU numbering system per Kabat et al. (1991)).
  • a human IgG2 Fc variant has up to 12, 11, 10, 9, 8, 7, 6, 5, 4, 3 or fewer mutations in total as compared to wild-type human IgG2 sequence.
  • one or more additional deletions are included in such IgG2 Fc variant.
  • the C-terminal lysine of the Fc IgG2 heavy chain constant region provided in SEQ ID NO: 89 in Table 7 is deleted, for example to increase the homogeneity of the polypeptide when the polypeptide is produced in bacterial or mammalian cells.
  • a human IgG2 Fc variant has up to 12, 11, 10, 9, 8, 7, 6, 5 or 4 or fewer deletions in total as compared to wild -type human IgG2 sequence.
  • the Fc variant is an IgG4 Fc variant
  • such Fc variant comprises a S228P mutation (as designated according to Kabat, et al. (1991)).
  • a human IgG4 Fc variant has up to 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 mutation(s) in total as compared to wild-type human IgG4 sequence.
  • the Fc variant includes at least one of the mutations L234A, L235A, G237A or N297A of an IgGl Fc region or at least one of the mutations A330S, P33 IS or N297A of an IgG2 Fc region. In some embodiments, the Fc variant includes at least two of the mutations L234A, L235A, G237A or N297A of an IgGl Fc region or at least two of the mutations A330S, P331 S or N297A of an IgG2 Fc region.
  • the Fc variant includes at least three of the mutations L234A, L235A, G237A or N297A of an IgGl Fc region or consists of the mutations A330S, P33 IS and N297A of an IgG2 Fc region. In some embodiments, the Fc variant consists of the mutations L234A, L235A, G237A and N297A.
  • the Fc variant exhibits reduced binding to an Fc receptor of the subject compared to the wild-type human IgG Fc region. In some embodiments, the Fc variant exhibits ablated binding to an Fc receptor of the subject compared to the wild-type human IgG Fc region. In some embodiments, the Fc variant exhibits a reduction of phagocytosis compared to the wild-type human IgG Fc region. In some embodiments, the Fc variant exhibits ablated
  • SEQ ID NO: 88 and SEQ ID NO: 89 provide amino acid sequences of Fc IgGl and IgG2 heavy chain constant regions.
  • an Fc variant is any variant of SEQ ID NOs: 90-95 as shown in Table 7.
  • Antibody-dependent cell-mediated cytotoxicity which is also referred to herein as ADCC, refers to a form of cytotoxicity in which secreted Ig bound onto Fc receptors (FcRs) present on certain cytotoxic cells (e.g., Natural Killer (NK) cells and neutrophils) enabling these cytotoxic effector cells to bind specifically to an antigen-bearing target cell and subsequently kill the target cell.
  • FcRs Fc receptors
  • NK Natural Killer
  • Antibody-dependent cell-mediated phagocytosis which is also referred to herein as ADCP, refers to a form of cytotoxicity in which secreted Ig bound onto Fc receptors (FcRs) present on certain phagocytic cells (e.g., macrophages) enabling these phagocytic effector cells to bind specifically to an antigen-bearing target cell and subsequently engulf and digest the target cell.
  • FcRs Fc receptors
  • Ligand-specific high-affinity IgG antibodies directed to the surface of target cells can stimulate the cytotoxic or phagocytic cells and can be used for such killing.
  • polypeptide constructs comprising an Fc variant as described herein exhibit reduced ADCC or ADCP as compared to a polypeptide construct comprising a wild-type Fc region.
  • polypeptide constructs comprising an Fc variant as described herein exhibit at least a 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater reduction in ADCC or ADCP compared to a polypeptide construct comprising a wild-type Fc region.
  • polypeptide constructs comprising an Fc variant as described herein exhibit ablated ADCC or ADCP as compared to a polypeptide construct comprising a wild-type Fc region.
  • Complement-directed cytotoxicity which is also referred to herein as CDC, refers to a form of cytotoxicity in which the complement cascade is activated by the complement component Clq binding to antibody Fc.
  • polypeptide constructs comprising an Fc variant as described herein exhibit at least a 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater reduction in Clq binding compared to a polypeptide construct comprising a wild-type Fc region.
  • polypeptide constructs comprising an Fc variant as described herein exhibit reduced CDC as compared to a polypeptide construct comprising a wild-type Fc region.
  • polypeptide constructs comprising an Fc variant as described herein exhibit at least a 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater reduction in CDC compared to a polypeptide construct comprising a wild-type Fc region. In some cases, polypeptide constructs comprising an Fc variant as described herein exhibit negligible CDC as compared to a polypeptide construct comprising a wild-type Fc region.
  • Fc variants herein include those that exhibit reduced binding to an Fey receptor compared to the wild-type human IgG Fc region.
  • an Fc variant exhibits binding to an Fey receptor that is less than the binding exhibited by a wild-type human IgG Fc region to an Fey receptor, as described in the Examples.
  • an Fc variant has reduced binding to an Fey receptor by a factor of 10%, 20% 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% (fully ablated effector function).
  • the reduced binding is for any one or more Fey receptor, e.g., CD 16a, CD32a, CD32b, CD32c, or CD64.
  • the Fc variants disclosed herein exhibit a reduction of phagocytosis compared to its wild-type human IgG Fc region.
  • Such Fc variants exhibit a reduction in phagocytosis compared to its wild-type human IgG Fc region, wherein the reduction of phagocytosis activity is e.g., by a factor of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100%.
  • an Fc variant exhibits ablated phagocytosis compared to its wild-type human IgG Fc region.
  • the Fc variants disclosed herein are coupled to one or more fusion partners.
  • the fusion partner is a therapeutic moiety.
  • the fusion partner is selected to enable targeting of an expressed protein, purification, screening, display, and the like.
  • the fusion partner also affects the degree of binding to Fc receptors or the degree of phagocytosis reduction.
  • an Fc variant when coupled to a fusion partner, it forms a polypeptide construct as described below.
  • fusion partners are linked to the Fc variant sequence via a linker sequence .
  • the linker sequence generally comprises a small number of amino acids, such as less than ten amino acids, although longer linkers are also utilized.
  • the linker has a length less than 10, 9, 8, 7, 6, or 5 amino acids or shorter.
  • the linker has a length of at least 10, 1 1, 12, 13, 14, 15, 20, 25, 30, or 35 amino acids or longer.
  • a cleavable linker is employed.
  • a fusion partner is a targeting or signal sequence that directs an Fc variant protein and any associated fusion partners to a desired cellular location or to the extracellular media.
  • certain signaling sequences target a protein to be either secreted into the growth media, or into the periplasmic space, located between the inner and outer membrane of the cell.
  • a fusion partner is a sequence that encodes a peptide or protein that enables purification or screening.
  • Such fusion partners include, but are not limited to, polyhistidine tags (His-tags) (for example His6 and His 10) or other tags for use with
  • Immobilized Metal Affinity Chromatography (IMAC) systems e.g., Ni+2 affinity columns
  • GST fusions e.g., GST fusions
  • MBP fusions e.g., MBP fusions
  • Strep-tag e.g., MBP fusions
  • BSP biotinylation target sequence of the bacterial enzyme BirA e.g., BirA, and epitope tags which are targeted by antibodies (for example c-myc tags, flag-tags, and the like).
  • tags are useful for purification, for screening, or both.
  • an Fc variant is purified using a His-tag by immobilizing it to a Ni+2 affinity column, and then after purification the same His-tag is used to immobilize the antibody to a Ni+2 coated plate to perform an ELISA or other binding assay as described elsewhere herein.
  • a fusion partner enables the use of a selection method to screen Fc variants as described herein.
  • fusion partners that enable a variety of selection methods are available. For example, by fusing the members of an Fc variant library to the gene III protein, phage display can be employed. In some embodiments, fusion partners enable Fc variants to be labeled. Alternatively, in some embodiments, a fusion partner binds to a specific sequence on the expression vector, enabling the fusion partner and associated Fc variant to be linked covalently or noncovalently with the nucleic acid that encodes them.
  • the therapeutic moiety is, e.g., a peptide, a protein, an antibody, a siRNA, or a small molecule.
  • therapeutic antibodies that are coupled to the Fc variants of the present disclosure include, but are not limited to antibodies that recognize CD47.
  • therapeutic polypeptides that are coupled to the Fc variants of the present disclosure include, but are not limited to, CD47 binding polypeptides, including SIRP-a polypeptides. In such instances, the CD47 binding polypeptide is attached or fused to an Fc variant of the disclosure.
  • CD47 binding polypeptides include, but are not limited to, anti-CD47 antibodies or fragments thereof, and ligands of CD47 such as SIRP-a or a fragment thereof. Additional examples of CD47 binding polypeptides include, but are not limited to naturally-occurring forms of SIRP-a as well as mutants thereof.
  • a polypeptide comprising an Fc variant, wherein the Fc variant comprises an Fc domain dimer having two Fc domain monomers, wherein each Fc domain monomer independently is selected from (i) a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A; (ii) a human IgG2 Fc region consisting of mutations A330S, P33 IS and N297A; or (iii) a human IgG4 Fc region comprising mutations S228P, E233P, F234V, L235A, delG236, and N297A.
  • the Fc domain monomers are identical (i.e., homodimer). In some embodiments, the Fc domain monomers are different (i.e., heterodimer). In some embodiments, at least one of the Fc domain monomers in an Fc domain dimer is a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A. In some embodiments, at least one of the Fc domain monomers in an Fc domain dimer is a human IgG2 Fc region consisting of mutations A330S, P33 IS and N297A.
  • the Fc variant exhibits ablated or reduced binding to an Fey receptor compared to the wild-type version of the human IgG Fc region. In some embodiments, the Fc variant exhibits ablated or reduced binding to CD16a, CD32a, CD32b, CD32c, and CD64 Fey receptors compared to the wild- type version of the human IgG Fc region. In some embodiments, the Fc variant exhibits ablated or reduced binding to Clq compared to the wild-type version of the human IgG Fc fusion.
  • At least one of the Fc domain monomers in an Fc domain dimer is a human IgG4 Fc region comprising mutations S228P, E233P, F234V, L235A, delG236, and N297A.
  • the Fc variant exhibits ablated or reduced binding to a Fey receptor compared to the wild-type human IgG4 Fc region.
  • the Fc variant exhibits ablated or reduced binding to CD 16a and CD32b Fey receptors compared to the wild-type version of its human IgG4 Fc region.
  • the Fc variant binds to an Fey receptor with a KD greater than about 5 x 10-6 M.
  • the Fc variant further comprises a CD47 binding polypeptide.
  • the Fc variant exhibits ablated or reduced binding to an Fey receptor compared to a wild-type version of a human IgG Fc region.
  • the CD47 binding polypeptide does not cause acute anemia in rodents and non-human primates. In some embodiments, the CD47 binding polypeptide does not cause acute anemia in humans.
  • the CD47 binding polypeptide is a signal-regulatory protein a (SIRP-a) polypeptide or a fragment thereof.
  • SIRP-a polypeptide comprises a SIRP-a Dl variant comprising the amino acid sequence,
  • EEELQX 1 IQPDKSVLVAAGETATLRCTX 2 TSLX 3 PVGPIQWFRGAGPGRX 4 LIYNQX 5 EGX 6 FP RVT SDX 7 TKR MDFSIRIGX 8 ITPADAGTYYCX 9 KFRKGSPDDVEFKSGAGTELSVRAK PS (SEQ ID NO: 51), wherein Xj is V or I; X 2 is A or I; X 3 is I or F; X 4 is E or V; X 5 is K or R; X 6 is H or P; X 7 is L or T; X 8 is any amino acid other than N; and X9 is V or I.
  • the SIRP-a polypeptide comprises a SIRP-a Dl variant wherein Xi is V or I; X 2 is A or I; X 3 is I or F; X4 is E; X5 is K or R; Xe is H or P; X7 is L or T; xs is not N; and X9 is V.
  • a polypeptide comprising: a SIRP-a Dl variant, wherein the SIRP-a Dl variant is a non-naturally occurring high affinity SIRP-a Dl domain, wherein the SIRP-a Dl variant binds to human CD47 with an affinity that is at least 10- fold greater than the affinity of a naturally occurring D l domain; and an Fc domain monomer, wherein the Fc domain monomer is linked to a second polypeptide comprising a second Fc domain monomer to form an Fc domain, wherein the Fc domain has ablated or reduced effector function.
  • the non-naturally occurring high affinity SIRP-a Dl domain comprises an amino acid mutation at residue 80.
  • SIRP-a Dl variant binds CD47 from a first species with a KD less than 250 nM; and wherein the SIRP-a
  • D l variant binds CD47 from a second species with a KD less than 250 nM; and the KD for CD47 from the first species and the KD for CD47 from the second species are within 100 fold of each other; wherein the first species and the second species are selected from the group consisting of: human, rodent, and non-human primate.
  • the SIRP-a Dl variant binds CD47 from at least 3 different species.
  • the non-human primate is cynomolgus monkey. [00183]
  • a polypeptide comprising (a) a SIRP-a
  • the polypeptide is a non-naturally occurring variant of a human SIRP-a.
  • administration of the polypeptide in vivo results in hemoglobin reduction by less than 50% during the first week after administration. In some embodiments, administration of the polypeptide in humans results in hemoglobin reduction by less than 50% during the first week after administration.
  • the polypeptide further comprises at least one Fc variant, wherein the Fc variant comprises an Fc domain monomer selected from (i) a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A; (ii) a human IgG2 Fc region consisting of mutations A330S, P33 IS and N297A; or (iii) a human IgG4 Fc region comprising mutations S228P, E233P, F234V, L235A, delG236, and N297A. .
  • Fc variant comprises an Fc domain monomer selected from (i) a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A; (ii) a human IgG2 Fc region consisting of mutations A330S, P33 IS and N297A; or (iii) a human IgG4 Fc region comprising mutations S
  • the Fc domain monomer is a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A. . In some embodiments, the Fc domain monomer is a human IgG2 Fc region consisting of mutations A330S, P331S and N297A.
  • the SIRP-a constructs of the disclosure include a SIRP-a domain or variant thereof that has its C-terminus joined to the N-terminus of an Fc domain monomer by way of a linker using conventional genetic or chemical means, e.g., chemical conjugation.
  • a linker e.g., a spacer
  • a linker is inserted between the polypeptide and the Fc domain monomer.
  • a polypeptide of the disclosure including a high affinity SIRP-a Dl variant is fused to an Fc domain monomer that is incapable of forming a dimer.
  • a polypeptide of the disclosure is fused to an Fc domain monomer that is capable of forming a dimer, e.g., a heterodimer, with another Fc domain monomer.
  • a polypeptide of the invention is fused to an Fc domain monomer and this fusion protein forms a homodimer.
  • a polypeptide of the disclosure is fused to a first Fc domain monomer and a different protein or peptide (e.g., an antibody variable region) is fused to a second Fc domain monomer.
  • a SIRP-a Dl domain or variant thereof is joined to a first Fc domain monomer and a therapeutic protein (e.g., a cytokine, an interleukin, an antigen, a steroid, an antiinflammatory agent, or an immunomodulatory agent) is joined to a second Fc domain monomer.
  • the first and second Fc domain monomers form a heterodimer.
  • a SIRP-a Dl variant polypeptide (e.g., any of the variants described in Tables 2, 5, and 6) is fused to an Fc polypeptide or Fc variant polypeptide, such as an Fc domain monomer.
  • Fc polypeptide or Fc variant polypeptide such as an Fc domain monomer.
  • polypeptides comprising a SIRP- ⁇ Dl variant polypeptide and a fused Fc variant polypeptide include, but are not limited to,
  • the polypeptide comprises a high affinity SIRP-a Dl domain that has at least 85% sequence identity (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) to any variant provided in Table 8.
  • the polypeptide comprises a high affinity SIRP-a Dl domain that has at least 85% sequence identity (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) to SEQ ID NOs: 98-104, 107-1 13, 1 16-122, or 135-137 in Table 8.
  • the polypeptide comprises (a) a signal-regulatory protein a (SIRP-a) Dl variant, wherein the SIRP-a Dl variant comprises the amino acid sequence,
  • the polypeptide comprises a SIRP-a Dl variant wherein the SIRP-a Dl variant comprises an amino acid sequence according to SEQ ID NO: 47; an Fc variant comprising an Fc domain dimer having two Fc domain monomers, wherein one of the Fc domain monomers in the Fc domain dimer comprises a human IgGl Fc region comprising L234A, L235A, G237A, and N297A mutations.
  • a SIRP-a Dl variant polypeptide (e.g., any of the variants described in Tables 2, 5, and 6) is fused to a first Fc domain monomer either at the N-terminus or at the C-terminus.
  • the first Fc domain monomer is incapable of forming an Fc domain or a dimer.
  • the first Fc domain monomer combines with a second Fc domain monomer to form an Fc domain or a dimer.
  • the first and second Fc domain monomers include amino acid substitutions that promote heterodimerization between the first and second domain monomers.
  • each of the two Fc domain monomers in an Fc domain includes amino acid substitutions that promote the heterodimerization of the two monomers.
  • a SIRP-a construct is formed, for example, from a first subunit including a SIRP-a Dl variant polypeptide fused to a first Fc domain monomer and a second subunit including a second Fc domain monomer (e.g., without a SIRP-a Dl variant polypeptide or any other polypeptide).
  • a construct has a single SIRP-a Dl variant polypeptide linked to an Fc domain (e.g., single arm).
  • a construct has two SIRP-a Dl variant polypeptides linked to an Fc domain (e.g., double arm).
  • a SIRP-a Dl variant having a K D of about 500 nM is particularly useful in a double arm construct.
  • a SIRP-a Dl variant having a K D of about 50 nM is particularly useful in a double arm construct.
  • a SIRP-a Dl variant having a KD of about 5 nM is useful in a double arm construct and a single arm construct.
  • a SIRP-a Dl variant having a K D of about 500 pM is useful in a double arm construct and a single arm construct.
  • a SIRP-a Dl variant having a KD of about 100 pM is useful in a double arm construct and a single arm construct.
  • a SIRP-a Dl variant having a KD of about 50 pM is useful in a double arm construct and a single arm construct. In some embodiments, a SIRP-a Dl variant having a K D of about 10 pM is useful in a double arm construct and a single arm construct.
  • heterodimerization of Fc domain monomers is promoted by introducing different, but compatible, substitutions in the two Fc domain monomers, such as "knob- into-hole” residue pairs and charge residue pairs.
  • the knob and hole interaction favors heterodimer formation, whereas the knob-knob and the hole-hole interaction hinder homodimer formation due to steric clash and deletion of favorable interactions.
  • a hole refers to a void that is created when an original amino acid in a protein is replaced with a different amino acid having a smaller side-chain volume.
  • a knob refers to a bump that is created when an original amino acid in a protein is replaced with a different amino acid having a larger side-chain volume.
  • an amino acid being replaced is in the CH3 antibody constant domain of an Fc domain monomer and involved in the dimerization of two Fc domain monomers.
  • a hole in one CH3 antibody constant domain is created to accommodate a knob in another CH3 antibody constant domain, such that the knob and hole amino acids act to promote or favor the heterodimerization of the two Fc domain monomers.
  • a hole in one CH3 antibody constant domain is created to better accommodate an original amino acid in another CH3 antibody constant domain.
  • a knob in one CH3 antibody constant domain is created to form additional interactions with original amino acids in another CH3 antibody constant domain.
  • a hole is constructed by replacing amino acids having larger side chains such as tyrosine or tryptophan with amino acids having smaller side chains such as alanine, valine, or threonine, for example a Y407V mutation in the CH3 antibody constant domain.
  • a knob is constructed by replacing amino acids having smaller side chains with amino acids having larger side chains, for example a T366W mutation in the CH3 antibody constant domain.
  • one Fc domain monomer includes the knob mutation T366W and the other Fc domain monomer includes hole mutations T366S, L358A, and Y407V.
  • a polypeptide of the disclosure including a high affinity SIRP-a Dl variant is fused to an Fc domain monomer including the knob mutation T366W to limit unwanted knob-knob homodimer formation.
  • knob-into-hole amino acid pairs are included, without limitation, in Table 9 and examples of knob-into-hole Fc variants and SIRP-a - Fc fusions are provided in Table 10.
  • electrostatic steering is also used to control the dimerization of Fc domain monomers.
  • Electrostatic steering refers to the utilization of favorable electrostatic interactions between oppositely charged amino acids in peptides, protein domains, and proteins to control the formation of higher ordered protein molecules.
  • one or more amino acid residues that make up the CH3-CH3 interface are replaced with positively- or negatively-charged amino acid residues such that the interaction becomes electrostatically favorable or unfavorable depending on the specific charged amino acids introduced.
  • a positively-charged amino acid in the interface such as lysine, arginine, or histidine, is replaced with a negatively-charged amino acid such as aspartic acid or glutamic acid.
  • a negatively-charged amino acid in the interface is replaced with a positively-charged amino acid.
  • the charged amino acids are introduced to one of the interacting CH3 antibody constant domains, or both.
  • introducing charged amino acids to the interacting CH3 antibody constant domains of the two Fc domain monomers promotes the selective formation of heterodimers of Fc domain monomers as controlled by the electrostatic steering effects resulting from the interaction between charged amino acids.
  • electrostatic steering amino acid pairs are included, without limitation, in Table 11.
  • a first Fc domain monomer and a second Fc domain monomer each includes one or more of the following amino acid substitutions: T366W, T366S, L368A, Y407V, T366Y, T394W, F405W, Y349T, Y349E, Y349V, L351T, L351H, L351N, L351K, P353S, S354D, D356K, D356R, D356S, E357K, E357R, E357Q, S364A, T366E, L368T, L368Y, L368E, K370E, K370D, K370Q, K392E, K392D, T394N, P395N, P396T, V397T, V397Q, L398T, D399K, D399R, D399N, F405T, F405H,
  • an Fc domain monomer comprises: (a) one of the following amino acid substitutions relative to wild type human IgGl : T366W, T366S, L368A, Y407V, T366Y, T394W, F405W, Y349T, Y349E, Y349V, L351T, L351H, L351N, L351K, P353S, S354D, D356K, D356R, D356S, E357K, E357R, E357Q, S364A, T366E, L368T, L368Y, L368E, K370E, K370D, K370Q, K392E, K392D, T394N, P395N, P396T, V397T, V397Q, L398T, D399K, D399R, D399N, F405T, F405H, F
  • an Fc domain monomer comprises: (a) one of the following amino acid substitutions relative to wild type human IgGl : T366W, T366S, L368A, Y407V, T366Y, T394W, F405W, Y349T, Y349E, Y349V, L351T, L351H, L351N, L351K, P353S, S354D, D356K, D356R, D356S, E357K, E357R, E357Q, S364A, T366E, L368T, L368Y, L368E, K370E, K370D, K370Q, K392E, K392D, T394N, P395N, P396T, V397T, V397Q, L398T, D399K, D399R, D399N, F405T, F405H, F405R,
  • the first and second Fc domain monomers include different amino acid substitutions.
  • the first Fc domain monomer includes T366W.
  • the second Fc domain monomer includes T366S, L368A, and Y407V.
  • the first Fc domain monomer includes D399K.
  • the second Fc domain monomer includes K409D.
  • polypeptides comprising a signal- regulatory protein a (SIRP-a) Dl variant comprising a SIRP-a Dl domain, or a fragment thereof, having an amino acid mutation at residue 80 relative to a wild-type SIRP-a Dl domain; and at least one additional amino acid mutation relative to a wild-type SIRP-a Dl domain at a residue selected from the group consisting of: residue 6, residue 27, residue 31, residue 47, residue 53, residue 54, residue 56, residue 66, and residue 92.
  • SIRP-a signal- regulatory protein a
  • polypeptides comprising an Fc variant, wherein the Fc variant comprises an Fc domain dimer having two Fc domain monomers, wherein each Fc domain monomer independently is selected from (i) a human IgGl Fc region consisting of mutations L234A, L235A, G237A, and N297A; (ii) a human IgG2 Fc region consisting of mutations A330S, P331 S and N297A; or (iii) a human IgG4 Fc region comprising mutations S228P, E233P, F234V, L235A, delG236, and N297A.
  • Fusion to serum albumins can improve the pharmacokinetics of protein
  • Serum albumin is a globular protein that is abundant in blood in mammals. Serum albumin is produced in the liver and can constitute about half of the blood serum proteins. It is monomeric and soluble in the blood. Some of the most crucial functions of serum albumin include transporting hormones, fatty acids, and other proteins in the body, buffering pH, and maintaining osmotic pressure needed for proper distribution of bodily fluids between blood vessels and body tissues. In preferred embodiments, serum albumin is human serum albumin (HSA).
  • HSA human serum albumin
  • an HSA is joined to the C-terminus of the polypeptide of the disclosure to increase the serum half-life of the polypeptide.
  • the N-terminus of an HSA is joined to the C-terminus of the polypeptide of the disclosure.
  • a HSA is joined, either directly or through a linker, to the C-terminus of the polypeptide.
  • an HSA is joined, either directly or through a linker, to the N-terminus of the polypeptide.
  • a human serum albumin comprises the sequence of amino acids (aa) 25-609 of UniProt ID NO: P02768 (SEQ ID NO: 12) as shown in Table 12.
  • the HSA joined to a high affinity SIRP-a Dl variant e.g., any SIRP-a Dl variant described in Tables 2, 5, and 6) includes amino acids 25-609 (SEQ ID NO: 12) of the sequence of UniProt ID NO: P02768.
  • the HSA includes C34S
  • the HSA includes C34S and K573P substitutions, relative to SEQ ID NO: 12.
  • a serum albumin is fused genetically to a polypeptide of the disclosure or joined to the polypeptide through chemical means, e.g., chemical conjugation.
  • a spacer is inserted between the polypeptide and the HSA. Some examples of spacers are described in detail elsewhere herein.
  • a spacer is A or AAAL.
  • the fusion of an HSA in a polypeptide of the disclosure leads to prolonged retention of the polypeptide as well as increases in half-life.
  • Polypeptides comprising a SIRP-a Dl variant polypeptide and a fused HSA include, but are not limited to, SEQ ID NOS: 150-159 provided in Table 13.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Zoology (AREA)
  • Epidemiology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Mycology (AREA)
  • Biomedical Technology (AREA)
  • Oncology (AREA)
  • Endocrinology (AREA)
  • Urology & Nephrology (AREA)
  • Diabetes (AREA)
  • Neurology (AREA)
  • Biotechnology (AREA)
  • Pulmonology (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Hematology (AREA)
  • Rheumatology (AREA)
PCT/US2016/045914 2015-08-07 2016-08-05 Constructs having a sirp-alpha domain or variant thereof Ceased WO2017027422A1 (en)

Priority Applications (33)

Application Number Priority Date Filing Date Title
AU2016304794A AU2016304794B2 (en) 2015-08-07 2016-08-05 Constructs having a SIRP-alpha domain or variant thereof
SI201631278T SI3331902T1 (sl) 2015-08-07 2016-08-05 Konstrukti z domeno SIRP-alfa ali njegove različice
LTEP16835723.4T LT3331902T (lt) 2015-08-07 2016-08-05 Konstruktai, turintys sirp-alfa domeną arba jo variantą
NZ738950A NZ738950A (en) 2015-08-07 2016-08-05 Constructs having a sirp-alpha domain or variant thereof
HK18116580.2A HK1257372B (zh) 2015-08-07 2016-08-05 具有SIRP-α结构域或其变体的构建体
MX2018001428A MX395275B (es) 2015-08-07 2016-08-05 Construcciones con un dominio sirp-alfa o sus variantes
CN202311337928.4A CN117777267A (zh) 2015-08-07 2016-08-05 具有SIRP-α结构域或其变体的构建体
PL16835723T PL3331902T3 (pl) 2015-08-07 2016-08-05 Konstrukty posiadające domenę sirp-alfa lub jej wariant
CN202410176664.7A CN117964732A (zh) 2015-08-07 2016-08-05 具有SIRP-α结构域或其变体的构建体
EP21163641.0A EP3913051A1 (en) 2015-08-07 2016-08-05 Constructs having a sirp-alpha domain or variant thereof
IL302491A IL302491A (en) 2015-08-07 2016-08-05 Structures with a SIRP-alpha site or its variant
HRP20211167TT HRP20211167T1 (hr) 2015-08-07 2016-08-05 Konstrukcije koje imaju sirp-alfa domenu ili njihove varijante
JP2018506281A JP6898303B2 (ja) 2015-08-07 2016-08-05 Ｓｉｒｐ−アルファドメインまたはそのバリアントを有する構築物
IL256989A IL256989B (en) 2015-08-07 2016-08-05 Constructs with a sirp-alpha site or a variant thereof
KR1020187006473A KR102855748B1 (ko) 2015-08-07 2016-08-05 Sirp-알파 도메인 또는 이의 변이체를 갖는 구조체
UAA201802321A UA125638C2 (uk) 2015-08-07 2016-08-05 Конструкції, що мають sirp-альфа домен або його варіант
CN201680057835.8A CN108350048B (zh) 2015-08-07 2016-08-05 具有SIRP-α结构域或其变体的构建体
IL294679A IL294679B2 (en) 2015-08-07 2016-08-05 Structures with a SIRP-alpha site or its variant
DK16835723.4T DK3331902T3 (da) 2015-08-07 2016-08-05 Konstruktioner med et sirp-alpha-domæne eller en variant deraf
SM20210424T SMT202100424T1 (it) 2015-08-07 2016-08-05 Costrutti aventi un dominio sirp-alfa o sua variante
EP16835723.4A EP3331902B1 (en) 2015-08-07 2016-08-05 Constructs having a sirp-alpha domain or variant thereof
BR112018001353-6A BR112018001353B1 (pt) 2015-08-07 2016-08-05 Polipeptídeo compreendendo uma variante d1 da proteína reguladora de sinal a (sirp-a) e uma variante fc, seu método de produção e seus usos, célula hospedeira, e composição farmacêutica
RU2018108103A RU2740672C2 (ru) 2015-08-07 2016-08-05 Конструкции, имеющие sirp-альфа домен или его вариант
KR1020257028619A KR20250133806A (ko) 2015-08-07 2016-08-05 Sirp-알파 도메인 또는 이의 변이체를 갖는 구조체
RS20210912A RS62151B1 (sr) 2015-08-07 2016-08-05 Konstrukti koji sadrže sirp-alfa domen ili njegovu varijantu
ES16835723T ES2881771T3 (es) 2015-08-07 2016-08-05 Constructos que tienen un dominio de SIRP-alfa o variante del mismo
MYPI2018000181A MY185014A (en) 2015-08-07 2016-08-05 Contructs having a sirp-alpha domain or variant thereof
PH1/2018/500269A PH12018500269B1 (en) 2015-08-07 2016-08-05 Constructs having a sirp-alpha domain or variant thereof
CA2993835A CA2993835A1 (en) 2015-08-07 2016-08-05 Constructs having a sirp-alpha domain or variant thereof
ZA2018/00857A ZA201800857B (en) 2015-08-07 2018-02-09 Constructs having a sirp-alpha domain or variant thereof
CONC2018/0002471A CO2018002471A2 (es) 2015-08-07 2018-03-06 Constructos de anticuerpos con un dominio sirp-alfa y sus variantes
CY20211100674T CY1124372T1 (el) 2015-08-07 2021-07-26 Κατασκευασματα που εχουν μια επικρατεια sirp-αλφα ή παραλλαγη αυτης
AU2021218004A AU2021218004B2 (en) 2015-08-07 2021-08-16 Constructs having a SIRP-alpha domain or variant thereof

Applications Claiming Priority (14)

Application Number Priority Date Filing Date Title
US201562202775P 2015-08-07 2015-08-07
US201562202772P 2015-08-07 2015-08-07
US201562202779P 2015-08-07 2015-08-07
US62/202,772 2015-08-07
US62/202,775 2015-08-07
US62/202,779 2015-08-07
US201562265887P 2015-12-10 2015-12-10
US62/265,887 2015-12-10
US201662276801P 2016-01-08 2016-01-08
US201662276796P 2016-01-08 2016-01-08
US62/276,796 2016-01-08
US62/276,801 2016-01-08
US201662346414P 2016-06-06 2016-06-06
US62/346,414 2016-06-06

Publications (1)

Publication Number Publication Date
WO2017027422A1 true WO2017027422A1 (en) 2017-02-16

Family

ID=57983500

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2016/045914 Ceased WO2017027422A1 (en) 2015-08-07 2016-08-05 Constructs having a sirp-alpha domain or variant thereof

Country Status (33)

Country Link
US (5) US10259859B2 (enExample)
EP (2) EP3913051A1 (enExample)
JP (4) JP6898303B2 (enExample)
KR (2) KR102855748B1 (enExample)
CN (3) CN117964732A (enExample)
AU (2) AU2016304794B2 (enExample)
CA (1) CA2993835A1 (enExample)
CL (3) CL2018000340A1 (enExample)
CO (1) CO2018002471A2 (enExample)
CY (1) CY1124372T1 (enExample)
DK (1) DK3331902T3 (enExample)
ES (1) ES2881771T3 (enExample)
GE (2) GEP20227447B (enExample)
HR (1) HRP20211167T1 (enExample)
HU (1) HUE055139T2 (enExample)
IL (3) IL302491A (enExample)
LT (1) LT3331902T (enExample)
MX (2) MX395275B (enExample)
MY (1) MY185014A (enExample)
NZ (1) NZ738950A (enExample)
PE (1) PE20180778A1 (enExample)
PH (1) PH12018500269B1 (enExample)
PL (1) PL3331902T3 (enExample)
PT (1) PT3331902T (enExample)
RS (1) RS62151B1 (enExample)
RU (2) RU2740672C2 (enExample)
SA (1) SA518390881B1 (enExample)
SG (2) SG10201912905VA (enExample)
SI (1) SI3331902T1 (enExample)
SM (1) SMT202100424T1 (enExample)
UA (2) UA125638C2 (enExample)
WO (1) WO2017027422A1 (enExample)
ZA (3) ZA201800857B (enExample)

Cited By (83)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019047885A1 (en) * 2017-09-07 2019-03-14 Dingfu Biotarget Co., Ltd. IMMUNOCONJUGATES COMPRISING ALPHA REGULATORY SIGNAL PROTEIN
US10259859B2 (en) 2015-08-07 2019-04-16 ALX Oncology Inc. Constructs having a SIRP-α domain or variant thereof
CN109971716A (zh) * 2017-12-28 2019-07-05 上海细胞治疗研究院 自分泌cd47抗体的egfr特异性car-t细胞及其用途
WO2019157432A1 (en) 2018-02-12 2019-08-15 Forty Seven, Inc. Anti-cancer regimen using anti-cd47 and anti-cd20 antibodies
WO2019173903A1 (en) * 2018-03-13 2019-09-19 Trillium Therapeutics Inc. Improvements in cd47 blockade therapy by egfr antibody
WO2019234576A1 (en) * 2018-06-03 2019-12-12 Lamkap Bio Beta Ltd. Bispecific antibodies against ceacam5 and cd47
WO2020012486A1 (en) 2018-07-11 2020-01-16 Kahr Medical Ltd. SIRPalpha-4-1BBL VARIANT FUSION PROTEIN AND METHODS OF USE THEREOF
WO2020047326A2 (en) 2018-08-31 2020-03-05 ALX Oncology Inc. Decoy polypeptides
JP2020508329A (ja) * 2017-02-27 2020-03-19 シャタック ラボ，インコーポレイテッド 細胞外ドメインベースのキメラタンパク質の製造方法及び使用方法
CN110958888A (zh) * 2017-03-28 2020-04-03 延龄草治疗公司 Cd47阻断疗法
WO2020086758A1 (en) * 2018-10-23 2020-04-30 Dragonfly Therapeutics, Inc. Heterodimeric fc-fused proteins
US10696722B2 (en) 2016-08-10 2020-06-30 Ajou University Industry-Academic Cooperation Foundation Heterodimeric Fc-fused cytokine and pharmaceutical composition comprising the same
WO2020200941A1 (en) 2019-03-29 2020-10-08 F. Hoffmann-La Roche Ag Spr-based binding assay for the functional analysis of multivalent molecules
CN111836647A (zh) * 2018-03-13 2020-10-27 延龄草治疗公司 Cd47阻断疗法和cd38抗体的组合
WO2020243338A1 (en) * 2019-05-31 2020-12-03 ALX Oncology Inc. Methods of treating cancer with sirp alpha fc fusion in combination with an immune checkpoint inhibitor
WO2020247820A1 (en) 2019-06-07 2020-12-10 ALX Oncology Inc. Methods and reagents for reducing the interference of drugs that bind cd47 in serological assays
WO2021011544A1 (en) 2019-07-16 2021-01-21 Gilead Sciences, Inc. Hiv vaccines and methods of making and using
JP2021505195A (ja) * 2017-12-04 2021-02-18 北京韓美薬品有限公司Ｂｅｉｊｉｎｇ Ｈａｎｍｉ Ｐｈａｒｍ． Ｃｏ．， Ｌｔｄ． 天然抗体様構造を有し、ヘテロ二量体形態の抗ｐｄ−ｌ１／抗ｃｄ４７二重特異性抗体、ならびにその製造方法
US10927173B2 (en) 2016-01-11 2021-02-23 Forty Seven, Inc. Humanized, mouse or chimeric anti-CD47 monoclonal antibodies
WO2021076908A1 (en) 2019-10-18 2021-04-22 Forty Seven, Inc. Combination therapies for treating myelodysplastic syndromes and acute myeloid leukemia
WO2021087064A1 (en) 2019-10-31 2021-05-06 Forty Seven, Inc. Anti-cd47 and anti-cd20 based treatment of blood cancer
JP2021511817A (ja) * 2017-11-20 2021-05-13 泰州▲邁▼博太科▲薬▼▲業▼有限公司Ｔａｉｚｈｏｕ Ｍａｂｔｅｃｈ Ｐｈａｒｍａｃｅｕｔｉｃａｌ Ｃｏ．， Ｌｔｄ Ｃｄ４７とｐｄ−ｌ１を標的にする二重機能の融合タンパク質
US20210154269A1 (en) * 2019-11-27 2021-05-27 ALX Oncology Inc. Combination therapies for treating cancer
WO2021130638A1 (en) 2019-12-24 2021-07-01 Carna Biosciences, Inc. Diacylglycerol kinase modulating compounds
EP3341015B1 (en) 2015-08-26 2021-07-28 The Board of Trustees of the Leland Stanford Junior University Enhanced depletion of targeted cells with cd47 blockade and an immune costimulatory agonist
EP3623388A4 (en) * 2017-05-08 2021-08-04 Shanghai JMT-Bio Technology Co., Ltd. BISPECIFIC RECOMBINANT PROTEIN AND ITS USE
WO2021163064A2 (en) 2020-02-14 2021-08-19 Jounce Therapeutics, Inc. Antibodies and fusion proteins that bind to ccr8 and uses thereof
JP2021520830A (ja) * 2018-04-17 2021-08-26 杭州尚健生物技術有限公司 Ｃｄ４７タンパク質に結合する融合タンパク質およびその使用
US11130796B2 (en) 2017-01-05 2021-09-28 Kahr Medical Ltd. SIRPalpha-41BBL fusion protein and methods of use thereof
WO2021247430A1 (en) 2020-06-01 2021-12-09 ALX Oncology Inc. Combination therapies comprising a hypomethylation agent for treating cancer
WO2022010806A1 (en) 2020-07-06 2022-01-13 ALX Oncology Inc. Methods for reducing the interference of drugs that bind therapeutic targets expressed on blood cells in serological assays
US11299530B2 (en) 2017-01-05 2022-04-12 Kahr Medical Ltd. SIRP alpha-CD70 fusion protein and methods of use thereof
WO2022120286A1 (en) 2020-12-06 2022-06-09 ALX Oncology Inc. Multimers for reducing the interference of drugs that bind cd47 in serological assays
WO2022190058A1 (en) 2021-03-12 2022-09-15 Dcprime B.V. Methods of vaccination and use of cd47 blockade
WO2022221304A1 (en) 2021-04-14 2022-10-20 Gilead Sciences, Inc. CO-INHIBITION OF CD47/SIRPα BINDING AND NEDD8-ACTIVATING ENZYME E1 REGULATORY SUBUNIT FOR THE TREATMENT OF CANCER
WO2022241157A1 (en) 2021-05-13 2022-11-17 ALX Oncology Inc. Combination therapies for treating cancer
WO2022271684A1 (en) 2021-06-23 2022-12-29 Gilead Sciences, Inc. Diacylglyercol kinase modulating compounds
WO2022271650A1 (en) 2021-06-23 2022-12-29 Gilead Sciences, Inc. Diacylglyercol kinase modulating compounds
WO2022271659A1 (en) 2021-06-23 2022-12-29 Gilead Sciences, Inc. Diacylglyercol kinase modulating compounds
WO2022271677A1 (en) 2021-06-23 2022-12-29 Gilead Sciences, Inc. Diacylglyercol kinase modulating compounds
RU2787521C2 (ru) * 2018-04-17 2023-01-10 Ханчжоу Сумген Байотек Ко., Лтд. Связывание слитого белка с белком cd47 и его применение
US11566060B2 (en) 2017-01-05 2023-01-31 Kahr Medical Ltd. PD1-CD70 fusion protein and methods of use thereof
WO2023077030A1 (en) 2021-10-29 2023-05-04 Gilead Sciences, Inc. Cd73 compounds
WO2023076983A1 (en) 2021-10-28 2023-05-04 Gilead Sciences, Inc. Pyridizin-3(2h)-one derivatives
WO2023122615A1 (en) 2021-12-22 2023-06-29 Gilead Sciences, Inc. Ikaros zinc finger family degraders and uses thereof
WO2023122581A2 (en) 2021-12-22 2023-06-29 Gilead Sciences, Inc. Ikaros zinc finger family degraders and uses thereof
US11702458B2 (en) 2017-01-05 2023-07-18 Kahr Medical Ltd. PD1-41BBL fusion protein and methods of use thereof
WO2023147418A1 (en) 2022-01-28 2023-08-03 Gilead Sciences, Inc. Parp7 inhibitors
WO2023154578A1 (en) 2022-02-14 2023-08-17 Sana Biotechnology, Inc. Methods of treating patients exhibiting a prior failed therapy with hypoimmunogenic cells
EP3980051A4 (en) * 2019-06-05 2023-09-13 Asher Biotherapeutics, Inc. FUSIONS OF MUTANT INTERLEUKIN-2 POLYPEPTIDES WITH ANTIGEN-BINDING MOLECULES TO MODULATE THE FUNCTION OF IMMUNE CELLS
EP4245756A1 (en) 2022-03-17 2023-09-20 Gilead Sciences, Inc. Ikaros zinc finger family degraders and uses thereof
WO2023183890A1 (en) 2022-03-24 2023-09-28 Bitterroot Bio, Inc. Multivalent sirp-alpha fusion polypeptides
WO2023183817A1 (en) 2022-03-24 2023-09-28 Gilead Sciences, Inc. Combination therapy for treating trop-2 expressing cancers
WO2023183892A1 (en) 2022-03-24 2023-09-28 Bitterroot Bio, Inc. Sirp-alpha fusion polypeptides with modified fc domains
WO2023183313A1 (en) 2022-03-22 2023-09-28 Sana Biotechnology, Inc. Engineering cells with a transgene in b2m or ciita locus and associated compositions and methods
WO2023196784A1 (en) 2022-04-05 2023-10-12 Gilead Sciences, Inc. Combinations of antibody therapies for treating colorectal cancer
WO2023205719A1 (en) 2022-04-21 2023-10-26 Gilead Sciences, Inc. Kras g12d modulating compounds
WO2023218378A1 (en) 2022-05-11 2023-11-16 Daiichi Sankyo Company, Limited Combination of an antibody specific for a tumor antigen and a cd47 inhibitor
WO2023235754A1 (en) 2022-06-01 2023-12-07 ALX Oncology Inc. Combination therapies for treating urothelial carcinoma
WO2024006929A1 (en) 2022-07-01 2024-01-04 Gilead Sciences, Inc. Cd73 compounds
WO2024015741A1 (en) 2022-07-12 2024-01-18 Gilead Sciences, Inc. Hiv immunogenic polypeptides and vaccines and uses thereof
WO2024064668A1 (en) 2022-09-21 2024-03-28 Gilead Sciences, Inc. FOCAL IONIZING RADIATION AND CD47/SIRPα DISRUPTION ANTICANCER COMBINATION THERAPY
EP4155318A4 (en) * 2020-06-30 2024-06-05 Harbour Biomed US, Inc. BISPECIFIC ANTIBODY AND USE THEREOF
WO2024137852A1 (en) 2022-12-22 2024-06-27 Gilead Sciences, Inc. Prmt5 inhibitors and uses thereof
US12024559B2 (en) 2020-10-23 2024-07-02 Asher Biotherapeutics, Inc. Fusions with CD8 antigen binding molecules for modulating immune cell function
EP4190803A4 (en) * 2020-07-30 2024-08-21 Sunshine Guojian Pharmaceutical (Shanghai) Co., Ltd. SIRPa-FC FUSION PROTEIN
US12091681B2 (en) 2020-03-27 2024-09-17 Mendus B.V. Ex vivo use of modified cells of leukemic origin for enhancing the efficacy of adoptive cell therapy
WO2024215754A1 (en) 2023-04-11 2024-10-17 Gilead Sciences, Inc. Kras modulating compounds
US12122827B2 (en) 2021-05-19 2024-10-22 Asher Biotherapeutics, Inc. IL-21 polypeptides and targeted constructs
WO2024220917A1 (en) 2023-04-21 2024-10-24 Gilead Sciences, Inc. Prmt5 inhibitors and uses thereof
US20240400691A1 (en) * 2023-05-31 2024-12-05 Fbd Biologics Limited Cd47/pd-l1-targeting protein complex and methods of use thereof
WO2025006720A1 (en) 2023-06-30 2025-01-02 Gilead Sciences, Inc. Kras modulating compounds
WO2025024663A1 (en) 2023-07-26 2025-01-30 Gilead Sciences, Inc. Parp7 inhibitors
WO2025024811A1 (en) 2023-07-26 2025-01-30 Gilead Sciences, Inc. Parp7 inhibitors
WO2025054530A1 (en) 2023-09-08 2025-03-13 Gilead Sciences, Inc. Pyrimidine-containing polycyclic derivatives as kras g12d modulating compounds
WO2025054347A1 (en) 2023-09-08 2025-03-13 Gilead Sciences, Inc. Kras g12d modulating compounds
US12286466B2 (en) 2018-07-11 2025-04-29 Kahr Medical Ltd. PD1-4-1BBL variant fusion protein and methods of use thereof
WO2025096589A1 (en) 2023-11-03 2025-05-08 Gilead Sciences, Inc. Prmt5 inhibitors and uses thereof
US12304967B2 (en) 2018-11-14 2025-05-20 Hangzhou Sumgen Biotech Co., Ltd. Fusion protein and use thereof
WO2025137640A1 (en) 2023-12-22 2025-06-26 Gilead Sciences, Inc. Azaspiro wrn inhibitors
US12364758B2 (en) 2020-06-30 2025-07-22 Mendus B.V. Use of leukemia-derived cells in ovarian cancer vaccines
EP4319796A4 (en) * 2021-12-21 2025-07-23 Fbd Biologics Ltd MODIFIED SIRPalpha VARIANTS AND METHODS OF USE THEREOF
WO2025245003A1 (en) 2024-05-21 2025-11-27 Gilead Sciences, Inc. Prmt5 inhibitors and uses thereof

Families Citing this family (48)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10053513B2 (en) * 2009-11-30 2018-08-21 Janssen Biotech, Inc. Antibody Fc mutants with ablated effector functions
US12466897B2 (en) 2011-10-10 2025-11-11 Xencor, Inc. Heterodimeric human IgG1 polypeptides with isoelectric point modifications
US10858417B2 (en) 2013-03-15 2020-12-08 Xencor, Inc. Heterodimeric proteins
CN116333153A (zh) 2014-11-26 2023-06-27 森科股份有限公司 结合cd3和肿瘤抗原的异二聚体抗体
US20170151281A1 (en) 2015-02-19 2017-06-01 Batu Biologics, Inc. Chimeric antigen receptor dendritic cell (car-dc) for treatment of cancer
CN116063566A (zh) 2015-10-01 2023-05-05 热生物制品有限公司 作为异源嵌合蛋白邻接i型和ii型胞外结构域的组合物和方法
US11560433B2 (en) 2016-05-27 2023-01-24 Albert Einstein College Of Medicine Methods of treatment by targeting VCAM1 and MAEA
EP3577133A1 (en) 2017-02-06 2019-12-11 Orionis Biosciences NV Targeted chimeric proteins and uses thereof
CN110381974A (zh) 2017-02-27 2019-10-25 沙塔克实验室有限公司 基于csf1r的嵌合蛋白
AU2018281337B2 (en) * 2017-06-06 2022-08-25 Relinia, Inc. Single-chain TNF receptor 2 agonist fusion proteins
AU2019269641A1 (en) * 2018-05-17 2020-12-03 Immunome, Inc. CH3 domain epitope tags
US12084497B2 (en) 2018-08-08 2024-09-10 Orionis Biosciences, Inc. SIRP1α targeted chimeric proteins and uses thereof
US10780121B2 (en) 2018-08-29 2020-09-22 Shattuck Labs, Inc. FLT3L-based chimeric proteins
US11591390B2 (en) 2018-09-27 2023-02-28 Celgene Corporation SIRP-α binding proteins and methods of use thereof
BR112021005585A2 (pt) 2018-09-27 2021-06-29 Celgene Corporation proteínas de ligação a sirpa e métodos de uso das mesmas
US12331320B2 (en) 2018-10-10 2025-06-17 The Research Foundation For The State University Of New York Genome edited cancer cell vaccines
CN109306017B (zh) * 2018-10-12 2021-02-09 倍而达药业（苏州）有限公司 一种基于SIRP-αD1突变体制备的重组蛋白及应用
EP3876977A1 (en) 2018-11-06 2021-09-15 The Regents Of The University Of California Chimeric antigen receptors for phagocytosis
CN109517054B (zh) * 2018-12-04 2022-04-08 江苏东抗生物医药科技有限公司 SIRPα变体或其融合蛋白及其应用
MX2021010531A (es) 2019-03-06 2021-10-01 Jiangsu Hengrui Medicine Co Proteina de fusion bifuncional y uso farmaceutico de la misma.
CN111763261B (zh) * 2019-04-02 2022-08-09 杭州尚健生物技术有限公司 一种融合蛋白及其用途
US11013764B2 (en) 2019-04-30 2021-05-25 Myeloid Therapeutics, Inc. Engineered phagocytic receptor compositions and methods of use thereof
CN112125975B (zh) * 2019-06-25 2024-03-01 上海翰森生物医药科技有限公司 Pd-l1和cd47双特异性融合蛋白及其医药用途
KR102228271B1 (ko) * 2019-07-16 2021-03-17 한국과학기술연구원 항암활성을 갖는 면역조절 단백질-siRNA 복합체
KR20220097875A (ko) 2019-09-03 2022-07-08 마이얼로이드 테라퓨틱스, 인크. 게놈 통합을 위한 방법 및 조성물
US10980836B1 (en) 2019-12-11 2021-04-20 Myeloid Therapeutics, Inc. Therapeutic cell compositions and methods of manufacturing and use thereof
CN115315273A (zh) 2020-01-14 2022-11-08 辛德凯因股份有限公司 Il-2直向同源物及其使用方法
CN111253482B (zh) * 2020-02-18 2021-11-30 中国人民解放军军事科学院军事医学研究院 SIRPa变体、融合蛋白、及其应用
WO2021231976A1 (en) 2020-05-14 2021-11-18 Xencor, Inc. Heterodimeric antibodies that bind prostate specific membrane antigen (psma) and cd3
CN113912736A (zh) * 2020-07-09 2022-01-11 上海交通大学 靶向cd47与pd-l1双效融合蛋白及其应用
CN111808183B (zh) * 2020-07-25 2022-07-08 北京吉尔麦迪生物医药科技有限公司 一种靶向CD47的高亲和力SIRPα突变体及其融合蛋白
JP7538334B2 (ja) * 2020-08-14 2024-08-21 コリア・インスティテュート・オブ・サイエンス・アンド・テクノロジー 抗癌活性を有する免疫調節タンパク質－ｓｉＲＮＡ複合体
KR102579284B1 (ko) * 2020-09-22 2023-09-18 비피진 주식회사 신규의 cd47 바인더와 폴리뉴클레오티드를 포함하는 암 치료를 위한 리포좀 복합체
KR102557016B1 (ko) * 2020-09-22 2023-07-20 비피진 주식회사 암 치료를 위한 cd47 바인더 및 리포좀 복합체
MX2023005201A (es) 2020-11-04 2023-06-28 Myeloid Therapeutics Inc Composiciones de proteinas de fusion quimerica modificadas por ingenieria y metodos de uso de las mismas.
CN112979782B (zh) * 2021-03-08 2023-07-18 深圳市乐土生物医药有限公司 一种多肽及其用途
JP2024511319A (ja) * 2021-03-09 2024-03-13 ゼンコア インコーポレイテッド Ｃｄ３及びｃｌｄｎ６に結合するヘテロ二量体抗体
EP4337268A4 (en) 2021-05-11 2025-06-04 Myeloid Therapeutics, Inc. Methods and compositions for genomic integration
KR20230016152A (ko) * 2021-07-19 2023-02-01 주식회사유한양행 Sirp-alpha 변이체 및 이의 용도
CN113896802A (zh) * 2021-10-09 2022-01-07 宜明昂科生物医药技术（上海）有限公司 靶向cd47和cd38的重组融合蛋白及其制备和用途
WO2023072279A1 (zh) * 2021-11-01 2023-05-04 江苏先声药业有限公司 SIRPa突变体及其应用
CN116178561B (zh) * 2021-11-26 2025-07-18 杭州尚健生物技术有限公司 包含SIRPα突变体的融合蛋白
EP4434999A1 (en) * 2021-11-19 2024-09-25 Hangzhou Sumgen Biotech Co., Ltd. Sirp? variant and use thereof
US20240043846A1 (en) 2021-11-19 2024-02-08 Kist (Korea Institute Of Science And Technology) Therapeutic Compounds for Red Blood Cell-Mediated Delivery of an Active Pharmaceutical Ingredient to a Target Cell
KR102710779B1 (ko) * 2022-01-04 2024-09-26 주식회사 보령바이오파마 5'-뉴클레오티다아제 변형 단백질을 암호화하는 암 치료를 위한 폴리뉴클레오티드
CN116041470B (zh) * 2022-08-04 2024-10-22 四川大学华西医院 具有高亲和力结合肿瘤细胞cd47的细胞膜材料及其制备方法和应用
WO2024053788A1 (ko) * 2022-09-07 2024-03-14 주식회사 시프트바이오 SIRPΑα 변이체에 특이적으로 결합하는 항체
WO2025072726A1 (en) 2023-09-29 2025-04-03 Trex Bio, Inc. Tnf-alpha variant fusion molecules

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010070047A1 (en) 2008-12-19 2010-06-24 Novartis Ag Soluble polypeptides for use in treating autoimmune and inflammatory disorders
US20100239579A1 (en) * 2006-05-15 2010-09-23 Viral Logic Systems Technology Corp. CD47 Related Compositions and Methods for Treating Immunological Diseases and Disorders
WO2012142515A2 (en) * 2011-04-13 2012-10-18 Bristol-Myers Squibb Company Fc fusion proteins comprising novel linkers or arrangements
WO2013109752A1 (en) 2012-01-17 2013-07-25 The Board Of Trustees Of The Leland Stanford Junior University High affinity sirp-alpha reagents
WO2016023040A1 (en) * 2014-08-08 2016-02-11 Alexo Therapeutics International Sirp-alpha variant constructs and uses thereof

Family Cites Families (102)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3773919A (en) 1969-10-23 1973-11-20 Du Pont Polylactide-drug mixtures
US5116964A (en) 1989-02-23 1992-05-26 Genentech, Inc. Hybrid immunoglobulins
US5697901A (en) 1989-12-14 1997-12-16 Elof Eriksson Gene delivery by microneedle injection
FI101678B1 (fi) 1990-12-31 1998-08-14 Akzo Nv Happolabiileja kytkentämolekyylejä
CA2110899C (en) 1991-06-21 2006-08-08 Jacob G. Michael Orally administrable therapeutic proteins and method of making
US6613332B1 (en) 1991-06-21 2003-09-02 The University Of Cincinnati Oral administration of therapeutic proteins
IL99120A0 (en) 1991-08-07 1992-07-15 Yeda Res & Dev Multimers of the soluble forms of tnf receptors,their preparation and pharmaceutical compositions containing them
WO1993006217A1 (en) 1991-09-19 1993-04-01 Genentech, Inc. EXPRESSION IN E. COLI OF ANTIBODY FRAGMENTS HAVING AT LEAST A CYSTEINE PRESENT AS A FREE THIOL, USE FOR THE PRODUCTION OF BIFUNCTIONAL F(ab')2 ANTIBODIES
US5505931A (en) 1993-03-04 1996-04-09 The Dow Chemical Company Acid cleavable compounds, their preparation and use as bifunctional acid-labile crosslinking agents
US5731168A (en) 1995-03-01 1998-03-24 Genentech, Inc. Method for making heteromultimeric polypeptides
JPH11510050A (ja) 1995-07-25 1999-09-07 イントロヘーネ ベスローテン フェンノートシャップ 標的遺伝子送達のための方法および手段
CA2246332C (en) 1996-02-15 2009-04-14 Biosense, Inc. Catheter based surgery
EP0920339A2 (en) 1996-07-09 1999-06-09 The Johns Hopkins University Gene delivery system
US6541615B1 (en) 1996-11-15 2003-04-01 Max-Planck-Gellschaft Zur Foderung Der Wissenschaften E.V. SIRP proteins and uses thereof
CA2226962A1 (en) 1998-02-16 1999-08-16 Marie Sarfati Use of binding agents to cd47 and its ligands in the treatment or the prophylaxis of various inflammatory, autoimmune and allergic diseases and in the treatment of graft rejection
JP2003502062A (ja) 1999-06-11 2003-01-21 ヒューマン ジノーム サイエンシーズ， インコーポレイテッド ４９個のヒト分泌タンパク質
US6613026B1 (en) 1999-12-08 2003-09-02 Scimed Life Systems, Inc. Lateral needle-less injection apparatus and method
GB9930706D0 (en) 1999-12-24 2000-02-16 Medical Res Council Composition for inhibiting macrophage activity
US7662367B2 (en) 2000-03-02 2010-02-16 Xencor, Inc. Pharmaceutical compositions for the treatment of TNF-α related disorders
TWI334439B (en) 2001-08-01 2010-12-11 Centocor Inc Anti-tnf antibodies, compositions, methods and uses
GB0124145D0 (en) 2001-10-08 2001-11-28 Bayer Ag Genes and proteins for prevention,prediction,prognosis and therapy of cardiovascular disease
US20040018522A1 (en) 2002-05-09 2004-01-29 Brigham And Women's Hospital, Inc. Identification of dysregulated genes in patients with multiple sclerosis
AU2003259285A1 (en) 2002-07-29 2004-02-16 Hmgene, Inc. Methods of identifying adipocyte specific genes, the genes identified, and their uses
JP2006515165A (ja) 2002-09-16 2006-05-25 ジェネンテック・インコーポレーテッド 免疫関連疾患の治療のための新規組成物と方法
EP3299393A1 (en) 2002-11-08 2018-03-28 Ablynx N.V. Single domain antibodies directed against tumour necrosis factor-alpha and uses therefor
US8613922B2 (en) 2003-04-24 2013-12-24 The University Of North Carolina At Chapel Hill Methods for inhibiting diabetic retinopathy with an antibody against integrin associated protein (IAP)
US20040213792A1 (en) 2003-04-24 2004-10-28 Clemmons David R. Method for inhibiting cellular activation by insulin-like growth factor-1
US20100215640A1 (en) 2003-04-24 2010-08-26 The University Of North Carolina At Chapel Hill Method for inhibiting cellular activation by insulin-like growth factor-1
US7691970B2 (en) 2003-08-25 2010-04-06 Pieris Ag Muteins of a bilin-binding protein with affinity for a given target
US7892558B2 (en) 2004-02-27 2011-02-22 Vaxconsulting Isolated TNF-alpha peptide and pharmaceutical composition thereof
US8518869B2 (en) 2004-03-29 2013-08-27 Austria Wirtschaftsservice Gesellschaft M.B.H. Pharmaceutical combined preparation containing a therapeutic protein
WO2005108415A2 (en) 2004-04-30 2005-11-17 Biogen Idec Ma Inc. Membrane associated molecules
US7514229B2 (en) 2005-09-29 2009-04-07 The Board Of Trustees Of The Leland Stanford Junior University Methods for diagnosing and evaluating treatment of blood disorders
AR060017A1 (es) * 2006-01-13 2008-05-21 Novartis Ag Composiciones y metodos de uso para anticuerpos de dickkopf -1
US20080260738A1 (en) 2007-04-18 2008-10-23 Moore Margaret D Single chain fc, methods of making and methods of treatment
RU2009140134A (ru) * 2007-04-23 2011-05-27 Вайет (Us) Способы и композиции для лечения и мониторинга лечения связанных с ил-13 нарушений
EP3492488A1 (en) 2007-08-22 2019-06-05 The Regents of The University of California Activatable binding polypeptides and methods of identification and use thereof
CN101970478A (zh) 2007-10-11 2011-02-09 大学健康网络 调节SIRPα-CD47相互作用以增加造血干细胞植入以及用于此的化合物
JO3076B1 (ar) 2007-10-17 2017-03-15 Janssen Alzheimer Immunotherap نظم العلاج المناعي المعتمد على حالة apoe
PT2235064E (pt) 2008-01-07 2016-03-01 Amgen Inc Método de preparação de moléculas heterodiméricas de fc de anticorpos utilizando efeitos de indução eletrostática
PT3056515T (pt) 2008-01-15 2019-07-19 Univ Leland Stanford Junior Métodos para manipulação da fagocitose mediada por cd47
EP2111869A1 (en) 2008-04-23 2009-10-28 Stichting Sanquin Bloedvoorziening Compositions and methods to enhance the immune system
WO2010081173A2 (en) 2009-01-12 2010-07-15 Cytomx Therapeutics, Llc Modified antibody compositions, methods of making and using thereof
AU2010215761B2 (en) 2009-02-23 2017-04-06 Cytomx Therapeutics, Inc Proproteins and methods of use thereof
PL2429574T3 (pl) 2009-05-15 2015-12-31 Univ Health Network Kompozycje i sposoby leczenia nowotworów hematologicznych celujące w oddziaływanie SIRP-CD47
MX2012000895A (es) 2009-07-20 2012-06-01 Univ Nat Cheng Kung Polipeptidos selectivos para integrina alfavbeta3 conjugada con una variante de albumina de suero humana (hsa) y usos farmaceuticos de los mismos.
KR101632312B1 (ko) 2009-11-03 2016-06-21 삼성전자주식회사 항체 불변 영역에 특이적으로 결합하는 융합 단백질, 그의 제조 방법 및 이를 이용한 항체 분리 방법
SI2506871T1 (sl) * 2009-11-30 2016-12-30 Janssen Biotech, Inc. Mutanti Fc protitelesa z odstranjenimi efektorskimi funkcijami
AU2010334974A1 (en) 2009-12-22 2012-07-12 Novartis Ag Tetravalent CD47-antibody constant region fusion protein for use in therapy
HRP20170254T1 (hr) 2010-05-14 2017-04-21 The Board of Trustees of the Leland Stanford Junior University Office of the General Counsel Humanizirana i kimerna monoklonska protutijela usmjerena na cd47
EP2624868B1 (en) 2010-10-08 2018-09-26 City of Hope A monoclonal antibody framework binding interface for meditopes, meditope delivery systems and methods for their use
US8609621B2 (en) 2010-11-15 2013-12-17 E I Du Pont De Nemours And Company Acid-cleavable linkers exhibiting altered rates of acid hydrolysis
US9566347B2 (en) 2011-02-07 2017-02-14 The Trustees Of The University Of Pennsylvania Peptides and methods using same
KR20160044598A (ko) 2011-03-29 2016-04-25 로슈 글리카트 아게 항체 Fc 변이체
KR20140059168A (ko) 2011-04-21 2014-05-15 더 리젠츠 오브 더 유니버시티 오브 콜로라도, 어 바디 코포레이트 시신경 척수염 치료용 조성물 및 치료 방법
SG194510A1 (en) * 2011-04-22 2013-12-30 Emergent Product Dev Seattle Prostate-specific membrane antigen binding proteins and related compositionsand methods
WO2012172521A1 (en) 2011-06-16 2012-12-20 Novartis Ag Soluble proteins for use as therapeutics
US20140242095A1 (en) 2011-10-19 2014-08-28 University Health Network Antibodies and antibody fragments targeting sirp-alpha and their use in treating hematologic cancers
WO2013063076A1 (en) 2011-10-25 2013-05-02 Indiana University Research & Technology Corporation Compositions for and methods of modulating complications, risks and issues with xenotransplantation
US9428553B2 (en) 2012-02-10 2016-08-30 City Of Hope Meditopes and meditope-binding antibodies and uses thereof
CA2876904C (en) 2012-06-22 2019-12-03 Cytomx Therapeutics, Inc. Anti-jagged 1/jagged 2 cross-reactive antibodies, activatable anti-jagged antibodies and methods of use thereof
US9856314B2 (en) 2012-06-22 2018-01-02 Cytomx Therapeutics, Inc. Activatable antibodies having non-binding steric moieties and methods of using the same
GB201216649D0 (en) 2012-09-18 2012-10-31 Univ Birmingham Agents and methods
JP6335189B2 (ja) 2012-12-17 2018-05-30 トリリアム・セラピューティクス・インコーポレイテッドＴｒｉｌｌｉｕｍ Ｔｈｅｒａｐｅｕｔｉｃｓ Ｉｎｃ． ＳＩＲＰアルファ−Ｆｃ融合体でのＣＤ４７＋疾患細胞の治療
US9873747B2 (en) 2013-01-31 2018-01-23 Thomas Jefferson University Fusion proteins that facilitate cancer cell destruction
PL4119153T3 (pl) 2013-02-05 2024-09-02 The Board Of Trustees Of The Leland Stanford Junior University Terapie ukierunkowane na cd47 w leczeniu choroby zakaźnej
KR102276974B1 (ko) 2013-02-06 2021-07-13 인히브릭스, 인크. 비-혈소판 및 비-적혈구 세포 격감성 cd47 항체 및 이를 이용하는 방법
EP2968536B1 (en) 2013-03-13 2023-06-28 The United States of America, as represented by The Secretary, Department of Health and Human Services Methods for modulating chemotherapeutic cytotoxicity
JP6426693B2 (ja) 2013-03-15 2018-11-21 ザ ボード オブ トラスティーズ オブ ザ レランド スタンフォード ジュニア ユニバーシティー 抗ｃｄ４７薬の処理上有効量を達成するための方法
WO2014179132A1 (en) 2013-04-29 2014-11-06 The Board Of Trustees Of The Leland Stanford Junior University Use of anti-cd47 agents to enhance immunization
US9945870B2 (en) 2013-05-17 2018-04-17 The Board Of Trustees Of The Leland Stanford Junior University Methods for determining responsiveness to an anti-CD47 agent
KR20250152665A (ko) * 2013-07-31 2025-10-23 암젠 인크 Ｆc-함유 폴리펩타이드의 안정화
US10329354B2 (en) 2013-09-18 2019-06-25 The Board Of Trustees Of The Leland Stanford Junior University Modulation of efferocytosis pathways for treatment of atherosclerotic disease
SMT202500280T1 (it) 2013-09-23 2025-09-12 Regeneron Pharma Animali non umani aventi un gene umanizzato della proteina regolatrice del segnale
IL291329B1 (en) 2013-09-25 2025-09-01 Cytomx Therapeutics Inc Polypeptides that can be determined in matrix metalloproteinases and uses thereof
CA2927543C (en) * 2013-10-15 2021-07-20 The California Institute For Biomedical Research Peptidic chimeric antigen receptor t cell switches and uses thereof
CN106459153B (zh) 2014-01-31 2021-12-21 西托姆克斯治疗公司 蛋白裂解酶和u型纤溶酶原激活物的底物和其它可裂解部分及其使用方法
ES3040431T3 (en) 2014-03-12 2025-10-31 Yeda Res & Dev Reducing systemic regulatory t cell levels or activity for treatment of disease and injury of the cns
US10519237B2 (en) 2014-03-12 2019-12-31 Yeda Research And Development Co. Ltd Reducing systemic regulatory T cell levels or activity for treatment of disease and injury of the CNS
US9394365B1 (en) 2014-03-12 2016-07-19 Yeda Research And Development Co., Ltd Reducing systemic regulatory T cell levels or activity for treatment of alzheimer's disease
EP3177640B1 (en) 2014-08-08 2020-05-06 The Board of Trustees of the Leland Stanford Junior University High affinity pd-1 agents and methods of use
WO2016023001A1 (en) 2014-08-08 2016-02-11 The Board Of Trustees Of The Leland Stanford Junior University Multispecific high affinity pd-1 agents and methods of use
US10087257B2 (en) 2014-08-08 2018-10-02 The Board Of Trustees Of The Leland Stanford Junior University SIRP alpha-antibody fusion proteins
WO2016024021A1 (en) 2014-08-15 2016-02-18 Merck Patent Gmbh Sirp-alpha immunoglobulin fusion proteins
ES2945588T3 (es) 2014-08-26 2023-07-04 Univ Leland Stanford Junior Injerto de células madre con una combinación de un agente dirigido a las células madre y modulación de señalización inmunoreguladora
WO2016044021A1 (en) 2014-09-15 2016-03-24 The Board Of Trustees Of The Leland Stanford Junior University Targeting aneurysm disease by modulating phagocytosis pathways
CA2964173A1 (en) 2014-10-10 2016-04-14 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Methods to eliminate cancer stem cells by targeting cd47
EP3209769B1 (en) 2014-10-24 2020-08-05 The Board of Trustees of the Leland Stanford Junior University Compositions and methods for inducing phagocytosis of mhc class i positive cells and countering anti-cd47/sirpa resistance
EP3012271A1 (en) 2014-10-24 2016-04-27 Effimune Method and compositions for inducing differentiation of myeloid derived suppressor cell to treat cancer and infectious diseases
AU2015350190B2 (en) 2014-11-18 2021-08-05 Janssen Pharmaceutica Nv CD47 antibodies, methods, and uses
EP4218812A1 (en) 2015-02-27 2023-08-02 The Board of Trustees of the Leland Stanford Junior University Combination therapy for treatment of atherosclerosis
CN106146670B (zh) 2015-04-24 2019-01-15 宜明昂科生物医药技术（上海）有限公司 一种新的重组双功能融合蛋白及其制备和应用
JP6898303B2 (ja) 2015-08-07 2021-07-07 エーエルエックス オンコロジー インコーポレイテッド Ｓｉｒｐ−アルファドメインまたはそのバリアントを有する構築物
SG11201808465UA (en) 2016-04-14 2018-10-30 Ose Immunotherapeutics NEW ANTI-SIRPa ANTIBODIES AND THEIR THERAPEUTIC APPLICATIONS
JOP20190009A1 (ar) 2016-09-21 2019-01-27 Alx Oncology Inc أجسام مضادة ضد بروتين ألفا منظم للإشارات وطرق استخدامها
US11613564B2 (en) 2019-05-31 2023-03-28 ALX Oncology Inc. Methods of treating cancer
JP7561775B2 (ja) 2019-06-07 2024-10-04 エーエルエックス オンコロジー インコーポレイテッド 血清学的アッセイにおいてｃｄ４７に結合する薬物の干渉を低減するための方法及び試薬
JP7713447B2 (ja) 2019-11-27 2025-07-25 エーエルエックス オンコロジー インコーポレイテッド がんを治療するための組み合わせ療法
CA3178157A1 (en) 2020-06-01 2021-12-09 Jaume Pons Combination therapies comprising a hypomethylation agent for treating cancer
WO2022010806A1 (en) 2020-07-06 2022-01-13 ALX Oncology Inc. Methods for reducing the interference of drugs that bind therapeutic targets expressed on blood cells in serological assays
US20220196651A1 (en) 2020-12-06 2022-06-23 ALX Oncology Inc. Multimers for reducing the interference of drugs that bind cd47 in serological assays
JP2024520902A (ja) 2021-05-13 2024-05-27 エーエルエックス オンコロジー インコーポレイテッド がんを治療するための併用療法

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100239579A1 (en) * 2006-05-15 2010-09-23 Viral Logic Systems Technology Corp. CD47 Related Compositions and Methods for Treating Immunological Diseases and Disorders
WO2010070047A1 (en) 2008-12-19 2010-06-24 Novartis Ag Soluble polypeptides for use in treating autoimmune and inflammatory disorders
WO2012142515A2 (en) * 2011-04-13 2012-10-18 Bristol-Myers Squibb Company Fc fusion proteins comprising novel linkers or arrangements
WO2013109752A1 (en) 2012-01-17 2013-07-25 The Board Of Trustees Of The Leland Stanford Junior University High affinity sirp-alpha reagents
WO2016023040A1 (en) * 2014-08-08 2016-02-11 Alexo Therapeutics International Sirp-alpha variant constructs and uses thereof

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
"UniProt", Database accession no. P02768
BORROK ET AL.: "Revisiting the role of glycosylation in the structure of human IgG Fc", ACS CHEMICAL BIOLOGY, vol. 7, 21 September 2012 (2012-09-21), pages 1596 - 1602, XP055363306 *
KABAT: "Sequences of Proteins of Immunological Interest", 1991, PUBLIC HEALTH SERVICE, NATIONAL INSTITUTES OF HEALTH
LEE ET AL.: "Novel structural determinants on SIRP that mediate binding to CD 47", THE JOURNAL OF IMMUNOLOGY, vol. 179, 2007, pages 7741 - 7750, XP055081468 *

Cited By (136)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10696730B2 (en) 2015-08-07 2020-06-30 ALX Oncology Inc. Constructs having a SIRP-alpha domain or variant thereof
US10259859B2 (en) 2015-08-07 2019-04-16 ALX Oncology Inc. Constructs having a SIRP-α domain or variant thereof
US11208459B2 (en) 2015-08-07 2021-12-28 ALX Oncology Inc. Constructs having a SIRP-alpha domain or variant thereof
US11639376B2 (en) 2015-08-07 2023-05-02 ALX Oncology Inc. Constructs having a SIRP-α domain or variant thereof
EP3341015B1 (en) 2015-08-26 2021-07-28 The Board of Trustees of the Leland Stanford Junior University Enhanced depletion of targeted cells with cd47 blockade and an immune costimulatory agonist
US12030944B2 (en) 2015-08-26 2024-07-09 The Board Of Trustees Of The Leland Stanford Junior University Enhanced depletion of targeted cells with CD47 blockade and an immune costimulatory agonist
EP3341015B2 (en) † 2015-08-26 2023-12-27 The Board of Trustees of the Leland Stanford Junior University Enhanced depletion of targeted cells with cd47 blockade and an immune costimulatory agonist
US11608377B2 (en) 2015-08-26 2023-03-21 The Board Of Trustees Of The Leland Stanford Junior University Enhanced depletion of targeted cells with CD47 blockade and an immune costimulatory agonist
US12060423B2 (en) 2016-01-11 2024-08-13 Forty Seven, Inc. Humanized, mouse or chimeric anti-CD47 monoclonal antibodies
US10927173B2 (en) 2016-01-11 2021-02-23 Forty Seven, Inc. Humanized, mouse or chimeric anti-CD47 monoclonal antibodies
US11643461B2 (en) 2016-01-11 2023-05-09 Forty Seven, Inc. Humanized, mouse or chimeric anti-CD47 monoclonal antibodies
US12358964B2 (en) 2016-08-10 2025-07-15 Ajou University Industry-Academic Cooperation Foundation Heterodimeric Fc-fused cytokine and pharmaceutical composition comprising the same
US11692019B2 (en) 2016-08-10 2023-07-04 Ajou University Industry-Academic Cooperation Foundation Heterodimeric Fc-fused cytokine and pharmaceutical composition comprising the same
US11078249B2 (en) 2016-08-10 2021-08-03 Ajou University Industry-Academic Cooperation Foundation Heterodimeric Fc-fused cytokine and pharmaceutical composition comprising the same
US10696722B2 (en) 2016-08-10 2020-06-30 Ajou University Industry-Academic Cooperation Foundation Heterodimeric Fc-fused cytokine and pharmaceutical composition comprising the same
US11702458B2 (en) 2017-01-05 2023-07-18 Kahr Medical Ltd. PD1-41BBL fusion protein and methods of use thereof
US11130796B2 (en) 2017-01-05 2021-09-28 Kahr Medical Ltd. SIRPalpha-41BBL fusion protein and methods of use thereof
US11566060B2 (en) 2017-01-05 2023-01-31 Kahr Medical Ltd. PD1-CD70 fusion protein and methods of use thereof
US12331098B2 (en) 2017-01-05 2025-06-17 Kahr Medical Ltd. SIRPalpha-41BBL fusion protein and methods of use thereof
US11897937B2 (en) 2017-01-05 2024-02-13 Kahr Medical Ltd. SIRPalpha-41BBL fusion protein and methods of use thereof
US11299530B2 (en) 2017-01-05 2022-04-12 Kahr Medical Ltd. SIRP alpha-CD70 fusion protein and methods of use thereof
JP7128195B2 (ja) 2017-02-27 2022-08-30 シャタック ラボ，インコーポレイテッド 細胞外ドメインベースのキメラタンパク質の製造方法及び使用方法
JP2022159510A (ja) * 2017-02-27 2022-10-17 シャタック ラボ，インコーポレイテッド 細胞外ドメインベースのキメラタンパク質の製造方法及び使用方法
JP2020508329A (ja) * 2017-02-27 2020-03-19 シャタック ラボ，インコーポレイテッド 細胞外ドメインベースのキメラタンパク質の製造方法及び使用方法
CN110958888A (zh) * 2017-03-28 2020-04-03 延龄草治疗公司 Cd47阻断疗法
EP3600424A4 (en) * 2017-03-28 2020-12-23 Trillium Therapeutics Inc. CD47 BLOCKED THERAPY
US12497456B2 (en) 2017-05-08 2025-12-16 Shanghai Jmt-Bio Technology Co., Ltd. Bispecific recombinant protein and use thereof
US11518810B2 (en) 2017-05-08 2022-12-06 Shanghai Jmt-Bio Technology Co., Ltd. Bispecific recombinant protein and use thereof
EP3623388A4 (en) * 2017-05-08 2021-08-04 Shanghai JMT-Bio Technology Co., Ltd. BISPECIFIC RECOMBINANT PROTEIN AND ITS USE
WO2019047885A1 (en) * 2017-09-07 2019-03-14 Dingfu Biotarget Co., Ltd. IMMUNOCONJUGATES COMPRISING ALPHA REGULATORY SIGNAL PROTEIN
US11529425B2 (en) 2017-09-07 2022-12-20 Dingfu Biotarget Co., Ltd. Immunoconjugates comprising signal regulatory protein alpha
CN111051350A (zh) * 2017-09-07 2020-04-21 苏州丁孚靶点生物技术有限公司 包含信号调节蛋白α的免疫缀合物
JP2021511817A (ja) * 2017-11-20 2021-05-13 泰州▲邁▼博太科▲薬▼▲業▼有限公司Ｔａｉｚｈｏｕ Ｍａｂｔｅｃｈ Ｐｈａｒｍａｃｅｕｔｉｃａｌ Ｃｏ．， Ｌｔｄ Ｃｄ４７とｐｄ−ｌ１を標的にする二重機能の融合タンパク質
US12110332B2 (en) 2017-11-20 2024-10-08 Taizhou Mabtech Pharmaceutical Co., Ltd. Bifunctional fusion protein targeting CD47 and PD-L1
JP7163399B2 (ja) 2017-11-20 2022-10-31 泰州▲邁▼博太科▲薬▼▲業▼有限公司 Ｃｄ４７とｐｄ－ｌ１を標的にする二重機能の融合タンパク質
JP2021505195A (ja) * 2017-12-04 2021-02-18 北京韓美薬品有限公司Ｂｅｉｊｉｎｇ Ｈａｎｍｉ Ｐｈａｒｍ． Ｃｏ．， Ｌｔｄ． 天然抗体様構造を有し、ヘテロ二量体形態の抗ｐｄ−ｌ１／抗ｃｄ４７二重特異性抗体、ならびにその製造方法
US11739151B2 (en) 2017-12-04 2023-08-29 Beijing Hanmi Pharmaceutical Co., Ltd. Anti-PD-L1/anti-CD47 bispecific antibody
JP7231641B2 (ja) 2017-12-04 2023-03-01 北京韓美薬品有限公司 天然抗体様構造を有し、ヘテロ二量体形態の抗ｐｄ－ｌ１／抗ｃｄ４７二重特異性抗体、ならびにその製造方法
CN109971716A (zh) * 2017-12-28 2019-07-05 上海细胞治疗研究院 自分泌cd47抗体的egfr特异性car-t细胞及其用途
CN109971716B (zh) * 2017-12-28 2023-08-01 上海细胞治疗研究院 自分泌cd47抗体的egfr特异性car-t细胞及其用途
US11891450B2 (en) 2018-02-12 2024-02-06 Forty Seven, Inc. Anti-CD47 agent-based treatment of CD20-positive cancer
WO2019157432A1 (en) 2018-02-12 2019-08-15 Forty Seven, Inc. Anti-cancer regimen using anti-cd47 and anti-cd20 antibodies
EP4129336A1 (en) 2018-02-12 2023-02-08 Forty Seven, Inc. Anti-cd47 agent-based treatment of cd20-positive cancer
CN111836647A (zh) * 2018-03-13 2020-10-27 延龄草治疗公司 Cd47阻断疗法和cd38抗体的组合
JP2021515779A (ja) * 2018-03-13 2021-06-24 トリリアム・セラピューティクス・インコーポレイテッドＴｒｉｌｌｉｕｍ Ｔｈｅｒａｐｅｕｔｉｃｓ Ｉｎｃ． Ｅｇｆｒ抗体によるｃｄ４７遮断療法における改善
CN111936167A (zh) * 2018-03-13 2020-11-13 延龄草治疗公司 Egfr抗体对cd47阻断疗法的改善
WO2019173903A1 (en) * 2018-03-13 2019-09-19 Trillium Therapeutics Inc. Improvements in cd47 blockade therapy by egfr antibody
JP2021517144A (ja) * 2018-03-13 2021-07-15 トリリアム・セラピューティクス・インコーポレイテッドＴｒｉｌｌｉｕｍ Ｔｈｅｒａｐｅｕｔｉｃｓ Ｉｎｃ． Ｃｄ４７遮断療法およびｃｄ３８抗体の組み合わせ
US11891423B2 (en) 2018-04-17 2024-02-06 Hangzhou Sumgen Biotech Co., Ltd. Fusion protein binding to CD47 protein and application thereof
EP3795595A4 (en) * 2018-04-17 2022-03-16 Hangzhou Sumgen Biotech Co., Ltd. FUSION PROTEIN THAT BINDS TO CD47 PROTEIN AND ITS USE
JP7756388B2 (ja) 2018-04-17 2025-10-20 杭州尚健生物技術有限公司 Ｃｄ４７タンパク質に結合する融合タンパク質およびその使用
JP2021520830A (ja) * 2018-04-17 2021-08-26 杭州尚健生物技術有限公司 Ｃｄ４７タンパク質に結合する融合タンパク質およびその使用
RU2787521C2 (ru) * 2018-04-17 2023-01-10 Ханчжоу Сумген Байотек Ко., Лтд. Связывание слитого белка с белком cd47 и его применение
WO2019234576A1 (en) * 2018-06-03 2019-12-12 Lamkap Bio Beta Ltd. Bispecific antibodies against ceacam5 and cd47
US11555071B2 (en) 2018-06-03 2023-01-17 Lamkap Bio Beta Ltd. Bispecific antibodies against CEACAM5 and CD47
US12286466B2 (en) 2018-07-11 2025-04-29 Kahr Medical Ltd. PD1-4-1BBL variant fusion protein and methods of use thereof
US12134638B2 (en) 2018-07-11 2024-11-05 Kahr Medical Ltd. SIRPalpha-4-1BBL variant fusion protein and methods of use thereof
WO2020012486A1 (en) 2018-07-11 2020-01-16 Kahr Medical Ltd. SIRPalpha-4-1BBL VARIANT FUSION PROTEIN AND METHODS OF USE THEREOF
CN112543773A (zh) * 2018-07-11 2021-03-23 卡尔医学有限公司 SIRPα-4-1BBL变体融合蛋白及其使用方法
CN112543773B (zh) * 2018-07-11 2024-05-31 卡尔医学有限公司 SIRPα-4-1BBL变体融合蛋白及其使用方法
WO2020047326A2 (en) 2018-08-31 2020-03-05 ALX Oncology Inc. Decoy polypeptides
JP2021534769A (ja) * 2018-08-31 2021-12-16 エーエルエックス オンコロジー インコーポレイテッド デコイポリペプチド
WO2020086758A1 (en) * 2018-10-23 2020-04-30 Dragonfly Therapeutics, Inc. Heterodimeric fc-fused proteins
US11787864B2 (en) 2018-10-23 2023-10-17 Dragonfly Therapeutics, Inc. Heterodimeric Fc-fused proteins
RU2800923C2 (ru) * 2018-11-14 2023-08-01 Ханчжоу Сумген Байотек Ко., Лтд. Слитый белок и его применение
US12304967B2 (en) 2018-11-14 2025-05-20 Hangzhou Sumgen Biotech Co., Ltd. Fusion protein and use thereof
WO2020200941A1 (en) 2019-03-29 2020-10-08 F. Hoffmann-La Roche Ag Spr-based binding assay for the functional analysis of multivalent molecules
WO2020243338A1 (en) * 2019-05-31 2020-12-03 ALX Oncology Inc. Methods of treating cancer with sirp alpha fc fusion in combination with an immune checkpoint inhibitor
US11613564B2 (en) 2019-05-31 2023-03-28 ALX Oncology Inc. Methods of treating cancer
EP3980051A4 (en) * 2019-06-05 2023-09-13 Asher Biotherapeutics, Inc. FUSIONS OF MUTANT INTERLEUKIN-2 POLYPEPTIDES WITH ANTIGEN-BINDING MOLECULES TO MODULATE THE FUNCTION OF IMMUNE CELLS
WO2020247820A1 (en) 2019-06-07 2020-12-10 ALX Oncology Inc. Methods and reagents for reducing the interference of drugs that bind cd47 in serological assays
WO2021011544A1 (en) 2019-07-16 2021-01-21 Gilead Sciences, Inc. Hiv vaccines and methods of making and using
WO2021076908A1 (en) 2019-10-18 2021-04-22 Forty Seven, Inc. Combination therapies for treating myelodysplastic syndromes and acute myeloid leukemia
EP4349413A2 (en) 2019-10-18 2024-04-10 Forty Seven, Inc. Combination therapies for treating myelodysplastic syndromes and acute myeloid leukemia
WO2021087064A1 (en) 2019-10-31 2021-05-06 Forty Seven, Inc. Anti-cd47 and anti-cd20 based treatment of blood cancer
WO2021108693A1 (en) 2019-11-27 2021-06-03 ALX Oncology Inc. Combination therapies for treating cancer
US20210154269A1 (en) * 2019-11-27 2021-05-27 ALX Oncology Inc. Combination therapies for treating cancer
WO2021130638A1 (en) 2019-12-24 2021-07-01 Carna Biosciences, Inc. Diacylglycerol kinase modulating compounds
EP4445902A2 (en) 2019-12-24 2024-10-16 Carna Biosciences, Inc. Diacylglycerol kinase modulating compounds
WO2021163064A2 (en) 2020-02-14 2021-08-19 Jounce Therapeutics, Inc. Antibodies and fusion proteins that bind to ccr8 and uses thereof
US11692038B2 (en) 2020-02-14 2023-07-04 Gilead Sciences, Inc. Antibodies that bind chemokine (C-C motif) receptor 8 (CCR8)
US12297282B2 (en) 2020-02-14 2025-05-13 Gilead Sciences, Inc. Nucleic acids encoding, and methods of producing, antibodies that bind human chemokine (C—C motif) receptor 8 (CCR8)
US12091681B2 (en) 2020-03-27 2024-09-17 Mendus B.V. Ex vivo use of modified cells of leukemic origin for enhancing the efficacy of adoptive cell therapy
WO2021247430A1 (en) 2020-06-01 2021-12-09 ALX Oncology Inc. Combination therapies comprising a hypomethylation agent for treating cancer
US20220401516A1 (en) * 2020-06-01 2022-12-22 ALX Oncology Inc. Combination therapies comprising a hypomethylation agent for treating cancer
US12343377B2 (en) 2020-06-01 2025-07-01 ALX Oncology Inc. Combination therapies comprising a hypomethylation agent for treating cancer
EP4155318A4 (en) * 2020-06-30 2024-06-05 Harbour Biomed US, Inc. BISPECIFIC ANTIBODY AND USE THEREOF
US12364758B2 (en) 2020-06-30 2025-07-22 Mendus B.V. Use of leukemia-derived cells in ovarian cancer vaccines
WO2022010806A1 (en) 2020-07-06 2022-01-13 ALX Oncology Inc. Methods for reducing the interference of drugs that bind therapeutic targets expressed on blood cells in serological assays
EP4190803A4 (en) * 2020-07-30 2024-08-21 Sunshine Guojian Pharmaceutical (Shanghai) Co., Ltd. SIRPa-FC FUSION PROTEIN
US12024559B2 (en) 2020-10-23 2024-07-02 Asher Biotherapeutics, Inc. Fusions with CD8 antigen binding molecules for modulating immune cell function
WO2022120286A1 (en) 2020-12-06 2022-06-09 ALX Oncology Inc. Multimers for reducing the interference of drugs that bind cd47 in serological assays
WO2022190058A1 (en) 2021-03-12 2022-09-15 Dcprime B.V. Methods of vaccination and use of cd47 blockade
WO2022221304A1 (en) 2021-04-14 2022-10-20 Gilead Sciences, Inc. CO-INHIBITION OF CD47/SIRPα BINDING AND NEDD8-ACTIVATING ENZYME E1 REGULATORY SUBUNIT FOR THE TREATMENT OF CANCER
US12098214B2 (en) 2021-05-13 2024-09-24 ALX Oncology Inc. Combination therapies for treating cancer
WO2022241157A1 (en) 2021-05-13 2022-11-17 ALX Oncology Inc. Combination therapies for treating cancer
US12247072B2 (en) 2021-05-19 2025-03-11 Asher Biotherapeutics, Inc. IL-21 polypeptides and targeted constructs
US12122827B2 (en) 2021-05-19 2024-10-22 Asher Biotherapeutics, Inc. IL-21 polypeptides and targeted constructs
WO2022271684A1 (en) 2021-06-23 2022-12-29 Gilead Sciences, Inc. Diacylglyercol kinase modulating compounds
WO2022271650A1 (en) 2021-06-23 2022-12-29 Gilead Sciences, Inc. Diacylglyercol kinase modulating compounds
WO2022271659A1 (en) 2021-06-23 2022-12-29 Gilead Sciences, Inc. Diacylglyercol kinase modulating compounds
WO2022271677A1 (en) 2021-06-23 2022-12-29 Gilead Sciences, Inc. Diacylglyercol kinase modulating compounds
WO2023076983A1 (en) 2021-10-28 2023-05-04 Gilead Sciences, Inc. Pyridizin-3(2h)-one derivatives
WO2023077030A1 (en) 2021-10-29 2023-05-04 Gilead Sciences, Inc. Cd73 compounds
US12447195B2 (en) 2021-12-21 2025-10-21 Fbd Biologics Limited Engineered SIRPα variants and methods of use thereof
EP4319796A4 (en) * 2021-12-21 2025-07-23 Fbd Biologics Ltd MODIFIED SIRPalpha VARIANTS AND METHODS OF USE THEREOF
WO2023122615A1 (en) 2021-12-22 2023-06-29 Gilead Sciences, Inc. Ikaros zinc finger family degraders and uses thereof
WO2023122581A2 (en) 2021-12-22 2023-06-29 Gilead Sciences, Inc. Ikaros zinc finger family degraders and uses thereof
WO2023147418A1 (en) 2022-01-28 2023-08-03 Gilead Sciences, Inc. Parp7 inhibitors
WO2023154578A1 (en) 2022-02-14 2023-08-17 Sana Biotechnology, Inc. Methods of treating patients exhibiting a prior failed therapy with hypoimmunogenic cells
WO2023178181A1 (en) 2022-03-17 2023-09-21 Gilead Sciences, Inc. Ikaros zinc finger family degraders and uses thereof
EP4464703A2 (en) 2022-03-17 2024-11-20 Gilead Sciences, Inc. Ikaros zinc finger family degraders and uses thereof
EP4245756A1 (en) 2022-03-17 2023-09-20 Gilead Sciences, Inc. Ikaros zinc finger family degraders and uses thereof
WO2023183313A1 (en) 2022-03-22 2023-09-28 Sana Biotechnology, Inc. Engineering cells with a transgene in b2m or ciita locus and associated compositions and methods
WO2023183890A1 (en) 2022-03-24 2023-09-28 Bitterroot Bio, Inc. Multivalent sirp-alpha fusion polypeptides
WO2023183892A1 (en) 2022-03-24 2023-09-28 Bitterroot Bio, Inc. Sirp-alpha fusion polypeptides with modified fc domains
WO2023183817A1 (en) 2022-03-24 2023-09-28 Gilead Sciences, Inc. Combination therapy for treating trop-2 expressing cancers
WO2023196784A1 (en) 2022-04-05 2023-10-12 Gilead Sciences, Inc. Combinations of antibody therapies for treating colorectal cancer
WO2023205719A1 (en) 2022-04-21 2023-10-26 Gilead Sciences, Inc. Kras g12d modulating compounds
WO2023218378A1 (en) 2022-05-11 2023-11-16 Daiichi Sankyo Company, Limited Combination of an antibody specific for a tumor antigen and a cd47 inhibitor
WO2023235754A1 (en) 2022-06-01 2023-12-07 ALX Oncology Inc. Combination therapies for treating urothelial carcinoma
WO2024006929A1 (en) 2022-07-01 2024-01-04 Gilead Sciences, Inc. Cd73 compounds
WO2024015741A1 (en) 2022-07-12 2024-01-18 Gilead Sciences, Inc. Hiv immunogenic polypeptides and vaccines and uses thereof
WO2024064668A1 (en) 2022-09-21 2024-03-28 Gilead Sciences, Inc. FOCAL IONIZING RADIATION AND CD47/SIRPα DISRUPTION ANTICANCER COMBINATION THERAPY
WO2024137852A1 (en) 2022-12-22 2024-06-27 Gilead Sciences, Inc. Prmt5 inhibitors and uses thereof
WO2024215754A1 (en) 2023-04-11 2024-10-17 Gilead Sciences, Inc. Kras modulating compounds
WO2024220917A1 (en) 2023-04-21 2024-10-24 Gilead Sciences, Inc. Prmt5 inhibitors and uses thereof
US20240400691A1 (en) * 2023-05-31 2024-12-05 Fbd Biologics Limited Cd47/pd-l1-targeting protein complex and methods of use thereof
WO2025006720A1 (en) 2023-06-30 2025-01-02 Gilead Sciences, Inc. Kras modulating compounds
WO2025024811A1 (en) 2023-07-26 2025-01-30 Gilead Sciences, Inc. Parp7 inhibitors
WO2025024663A1 (en) 2023-07-26 2025-01-30 Gilead Sciences, Inc. Parp7 inhibitors
WO2025054347A1 (en) 2023-09-08 2025-03-13 Gilead Sciences, Inc. Kras g12d modulating compounds
WO2025054530A1 (en) 2023-09-08 2025-03-13 Gilead Sciences, Inc. Pyrimidine-containing polycyclic derivatives as kras g12d modulating compounds
WO2025096589A1 (en) 2023-11-03 2025-05-08 Gilead Sciences, Inc. Prmt5 inhibitors and uses thereof
WO2025137640A1 (en) 2023-12-22 2025-06-26 Gilead Sciences, Inc. Azaspiro wrn inhibitors
WO2025245003A1 (en) 2024-05-21 2025-11-27 Gilead Sciences, Inc. Prmt5 inhibitors and uses thereof

Also Published As

Publication number Publication date
AU2016304794B2 (en) 2021-07-15
AU2021218004A1 (en) 2021-09-09
RU2020143675A (ru) 2021-05-26
IL294679B2 (en) 2023-10-01
RU2740672C2 (ru) 2021-01-19
AU2016304794A1 (en) 2018-02-01
KR20250133806A (ko) 2025-09-08
KR20180028548A (ko) 2018-03-16
US20170107270A1 (en) 2017-04-20
JP2018525382A (ja) 2018-09-06
GEP20217326B (en) 2021-11-25
JP2021063082A (ja) 2021-04-22
US11639376B2 (en) 2023-05-02
ZA202001863B (en) 2023-12-20
RU2018108103A (ru) 2019-09-09
US20190169266A1 (en) 2019-06-06
AU2021218004B2 (en) 2023-06-15
NZ778013A (en) 2025-02-28
PH12018500269B1 (en) 2023-10-20
US10696730B2 (en) 2020-06-30
CA2993835A1 (en) 2017-02-16
CN108350048B (zh) 2024-02-09
MX2022010803A (es) 2022-09-27
CN108350048A (zh) 2018-07-31
MX2018001428A (es) 2018-09-05
DK3331902T3 (da) 2021-07-26
CL2020000530A1 (es) 2020-07-10
JP6898303B2 (ja) 2021-07-07
CO2018002471A2 (es) 2018-05-31
EP3331902A1 (en) 2018-06-13
PE20180778A1 (es) 2018-05-07
UA125638C2 (uk) 2022-05-11
ZA202000743B (en) 2022-07-27
IL256989B (en) 2022-08-01
MX395275B (es) 2025-03-25
JP7591917B2 (ja) 2024-11-29
IL294679A (en) 2022-09-01
EP3331902B1 (en) 2021-04-28
GEP20227447B (en) 2022-12-12
EP3913051A1 (en) 2021-11-24
SG10201912905VA (en) 2020-02-27
JP2025069164A (ja) 2025-04-30
ES2881771T3 (es) 2021-11-30
NZ738950A (en) 2023-03-31
MY185014A (en) 2021-04-30
CL2018000340A1 (es) 2018-05-11
US10259859B2 (en) 2019-04-16
IL302491A (en) 2023-06-01
LT3331902T (lt) 2021-08-10
HRP20211167T1 (hr) 2021-10-15
PL3331902T3 (pl) 2021-11-22
RU2018108103A3 (enExample) 2019-12-20
UA130083C2 (uk) 2025-11-05
CL2020000531A1 (es) 2020-07-10
US20200239543A1 (en) 2020-07-30
US20240343778A1 (en) 2024-10-17
IL294679B1 (en) 2023-06-01
IL256989A (en) 2018-03-29
RS62151B1 (sr) 2021-08-31
BR112018001353A2 (pt) 2018-09-11
EP3331902A4 (en) 2019-04-03
PT3331902T (pt) 2021-07-26
ZA201800857B (en) 2020-10-28
SG10202101909RA (en) 2021-04-29
US11208459B2 (en) 2021-12-28
PH12018500269A1 (en) 2018-08-29
US20210070838A1 (en) 2021-03-11
KR102855748B1 (ko) 2025-09-09
HK1257372A1 (zh) 2019-10-18
SA518390881B1 (ar) 2021-11-14
CN117964732A (zh) 2024-05-03
CN117777267A (zh) 2024-03-29
RU2020143675A3 (enExample) 2021-10-29
SMT202100424T1 (it) 2021-09-14
HUE055139T2 (hu) 2021-11-29
SI3331902T1 (sl) 2021-09-30
CY1124372T1 (el) 2022-07-22
JP2023061969A (ja) 2023-05-02

Similar Documents

Publication Publication Date Title
US20240343778A1 (en) Constructs having a sirp-alpha domain or variant thereof
AU2015303135B2 (en) SIRP-alpha immunoglobulin fusion proteins
EP2927321B1 (en) Ch3 domain variant pair inducing formation of heterodimer of heavy chain constant region of antibody at high efficiency, method for preparing same, and use thereof
CA3116188A1 (en) Pd-1 targeted il-15/il-15ralpha fc fusion proteins and uses in combination therapies thereof
TW202221015A (zh) 單一及雙靶定配體誘導之t細胞銜接體組合物
WO2022150792A1 (en) Indinavir based chemical dimerization t cell engager compositions
HK40064097A (en) Constructs having a sirp-alpha domain or variant thereof
BR112018001353B1 (pt) Polipeptídeo compreendendo uma variante d1 da proteína reguladora de sinal a (sirp-a) e uma variante fc, seu método de produção e seus usos, célula hospedeira, e composição farmacêutica
HK1257372B (zh) 具有SIRP-α结构域或其变体的构建体

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: 16835723

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 256989

Country of ref document: IL

WWE Wipo information: entry into national phase

Ref document number: 122021007097

Country of ref document: BR

ENP Entry into the national phase

Ref document number: 2993835

Country of ref document: CA

ENP Entry into the national phase

Ref document number: 2016304794

Country of ref document: AU

Date of ref document: 20160805

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: MX/A/2018/001428

Country of ref document: MX

WWE Wipo information: entry into national phase

Ref document number: 000186-2018

Country of ref document: PE

Ref document number: 11201800903Q

Country of ref document: SG

WWE Wipo information: entry into national phase

Ref document number: 12018500269

Country of ref document: PH

ENP Entry into the national phase

Ref document number: 2018506281

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 14720

Country of ref document: GE

ENP Entry into the national phase

Ref document number: 20187006473

Country of ref document: KR

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: NC2018/0002471

Country of ref document: CO

Ref document number: NC2020/0001932

Country of ref document: CO

WWE Wipo information: entry into national phase

Ref document number: 2018108103

Country of ref document: RU

Ref document number: 2016835723

Country of ref document: EP

REG Reference to national code

Ref country code: BR

Ref legal event code: B01A

Ref document number: 112018001353

Country of ref document: BR

WWE Wipo information: entry into national phase

Ref document number: 201680057835.8

Country of ref document: CN

WWP Wipo information: published in national office

Ref document number: NC2018/0002471

Country of ref document: CO

ENP Entry into the national phase

Ref document number: 112018001353

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20180123

WWG Wipo information: grant in national office

Ref document number: NC2018/0002471

Country of ref document: CO

WWE Wipo information: entry into national phase

Ref document number: 15294/1

Country of ref document: GE

WWW Wipo information: withdrawn in national office

Ref document number: 302491

Country of ref document: IL