WO2020072395A1 - Combination cell-based therapies - Google Patents
Combination cell-based therapiesInfo
- Publication number
- WO2020072395A1 WO2020072395A1 PCT/US2019/053925 US2019053925W WO2020072395A1 WO 2020072395 A1 WO2020072395 A1 WO 2020072395A1 US 2019053925 W US2019053925 W US 2019053925W WO 2020072395 A1 WO2020072395 A1 WO 2020072395A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cells
- cell
- fusion protein
- tumor
- patient
- Prior art date
Links
- 238000002560 therapeutic procedure Methods 0.000 title description 57
- 210000004027 cell Anatomy 0.000 claims abstract description 576
- 210000001744 T-lymphocyte Anatomy 0.000 claims abstract description 185
- 238000000034 method Methods 0.000 claims abstract description 95
- 229960005486 vaccine Drugs 0.000 claims abstract description 77
- 238000011282 treatment Methods 0.000 claims abstract description 48
- 206010028980 Neoplasm Diseases 0.000 claims description 189
- 108020001507 fusion proteins Proteins 0.000 claims description 128
- 102000037865 fusion proteins Human genes 0.000 claims description 128
- 230000000139 costimulatory effect Effects 0.000 claims description 102
- 108090000623 proteins and genes Proteins 0.000 claims description 85
- 241000282414 Homo sapiens Species 0.000 claims description 77
- 102000004169 proteins and genes Human genes 0.000 claims description 71
- 239000000427 antigen Substances 0.000 claims description 64
- 108091007433 antigens Proteins 0.000 claims description 63
- 102000036639 antigens Human genes 0.000 claims description 63
- 201000011510 cancer Diseases 0.000 claims description 63
- 239000013604 expression vector Substances 0.000 claims description 40
- 239000002773 nucleotide Substances 0.000 claims description 40
- 125000003729 nucleotide group Chemical group 0.000 claims description 40
- 230000004913 activation Effects 0.000 claims description 38
- 238000002255 vaccination Methods 0.000 claims description 36
- 239000003795 chemical substances by application Substances 0.000 claims description 27
- 210000004881 tumor cell Anatomy 0.000 claims description 26
- 208000015181 infectious disease Diseases 0.000 claims description 25
- 101100454807 Caenorhabditis elegans lgg-1 gene Proteins 0.000 claims description 16
- 230000001965 increasing effect Effects 0.000 claims description 15
- 230000035755 proliferation Effects 0.000 claims description 15
- 101150013553 CD40 gene Proteins 0.000 claims description 13
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 claims description 13
- 241000700605 Viruses Species 0.000 claims description 13
- 101100454808 Caenorhabditis elegans lgg-2 gene Proteins 0.000 claims description 12
- 101100217502 Caenorhabditis elegans lgg-3 gene Proteins 0.000 claims description 11
- 101000679903 Homo sapiens Tumor necrosis factor receptor superfamily member 25 Proteins 0.000 claims description 11
- 102100022203 Tumor necrosis factor receptor superfamily member 25 Human genes 0.000 claims description 11
- 102100027207 CD27 antigen Human genes 0.000 claims description 9
- 101000914511 Homo sapiens CD27 antigen Proteins 0.000 claims description 9
- 241000725303 Human immunodeficiency virus Species 0.000 claims description 9
- 230000001154 acute effect Effects 0.000 claims description 9
- 230000001506 immunosuppresive effect Effects 0.000 claims description 8
- 229960003301 nivolumab Drugs 0.000 claims description 8
- 229960002621 pembrolizumab Drugs 0.000 claims description 6
- 241000711549 Hepacivirus C Species 0.000 claims description 5
- 208000037581 Persistent Infection Diseases 0.000 claims description 5
- 201000004792 malaria Diseases 0.000 claims description 5
- 241000700721 Hepatitis B virus Species 0.000 claims description 4
- 229950010773 pidilizumab Drugs 0.000 claims description 4
- 229940121420 cemiplimab Drugs 0.000 claims description 3
- 230000001717 pathogenic effect Effects 0.000 claims description 3
- 229950007213 spartalizumab Drugs 0.000 claims description 3
- 101000801234 Homo sapiens Tumor necrosis factor receptor superfamily member 18 Proteins 0.000 claims description 2
- 102100033728 Tumor necrosis factor receptor superfamily member 18 Human genes 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims description 2
- 101000946843 Homo sapiens T-cell surface glycoprotein CD8 alpha chain Proteins 0.000 description 68
- 102100034922 T-cell surface glycoprotein CD8 alpha chain Human genes 0.000 description 68
- 235000018102 proteins Nutrition 0.000 description 63
- 230000014509 gene expression Effects 0.000 description 62
- 241000699666 Mus <mouse, genus> Species 0.000 description 45
- 108090000765 processed proteins & peptides Proteins 0.000 description 41
- 241000699670 Mus sp. Species 0.000 description 39
- 241001465754 Metazoa Species 0.000 description 35
- -1 4-1 BB Proteins 0.000 description 33
- 230000004044 response Effects 0.000 description 33
- 230000003248 secreting effect Effects 0.000 description 33
- 230000004927 fusion Effects 0.000 description 31
- 230000004614 tumor growth Effects 0.000 description 30
- 101710089372 Programmed cell death protein 1 Proteins 0.000 description 27
- 102100040678 Programmed cell death protein 1 Human genes 0.000 description 27
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 26
- 125000003275 alpha amino acid group Chemical group 0.000 description 25
- 102000004473 OX40 Ligand Human genes 0.000 description 24
- 108010042215 OX40 Ligand Proteins 0.000 description 24
- 102000004196 processed proteins & peptides Human genes 0.000 description 24
- 210000003171 tumor-infiltrating lymphocyte Anatomy 0.000 description 24
- 235000001014 amino acid Nutrition 0.000 description 23
- 210000001266 CD8-positive T-lymphocyte Anatomy 0.000 description 22
- 238000000684 flow cytometry Methods 0.000 description 22
- 229940024606 amino acid Drugs 0.000 description 21
- 230000028993 immune response Effects 0.000 description 21
- 239000013598 vector Substances 0.000 description 21
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 description 20
- 108050005493 CD3 protein, epsilon/gamma/delta subunit Proteins 0.000 description 20
- 230000028327 secretion Effects 0.000 description 20
- 102100040004 Gamma-glutamylcyclotransferase Human genes 0.000 description 19
- 101000886680 Homo sapiens Gamma-glutamylcyclotransferase Proteins 0.000 description 19
- 101000716102 Homo sapiens T-cell surface glycoprotein CD4 Proteins 0.000 description 19
- 102100036011 T-cell surface glycoprotein CD4 Human genes 0.000 description 19
- 150000001413 amino acids Chemical class 0.000 description 19
- 239000000203 mixture Substances 0.000 description 19
- 239000000556 agonist Substances 0.000 description 18
- 210000004369 blood Anatomy 0.000 description 18
- 239000008280 blood Substances 0.000 description 18
- 239000012634 fragment Substances 0.000 description 18
- 210000000952 spleen Anatomy 0.000 description 16
- 238000006467 substitution reaction Methods 0.000 description 16
- 210000001519 tissue Anatomy 0.000 description 16
- 102100021317 Inducible T-cell costimulator Human genes 0.000 description 15
- 101710205775 Inducible T-cell costimulator Proteins 0.000 description 15
- 238000003114 enzyme-linked immunosorbent spot assay Methods 0.000 description 15
- 230000001939 inductive effect Effects 0.000 description 15
- 230000005764 inhibitory process Effects 0.000 description 15
- 238000002965 ELISA Methods 0.000 description 14
- 101000957437 Homo sapiens Mitochondrial carnitine/acylcarnitine carrier protein Proteins 0.000 description 13
- 102100038738 Mitochondrial carnitine/acylcarnitine carrier protein Human genes 0.000 description 13
- 238000004458 analytical method Methods 0.000 description 13
- 230000000259 anti-tumor effect Effects 0.000 description 13
- 230000000694 effects Effects 0.000 description 13
- 101000829958 Homo sapiens N-acetyllactosaminide beta-1,6-N-acetylglucosaminyl-transferase Proteins 0.000 description 12
- 102100023315 N-acetyllactosaminide beta-1,6-N-acetylglucosaminyl-transferase Human genes 0.000 description 12
- 208000036142 Viral infection Diseases 0.000 description 12
- 210000004962 mammalian cell Anatomy 0.000 description 12
- 150000007523 nucleic acids Chemical class 0.000 description 12
- 210000005259 peripheral blood Anatomy 0.000 description 12
- 239000011886 peripheral blood Substances 0.000 description 12
- 102100025230 2-amino-3-ketobutyrate coenzyme A ligase, mitochondrial Human genes 0.000 description 11
- 108010087522 Aeromonas hydrophilia lipase-acyltransferase Proteins 0.000 description 11
- 101000651036 Arabidopsis thaliana Galactolipid galactosyltransferase SFR2, chloroplastic Proteins 0.000 description 11
- 102100039635 Cancer/testis antigen 47A Human genes 0.000 description 11
- 108020004414 DNA Proteins 0.000 description 11
- 101000746249 Homo sapiens Cancer/testis antigen 47A Proteins 0.000 description 11
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 description 11
- 230000000735 allogeneic effect Effects 0.000 description 11
- 125000000539 amino acid group Chemical group 0.000 description 11
- 230000034994 death Effects 0.000 description 11
- 230000003053 immunization Effects 0.000 description 11
- 238000002649 immunization Methods 0.000 description 11
- 238000001727 in vivo Methods 0.000 description 11
- 230000009385 viral infection Effects 0.000 description 11
- 108010021064 CTLA-4 Antigen Proteins 0.000 description 10
- 229940045513 CTLA4 antagonist Drugs 0.000 description 10
- 102100039498 Cytotoxic T-lymphocyte protein 4 Human genes 0.000 description 10
- 101000724418 Homo sapiens Neutral amino acid transporter B(0) Proteins 0.000 description 10
- 108060003951 Immunoglobulin Proteins 0.000 description 10
- 102100028267 Neutral amino acid transporter B(0) Human genes 0.000 description 10
- 239000002671 adjuvant Substances 0.000 description 10
- 231100000371 dose-limiting toxicity Toxicity 0.000 description 10
- 102000018358 immunoglobulin Human genes 0.000 description 10
- 210000004988 splenocyte Anatomy 0.000 description 10
- 238000007619 statistical method Methods 0.000 description 10
- 239000006228 supernatant Substances 0.000 description 10
- JTTIOYHBNXDJOD-UHFFFAOYSA-N 2,4,6-triaminopyrimidine Chemical compound NC1=CC(N)=NC(N)=N1 JTTIOYHBNXDJOD-UHFFFAOYSA-N 0.000 description 9
- 102000004127 Cytokines Human genes 0.000 description 9
- 108090000695 Cytokines Proteins 0.000 description 9
- 241000282412 Homo Species 0.000 description 9
- 108091028043 Nucleic acid sequence Proteins 0.000 description 9
- 229920001213 Polysorbate 20 Polymers 0.000 description 9
- 239000006180 TBST buffer Substances 0.000 description 9
- 238000013461 design Methods 0.000 description 9
- 201000010099 disease Diseases 0.000 description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 9
- 201000001441 melanoma Diseases 0.000 description 9
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 9
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 9
- 229920001184 polypeptide Polymers 0.000 description 9
- 102000005962 receptors Human genes 0.000 description 9
- 108020003175 receptors Proteins 0.000 description 9
- 208000024891 symptom Diseases 0.000 description 9
- 238000012546 transfer Methods 0.000 description 9
- LKKMLIBUAXYLOY-UHFFFAOYSA-N 3-Amino-1-methyl-5H-pyrido[4,3-b]indole Chemical compound N1C2=CC=CC=C2C2=C1C=C(N)N=C2C LKKMLIBUAXYLOY-UHFFFAOYSA-N 0.000 description 8
- 102100030310 5,6-dihydroxyindole-2-carboxylic acid oxidase Human genes 0.000 description 8
- 101710163881 5,6-dihydroxyindole-2-carboxylic acid oxidase Proteins 0.000 description 8
- 102100031413 L-dopachrome tautomerase Human genes 0.000 description 8
- 101710093778 L-dopachrome tautomerase Proteins 0.000 description 8
- 241001529936 Murinae Species 0.000 description 8
- 101710173693 Short transient receptor potential channel 1 Proteins 0.000 description 8
- LVTKHGUGBGNBPL-UHFFFAOYSA-N Trp-P-1 Chemical compound N1C2=CC=CC=C2C2=C1C(C)=C(N)N=C2C LVTKHGUGBGNBPL-UHFFFAOYSA-N 0.000 description 8
- 239000003242 anti bacterial agent Substances 0.000 description 8
- 238000011161 development Methods 0.000 description 8
- 238000010790 dilution Methods 0.000 description 8
- 239000012895 dilution Substances 0.000 description 8
- 229940079593 drug Drugs 0.000 description 8
- 239000003814 drug Substances 0.000 description 8
- 239000012636 effector Substances 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 8
- 230000006870 function Effects 0.000 description 8
- 210000004185 liver Anatomy 0.000 description 8
- 201000005249 lung adenocarcinoma Diseases 0.000 description 8
- 230000008569 process Effects 0.000 description 8
- 230000001105 regulatory effect Effects 0.000 description 8
- 238000010186 staining Methods 0.000 description 8
- 230000000638 stimulation Effects 0.000 description 8
- 231100000419 toxicity Toxicity 0.000 description 8
- 230000001988 toxicity Effects 0.000 description 8
- 208000010507 Adenocarcinoma of Lung Diseases 0.000 description 7
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 7
- 101000830596 Homo sapiens Tumor necrosis factor ligand superfamily member 15 Proteins 0.000 description 7
- 102100022430 Melanocyte protein PMEL Human genes 0.000 description 7
- 102100024216 Programmed cell death 1 ligand 1 Human genes 0.000 description 7
- 101800001271 Surface protein Proteins 0.000 description 7
- 230000005867 T cell response Effects 0.000 description 7
- 102100024587 Tumor necrosis factor ligand superfamily member 15 Human genes 0.000 description 7
- 230000002411 adverse Effects 0.000 description 7
- 229940088710 antibiotic agent Drugs 0.000 description 7
- 230000008901 benefit Effects 0.000 description 7
- 238000011156 evaluation Methods 0.000 description 7
- 210000000987 immune system Anatomy 0.000 description 7
- 210000004698 lymphocyte Anatomy 0.000 description 7
- 108020004707 nucleic acids Proteins 0.000 description 7
- 102000039446 nucleic acids Human genes 0.000 description 7
- 238000000528 statistical test Methods 0.000 description 7
- 230000004083 survival effect Effects 0.000 description 7
- 230000001225 therapeutic effect Effects 0.000 description 7
- UZOVYGYOLBIAJR-UHFFFAOYSA-N 4-isocyanato-4'-methyldiphenylmethane Chemical compound C1=CC(C)=CC=C1CC1=CC=C(N=C=O)C=C1 UZOVYGYOLBIAJR-UHFFFAOYSA-N 0.000 description 6
- 108010074708 B7-H1 Antigen Proteins 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 6
- 238000011740 C57BL/6 mouse Methods 0.000 description 6
- 102100032912 CD44 antigen Human genes 0.000 description 6
- 241000282693 Cercopithecidae Species 0.000 description 6
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 description 6
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 6
- 241000124008 Mammalia Species 0.000 description 6
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 6
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 6
- 108091008874 T cell receptors Proteins 0.000 description 6
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 description 6
- 230000002159 abnormal effect Effects 0.000 description 6
- 230000002924 anti-infective effect Effects 0.000 description 6
- 210000000612 antigen-presenting cell Anatomy 0.000 description 6
- 230000000903 blocking effect Effects 0.000 description 6
- 210000000481 breast Anatomy 0.000 description 6
- 150000001875 compounds Chemical class 0.000 description 6
- 239000003937 drug carrier Substances 0.000 description 6
- 230000001900 immune effect Effects 0.000 description 6
- 238000011534 incubation Methods 0.000 description 6
- 230000007774 longterm Effects 0.000 description 6
- 230000035772 mutation Effects 0.000 description 6
- 244000045947 parasite Species 0.000 description 6
- 230000003612 virological effect Effects 0.000 description 6
- JVJGCCBAOOWGEO-RUTPOYCXSA-N (2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-4-amino-2-[[(2s,3s)-2-[[(2s,3s)-2-[[(2s)-2-azaniumyl-3-hydroxypropanoyl]amino]-3-methylpentanoyl]amino]-3-methylpentanoyl]amino]-4-oxobutanoyl]amino]-3-phenylpropanoyl]amino]-4-carboxylatobutanoyl]amino]-6-azaniumy Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O)CC1=CC=CC=C1 JVJGCCBAOOWGEO-RUTPOYCXSA-N 0.000 description 5
- 206010006187 Breast cancer Diseases 0.000 description 5
- 210000004366 CD4-positive T-lymphocyte Anatomy 0.000 description 5
- 102100040870 Glycine amidinotransferase, mitochondrial Human genes 0.000 description 5
- 239000012981 Hank's balanced salt solution Substances 0.000 description 5
- 108010004889 Heat-Shock Proteins Proteins 0.000 description 5
- 102000002812 Heat-Shock Proteins Human genes 0.000 description 5
- 101000893303 Homo sapiens Glycine amidinotransferase, mitochondrial Proteins 0.000 description 5
- 101000814512 Homo sapiens X antigen family member 1 Proteins 0.000 description 5
- 241000829100 Macaca mulatta polyomavirus 1 Species 0.000 description 5
- 108010076504 Protein Sorting Signals Proteins 0.000 description 5
- 101710173694 Short transient receptor potential channel 2 Proteins 0.000 description 5
- 102100039490 X antigen family member 1 Human genes 0.000 description 5
- 230000000890 antigenic effect Effects 0.000 description 5
- 230000005975 antitumor immune response Effects 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 5
- 229940030156 cell vaccine Drugs 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 230000001684 chronic effect Effects 0.000 description 5
- 210000001072 colon Anatomy 0.000 description 5
- 239000002299 complementary DNA Substances 0.000 description 5
- 125000000151 cysteine group Chemical class N[C@@H](CS)C(=O)* 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 210000003292 kidney cell Anatomy 0.000 description 5
- 230000001855 preneoplastic effect Effects 0.000 description 5
- 230000010076 replication Effects 0.000 description 5
- FVFVNNKYKYZTJU-UHFFFAOYSA-N 6-chloro-1,3,5-triazine-2,4-diamine Chemical compound NC1=NC(N)=NC(Cl)=N1 FVFVNNKYKYZTJU-UHFFFAOYSA-N 0.000 description 4
- 208000035143 Bacterial infection Diseases 0.000 description 4
- 208000026310 Breast neoplasm Diseases 0.000 description 4
- 108091026890 Coding region Proteins 0.000 description 4
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 4
- 238000011510 Elispot assay Methods 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 4
- 102100036304 G antigen 12B/C/D/E Human genes 0.000 description 4
- 208000012766 Growth delay Diseases 0.000 description 4
- 101000934356 Homo sapiens CD70 antigen Proteins 0.000 description 4
- 101001074834 Homo sapiens G antigen 12B/C/D/E Proteins 0.000 description 4
- 229940076838 Immune checkpoint inhibitor Drugs 0.000 description 4
- 206010062016 Immunosuppression Diseases 0.000 description 4
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 4
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 4
- 206010025323 Lymphomas Diseases 0.000 description 4
- 206010027476 Metastases Diseases 0.000 description 4
- 208000030852 Parasitic disease Diseases 0.000 description 4
- 206010060862 Prostate cancer Diseases 0.000 description 4
- 241000700159 Rattus Species 0.000 description 4
- 206010039491 Sarcoma Diseases 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 230000020385 T cell costimulation Effects 0.000 description 4
- 238000013459 approach Methods 0.000 description 4
- 208000022362 bacterial infectious disease Diseases 0.000 description 4
- 230000004071 biological effect Effects 0.000 description 4
- 210000004556 brain Anatomy 0.000 description 4
- 238000004364 calculation method Methods 0.000 description 4
- 230000010261 cell growth Effects 0.000 description 4
- 230000001413 cellular effect Effects 0.000 description 4
- 238000002512 chemotherapy Methods 0.000 description 4
- 238000011284 combination treatment Methods 0.000 description 4
- 230000002596 correlated effect Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 231100000673 dose–response relationship Toxicity 0.000 description 4
- 210000003743 erythrocyte Anatomy 0.000 description 4
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 4
- 238000009472 formulation Methods 0.000 description 4
- 239000012274 immune-checkpoint protein inhibitor Substances 0.000 description 4
- 230000002163 immunogen Effects 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 230000003834 intracellular effect Effects 0.000 description 4
- UWKQSNNFCGGAFS-XIFFEERXSA-N irinotecan Chemical compound C1=C2C(CC)=C3CN(C(C4=C([C@@](C(=O)OC4)(O)CC)C=4)=O)C=4C3=NC2=CC=C1OC(=O)N(CC1)CCC1N1CCCCC1 UWKQSNNFCGGAFS-XIFFEERXSA-N 0.000 description 4
- 239000003446 ligand Substances 0.000 description 4
- 230000000670 limiting effect Effects 0.000 description 4
- 201000005202 lung cancer Diseases 0.000 description 4
- 208000020816 lung neoplasm Diseases 0.000 description 4
- 239000006166 lysate Substances 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000003550 marker Substances 0.000 description 4
- 229960000485 methotrexate Drugs 0.000 description 4
- 210000000056 organ Anatomy 0.000 description 4
- 239000008188 pellet Substances 0.000 description 4
- 229920000136 polysorbate Polymers 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 230000012743 protein tagging Effects 0.000 description 4
- 238000013207 serial dilution Methods 0.000 description 4
- 230000011664 signaling Effects 0.000 description 4
- 241000894007 species Species 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 4
- WYWHKKSPHMUBEB-UHFFFAOYSA-N tioguanine Chemical compound N1C(N)=NC(=S)C2=C1N=CN2 WYWHKKSPHMUBEB-UHFFFAOYSA-N 0.000 description 4
- 230000009261 transgenic effect Effects 0.000 description 4
- 241000701161 unidentified adenovirus Species 0.000 description 4
- FUOOLUPWFVMBKG-UHFFFAOYSA-N 2-Aminoisobutyric acid Chemical compound CC(C)(N)C(O)=O FUOOLUPWFVMBKG-UHFFFAOYSA-N 0.000 description 3
- ALYNCZNDIQEVRV-UHFFFAOYSA-N 4-aminobenzoic acid Chemical compound NC1=CC=C(C(O)=O)C=C1 ALYNCZNDIQEVRV-UHFFFAOYSA-N 0.000 description 3
- VVIAGPKUTFNRDU-UHFFFAOYSA-N 6S-folinic acid Natural products C1NC=2NC(N)=NC(=O)C=2N(C=O)C1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 VVIAGPKUTFNRDU-UHFFFAOYSA-N 0.000 description 3
- 102000010735 Adenomatous polyposis coli protein Human genes 0.000 description 3
- 108010038310 Adenomatous polyposis coli protein Proteins 0.000 description 3
- 241000606125 Bacteroides Species 0.000 description 3
- 241000283690 Bos taurus Species 0.000 description 3
- 241000701822 Bovine papillomavirus Species 0.000 description 3
- 102100024263 CD160 antigen Human genes 0.000 description 3
- 102100025221 CD70 antigen Human genes 0.000 description 3
- 102100025570 Cancer/testis antigen 1 Human genes 0.000 description 3
- 241000283707 Capra Species 0.000 description 3
- 201000009030 Carcinoma Diseases 0.000 description 3
- 206010009944 Colon cancer Diseases 0.000 description 3
- 208000035473 Communicable disease Diseases 0.000 description 3
- 241000701022 Cytomegalovirus Species 0.000 description 3
- 206010061818 Disease progression Diseases 0.000 description 3
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 3
- 101000761938 Homo sapiens CD160 antigen Proteins 0.000 description 3
- 101000856237 Homo sapiens Cancer/testis antigen 1 Proteins 0.000 description 3
- 101000971533 Homo sapiens Killer cell lectin-like receptor subfamily G member 1 Proteins 0.000 description 3
- 101000845189 Homo sapiens Testis-specific Y-encoded protein 1 Proteins 0.000 description 3
- 101000764263 Homo sapiens Tumor necrosis factor ligand superfamily member 4 Proteins 0.000 description 3
- 108010074328 Interferon-gamma Proteins 0.000 description 3
- 208000008839 Kidney Neoplasms Diseases 0.000 description 3
- 102100021457 Killer cell lectin-like receptor subfamily G member 1 Human genes 0.000 description 3
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 3
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 3
- 101100407308 Mus musculus Pdcd1lg2 gene Proteins 0.000 description 3
- 229930012538 Paclitaxel Natural products 0.000 description 3
- 241001494479 Pecora Species 0.000 description 3
- 108700030875 Programmed Cell Death 1 Ligand 2 Proteins 0.000 description 3
- 102100024213 Programmed cell death 1 ligand 2 Human genes 0.000 description 3
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 3
- 241000700584 Simplexvirus Species 0.000 description 3
- 206010041067 Small cell lung cancer Diseases 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 102100031283 Testis-specific Y-encoded protein 1 Human genes 0.000 description 3
- 108060008683 Tumor Necrosis Factor Receptor Proteins 0.000 description 3
- 102100022153 Tumor necrosis factor receptor superfamily member 4 Human genes 0.000 description 3
- 210000001015 abdomen Anatomy 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- QWCKQJZIFLGMSD-UHFFFAOYSA-N alpha-aminobutyric acid Chemical compound CCC(N)C(O)=O QWCKQJZIFLGMSD-UHFFFAOYSA-N 0.000 description 3
- 239000002246 antineoplastic agent Substances 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 229930195731 calicheamicin Natural products 0.000 description 3
- HXCHCVDVKSCDHU-LULTVBGHSA-N calicheamicin Chemical compound C1[C@H](OC)[C@@H](NCC)CO[C@H]1O[C@H]1[C@H](O[C@@H]2C\3=C(NC(=O)OC)C(=O)C[C@](C/3=C/CSSSC)(O)C#C\C=C/C#C2)O[C@H](C)[C@@H](NO[C@@H]2O[C@H](C)[C@@H](SC(=O)C=3C(=C(OC)C(O[C@H]4[C@@H]([C@H](OC)[C@@H](O)[C@H](C)O4)O)=C(I)C=3C)OC)[C@@H](O)C2)[C@@H]1O HXCHCVDVKSCDHU-LULTVBGHSA-N 0.000 description 3
- 208000029742 colonic neoplasm Diseases 0.000 description 3
- 238000002648 combination therapy Methods 0.000 description 3
- 235000018417 cysteine Nutrition 0.000 description 3
- 230000007423 decrease Effects 0.000 description 3
- 210000004443 dendritic cell Anatomy 0.000 description 3
- 230000001419 dependent effect Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 230000005750 disease progression Effects 0.000 description 3
- 238000010494 dissociation reaction Methods 0.000 description 3
- 230000005593 dissociations Effects 0.000 description 3
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 3
- 230000002708 enhancing effect Effects 0.000 description 3
- 238000013401 experimental design Methods 0.000 description 3
- 210000002950 fibroblast Anatomy 0.000 description 3
- 229960002949 fluorouracil Drugs 0.000 description 3
- 235000008191 folinic acid Nutrition 0.000 description 3
- 239000011672 folinic acid Substances 0.000 description 3
- VVIAGPKUTFNRDU-ABLWVSNPSA-N folinic acid Chemical compound C1NC=2NC(N)=NC(=O)C=2N(C=O)C1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 VVIAGPKUTFNRDU-ABLWVSNPSA-N 0.000 description 3
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 description 3
- 239000003862 glucocorticoid Substances 0.000 description 3
- 238000009169 immunotherapy Methods 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 230000000968 intestinal effect Effects 0.000 description 3
- 229960005386 ipilimumab Drugs 0.000 description 3
- 229960001691 leucovorin Drugs 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 210000005229 liver cell Anatomy 0.000 description 3
- 210000004072 lung Anatomy 0.000 description 3
- 238000002826 magnetic-activated cell sorting Methods 0.000 description 3
- 238000012423 maintenance Methods 0.000 description 3
- 108010082117 matrigel Proteins 0.000 description 3
- GLVAUDGFNGKCSF-UHFFFAOYSA-N mercaptopurine Chemical compound S=C1NC=NC2=C1NC=N2 GLVAUDGFNGKCSF-UHFFFAOYSA-N 0.000 description 3
- 230000009401 metastasis Effects 0.000 description 3
- 208000037819 metastatic cancer Diseases 0.000 description 3
- 208000011575 metastatic malignant neoplasm Diseases 0.000 description 3
- 206010061289 metastatic neoplasm Diseases 0.000 description 3
- 229960004857 mitomycin Drugs 0.000 description 3
- KKZJGLLVHKMTCM-UHFFFAOYSA-N mitoxantrone Chemical compound O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO KKZJGLLVHKMTCM-UHFFFAOYSA-N 0.000 description 3
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 3
- 229960001756 oxaliplatin Drugs 0.000 description 3
- DWAFYCQODLXJNR-BNTLRKBRSA-L oxaliplatin Chemical compound O1C(=O)C(=O)O[Pt]11N[C@@H]2CCCC[C@H]2N1 DWAFYCQODLXJNR-BNTLRKBRSA-L 0.000 description 3
- 229960001592 paclitaxel Drugs 0.000 description 3
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 230000037452 priming Effects 0.000 description 3
- 210000002307 prostate Anatomy 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 210000003289 regulatory T cell Anatomy 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000001177 retroviral effect Effects 0.000 description 3
- 208000011581 secondary neoplasm Diseases 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- 206010041823 squamous cell carcinoma Diseases 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 210000000130 stem cell Anatomy 0.000 description 3
- 230000009885 systemic effect Effects 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- 238000011269 treatment regimen Methods 0.000 description 3
- 102000003298 tumor necrosis factor receptor Human genes 0.000 description 3
- 241001430294 unidentified retrovirus Species 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- BKZOUCVNTCLNFF-IGXZVFLKSA-N (2s)-2-[(2r,3r,4s,5r,6s)-2-hydroxy-6-[(1s)-1-[(2s,5r,7s,8r,9s)-2-[(2r,5s)-5-[(2r,3s,4r,5r)-5-[(2s,3s,4s,5r,6s)-6-hydroxy-4-methoxy-3,5,6-trimethyloxan-2-yl]-4-methoxy-3-methyloxolan-2-yl]-5-methyloxolan-2-yl]-7-methoxy-2,8-dimethyl-1,10-dioxaspiro[4.5]dec Chemical compound O([C@@H]1[C@@H]2O[C@H]([C@@H](C)[C@H]2OC)[C@@]2(C)O[C@H](CC2)[C@@]2(C)O[C@]3(O[C@@H]([C@H](C)[C@@H](OC)C3)[C@@H](C)[C@@H]3[C@@H]([C@H](OC)[C@@H](C)[C@](O)([C@H](C)C(O)=O)O3)C)CC2)[C@](C)(O)[C@H](C)[C@@H](OC)[C@@H]1C BKZOUCVNTCLNFF-IGXZVFLKSA-N 0.000 description 2
- SGKRLCUYIXIAHR-AKNGSSGZSA-N (4s,4ar,5s,5ar,6r,12ar)-4-(dimethylamino)-1,5,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4a,5,5a,6-tetrahydro-4h-tetracene-2-carboxamide Chemical compound C1=CC=C2[C@H](C)[C@@H]([C@H](O)[C@@H]3[C@](C(O)=C(C(N)=O)C(=O)[C@H]3N(C)C)(O)C3=O)C3=C(O)C2=C1O SGKRLCUYIXIAHR-AKNGSSGZSA-N 0.000 description 2
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 2
- STQGQHZAVUOBTE-UHFFFAOYSA-N 7-Cyan-hept-2t-en-4,6-diinsaeure Natural products C1=2C(O)=C3C(=O)C=4C(OC)=CC=CC=4C(=O)C3=C(O)C=2CC(O)(C(C)=O)CC1OC1CC(N)C(O)C(C)O1 STQGQHZAVUOBTE-UHFFFAOYSA-N 0.000 description 2
- ZGXJTSGNIOSYLO-UHFFFAOYSA-N 88755TAZ87 Chemical compound NCC(=O)CCC(O)=O ZGXJTSGNIOSYLO-UHFFFAOYSA-N 0.000 description 2
- 239000012117 Alexa Fluor 700 Substances 0.000 description 2
- 102100022524 Alpha-1-antichymotrypsin Human genes 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 2
- 206010004146 Basal cell carcinoma Diseases 0.000 description 2
- 206010005003 Bladder cancer Diseases 0.000 description 2
- 102100038078 CD276 antigen Human genes 0.000 description 2
- 101710185679 CD276 antigen Proteins 0.000 description 2
- 108010029697 CD40 Ligand Proteins 0.000 description 2
- 102100032937 CD40 ligand Human genes 0.000 description 2
- 102100039510 Cancer/testis antigen 2 Human genes 0.000 description 2
- 102100031762 Cancer/testis antigen family 45 member A3 Human genes 0.000 description 2
- 241000282472 Canis lupus familiaris Species 0.000 description 2
- GAGWJHPBXLXJQN-UORFTKCHSA-N Capecitabine Chemical compound C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1[C@H]1[C@H](O)[C@H](O)[C@@H](C)O1 GAGWJHPBXLXJQN-UORFTKCHSA-N 0.000 description 2
- 108010022366 Carcinoembryonic Antigen Proteins 0.000 description 2
- 102100025475 Carcinoembryonic antigen-related cell adhesion molecule 5 Human genes 0.000 description 2
- 102100039361 Chondrosarcoma-associated gene 2/3 protein Human genes 0.000 description 2
- 102100034330 Chromaffin granule amine transporter Human genes 0.000 description 2
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 2
- UHDGCWIWMRVCDJ-CCXZUQQUSA-N Cytarabine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 UHDGCWIWMRVCDJ-CCXZUQQUSA-N 0.000 description 2
- 108010092160 Dactinomycin Proteins 0.000 description 2
- 102100025012 Dipeptidyl peptidase 4 Human genes 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 206010014733 Endometrial cancer Diseases 0.000 description 2
- 206010014759 Endometrial neoplasm Diseases 0.000 description 2
- 241000242711 Fasciola hepatica Species 0.000 description 2
- 241000282326 Felis catus Species 0.000 description 2
- 241000710781 Flaviviridae Species 0.000 description 2
- 102100040578 G antigen 7 Human genes 0.000 description 2
- 102100031351 Galectin-9 Human genes 0.000 description 2
- 101000609762 Gallus gallus Ovalbumin Proteins 0.000 description 2
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 2
- 101000930822 Giardia intestinalis Dipeptidyl-peptidase 4 Proteins 0.000 description 2
- 102100041003 Glutamate carboxypeptidase 2 Human genes 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- 101150031823 HSP70 gene Proteins 0.000 description 2
- 102100022191 Hemogen Human genes 0.000 description 2
- 102100034458 Hepatitis A virus cellular receptor 2 Human genes 0.000 description 2
- 208000017604 Hodgkin disease Diseases 0.000 description 2
- 208000021519 Hodgkin lymphoma Diseases 0.000 description 2
- 208000010747 Hodgkins lymphoma Diseases 0.000 description 2
- 101000678026 Homo sapiens Alpha-1-antichymotrypsin Proteins 0.000 description 2
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 2
- 101000889345 Homo sapiens Cancer/testis antigen 2 Proteins 0.000 description 2
- 101000940803 Homo sapiens Cancer/testis antigen family 45 member A3 Proteins 0.000 description 2
- 101000745414 Homo sapiens Chondrosarcoma-associated gene 2/3 protein Proteins 0.000 description 2
- 101000641221 Homo sapiens Chromaffin granule amine transporter Proteins 0.000 description 2
- 101000892862 Homo sapiens Glutamate carboxypeptidase 2 Proteins 0.000 description 2
- 101001045553 Homo sapiens Hemogen Proteins 0.000 description 2
- 101001005718 Homo sapiens Melanoma-associated antigen 2 Proteins 0.000 description 2
- 101001005724 Homo sapiens Melanoma-associated antigen 9 Proteins 0.000 description 2
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 description 2
- 101000597425 Homo sapiens Nuclear RNA export factor 2 Proteins 0.000 description 2
- 101001114051 Homo sapiens P antigen family member 5 Proteins 0.000 description 2
- 101001126085 Homo sapiens Piwi-like protein 1 Proteins 0.000 description 2
- 101000880770 Homo sapiens Protein SSX2 Proteins 0.000 description 2
- 101000880774 Homo sapiens Protein SSX4 Proteins 0.000 description 2
- 101000825253 Homo sapiens Sperm protein associated with the nucleus on the X chromosome A Proteins 0.000 description 2
- 101000825254 Homo sapiens Sperm protein associated with the nucleus on the X chromosome B1 Proteins 0.000 description 2
- 101000825249 Homo sapiens Sperm protein associated with the nucleus on the X chromosome D Proteins 0.000 description 2
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 description 2
- 101000679851 Homo sapiens Tumor necrosis factor receptor superfamily member 4 Proteins 0.000 description 2
- 101000851376 Homo sapiens Tumor necrosis factor receptor superfamily member 8 Proteins 0.000 description 2
- 101000666896 Homo sapiens V-type immunoglobulin domain-containing suppressor of T-cell activation Proteins 0.000 description 2
- 101000814511 Homo sapiens X antigen family member 2 Proteins 0.000 description 2
- 241000701044 Human gammaherpesvirus 4 Species 0.000 description 2
- 206010020751 Hypersensitivity Diseases 0.000 description 2
- 102100034980 ICOS ligand Human genes 0.000 description 2
- 101710093458 ICOS ligand Proteins 0.000 description 2
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 2
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 2
- 108700005091 Immunoglobulin Genes Proteins 0.000 description 2
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 description 2
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 description 2
- 102000008070 Interferon-gamma Human genes 0.000 description 2
- ZQISRDCJNBUVMM-UHFFFAOYSA-N L-Histidinol Natural products OCC(N)CC1=CN=CN1 ZQISRDCJNBUVMM-UHFFFAOYSA-N 0.000 description 2
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 2
- ZQISRDCJNBUVMM-YFKPBYRVSA-N L-histidinol Chemical compound OC[C@@H](N)CC1=CNC=N1 ZQISRDCJNBUVMM-YFKPBYRVSA-N 0.000 description 2
- 125000000174 L-prolyl group Chemical group [H]N1C([H])([H])C([H])([H])C([H])([H])[C@@]1([H])C(*)=O 0.000 description 2
- 239000002136 L01XE07 - Lapatinib Substances 0.000 description 2
- 102000017578 LAG3 Human genes 0.000 description 2
- BKZOUCVNTCLNFF-UHFFFAOYSA-N Lonomycin Natural products COC1C(C)C(C2(C)OC(CC2)C2(C)OC3(OC(C(C)C(OC)C3)C(C)C3C(C(OC)C(C)C(O)(C(C)C(O)=O)O3)C)CC2)OC1C1OC(C)(O)C(C)C(OC)C1C BKZOUCVNTCLNFF-UHFFFAOYSA-N 0.000 description 2
- 108091054437 MHC class I family Proteins 0.000 description 2
- 102000043129 MHC class I family Human genes 0.000 description 2
- 102100028389 Melanoma antigen recognized by T-cells 1 Human genes 0.000 description 2
- 102100025081 Melanoma-associated antigen 2 Human genes 0.000 description 2
- 102100025079 Melanoma-associated antigen 9 Human genes 0.000 description 2
- 102000012220 Member 14 Tumor Necrosis Factor Receptors Human genes 0.000 description 2
- 108010061593 Member 14 Tumor Necrosis Factor Receptors Proteins 0.000 description 2
- 108090000157 Metallothionein Proteins 0.000 description 2
- 108010006519 Molecular Chaperones Proteins 0.000 description 2
- 241000713333 Mouse mammary tumor virus Species 0.000 description 2
- 101000597780 Mus musculus Tumor necrosis factor ligand superfamily member 18 Proteins 0.000 description 2
- 101000764258 Mus musculus Tumor necrosis factor ligand superfamily member 4 Proteins 0.000 description 2
- 241000186359 Mycobacterium Species 0.000 description 2
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 description 2
- 102100035403 Nuclear RNA export factor 2 Human genes 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 108010058846 Ovalbumin Proteins 0.000 description 2
- 206010033128 Ovarian cancer Diseases 0.000 description 2
- 206010061535 Ovarian neoplasm Diseases 0.000 description 2
- 102100023238 P antigen family member 5 Human genes 0.000 description 2
- 102000003992 Peroxidases Human genes 0.000 description 2
- 102100029364 Piwi-like protein 1 Human genes 0.000 description 2
- 206010035226 Plasma cell myeloma Diseases 0.000 description 2
- 108010072866 Prostate-Specific Antigen Proteins 0.000 description 2
- 102100038358 Prostate-specific antigen Human genes 0.000 description 2
- 102100037686 Protein SSX2 Human genes 0.000 description 2
- 102100037727 Protein SSX4 Human genes 0.000 description 2
- LOUPRKONTZGTKE-WZBLMQSHSA-N Quinine Chemical compound C([C@H]([C@H](C1)C=C)C2)C[N@@]1[C@@H]2[C@H](O)C1=CC=NC2=CC=C(OC)C=C21 LOUPRKONTZGTKE-WZBLMQSHSA-N 0.000 description 2
- 238000003559 RNA-seq method Methods 0.000 description 2
- 208000015634 Rectal Neoplasms Diseases 0.000 description 2
- 206010038389 Renal cancer Diseases 0.000 description 2
- 102100022327 Sperm protein associated with the nucleus on the X chromosome A Human genes 0.000 description 2
- 102100022326 Sperm protein associated with the nucleus on the X chromosome B1 Human genes 0.000 description 2
- 102100022325 Sperm protein associated with the nucleus on the X chromosome D Human genes 0.000 description 2
- 208000005718 Stomach Neoplasms Diseases 0.000 description 2
- 108010090804 Streptavidin Proteins 0.000 description 2
- 102100036234 Synaptonemal complex protein 1 Human genes 0.000 description 2
- 230000006044 T cell activation Effects 0.000 description 2
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 description 2
- FOCVUCIESVLUNU-UHFFFAOYSA-N Thiotepa Chemical compound C1CN1P(N1CC1)(=S)N1CC1 FOCVUCIESVLUNU-UHFFFAOYSA-N 0.000 description 2
- 102100023935 Transmembrane glycoprotein NMB Human genes 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- 102100040247 Tumor necrosis factor Human genes 0.000 description 2
- 102100035283 Tumor necrosis factor ligand superfamily member 18 Human genes 0.000 description 2
- 102100036857 Tumor necrosis factor receptor superfamily member 8 Human genes 0.000 description 2
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 2
- 108010079206 V-Set Domain-Containing T-Cell Activation Inhibitor 1 Proteins 0.000 description 2
- 102100038929 V-set domain-containing T-cell activation inhibitor 1 Human genes 0.000 description 2
- 102100038282 V-type immunoglobulin domain-containing suppressor of T-cell activation Human genes 0.000 description 2
- 102100039492 X antigen family member 2 Human genes 0.000 description 2
- 230000035508 accumulation Effects 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- RJURFGZVJUQBHK-UHFFFAOYSA-N actinomycin D Natural products CC1OC(=O)C(C(C)C)N(C)C(=O)CN(C)C(=O)C2CCCN2C(=O)C(C(C)C)NC(=O)C1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)NC4C(=O)NC(C(N5CCCC5C(=O)N(C)CC(=O)N(C)C(C(C)C)C(=O)OC4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-UHFFFAOYSA-N 0.000 description 2
- 238000007792 addition Methods 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000003042 antagnostic effect Effects 0.000 description 2
- 230000001093 anti-cancer Effects 0.000 description 2
- 230000000340 anti-metabolite Effects 0.000 description 2
- 230000014102 antigen processing and presentation of exogenous peptide antigen via MHC class I Effects 0.000 description 2
- 229960005475 antiinfective agent Drugs 0.000 description 2
- 229940100197 antimetabolite Drugs 0.000 description 2
- 239000002256 antimetabolite Substances 0.000 description 2
- 238000011203 antimicrobial therapy Methods 0.000 description 2
- 229950002916 avelumab Drugs 0.000 description 2
- WZPBZJONDBGPKJ-VEHQQRBSSA-N aztreonam Chemical compound O=C1N(S([O-])(=O)=O)[C@@H](C)[C@@H]1NC(=O)C(=N/OC(C)(C)C(O)=O)\C1=CSC([NH3+])=N1 WZPBZJONDBGPKJ-VEHQQRBSSA-N 0.000 description 2
- 229960003644 aztreonam Drugs 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- UCMIRNVEIXFBKS-UHFFFAOYSA-N beta-alanine Chemical compound NCCC(O)=O UCMIRNVEIXFBKS-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 239000008004 cell lysis buffer Substances 0.000 description 2
- 230000007969 cellular immunity Effects 0.000 description 2
- 201000007455 central nervous system cancer Diseases 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 238000010968 computed tomography angiography Methods 0.000 description 2
- 230000008602 contraction Effects 0.000 description 2
- XVOYSCVBGLVSOL-UHFFFAOYSA-N cysteic acid Chemical compound OC(=O)C(N)CS(O)(=O)=O XVOYSCVBGLVSOL-UHFFFAOYSA-N 0.000 description 2
- 206010052015 cytokine release syndrome Diseases 0.000 description 2
- 229940127089 cytotoxic agent Drugs 0.000 description 2
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 description 2
- 230000007123 defense Effects 0.000 description 2
- WHBIGIKBNXZKFE-UHFFFAOYSA-N delavirdine Chemical compound CC(C)NC1=CC=CN=C1N1CCN(C(=O)C=2NC3=CC=C(NS(C)(=O)=O)C=C3C=2)CC1 WHBIGIKBNXZKFE-UHFFFAOYSA-N 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- NOPFSRXAKWQILS-UHFFFAOYSA-N docosan-1-ol Chemical compound CCCCCCCCCCCCCCCCCCCCCCO NOPFSRXAKWQILS-UHFFFAOYSA-N 0.000 description 2
- 229960003722 doxycycline Drugs 0.000 description 2
- 229940126534 drug product Drugs 0.000 description 2
- 229950009791 durvalumab Drugs 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 230000007717 exclusion Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 2
- CHPZKNULDCNCBW-UHFFFAOYSA-N gallium nitrate Chemical compound [Ga+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O CHPZKNULDCNCBW-UHFFFAOYSA-N 0.000 description 2
- 229960003692 gamma aminobutyric acid Drugs 0.000 description 2
- 206010017758 gastric cancer Diseases 0.000 description 2
- SDUQYLNIPVEERB-QPPQHZFASA-N gemcitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1C(F)(F)[C@H](O)[C@@H](CO)O1 SDUQYLNIPVEERB-QPPQHZFASA-N 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 102000054766 genetic haplotypes Human genes 0.000 description 2
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 2
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 2
- 102000051450 human TNFSF4 Human genes 0.000 description 2
- 125000001165 hydrophobic group Chemical group 0.000 description 2
- 229960001101 ifosfamide Drugs 0.000 description 2
- HOMGKSMUEGBAAB-UHFFFAOYSA-N ifosfamide Chemical compound ClCCNP1(=O)OCCCN1CCCl HOMGKSMUEGBAAB-UHFFFAOYSA-N 0.000 description 2
- 210000002865 immune cell Anatomy 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 238000001114 immunoprecipitation Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 108091008042 inhibitory receptors Proteins 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 229960003130 interferon gamma Drugs 0.000 description 2
- PGHMRUGBZOYCAA-UHFFFAOYSA-N ionomycin Natural products O1C(CC(O)C(C)C(O)C(C)C=CCC(C)CC(C)C(O)=CC(=O)C(C)CC(C)CC(CCC(O)=O)C)CCC1(C)C1OC(C)(C(C)O)CC1 PGHMRUGBZOYCAA-UHFFFAOYSA-N 0.000 description 2
- PGHMRUGBZOYCAA-ADZNBVRBSA-N ionomycin Chemical compound O1[C@H](C[C@H](O)[C@H](C)[C@H](O)[C@H](C)/C=C/C[C@@H](C)C[C@@H](C)C(/O)=C/C(=O)[C@@H](C)C[C@@H](C)C[C@@H](CCC(O)=O)C)CC[C@@]1(C)[C@@H]1O[C@](C)([C@@H](C)O)CC1 PGHMRUGBZOYCAA-ADZNBVRBSA-N 0.000 description 2
- 229960004768 irinotecan Drugs 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 201000010982 kidney cancer Diseases 0.000 description 2
- BCFGMOOMADDAQU-UHFFFAOYSA-N lapatinib Chemical compound O1C(CNCCS(=O)(=O)C)=CC=C1C1=CC=C(N=CN=C2NC=3C=C(Cl)C(OCC=4C=C(F)C=CC=4)=CC=3)C2=C1 BCFGMOOMADDAQU-UHFFFAOYSA-N 0.000 description 2
- 208000032839 leukemia Diseases 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 201000007270 liver cancer Diseases 0.000 description 2
- 208000014018 liver neoplasm Diseases 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 230000001926 lymphatic effect Effects 0.000 description 2
- 230000000527 lymphocytic effect Effects 0.000 description 2
- 239000012139 lysis buffer Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 206010027191 meningioma Diseases 0.000 description 2
- 229960001428 mercaptopurine Drugs 0.000 description 2
- HPNSFSBZBAHARI-UHFFFAOYSA-N micophenolic acid Natural products OC1=C(CC=C(C)CCC(O)=O)C(OC)=C(C)C2=C1C(=O)OC2 HPNSFSBZBAHARI-UHFFFAOYSA-N 0.000 description 2
- 229960001156 mitoxantrone Drugs 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 229940035032 monophosphoryl lipid a Drugs 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 229960000951 mycophenolic acid Drugs 0.000 description 2
- HPNSFSBZBAHARI-RUDMXATFSA-N mycophenolic acid Chemical compound OC1=C(C\C=C(/C)CCC(O)=O)C(OC)=C(C)C2=C1C(=O)OC2 HPNSFSBZBAHARI-RUDMXATFSA-N 0.000 description 2
- 201000000050 myeloid neoplasm Diseases 0.000 description 2
- QZGIWPZCWHMVQL-UIYAJPBUSA-N neocarzinostatin chromophore Chemical compound O1[C@H](C)[C@H](O)[C@H](O)[C@@H](NC)[C@H]1O[C@@H]1C/2=C/C#C[C@H]3O[C@@]3([C@@H]3OC(=O)OC3)C#CC\2=C[C@H]1OC(=O)C1=C(O)C=CC2=C(C)C=C(OC)C=C12 QZGIWPZCWHMVQL-UIYAJPBUSA-N 0.000 description 2
- 230000001613 neoplastic effect Effects 0.000 description 2
- 231100000706 no observed effect level Toxicity 0.000 description 2
- 238000001151 non-parametric statistical test Methods 0.000 description 2
- 229940092253 ovalbumin Drugs 0.000 description 2
- 238000004806 packaging method and process Methods 0.000 description 2
- 208000008443 pancreatic carcinoma Diseases 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 description 2
- 239000000825 pharmaceutical preparation Substances 0.000 description 2
- ZJAOAACCNHFJAH-UHFFFAOYSA-N phosphonoformic acid Chemical compound OC(=O)P(O)(O)=O ZJAOAACCNHFJAH-UHFFFAOYSA-N 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 229920002627 poly(phosphazenes) Polymers 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 108010026466 polyproline Proteins 0.000 description 2
- 208000017805 post-transplant lymphoproliferative disease Diseases 0.000 description 2
- 230000002516 postimmunization Effects 0.000 description 2
- 230000001323 posttranslational effect Effects 0.000 description 2
- 239000011736 potassium bicarbonate Substances 0.000 description 2
- 235000015497 potassium bicarbonate Nutrition 0.000 description 2
- 229910000028 potassium bicarbonate Inorganic materials 0.000 description 2
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 235000013930 proline Nutrition 0.000 description 2
- 230000000069 prophylactic effect Effects 0.000 description 2
- 238000011321 prophylaxis Methods 0.000 description 2
- 238000002331 protein detection Methods 0.000 description 2
- 244000000040 protozoan parasite Species 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 206010038038 rectal cancer Diseases 0.000 description 2
- 201000001275 rectum cancer Diseases 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 210000002345 respiratory system Anatomy 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 235000004400 serine Nutrition 0.000 description 2
- 125000003607 serino group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C(O[H])([H])[H] 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- 208000000587 small cell lung carcinoma Diseases 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 238000011301 standard therapy Methods 0.000 description 2
- 229940032147 starch Drugs 0.000 description 2
- 201000011549 stomach cancer Diseases 0.000 description 2
- PVYJZLYGTZKPJE-UHFFFAOYSA-N streptonigrin Chemical compound C=1C=C2C(=O)C(OC)=C(N)C(=O)C2=NC=1C(C=1N)=NC(C(O)=O)=C(C)C=1C1=CC=C(OC)C(OC)=C1O PVYJZLYGTZKPJE-UHFFFAOYSA-N 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 238000007910 systemic administration Methods 0.000 description 2
- 239000008399 tap water Substances 0.000 description 2
- 235000020679 tap water Nutrition 0.000 description 2
- 229960001196 thiotepa Drugs 0.000 description 2
- 229960003087 tioguanine Drugs 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 238000011830 transgenic mouse model Methods 0.000 description 2
- 230000032258 transport Effects 0.000 description 2
- 229950007217 tremelimumab Drugs 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- 201000005112 urinary bladder cancer Diseases 0.000 description 2
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- XEEQGYMUWCZPDN-DOMZBBRYSA-N (-)-(11S,2'R)-erythro-mefloquine Chemical compound C([C@@H]1[C@@H](O)C=2C3=CC=CC(=C3N=C(C=2)C(F)(F)F)C(F)(F)F)CCCN1 XEEQGYMUWCZPDN-DOMZBBRYSA-N 0.000 description 1
- NNJPGOLRFBJNIW-HNNXBMFYSA-N (-)-demecolcine Chemical compound C1=C(OC)C(=O)C=C2[C@@H](NC)CCC3=CC(OC)=C(OC)C(OC)=C3C2=C1 NNJPGOLRFBJNIW-HNNXBMFYSA-N 0.000 description 1
- DIGQNXIGRZPYDK-WKSCXVIASA-N (2R)-6-amino-2-[[2-[[(2S)-2-[[2-[[(2R)-2-[[(2S)-2-[[(2R,3S)-2-[[2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[(2R)-2-[[(2S,3S)-2-[[(2R)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-2-[[(2R)-2-[[2-[[2-[[2-[(2-amino-1-hydroxyethylidene)amino]-3-carboxy-1-hydroxypropylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxypropylidene]amino]-1,3-dihydroxypropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxybutylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1-hydroxypropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-1,5-dihydroxy-5-iminopentylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxybutylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1-hydroxyethylidene]amino]hexanoic acid Chemical compound C[C@@H]([C@@H](C(=N[C@@H](CS)C(=N[C@@H](C)C(=N[C@@H](CO)C(=NCC(=N[C@@H](CCC(=N)O)C(=NC(CS)C(=N[C@H]([C@H](C)O)C(=N[C@H](CS)C(=N[C@H](CO)C(=NCC(=N[C@H](CS)C(=NCC(=N[C@H](CCCCN)C(=O)O)O)O)O)O)O)O)O)O)O)O)O)O)O)N=C([C@H](CS)N=C([C@H](CO)N=C([C@H](CO)N=C([C@H](C)N=C(CN=C([C@H](CO)N=C([C@H](CS)N=C(CN=C(C(CS)N=C(C(CC(=O)O)N=C(CN)O)O)O)O)O)O)O)O)O)O)O)O DIGQNXIGRZPYDK-WKSCXVIASA-N 0.000 description 1
- FDKWRPBBCBCIGA-REOHCLBHSA-N (2r)-2-azaniumyl-3-$l^{1}-selanylpropanoate Chemical compound [Se]C[C@H](N)C(O)=O FDKWRPBBCBCIGA-REOHCLBHSA-N 0.000 description 1
- BVAUMRCGVHUWOZ-ZETCQYMHSA-N (2s)-2-(cyclohexylazaniumyl)propanoate Chemical compound OC(=O)[C@H](C)NC1CCCCC1 BVAUMRCGVHUWOZ-ZETCQYMHSA-N 0.000 description 1
- MRTPISKDZDHEQI-YFKPBYRVSA-N (2s)-2-(tert-butylamino)propanoic acid Chemical compound OC(=O)[C@H](C)NC(C)(C)C MRTPISKDZDHEQI-YFKPBYRVSA-N 0.000 description 1
- ICLAYQQKWJGHBV-XJZMHMBSSA-N (2s)-2-[[(3r)-3-decoxytetradecanoyl]amino]-3-[(2r,3r,4r,5s,6r)-3-[[(3r)-3-decoxytetradecanoyl]amino]-4-[(3r)-3-decoxytetradecanoyl]oxy-6-(hydroxymethyl)-5-phosphonooxyoxan-2-yl]oxypropanoic acid Chemical compound CCCCCCCCCCC[C@@H](OCCCCCCCCCC)CC(=O)N[C@H](C(O)=O)CO[C@@H]1O[C@H](CO)[C@@H](OP(O)(O)=O)[C@H](OC(=O)C[C@@H](CCCCCCCCCCC)OCCCCCCCCCC)[C@H]1NC(=O)C[C@@H](CCCCCCCCCCC)OCCCCCCCCCC ICLAYQQKWJGHBV-XJZMHMBSSA-N 0.000 description 1
- YXTKHLHCVFUPPT-YYFJYKOTSA-N (2s)-2-[[4-[(2-amino-5-formyl-4-oxo-1,6,7,8-tetrahydropteridin-6-yl)methylamino]benzoyl]amino]pentanedioic acid;(1r,2r)-1,2-dimethanidylcyclohexane;5-fluoro-1h-pyrimidine-2,4-dione;oxalic acid;platinum(2+) Chemical compound [Pt+2].OC(=O)C(O)=O.[CH2-][C@@H]1CCCC[C@H]1[CH2-].FC1=CNC(=O)NC1=O.C1NC=2NC(N)=NC(=O)C=2N(C=O)C1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 YXTKHLHCVFUPPT-YYFJYKOTSA-N 0.000 description 1
- FLWWDYNPWOSLEO-HQVZTVAUSA-N (2s)-2-[[4-[1-(2-amino-4-oxo-1h-pteridin-6-yl)ethyl-methylamino]benzoyl]amino]pentanedioic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1C(C)N(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FLWWDYNPWOSLEO-HQVZTVAUSA-N 0.000 description 1
- NPDBDJFLKKQMCM-SCSAIBSYSA-N (2s)-2-amino-3,3-dimethylbutanoic acid Chemical compound CC(C)(C)[C@H](N)C(O)=O NPDBDJFLKKQMCM-SCSAIBSYSA-N 0.000 description 1
- CGMTUJFWROPELF-YPAAEMCBSA-N (3E,5S)-5-[(2S)-butan-2-yl]-3-(1-hydroxyethylidene)pyrrolidine-2,4-dione Chemical compound CC[C@H](C)[C@@H]1NC(=O)\C(=C(/C)O)C1=O CGMTUJFWROPELF-YPAAEMCBSA-N 0.000 description 1
- TVIRNGFXQVMMGB-OFWIHYRESA-N (3s,6r,10r,13e,16s)-16-[(2r,3r,4s)-4-chloro-3-hydroxy-4-phenylbutan-2-yl]-10-[(3-chloro-4-methoxyphenyl)methyl]-6-methyl-3-(2-methylpropyl)-1,4-dioxa-8,11-diazacyclohexadec-13-ene-2,5,9,12-tetrone Chemical compound C1=C(Cl)C(OC)=CC=C1C[C@@H]1C(=O)NC[C@@H](C)C(=O)O[C@@H](CC(C)C)C(=O)O[C@H]([C@H](C)[C@@H](O)[C@@H](Cl)C=2C=CC=CC=2)C/C=C/C(=O)N1 TVIRNGFXQVMMGB-OFWIHYRESA-N 0.000 description 1
- FFTVPQUHLQBXQZ-KVUCHLLUSA-N (4s,4as,5ar,12ar)-4,7-bis(dimethylamino)-1,10,11,12a-tetrahydroxy-3,12-dioxo-4a,5,5a,6-tetrahydro-4h-tetracene-2-carboxamide Chemical compound C1C2=C(N(C)C)C=CC(O)=C2C(O)=C2[C@@H]1C[C@H]1[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]1(O)C2=O FFTVPQUHLQBXQZ-KVUCHLLUSA-N 0.000 description 1
- XRBSKUSTLXISAB-XVVDYKMHSA-N (5r,6r,7r,8r)-8-hydroxy-7-(hydroxymethyl)-5-(3,4,5-trimethoxyphenyl)-5,6,7,8-tetrahydrobenzo[f][1,3]benzodioxole-6-carboxylic acid Chemical compound COC1=C(OC)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@H](O)[C@@H](CO)[C@@H]2C(O)=O)=C1 XRBSKUSTLXISAB-XVVDYKMHSA-N 0.000 description 1
- YYGNTYWPHWGJRM-UHFFFAOYSA-N (6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyltetracosa-2,6,10,14,18,22-hexaene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC=C(C)CCC=C(C)CCC=C(C)C YYGNTYWPHWGJRM-UHFFFAOYSA-N 0.000 description 1
- WDLWHQDACQUCJR-ZAMMOSSLSA-N (6r,7r)-7-[[(2r)-2-azaniumyl-2-(4-hydroxyphenyl)acetyl]amino]-8-oxo-3-[(e)-prop-1-enyl]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@@H]3N(C2=O)C(=C(CS3)/C=C/C)C(O)=O)=CC=C(O)C=C1 WDLWHQDACQUCJR-ZAMMOSSLSA-N 0.000 description 1
- XRBSKUSTLXISAB-UHFFFAOYSA-N (7R,7'R,8R,8'R)-form-Podophyllic acid Natural products COC1=C(OC)C(OC)=CC(C2C3=CC=4OCOC=4C=C3C(O)C(CO)C2C(O)=O)=C1 XRBSKUSTLXISAB-UHFFFAOYSA-N 0.000 description 1
- AESVUZLWRXEGEX-DKCAWCKPSA-N (7S,9R)-7-[(2S,4R,5R,6R)-4-amino-5-hydroxy-6-methyloxan-2-yl]oxy-6,9,11-trihydroxy-9-(2-hydroxyacetyl)-4-methoxy-8,10-dihydro-7H-tetracene-5,12-dione iron(3+) Chemical compound [Fe+3].COc1cccc2C(=O)c3c(O)c4C[C@@](O)(C[C@H](O[C@@H]5C[C@@H](N)[C@@H](O)[C@@H](C)O5)c4c(O)c3C(=O)c12)C(=O)CO AESVUZLWRXEGEX-DKCAWCKPSA-N 0.000 description 1
- JXVAMODRWBNUSF-KZQKBALLSA-N (7s,9r,10r)-7-[(2r,4s,5s,6s)-5-[[(2s,4as,5as,7s,9s,9ar,10ar)-2,9-dimethyl-3-oxo-4,4a,5a,6,7,9,9a,10a-octahydrodipyrano[4,2-a:4',3'-e][1,4]dioxin-7-yl]oxy]-4-(dimethylamino)-6-methyloxan-2-yl]oxy-10-[(2s,4s,5s,6s)-4-(dimethylamino)-5-hydroxy-6-methyloxan-2 Chemical compound O([C@@H]1C2=C(O)C=3C(=O)C4=CC=CC(O)=C4C(=O)C=3C(O)=C2[C@@H](O[C@@H]2O[C@@H](C)[C@@H](O[C@@H]3O[C@@H](C)[C@H]4O[C@@H]5O[C@@H](C)C(=O)C[C@@H]5O[C@H]4C3)[C@H](C2)N(C)C)C[C@]1(O)CC)[C@H]1C[C@H](N(C)C)[C@H](O)[C@H](C)O1 JXVAMODRWBNUSF-KZQKBALLSA-N 0.000 description 1
- INAUWOVKEZHHDM-PEDBPRJASA-N (7s,9s)-6,9,11-trihydroxy-9-(2-hydroxyacetyl)-7-[(2r,4s,5s,6s)-5-hydroxy-6-methyl-4-morpholin-4-yloxan-2-yl]oxy-4-methoxy-8,10-dihydro-7h-tetracene-5,12-dione;hydrochloride Chemical compound Cl.N1([C@H]2C[C@@H](O[C@@H](C)[C@H]2O)O[C@H]2C[C@@](O)(CC=3C(O)=C4C(=O)C=5C=CC=C(C=5C(=O)C4=C(O)C=32)OC)C(=O)CO)CCOCC1 INAUWOVKEZHHDM-PEDBPRJASA-N 0.000 description 1
- RCFNNLSZHVHCEK-IMHLAKCZSA-N (7s,9s)-7-(4-amino-6-methyloxan-2-yl)oxy-6,9,11-trihydroxy-9-(2-hydroxyacetyl)-4-methoxy-8,10-dihydro-7h-tetracene-5,12-dione;hydrochloride Chemical compound [Cl-].O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)C1CC([NH3+])CC(C)O1 RCFNNLSZHVHCEK-IMHLAKCZSA-N 0.000 description 1
- NOPNWHSMQOXAEI-PUCKCBAPSA-N (7s,9s)-7-[(2r,4s,5s,6s)-4-(2,3-dihydropyrrol-1-yl)-5-hydroxy-6-methyloxan-2-yl]oxy-6,9,11-trihydroxy-9-(2-hydroxyacetyl)-4-methoxy-8,10-dihydro-7h-tetracene-5,12-dione Chemical compound N1([C@H]2C[C@@H](O[C@@H](C)[C@H]2O)O[C@H]2C[C@@](O)(CC=3C(O)=C4C(=O)C=5C=CC=C(C=5C(=O)C4=C(O)C=32)OC)C(=O)CO)CCC=C1 NOPNWHSMQOXAEI-PUCKCBAPSA-N 0.000 description 1
- FPVKHBSQESCIEP-UHFFFAOYSA-N (8S)-3-(2-deoxy-beta-D-erythro-pentofuranosyl)-3,6,7,8-tetrahydroimidazo[4,5-d][1,3]diazepin-8-ol Natural products C1C(O)C(CO)OC1N1C(NC=NCC2O)=C2N=C1 FPVKHBSQESCIEP-UHFFFAOYSA-N 0.000 description 1
- FDKXTQMXEQVLRF-ZHACJKMWSA-N (E)-dacarbazine Chemical compound CN(C)\N=N\c1[nH]cnc1C(N)=O FDKXTQMXEQVLRF-ZHACJKMWSA-N 0.000 description 1
- WHTVZRBIWZFKQO-AWEZNQCLSA-N (S)-chloroquine Chemical compound ClC1=CC=C2C(N[C@@H](C)CCCN(CC)CC)=CC=NC2=C1 WHTVZRBIWZFKQO-AWEZNQCLSA-N 0.000 description 1
- AGNGYMCLFWQVGX-AGFFZDDWSA-N (e)-1-[(2s)-2-amino-2-carboxyethoxy]-2-diazonioethenolate Chemical compound OC(=O)[C@@H](N)CO\C([O-])=C\[N+]#N AGNGYMCLFWQVGX-AGFFZDDWSA-N 0.000 description 1
- NCCJWSXETVVUHK-ZYSAIPPVSA-N (z)-7-[(2r)-2-amino-2-carboxyethyl]sulfanyl-2-[[(1s)-2,2-dimethylcyclopropanecarbonyl]amino]hept-2-enoic acid;(5r,6s)-3-[2-(aminomethylideneamino)ethylsulfanyl]-6-[(1r)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid Chemical compound C1C(SCC\N=C/N)=C(C(O)=O)N2C(=O)[C@H]([C@H](O)C)[C@H]21.CC1(C)C[C@@H]1C(=O)N\C(=C/CCCCSC[C@H](N)C(O)=O)C(O)=O NCCJWSXETVVUHK-ZYSAIPPVSA-N 0.000 description 1
- FONKWHRXTPJODV-DNQXCXABSA-N 1,3-bis[2-[(8s)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-3h-pyrrolo[3,2-e]indole-6-carbonyl]-1h-indol-5-yl]urea Chemical compound C1([C@H](CCl)CN2C(=O)C=3NC4=CC=C(C=C4C=3)NC(=O)NC=3C=C4C=C(NC4=CC=3)C(=O)N3C4=CC(O)=C5NC=C(C5=C4[C@H](CCl)C3)C)=C2C=C(O)C2=C1C(C)=CN2 FONKWHRXTPJODV-DNQXCXABSA-N 0.000 description 1
- WEYNBWVKOYCCQT-UHFFFAOYSA-N 1-(3-chloro-4-methylphenyl)-3-{2-[({5-[(dimethylamino)methyl]-2-furyl}methyl)thio]ethyl}urea Chemical compound O1C(CN(C)C)=CC=C1CSCCNC(=O)NC1=CC=C(C)C(Cl)=C1 WEYNBWVKOYCCQT-UHFFFAOYSA-N 0.000 description 1
- IOTAOYHKWICOBK-UHFFFAOYSA-N 1-[amino-(4-chloroanilino)methylidene]-2-propan-2-ylguanidine;3-[4-(4-chlorophenyl)cyclohexyl]-4-hydroxynaphthalene-1,2-dione;hydrochloride Chemical compound Cl.CC(C)N=C(N)\N=C(/N)NC1=CC=C(Cl)C=C1.O=C1C(=O)C2=CC=CC=C2C(O)=C1C(CC1)CCC1C1=CC=C(Cl)C=C1 IOTAOYHKWICOBK-UHFFFAOYSA-N 0.000 description 1
- BTOTXLJHDSNXMW-POYBYMJQSA-N 2,3-dideoxyuridine Chemical compound O1[C@H](CO)CC[C@@H]1N1C(=O)NC(=O)C=C1 BTOTXLJHDSNXMW-POYBYMJQSA-N 0.000 description 1
- OGNSCSPNOLGXSM-UHFFFAOYSA-N 2,4-diaminobutyric acid Chemical compound NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 description 1
- BOMZMNZEXMAQQW-UHFFFAOYSA-N 2,5,11-trimethyl-6h-pyrido[4,3-b]carbazol-2-ium-9-ol;acetate Chemical compound CC([O-])=O.C[N+]1=CC=C2C(C)=C(NC=3C4=CC(O)=CC=3)C4=C(C)C2=C1 BOMZMNZEXMAQQW-UHFFFAOYSA-N 0.000 description 1
- QCXJFISCRQIYID-IAEPZHFASA-N 2-amino-1-n-[(3s,6s,7r,10s,16s)-3-[(2s)-butan-2-yl]-7,11,14-trimethyl-2,5,9,12,15-pentaoxo-10-propan-2-yl-8-oxa-1,4,11,14-tetrazabicyclo[14.3.0]nonadecan-6-yl]-4,6-dimethyl-3-oxo-9-n-[(3s,6s,7r,10s,16s)-7,11,14-trimethyl-2,5,9,12,15-pentaoxo-3,10-di(propa Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N=C2C(C(=O)N[C@@H]3C(=O)N[C@H](C(N4CCC[C@H]4C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]3C)=O)[C@@H](C)CC)=C(N)C(=O)C(C)=C2O2)C2=C(C)C=C1 QCXJFISCRQIYID-IAEPZHFASA-N 0.000 description 1
- GOJUJUVQIVIZAV-UHFFFAOYSA-N 2-amino-4,6-dichloropyrimidine-5-carbaldehyde Chemical group NC1=NC(Cl)=C(C=O)C(Cl)=N1 GOJUJUVQIVIZAV-UHFFFAOYSA-N 0.000 description 1
- VNBAOSVONFJBKP-UHFFFAOYSA-N 2-chloro-n,n-bis(2-chloroethyl)propan-1-amine;hydrochloride Chemical compound Cl.CC(Cl)CN(CCCl)CCCl VNBAOSVONFJBKP-UHFFFAOYSA-N 0.000 description 1
- AZSNMRSAGSSBNP-UHFFFAOYSA-N 22,23-dihydroavermectin B1a Natural products C1CC(C)C(C(C)CC)OC21OC(CC=C(C)C(OC1OC(C)C(OC3OC(C)C(O)C(OC)C3)C(OC)C1)C(C)C=CC=C1C3(C(C(=O)O4)C=C(C)C(O)C3OC1)O)CC4C2 AZSNMRSAGSSBNP-UHFFFAOYSA-N 0.000 description 1
- YIMDLWDNDGKDTJ-QLKYHASDSA-N 3'-deamino-3'-(3-cyanomorpholin-4-yl)doxorubicin Chemical compound N1([C@H]2C[C@@H](O[C@@H](C)[C@H]2O)O[C@H]2C[C@@](O)(CC=3C(O)=C4C(=O)C=5C=CC=C(C=5C(=O)C4=C(O)C=32)OC)C(=O)CO)CCOCC1C#N YIMDLWDNDGKDTJ-QLKYHASDSA-N 0.000 description 1
- NDMPLJNOPCLANR-UHFFFAOYSA-N 3,4-dihydroxy-15-(4-hydroxy-18-methoxycarbonyl-5,18-seco-ibogamin-18-yl)-16-methoxy-1-methyl-6,7-didehydro-aspidospermidine-3-carboxylic acid methyl ester Natural products C1C(CC)(O)CC(CC2(C(=O)OC)C=3C(=CC4=C(C56C(C(C(O)C7(CC)C=CCN(C67)CC5)(O)C(=O)OC)N4C)C=3)OC)CN1CCC1=C2NC2=CC=CC=C12 NDMPLJNOPCLANR-UHFFFAOYSA-N 0.000 description 1
- PWMYMKOUNYTVQN-UHFFFAOYSA-N 3-(8,8-diethyl-2-aza-8-germaspiro[4.5]decan-2-yl)-n,n-dimethylpropan-1-amine Chemical compound C1C[Ge](CC)(CC)CCC11CN(CCCN(C)C)CC1 PWMYMKOUNYTVQN-UHFFFAOYSA-N 0.000 description 1
- 101800000504 3C-like protease Proteins 0.000 description 1
- AOJJSUZBOXZQNB-VTZDEGQISA-N 4'-epidoxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-VTZDEGQISA-N 0.000 description 1
- LUBUTTBEBGYNJN-UHFFFAOYSA-N 4-amino-n-(5,6-dimethoxypyrimidin-4-yl)benzenesulfonamide;5-(4-chlorophenyl)-6-ethylpyrimidine-2,4-diamine Chemical compound CCC1=NC(N)=NC(N)=C1C1=CC=C(Cl)C=C1.COC1=NC=NC(NS(=O)(=O)C=2C=CC(N)=CC=2)=C1OC LUBUTTBEBGYNJN-UHFFFAOYSA-N 0.000 description 1
- TVZGACDUOSZQKY-LBPRGKRZSA-N 4-aminofolic acid Chemical compound C1=NC2=NC(N)=NC(N)=C2N=C1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 TVZGACDUOSZQKY-LBPRGKRZSA-N 0.000 description 1
- 101710169336 5'-deoxyadenosine deaminase Proteins 0.000 description 1
- 102100022464 5'-nucleotidase Human genes 0.000 description 1
- LGZKGOGODCLQHG-CYBMUJFWSA-N 5-[(2r)-2-hydroxy-2-(3,4,5-trimethoxyphenyl)ethyl]-2-methoxyphenol Chemical compound C1=C(O)C(OC)=CC=C1C[C@@H](O)C1=CC(OC)=C(OC)C(OC)=C1 LGZKGOGODCLQHG-CYBMUJFWSA-N 0.000 description 1
- IDPUKCWIGUEADI-UHFFFAOYSA-N 5-[bis(2-chloroethyl)amino]uracil Chemical compound ClCCN(CCCl)C1=CNC(=O)NC1=O IDPUKCWIGUEADI-UHFFFAOYSA-N 0.000 description 1
- NMUSYJAQQFHJEW-KVTDHHQDSA-N 5-azacytidine Chemical compound O=C1N=C(N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 NMUSYJAQQFHJEW-KVTDHHQDSA-N 0.000 description 1
- GJOHLWZHWQUKAU-UHFFFAOYSA-N 5-azaniumylpentan-2-yl-(6-methoxyquinolin-8-yl)azanium;dihydrogen phosphate Chemical compound OP(O)(O)=O.OP(O)(O)=O.N1=CC=CC2=CC(OC)=CC(NC(C)CCCN)=C21 GJOHLWZHWQUKAU-UHFFFAOYSA-N 0.000 description 1
- SLXKOJJOQWFEFD-UHFFFAOYSA-N 6-aminohexanoic acid Chemical compound NCCCCCC(O)=O SLXKOJJOQWFEFD-UHFFFAOYSA-N 0.000 description 1
- WYXSYVWAUAUWLD-SHUUEZRQSA-N 6-azauridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=N1 WYXSYVWAUAUWLD-SHUUEZRQSA-N 0.000 description 1
- 229960005538 6-diazo-5-oxo-L-norleucine Drugs 0.000 description 1
- YCWQAMGASJSUIP-YFKPBYRVSA-N 6-diazo-5-oxo-L-norleucine Chemical compound OC(=O)[C@@H](N)CCC(=O)C=[N+]=[N-] YCWQAMGASJSUIP-YFKPBYRVSA-N 0.000 description 1
- SPBDXSGPUHCETR-JFUDTMANSA-N 8883yp2r6d Chemical compound O1[C@@H](C)[C@H](O)[C@@H](OC)C[C@@H]1O[C@@H]1[C@@H](OC)C[C@H](O[C@@H]2C(=C/C[C@@H]3C[C@@H](C[C@@]4(O[C@@H]([C@@H](C)CC4)C(C)C)O3)OC(=O)[C@@H]3C=C(C)[C@@H](O)[C@H]4OC\C([C@@]34O)=C/C=C/[C@@H]2C)/C)O[C@H]1C.C1C[C@H](C)[C@@H]([C@@H](C)CC)O[C@@]21O[C@H](C\C=C(C)\[C@@H](O[C@@H]1O[C@@H](C)[C@H](O[C@@H]3O[C@@H](C)[C@H](O)[C@@H](OC)C3)[C@@H](OC)C1)[C@@H](C)\C=C\C=C/1[C@]3([C@H](C(=O)O4)C=C(C)[C@@H](O)[C@H]3OC\1)O)C[C@H]4C2 SPBDXSGPUHCETR-JFUDTMANSA-N 0.000 description 1
- GSDSWSVVBLHKDQ-UHFFFAOYSA-N 9-fluoro-3-methyl-10-(4-methylpiperazin-1-yl)-7-oxo-2,3-dihydro-7H-[1,4]oxazino[2,3,4-ij]quinoline-6-carboxylic acid Chemical compound FC1=CC(C(C(C(O)=O)=C2)=O)=C3N2C(C)COC3=C1N1CCN(C)CC1 GSDSWSVVBLHKDQ-UHFFFAOYSA-N 0.000 description 1
- HDZZVAMISRMYHH-UHFFFAOYSA-N 9beta-Ribofuranosyl-7-deazaadenin Natural products C1=CC=2C(N)=NC=NC=2N1C1OC(CO)C(O)C1O HDZZVAMISRMYHH-UHFFFAOYSA-N 0.000 description 1
- 102100031910 A-kinase anchor protein 3 Human genes 0.000 description 1
- 102100040079 A-kinase anchor protein 4 Human genes 0.000 description 1
- 208000002008 AIDS-Related Lymphoma Diseases 0.000 description 1
- 102100031315 AP-2 complex subunit mu Human genes 0.000 description 1
- 102100039864 ATPase family AAA domain-containing protein 2 Human genes 0.000 description 1
- 241000589291 Acinetobacter Species 0.000 description 1
- 241000588626 Acinetobacter baumannii Species 0.000 description 1
- 241000588624 Acinetobacter calcoaceticus Species 0.000 description 1
- 241001148231 Acinetobacter haemolyticus Species 0.000 description 1
- 102100022907 Acrosin-binding protein Human genes 0.000 description 1
- 102100022498 Actin-like protein 8 Human genes 0.000 description 1
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 description 1
- 102100029457 Adenine phosphoribosyltransferase Human genes 0.000 description 1
- 108010024223 Adenine phosphoribosyltransferase Proteins 0.000 description 1
- 101710137115 Adenylyl cyclase-associated protein 1 Proteins 0.000 description 1
- 102100021879 Adenylyl cyclase-associated protein 2 Human genes 0.000 description 1
- 101710137132 Adenylyl cyclase-associated protein 2 Proteins 0.000 description 1
- 241000607528 Aeromonas hydrophila Species 0.000 description 1
- 108010012934 Albumin-Bound Paclitaxel Proteins 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 239000012114 Alexa Fluor 647 Substances 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- CEIZFXOZIQNICU-UHFFFAOYSA-N Alternaria alternata Crofton-weed toxin Natural products CCC(C)C1NC(=O)C(C(C)=O)=C1O CEIZFXOZIQNICU-UHFFFAOYSA-N 0.000 description 1
- 241000498253 Ancylostoma duodenale Species 0.000 description 1
- 102100033330 Ankyrin repeat domain-containing protein 45 Human genes 0.000 description 1
- WZPBZJONDBGPKJ-UHFFFAOYSA-N Antibiotic SQ 26917 Natural products O=C1N(S(O)(=O)=O)C(C)C1NC(=O)C(=NOC(C)(C)C(O)=O)C1=CSC(N)=N1 WZPBZJONDBGPKJ-UHFFFAOYSA-N 0.000 description 1
- 101100490566 Arabidopsis thaliana ADR2 gene Proteins 0.000 description 1
- 101100504181 Arabidopsis thaliana GCS1 gene Proteins 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 102100023189 Armadillo repeat-containing protein 3 Human genes 0.000 description 1
- 241000244185 Ascaris lumbricoides Species 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 206010003571 Astrocytoma Diseases 0.000 description 1
- AXRYRYVKAWYZBR-UHFFFAOYSA-N Atazanavir Natural products C=1C=C(C=2N=CC=CC=2)C=CC=1CN(NC(=O)C(NC(=O)OC)C(C)(C)C)CC(O)C(NC(=O)C(NC(=O)OC)C(C)(C)C)CC1=CC=CC=C1 AXRYRYVKAWYZBR-UHFFFAOYSA-N 0.000 description 1
- 108010019625 Atazanavir Sulfate Proteins 0.000 description 1
- 102100022717 Atypical chemokine receptor 1 Human genes 0.000 description 1
- 241000972773 Aulopiformes Species 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- NOWKCMXCCJGMRR-UHFFFAOYSA-N Aziridine Chemical class C1CN1 NOWKCMXCCJGMRR-UHFFFAOYSA-N 0.000 description 1
- 102100035526 B melanoma antigen 1 Human genes 0.000 description 1
- 102100035565 B melanoma antigen 2 Human genes 0.000 description 1
- 102100035527 B melanoma antigen 3 Human genes 0.000 description 1
- 102100035567 B melanoma antigen 4 Human genes 0.000 description 1
- 102100035566 B melanoma antigen 5 Human genes 0.000 description 1
- 102100029822 B- and T-lymphocyte attenuator Human genes 0.000 description 1
- 208000010839 B-cell chronic lymphocytic leukemia Diseases 0.000 description 1
- 208000003950 B-cell lymphoma Diseases 0.000 description 1
- 210000002237 B-cell of pancreatic islet Anatomy 0.000 description 1
- 102100038080 B-cell receptor CD22 Human genes 0.000 description 1
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 description 1
- 241001135322 Bacteroides eggerthii Species 0.000 description 1
- 241000606124 Bacteroides fragilis Species 0.000 description 1
- 241000606123 Bacteroides thetaiotaomicron Species 0.000 description 1
- 241000606219 Bacteroides uniformis Species 0.000 description 1
- 241000606215 Bacteroides vulgatus Species 0.000 description 1
- VGGGPCQERPFHOB-MCIONIFRSA-N Bestatin Chemical compound CC(C)C[C@H](C(O)=O)NC(=O)[C@@H](O)[C@H](N)CC1=CC=CC=C1 VGGGPCQERPFHOB-MCIONIFRSA-N 0.000 description 1
- 229940122361 Bisphosphonate Drugs 0.000 description 1
- 108010006654 Bleomycin Proteins 0.000 description 1
- 206010005949 Bone cancer Diseases 0.000 description 1
- 208000018084 Bone neoplasm Diseases 0.000 description 1
- 241000588779 Bordetella bronchiseptica Species 0.000 description 1
- 241000588780 Bordetella parapertussis Species 0.000 description 1
- 241000588832 Bordetella pertussis Species 0.000 description 1
- 241000589969 Borreliella burgdorferi Species 0.000 description 1
- 208000003174 Brain Neoplasms Diseases 0.000 description 1
- 206010055113 Breast cancer metastatic Diseases 0.000 description 1
- 102100029894 Bromodomain testis-specific protein Human genes 0.000 description 1
- 241000589562 Brucella Species 0.000 description 1
- MBABCNBNDNGODA-LTGLSHGVSA-N Bullatacin Natural products O=C1C(C[C@H](O)CCCCCCCCCC[C@@H](O)[C@@H]2O[C@@H]([C@@H]3O[C@H]([C@@H](O)CCCCCCCCCC)CC3)CC2)=C[C@H](C)O1 MBABCNBNDNGODA-LTGLSHGVSA-N 0.000 description 1
- KGGVWMAPBXIMEM-JQFCFGFHSA-N Bullatacinone Natural products O=C(C[C@H]1C(=O)O[C@H](CCCCCCCCCC[C@H](O)[C@@H]2O[C@@H]([C@@H]3O[C@@H]([C@@H](O)CCCCCCCCCC)CC3)CC2)C1)C KGGVWMAPBXIMEM-JQFCFGFHSA-N 0.000 description 1
- KGGVWMAPBXIMEM-ZRTAFWODSA-N Bullatacinone Chemical compound O1[C@@H]([C@@H](O)CCCCCCCCCC)CC[C@@H]1[C@@H]1O[C@@H]([C@H](O)CCCCCCCCCC[C@H]2OC(=O)[C@H](CC(C)=O)C2)CC1 KGGVWMAPBXIMEM-ZRTAFWODSA-N 0.000 description 1
- 241000589513 Burkholderia cepacia Species 0.000 description 1
- COVZYZSDYWQREU-UHFFFAOYSA-N Busulfan Chemical compound CS(=O)(=O)OCCCCOS(C)(=O)=O COVZYZSDYWQREU-UHFFFAOYSA-N 0.000 description 1
- 102100025905 C-Jun-amino-terminal kinase-interacting protein 4 Human genes 0.000 description 1
- 108010074051 C-Reactive Protein Proteins 0.000 description 1
- 102100032752 C-reactive protein Human genes 0.000 description 1
- 102100032982 CCR4-NOT transcription complex subunit 9 Human genes 0.000 description 1
- 101150050673 CHK1 gene Proteins 0.000 description 1
- 102100039305 CPX chromosomal region candidate gene 1 protein Human genes 0.000 description 1
- 102000000905 Cadherin Human genes 0.000 description 1
- 108050007957 Cadherin Proteins 0.000 description 1
- 101100124795 Caenorhabditis elegans hsp-110 gene Proteins 0.000 description 1
- 102100025338 Calcium-binding tyrosine phosphorylation-regulated protein Human genes 0.000 description 1
- 102100038613 Calreticulin-3 Human genes 0.000 description 1
- KLWPJMFMVPTNCC-UHFFFAOYSA-N Camptothecin Natural products CCC1(O)C(=O)OCC2=C1C=C3C4Nc5ccccc5C=C4CN3C2=O KLWPJMFMVPTNCC-UHFFFAOYSA-N 0.000 description 1
- 241000589876 Campylobacter Species 0.000 description 1
- 241000589877 Campylobacter coli Species 0.000 description 1
- 241000589874 Campylobacter fetus Species 0.000 description 1
- 241000589875 Campylobacter jejuni Species 0.000 description 1
- 102100025933 Cancer-associated gene 1 protein Human genes 0.000 description 1
- 102100039634 Cancer/testis antigen 47B Human genes 0.000 description 1
- 102100031757 Cancer/testis antigen family 45 member A1 Human genes 0.000 description 1
- 102100031761 Cancer/testis antigen family 45 member A2 Human genes 0.000 description 1
- 102100031661 Cancer/testis antigen family 45 member A5 Human genes 0.000 description 1
- 102100031662 Cancer/testis antigen family 45 member A6 Human genes 0.000 description 1
- 241000282465 Canis Species 0.000 description 1
- GAGWJHPBXLXJQN-UHFFFAOYSA-N Capecitabine Natural products C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1C1C(O)C(O)C(C)O1 GAGWJHPBXLXJQN-UHFFFAOYSA-N 0.000 description 1
- SHHKQEUPHAENFK-UHFFFAOYSA-N Carboquone Chemical compound O=C1C(C)=C(N2CC2)C(=O)C(C(COC(N)=O)OC)=C1N1CC1 SHHKQEUPHAENFK-UHFFFAOYSA-N 0.000 description 1
- 208000009458 Carcinoma in Situ Diseases 0.000 description 1
- 208000020446 Cardiac disease Diseases 0.000 description 1
- 206010007559 Cardiac failure congestive Diseases 0.000 description 1
- AOCCBINRVIKJHY-UHFFFAOYSA-N Carmofur Chemical compound CCCCCCNC(=O)N1C=C(F)C(=O)NC1=O AOCCBINRVIKJHY-UHFFFAOYSA-N 0.000 description 1
- DLGOEMSEDOSKAD-UHFFFAOYSA-N Carmustine Chemical compound ClCCNC(=O)N(N=O)CCCl DLGOEMSEDOSKAD-UHFFFAOYSA-N 0.000 description 1
- 102100028002 Catenin alpha-2 Human genes 0.000 description 1
- 102100028906 Catenin delta-1 Human genes 0.000 description 1
- 108010067316 Catenins Proteins 0.000 description 1
- 102000016362 Catenins Human genes 0.000 description 1
- 241000700199 Cavia porcellus Species 0.000 description 1
- GNWUOVJNSFPWDD-XMZRARIVSA-M Cefoxitin sodium Chemical compound [Na+].N([C@]1(OC)C(N2C(=C(COC(N)=O)CS[C@@H]21)C([O-])=O)=O)C(=O)CC1=CC=CS1 GNWUOVJNSFPWDD-XMZRARIVSA-M 0.000 description 1
- 108010039939 Cell Wall Skeleton Proteins 0.000 description 1
- 102100025064 Cellular tumor antigen p53 Human genes 0.000 description 1
- 102100035673 Centrosomal protein of 290 kDa Human genes 0.000 description 1
- 101710198317 Centrosomal protein of 290 kDa Proteins 0.000 description 1
- 102100031219 Centrosomal protein of 55 kDa Human genes 0.000 description 1
- 101710092479 Centrosomal protein of 55 kDa Proteins 0.000 description 1
- 229930186147 Cephalosporin Natural products 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 108010058432 Chaperonin 60 Proteins 0.000 description 1
- 102000006303 Chaperonin 60 Human genes 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- JWBOIMRXGHLCPP-UHFFFAOYSA-N Chloditan Chemical compound C=1C=CC=C(Cl)C=1C(C(Cl)Cl)C1=CC=C(Cl)C=C1 JWBOIMRXGHLCPP-UHFFFAOYSA-N 0.000 description 1
- XCDXSSFOJZZGQC-UHFFFAOYSA-N Chlornaphazine Chemical compound C1=CC=CC2=CC(N(CCCl)CCCl)=CC=C21 XCDXSSFOJZZGQC-UHFFFAOYSA-N 0.000 description 1
- 241000282552 Chlorocebus aethiops Species 0.000 description 1
- MKQWTWSXVILIKJ-LXGUWJNJSA-N Chlorozotocin Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](C=O)NC(=O)N(N=O)CCCl MKQWTWSXVILIKJ-LXGUWJNJSA-N 0.000 description 1
- 102000009016 Cholera Toxin Human genes 0.000 description 1
- 108010049048 Cholera Toxin Proteins 0.000 description 1
- 208000006332 Choriocarcinoma Diseases 0.000 description 1
- 241000253343 Chromadorea Species 0.000 description 1
- VWFCHDSQECPREK-LURJTMIESA-N Cidofovir Chemical compound NC=1C=CN(C[C@@H](CO)OCP(O)(O)=O)C(=O)N=1 VWFCHDSQECPREK-LURJTMIESA-N 0.000 description 1
- 235000001258 Cinchona calisaya Nutrition 0.000 description 1
- 102100040901 Circadian clock protein PASD1 Human genes 0.000 description 1
- 208000005443 Circulating Neoplastic Cells Diseases 0.000 description 1
- 241000588923 Citrobacter Species 0.000 description 1
- 241000588919 Citrobacter freundii Species 0.000 description 1
- 241000193163 Clostridioides difficile Species 0.000 description 1
- 241000193403 Clostridium Species 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 102100032368 Coiled-coil domain-containing protein 110 Human genes 0.000 description 1
- 102100021967 Coiled-coil domain-containing protein 33 Human genes 0.000 description 1
- 102100035180 Coiled-coil domain-containing protein 62 Human genes 0.000 description 1
- 102100025844 Coiled-coil domain-containing protein 83 Human genes 0.000 description 1
- 206010055114 Colon cancer metastatic Diseases 0.000 description 1
- 241000186227 Corynebacterium diphtheriae Species 0.000 description 1
- 241000918600 Corynebacterium ulcerans Species 0.000 description 1
- 241000709687 Coxsackievirus Species 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- 229930188224 Cryptophycin Natural products 0.000 description 1
- 241000223938 Cryptosporidium muris Species 0.000 description 1
- 102100025571 Cutaneous T-cell lymphoma-associated antigen 1 Human genes 0.000 description 1
- 108050006400 Cyclin Proteins 0.000 description 1
- 102000016736 Cyclin Human genes 0.000 description 1
- 108010025464 Cyclin-Dependent Kinase 4 Proteins 0.000 description 1
- 102100036252 Cyclin-dependent kinase 4 Human genes 0.000 description 1
- 102100027350 Cysteine-rich secretory protein 2 Human genes 0.000 description 1
- 201000003808 Cystic echinococcosis Diseases 0.000 description 1
- 102100031654 Cytochrome c oxidase subunit 6B2 Human genes 0.000 description 1
- FDKWRPBBCBCIGA-UWTATZPHSA-N D-Selenocysteine Natural products [Se]C[C@@H](N)C(O)=O FDKWRPBBCBCIGA-UWTATZPHSA-N 0.000 description 1
- 102100039771 DDB1- and CUL4-associated factor 12 Human genes 0.000 description 1
- YVGGHNCTFXOJCH-UHFFFAOYSA-N DDT Chemical compound C1=CC(Cl)=CC=C1C(C(Cl)(Cl)Cl)C1=CC=C(Cl)C=C1 YVGGHNCTFXOJCH-UHFFFAOYSA-N 0.000 description 1
- 101150074155 DHFR gene Proteins 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- 239000012623 DNA damaging agent Substances 0.000 description 1
- 241000252212 Danio rerio Species 0.000 description 1
- 101000923091 Danio rerio Aristaless-related homeobox protein Proteins 0.000 description 1
- 101100016370 Danio rerio hsp90a.1 gene Proteins 0.000 description 1
- WEAHRLBPCANXCN-UHFFFAOYSA-N Daunomycin Natural products CCC1(O)CC(OC2CC(N)C(O)C(C)O2)c3cc4C(=O)c5c(OC)cccc5C(=O)c4c(O)c3C1 WEAHRLBPCANXCN-UHFFFAOYSA-N 0.000 description 1
- 241001600125 Delftia acidovorans Species 0.000 description 1
- NNJPGOLRFBJNIW-UHFFFAOYSA-N Demecolcine Natural products C1=C(OC)C(=O)C=C2C(NC)CCC3=CC(OC)=C(OC)C(OC)=C3C2=C1 NNJPGOLRFBJNIW-UHFFFAOYSA-N 0.000 description 1
- 241000725619 Dengue virus Species 0.000 description 1
- 241000702421 Dependoparvovirus Species 0.000 description 1
- 108010002156 Depsipeptides Proteins 0.000 description 1
- 102100036949 Developmental pluripotency-associated protein 2 Human genes 0.000 description 1
- AUGQEEXBDZWUJY-ZLJUKNTDSA-N Diacetoxyscirpenol Chemical compound C([C@]12[C@]3(C)[C@H](OC(C)=O)[C@@H](O)[C@H]1O[C@@H]1C=C(C)CC[C@@]13COC(=O)C)O2 AUGQEEXBDZWUJY-ZLJUKNTDSA-N 0.000 description 1
- AUGQEEXBDZWUJY-UHFFFAOYSA-N Diacetoxyscirpenol Natural products CC(=O)OCC12CCC(C)=CC1OC1C(O)C(OC(C)=O)C2(C)C11CO1 AUGQEEXBDZWUJY-UHFFFAOYSA-N 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 102100037981 Dickkopf-like protein 1 Human genes 0.000 description 1
- 101100216227 Dictyostelium discoideum anapc3 gene Proteins 0.000 description 1
- 101100285708 Dictyostelium discoideum hspD gene Proteins 0.000 description 1
- 101100125027 Dictyostelium discoideum mhsp70 gene Proteins 0.000 description 1
- BXZVVICBKDXVGW-NKWVEPMBSA-N Didanosine Chemical compound O1[C@H](CO)CC[C@@H]1N1C(NC=NC2=O)=C2N=C1 BXZVVICBKDXVGW-NKWVEPMBSA-N 0.000 description 1
- 208000002699 Digestive System Neoplasms Diseases 0.000 description 1
- 102100022817 Disintegrin and metalloproteinase domain-containing protein 29 Human genes 0.000 description 1
- 102100035424 DnaJ homolog subfamily B member 8 Human genes 0.000 description 1
- 102100030068 Doublesex- and mab-3-related transcription factor 1 Human genes 0.000 description 1
- 102100032484 Down syndrome critical region protein 8 Human genes 0.000 description 1
- 241001319090 Dracunculus medinensis Species 0.000 description 1
- 102100036109 Dual specificity protein kinase TTK Human genes 0.000 description 1
- 101100408377 Dugesia japonica iwi gene Proteins 0.000 description 1
- 229930193152 Dynemicin Natural products 0.000 description 1
- 102100035272 E3 ubiquitin-protein ligase CBLL2 Human genes 0.000 description 1
- 108010031111 EBV-encoded nuclear antigen 1 Proteins 0.000 description 1
- 101150029707 ERBB2 gene Proteins 0.000 description 1
- 241000244170 Echinococcus granulosus Species 0.000 description 1
- 102100029722 Ectonucleoside triphosphate diphosphohydrolase 1 Human genes 0.000 description 1
- XPOQHMRABVBWPR-UHFFFAOYSA-N Efavirenz Natural products O1C(=O)NC2=CC=C(Cl)C=C2C1(C(F)(F)F)C#CC1CC1 XPOQHMRABVBWPR-UHFFFAOYSA-N 0.000 description 1
- 206010014418 Electrolyte imbalance Diseases 0.000 description 1
- 102100032053 Elongation of very long chain fatty acids protein 4 Human genes 0.000 description 1
- XQSPYNMVSIKCOC-NTSWFWBYSA-N Emtricitabine Chemical compound C1=C(F)C(N)=NC(=O)N1[C@H]1O[C@@H](CO)SC1 XQSPYNMVSIKCOC-NTSWFWBYSA-N 0.000 description 1
- AFMYMMXSQGUCBK-UHFFFAOYSA-N Endynamicin A Natural products C1#CC=CC#CC2NC(C=3C(=O)C4=C(O)C=CC(O)=C4C(=O)C=3C(O)=C3)=C3C34OC32C(C)C(C(O)=O)=C(OC)C41 AFMYMMXSQGUCBK-UHFFFAOYSA-N 0.000 description 1
- 108010032976 Enfuvirtide Proteins 0.000 description 1
- SAMRUMKYXPVKPA-VFKOLLTISA-N Enocitabine Chemical compound O=C1N=C(NC(=O)CCCCCCCCCCCCCCCCCCCCC)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 SAMRUMKYXPVKPA-VFKOLLTISA-N 0.000 description 1
- 241000224432 Entamoeba histolytica Species 0.000 description 1
- 241000588914 Enterobacter Species 0.000 description 1
- 241000588697 Enterobacter cloacae Species 0.000 description 1
- 241000498255 Enterobius vermicularis Species 0.000 description 1
- 241000194032 Enterococcus faecalis Species 0.000 description 1
- 241000194031 Enterococcus faecium Species 0.000 description 1
- 241000709661 Enterovirus Species 0.000 description 1
- 241000991587 Enterovirus C Species 0.000 description 1
- 206010014967 Ependymoma Diseases 0.000 description 1
- YQYJSBFKSSDGFO-UHFFFAOYSA-N Epihygromycin Natural products OC1C(O)C(C(=O)C)OC1OC(C(=C1)O)=CC=C1C=C(C)C(=O)NC1C(O)C(O)C2OCOC2C1O YQYJSBFKSSDGFO-UHFFFAOYSA-N 0.000 description 1
- HTIJFSOGRVMCQR-UHFFFAOYSA-N Epirubicin Natural products COc1cccc2C(=O)c3c(O)c4CC(O)(CC(OC5CC(N)C(=O)C(C)O5)c4c(O)c3C(=O)c12)C(=O)CO HTIJFSOGRVMCQR-UHFFFAOYSA-N 0.000 description 1
- 108010066687 Epithelial Cell Adhesion Molecule Proteins 0.000 description 1
- 102000018651 Epithelial Cell Adhesion Molecule Human genes 0.000 description 1
- OBMLHUPNRURLOK-XGRAFVIBSA-N Epitiostanol Chemical compound C1[C@@H]2S[C@@H]2C[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CC[C@H]21 OBMLHUPNRURLOK-XGRAFVIBSA-N 0.000 description 1
- 102100031938 Eppin Human genes 0.000 description 1
- 101710122227 Epstein-Barr nuclear antigen 1 Proteins 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 241000283073 Equus caballus Species 0.000 description 1
- 241000588722 Escherichia Species 0.000 description 1
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 1
- 229930189413 Esperamicin Natural products 0.000 description 1
- JOYRKODLDBILNP-UHFFFAOYSA-N Ethyl urethane Chemical compound CCOC(N)=O JOYRKODLDBILNP-UHFFFAOYSA-N 0.000 description 1
- 241000204939 Fasciola gigantica Species 0.000 description 1
- 102100038652 Ferritin heavy polypeptide-like 17 Human genes 0.000 description 1
- 108010069446 Fertilins Proteins 0.000 description 1
- 102000001133 Fertilins Human genes 0.000 description 1
- 102100027603 Fetal and adult testis-expressed transcript protein Human genes 0.000 description 1
- 201000008808 Fibrosarcoma Diseases 0.000 description 1
- 102100027581 Forkhead box protein P3 Human genes 0.000 description 1
- 241000589602 Francisella tularensis Species 0.000 description 1
- 206010017533 Fungal infection Diseases 0.000 description 1
- 102100039717 G antigen 1 Human genes 0.000 description 1
- 102100036295 G antigen 12F Human genes 0.000 description 1
- 102100036299 G antigen 12G Human genes 0.000 description 1
- 102100036298 G antigen 12H Human genes 0.000 description 1
- 102100021019 G antigen 12J Human genes 0.000 description 1
- 102100039712 G antigen 13 Human genes 0.000 description 1
- 102100039709 G antigen 2A Human genes 0.000 description 1
- 101710098476 G antigen 2D Proteins 0.000 description 1
- 102100039700 G antigen 2E Human genes 0.000 description 1
- 102100039699 G antigen 4 Human genes 0.000 description 1
- 102100039698 G antigen 5 Human genes 0.000 description 1
- 102100039713 G antigen 6 Human genes 0.000 description 1
- 102100039805 G patch domain-containing protein 2 Human genes 0.000 description 1
- 101710121810 Galectin-9 Proteins 0.000 description 1
- 101100229077 Gallus gallus GAL9 gene Proteins 0.000 description 1
- 102100024375 Gamma-glutamylaminecyclotransferase Human genes 0.000 description 1
- 101710201613 Gamma-glutamylaminecyclotransferase Proteins 0.000 description 1
- 102100030525 Gap junction alpha-4 protein Human genes 0.000 description 1
- 241000207201 Gardnerella vaginalis Species 0.000 description 1
- 206010017993 Gastrointestinal neoplasms Diseases 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000224467 Giardia intestinalis Species 0.000 description 1
- 206010018338 Glioma Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102100024015 Glycerol-3-phosphate acyltransferase 2, mitochondrial Human genes 0.000 description 1
- 102000007390 Glycogen Phosphorylase Human genes 0.000 description 1
- 108010046163 Glycogen Phosphorylase Proteins 0.000 description 1
- 102000001398 Granzyme Human genes 0.000 description 1
- 108060005986 Granzyme Proteins 0.000 description 1
- 206010053759 Growth retardation Diseases 0.000 description 1
- 102100035943 HERV-H LTR-associating protein 2 Human genes 0.000 description 1
- 208000031886 HIV Infections Diseases 0.000 description 1
- 102100030595 HLA class II histocompatibility antigen gamma chain Human genes 0.000 description 1
- 229940121710 HMGCoA reductase inhibitor Drugs 0.000 description 1
- 102100028673 HORMA domain-containing protein 1 Human genes 0.000 description 1
- 102100028670 HORMA domain-containing protein 2 Human genes 0.000 description 1
- 101150000613 HSPB9 gene Proteins 0.000 description 1
- 206010066476 Haematological malignancy Diseases 0.000 description 1
- 241000606790 Haemophilus Species 0.000 description 1
- 241000606788 Haemophilus haemolyticus Species 0.000 description 1
- 241000606768 Haemophilus influenzae Species 0.000 description 1
- 241000606822 Haemophilus parahaemolyticus Species 0.000 description 1
- 241000606766 Haemophilus parainfluenzae Species 0.000 description 1
- 101100508941 Halobacterium salinarum (strain ATCC 700922 / JCM 11081 / NRC-1) ppa gene Proteins 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- 102100023042 Heat shock protein beta-9 Human genes 0.000 description 1
- 241000590002 Helicobacter pylori Species 0.000 description 1
- 208000006968 Helminthiasis Diseases 0.000 description 1
- 208000002250 Hematologic Neoplasms Diseases 0.000 description 1
- 108091005904 Hemoglobin subunit beta Proteins 0.000 description 1
- 101710083479 Hepatitis A virus cellular receptor 2 homolog Proteins 0.000 description 1
- 208000005176 Hepatitis C Diseases 0.000 description 1
- 241000742052 Heterophyes heterophyes Species 0.000 description 1
- 102000018713 Histocompatibility Antigens Class II Human genes 0.000 description 1
- 241000948220 Histomonas meleagridis Species 0.000 description 1
- 102100031470 Homeobox protein ARX Human genes 0.000 description 1
- 101000678236 Homo sapiens 5'-nucleotidase Proteins 0.000 description 1
- 101000774732 Homo sapiens A-kinase anchor protein 3 Proteins 0.000 description 1
- 101000890604 Homo sapiens A-kinase anchor protein 4 Proteins 0.000 description 1
- 101000796047 Homo sapiens AP-2 complex subunit mu Proteins 0.000 description 1
- 101000887284 Homo sapiens ATPase family AAA domain-containing protein 2 Proteins 0.000 description 1
- 101000756551 Homo sapiens Acrosin-binding protein Proteins 0.000 description 1
- 101000678435 Homo sapiens Actin-like protein 8 Proteins 0.000 description 1
- 101000732375 Homo sapiens Ankyrin repeat domain-containing protein 45 Proteins 0.000 description 1
- 101000684962 Homo sapiens Armadillo repeat-containing protein 3 Proteins 0.000 description 1
- 101000678879 Homo sapiens Atypical chemokine receptor 1 Proteins 0.000 description 1
- 101000874316 Homo sapiens B melanoma antigen 1 Proteins 0.000 description 1
- 101000874318 Homo sapiens B melanoma antigen 2 Proteins 0.000 description 1
- 101000874317 Homo sapiens B melanoma antigen 3 Proteins 0.000 description 1
- 101000874320 Homo sapiens B melanoma antigen 4 Proteins 0.000 description 1
- 101000874319 Homo sapiens B melanoma antigen 5 Proteins 0.000 description 1
- 101000864344 Homo sapiens B- and T-lymphocyte attenuator Proteins 0.000 description 1
- 101000884305 Homo sapiens B-cell receptor CD22 Proteins 0.000 description 1
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 description 1
- 101000794028 Homo sapiens Bromodomain testis-specific protein Proteins 0.000 description 1
- 101001076862 Homo sapiens C-Jun-amino-terminal kinase-interacting protein 4 Proteins 0.000 description 1
- 101000942590 Homo sapiens CCR4-NOT transcription complex subunit 9 Proteins 0.000 description 1
- 101100166600 Homo sapiens CD28 gene Proteins 0.000 description 1
- 101100099884 Homo sapiens CD40 gene Proteins 0.000 description 1
- 101000745609 Homo sapiens CPX chromosomal region candidate gene 1 protein Proteins 0.000 description 1
- 101000935132 Homo sapiens Calcium-binding tyrosine phosphorylation-regulated protein Proteins 0.000 description 1
- 101000741289 Homo sapiens Calreticulin-3 Proteins 0.000 description 1
- 101000933825 Homo sapiens Cancer-associated gene 1 protein Proteins 0.000 description 1
- 101000746248 Homo sapiens Cancer/testis antigen 47B Proteins 0.000 description 1
- 101000940800 Homo sapiens Cancer/testis antigen family 45 member A1 Proteins 0.000 description 1
- 101000940805 Homo sapiens Cancer/testis antigen family 45 member A2 Proteins 0.000 description 1
- 101000940772 Homo sapiens Cancer/testis antigen family 45 member A5 Proteins 0.000 description 1
- 101000940770 Homo sapiens Cancer/testis antigen family 45 member A6 Proteins 0.000 description 1
- 101000859073 Homo sapiens Catenin alpha-2 Proteins 0.000 description 1
- 101000721661 Homo sapiens Cellular tumor antigen p53 Proteins 0.000 description 1
- 101000613559 Homo sapiens Circadian clock protein PASD1 Proteins 0.000 description 1
- 101000868824 Homo sapiens Coiled-coil domain-containing protein 110 Proteins 0.000 description 1
- 101000897106 Homo sapiens Coiled-coil domain-containing protein 33 Proteins 0.000 description 1
- 101000737082 Homo sapiens Coiled-coil domain-containing protein 62 Proteins 0.000 description 1
- 101000932745 Homo sapiens Coiled-coil domain-containing protein 83 Proteins 0.000 description 1
- 101000856239 Homo sapiens Cutaneous T-cell lymphoma-associated antigen 1 Proteins 0.000 description 1
- 101000726255 Homo sapiens Cysteine-rich secretory protein 2 Proteins 0.000 description 1
- 101000922370 Homo sapiens Cytochrome c oxidase subunit 6B2 Proteins 0.000 description 1
- 101000885459 Homo sapiens DDB1- and CUL4-associated factor 12 Proteins 0.000 description 1
- 101000804948 Homo sapiens Developmental pluripotency-associated protein 2 Proteins 0.000 description 1
- 101000951345 Homo sapiens Dickkopf-like protein 1 Proteins 0.000 description 1
- 101000756746 Homo sapiens Disintegrin and metalloproteinase domain-containing protein 29 Proteins 0.000 description 1
- 101000804109 Homo sapiens DnaJ homolog subfamily B member 8 Proteins 0.000 description 1
- 101000864807 Homo sapiens Doublesex- and mab-3-related transcription factor 1 Proteins 0.000 description 1
- 101001016533 Homo sapiens Down syndrome critical region protein 8 Proteins 0.000 description 1
- 101000659223 Homo sapiens Dual specificity protein kinase TTK Proteins 0.000 description 1
- 101000737263 Homo sapiens E3 ubiquitin-protein ligase CBLL2 Proteins 0.000 description 1
- 101001012447 Homo sapiens Ectonucleoside triphosphate diphosphohydrolase 1 Proteins 0.000 description 1
- 101000921354 Homo sapiens Elongation of very long chain fatty acids protein 4 Proteins 0.000 description 1
- 101000920711 Homo sapiens Eppin Proteins 0.000 description 1
- 101001031604 Homo sapiens Ferritin heavy polypeptide-like 17 Proteins 0.000 description 1
- 101000937113 Homo sapiens Fetal and adult testis-expressed transcript protein Proteins 0.000 description 1
- 101000861452 Homo sapiens Forkhead box protein P3 Proteins 0.000 description 1
- 101000886137 Homo sapiens G antigen 1 Proteins 0.000 description 1
- 101001074832 Homo sapiens G antigen 12F Proteins 0.000 description 1
- 101001074830 Homo sapiens G antigen 12G Proteins 0.000 description 1
- 101001074828 Homo sapiens G antigen 12H Proteins 0.000 description 1
- 101001074826 Homo sapiens G antigen 12I Proteins 0.000 description 1
- 101001075398 Homo sapiens G antigen 12J Proteins 0.000 description 1
- 101000886150 Homo sapiens G antigen 13 Proteins 0.000 description 1
- 101000886151 Homo sapiens G antigen 2A Proteins 0.000 description 1
- 101000886136 Homo sapiens G antigen 4 Proteins 0.000 description 1
- 101000886135 Homo sapiens G antigen 5 Proteins 0.000 description 1
- 101000886141 Homo sapiens G antigen 6 Proteins 0.000 description 1
- 101000893968 Homo sapiens G antigen 7 Proteins 0.000 description 1
- 101001034114 Homo sapiens G patch domain-containing protein 2 Proteins 0.000 description 1
- 101000904251 Homo sapiens Glycerol-3-phosphate acyltransferase 2, mitochondrial Proteins 0.000 description 1
- 101001082627 Homo sapiens HLA class II histocompatibility antigen gamma chain Proteins 0.000 description 1
- 101000985274 Homo sapiens HORMA domain-containing protein 1 Proteins 0.000 description 1
- 101000985263 Homo sapiens HORMA domain-containing protein 2 Proteins 0.000 description 1
- 101001068133 Homo sapiens Hepatitis A virus cellular receptor 2 Proteins 0.000 description 1
- 101000923090 Homo sapiens Homeobox protein ARX Proteins 0.000 description 1
- 101001019455 Homo sapiens ICOS ligand Proteins 0.000 description 1
- 101000705915 Homo sapiens Inactive serine protease 54 Proteins 0.000 description 1
- 101000889893 Homo sapiens Inactive serine/threonine-protein kinase TEX14 Proteins 0.000 description 1
- 101001055250 Homo sapiens Interactor of HORMAD1 protein 1 Proteins 0.000 description 1
- 101001057504 Homo sapiens Interferon-stimulated gene 20 kDa protein Proteins 0.000 description 1
- 101001003132 Homo sapiens Interleukin-13 receptor subunit alpha-2 Proteins 0.000 description 1
- 101001055144 Homo sapiens Interleukin-2 receptor subunit alpha Proteins 0.000 description 1
- 101000590482 Homo sapiens Kinetochore protein Nuf2 Proteins 0.000 description 1
- 101000971521 Homo sapiens Kinetochore scaffold 1 Proteins 0.000 description 1
- 101001130171 Homo sapiens L-lactate dehydrogenase C chain Proteins 0.000 description 1
- 101001018097 Homo sapiens L-selectin Proteins 0.000 description 1
- 101001007415 Homo sapiens LEM domain-containing protein 1 Proteins 0.000 description 1
- 101001054842 Homo sapiens Leucine zipper protein 4 Proteins 0.000 description 1
- 101000777628 Homo sapiens Leukocyte antigen CD37 Proteins 0.000 description 1
- 101001137987 Homo sapiens Lymphocyte activation gene 3 protein Proteins 0.000 description 1
- 101001065550 Homo sapiens Lymphocyte antigen 6K Proteins 0.000 description 1
- 101000762967 Homo sapiens Lymphokine-activated killer T-cell-originated protein kinase Proteins 0.000 description 1
- 101000578784 Homo sapiens Melanoma antigen recognized by T-cells 1 Proteins 0.000 description 1
- 101001012669 Homo sapiens Melanoma inhibitory activity protein 2 Proteins 0.000 description 1
- 101001005728 Homo sapiens Melanoma-associated antigen 1 Proteins 0.000 description 1
- 101001005725 Homo sapiens Melanoma-associated antigen 10 Proteins 0.000 description 1
- 101001005716 Homo sapiens Melanoma-associated antigen 11 Proteins 0.000 description 1
- 101001005717 Homo sapiens Melanoma-associated antigen 12 Proteins 0.000 description 1
- 101001005719 Homo sapiens Melanoma-associated antigen 3 Proteins 0.000 description 1
- 101001005720 Homo sapiens Melanoma-associated antigen 4 Proteins 0.000 description 1
- 101001005722 Homo sapiens Melanoma-associated antigen 6 Proteins 0.000 description 1
- 101001005723 Homo sapiens Melanoma-associated antigen 8 Proteins 0.000 description 1
- 101001036688 Homo sapiens Melanoma-associated antigen B1 Proteins 0.000 description 1
- 101001036686 Homo sapiens Melanoma-associated antigen B2 Proteins 0.000 description 1
- 101001036692 Homo sapiens Melanoma-associated antigen B3 Proteins 0.000 description 1
- 101001036691 Homo sapiens Melanoma-associated antigen B4 Proteins 0.000 description 1
- 101001036689 Homo sapiens Melanoma-associated antigen B5 Proteins 0.000 description 1
- 101001036675 Homo sapiens Melanoma-associated antigen B6 Proteins 0.000 description 1
- 101001036406 Homo sapiens Melanoma-associated antigen C1 Proteins 0.000 description 1
- 101001057156 Homo sapiens Melanoma-associated antigen C2 Proteins 0.000 description 1
- 101001057159 Homo sapiens Melanoma-associated antigen C3 Proteins 0.000 description 1
- 101001133056 Homo sapiens Mucin-1 Proteins 0.000 description 1
- 101000934338 Homo sapiens Myeloid cell surface antigen CD33 Proteins 0.000 description 1
- 101001128135 Homo sapiens NACHT, LRR and PYD domains-containing protein 4 Proteins 0.000 description 1
- 101000884270 Homo sapiens Natural killer cell receptor 2B4 Proteins 0.000 description 1
- 101001109682 Homo sapiens Nuclear receptor subfamily 6 group A member 1 Proteins 0.000 description 1
- 101001134172 Homo sapiens Otoancorin Proteins 0.000 description 1
- 101001114057 Homo sapiens P antigen family member 1 Proteins 0.000 description 1
- 101001114056 Homo sapiens P antigen family member 2 Proteins 0.000 description 1
- 101001114053 Homo sapiens P antigen family member 3 Proteins 0.000 description 1
- 101000741879 Homo sapiens POTE ankyrin domain family member A Proteins 0.000 description 1
- 101000741880 Homo sapiens POTE ankyrin domain family member B Proteins 0.000 description 1
- 101000741895 Homo sapiens POTE ankyrin domain family member C Proteins 0.000 description 1
- 101000741896 Homo sapiens POTE ankyrin domain family member D Proteins 0.000 description 1
- 101000741893 Homo sapiens POTE ankyrin domain family member E Proteins 0.000 description 1
- 101000741899 Homo sapiens POTE ankyrin domain family member G Proteins 0.000 description 1
- 101000741900 Homo sapiens POTE ankyrin domain family member H Proteins 0.000 description 1
- 101001064774 Homo sapiens Peroxidasin-like protein Proteins 0.000 description 1
- 101001126084 Homo sapiens Piwi-like protein 2 Proteins 0.000 description 1
- 101000691463 Homo sapiens Placenta-specific protein 1 Proteins 0.000 description 1
- 101001096178 Homo sapiens Pleckstrin homology domain-containing family A member 5 Proteins 0.000 description 1
- 101000874141 Homo sapiens Probable ATP-dependent RNA helicase DDX43 Proteins 0.000 description 1
- 101000883798 Homo sapiens Probable ATP-dependent RNA helicase DDX53 Proteins 0.000 description 1
- 101001100332 Homo sapiens Probable RNA-binding protein 46 Proteins 0.000 description 1
- 101000633613 Homo sapiens Probable threonine protease PRSS50 Proteins 0.000 description 1
- 101001090148 Homo sapiens Protamine-2 Proteins 0.000 description 1
- 101000882139 Homo sapiens Protein FAM133A Proteins 0.000 description 1
- 101000880769 Homo sapiens Protein SSX1 Proteins 0.000 description 1
- 101000880771 Homo sapiens Protein SSX3 Proteins 0.000 description 1
- 101000880775 Homo sapiens Protein SSX5 Proteins 0.000 description 1
- 101000880773 Homo sapiens Protein SSX7 Proteins 0.000 description 1
- 101000956414 Homo sapiens Protein maelstrom homolog Proteins 0.000 description 1
- 101000886682 Homo sapiens Putative G antigen family E member 3 Proteins 0.000 description 1
- 101000745415 Homo sapiens Putative chondrosarcoma-associated gene 1 protein Proteins 0.000 description 1
- 101001005721 Homo sapiens Putative melanoma-associated antigen 5P Proteins 0.000 description 1
- 101000880772 Homo sapiens Putative protein SSX6 Proteins 0.000 description 1
- 101000642817 Homo sapiens Putative protein SSX9 Proteins 0.000 description 1
- 101000725916 Homo sapiens Putative tumor antigen NA88-A Proteins 0.000 description 1
- 101000679365 Homo sapiens Putative tyrosine-protein phosphatase TPTE Proteins 0.000 description 1
- 101000712009 Homo sapiens RING finger protein 17 Proteins 0.000 description 1
- 101001062222 Homo sapiens Receptor-binding cancer antigen expressed on SiSo cells Proteins 0.000 description 1
- 101001095435 Homo sapiens Rhox homeobox family member 2 Proteins 0.000 description 1
- 101001088125 Homo sapiens Ropporin-1A Proteins 0.000 description 1
- 101000821981 Homo sapiens Sarcoma antigen 1 Proteins 0.000 description 1
- 101000739754 Homo sapiens Semenogelin-1 Proteins 0.000 description 1
- 101000705953 Homo sapiens Serine protease 55 Proteins 0.000 description 1
- 101000701401 Homo sapiens Serine/threonine-protein kinase 38 Proteins 0.000 description 1
- 101000777277 Homo sapiens Serine/threonine-protein kinase Chk2 Proteins 0.000 description 1
- 101000785887 Homo sapiens Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit alpha isoform Proteins 0.000 description 1
- 101000637373 Homo sapiens Sperm acrosome membrane-associated protein 3 Proteins 0.000 description 1
- 101000702102 Homo sapiens Sperm flagellar protein 2 Proteins 0.000 description 1
- 101001038163 Homo sapiens Sperm protamine P1 Proteins 0.000 description 1
- 101000825248 Homo sapiens Sperm protein associated with the nucleus on the X chromosome C Proteins 0.000 description 1
- 101000587782 Homo sapiens Sperm protein associated with the nucleus on the X chromosome N1 Proteins 0.000 description 1
- 101000651400 Homo sapiens Sperm protein associated with the nucleus on the X chromosome N2 Proteins 0.000 description 1
- 101000651402 Homo sapiens Sperm protein associated with the nucleus on the X chromosome N3 Proteins 0.000 description 1
- 101000651404 Homo sapiens Sperm protein associated with the nucleus on the X chromosome N4 Proteins 0.000 description 1
- 101000651405 Homo sapiens Sperm protein associated with the nucleus on the X chromosome N5 Proteins 0.000 description 1
- 101000824971 Homo sapiens Sperm surface protein Sp17 Proteins 0.000 description 1
- 101000618135 Homo sapiens Sperm-associated antigen 1 Proteins 0.000 description 1
- 101000642433 Homo sapiens Sperm-associated antigen 17 Proteins 0.000 description 1
- 101000618138 Homo sapiens Sperm-associated antigen 4 protein Proteins 0.000 description 1
- 101000618139 Homo sapiens Sperm-associated antigen 6 Proteins 0.000 description 1
- 101000618112 Homo sapiens Sperm-associated antigen 8 Proteins 0.000 description 1
- 101000642323 Homo sapiens Spermatogenesis-associated protein 19, mitochondrial Proteins 0.000 description 1
- 101000828537 Homo sapiens Synaptic functional regulator FMR1 Proteins 0.000 description 1
- 101000630833 Homo sapiens Synaptonemal complex central element protein 1 Proteins 0.000 description 1
- 101000643620 Homo sapiens Synaptonemal complex protein 1 Proteins 0.000 description 1
- 101000874179 Homo sapiens Syndecan-1 Proteins 0.000 description 1
- 101000831007 Homo sapiens T-cell immunoreceptor with Ig and ITIM domains Proteins 0.000 description 1
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 description 1
- 101100207070 Homo sapiens TNFSF8 gene Proteins 0.000 description 1
- 101000655330 Homo sapiens Tektin-5 Proteins 0.000 description 1
- 101000666389 Homo sapiens Terminal nucleotidyltransferase 5D Proteins 0.000 description 1
- 101000655622 Homo sapiens Testicular haploid expressed gene protein Proteins 0.000 description 1
- 101000795918 Homo sapiens Testis-expressed protein 101 Proteins 0.000 description 1
- 101000596845 Homo sapiens Testis-expressed protein 15 Proteins 0.000 description 1
- 101000759895 Homo sapiens Testis-specific Y-encoded protein 2 Proteins 0.000 description 1
- 101000759894 Homo sapiens Testis-specific Y-encoded protein 3 Proteins 0.000 description 1
- 101000612981 Homo sapiens Testis-specific gene 10 protein Proteins 0.000 description 1
- 101000794200 Homo sapiens Testis-specific serine/threonine-protein kinase 6 Proteins 0.000 description 1
- 101000834937 Homo sapiens Tomoregulin-1 Proteins 0.000 description 1
- 101000834948 Homo sapiens Tomoregulin-2 Proteins 0.000 description 1
- 101000666379 Homo sapiens Transcription factor Dp family member 3 Proteins 0.000 description 1
- 101000674717 Homo sapiens Transcription initiation factor TFIID subunit 7-like Proteins 0.000 description 1
- 101000894428 Homo sapiens Transcriptional repressor CTCFL Proteins 0.000 description 1
- 101000764622 Homo sapiens Transmembrane and immunoglobulin domain-containing protein 2 Proteins 0.000 description 1
- 101000904724 Homo sapiens Transmembrane glycoprotein NMB Proteins 0.000 description 1
- 101000798715 Homo sapiens Transmembrane protease serine 12 Proteins 0.000 description 1
- 101000852860 Homo sapiens Transmembrane protein 108 Proteins 0.000 description 1
- 101000772169 Homo sapiens Tubby-related protein 2 Proteins 0.000 description 1
- 101000637036 Homo sapiens Tubulin polymerization-promoting protein family member 2 Proteins 0.000 description 1
- 101000889756 Homo sapiens Tudor domain-containing protein 1 Proteins 0.000 description 1
- 101000835790 Homo sapiens Tudor domain-containing protein 6 Proteins 0.000 description 1
- 101000830594 Homo sapiens Tumor necrosis factor ligand superfamily member 14 Proteins 0.000 description 1
- 101000638255 Homo sapiens Tumor necrosis factor ligand superfamily member 8 Proteins 0.000 description 1
- 101000648507 Homo sapiens Tumor necrosis factor receptor superfamily member 14 Proteins 0.000 description 1
- 101001087404 Homo sapiens Tyrosine-protein phosphatase non-receptor type 20 Proteins 0.000 description 1
- 101000863873 Homo sapiens Tyrosine-protein phosphatase non-receptor type substrate 1 Proteins 0.000 description 1
- 101000814497 Homo sapiens X antigen family member 3 Proteins 0.000 description 1
- 101000814496 Homo sapiens X antigen family member 5 Proteins 0.000 description 1
- 101000964582 Homo sapiens Zinc finger protein 165 Proteins 0.000 description 1
- 101000856240 Homo sapiens cTAGE family member 2 Proteins 0.000 description 1
- 241001213909 Human endogenous retroviruses Species 0.000 description 1
- 241000342334 Human metapneumovirus Species 0.000 description 1
- 241000701806 Human papillomavirus Species 0.000 description 1
- 101100321817 Human parvovirus B19 (strain HV) 7.5K gene Proteins 0.000 description 1
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 description 1
- VSNHCAURESNICA-UHFFFAOYSA-N Hydroxyurea Chemical compound NC(=O)NO VSNHCAURESNICA-UHFFFAOYSA-N 0.000 description 1
- 241001464384 Hymenolepis nana Species 0.000 description 1
- 206010020631 Hypergammaglobulinaemia benign monoclonal Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 108010091358 Hypoxanthine Phosphoribosyltransferase Proteins 0.000 description 1
- 102100029098 Hypoxanthine-guanine phosphoribosyltransferase Human genes 0.000 description 1
- 101150112877 IGSF11 gene Proteins 0.000 description 1
- MPBVHIBUJCELCL-UHFFFAOYSA-N Ibandronate Chemical compound CCCCCN(C)CCC(O)(P(O)(O)=O)P(O)(O)=O MPBVHIBUJCELCL-UHFFFAOYSA-N 0.000 description 1
- XDXDZDZNSLXDNA-TZNDIEGXSA-N Idarubicin Chemical compound C1[C@H](N)[C@H](O)[C@H](C)O[C@H]1O[C@@H]1C2=C(O)C(C(=O)C3=CC=CC=C3C3=O)=C3C(O)=C2C[C@@](O)(C(C)=O)C1 XDXDZDZNSLXDNA-TZNDIEGXSA-N 0.000 description 1
- XDXDZDZNSLXDNA-UHFFFAOYSA-N Idarubicin Natural products C1C(N)C(O)C(C)OC1OC1C2=C(O)C(C(=O)C3=CC=CC=C3C3=O)=C3C(O)=C2CC(O)(C(C)=O)C1 XDXDZDZNSLXDNA-UHFFFAOYSA-N 0.000 description 1
- 102000037982 Immune checkpoint proteins Human genes 0.000 description 1
- 108091008036 Immune checkpoint proteins Proteins 0.000 description 1
- 208000029462 Immunodeficiency disease Diseases 0.000 description 1
- 102000018071 Immunoglobulin Fc Fragments Human genes 0.000 description 1
- 108010091135 Immunoglobulin Fc Fragments Proteins 0.000 description 1
- 108010043496 Immunoglobulin Idiotypes Proteins 0.000 description 1
- 102100029572 Immunoglobulin kappa constant Human genes 0.000 description 1
- 108010090227 Immunoglobulin kappa-Chains Proteins 0.000 description 1
- 102100021032 Immunoglobulin superfamily member 11 Human genes 0.000 description 1
- 102100031071 Inactive serine protease 54 Human genes 0.000 description 1
- 102100040173 Inactive serine/threonine-protein kinase TEX14 Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 102100026213 Interactor of HORMAD1 protein 1 Human genes 0.000 description 1
- 108010064593 Intercellular Adhesion Molecule-1 Proteins 0.000 description 1
- 102100037877 Intercellular adhesion molecule 1 Human genes 0.000 description 1
- 102100037850 Interferon gamma Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 102000003814 Interleukin-10 Human genes 0.000 description 1
- 108090000174 Interleukin-10 Proteins 0.000 description 1
- 108010065805 Interleukin-12 Proteins 0.000 description 1
- 102000013462 Interleukin-12 Human genes 0.000 description 1
- 102100020793 Interleukin-13 receptor subunit alpha-2 Human genes 0.000 description 1
- 102000013691 Interleukin-17 Human genes 0.000 description 1
- 108050003558 Interleukin-17 Proteins 0.000 description 1
- 102000000588 Interleukin-2 Human genes 0.000 description 1
- 108010002350 Interleukin-2 Proteins 0.000 description 1
- 108010038453 Interleukin-2 Receptors Proteins 0.000 description 1
- 102000010789 Interleukin-2 Receptors Human genes 0.000 description 1
- 102100026878 Interleukin-2 receptor subunit alpha Human genes 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 102000004388 Interleukin-4 Human genes 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 102000004889 Interleukin-6 Human genes 0.000 description 1
- JUZNIMUFDBIJCM-ANEDZVCMSA-N Invanz Chemical compound O=C([C@H]1NC[C@H](C1)SC=1[C@H](C)[C@@H]2[C@H](C(N2C=1C(O)=O)=O)[C@H](O)C)NC1=CC=CC(C(O)=O)=C1 JUZNIMUFDBIJCM-ANEDZVCMSA-N 0.000 description 1
- 241000710842 Japanese encephalitis virus Species 0.000 description 1
- 101710015718 KIAA0100 Proteins 0.000 description 1
- 102000002698 KIR Receptors Human genes 0.000 description 1
- 108010043610 KIR Receptors Proteins 0.000 description 1
- 108010025815 Kanamycin Kinase Proteins 0.000 description 1
- 102100023424 Kinesin-like protein KIF2C Human genes 0.000 description 1
- 101710134369 Kinesin-like protein KIF2C Proteins 0.000 description 1
- 102100032431 Kinetochore protein Nuf2 Human genes 0.000 description 1
- 102100021464 Kinetochore scaffold 1 Human genes 0.000 description 1
- 241001454354 Kingella Species 0.000 description 1
- 241000588748 Klebsiella Species 0.000 description 1
- 241000588915 Klebsiella aerogenes Species 0.000 description 1
- 241000588749 Klebsiella oxytoca Species 0.000 description 1
- 241000588747 Klebsiella pneumoniae Species 0.000 description 1
- SNDPXSYFESPGGJ-BYPYZUCNSA-N L-2-aminopentanoic acid Chemical compound CCC[C@H](N)C(O)=O SNDPXSYFESPGGJ-BYPYZUCNSA-N 0.000 description 1
- AHLPHDHHMVZTML-BYPYZUCNSA-N L-Ornithine Chemical compound NCCC[C@H](N)C(O)=O AHLPHDHHMVZTML-BYPYZUCNSA-N 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- ZGUNAGUHMKGQNY-ZETCQYMHSA-N L-alpha-phenylglycine zwitterion Chemical compound OC(=O)[C@@H](N)C1=CC=CC=C1 ZGUNAGUHMKGQNY-ZETCQYMHSA-N 0.000 description 1
- RHGKLRLOHDJJDR-BYPYZUCNSA-N L-citrulline Chemical compound NC(=O)NCCC[C@H]([NH3+])C([O-])=O RHGKLRLOHDJJDR-BYPYZUCNSA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- XIGSAGMEBXLVJJ-YFKPBYRVSA-N L-homocitrulline Chemical compound NC(=O)NCCCC[C@H]([NH3+])C([O-])=O XIGSAGMEBXLVJJ-YFKPBYRVSA-N 0.000 description 1
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 1
- 102100031357 L-lactate dehydrogenase C chain Human genes 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- SNDPXSYFESPGGJ-UHFFFAOYSA-N L-norVal-OH Natural products CCCC(N)C(O)=O SNDPXSYFESPGGJ-UHFFFAOYSA-N 0.000 description 1
- LRQKBLKVPFOOQJ-YFKPBYRVSA-N L-norleucine Chemical compound CCCC[C@H]([NH3+])C([O-])=O LRQKBLKVPFOOQJ-YFKPBYRVSA-N 0.000 description 1
- ZFOMKMMPBOQKMC-KXUCPTDWSA-N L-pyrrolysine Chemical compound C[C@@H]1CC=N[C@H]1C(=O)NCCCC[C@H]([NH3+])C([O-])=O ZFOMKMMPBOQKMC-KXUCPTDWSA-N 0.000 description 1
- 102100033467 L-selectin Human genes 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- 102100028300 LEM domain-containing protein 1 Human genes 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 101150030213 Lag3 gene Proteins 0.000 description 1
- 206010023825 Laryngeal cancer Diseases 0.000 description 1
- 241000589242 Legionella pneumophila Species 0.000 description 1
- 241000222740 Leishmania braziliensis Species 0.000 description 1
- 241000222727 Leishmania donovani Species 0.000 description 1
- 241000222734 Leishmania mexicana Species 0.000 description 1
- 241000222736 Leishmania tropica Species 0.000 description 1
- 229920001491 Lentinan Polymers 0.000 description 1
- 241000713666 Lentivirus Species 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 102100026910 Leucine zipper protein 4 Human genes 0.000 description 1
- 102100031586 Leukocyte antigen CD37 Human genes 0.000 description 1
- GSDSWSVVBLHKDQ-JTQLQIEISA-N Levofloxacin Chemical compound C([C@@H](N1C2=C(C(C(C(O)=O)=C1)=O)C=C1F)C)OC2=C1N1CCN(C)CC1 GSDSWSVVBLHKDQ-JTQLQIEISA-N 0.000 description 1
- 208000006552 Lewis Lung Carcinoma Diseases 0.000 description 1
- 101710102461 Lipase member I Proteins 0.000 description 1
- 102100030659 Lipase member I Human genes 0.000 description 1
- 241000186779 Listeria monocytogenes Species 0.000 description 1
- GQYIWUVLTXOXAJ-UHFFFAOYSA-N Lomustine Chemical compound ClCCN(N=O)C(=O)NC1CCCCC1 GQYIWUVLTXOXAJ-UHFFFAOYSA-N 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 102100032129 Lymphocyte antigen 6K Human genes 0.000 description 1
- 102100026753 Lymphokine-activated killer T-cell-originated protein kinase Human genes 0.000 description 1
- 206010025312 Lymphoma AIDS related Diseases 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 108010010995 MART-1 Antigen Proteins 0.000 description 1
- 108091054438 MHC class II family Proteins 0.000 description 1
- 241001293418 Mannheimia haemolytica Species 0.000 description 1
- 208000025205 Mantle-Cell Lymphoma Diseases 0.000 description 1
- VJRAUFKOOPNFIQ-UHFFFAOYSA-N Marcellomycin Natural products C12=C(O)C=3C(=O)C4=C(O)C=CC(O)=C4C(=O)C=3C=C2C(C(=O)OC)C(CC)(O)CC1OC(OC1C)CC(N(C)C)C1OC(OC1C)CC(O)C1OC1CC(O)C(O)C(C)O1 VJRAUFKOOPNFIQ-UHFFFAOYSA-N 0.000 description 1
- 229930126263 Maytansine Natural products 0.000 description 1
- 241000712079 Measles morbillivirus Species 0.000 description 1
- 208000006395 Meigs Syndrome Diseases 0.000 description 1
- 206010027139 Meigs' syndrome Diseases 0.000 description 1
- 102100029778 Melanoma inhibitory activity protein 2 Human genes 0.000 description 1
- 102100025050 Melanoma-associated antigen 1 Human genes 0.000 description 1
- 102100025049 Melanoma-associated antigen 10 Human genes 0.000 description 1
- 102100025083 Melanoma-associated antigen 11 Human genes 0.000 description 1
- 102100025084 Melanoma-associated antigen 12 Human genes 0.000 description 1
- 102100025082 Melanoma-associated antigen 3 Human genes 0.000 description 1
- 102100025077 Melanoma-associated antigen 4 Human genes 0.000 description 1
- 102100025075 Melanoma-associated antigen 6 Human genes 0.000 description 1
- 102100025076 Melanoma-associated antigen 8 Human genes 0.000 description 1
- 102100039477 Melanoma-associated antigen B1 Human genes 0.000 description 1
- 102100039479 Melanoma-associated antigen B2 Human genes 0.000 description 1
- 102100039473 Melanoma-associated antigen B3 Human genes 0.000 description 1
- 102100039476 Melanoma-associated antigen B4 Human genes 0.000 description 1
- 102100039475 Melanoma-associated antigen B5 Human genes 0.000 description 1
- 102100039483 Melanoma-associated antigen B6 Human genes 0.000 description 1
- 102100039447 Melanoma-associated antigen C1 Human genes 0.000 description 1
- 102100027252 Melanoma-associated antigen C2 Human genes 0.000 description 1
- 102100027248 Melanoma-associated antigen C3 Human genes 0.000 description 1
- IVDYZAAPOLNZKG-KWHRADDSSA-N Mepitiostane Chemical compound O([C@@H]1[C@]2(CC[C@@H]3[C@@]4(C)C[C@H]5S[C@H]5C[C@@H]4CC[C@H]3[C@@H]2CC1)C)C1(OC)CCCC1 IVDYZAAPOLNZKG-KWHRADDSSA-N 0.000 description 1
- 102000003792 Metallothionein Human genes 0.000 description 1
- 206010051696 Metastases to meninges Diseases 0.000 description 1
- RJQXTJLFIWVMTO-TYNCELHUSA-N Methicillin Chemical compound COC1=CC=CC(OC)=C1C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 RJQXTJLFIWVMTO-TYNCELHUSA-N 0.000 description 1
- VFKZTMPDYBFSTM-KVTDHHQDSA-N Mitobronitol Chemical compound BrC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CBr VFKZTMPDYBFSTM-KVTDHHQDSA-N 0.000 description 1
- 229930192392 Mitomycin Natural products 0.000 description 1
- 241000713869 Moloney murine leukemia virus Species 0.000 description 1
- 241000588621 Moraxella Species 0.000 description 1
- 241000588655 Moraxella catarrhalis Species 0.000 description 1
- 241000588772 Morganella morganii Species 0.000 description 1
- 102100034256 Mucin-1 Human genes 0.000 description 1
- 241000711386 Mumps virus Species 0.000 description 1
- 101100481763 Mus musculus Tnfsf4 gene Proteins 0.000 description 1
- 101100207071 Mus musculus Tnfsf8 gene Proteins 0.000 description 1
- 241000186367 Mycobacterium avium Species 0.000 description 1
- 101100023550 Mycobacterium bovis (strain ATCC BAA-935 / AF2122/97) cmaD gene Proteins 0.000 description 1
- 241000186364 Mycobacterium intracellulare Species 0.000 description 1
- 241000186362 Mycobacterium leprae Species 0.000 description 1
- 241000187481 Mycobacterium phlei Species 0.000 description 1
- 241000187479 Mycobacterium tuberculosis Species 0.000 description 1
- 208000031888 Mycoses Diseases 0.000 description 1
- 101710107068 Myelin basic protein Proteins 0.000 description 1
- 102100025243 Myeloid cell surface antigen CD33 Human genes 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 description 1
- PYUSHNKNPOHWEZ-YFKPBYRVSA-N N-formyl-L-methionine Chemical compound CSCC[C@@H](C(O)=O)NC=O PYUSHNKNPOHWEZ-YFKPBYRVSA-N 0.000 description 1
- 102100031898 NACHT, LRR and PYD domains-containing protein 4 Human genes 0.000 description 1
- 101000942113 Naja naja Cysteine-rich venom protein Proteins 0.000 description 1
- 102100029527 Natural cytotoxicity triggering receptor 3 ligand 1 Human genes 0.000 description 1
- 102100038082 Natural killer cell receptor 2B4 Human genes 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- RHGKLRLOHDJJDR-UHFFFAOYSA-N Ndelta-carbamoyl-DL-ornithine Natural products OC(=O)C(N)CCCNC(N)=O RHGKLRLOHDJJDR-UHFFFAOYSA-N 0.000 description 1
- 241000498270 Necator americanus Species 0.000 description 1
- 241000588652 Neisseria gonorrhoeae Species 0.000 description 1
- 241000588650 Neisseria meningitidis Species 0.000 description 1
- 241000244206 Nematoda Species 0.000 description 1
- 208000003788 Neoplasm Micrometastasis Diseases 0.000 description 1
- 206010029260 Neuroblastoma Diseases 0.000 description 1
- 201000004404 Neurofibroma Diseases 0.000 description 1
- 208000005890 Neuroma Diseases 0.000 description 1
- SYNHCENRCUAUNM-UHFFFAOYSA-N Nitrogen mustard N-oxide hydrochloride Chemical compound Cl.ClCC[N+]([O-])(C)CCCl SYNHCENRCUAUNM-UHFFFAOYSA-N 0.000 description 1
- KGTDRFCXGRULNK-UHFFFAOYSA-N Nogalamycin Natural products COC1C(OC)(C)C(OC)C(C)OC1OC1C2=C(O)C(C(=O)C3=C(O)C=C4C5(C)OC(C(C(C5O)N(C)C)O)OC4=C3C3=O)=C3C=C2C(C(=O)OC)C(C)(O)C1 KGTDRFCXGRULNK-UHFFFAOYSA-N 0.000 description 1
- 108010051791 Nuclear Antigens Proteins 0.000 description 1
- 102000019040 Nuclear Antigens Human genes 0.000 description 1
- 102100022670 Nuclear receptor subfamily 6 group A member 1 Human genes 0.000 description 1
- 241001135232 Odoribacter splanchnicus Species 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 1
- 201000010133 Oligodendroglioma Diseases 0.000 description 1
- 229930187135 Olivomycin Natural products 0.000 description 1
- AHLPHDHHMVZTML-UHFFFAOYSA-N Orn-delta-NH2 Natural products NCCCC(N)C(O)=O AHLPHDHHMVZTML-UHFFFAOYSA-N 0.000 description 1
- UTJLXEIPEHZYQJ-UHFFFAOYSA-N Ornithine Natural products OC(=O)C(C)CCCN UTJLXEIPEHZYQJ-UHFFFAOYSA-N 0.000 description 1
- 241000150452 Orthohantavirus Species 0.000 description 1
- 241000712464 Orthomyxoviridae Species 0.000 description 1
- 102100034199 Otoancorin Human genes 0.000 description 1
- 239000004100 Oxytetracycline Substances 0.000 description 1
- 102100023219 P antigen family member 1 Human genes 0.000 description 1
- 102100023220 P antigen family member 2 Human genes 0.000 description 1
- 102100023239 P antigen family member 3 Human genes 0.000 description 1
- 102100038749 POTE ankyrin domain family member A Human genes 0.000 description 1
- 102100038746 POTE ankyrin domain family member B Human genes 0.000 description 1
- 102100038763 POTE ankyrin domain family member C Human genes 0.000 description 1
- 102100038762 POTE ankyrin domain family member D Human genes 0.000 description 1
- 102100038761 POTE ankyrin domain family member E Human genes 0.000 description 1
- 102100038759 POTE ankyrin domain family member G Human genes 0.000 description 1
- 102100038758 POTE ankyrin domain family member H Human genes 0.000 description 1
- 102000036673 PRAME Human genes 0.000 description 1
- 108060006580 PRAME Proteins 0.000 description 1
- 102100034640 PWWP domain-containing DNA repair factor 3A Human genes 0.000 description 1
- 108050007154 PWWP domain-containing DNA repair factor 3A Proteins 0.000 description 1
- 241000282577 Pan troglodytes Species 0.000 description 1
- VREZDOWOLGNDPW-MYVCAWNPSA-N Pancratistatin Natural products O=C1N[C@H]2[C@H](O)[C@H](O)[C@H](O)[C@H](O)[C@@H]2c2c1c(O)c1OCOc1c2 VREZDOWOLGNDPW-MYVCAWNPSA-N 0.000 description 1
- VREZDOWOLGNDPW-ALTGWBOUSA-N Pancratistatin Chemical compound C1=C2[C@H]3[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O)[C@@H]3NC(=O)C2=C(O)C2=C1OCO2 VREZDOWOLGNDPW-ALTGWBOUSA-N 0.000 description 1
- 108010067372 Pancreatic elastase Proteins 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- 208000009608 Papillomavirus Infections Diseases 0.000 description 1
- 241001504519 Papio ursinus Species 0.000 description 1
- 241000606210 Parabacteroides distasonis Species 0.000 description 1
- 241001480234 Paragonimus westermani Species 0.000 description 1
- 241000711504 Paramyxoviridae Species 0.000 description 1
- 208000002606 Paramyxoviridae Infections Diseases 0.000 description 1
- 241000606856 Pasteurella multocida Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 229930195708 Penicillin V Natural products 0.000 description 1
- 108010057150 Peplomycin Proteins 0.000 description 1
- 102100040283 Peptidyl-prolyl cis-trans isomerase B Human genes 0.000 description 1
- 241000206591 Peptococcus Species 0.000 description 1
- 241000150350 Peribunyaviridae Species 0.000 description 1
- 102100031894 Peroxidasin-like protein Human genes 0.000 description 1
- 206010048734 Phakomatosis Diseases 0.000 description 1
- 241000709664 Picornaviridae Species 0.000 description 1
- KMSKQZKKOZQFFG-HSUXVGOQSA-N Pirarubicin Chemical compound O([C@H]1[C@@H](N)C[C@@H](O[C@H]1C)O[C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1CCCCO1 KMSKQZKKOZQFFG-HSUXVGOQSA-N 0.000 description 1
- 208000007913 Pituitary Neoplasms Diseases 0.000 description 1
- 201000005746 Pituitary adenoma Diseases 0.000 description 1
- 206010061538 Pituitary tumour benign Diseases 0.000 description 1
- 102100029365 Piwi-like protein 2 Human genes 0.000 description 1
- 102100026181 Placenta-specific protein 1 Human genes 0.000 description 1
- 208000007452 Plasmacytoma Diseases 0.000 description 1
- 241000223960 Plasmodium falciparum Species 0.000 description 1
- 241000223821 Plasmodium malariae Species 0.000 description 1
- 241001505293 Plasmodium ovale Species 0.000 description 1
- 241000223810 Plasmodium vivax Species 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 208000037062 Polyps Diseases 0.000 description 1
- HFVNWDWLWUCIHC-GUPDPFMOSA-N Prednimustine Chemical compound O=C([C@@]1(O)CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)[C@@H](O)C[C@@]21C)COC(=O)CCCC1=CC=C(N(CCCl)CCCl)C=C1 HFVNWDWLWUCIHC-GUPDPFMOSA-N 0.000 description 1
- 102100035724 Probable ATP-dependent RNA helicase DDX43 Human genes 0.000 description 1
- 102100038236 Probable ATP-dependent RNA helicase DDX53 Human genes 0.000 description 1
- 102100038818 Probable RNA-binding protein 46 Human genes 0.000 description 1
- 102100029523 Probable threonine protease PRSS50 Human genes 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 102100034750 Protamine-2 Human genes 0.000 description 1
- 102100038988 Protein FAM133A Human genes 0.000 description 1
- 108010029485 Protein Isoforms Proteins 0.000 description 1
- 102000001708 Protein Isoforms Human genes 0.000 description 1
- 102100037163 Protein KIAA0100 Human genes 0.000 description 1
- 102100037687 Protein SSX1 Human genes 0.000 description 1
- 102100037726 Protein SSX3 Human genes 0.000 description 1
- 102100037723 Protein SSX5 Human genes 0.000 description 1
- 102100037728 Protein SSX7 Human genes 0.000 description 1
- 102100038498 Protein maelstrom homolog Human genes 0.000 description 1
- 241000588769 Proteus <enterobacteria> Species 0.000 description 1
- 241000588770 Proteus mirabilis Species 0.000 description 1
- 241000588767 Proteus vulgaris Species 0.000 description 1
- 241000576783 Providencia alcalifaciens Species 0.000 description 1
- 241000588777 Providencia rettgeri Species 0.000 description 1
- 241000588778 Providencia stuartii Species 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 101100084022 Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) lapA gene Proteins 0.000 description 1
- 241000168225 Pseudomonas alcaligenes Species 0.000 description 1
- 241000589540 Pseudomonas fluorescens Species 0.000 description 1
- 241000589776 Pseudomonas putida Species 0.000 description 1
- 102100040001 Putative G antigen family E member 3 Human genes 0.000 description 1
- 102100039359 Putative chondrosarcoma-associated gene 1 protein Human genes 0.000 description 1
- 102100025078 Putative melanoma-associated antigen 5P Human genes 0.000 description 1
- 102100037725 Putative protein SSX6 Human genes 0.000 description 1
- 102100035588 Putative protein SSX9 Human genes 0.000 description 1
- 102100027596 Putative tumor antigen NA88-A Human genes 0.000 description 1
- 102100022578 Putative tyrosine-protein phosphatase TPTE Human genes 0.000 description 1
- AQXXZDYPVDOQEE-MXDQRGINSA-N Pyrantel pamoate Chemical compound CN1CCCN=C1\C=C\C1=CC=CS1.C1=CC=C2C(CC=3C4=CC=CC=C4C=C(C=3O)C(=O)O)=C(O)C(C(O)=O)=CC2=C1 AQXXZDYPVDOQEE-MXDQRGINSA-N 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 102100034188 RING finger protein 17 Human genes 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 description 1
- 101710100968 Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 1
- 102100029165 Receptor-binding cancer antigen expressed on SiSo cells Human genes 0.000 description 1
- 102100021280 Regulator of G-protein signaling 22 Human genes 0.000 description 1
- 101710148116 Regulator of G-protein signaling 22 Proteins 0.000 description 1
- 208000006265 Renal cell carcinoma Diseases 0.000 description 1
- 241000702247 Reoviridae Species 0.000 description 1
- 241000725643 Respiratory syncytial virus Species 0.000 description 1
- 201000000582 Retinoblastoma Diseases 0.000 description 1
- 241000712907 Retroviridae Species 0.000 description 1
- OWPCHSCAPHNHAV-UHFFFAOYSA-N Rhizoxin Natural products C1C(O)C2(C)OC2C=CC(C)C(OC(=O)C2)CC2CC2OC2C(=O)OC1C(C)C(OC)C(C)=CC=CC(C)=CC1=COC(C)=N1 OWPCHSCAPHNHAV-UHFFFAOYSA-N 0.000 description 1
- 102100037754 Rhox homeobox family member 2 Human genes 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 102100032224 Ropporin-1A Human genes 0.000 description 1
- NSFWWJIQIKBZMJ-YKNYLIOZSA-N Roridin A Chemical compound C([C@]12[C@]3(C)[C@H]4C[C@H]1O[C@@H]1C=C(C)CC[C@@]13COC(=O)[C@@H](O)[C@H](C)CCO[C@H](\C=C\C=C/C(=O)O4)[C@H](O)C)O2 NSFWWJIQIKBZMJ-YKNYLIOZSA-N 0.000 description 1
- 241000702670 Rotavirus Species 0.000 description 1
- 241000714474 Rous sarcoma virus Species 0.000 description 1
- 241001533467 Rubulavirus Species 0.000 description 1
- 101150036449 SIRPA gene Proteins 0.000 description 1
- 101150050559 SOAT1 gene Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 101100269260 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) ADH2 gene Proteins 0.000 description 1
- 206010061934 Salivary gland cancer Diseases 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 241001354013 Salmonella enterica subsp. enterica serovar Enteritidis Species 0.000 description 1
- 241001222774 Salmonella enterica subsp. enterica serovar Minnesota Species 0.000 description 1
- 241000531795 Salmonella enterica subsp. enterica serovar Paratyphi A Species 0.000 description 1
- 241000293871 Salmonella enterica subsp. enterica serovar Typhi Species 0.000 description 1
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 1
- 241000192023 Sarcina Species 0.000 description 1
- 102100021466 Sarcoma antigen 1 Human genes 0.000 description 1
- 241000242683 Schistosoma haematobium Species 0.000 description 1
- 241000242677 Schistosoma japonicum Species 0.000 description 1
- 241000242680 Schistosoma mansoni Species 0.000 description 1
- 241001442514 Schistosomatidae Species 0.000 description 1
- 101100071627 Schizosaccharomyces pombe (strain 972 / ATCC 24843) swo1 gene Proteins 0.000 description 1
- 102100037550 Semenogelin-1 Human genes 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 102100031054 Serine protease 55 Human genes 0.000 description 1
- 102100031075 Serine/threonine-protein kinase Chk2 Human genes 0.000 description 1
- 102100026282 Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit alpha isoform Human genes 0.000 description 1
- 241000607720 Serratia Species 0.000 description 1
- 241000607715 Serratia marcescens Species 0.000 description 1
- 241000607768 Shigella Species 0.000 description 1
- 241000607764 Shigella dysenteriae Species 0.000 description 1
- 241000607762 Shigella flexneri Species 0.000 description 1
- 241000607760 Shigella sonnei Species 0.000 description 1
- 102100030403 Signal peptide peptidase-like 2A Human genes 0.000 description 1
- 108050005900 Signal peptide peptidase-like 2a Proteins 0.000 description 1
- 241000713311 Simian immunodeficiency virus Species 0.000 description 1
- 208000000453 Skin Neoplasms Diseases 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 102100032147 Sperm acrosome membrane-associated protein 3 Human genes 0.000 description 1
- 102100030317 Sperm flagellar protein 2 Human genes 0.000 description 1
- 102100022322 Sperm protein associated with the nucleus on the X chromosome C Human genes 0.000 description 1
- 102100031120 Sperm protein associated with the nucleus on the X chromosome N1 Human genes 0.000 description 1
- 102100027689 Sperm protein associated with the nucleus on the X chromosome N2 Human genes 0.000 description 1
- 102100027688 Sperm protein associated with the nucleus on the X chromosome N3 Human genes 0.000 description 1
- 102100027687 Sperm protein associated with the nucleus on the X chromosome N4 Human genes 0.000 description 1
- 102100027686 Sperm protein associated with the nucleus on the X chromosome N5 Human genes 0.000 description 1
- 102100022441 Sperm surface protein Sp17 Human genes 0.000 description 1
- 102100021916 Sperm-associated antigen 1 Human genes 0.000 description 1
- 102100036346 Sperm-associated antigen 17 Human genes 0.000 description 1
- 102100021907 Sperm-associated antigen 4 protein Human genes 0.000 description 1
- 102100021909 Sperm-associated antigen 6 Human genes 0.000 description 1
- 102100021913 Sperm-associated antigen 8 Human genes 0.000 description 1
- 102100036418 Spermatogenesis-associated protein 19, mitochondrial Human genes 0.000 description 1
- 241000710888 St. Louis encephalitis virus Species 0.000 description 1
- 241000191940 Staphylococcus Species 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 241000191963 Staphylococcus epidermidis Species 0.000 description 1
- 241000191984 Staphylococcus haemolyticus Species 0.000 description 1
- 241000192087 Staphylococcus hominis Species 0.000 description 1
- 241000191982 Staphylococcus hyicus Species 0.000 description 1
- 241000191980 Staphylococcus intermedius Species 0.000 description 1
- 241001464905 Staphylococcus saccharolyticus Species 0.000 description 1
- 241001147691 Staphylococcus saprophyticus Species 0.000 description 1
- 241000122973 Stenotrophomonas maltophilia Species 0.000 description 1
- 102100021993 Sterol O-acyltransferase 1 Human genes 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 241000193985 Streptococcus agalactiae Species 0.000 description 1
- 241000193998 Streptococcus pneumoniae Species 0.000 description 1
- 241000193996 Streptococcus pyogenes Species 0.000 description 1
- 241000244177 Strongyloides stercoralis Species 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 102100023532 Synaptic functional regulator FMR1 Human genes 0.000 description 1
- 102100026392 Synaptonemal complex central element protein 1 Human genes 0.000 description 1
- 101710143177 Synaptonemal complex protein 1 Proteins 0.000 description 1
- 102100035721 Syndecan-1 Human genes 0.000 description 1
- 108091008035 T cell costimulatory receptors Proteins 0.000 description 1
- 230000006052 T cell proliferation Effects 0.000 description 1
- BXFOFFBJRFZBQZ-QYWOHJEZSA-N T-2 toxin Chemical compound C([C@@]12[C@]3(C)[C@H](OC(C)=O)[C@@H](O)[C@H]1O[C@H]1[C@]3(COC(C)=O)C[C@@H](C(=C1)C)OC(=O)CC(C)C)O2 BXFOFFBJRFZBQZ-QYWOHJEZSA-N 0.000 description 1
- 229940126547 T-cell immunoglobulin mucin-3 Drugs 0.000 description 1
- 102100024834 T-cell immunoreceptor with Ig and ITIM domains Human genes 0.000 description 1
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 description 1
- 108700012920 TNF Proteins 0.000 description 1
- 102100033082 TNF receptor-associated factor 3 Human genes 0.000 description 1
- 241000244159 Taenia saginata Species 0.000 description 1
- 241000244157 Taenia solium Species 0.000 description 1
- 102100032935 Tektin-5 Human genes 0.000 description 1
- CGMTUJFWROPELF-UHFFFAOYSA-N Tenuazonic acid Natural products CCC(C)C1NC(=O)C(=C(C)/O)C1=O CGMTUJFWROPELF-UHFFFAOYSA-N 0.000 description 1
- 102100038314 Terminal nucleotidyltransferase 5D Human genes 0.000 description 1
- 208000024313 Testicular Neoplasms Diseases 0.000 description 1
- 102100032332 Testicular haploid expressed gene protein Human genes 0.000 description 1
- 206010057644 Testis cancer Diseases 0.000 description 1
- 102100031738 Testis-expressed protein 101 Human genes 0.000 description 1
- 102100035116 Testis-expressed protein 15 Human genes 0.000 description 1
- 102100024994 Testis-specific Y-encoded protein 2 Human genes 0.000 description 1
- 102100024993 Testis-specific Y-encoded protein 3 Human genes 0.000 description 1
- 102100040873 Testis-specific gene 10 protein Human genes 0.000 description 1
- 102100030141 Testis-specific serine/threonine-protein kinase 6 Human genes 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- BHEOSNUKNHRBNM-UHFFFAOYSA-N Tetramethylsqualene Natural products CC(=C)C(C)CCC(=C)C(C)CCC(C)=CCCC=C(C)CCC(C)C(=C)CCC(C)C(C)=C BHEOSNUKNHRBNM-UHFFFAOYSA-N 0.000 description 1
- 210000000068 Th17 cell Anatomy 0.000 description 1
- 210000004241 Th2 cell Anatomy 0.000 description 1
- 108010022394 Threonine synthase Proteins 0.000 description 1
- 102000006601 Thymidine Kinase Human genes 0.000 description 1
- 108020004440 Thymidine kinase Proteins 0.000 description 1
- 208000024770 Thyroid neoplasm Diseases 0.000 description 1
- HJLSLZFTEKNLFI-UHFFFAOYSA-N Tinidazole Chemical compound CCS(=O)(=O)CCN1C(C)=NC=C1[N+]([O-])=O HJLSLZFTEKNLFI-UHFFFAOYSA-N 0.000 description 1
- 102100026159 Tomoregulin-1 Human genes 0.000 description 1
- 102100026160 Tomoregulin-2 Human genes 0.000 description 1
- 241000223997 Toxoplasma gondii Species 0.000 description 1
- 102100038129 Transcription factor Dp family member 3 Human genes 0.000 description 1
- 102100021172 Transcription initiation factor TFIID subunit 7-like Human genes 0.000 description 1
- 102100021393 Transcriptional repressor CTCFL Human genes 0.000 description 1
- 102100026224 Transmembrane and immunoglobulin domain-containing protein 2 Human genes 0.000 description 1
- 102100032469 Transmembrane protease serine 12 Human genes 0.000 description 1
- 102100036709 Transmembrane protein 108 Human genes 0.000 description 1
- 241000242541 Trematoda Species 0.000 description 1
- 241000869417 Trematodes Species 0.000 description 1
- 241000224527 Trichomonas vaginalis Species 0.000 description 1
- 241001489145 Trichuris trichiura Species 0.000 description 1
- UMILHIMHKXVDGH-UHFFFAOYSA-N Triethylene glycol diglycidyl ether Chemical compound C1OC1COCCOCCOCCOCC1CO1 UMILHIMHKXVDGH-UHFFFAOYSA-N 0.000 description 1
- 102100029294 Tubby-related protein 2 Human genes 0.000 description 1
- 102100031935 Tubulin polymerization-promoting protein family member 2 Human genes 0.000 description 1
- 102100040192 Tudor domain-containing protein 1 Human genes 0.000 description 1
- 102100026366 Tudor domain-containing protein 6 Human genes 0.000 description 1
- 108090000138 Tumor necrosis factor ligand superfamily member 15 Proteins 0.000 description 1
- 102100032100 Tumor necrosis factor ligand superfamily member 8 Human genes 0.000 description 1
- 101710165473 Tumor necrosis factor receptor superfamily member 4 Proteins 0.000 description 1
- 102000003425 Tyrosinase Human genes 0.000 description 1
- 108060008724 Tyrosinase Proteins 0.000 description 1
- 102100033019 Tyrosine-protein phosphatase non-receptor type 11 Human genes 0.000 description 1
- 101710116241 Tyrosine-protein phosphatase non-receptor type 11 Proteins 0.000 description 1
- 102100033017 Tyrosine-protein phosphatase non-receptor type 20 Human genes 0.000 description 1
- 102100029948 Tyrosine-protein phosphatase non-receptor type substrate 1 Human genes 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 208000002495 Uterine Neoplasms Diseases 0.000 description 1
- 206010046865 Vaccinia virus infection Diseases 0.000 description 1
- 244000000188 Vaccinium ovalifolium Species 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 108010059993 Vancomycin Proteins 0.000 description 1
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 1
- 102100039037 Vascular endothelial growth factor A Human genes 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 241000607626 Vibrio cholerae Species 0.000 description 1
- 241000607272 Vibrio parahaemolyticus Species 0.000 description 1
- JXLYSJRDGCGARV-WWYNWVTFSA-N Vinblastine Natural products O=C(O[C@H]1[C@](O)(C(=O)OC)[C@@H]2N(C)c3c(cc(c(OC)c3)[C@]3(C(=O)OC)c4[nH]c5c(c4CCN4C[C@](O)(CC)C[C@H](C3)C4)cccc5)[C@@]32[C@H]2[C@@]1(CC)C=CCN2CC3)C JXLYSJRDGCGARV-WWYNWVTFSA-N 0.000 description 1
- 108700005077 Viral Genes Proteins 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 206010047741 Vulval cancer Diseases 0.000 description 1
- 208000033559 Waldenström macroglobulinemia Diseases 0.000 description 1
- 241000710886 West Nile virus Species 0.000 description 1
- 241000244005 Wuchereria bancrofti Species 0.000 description 1
- 102100039491 X antigen family member 3 Human genes 0.000 description 1
- 102100039494 X antigen family member 5 Human genes 0.000 description 1
- 241000710772 Yellow fever virus Species 0.000 description 1
- 241000607447 Yersinia enterocolitica Species 0.000 description 1
- 241000607481 Yersinia intermedia Species 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 241000607477 Yersinia pseudotuberculosis Species 0.000 description 1
- 102100040814 Zinc finger protein 165 Human genes 0.000 description 1
- UZQJVUCHXGYFLQ-AYDHOLPZSA-N [(2s,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-4-[(2r,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-3,5-dihydroxy-6-(hydroxymethyl)-4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-3,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3,5-dihydroxy-6-(hy Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O)O[C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O)O[C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2[C@@]1(C=O)C)C)(C)CC(O)[C@]1(CCC(CC14)(C)C)C(=O)O[C@H]1[C@@H]([C@@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O[C@H]4[C@@H]([C@@H](O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O)[C@H](O)[C@@H](CO)O4)O)[C@H](O)[C@@H](CO)O3)O)[C@H](O)[C@@H](CO)O2)O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UZQJVUCHXGYFLQ-AYDHOLPZSA-N 0.000 description 1
- SPJCRMJCFSJKDE-ZWBUGVOYSA-N [(3s,8s,9s,10r,13r,14s,17r)-10,13-dimethyl-17-[(2r)-6-methylheptan-2-yl]-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1h-cyclopenta[a]phenanthren-3-yl] 2-[4-[bis(2-chloroethyl)amino]phenyl]acetate Chemical compound O([C@@H]1CC2=CC[C@H]3[C@@H]4CC[C@@H]([C@]4(CC[C@@H]3[C@@]2(C)CC1)C)[C@H](C)CCCC(C)C)C(=O)CC1=CC=C(N(CCCl)CCCl)C=C1 SPJCRMJCFSJKDE-ZWBUGVOYSA-N 0.000 description 1
- IFJUINDAXYAPTO-UUBSBJJBSA-N [(8r,9s,13s,14s,17s)-17-[2-[4-[4-[bis(2-chloroethyl)amino]phenyl]butanoyloxy]acetyl]oxy-13-methyl-6,7,8,9,11,12,14,15,16,17-decahydrocyclopenta[a]phenanthren-3-yl] benzoate Chemical compound C([C@@H]1[C@@H](C2=CC=3)CC[C@]4([C@H]1CC[C@@H]4OC(=O)COC(=O)CCCC=1C=CC(=CC=1)N(CCCl)CCCl)C)CC2=CC=3OC(=O)C1=CC=CC=C1 IFJUINDAXYAPTO-UUBSBJJBSA-N 0.000 description 1
- XZSRRNFBEIOBDA-CFNBKWCHSA-N [2-[(2s,4s)-4-[(2r,4s,5s,6s)-4-amino-5-hydroxy-6-methyloxan-2-yl]oxy-2,5,12-trihydroxy-7-methoxy-6,11-dioxo-3,4-dihydro-1h-tetracen-2-yl]-2-oxoethyl] 2,2-diethoxyacetate Chemical compound O([C@H]1C[C@](CC2=C(O)C=3C(=O)C4=CC=CC(OC)=C4C(=O)C=3C(O)=C21)(O)C(=O)COC(=O)C(OCC)OCC)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 XZSRRNFBEIOBDA-CFNBKWCHSA-N 0.000 description 1
- 241000606834 [Haemophilus] ducreyi Species 0.000 description 1
- MCGSCOLBFJQGHM-SCZZXKLOSA-N abacavir Chemical compound C=12N=CN([C@H]3C=C[C@@H](CO)C3)C2=NC(N)=NC=1NC1CC1 MCGSCOLBFJQGHM-SCZZXKLOSA-N 0.000 description 1
- 229960004748 abacavir Drugs 0.000 description 1
- 229940028652 abraxane Drugs 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- ZOZKYEHVNDEUCO-XUTVFYLZSA-N aceglatone Chemical compound O1C(=O)[C@H](OC(C)=O)[C@@H]2OC(=O)[C@@H](OC(=O)C)[C@@H]21 ZOZKYEHVNDEUCO-XUTVFYLZSA-N 0.000 description 1
- 229950002684 aceglatone Drugs 0.000 description 1
- YRKCREAYFQTBPV-UHFFFAOYSA-N acetylacetone Chemical compound CC(=O)CC(C)=O YRKCREAYFQTBPV-UHFFFAOYSA-N 0.000 description 1
- 229960004150 aciclovir Drugs 0.000 description 1
- MKUXAQIIEYXACX-UHFFFAOYSA-N aciclovir Chemical compound N1C(N)=NC(=O)C2=C1N(COCCO)C=N2 MKUXAQIIEYXACX-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 229930183665 actinomycin Natural products 0.000 description 1
- RJURFGZVJUQBHK-IIXSONLDSA-N actinomycin D Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)N[C@@H]4C(=O)N[C@@H](C(N5CCC[C@H]5C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-IIXSONLDSA-N 0.000 description 1
- 229960001997 adefovir Drugs 0.000 description 1
- WOZSCQDILHKSGG-UHFFFAOYSA-N adefovir depivoxil Chemical compound N1=CN=C2N(CCOCP(=O)(OCOC(=O)C(C)(C)C)OCOC(=O)C(C)(C)C)C=NC2=C1N WOZSCQDILHKSGG-UHFFFAOYSA-N 0.000 description 1
- 208000009956 adenocarcinoma Diseases 0.000 description 1
- 230000000240 adjuvant effect Effects 0.000 description 1
- 229950004955 adozelesin Drugs 0.000 description 1
- BYRVKDUQDLJUBX-JJCDCTGGSA-N adozelesin Chemical compound C1=CC=C2OC(C(=O)NC=3C=C4C=C(NC4=CC=3)C(=O)N3C[C@H]4C[C@]44C5=C(C(C=C43)=O)NC=C5C)=CC2=C1 BYRVKDUQDLJUBX-JJCDCTGGSA-N 0.000 description 1
- 229940009456 adriamycin Drugs 0.000 description 1
- 208000037844 advanced solid tumor Diseases 0.000 description 1
- 241001148470 aerobic bacillus Species 0.000 description 1
- 230000001270 agonistic effect Effects 0.000 description 1
- HXHWSAZORRCQMX-UHFFFAOYSA-N albendazole Chemical compound CCCSC1=CC=C2NC(NC(=O)OC)=NC2=C1 HXHWSAZORRCQMX-UHFFFAOYSA-N 0.000 description 1
- 229960002669 albendazole Drugs 0.000 description 1
- 229940045714 alkyl sulfonate alkylating agent Drugs 0.000 description 1
- 150000008052 alkyl sulfonates Chemical class 0.000 description 1
- 229940100198 alkylating agent Drugs 0.000 description 1
- 239000002168 alkylating agent Substances 0.000 description 1
- SHGAZHPCJJPHSC-YCNIQYBTSA-N all-trans-retinoic acid Chemical compound OC(=O)\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-YCNIQYBTSA-N 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- 108010050122 alpha 1-Antitrypsin Proteins 0.000 description 1
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 1
- 229960000473 altretamine Drugs 0.000 description 1
- 229940037003 alum Drugs 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 229960004050 aminobenzoic acid Drugs 0.000 description 1
- 229960002684 aminocaproic acid Drugs 0.000 description 1
- 229940126575 aminoglycoside Drugs 0.000 description 1
- 229960002749 aminolevulinic acid Drugs 0.000 description 1
- 229960003896 aminopterin Drugs 0.000 description 1
- 229960003022 amoxicillin Drugs 0.000 description 1
- LSQZJLSUYDQPKJ-NJBDSQKTSA-N amoxicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=C(O)C=C1 LSQZJLSUYDQPKJ-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- YMARZQAQMVYCKC-OEMFJLHTSA-N amprenavir Chemical compound C([C@@H]([C@H](O)CN(CC(C)C)S(=O)(=O)C=1C=CC(N)=CC=1)NC(=O)O[C@@H]1COCC1)C1=CC=CC=C1 YMARZQAQMVYCKC-OEMFJLHTSA-N 0.000 description 1
- 229960001830 amprenavir Drugs 0.000 description 1
- 229960001220 amsacrine Drugs 0.000 description 1
- XCPGHVQEEXUHNC-UHFFFAOYSA-N amsacrine Chemical compound COC1=CC(NS(C)(=O)=O)=CC=C1NC1=C(C=CC=C2)C2=NC2=CC=CC=C12 XCPGHVQEEXUHNC-UHFFFAOYSA-N 0.000 description 1
- BBDAGFIXKZCXAH-CCXZUQQUSA-N ancitabine Chemical compound N=C1C=CN2[C@@H]3O[C@H](CO)[C@@H](O)[C@@H]3OC2=N1 BBDAGFIXKZCXAH-CCXZUQQUSA-N 0.000 description 1
- 229950000242 ancitabine Drugs 0.000 description 1
- 239000003098 androgen Substances 0.000 description 1
- 229940030486 androgens Drugs 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000003466 anti-cipated effect Effects 0.000 description 1
- 230000003388 anti-hormonal effect Effects 0.000 description 1
- 230000006023 anti-tumor response Effects 0.000 description 1
- 238000011319 anticancer therapy Methods 0.000 description 1
- 239000003430 antimalarial agent Substances 0.000 description 1
- 229940045687 antimetabolites folic acid analogs Drugs 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 150000008209 arabinosides Chemical class 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 125000000637 arginyl group Chemical class N[C@@H](CCCNC(N)=N)C(=O)* 0.000 description 1
- 229940081238 artemether / lumefantrine Drugs 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000001174 ascending effect Effects 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- AXRYRYVKAWYZBR-GASGPIRDSA-N atazanavir Chemical compound C([C@H](NC(=O)[C@@H](NC(=O)OC)C(C)(C)C)[C@@H](O)CN(CC=1C=CC(=CC=1)C=1N=CC=CC=1)NC(=O)[C@@H](NC(=O)OC)C(C)(C)C)C1=CC=CC=C1 AXRYRYVKAWYZBR-GASGPIRDSA-N 0.000 description 1
- 229960003277 atazanavir Drugs 0.000 description 1
- 229960003852 atezolizumab Drugs 0.000 description 1
- 229940114027 atovaquone / proguanil Drugs 0.000 description 1
- 229940062316 avelox Drugs 0.000 description 1
- 229960002756 azacitidine Drugs 0.000 description 1
- VSRXQHXAPYXROS-UHFFFAOYSA-N azanide;cyclobutane-1,1-dicarboxylic acid;platinum(2+) Chemical compound [NH2-].[NH2-].[Pt+2].OC(=O)C1(C(O)=O)CCC1 VSRXQHXAPYXROS-UHFFFAOYSA-N 0.000 description 1
- 229950011321 azaserine Drugs 0.000 description 1
- 150000001541 aziridines Chemical class 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 208000003373 basosquamous carcinoma Diseases 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 201000009036 biliary tract cancer Diseases 0.000 description 1
- 208000020790 biliary tract neoplasm Diseases 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 238000007622 bioinformatic analysis Methods 0.000 description 1
- 238000001815 biotherapy Methods 0.000 description 1
- 229950008548 bisantrene Drugs 0.000 description 1
- 150000004663 bisphosphonates Chemical class 0.000 description 1
- 229950006844 bizelesin Drugs 0.000 description 1
- OYVAGSVQBOHSSS-UAPAGMARSA-O bleomycin A2 Chemical class N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCC[S+](C)C)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C OYVAGSVQBOHSSS-UAPAGMARSA-O 0.000 description 1
- 229920001400 block copolymer Polymers 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 201000000220 brain stem cancer Diseases 0.000 description 1
- 201000008275 breast carcinoma Diseases 0.000 description 1
- 229960005520 bryostatin Drugs 0.000 description 1
- MJQUEDHRCUIRLF-TVIXENOKSA-N bryostatin 1 Chemical compound C([C@@H]1CC(/[C@@H]([C@@](C(C)(C)/C=C/2)(O)O1)OC(=O)/C=C/C=C/CCC)=C\C(=O)OC)[C@H]([C@@H](C)O)OC(=O)C[C@H](O)C[C@@H](O1)C[C@H](OC(C)=O)C(C)(C)[C@]1(O)C[C@@H]1C\C(=C\C(=O)OC)C[C@H]\2O1 MJQUEDHRCUIRLF-TVIXENOKSA-N 0.000 description 1
- MUIWQCKLQMOUAT-AKUNNTHJSA-N bryostatin 20 Natural products COC(=O)C=C1C[C@@]2(C)C[C@]3(O)O[C@](C)(C[C@@H](O)CC(=O)O[C@](C)(C[C@@]4(C)O[C@](O)(CC5=CC(=O)O[C@]45C)C(C)(C)C=C[C@@](C)(C1)O2)[C@@H](C)O)C[C@H](OC(=O)C(C)(C)C)C3(C)C MUIWQCKLQMOUAT-AKUNNTHJSA-N 0.000 description 1
- MBABCNBNDNGODA-LUVUIASKSA-N bullatacin Chemical compound O1[C@@H]([C@@H](O)CCCCCCCCCC)CC[C@@H]1[C@@H]1O[C@@H]([C@H](O)CCCCCCCCCC[C@@H](O)CC=2C(O[C@@H](C)C=2)=O)CC1 MBABCNBNDNGODA-LUVUIASKSA-N 0.000 description 1
- 229960002092 busulfan Drugs 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 108700002839 cactinomycin Proteins 0.000 description 1
- 229950009908 cactinomycin Drugs 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 235000011132 calcium sulphate Nutrition 0.000 description 1
- 238000004422 calculation algorithm Methods 0.000 description 1
- IVFYLRMMHVYGJH-PVPPCFLZSA-N calusterone Chemical compound C1C[C@]2(C)[C@](O)(C)CC[C@H]2[C@@H]2[C@@H](C)CC3=CC(=O)CC[C@]3(C)[C@H]21 IVFYLRMMHVYGJH-PVPPCFLZSA-N 0.000 description 1
- 229950009823 calusterone Drugs 0.000 description 1
- 229940088954 camptosar Drugs 0.000 description 1
- 229940127093 camptothecin Drugs 0.000 description 1
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical compound C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 description 1
- 229960004117 capecitabine Drugs 0.000 description 1
- YZBQHRLRFGPBSL-RXMQYKEDSA-N carbapenem Chemical compound C1C=CN2C(=O)C[C@H]21 YZBQHRLRFGPBSL-RXMQYKEDSA-N 0.000 description 1
- 229960003669 carbenicillin Drugs 0.000 description 1
- FPPNZSSZRUTDAP-UWFZAAFLSA-N carbenicillin Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)C(C(O)=O)C1=CC=CC=C1 FPPNZSSZRUTDAP-UWFZAAFLSA-N 0.000 description 1
- 239000011545 carbonate/bicarbonate buffer Substances 0.000 description 1
- 229960004562 carboplatin Drugs 0.000 description 1
- 229960002115 carboquone Drugs 0.000 description 1
- 229960003261 carmofur Drugs 0.000 description 1
- 229960005243 carmustine Drugs 0.000 description 1
- 229950007509 carzelesin Drugs 0.000 description 1
- BBZDXMBRAFTCAA-AREMUKBSSA-N carzelesin Chemical compound C1=2NC=C(C)C=2C([C@H](CCl)CN2C(=O)C=3NC4=CC=C(C=C4C=3)NC(=O)C3=CC4=CC=C(C=C4O3)N(CC)CC)=C2C=C1OC(=O)NC1=CC=CC=C1 BBZDXMBRAFTCAA-AREMUKBSSA-N 0.000 description 1
- 108010047060 carzinophilin Proteins 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 210000004970 cd4 cell Anatomy 0.000 description 1
- 229960005361 cefaclor Drugs 0.000 description 1
- QYIYFLOTGYLRGG-GPCCPHFNSA-N cefaclor Chemical compound C1([C@H](C(=O)N[C@@H]2C(N3C(=C(Cl)CS[C@@H]32)C(O)=O)=O)N)=CC=CC=C1 QYIYFLOTGYLRGG-GPCCPHFNSA-N 0.000 description 1
- 229960004841 cefadroxil Drugs 0.000 description 1
- NBFNMSULHIODTC-CYJZLJNKSA-N cefadroxil monohydrate Chemical compound O.C1([C@@H](N)C(=O)N[C@H]2[C@@H]3N(C2=O)C(=C(CS3)C)C(O)=O)=CC=C(O)C=C1 NBFNMSULHIODTC-CYJZLJNKSA-N 0.000 description 1
- 229960000603 cefalotin Drugs 0.000 description 1
- 229960003012 cefamandole Drugs 0.000 description 1
- OLVCFLKTBJRLHI-AXAPSJFSSA-N cefamandole Chemical compound CN1N=NN=C1SCC1=C(C(O)=O)N2C(=O)[C@@H](NC(=O)[C@H](O)C=3C=CC=CC=3)[C@H]2SC1 OLVCFLKTBJRLHI-AXAPSJFSSA-N 0.000 description 1
- 229960001139 cefazolin Drugs 0.000 description 1
- MLYYVTUWGNIJIB-BXKDBHETSA-N cefazolin Chemical compound S1C(C)=NN=C1SCC1=C(C(O)=O)N2C(=O)[C@@H](NC(=O)CN3N=NN=C3)[C@H]2SC1 MLYYVTUWGNIJIB-BXKDBHETSA-N 0.000 description 1
- 229960002682 cefoxitin Drugs 0.000 description 1
- 229960002580 cefprozil Drugs 0.000 description 1
- VOAZJEPQLGBXGO-SDAWRPRTSA-N ceftobiprole Chemical compound S1C(N)=NC(C(=N\O)\C(=O)N[C@@H]2C(N3C(=C(\C=C/4C(N([C@H]5CNCC5)CC\4)=O)CS[C@@H]32)C(O)=O)=O)=N1 VOAZJEPQLGBXGO-SDAWRPRTSA-N 0.000 description 1
- 229950004259 ceftobiprole Drugs 0.000 description 1
- 229960001668 cefuroxime Drugs 0.000 description 1
- JFPVXVDWJQMJEE-IZRZKJBUSA-N cefuroxime Chemical compound N([C@@H]1C(N2C(=C(COC(N)=O)CS[C@@H]21)C(O)=O)=O)C(=O)\C(=N/OC)C1=CC=CO1 JFPVXVDWJQMJEE-IZRZKJBUSA-N 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000009134 cell regulation Effects 0.000 description 1
- 238000002659 cell therapy Methods 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 210000004520 cell wall skeleton Anatomy 0.000 description 1
- 230000036755 cellular response Effects 0.000 description 1
- 210000002230 centromere Anatomy 0.000 description 1
- 229940106164 cephalexin Drugs 0.000 description 1
- ZAIPMKNFIOOWCQ-UEKVPHQBSA-N cephalexin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@@H]3N(C2=O)C(=C(CS3)C)C(O)=O)=CC=CC=C1 ZAIPMKNFIOOWCQ-UEKVPHQBSA-N 0.000 description 1
- 229940124587 cephalosporin Drugs 0.000 description 1
- 150000001780 cephalosporins Chemical class 0.000 description 1
- VUFGUVLLDPOSBC-XRZFDKQNSA-M cephalothin sodium Chemical compound [Na+].N([C@H]1[C@@H]2N(C1=O)C(=C(CS2)COC(=O)C)C([O-])=O)C(=O)CC1=CC=CS1 VUFGUVLLDPOSBC-XRZFDKQNSA-M 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 208000019065 cervical carcinoma Diseases 0.000 description 1
- 229960004630 chlorambucil Drugs 0.000 description 1
- JCKYGMPEJWAADB-UHFFFAOYSA-N chlorambucil Chemical compound OC(=O)CCCC1=CC=C(N(CCCl)CCCl)C=C1 JCKYGMPEJWAADB-UHFFFAOYSA-N 0.000 description 1
- 229950008249 chlornaphazine Drugs 0.000 description 1
- 229960003677 chloroquine Drugs 0.000 description 1
- WHTVZRBIWZFKQO-UHFFFAOYSA-N chloroquine Natural products ClC1=CC=C2C(NC(C)CCCN(CC)CC)=CC=NC2=C1 WHTVZRBIWZFKQO-UHFFFAOYSA-N 0.000 description 1
- 229960001480 chlorozotocin Drugs 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 229960000724 cidofovir Drugs 0.000 description 1
- LOUPRKONTZGTKE-UHFFFAOYSA-N cinchonine Natural products C1C(C(C2)C=C)CCN2C1C(O)C1=CC=NC2=CC=C(OC)C=C21 LOUPRKONTZGTKE-UHFFFAOYSA-N 0.000 description 1
- 229940088516 cipro Drugs 0.000 description 1
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 description 1
- 229960004316 cisplatin Drugs 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 229960002173 citrulline Drugs 0.000 description 1
- 235000013477 citrulline Nutrition 0.000 description 1
- ACSIXWWBWUQEHA-UHFFFAOYSA-N clodronic acid Chemical compound OP(O)(=O)C(Cl)(Cl)P(O)(O)=O ACSIXWWBWUQEHA-UHFFFAOYSA-N 0.000 description 1
- 229960002286 clodronic acid Drugs 0.000 description 1
- 238000012411 cloning technique Methods 0.000 description 1
- 239000013599 cloning vector Substances 0.000 description 1
- 238000011260 co-administration Methods 0.000 description 1
- ZVAQGQOEHFIYMQ-PRLJFWCFSA-N co-artemether Chemical compound C1C[C@H]2[C@H](C)CC[C@H]3[C@@H](C)[C@@H](OC)O[C@H]4[C@]32OOC1(C)O4.C12=CC(Cl)=CC=C2C=2C(C(O)CN(CCCC)CCCC)=CC(Cl)=CC=2\C1=C/C1=CC=C(Cl)C=C1 ZVAQGQOEHFIYMQ-PRLJFWCFSA-N 0.000 description 1
- 238000000975 co-precipitation Methods 0.000 description 1
- 238000011220 combination immunotherapy Methods 0.000 description 1
- LGZKGOGODCLQHG-UHFFFAOYSA-N combretastatin Natural products C1=C(O)C(OC)=CC=C1CC(O)C1=CC(OC)=C(OC)C(OC)=C1 LGZKGOGODCLQHG-UHFFFAOYSA-N 0.000 description 1
- 238000004590 computer program Methods 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 201000010918 connective tissue cancer Diseases 0.000 description 1
- 108010015408 connexin 37 Proteins 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 231100001159 controllable toxicity Toxicity 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000003246 corticosteroid Substances 0.000 description 1
- 229960001334 corticosteroids Drugs 0.000 description 1
- 230000004940 costimulation Effects 0.000 description 1
- 108010089438 cryptophycin 1 Proteins 0.000 description 1
- PSNOPSMXOBPNNV-VVCTWANISA-N cryptophycin 1 Chemical compound C1=C(Cl)C(OC)=CC=C1C[C@@H]1C(=O)NC[C@@H](C)C(=O)O[C@@H](CC(C)C)C(=O)O[C@H]([C@H](C)[C@@H]2[C@H](O2)C=2C=CC=CC=2)C/C=C/C(=O)N1 PSNOPSMXOBPNNV-VVCTWANISA-N 0.000 description 1
- 108010090203 cryptophycin 8 Proteins 0.000 description 1
- PSNOPSMXOBPNNV-UHFFFAOYSA-N cryptophycin-327 Natural products C1=C(Cl)C(OC)=CC=C1CC1C(=O)NCC(C)C(=O)OC(CC(C)C)C(=O)OC(C(C)C2C(O2)C=2C=CC=CC=2)CC=CC(=O)N1 PSNOPSMXOBPNNV-UHFFFAOYSA-N 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000013211 curve analysis Methods 0.000 description 1
- 108010048032 cyclophilin B Proteins 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- 229960000684 cytarabine Drugs 0.000 description 1
- 229960003901 dacarbazine Drugs 0.000 description 1
- 229960000640 dactinomycin Drugs 0.000 description 1
- CJBJHOAVZSMMDJ-HEXNFIEUSA-N darunavir Chemical compound C([C@@H]([C@H](O)CN(CC(C)C)S(=O)(=O)C=1C=CC(N)=CC=1)NC(=O)O[C@@H]1[C@@H]2CCO[C@@H]2OC1)C1=CC=CC=C1 CJBJHOAVZSMMDJ-HEXNFIEUSA-N 0.000 description 1
- 229960005107 darunavir Drugs 0.000 description 1
- 229960000975 daunorubicin Drugs 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 229960005319 delavirdine Drugs 0.000 description 1
- 108010031971 delta catenin Proteins 0.000 description 1
- 229960005052 demecolcine Drugs 0.000 description 1
- 229950003913 detorubicin Drugs 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- WVYXNIXAMZOZFK-UHFFFAOYSA-N diaziquone Chemical compound O=C1C(NC(=O)OCC)=C(N2CC2)C(=O)C(NC(=O)OCC)=C1N1CC1 WVYXNIXAMZOZFK-UHFFFAOYSA-N 0.000 description 1
- 229950002389 diaziquone Drugs 0.000 description 1
- 229960001585 dicloxacillin Drugs 0.000 description 1
- YFAGHNZHGGCZAX-JKIFEVAISA-N dicloxacillin Chemical compound N([C@@H]1C(N2[C@H](C(C)(C)S[C@@H]21)C(O)=O)=O)C(=O)C1=C(C)ON=C1C1=C(Cl)C=CC=C1Cl YFAGHNZHGGCZAX-JKIFEVAISA-N 0.000 description 1
- 229960002656 didanosine Drugs 0.000 description 1
- 102000004419 dihydrofolate reductase Human genes 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000006334 disulfide bridging Effects 0.000 description 1
- VSJKWCGYPAHWDS-UHFFFAOYSA-N dl-camptothecin Natural products C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)C5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-UHFFFAOYSA-N 0.000 description 1
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 description 1
- 239000003534 dna topoisomerase inhibitor Substances 0.000 description 1
- 101150115114 dnaJ gene Proteins 0.000 description 1
- 101150052825 dnaK gene Proteins 0.000 description 1
- 229960000735 docosanol Drugs 0.000 description 1
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N dodecahydrosqualene Natural products CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 description 1
- AMRJKAQTDDKMCE-UHFFFAOYSA-N dolastatin Chemical compound CC(C)C(N(C)C)C(=O)NC(C(C)C)C(=O)N(C)C(C(C)C)C(OC)CC(=O)N1CCCC1C(OC)C(C)C(=O)NC(C=1SC=CN=1)CC1=CC=CC=C1 AMRJKAQTDDKMCE-UHFFFAOYSA-N 0.000 description 1
- 229930188854 dolastatin Natural products 0.000 description 1
- 229960000895 doripenem Drugs 0.000 description 1
- AVAACINZEOAHHE-VFZPANTDSA-N doripenem Chemical compound C=1([C@H](C)[C@@H]2[C@H](C(N2C=1C(O)=O)=O)[C@H](O)C)S[C@@H]1CN[C@H](CNS(N)(=O)=O)C1 AVAACINZEOAHHE-VFZPANTDSA-N 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- ZWAOHEXOSAUJHY-ZIYNGMLESA-N doxifluridine Chemical compound O[C@@H]1[C@H](O)[C@@H](C)O[C@H]1N1C(=O)NC(=O)C(F)=C1 ZWAOHEXOSAUJHY-ZIYNGMLESA-N 0.000 description 1
- 229950005454 doxifluridine Drugs 0.000 description 1
- 229960004679 doxorubicin Drugs 0.000 description 1
- NOTIQUSPUUHHEH-UXOVVSIBSA-N dromostanolone propionate Chemical compound C([C@@H]1CC2)C(=O)[C@H](C)C[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H](OC(=O)CC)[C@@]2(C)CC1 NOTIQUSPUUHHEH-UXOVVSIBSA-N 0.000 description 1
- 229950004683 drostanolone propionate Drugs 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 229960005501 duocarmycin Drugs 0.000 description 1
- VQNATVDKACXKTF-XELLLNAOSA-N duocarmycin Chemical compound COC1=C(OC)C(OC)=C2NC(C(=O)N3C4=CC(=O)C5=C([C@@]64C[C@@H]6C3)C=C(N5)C(=O)OC)=CC2=C1 VQNATVDKACXKTF-XELLLNAOSA-N 0.000 description 1
- 229930184221 duocarmycin Natural products 0.000 description 1
- AFMYMMXSQGUCBK-AKMKHHNQSA-N dynemicin a Chemical compound C1#C\C=C/C#C[C@@H]2NC(C=3C(=O)C4=C(O)C=CC(O)=C4C(=O)C=3C(O)=C3)=C3[C@@]34O[C@]32[C@@H](C)C(C(O)=O)=C(OC)[C@H]41 AFMYMMXSQGUCBK-AKMKHHNQSA-N 0.000 description 1
- 230000008482 dysregulation Effects 0.000 description 1
- FSIRXIHZBIXHKT-MHTVFEQDSA-N edatrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CC(CC)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FSIRXIHZBIXHKT-MHTVFEQDSA-N 0.000 description 1
- 229950006700 edatrexate Drugs 0.000 description 1
- XPOQHMRABVBWPR-ZDUSSCGKSA-N efavirenz Chemical compound C([C@]1(C2=CC(Cl)=CC=C2NC(=O)O1)C(F)(F)F)#CC1CC1 XPOQHMRABVBWPR-ZDUSSCGKSA-N 0.000 description 1
- 229960003804 efavirenz Drugs 0.000 description 1
- 210000003162 effector t lymphocyte Anatomy 0.000 description 1
- VLCYCQAOQCDTCN-UHFFFAOYSA-N eflornithine Chemical compound NCCCC(N)(C(F)F)C(O)=O VLCYCQAOQCDTCN-UHFFFAOYSA-N 0.000 description 1
- 238000002565 electrocardiography Methods 0.000 description 1
- XOPYFXBZMVTEJF-UHFFFAOYSA-N eleutherobin Natural products C1=CC2(OC)OC1(C)C(OC(=O)C=CC=1N=CN(C)C=1)CC(C(=CCC1C(C)C)C)C1C=C2COC1OCC(O)C(O)C1OC(C)=O XOPYFXBZMVTEJF-UHFFFAOYSA-N 0.000 description 1
- XOPYFXBZMVTEJF-PDACKIITSA-N eleutherobin Chemical compound C(/[C@H]1[C@H](C(=CC[C@@H]1C(C)C)C)C[C@@H]([C@@]1(C)O[C@@]2(C=C1)OC)OC(=O)\C=C\C=1N=CN(C)C=1)=C2\CO[C@@H]1OC[C@@H](O)[C@@H](O)[C@@H]1OC(C)=O XOPYFXBZMVTEJF-PDACKIITSA-N 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 229950000549 elliptinium acetate Drugs 0.000 description 1
- JUZYLCPPVHEVSV-LJQANCHMSA-N elvitegravir Chemical compound COC1=CC=2N([C@H](CO)C(C)C)C=C(C(O)=O)C(=O)C=2C=C1CC1=CC=CC(Cl)=C1F JUZYLCPPVHEVSV-LJQANCHMSA-N 0.000 description 1
- 229960003586 elvitegravir Drugs 0.000 description 1
- 229960000366 emtricitabine Drugs 0.000 description 1
- 230000002121 endocytic effect Effects 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 229960002062 enfuvirtide Drugs 0.000 description 1
- PEASPLKKXBYDKL-FXEVSJAOSA-N enfuvirtide Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(C)=O)[C@@H](C)O)[C@@H](C)CC)C1=CN=CN1 PEASPLKKXBYDKL-FXEVSJAOSA-N 0.000 description 1
- JOZGNYDSEBIJDH-UHFFFAOYSA-N eniluracil Chemical compound O=C1NC=C(C#C)C(=O)N1 JOZGNYDSEBIJDH-UHFFFAOYSA-N 0.000 description 1
- 229950010213 eniluracil Drugs 0.000 description 1
- 229950011487 enocitabine Drugs 0.000 description 1
- 229940092559 enterobacter aerogenes Drugs 0.000 description 1
- 229940032049 enterococcus faecalis Drugs 0.000 description 1
- 210000003979 eosinophil Anatomy 0.000 description 1
- 102000052116 epidermal growth factor receptor activity proteins Human genes 0.000 description 1
- 108700015053 epidermal growth factor receptor activity proteins Proteins 0.000 description 1
- 229960001904 epirubicin Drugs 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 229950002973 epitiostanol Drugs 0.000 description 1
- 229930013356 epothilone Natural products 0.000 description 1
- 150000003883 epothilone derivatives Chemical class 0.000 description 1
- AAKJLRGGTJKAMG-UHFFFAOYSA-N erlotinib Chemical compound C=12C=C(OCCOC)C(OCCOC)=CC2=NC=NC=1NC1=CC=CC(C#C)=C1 AAKJLRGGTJKAMG-UHFFFAOYSA-N 0.000 description 1
- 229960002770 ertapenem Drugs 0.000 description 1
- 201000004101 esophageal cancer Diseases 0.000 description 1
- 229950002017 esorubicin Drugs 0.000 description 1
- ITSGNOIFAJAQHJ-BMFNZSJVSA-N esorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)C[C@H](C)O1 ITSGNOIFAJAQHJ-BMFNZSJVSA-N 0.000 description 1
- LJQQFQHBKUKHIS-WJHRIEJJSA-N esperamicin Chemical compound O1CC(NC(C)C)C(OC)CC1OC1C(O)C(NOC2OC(C)C(SC)C(O)C2)C(C)OC1OC1C(\C2=C/CSSSC)=C(NC(=O)OC)C(=O)C(OC3OC(C)C(O)C(OC(=O)C=4C(=CC(OC)=C(OC)C=4)NC(=O)C(=C)OC)C3)C2(O)C#C\C=C/C#C1 LJQQFQHBKUKHIS-WJHRIEJJSA-N 0.000 description 1
- 229960001842 estramustine Drugs 0.000 description 1
- FRPJXPJMRWBBIH-RBRWEJTLSA-N estramustine Chemical compound ClCCN(CCCl)C(=O)OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 FRPJXPJMRWBBIH-RBRWEJTLSA-N 0.000 description 1
- QSRLNKCNOLVZIR-KRWDZBQOSA-N ethyl (2s)-2-[[2-[4-[bis(2-chloroethyl)amino]phenyl]acetyl]amino]-4-methylsulfanylbutanoate Chemical compound CCOC(=O)[C@H](CCSC)NC(=O)CC1=CC=C(N(CCCl)CCCl)C=C1 QSRLNKCNOLVZIR-KRWDZBQOSA-N 0.000 description 1
- 229960005237 etoglucid Drugs 0.000 description 1
- VJJPUSNTGOMMGY-MRVIYFEKSA-N etoposide Chemical compound COC1=C(O)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@@H](O[C@H]3[C@@H]([C@@H](O)[C@@H]4O[C@H](C)OC[C@H]4O3)O)[C@@H]3[C@@H]2C(OC3)=O)=C1 VJJPUSNTGOMMGY-MRVIYFEKSA-N 0.000 description 1
- PYGWGZALEOIKDF-UHFFFAOYSA-N etravirine Chemical compound CC1=CC(C#N)=CC(C)=C1OC1=NC(NC=2C=CC(=CC=2)C#N)=NC(N)=C1Br PYGWGZALEOIKDF-UHFFFAOYSA-N 0.000 description 1
- 229960002049 etravirine Drugs 0.000 description 1
- 208000021045 exocrine pancreatic carcinoma Diseases 0.000 description 1
- 239000003777 experimental drug Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 208000024519 eye neoplasm Diseases 0.000 description 1
- 229960004396 famciclovir Drugs 0.000 description 1
- GGXKWVWZWMLJEH-UHFFFAOYSA-N famcyclovir Chemical compound N1=C(N)N=C2N(CCC(COC(=O)C)COC(C)=O)C=NC2=C1 GGXKWVWZWMLJEH-UHFFFAOYSA-N 0.000 description 1
- 208000006275 fascioliasis Diseases 0.000 description 1
- 206010016235 fasciolopsiasis Diseases 0.000 description 1
- 206010016256 fatigue Diseases 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 229940072686 floxin Drugs 0.000 description 1
- 229960000961 floxuridine Drugs 0.000 description 1
- ODKNJVUHOIMIIZ-RRKCRQDMSA-N floxuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(F)=C1 ODKNJVUHOIMIIZ-RRKCRQDMSA-N 0.000 description 1
- 229960000390 fludarabine Drugs 0.000 description 1
- GIUYCYHIANZCFB-FJFJXFQQSA-N fludarabine phosphate Chemical compound C1=NC=2C(N)=NC(F)=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@@H]1O GIUYCYHIANZCFB-FJFJXFQQSA-N 0.000 description 1
- 229940124307 fluoroquinolone Drugs 0.000 description 1
- IJJVMEJXYNJXOJ-UHFFFAOYSA-N fluquinconazole Chemical compound C=1C=C(Cl)C=C(Cl)C=1N1C(=O)C2=CC(F)=CC=C2N=C1N1C=NC=N1 IJJVMEJXYNJXOJ-UHFFFAOYSA-N 0.000 description 1
- 108010006620 fodrin Proteins 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 150000002224 folic acids Chemical class 0.000 description 1
- 230000003325 follicular Effects 0.000 description 1
- 201000003444 follicular lymphoma Diseases 0.000 description 1
- 229960005102 foscarnet Drugs 0.000 description 1
- 229960004783 fotemustine Drugs 0.000 description 1
- YAKWPXVTIGTRJH-UHFFFAOYSA-N fotemustine Chemical compound CCOP(=O)(OCC)C(C)NC(=O)N(CCCl)N=O YAKWPXVTIGTRJH-UHFFFAOYSA-N 0.000 description 1
- 229940118764 francisella tularensis Drugs 0.000 description 1
- 229940044658 gallium nitrate Drugs 0.000 description 1
- 150000002270 gangliosides Chemical class 0.000 description 1
- XUBOMFCQGDBHNK-UHFFFAOYSA-N gatifloxacin Chemical compound FC1=CC(C(C(C(O)=O)=CN2C3CC3)=O)=C2C(OC)=C1N1CCNC(C)C1 XUBOMFCQGDBHNK-UHFFFAOYSA-N 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 229960005277 gemcitabine Drugs 0.000 description 1
- 229940020967 gemzar Drugs 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 230000005017 genetic modification Effects 0.000 description 1
- 235000013617 genetically modified food Nutrition 0.000 description 1
- 229940085435 giardia lamblia Drugs 0.000 description 1
- 208000005017 glioblastoma Diseases 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 150000002333 glycines Chemical class 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 125000003630 glycyl group Chemical group [H]N([H])C([H])([H])C(*)=O 0.000 description 1
- 230000001456 gonadotroph Effects 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 230000006867 granzyme B production Effects 0.000 description 1
- 231100000001 growth retardation Toxicity 0.000 description 1
- 231100000226 haematotoxicity Toxicity 0.000 description 1
- 229940047650 haemophilus influenzae Drugs 0.000 description 1
- 201000009277 hairy cell leukemia Diseases 0.000 description 1
- 201000010536 head and neck cancer Diseases 0.000 description 1
- 201000003911 head and neck carcinoma Diseases 0.000 description 1
- 208000014829 head and neck neoplasm Diseases 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 229940037467 helicobacter pylori Drugs 0.000 description 1
- 244000000013 helminth Species 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 208000002672 hepatitis B Diseases 0.000 description 1
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- UUVWYPNAQBNQJQ-UHFFFAOYSA-N hexamethylmelamine Chemical compound CN(C)C1=NC(N(C)C)=NC(N(C)C)=N1 UUVWYPNAQBNQJQ-UHFFFAOYSA-N 0.000 description 1
- 210000003630 histaminocyte Anatomy 0.000 description 1
- 102000053826 human CD70 Human genes 0.000 description 1
- 102000046492 human ICOSLG Human genes 0.000 description 1
- 102000052793 human TNFRSF14 Human genes 0.000 description 1
- 102000051198 human TNFSF14 Human genes 0.000 description 1
- 102000043656 human TNFSF15 Human genes 0.000 description 1
- 102000054539 human TNFSF8 Human genes 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 230000028996 humoral immune response Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 229960001330 hydroxycarbamide Drugs 0.000 description 1
- 229960002591 hydroxyproline Drugs 0.000 description 1
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 description 1
- 108010002685 hygromycin-B kinase Proteins 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 210000003016 hypothalamus Anatomy 0.000 description 1
- 229940015872 ibandronate Drugs 0.000 description 1
- 229960000908 idarubicin Drugs 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000005934 immune activation Effects 0.000 description 1
- 230000005746 immune checkpoint blockade Effects 0.000 description 1
- 230000037189 immune system physiology Effects 0.000 description 1
- 229940072221 immunoglobulins Drugs 0.000 description 1
- 239000000367 immunologic factor Substances 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 238000002650 immunosuppressive therapy Methods 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- DBIGHPPNXATHOF-UHFFFAOYSA-N improsulfan Chemical compound CS(=O)(=O)OCCCNCCCOS(C)(=O)=O DBIGHPPNXATHOF-UHFFFAOYSA-N 0.000 description 1
- 229950008097 improsulfan Drugs 0.000 description 1
- 238000000099 in vitro assay Methods 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 239000012678 infectious agent Substances 0.000 description 1
- 230000001524 infective effect Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 210000005007 innate immune system Anatomy 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 208000020082 intraepithelial neoplasia Diseases 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- 229960002418 ivermectin Drugs 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 229960004891 lapatinib Drugs 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 206010023841 laryngeal neoplasm Diseases 0.000 description 1
- 201000004962 larynx cancer Diseases 0.000 description 1
- 229940115932 legionella pneumophila Drugs 0.000 description 1
- 229940115286 lentinan Drugs 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 229940089519 levaquin Drugs 0.000 description 1
- 210000000088 lip Anatomy 0.000 description 1
- 229960002247 lomustine Drugs 0.000 description 1
- YROQEQPFUCPDCP-UHFFFAOYSA-N losoxantrone Chemical compound OCCNCCN1N=C2C3=CC=CC(O)=C3C(=O)C3=C2C1=CC=C3NCCNCCO YROQEQPFUCPDCP-UHFFFAOYSA-N 0.000 description 1
- 229950008745 losoxantrone Drugs 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 210000005265 lung cell Anatomy 0.000 description 1
- 210000001365 lymphatic vessel Anatomy 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- MQXVYODZCMMZEM-ZYUZMQFOSA-N mannomustine Chemical compound ClCCNC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CNCCCl MQXVYODZCMMZEM-ZYUZMQFOSA-N 0.000 description 1
- 229950008612 mannomustine Drugs 0.000 description 1
- 201000006512 mast cell neoplasm Diseases 0.000 description 1
- 208000006971 mastocytoma Diseases 0.000 description 1
- 231100000682 maximum tolerated dose Toxicity 0.000 description 1
- WKPWGQKGSOKKOO-RSFHAFMBSA-N maytansine Chemical compound CO[C@@H]([C@@]1(O)C[C@](OC(=O)N1)([C@H]([C@@H]1O[C@@]1(C)[C@@H](OC(=O)[C@H](C)N(C)C(C)=O)CC(=O)N1C)C)[H])\C=C\C=C(C)\CC2=CC(OC)=C(Cl)C1=C2 WKPWGQKGSOKKOO-RSFHAFMBSA-N 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229960004961 mechlorethamine Drugs 0.000 description 1
- HAWPXGHAZFHHAD-UHFFFAOYSA-N mechlorethamine Chemical compound ClCCN(C)CCCl HAWPXGHAZFHHAD-UHFFFAOYSA-N 0.000 description 1
- 238000002483 medication Methods 0.000 description 1
- 229960001962 mefloquine Drugs 0.000 description 1
- 210000003593 megakaryocyte Anatomy 0.000 description 1
- 229960001924 melphalan Drugs 0.000 description 1
- SGDBTWWWUNNDEQ-LBPRGKRZSA-N melphalan Chemical compound OC(=O)[C@@H](N)CC1=CC=C(N(CCCl)CCCl)C=C1 SGDBTWWWUNNDEQ-LBPRGKRZSA-N 0.000 description 1
- 210000003071 memory t lymphocyte Anatomy 0.000 description 1
- 229950009246 mepitiostane Drugs 0.000 description 1
- 229960002260 meropenem Drugs 0.000 description 1
- DMJNNHOOLUXYBV-PQTSNVLCSA-N meropenem Chemical compound C=1([C@H](C)[C@@H]2[C@H](C(N2C=1C(O)=O)=O)[C@H](O)C)S[C@@H]1CN[C@H](C(=O)N(C)C)C1 DMJNNHOOLUXYBV-PQTSNVLCSA-N 0.000 description 1
- 230000001394 metastastic effect Effects 0.000 description 1
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- VJRAUFKOOPNFIQ-TVEKBUMESA-N methyl (1r,2r,4s)-4-[(2r,4s,5s,6s)-5-[(2s,4s,5s,6s)-5-[(2s,4s,5s,6s)-4,5-dihydroxy-6-methyloxan-2-yl]oxy-4-hydroxy-6-methyloxan-2-yl]oxy-4-(dimethylamino)-6-methyloxan-2-yl]oxy-2-ethyl-2,5,7,10-tetrahydroxy-6,11-dioxo-3,4-dihydro-1h-tetracene-1-carboxylat Chemical compound O([C@H]1[C@@H](O)C[C@@H](O[C@H]1C)O[C@H]1[C@H](C[C@@H](O[C@H]1C)O[C@H]1C[C@]([C@@H](C2=CC=3C(=O)C4=C(O)C=CC(O)=C4C(=O)C=3C(O)=C21)C(=O)OC)(O)CC)N(C)C)[C@H]1C[C@H](O)[C@H](O)[C@H](C)O1 VJRAUFKOOPNFIQ-TVEKBUMESA-N 0.000 description 1
- 125000000250 methylamino group Chemical group [H]N(*)C([H])([H])[H] 0.000 description 1
- 229960003085 meticillin Drugs 0.000 description 1
- 229960000282 metronidazole Drugs 0.000 description 1
- VAOCPAMSLUNLGC-UHFFFAOYSA-N metronidazole Chemical compound CC1=NC=C([N+]([O-])=O)N1CCO VAOCPAMSLUNLGC-UHFFFAOYSA-N 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 229960004023 minocycline Drugs 0.000 description 1
- 229960005485 mitobronitol Drugs 0.000 description 1
- 230000002297 mitogenic effect Effects 0.000 description 1
- 229960003539 mitoguazone Drugs 0.000 description 1
- MXWHMTNPTTVWDM-NXOFHUPFSA-N mitoguazone Chemical compound NC(N)=N\N=C(/C)\C=N\N=C(N)N MXWHMTNPTTVWDM-NXOFHUPFSA-N 0.000 description 1
- VFKZTMPDYBFSTM-GUCUJZIJSA-N mitolactol Chemical compound BrC[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)CBr VFKZTMPDYBFSTM-GUCUJZIJSA-N 0.000 description 1
- 229950010913 mitolactol Drugs 0.000 description 1
- 229960000350 mitotane Drugs 0.000 description 1
- 101150024128 mmaA1 gene Proteins 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 201000005328 monoclonal gammopathy of uncertain significance Diseases 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 229940076266 morganella morganii Drugs 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- FABPRXSRWADJSP-MEDUHNTESA-N moxifloxacin Chemical compound COC1=C(N2C[C@H]3NCCC[C@H]3C2)C(F)=CC(C(C(C(O)=O)=C2)=O)=C1N2C1CC1 FABPRXSRWADJSP-MEDUHNTESA-N 0.000 description 1
- 108091005763 multidomain proteins Proteins 0.000 description 1
- 125000001446 muramyl group Chemical group N[C@@H](C=O)[C@@H](O[C@@H](C(=O)*)C)[C@H](O)[C@H](O)CO 0.000 description 1
- 210000000663 muscle cell Anatomy 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 239000003471 mutagenic agent Substances 0.000 description 1
- 231100000707 mutagenic chemical Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 229940055036 mycobacterium phlei Drugs 0.000 description 1
- 210000000066 myeloid cell Anatomy 0.000 description 1
- 208000025113 myeloid leukemia Diseases 0.000 description 1
- 108010065781 myosin light chain 2 Proteins 0.000 description 1
- PUPNJSIFIXXJCH-UHFFFAOYSA-N n-(4-hydroxyphenyl)-2-(1,1,3-trioxo-1,2-benzothiazol-2-yl)acetamide Chemical compound C1=CC(O)=CC=C1NC(=O)CN1S(=O)(=O)C2=CC=CC=C2C1=O PUPNJSIFIXXJCH-UHFFFAOYSA-N 0.000 description 1
- NJSMWLQOCQIOPE-OCHFTUDZSA-N n-[(e)-[10-[(e)-(4,5-dihydro-1h-imidazol-2-ylhydrazinylidene)methyl]anthracen-9-yl]methylideneamino]-4,5-dihydro-1h-imidazol-2-amine Chemical compound N1CCN=C1N\N=C\C(C1=CC=CC=C11)=C(C=CC=C2)C2=C1\C=N\NC1=NCCN1 NJSMWLQOCQIOPE-OCHFTUDZSA-N 0.000 description 1
- YOHYSYJDKVYCJI-UHFFFAOYSA-N n-[3-[[6-[3-(trifluoromethyl)anilino]pyrimidin-4-yl]amino]phenyl]cyclopropanecarboxamide Chemical compound FC(F)(F)C1=CC=CC(NC=2N=CN=C(NC=3C=C(NC(=O)C4CC4)C=CC=3)C=2)=C1 YOHYSYJDKVYCJI-UHFFFAOYSA-N 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 229940086322 navelbine Drugs 0.000 description 1
- 230000006654 negative regulation of apoptotic process Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 208000004235 neutropenia Diseases 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 229960001420 nimustine Drugs 0.000 description 1
- VFEDRRNHLBGPNN-UHFFFAOYSA-N nimustine Chemical compound CC1=NC=C(CNC(=O)N(CCCl)N=O)C(N)=N1 VFEDRRNHLBGPNN-UHFFFAOYSA-N 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 229950009266 nogalamycin Drugs 0.000 description 1
- KGTDRFCXGRULNK-JYOBTZKQSA-N nogalamycin Chemical compound CO[C@@H]1[C@@](OC)(C)[C@@H](OC)[C@H](C)O[C@H]1O[C@@H]1C2=C(O)C(C(=O)C3=C(O)C=C4[C@@]5(C)O[C@H]([C@H]([C@@H]([C@H]5O)N(C)C)O)OC4=C3C3=O)=C3C=C2[C@@H](C(=O)OC)[C@@](C)(O)C1 KGTDRFCXGRULNK-JYOBTZKQSA-N 0.000 description 1
- 230000001254 nonsecretory effect Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 201000008106 ocular cancer Diseases 0.000 description 1
- 210000004248 oligodendroglia Anatomy 0.000 description 1
- CZDBNBLGZNWKMC-MWQNXGTOSA-N olivomycin Chemical class O([C@@H]1C[C@@H](O[C@H](C)[C@@H]1O)OC=1C=C2C=C3C[C@H]([C@@H](C(=O)C3=C(O)C2=C(O)C=1)O[C@H]1O[C@@H](C)[C@H](O)[C@@H](OC2O[C@@H](C)[C@H](O)[C@@H](O)C2)C1)[C@H](OC)C(=O)[C@@H](O)[C@@H](C)O)[C@H]1C[C@H](O)[C@H](OC)[C@H](C)O1 CZDBNBLGZNWKMC-MWQNXGTOSA-N 0.000 description 1
- 201000005443 oral cavity cancer Diseases 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 229960003104 ornithine Drugs 0.000 description 1
- GSSMIHQEWAQUPM-AOLPDKKJSA-N ovalbumin peptide Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)[C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C1=CN=CN1 GSSMIHQEWAQUPM-AOLPDKKJSA-N 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 229960000625 oxytetracycline Drugs 0.000 description 1
- IWVCMVBTMGNXQD-PXOLEDIWSA-N oxytetracycline Chemical compound C1=CC=C2[C@](O)(C)[C@H]3[C@H](O)[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O IWVCMVBTMGNXQD-PXOLEDIWSA-N 0.000 description 1
- 235000019366 oxytetracycline Nutrition 0.000 description 1
- LSQZJLSUYDQPKJ-UHFFFAOYSA-N p-Hydroxyampicillin Natural products O=C1N2C(C(O)=O)C(C)(C)SC2C1NC(=O)C(N)C1=CC=C(O)C=C1 LSQZJLSUYDQPKJ-UHFFFAOYSA-N 0.000 description 1
- 101800000607 p15 Proteins 0.000 description 1
- VREZDOWOLGNDPW-UHFFFAOYSA-N pancratistatine Natural products C1=C2C3C(O)C(O)C(O)C(O)C3NC(=O)C2=C(O)C2=C1OCO2 VREZDOWOLGNDPW-UHFFFAOYSA-N 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000003154 papilloma Diseases 0.000 description 1
- 208000014837 parasitic helminthiasis infectious disease Diseases 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 229940051027 pasteurella multocida Drugs 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 229940056367 penicillin v Drugs 0.000 description 1
- 229960002340 pentostatin Drugs 0.000 description 1
- FPVKHBSQESCIEP-JQCXWYLXSA-N pentostatin Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(N=CNC[C@H]2O)=C2N=C1 FPVKHBSQESCIEP-JQCXWYLXSA-N 0.000 description 1
- QIMGFXOHTOXMQP-GFAGFCTOSA-N peplomycin Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCCN[C@@H](C)C=1C=CC=CC=1)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1NC=NC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C QIMGFXOHTOXMQP-GFAGFCTOSA-N 0.000 description 1
- 229950003180 peplomycin Drugs 0.000 description 1
- 108010044156 peptidyl-prolyl cis-trans isomerase b Proteins 0.000 description 1
- 201000002628 peritoneum cancer Diseases 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 210000003800 pharynx Anatomy 0.000 description 1
- BPLBGHOLXOTWMN-MBNYWOFBSA-N phenoxymethylpenicillin Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)COC1=CC=CC=C1 BPLBGHOLXOTWMN-MBNYWOFBSA-N 0.000 description 1
- 101150009573 phoA gene Proteins 0.000 description 1
- 238000002428 photodynamic therapy Methods 0.000 description 1
- 229960000952 pipobroman Drugs 0.000 description 1
- NJBFOOCLYDNZJN-UHFFFAOYSA-N pipobroman Chemical compound BrCCC(=O)N1CCN(C(=O)CCBr)CC1 NJBFOOCLYDNZJN-UHFFFAOYSA-N 0.000 description 1
- NUKCGLDCWQXYOQ-UHFFFAOYSA-N piposulfan Chemical compound CS(=O)(=O)OCCC(=O)N1CCN(C(=O)CCOS(C)(=O)=O)CC1 NUKCGLDCWQXYOQ-UHFFFAOYSA-N 0.000 description 1
- 229950001100 piposulfan Drugs 0.000 description 1
- 229960001221 pirarubicin Drugs 0.000 description 1
- 208000021310 pituitary gland adenoma Diseases 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 229940118768 plasmodium malariae Drugs 0.000 description 1
- 150000003057 platinum Chemical class 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229920000747 poly(lactic acid) Polymers 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 102000054765 polymorphisms of proteins Human genes 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 229960004694 prednimustine Drugs 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 229960005179 primaquine Drugs 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- CPTBDICYNRMXFX-UHFFFAOYSA-N procarbazine Chemical compound CNNCC1=CC=C(C(=O)NC(C)C)C=C1 CPTBDICYNRMXFX-UHFFFAOYSA-N 0.000 description 1
- 229960000624 procarbazine Drugs 0.000 description 1
- 230000002250 progressing effect Effects 0.000 description 1
- 208000037821 progressive disease Diseases 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000009465 prokaryotic expression Effects 0.000 description 1
- 150000003148 prolines Chemical class 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 201000001514 prostate carcinoma Diseases 0.000 description 1
- 230000009145 protein modification Effects 0.000 description 1
- 229940007042 proteus vulgaris Drugs 0.000 description 1
- WOLQREOUPKZMEX-UHFFFAOYSA-N pteroyltriglutamic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(=O)NC(CCC(=O)NC(CCC(O)=O)C(O)=O)C(O)=O)C(O)=O)C=C1 WOLQREOUPKZMEX-UHFFFAOYSA-N 0.000 description 1
- 150000003212 purines Chemical class 0.000 description 1
- 229950010131 puromycin Drugs 0.000 description 1
- 229960000996 pyrantel pamoate Drugs 0.000 description 1
- 150000003230 pyrimidines Chemical class 0.000 description 1
- JFINOWIINSTUNY-UHFFFAOYSA-N pyrrolidin-3-ylmethanesulfonamide Chemical compound NS(=O)(=O)CC1CCNC1 JFINOWIINSTUNY-UHFFFAOYSA-N 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 229960000948 quinine Drugs 0.000 description 1
- BMKDZUISNHGIBY-UHFFFAOYSA-N razoxane Chemical compound C1C(=O)NC(=O)CN1C(C)CN1CC(=O)NC(=O)C1 BMKDZUISNHGIBY-UHFFFAOYSA-N 0.000 description 1
- 229960000460 razoxane Drugs 0.000 description 1
- 230000007420 reactivation Effects 0.000 description 1
- 101150079601 recA gene Proteins 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000007115 recruitment Effects 0.000 description 1
- 108010054624 red fluorescent protein Proteins 0.000 description 1
- 239000003488 releasing hormone Substances 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 229930002330 retinoic acid Natural products 0.000 description 1
- 201000009410 rhabdomyosarcoma Diseases 0.000 description 1
- OWPCHSCAPHNHAV-LMONGJCWSA-N rhizoxin Chemical compound C/C([C@H](OC)[C@@H](C)[C@@H]1C[C@H](O)[C@]2(C)O[C@@H]2/C=C/[C@@H](C)[C@]2([H])OC(=O)C[C@@](C2)(C[C@@H]2O[C@H]2C(=O)O1)[H])=C\C=C\C(\C)=C\C1=COC(C)=N1 OWPCHSCAPHNHAV-LMONGJCWSA-N 0.000 description 1
- 229950004892 rodorubicin Drugs 0.000 description 1
- MBABCNBNDNGODA-WPZDJQSSSA-N rolliniastatin 1 Natural products O1[C@@H]([C@@H](O)CCCCCCCCCC)CC[C@H]1[C@H]1O[C@@H]([C@H](O)CCCCCCCCCC[C@@H](O)CC=2C(O[C@@H](C)C=2)=O)CC1 MBABCNBNDNGODA-WPZDJQSSSA-N 0.000 description 1
- IMUQLZLGWJSVMV-UOBFQKKOSA-N roridin A Natural products CC(O)C1OCCC(C)C(O)C(=O)OCC2CC(=CC3OC4CC(OC(=O)C=C/C=C/1)C(C)(C23)C45CO5)C IMUQLZLGWJSVMV-UOBFQKKOSA-N 0.000 description 1
- VHXNKPBCCMUMSW-FQEVSTJZSA-N rubitecan Chemical compound C1=CC([N+]([O-])=O)=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VHXNKPBCCMUMSW-FQEVSTJZSA-N 0.000 description 1
- 201000003804 salivary gland carcinoma Diseases 0.000 description 1
- 235000019515 salmon Nutrition 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 229930182947 sarcodictyin Natural products 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 229940055619 selenocysteine Drugs 0.000 description 1
- 235000016491 selenocysteine Nutrition 0.000 description 1
- ZKZBPNGNEQAJSX-UHFFFAOYSA-N selenocysteine Natural products [SeH]CC(N)C(O)=O ZKZBPNGNEQAJSX-UHFFFAOYSA-N 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000000717 sertoli cell Anatomy 0.000 description 1
- 229940007046 shigella dysenteriae Drugs 0.000 description 1
- 229940115939 shigella sonnei Drugs 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 239000013605 shuttle vector Substances 0.000 description 1
- 230000007781 signaling event Effects 0.000 description 1
- 238000009097 single-agent therapy Methods 0.000 description 1
- 210000002027 skeletal muscle Anatomy 0.000 description 1
- 201000000849 skin cancer Diseases 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 1
- 229940079832 sodium starch glycolate Drugs 0.000 description 1
- 229920003109 sodium starch glycolate Polymers 0.000 description 1
- 239000008109 sodium starch glycolate Substances 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 210000000278 spinal cord Anatomy 0.000 description 1
- 206010062261 spinal cord neoplasm Diseases 0.000 description 1
- 229950006315 spirogermanium Drugs 0.000 description 1
- ICXJVZHDZFXYQC-UHFFFAOYSA-N spongistatin 1 Natural products OC1C(O2)(O)CC(O)C(C)C2CCCC=CC(O2)CC(O)CC2(O2)CC(OC)CC2CC(=O)C(C)C(OC(C)=O)C(C)C(=C)CC(O2)CC(C)(O)CC2(O2)CC(OC(C)=O)CC2CC(=O)OC2C(O)C(CC(=C)CC(O)C=CC(Cl)=C)OC1C2C ICXJVZHDZFXYQC-UHFFFAOYSA-N 0.000 description 1
- 229940031439 squalene Drugs 0.000 description 1
- TUHBEKDERLKLEC-UHFFFAOYSA-N squalene Natural products CC(=CCCC(=CCCC(=CCCC=C(/C)CCC=C(/C)CC=C(C)C)C)C)C TUHBEKDERLKLEC-UHFFFAOYSA-N 0.000 description 1
- 208000017572 squamous cell neoplasm Diseases 0.000 description 1
- 230000010473 stable expression Effects 0.000 description 1
- 229940037649 staphylococcus haemolyticus Drugs 0.000 description 1
- 229940071117 starch glycolate Drugs 0.000 description 1
- 239000000021 stimulant Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 229940031000 streptococcus pneumoniae Drugs 0.000 description 1
- 229960001052 streptozocin Drugs 0.000 description 1
- ZSJLQEPLLKMAKR-GKHCUFPYSA-N streptozocin Chemical compound O=NN(C)C(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O ZSJLQEPLLKMAKR-GKHCUFPYSA-N 0.000 description 1
- 201000009032 substance abuse Diseases 0.000 description 1
- 231100000736 substance abuse Toxicity 0.000 description 1
- 208000011117 substance-related disease Diseases 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 230000000153 supplemental effect Effects 0.000 description 1
- 230000003319 supportive effect Effects 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 229940126625 tavolimab Drugs 0.000 description 1
- 229940063683 taxotere Drugs 0.000 description 1
- 229940066453 tecentriq Drugs 0.000 description 1
- 108091035539 telomere Proteins 0.000 description 1
- NRUKOCRGYNPUPR-QBPJDGROSA-N teniposide Chemical compound COC1=C(O)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@@H](O[C@H]3[C@@H]([C@@H](O)[C@@H]4O[C@@H](OC[C@H]4O3)C=3SC=CC=3)O)[C@@H]3[C@@H]2C(OC3)=O)=C1 NRUKOCRGYNPUPR-QBPJDGROSA-N 0.000 description 1
- 229960001278 teniposide Drugs 0.000 description 1
- 229940061354 tequin Drugs 0.000 description 1
- IWVCMVBTMGNXQD-UHFFFAOYSA-N terramycin dehydrate Natural products C1=CC=C2C(O)(C)C3C(O)C4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O IWVCMVBTMGNXQD-UHFFFAOYSA-N 0.000 description 1
- 230000002381 testicular Effects 0.000 description 1
- 201000003120 testicular cancer Diseases 0.000 description 1
- BPEWUONYVDABNZ-DZBHQSCQSA-N testolactone Chemical compound O=C1C=C[C@]2(C)[C@H]3CC[C@](C)(OC(=O)CC4)[C@@H]4[C@@H]3CCC2=C1 BPEWUONYVDABNZ-DZBHQSCQSA-N 0.000 description 1
- 229960005353 testolactone Drugs 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 229940072172 tetracycline antibiotic Drugs 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- 230000004797 therapeutic response Effects 0.000 description 1
- 201000002510 thyroid cancer Diseases 0.000 description 1
- YFTWHEBLORWGNI-UHFFFAOYSA-N tiamiprine Chemical compound CN1C=NC([N+]([O-])=O)=C1SC1=NC(N)=NC2=C1NC=N2 YFTWHEBLORWGNI-UHFFFAOYSA-N 0.000 description 1
- 229950011457 tiamiprine Drugs 0.000 description 1
- 229960005053 tinidazole Drugs 0.000 description 1
- 210000002105 tongue Anatomy 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 229940044693 topoisomerase inhibitor Drugs 0.000 description 1
- 229960000303 topotecan Drugs 0.000 description 1
- UCFGDBYHRUNTLO-QHCPKHFHSA-N topotecan Chemical compound C1=C(O)C(CN(C)C)=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 UCFGDBYHRUNTLO-QHCPKHFHSA-N 0.000 description 1
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 206010044412 transitional cell carcinoma Diseases 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 108091007466 transmembrane glycoproteins Proteins 0.000 description 1
- 230000000472 traumatic effect Effects 0.000 description 1
- 229950001353 tretamine Drugs 0.000 description 1
- IUCJMVBFZDHPDX-UHFFFAOYSA-N tretamine Chemical compound C1CN1C1=NC(N2CC2)=NC(N2CC2)=N1 IUCJMVBFZDHPDX-UHFFFAOYSA-N 0.000 description 1
- 229960001727 tretinoin Drugs 0.000 description 1
- 229960004560 triaziquone Drugs 0.000 description 1
- PXSOHRWMIRDKMP-UHFFFAOYSA-N triaziquone Chemical compound O=C1C(N2CC2)=C(N2CC2)C(=O)C=C1N1CC1 PXSOHRWMIRDKMP-UHFFFAOYSA-N 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 229930013292 trichothecene Natural products 0.000 description 1
- 150000003327 trichothecene derivatives Chemical class 0.000 description 1
- 229960001670 trilostane Drugs 0.000 description 1
- KVJXBPDAXMEYOA-CXANFOAXSA-N trilostane Chemical compound OC1=C(C#N)C[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CC[C@@]32O[C@@H]31 KVJXBPDAXMEYOA-CXANFOAXSA-N 0.000 description 1
- 229960001099 trimetrexate Drugs 0.000 description 1
- NOYPYLRCIDNJJB-UHFFFAOYSA-N trimetrexate Chemical compound COC1=C(OC)C(OC)=CC(NCC=2C(=C3C(N)=NC(N)=NC3=CC=2)C)=C1 NOYPYLRCIDNJJB-UHFFFAOYSA-N 0.000 description 1
- 229960000875 trofosfamide Drugs 0.000 description 1
- UMKFEPPTGMDVMI-UHFFFAOYSA-N trofosfamide Chemical compound ClCCN(CCCl)P1(=O)OCCCN1CCCl UMKFEPPTGMDVMI-UHFFFAOYSA-N 0.000 description 1
- HDZZVAMISRMYHH-LITAXDCLSA-N tubercidin Chemical compound C1=CC=2C(N)=NC=NC=2N1[C@@H]1O[C@@H](CO)[C@H](O)[C@H]1O HDZZVAMISRMYHH-LITAXDCLSA-N 0.000 description 1
- 230000005740 tumor formation Effects 0.000 description 1
- 230000005909 tumor killing Effects 0.000 description 1
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 1
- 229940094060 tykerb Drugs 0.000 description 1
- 229950009811 ubenimex Drugs 0.000 description 1
- 231100000402 unacceptable toxicity Toxicity 0.000 description 1
- 230000004222 uncontrolled growth Effects 0.000 description 1
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 1
- 241001529453 unidentified herpesvirus Species 0.000 description 1
- 241000712461 unidentified influenza virus Species 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 229960001055 uracil mustard Drugs 0.000 description 1
- 210000003932 urinary bladder Anatomy 0.000 description 1
- 230000002485 urinary effect Effects 0.000 description 1
- 206010046766 uterine cancer Diseases 0.000 description 1
- 208000024719 uterine cervix neoplasm Diseases 0.000 description 1
- 208000007089 vaccinia Diseases 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 229960003165 vancomycin Drugs 0.000 description 1
- MYPYJXKWCTUITO-LYRMYLQWSA-N vancomycin Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-N 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 229940118696 vibrio cholerae Drugs 0.000 description 1
- 229960003048 vinblastine Drugs 0.000 description 1
- JXLYSJRDGCGARV-XQKSVPLYSA-N vincaleukoblastine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](OC(C)=O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(=O)OC)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 JXLYSJRDGCGARV-XQKSVPLYSA-N 0.000 description 1
- 229960004528 vincristine Drugs 0.000 description 1
- OGWKCGZFUXNPDA-UHFFFAOYSA-N vincristine Natural products C1C(CC)(O)CC(CC2(C(=O)OC)C=3C(=CC4=C(C56C(C(C(OC(C)=O)C7(CC)C=CCN(C67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)CN1CCC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-UHFFFAOYSA-N 0.000 description 1
- OGWKCGZFUXNPDA-XQKSVPLYSA-N vincristine Chemical compound C([N@]1C[C@@H](C[C@]2(C(=O)OC)C=3C(=CC4=C([C@]56[C@H]([C@@]([C@H](OC(C)=O)[C@]7(CC)C=CCN([C@H]67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)C[C@@](C1)(O)CC)CC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-XQKSVPLYSA-N 0.000 description 1
- 229960004355 vindesine Drugs 0.000 description 1
- UGGWPQSBPIFKDZ-KOTLKJBCSA-N vindesine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(N)=O)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1N=C1[C]2C=CC=C1 UGGWPQSBPIFKDZ-KOTLKJBCSA-N 0.000 description 1
- GBABOYUKABKIAF-IELIFDKJSA-N vinorelbine Chemical compound C1N(CC=2C3=CC=CC=C3NC=22)CC(CC)=C[C@H]1C[C@]2(C(=O)OC)C1=CC([C@]23[C@H]([C@@]([C@H](OC(C)=O)[C@]4(CC)C=CCN([C@H]34)CC2)(O)C(=O)OC)N2C)=C2C=C1OC GBABOYUKABKIAF-IELIFDKJSA-N 0.000 description 1
- 229960002066 vinorelbine Drugs 0.000 description 1
- CILBMBUYJCWATM-PYGJLNRPSA-N vinorelbine ditartrate Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O.OC(=O)[C@H](O)[C@@H](O)C(O)=O.C1N(CC=2C3=CC=CC=C3NC=22)CC(CC)=C[C@H]1C[C@]2(C(=O)OC)C1=CC([C@]23[C@H]([C@@]([C@H](OC(C)=O)[C@]4(CC)C=CCN([C@H]34)CC2)(O)C(=O)OC)N2C)=C2C=C1OC CILBMBUYJCWATM-PYGJLNRPSA-N 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
- 201000005102 vulva cancer Diseases 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 238000011816 wild-type C57Bl6 mouse Methods 0.000 description 1
- 229940053867 xeloda Drugs 0.000 description 1
- 229940051021 yellow-fever virus Drugs 0.000 description 1
- 229940098232 yersinia enterocolitica Drugs 0.000 description 1
- 229940055760 yervoy Drugs 0.000 description 1
- 229950009268 zinostatin Drugs 0.000 description 1
- 229960000641 zorubicin Drugs 0.000 description 1
- FBTUMDXHSRTGRV-ALTNURHMSA-N zorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(\C)=N\NC(=O)C=1C=CC=CC=1)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 FBTUMDXHSRTGRV-ALTNURHMSA-N 0.000 description 1
Classifications
-
- 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
- C07K14/70575—NGF/TNF-superfamily, e.g. CD70, CD95L, CD153, CD154
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
- A61K2239/31—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterized by the route of administration
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
- A61K2239/38—Indexing codes associated with cellular immunotherapy of group A61K39/46 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
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
- A61K39/001176—Heat shock proteins
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/461—Cellular immunotherapy characterised by the cell type used
- A61K39/4611—T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/4643—Vertebrate antigens
- A61K39/4644—Cancer antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
- 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
-
- 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
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/515—Animal cells
- A61K2039/5152—Tumor cells
-
- 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
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55516—Proteins; Peptides
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Definitions
- the disclosure is directed to methods of treatment with cells having a vaccine ⁇ e.g., gp96-lg) and cells having T-cell co-stimulatory molecules.
- Cancer is characterized by a progressive acquisition of genetic mutations that lead to intrinsic dysregulation of cell growth and death. Once a cell has acquired enough mutations, typically thought to be at least six, it no longer is responsive to intrinsic or extrinsic signals that would restrain its growth or trigger apoptosis. As tumors arise from host cells, the body's immune system is initially tolerant to those cells. A cell that acquires an immunogenic mutation, can be sought out and destroyed by the host immune system in a process known as immunosurveillance. Immune checkpoint therapy, which targets regulatory pathways in T cells to enhance anti-tumor immune responses, has led to important clinical advances and provides a new defense against cancer. Moreover, vaccines may contribute to this defense by also enhancing anti-tumor immune responses. Accordingly, it is possible that combination therapies including combinations or sub-combinations of one or more checkpoint inhibitors, one or more vaccines, and one or more T cell costimulatory molecules may expand the base of cancer patients that can benefit from immunotherapy.
- Immunotherapies aimed at including combinations of one or more vaccines, one or more T cell costimulatory molecules, and one or more checkpoint inhibitors may expand the base of cancer patients that can benefit from such therapies.
- Vaccines may contribute to this response by increasing both the frequency of tumor-antigen specific CD8+ T cells and also the number of tumor antigens recognized by those CD8+ T cells.
- T cell costimulatory molecules may enhance the response by further increasing the frequency and/or enhancing the activation of tumor antigen-specific T cells, and also by increasing the expression of tumor-killing effector molecules by CD8+T cells.
- a combination of a vaccination, e.g., gp96-lg vaccination, and T cell costimulation with one or more agonists of 0X40, ICOS, 4-1 BB, TNFRSF25, CD40, CD27, and/or GITR, among others, provides a synergistic anti-tumor benefit.
- Pre- clinical models have evaluated independent compositions of gp96-lg vaccines combined with agonistic antibodies targeting 0X40, ICOS, 4-1 BB, and TNFRSF25, and demonstrated variable effects on mechanistic and anti-tumor complementarity.
- the methods described herein provide a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein, ⁇ e.g., gp96-lg) expression vector, wherein the patient is undergoing a treatment with a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein, including, without limitation, fusion proteins such as ICOSL-lg, 4-1 BBL-lg, TL1 A-lg, OX40L-lg, CD40L-lg, CD70-lg, or GITRL-lg to provide T cell costimulation.
- fusion proteins such as ICOSL-lg, 4-1 BBL-lg, TL1 A-lg, OX40L-lg, CD40L-lg, CD70-lg, or GITRL-lg to provide T cell costimulation.
- the methods described herein secrete fusion proteins that work synergistically.
- the effect of a locally secreted T-cell costimulatory fusion protein i.e., OX40L-lg
- OX40L-lg performs differently than a systemic administration in combination with the secretable vaccine protein (e.g., gp96-lg).
- the effects of a cell secreting a vaccine protein alone [e.g., gp96-lg) and in combination with escalating doses of a cell secreting a T-cell costimulatory fusion protein ⁇ i.e., OX40L-lg), performs differently when compared to the vaccine protein (e.g., gp96-lg) and escalating doses of a systemic 0X40 agonist antibody.
- the amount of a secretable vaccine protein (e.g., gp96-lg) secretion is higher than the expression of a T cell costimulatory fusion protein, (e.g., OX40L-lg).
- the ratio of a vaccine protein (e.g., gp96-lg) secretion to the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) is about 1 :10, 1 :25, 1 :50, 1 :100, 1 :200, 1 :300, 1 :400, 1 :500, 1 :600, 1 :700, 1 :800, 1 :900 or 1 :1000, (inclusive of all endpoints).
- the amount of a vaccine protein (e.g., gp96-lg) secretion is lower than the expression of a T cell costimulatory fusion protein ⁇ e.g., OX40L-lg).
- the ratio of a vaccine protein (e.g., gp96-lg) secretion to the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) is about 10:1 , 25:1 , 50:1 , 100:1 , 200:1 , 300:1 , 400:1 , 500:1 , 600:1 , 700:1 , 800:1 , 900:1 , or 1000:1 , (inclusive of all endpoints).
- the amount of the expression of a T cell costimulatory fusion protein is higher than the secretion of a vaccine protein (e.g., gp96-lg).
- the ratio of the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) to the secretion of a vaccine protein (e.g., gp96-lg) is about 1 :10, 1 :25, 1 :50, 1 :100, 1 :200, 1 :300, 1 :400, 1 :500, 1 :600, 1 :700, 1 :800, 1 :900, or 1 :1000, (inclusive of all endpoints).
- the amount of the expression of a T cell costimulatory fusion protein is lower than the secretion of a vaccine protein (e.g., gp96-lg).
- the ratio of the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) to the secretion of a vaccine protein (e.g., gp96-lg) is about 10:1 , 25:1 , 50:1 , 100:1 , 200:1 , 300:1 , 400:1 , 500:1 , 600:1 , 700:1 , 800:1 , 900:1 , or 1000:1 , (inclusive of all endpoints).
- the amount of a secretable vaccine protein (e.g., gp96-lg) secretion is about the same as the expression of a T cell costimulatory fusion protein, (e.g., OX40L-lg).
- the ratio of a vaccine protein (e.g., gp96-lg) secretion to the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) is about 1 :1.
- the ratio of a vaccine protein (e.g., gp96-lg) secretion to the expression of a T cell costimulatory fusion protein, such as OX40L- Ig is about 1 :1.3.
- the amount of a secretable vaccine protein (e.g., gp96-lg) expression is about the same as the expression of a T cell costimulatory fusion protein, (e.g., OX40L-lg).
- the ratio of a vaccine protein (e.g., gp96-lg) expression to the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) is about 1 :1.
- the ratio of a vaccine protein (e.g., gp96-lg) expression to the expression of a T cell costimulatory fusion protein, such as OX40L-lg is about 1 :1 .3.
- the amount of the expression of a T cell costimulatory fusion protein is about the same as the secretion of a vaccine protein (e.g., gp96-lg).
- the ratio of the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) to the secretion of a vaccine protein (e.g., gp96-lg) is about 1 :1.
- the number of cells secreting a gp96-lg is higher than the number of cells secreting a OX40L-lg.
- the ratio of the number of cells secreting a gp96-lg to the number of cells secreting a OX40L-lg is about 1: 0.01, about 1: 0.1, about 1: 1, about 1:10, about 1:25, about 1:50, about 1:100, about 1:200, about 1:300, about 1:400, about 1:500, about 1:600, about 1:700, about 1 :800, about 1 :900 or about 1 :1000 (inclusive of all endpoints).
- the number of cells secreting a gp96-lg is lower than the number of cells secreting a OX40L-lg.
- the ratio of the number of cells secreting a gp96-lg to the number of cells secreting a OX40L-lg is about 0.01: 1, about 0.1: 1, about 1:1, about 1:1.3, about 10:1, about 25:1, about 50:1, about 100:1, about 200:1, about 300:1, about 400:1, about 500:1, about 600:1, about 700:1, about 800:1, about 900:1, or about 1000:1 (inclusive of all endpoints).
- the expression of a gp96-lg is higher than the expression of a OX40L-lg.
- the ratio of the expression of the gp96-lg to the expression of the OX40L-lg is about 1: 0.01, about 1: 0.1, about 1: 1, about 1:10, about 1:25, about 1:50, about 1:100, about 1:200, about 1:300, about 1:400, about 1:500, about 1:600, about 1:700, about 1:800, about 1:900 or about 1:1000 (inclusive of all endpoints).
- the expression of a gp96-lg is lower than the expression of a OX40L-lg.
- the ratio of the expression of a gp96-lg to the expression of a OX40L-lg is about 0.01: 1, aboutO.1: 1, about 1:1, about 1:3, about 10:1, about 25:1, about 50:1, about 100:1, about 200:1, about 300:1, about 400:1, about 500:1, about 600:1, about 700:1, about 800:1, about 900:1, or about 1000:1 (inclusive of all endpoints).
- an inducible promoter can be used for inducing the expression of the vaccine protein ⁇ e.g., gp96-lg).
- the gp96-lg is under a strong inducible promoter.
- the gp96-lg is under an intermediate inducible promoter.
- the gp96-lg is under a weak inducible promoter.
- an inducible promoter can be used for inducing the expression T cell costimulatory fusion ⁇ e.g., OX40L-lg).
- the OX40L-lg is under a strong inducible promoter.
- the OX40L-lg is under an intermediate inducible promoter.
- the OX40L-lg is under a weak inducible promoter.
- the vaccine protein e.g., gp96-lg
- T cell costimulatory fusion protein e.g., OX40L-lg
- host cells e.g., mammalian cells
- expression and/or secretion of the gp96-lg and/or OX40L-lg can be readily detected and quantified by techniques known in the art, such as, in vitro cell culturing methods or protein detection assays.
- the protein detection assays include enzyme-linked immunosorbent assay (ELISA), immunoprecipitation, and fluorescence based methods.
- the amount of secreted gp96-lg by a cell is higher than the amount of secreted OX40L-lg by a cell.
- the ratio of the secreted gp96-lg by a cell to the secreted OX40L-lg by a cell is about 1 : 0.01 , about 1 : 0.1 , about 1 : 1 , about 1 :10, about 1 :25, about 1 :50, about 1 :100, about 1 :200, about 1 :300, about 1 :400, about 1 :500, about 1 :600, about 1 :700, about 1 :800, about 1 :900 or about 1 :1000 (inclusive of all endpoints).
- the amount of secreted gp96-lg by a cell is lower than the amount of secreted OX40L-lg by a cell.
- the ratio of secreted gp96-lg by a cell to secreted OX40L-lg by a cell is about 0.01 : 1 , about 0.1 : 1 , about 1 :1 , about 1 :1.3, about 10:1 , about 25:1 , about 50:1 , about 100:1 , about 200:1 , about 300:1 , about 400:1 , about 500:1 , about 600:1 , about 700:1 , about 800:1 , about 900:1 , or about 1000:1 (inclusive of all endpoints).
- the disclosure provides a method for treating a patient comprising administering to the patient an effective amount of a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein, wherein the patient is undergoing a treatment with a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject.
- the disclosure provides a method for treating a patient comprising administering to the patient an effective amount of a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein, wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject, and wherein the patient is undergoing a treatment with a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein.
- the disclosure provides a method for treating a patient comprising administering to the patient an effective amount of (a) a first cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and (b) a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject.
- the secretable vaccine protein is a secretable gp96-lg fusion protein which optionally lacks the gp96 KDEL (SEQ ID NO:3) sequence.
- the Ig tag in the gp96-lg fusion protein comprises the Fc region of human lgG1 , lgG2, lgG3, lgG4, IgM, IgA, or IgE.
- the T cell costimulatory fusion protein is OX40L-lg, or a portion thereof that binds to 0X40. In some embodiments, the T cell costimulatory fusion protein is ICOSL-lg, or a portion thereof that binds to ICOS. In some embodiments, the T cell costimulatory fusion protein is 4-1 BBL-lg, or a portion thereof that binds to 4-1 BBR. In some embodiments, the T cell costimulatory fusion protein is TL1 A-lg, or a portion thereof that binds to TNFRSF25.
- the T cell costimulatory fusion protein is GITRL-lg, or a portion thereof that binds to GITR. In some embodiments, the T cell costimulatory fusion protein is CD40L-lg, or a portion thereof that binds to CD40. In some embodiments, the T cell costimulatory fusion protein is CD70-lg, or a portion thereof that binds to CD27. In some embodiments, the Ig tag in the T cell costimulatory fusion protein comprises the Fc region of human lgG1 , lgG2, lgG3, lgG4, IgM, IgA, or IgE.
- the expression vector is incorporated into a virus or virus-like particle. In some embodiments, the expression vector is incorporated into a human tumor cell. In some embodiments, the patient is a human cancer patient. In some embodiments, administration to the human patient increases the activation or proliferation of tumor antigen specific T cells in the patient.
- the activation or proliferation of tumor antigen specific T cells in the patient is increased by at least 25 percent as compared to the level of activation or proliferation of tumor antigen specific T cells in the patient prior to the administration.
- administration is in combination with an agent that inhibits immunosuppressive molecules produced by tumor cells.
- the agent is an antibody against PD-1.
- the antibody against PD-1 is selected from Nivolumab, Pembrolizumab, Pidilizumab, Cemiplimab, AGEN2034, AMP-224, AMP-514, PDR001.
- the patient is a human with an acute or chronic infection.
- the acute or chronic infection is an infection by hepatitis C virus, hepatitis B virus, human immunodeficiency virus, or malaria.
- administration to the human patient stimulates the activation or proliferation of pathogenic antigen specific T cells.
- the T cell costimulatory molecule enhances the activation of antigen- specific T cells in the subject to a greater level than gp96-lg vaccination alone.
- FIG. 1 is a plasmid vector map for pcDNA3.4 OX40L-lg.
- FIG. 2 is a graph showing the activation of human 0X40 receptor in Jurkat cells by mouse and human OX40L.
- FIG. 3 is an image showing mouse HS-110 (B16F10-OVA-gp96) and mouse HS-130 (B16F10- OVA- OX40L) immunization over a dose-range to correlate CD8+T-cell expansion to tumor growth delay.
- FIG. 4 shows flow cytometry diagrams, dot plots and gating strategy for Day 7 after a primary vaccination (peripheral blood). Recipient mice were injected with a mouse mHS-110 at a constant dose of 1 million cells (290 ng of gp96-lg) with different ratios of mHS-130 (0.1 , 0.3, 1 , 3, 10).
- FIG. 4 shows a flow cytometry gating strategy using FlowJo Version 10 (2018), by gating in the blood on singlets and CD3+ T cells, then CD8+ OT-I GFP+ T cells. Sample analysis was taken on day 7 and numbers in representative dot plots indicate percentages of CD8+ OT-I GFP+ positive cells within the gated population. Plots show an individual, representative mouse that illustrates peak expansion for the day selected.
- FIG. 5 shows flow cytometry diagrams, dot plots and gating strategy for Day 21 after a boost vaccination (peripheral blood).
- Recipient mice were injected with a mouse mHS-110 at a constant dose of 1 million cells (290 ng of gp96-lg) with different ratios of mHS-130 (0.1 , 0.3, 1 , 3, 10).
- a ratio of 1 to 1 is 290 ng of gp96 (1 million mHS-110 cells) to 290 ng of OX40L.
- FIG. 5 shows a flow cytometry gating strategy using FlowJo Version 10 (2018), by gating in the blood on singlets and CD3+ T cells, then CD8+ OT-I GFP+ T cells.
- FIG. 6A and FIG. 6B are graphs showing the percent of OT-I CD8+ T-cells after primary and secondary vaccination with a set dose mHS-110 with different ratios of mHS-130 before and after tumor challenge (peripheral blood).
- Recipient mice were injected with a mHS-110 at a constant dose of 1 million cells (290 ng of gp96-lg) with different ratios of mHS130.
- OT-I GFP+ CD8+T cells were analyzed in the blood on days 0-53 days post-vaccination.
- mice were boosted on day 14 with the same ratios of mHS110 and mHS130 as in the primary phase, and OT-I GFP+ CD8+ T cells were analyzed in the blood on days 17, 19, 21 , 24, 28, 33, 38, 41 days post-challenge.
- FIG. 6A Line graph without overlay
- FIG. 6B Line graph with mouse outliers removed out to day 41 only.
- FIG. 7A, FIG. 7B, FIG. 7C, FIG. 7D, and FIG. 7E are graphs showing the day-54, End-of-Study, endogenous response to vaccination.
- FIG. 7A is a graph showing an End-of study, day-54, endogenous spleen response to vaccination, percent, gated FSC-H by FSC-A as to live/dead gate for doublets, then gated on CD45 by SSC then CD3+ CD8+ double positive cells.
- FIG. 7A shows mean ⁇ SEM, * p ⁇ 0.05 compared to mHS-110 via the non-parameteric statistical test, Mann-Whitney.
- FIG. 7B is a flow cytometry diagram and FIG.
- FIG. 7C shows a graph of End-of study, day-54, endogenous spleen response to vaccination and ex vivo stimulation with pg100 peptide for intracellular cytokine staining. Percent shown in graph. Events gated on FSC-H by FSC-A as to live/dead gate for doublets, then gated on CD45 by SSC then IFN-g CD8+ double positive cells.
- FIG. 7C is a graph showing mean ⁇ SEM, *p ⁇ 0.05 compared to mHS-110 via the non-parameteric statistical test, Mann-Whitney;‘ns’ denotes p>0.05, not significant.
- FIG. 7D are graphs showing endogenous spleen immune response measured by IFN-gamma ELISPOT.
- FIG. 7D shows graphs having mean ⁇ SEM IFN-g spots per million splenocytes, **p ⁇ 0.01 , *p ⁇ 0.05 compared to mHS-1 10 via the non-parameteric statistical test, Mann-Whitney;‘ns’ denotes p>0.05, not significant.
- Far right shows representative ELISPOT well with machine counts. Background (media alone) wells not subtracted from graphed data sets. Positive control wells worked (data not shown).
- FIG. 7E is a graph showing End-of study, day-54, endogenous response to vaccination, percent, gated FSC-H by FSC-A as to live/dead gate for doublets, then gated on CD45 by SSC then CD3+ CD4+ double positive cells.
- FIG. 7E shows mean ⁇ SEM, **p ⁇ 0.01 compared to mHS-1 10 via the non-parameteric statistical test, Mann-Whitney.
- FIG. 8A is a flow cytometry diagram and FIG. 8B is a graph showing CD8+ Tumor Infiltrating Lymphocytes (TILs).
- TILs Tumor Infiltrating Lymphocytes
- FIG. 8A shows graphs of the End-of study, day-54, endogenous TIL response to vaccination, percent, gated FSC-H by FSC-A to as live/dead gate for doublets, then gated on CD45 by SSC then CD3+ CD8+ double positive cells.
- the MACS Miltenyl Biotec tumor dissociation kit was used for this procedure (cat# 130-096-730).
- FIG. 8B is a graph showing mean ⁇ SEM, *p ⁇ 0.05 compared to mHS-1 10 via the non-parameteric statistical test, Mann-Whitney.
- FIG. 9 is a graph showing End of study, final tumor mass, in grams, of individual mice. Tumor mass (wet weight) were weighed using a milligram sensitive scale for each animal. FIG. 9 shows mean ⁇ SEM. Statistics performed were non-parametric Mann-Whitney, ‘ns’ designates a non-significant (p>0.05) value, *p ⁇ 0.05; **p ⁇ 0.01.
- FIG.10A and FIG. 10B shows tumor volumes over time, tumor mean size, and individual plots for individual animals.
- FIG. 10A is a graph showing mean ⁇ SEM for all tumor volumes over time.
- FIG. 10B is a graph showing means for individual animals for each measured timepoint.
- Tumor implantation, melanoma B16F10 cells were harvested and resuspended at a concentration of 5x10 5 cells/1 OOpl in a volume of 80mI HBSS and 20pl Matrigel.
- C57BL/6 mice were subcutaneously injected with 100 mI of B16F10 cells (5 x 10 5 cells/mouse) on the inner abdomen .
- the tumor size was measured and documented every 3 days with a caliper, starting on day 7, and calculated using the formula (A * B) (A as the largest and B as the smallest diameter of tumor). Tumor growth was documented as standard error mean. To record the survival of the tumor-bearing mice, either natural death or a tumor volume greater than 450 mm 2 leading to death was counted as death. Each experimental group included five animals. Statistics performed for FIG. 10A was a 2-way ANOVA (shown below FIG. 10A), p ⁇ 0.05 is considered significantly different vs. mHS-1 10 group alone.
- FIG. HA and FIG. 11 B are graphs showing the percent CD8+ OT-1 + T-cells on day 54 (spleen), and the flow plot gating strategy.
- FIG. 11 A is a graph showing the End-of study, day-54, endogenous response to vaccination, percent, gated FSC-H by FSC-A as to live/dead gate for doublets, then gated on CD45 by SSC then GFP-OT-1 CD8+ double positive cells.
- FIG. 11 B is a graph showing mean ⁇ SEM, *p ⁇ 0.05 compared to mHS-110 via the non-parametric statistical test, Mann-Whitney.
- FIG. 12A and FIG. 12B are graphs showing the percent CD8+ PD-1 + T-cells on day 54 (spleen), and the flow plot gating strategy.
- FIG. 12A is a graph showing End-of study, day-54, endogenous response to vaccination, percent, gated FSC-H by FSC-A as to live/dead gate for doublets, then gated on CD3 by SSC, then PD-1 + CD8+ double positive cells.
- FIG. 12B is a graph showing mean ⁇ SEM, *p ⁇ 0.05 compared to mHS-110 via the non-parameteric statistical test, Mann-Whitney.
- FIG. 13 is a non-limiting schematic of the design of the study of gp96-lg (mHS-110, B16F10- OVA-gp96) to OX40L-lg (mHS-130, B16F10-OVA- OX40L) dose ratios, to correlate CD8+ T-cell expansion to tumor growth delay.
- FIG. 14 are graphs illustrating anti-tumor CD8+ OT-I T cell expansion in the peripheral blood with prime and boost Immunization of different ratios and dose combinations of mHS-1 10 and mHS-130, in the study of FIG. 13.
- Recipient mice were injected with mHS-1 10 and mHS-130 at different ratios and doses of gp96-lg to OX40L-lg.
- OT-I GFP+ CD8+ T cells were analyzed in the blood on days 0-54 days post-vaccination.
- mice were boosted on day 14 with the same ratios of mHS-110 and mHS-130 as in the primary phase, and OT-I GFP+ CD8+ T cells were analyzed in the blood days post-challenge. Data represent mean percent ⁇ SEM.
- FIGS. 15A to 15D illustrate flow cytometry gating strategy and cellular expansion of CD8+ OT-I T-cells, over time, with mHS-110/130 immunization in the study of FIG. 13.
- FIG. 15A are graphs illustrating flow cytometry gating strategy of CD8+ OT-I T-cells, over time, for the tested ratios and mPIS- 110/130 immunization doses.
- FIG. 15B are bar charts illustrating expansion of CD8+ OT -I T-cells on days 7 and 17.
- FIG. 15C are bar charts illustrating expansion of CD8+ OT-I T-cells on days 19, 21 , 24, 26, 28, 33, 38, and 41 .
- 15D are bar charts illustrating expansion of CD8+ OT-I T-cells on days 45, 48, and 54. Data represent mean percent ⁇ SEM. Statistical analysis was Mann-Whitney, *p ⁇ 0.05, **p ⁇ 0.01 , ***p ⁇ 0.001 ;‘ns’ denotes p>0.05 or‘not significant’.
- FIGS. 16, 17 and 18 illustrate percent of T-cells in the peripheral blood for SLECs, MPECs, Activated/C D44 hi CD8+ endogenous and exogenous (OT-I), T-cells, on day 7 of the study of FIG. 13.
- FIG. 16 are graphs illustrating flow cytometry gating strategy for MPECs and SLECs.
- FIG. 17 are bar charts illustrating MPECs and SLEC for endogenous CD8+ T-cells.
- FIG. 18 bar charts illustrating percent of CD44 hi endogenous CD8+ T-cells (% CD8+ CD44+ T cells).
- FIGS. 19 and 20 are graphs illustrating tumor growth delay/inhibition over time, in the study of FIG. 13.
- FIG. 19 shows tumor diameter (in mm 3 ) for each of the dose ratio groups, for days 0-28, as mean ⁇ SEM grouped tumor diameter growth curves.
- FIG. 20 are bar charts (left panel) illustrating tumor weights (in grams) as mean ⁇ SEM, and scatter plots (right panel) illustrating individual tumors (in grams) as mean ⁇ SEM.
- Statistical analysis performed was Mann-Whitney, * p ⁇ 0.05, ** p ⁇ 0.01 ;‘ns’ denotes p>0.05 or‘not significant'.
- FIG. 21 are bar charts illustrating percent of CD3+ CD8+ tetramer-TRP2+ T-cells in the spleen on day 55 of the study of FIG. 13. Graphed values for gated samples are shown, and represent mean percent ⁇ SEM. Statistical analysis performed was Mann-Whitney, * p ⁇ 0.05, ** p ⁇ 0.01 ;‘ns’ denotes p>0.05 or‘not significant'.
- FIG. 22 are bar charts illustrating percent of CD3+ CD8+ eGFP/OT-1 + T-cells in the spleen and blood on day 55 of the study of FIG. 13.
- CD8+eGFP/OT-1 + T-cells gated for blood and spleen are shown, and represent mean percent ⁇ SEM.
- Statistical analysis performed was Mann-Whitney.
- FIG. 23 are bar charts illustrating splenocytes phenotypes on day 55. Data shows percent of CD3+ CD4+ PD-1 + T cells in the spleen on day 55 of the study of FIG. 13. Data represent mean percent ⁇ SEM. Statistical analysis performed was Mann-Whitney,‘ns’ denotes p>0.05 or‘not significant'.
- FIG. 24 are bar charts illustrating percent of CD3+ CD4+ CD44/CD62L central memory T-cells in the spleen on day 55 of the study of FIG. 13. Data represent mean percent ⁇ SEM. Statistical analysis performed was Mann-Whitney, * p ⁇ 0.05, ** p ⁇ 0.01.
- FIG. 25 are bar charts illustrating tumor infiltrating lymphocytes (TILs) phenotypes.
- TILs tumor infiltrating lymphocytes
- CD8+ TILs % CD8+ CD3+ T-cells
- Data represent mean percent ⁇ SEM.
- Statistical analysis performed was Mann-Whitney, * p ⁇ 0.05,‘ns’ denotes p>0.05 or‘not significant'.
- FIG. 26 are bar charts illustrating tumor infiltrating lymphocytes (TILs) phenotypes.
- TILs tumor infiltrating lymphocytes
- CD4+ TILs % CD4+ CD3+ T cells
- Data represent mean percent ⁇ SEM from.
- Statistical analysis performed was Mann-Whitney, * p ⁇ 0.05; ‘ns’ denotes p>0.05 or‘not significant'.
- the various secretable proteins i.e., vaccine proteins as described herein, can be used to stimulate an immune response in vivo.
- secretable heat-shock protein gp96-lg based allogeneic cellular vaccines can achieve high-frequency polyclonal CD8+ T cell responses to femto-molar concentrations of tumor antigens through antigen cross-priming in vivo ( Oizumi et al, J Immunol 2007, 179(4) :2310-2317) .
- Multiple immunosuppressive mechanisms elaborated by established tumors can dampen the activity of this vaccine approach, however.
- Synergistic anti-tumor benefits may result from triple combinations of gp96-lg vaccination, PD-1 blockade, and T cell costimulation using one or of an agonist of 0X40 ⁇ e.g., an 0X40 ligand-lg (OX40L-lg) fusion, or a fragment thereof that binds 0X40), an agonist of inducible T-cell costimulator (ICOS) ⁇ e.g., an ICOS ligand-lg (ICOSL-lg) fusion, or a fragment thereof that binds ICOS), an agonist of CD40 (e.g., a CD40L- Ig fusion protein, or fragment thereof), an agonist of CD27 (e.g., a CD70-lg fusion protein or fragment thereof), an agonist of 4-1 BB (e.g., a 4-1 BB ligand-lg (4-1 BBL-lg) fusion, or a fragment thereof that binds 4-1
- Vaccine proteins can induce immune responses that find use in the present invention.
- the disclosure provides a cell based therapy comprising a first cell comprising an expression vector comprising, a nucleotide sequence that encode a secretable vaccine protein and a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein.
- Compositions useful in the cell based therapy of the present invention are also provided. In various embodiments, such compositions are utilized in methods of treating subjects to stimulate immune responses in the subject including enhancing the activation of antigen-specific T cells in the subject. The present compositions find use in the treatment of various diseases including cancer.
- the heat shock protein (hsp) gp96 localized in the endoplasmic reticulum (ER), serves as a chaperone for peptides on their way to MHC class I and II molecules.
- Gp96 obtained from tumor cells and used as a vaccine can induce specific tumor immunity, presumably through the transport of tumor- specific peptides to antigen-presenting cells (APCs) (J Immunol 1999, 163(10):5178-5182).
- APCs antigen-presenting cells
- gp96-associated peptides are cross-presented to CD8 cells by dendritic cells (DCs).
- a vaccination system was developed for antitumor therapy by transfecting a gp96-lg G1-Fc fusion protein into tumor cells, resulting in secretion of gp96-lg in complex with chaperoned tumor peptides (see, J Immunother 2008, 31 (4):394-401 , and references cited therein).
- Parenteral administration of gp96-lg secreting tumor cells triggers robust, antigen-specific CD8 cytotoxic T lymphocyte (CTL) expansion, combined with activation of the innate immune system.
- CTL cytotoxic T lymphocyte
- Tumor-secreted gp96 causes the recruitment of DCs and natural killer (NK) cells to the site of gp96 secretion, and mediates DC activation.
- NK natural killer
- the cell based therapy provided herein involve a first nucleotide sequence that encodes a gp96- Ig fusion protein.
- the coding region of human gp96 is 2,412 bases in length (SEQ ID NO:1), and encodes an 803 amino acid protein (SEQ ID NO:2) that includes a 21 amino acid signal peptide at the amino terminus, a potential transmembrane region rich in hydrophobic residues, and an ER retention peptide sequence at the carboxyl terminus (GENBANK® Accession No. X15187; see, Maki et al., Proc Natl Acad Sci USA 1990, 87:5658-5562).
- the DNA and protein sequences of human gp96 follow:
- a nucleic acid encoding a gp96-lg fusion sequence can be produced using the methods described in U.S. Patent No. 8,685,384, which is incorporated herein by reference in its entirety.
- the gp96 portion of a gp96-lg fusion protein can contain all or a portion of a wild type gp96 sequence ⁇ e.g., the human sequence set forth in SEQ ID NO:2).
- a secretable gp96-lg fusion protein can include the first 799 amino acids of SEQ ID NO:2, such that it lacks the C-terminal KDEL (SEQ ID NO:3) sequence.
- the gp96 portion of the fusion protein can have an amino acid sequence that contains one or more substitutions, deletions, or additions as compared to the first 799 amino acids of the wild type gp96 sequence, such that it has at least 90% ⁇ e.g., at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity to the wild type polypeptide.
- the percent sequence identity between a particular nucleic acid or amino acid sequence and a sequence referenced by a particular sequence identification number is determined as follows. First, a nucleic acid or amino acid sequence is compared to the sequence set forth in a particular sequence identification number using the BLAST 2 Sequences (BI2seq) program from the stand-alone version of BLASTZ containing BLASTN version 2.0.14 and BLASTP version 2.0.14. This stand-alone version of BLASTZ can be obtained online at fr.com/blast or at ncbi.nlm.nih.gov. Instructions explaining how to use the BI2seq program can be found in the readme file accompanying BLASTZ.
- BI2seq BLAST 2 Sequences
- BI2seq performs a comparison between two sequences using either the BLASTN or BLASTP algorithm.
- BLASTN is used to compare nucleic acid sequences
- BLASTP is used to compare amino acid sequences.
- the options are set as follows: -i is set to a file containing the first nucleic acid sequence to be compared ⁇ e.g., C: ⁇ seq1 .bet); -j is set to a file containing the second nucleic acid sequence to be compared ⁇ e.g., C: ⁇ seq2.txt); -p is set to blastn; -o is set to any desired file name (e.g., C: ⁇ output.txt); -q is set to -1 ; -r is set to 2; and all other options are left at their default setting.
- the following command can be used to generate an output file containing a comparison between two sequences: C: ⁇ BI2seq -i c: ⁇ seq1 .txt -j c: ⁇ seq2.txt -p blastn -o c: ⁇ output.txt -q -1 -r 2.
- BI2seq are set as follows: -i is set to a file containing the first amino acid sequence to be compared (e.g., C: ⁇ seq1 .txt); -j is set to a file containing the second amino acid sequence to be compared (e.g., C: ⁇ seq2.txt); -p is set to blastp; -o is set to any desired file name (e.g., C: ⁇ output.txt); and all other options are left at their default setting.
- -i is set to a file containing the first amino acid sequence to be compared (e.g., C: ⁇ seq1 .txt)
- -j is set to a file containing the second amino acid sequence to be compared (e.g., C: ⁇ seq2.txt)
- -p is set to blastp
- -o is set to any desired file name (e.g., C: ⁇ output.txt); and all other
- the following command can be used to generate an output file containing a comparison between two amino acid sequences: C: ⁇ BI2seq -i c: ⁇ seq1.txt -j c: ⁇ seq2.txt -p blastp -o c: ⁇ output.txt. If the two compared sequences share homology, then the designated output file will present those regions of homology as aligned sequences. If the two compared sequences do not share homology, then the designated output file will not present aligned sequences.
- the number of matches is determined by counting the number of positions where an identical nucleotide or amino acid residue is presented in both sequences.
- the percent sequence identity is determined by dividing the number of matches either by the length of the sequence set forth in the identified sequence (e.g., SEQ ID NO:1 ), or by an articulated length (e.g., 100 consecutive nucleotides or amino acid residues from a sequence set forth in an identified sequence), followed by multiplying the resulting value by 100.
- the gp96 portion of nucleic acid encoding a gp96-lg fusion polypeptide can encode an amino acid sequence that differs from the wild type gp96 polypeptide at one or more amino acid positions, such that it contains one or more conservative substitutions, non conservative substitutions, splice variants, isoforms, homologues from other species, and polymorphisms.
- a "conservative substitution” denotes the replacement of an amino acid residue by another, biologically similar, residue.
- biological similarity reflects substitutions on the wild type sequence with conserved amino acids. For example, conservative amino acid substitutions would be expected to have little or no effect on biological activity, particularly if they represent less than 10% of the total number of residues in the polypeptide or protein. Conservative substitutions may be made, for instance, on the basis of similarity in polarity, charge, size, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the amino acid residues involved.
- the 20 naturally occurring amino acids can be grouped into the following six standard amino acid groups: (1) hydrophobic: Met, Ala, Val, Leu, lie; (2) neutral hydrophilic: Cys, Ser, Thr; Asn, Gin; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe. Accordingly, conservative substitutions may be effected by exchanging an amino acid by another amino acid listed within the same group of the six standard amino acid groups shown above. For example, the exchange of Asp by Glu retains one negative charge in the so modified polypeptide.
- glycine and proline may be substituted for one another based on their ability to disrupt a-helices.
- Additional examples of conserved amino acid substitutions include, without limitation, the substitution of one hydrophobic residue for another, such as isoleucine, valine, leucine, or methionine, or the substitution of one polar residue for another, such as the substitution of arginine for lysine, glutamic for aspartic acid, or glutamine for asparagine, and the like.
- conservative substitution also includes the use of a substituted amino acid residue in place of an un-substituted parent amino acid residue, provided that antibodies raised to the substituted polypeptide also immunoreact with the un-substituted polypeptide.
- non-conservative substitutions are defined as exchanges of an amino acid by another amino acid listed in a different group of the six standard amino acid groups (1) to (6) shown above.
- the substitutions may also include non-classical amino acids ⁇ e.g., selenocysteine, pyrrolysine, N-formylmethionine b-alanine, GABA and d-Aminolevulinic acid, 4- aminobenzoic acid (PABA), D-isomers of the common amino acids, 2,4-diaminobutyric acid, a-amino isobutyric acid, 4-aminobutyric acid, Abu, 2-amino butyric acid, y-Abu, e-Ahx, 6-amino hexanoic acid, Aib, 2-amino isobutyric acid, 3-amino propionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosme, citrulline, homocitrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylg
- non-classical amino acids
- Mutations may also be made to the nucleotide sequences of the present fusion proteins by reference to the genetic code, including taking into account codon degeneracy.
- the Ig portion ("tag”) of a gp96-lg fusion protein can contain, for example, a non-variable portion of an immunoglobulin molecule ⁇ e.g., an lgG1 , lgG2, lgG3, lgG4, IgM, IgA, or IgE molecule).
- immunoglobulin molecule e.g., an lgG1 , lgG2, lgG3, lgG4, IgM, IgA, or IgE molecule.
- such portions contain at least functional CH2 and CH3 domains of the constant region of an immunoglobulin heavy chain. Fusions also can be made using the carboxyl terminus of the Fc portion of a constant domain, or a region immediately amino-terminal to the CH1 of the heavy or light chain.
- the Ig tag can be from a mammalian ⁇ e.g., human, mouse, monkey, or rat) immunoglobulin, but human immunoglobulin can be particularly useful when the gp96-lg fusion is intended for in vivo use for humans.
- DNAs encoding immunoglobulin light or heavy chain constant regions are known or readily available from cDNA libraries. See, for example, Adams et al., Biochemistry 1980, 19:271 1-2719; Gough et al., Biochemistry 1980 19:2702-2710; Dolby et al., Proc Natl Acad Sci USA 1980, 77:6027-6031 ; Rice et al., Proc Natl Acad Sci USA 1982, 79:7862-7865; Falkner et al., Nature 1982, 298:286-288; and Morrison et al., Ann Rev Immunol 1984, 2:239-256.
- gp96-lg fusion proteins can readily be detected and quantified by a variety of immunological techniques known in the art, such as enzyme- linked immunosorbent assay (ELISA), immunoprecipitation, and fluorescence activated cell sorting (FACS).
- ELISA enzyme- linked immunosorbent assay
- FACS fluorescence activated cell sorting
- the peptide tag is an epitope with readily available antibodies, such reagents can be used with the techniques mentioned above to detect, quantitate, and isolate gp96-lg fusions.
- the gp96-lg fusion protein and/or the costimulatory molecule fusions comprises a linker.
- the linker may be derived from naturally-occurring multi- domain proteins or are empirical linkers as described, for example, in Chichili et al., (2013), Protein Sci. 22(2):153-167, Chen et al., (2013), Adv Drug Deliv Rev. 65(10):1357-1369, the entire contents of which are hereby incorporated by reference.
- the linker may be designed using linker designing databases and computer programs such as those described in Chen et al., (2013), Adv Drug Deliv Rev. 65(10):1357-1369 and Crasto et. al., (2000), Protein Eng. 13(5):309-312, the entire contents of which are hereby incorporated by reference.
- the linker is a synthetic linker such as PEG.
- the linker is a polypeptide. In some embodiments, the linker is less than about 100 amino acids long. For example, the linker may be less than about 100, about 95, about 90, about 85, about 80, about 75, about 70, about 65, about 60, about 55, about 50, about 45, about 40, about 35, about 30, about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 12, about 11 , about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, or about 2 amino acids long. In some embodiments, the linker is flexible. In another embodiment, the linker is rigid. In various embodiments, the linker is substantially comprised of glycine and serine residues (e.g. about 30%, or about 40%, or about 50%, or about 60%, or about 70%, or about 80%, or about 90%, or about 95%, or about 97% glycines and serines).
- glycine and serine residues e.g. about 30%, or about 40%, or about 50%, or about 60%, or
- the linker is a hinge region of an antibody ⁇ e.g., of IgG, IgA, IgD, and IgE, inclusive of subclasses (e.g. lgG1 , lgG2, lgG3, and lgG4, and lgA1 and lgA2)).
- the hinge region found in IgG, IgA, IgD, and IgE class antibodies, acts as a flexible spacer, allowing the Fab portion to move freely in space.
- the hinge domains are structurally diverse, varying in both sequence and length among immunoglobulin classes and subclasses.
- the length and flexibility of the hinge region varies among the IgG subclasses.
- the hinge region of lgG1 encompasses amino acids 216-231 and, because it is freely flexible, the Fab fragments can rotate about their axes of symmetry and move within a sphere centered at the first of two inter-heavy chain disulfide bridges.
- lgG2 has a shorter hinge than lgG1 , with 12 amino acid residues and four disulfide bridges.
- the hinge region of lgG2 lacks a glycine residue, is relatively short, and contains a rigid poly-proline double helix, stabilized by extra inter-heavy chain disulfide bridges. These properties restrict the flexibility of the lgG2 molecule.
- lgG3 differs from the other subclasses by its unique extended hinge region (about four times as long as the lgG1 hinge), containing 62 amino acids (including 21 prolines and 11 cysteines), forming an inflexible poly-proline double helix.
- the Fab fragments are relatively far away from the Fc fragment, giving the molecule a greater flexibility.
- the elongated hinge in lgG3 is also responsible for its higher molecular weight compared to the other subclasses.
- the hinge region of lgG4 is shorter than that of lgG1 and its flexibility is intermediate between that of lgG1 and lgG2. The flexibility of the hinge regions reportedly decreases in the order lgG3>lgG1 >lgG4>lgG2.
- the linker may be functional.
- the linker may function to improve the folding and/or stability, improve the expression, improve the pharmacokinetics, and/or improve the bioactivity of the present compositions.
- the linker may function to target the compositions to a particular cell type or location.
- a gp96 peptide can be fused to the hinge, CH2 and CH3 domains of murine lgG1 (Bowen et al., J Immunol 1996, 156:442-449).
- This region of the lgG1 molecule contains three cysteine residues that normally are involved in disulfide bonding with other cysteines in the Ig molecule. Since none of the cysteines are required for the peptide to function as a tag, one or more of these cysteine residues can be substituted by another amino acid residue, such as, for example, serine.
- leader sequences known in the art also can be used for efficient secretion of gp96-lg fusion proteins from bacterial and mammalian cells (see, von Heijne, J Mol Biol 1985, 184:99-105).
- Leader peptides can be selected based on the intended host cell, and may include bacterial, yeast, viral, animal, and mammalian sequences.
- the herpes virus glycoprotein D leader peptide is suitable for use in a variety of mammalian cells.
- leader peptide for use in mammalian cells can be obtained from the V-J2-C region of the mouse immunoglobulin kappa chain (Bernard et al., Proc Natl Acad Sci USA 1981 , 78:5812-5816). DNA sequences encoding peptide tags or leader peptides are known or readily available from libraries or commercial suppliers, and are suitable in the fusion proteins described herein.
- various heat shock proteins are among the vaccine proteins.
- the heat shock protein is one or more of a small hsp, hsp40, hsp60, hsp70, hsp90, and hsp110 family member, inclusive of fragments, variants, mutants, derivatives or combinations thereof (Hickey, et al., 1989, Mol. Cell. Biol. 9:2615-2626; Jindal, 1989, Mol. Cell. Biol. 9:2279-2283).
- the cell based therapy using the expression vectors provided herein can encode one or more biological response modifiers.
- the cell based therapies can encode one or more T cell costimultory molecules.
- the cell based therapies allow for a robust, antigen-specific CD8 cytotoxic T lymphocyte (CTL) expansion.
- CTL cytotoxic T lymphocyte
- the cell based therapies selectively enhance CD8 cytotoxic T lymphocyte (CTL) and do not substantially enhance T cell types that can be pro-tumor, and which include, but are not limited to, Tregs, CD4+ and/or CD8+ T cells expressing one or more checkpoint inhibitory receptors, Th2 cells and Th17 cells.
- Checkpoint inhibitory receptors refers to receptors (e.g., CTLA-4, B7-H3, B7-H4, TIM-3) expressed on immune cells that prevent or inhibit uncontrolled immune responses.
- the present cell based therapies do not substantially enhance FOXP3+ regulatory T cells.
- this selective CD8 T cell enhancement is in contrast to the non-specific T cell enhancement observed with a combination therapy of a gp-96 fusion and an antibody against a T cell costimultory molecule.
- the cell based therapies comprise an agonist of 0X40 ⁇ e.g., an 0X40 ligand-lg (OX40L-lg) fusion, or a fragment thereof that binds 0X40), an agonist of inducible T-cell costimulator (ICOS) ⁇ e.g., an ICOS ligand-lg (ICOSL-lg) fusion, or a fragment thereof that binds ICOS), an agonist of CD40 (e.g., a CD40L-lg fusion protein, or fragment thereof), an agonist of CD27 (e.g., a CD70-lg fusion protein or fragment thereof), or an agonist of 4-1 BB (e.g., a 4-1 BB ligand-lg (4-1 BBL-lg) fusion, or a fragment thereof that binds 4-1 BB).
- 0X40 e.g., an 0X40 ligand-lg (OX40L-lg)
- the cell based therapies comprise a vector which encodes an agonist of TNFRSF25 (e.g., a TL1 A-lg fusion, or a fragment thereof that binds TNFRSF25), or an agonist of glucocorticoid-induced tumor necrosis factor receptor (GITR) (e.g., a GITR ligand-lg (GITRL-lg) fusion, or a fragment thereof that binds GITR), or an agonist of CD40 (e.g., a CD40 ligand-lg (CD40L-lg) fusion, or a fragment thereof that binds CD40); or an agonist of CD27 (e.g., a CD27 ligand-lg (e.g. CD70L-lg) fusion, or a fragment thereof that binds CD40).
- GITR glucocorticoid-induced tumor necrosis factor receptor
- CD40 e.g., a CD40 ligand
- ICOS is an inducible T cell costimulatory receptor molecule that displays some homology to CD28 and CTLA-4, and interacts with B7-H2 expressed on the surface of antigen-presenting cells. ICOS has been implicated in the regulation of cell-mediated and humoral immune responses.
- 4-1 BB is a type 2 transmembrane glycoprotein belonging to the TNF superfamily, and is expressed on activated T Lymphocytes.
- 0X40 (also referred to as CD134 or TNFRSF4) is a T cell costimulatory molecule that is engaged by OX40L, and frequently is induced in antigen presenting cells and other cell types. 0X40 is known to enhance cytokine expression and survival of effector T cells.
- GITR TNFRSF18
- GITRL GITRL
- FoxP3+ regulatory T cells GITR plays a significant role in the maintenance and function of Treg within the tumor microenvironment.
- TNFRSF25 is a T cell costimulatory molecule that is preferentially expressed in CD4+ and CD8+ T cells following antigen stimulation. Signaling through TNFRSF25 is provided by TL1A, and functions to enhance T cell sensitivity to IL-2 receptor mediated proliferation in a cognate antigen dependent manner.
- CD40 is a costimulatory protein found on various antigen presenting cells which plays a role in their activation.
- the binding of CD40L (CD154) on TH cells to CD40 activates antigen presenting cells and induces a variety of downstream effects.
- CD27 a T cell costimulatory molecule belonging to the TNF superfamily which plays a role in the generation and long-term maintenance of T cell immunity. It binds to a ligand CD70 in various immunological processes.
- Additional costimulatory molecules that may be utilized in the present invention include, but are not limited to, HVEM, CD28, CD30, CD30L, CD40, CD70, LIGHT (CD258), B7-1 , and B7-2.
- the Ig portion ("tag”) of the T cell costimulatory fusion protein can contain, a non-variable portion of an immunoglobulin molecule ⁇ e.g., an lgG1 , lgG2, lgG3, lgG4, IgM, IgA, or IgE molecule). As described above, such portions typically contain at least functional CH2 and CH3 domains of the constant region of an immunoglobulin heavy chain.
- a T cell costimulatory peptide can be fused to the hinge, CH2 and CH3 domains of murine lgG1 (Bowen et al., J Immunol 1996, 156:442-449).
- the Ig tag can be from a mammalian ⁇ e.g., human, mouse, monkey, or rat) immunoglobulin, but human immunoglobulin can be particularly useful when the fusion protein is intended for in vivo use for humans.
- DNAs encoding immunoglobulin light or heavy chain constant regions are known or readily available from cDNA libraries.
- Various leader sequences as described above also can be used for secretion of T cell costimulatory fusion proteins from bacterial and mammalian cells.
- a representative nucleotide sequence (SEQ ID NO:4) encoding the extracellular domain of human ICOSL fused to Ig, and the amino acid sequence of the encoded fusion (SEQ ID NO:5) are provided:
- KSLSLSLGK (SEQ ID NO:5).
- a representative nucleotide sequence (SEQ ID NO:6) encoding the extracellular domain of human 4-1 BBL fused to Ig, and the encoded amino acid sequence (SEQ ID NO:7) are provided:
- a representative nucleotide sequence (SEQ ID NO:8) encoding the extracellular domain of human TL1 A fused to Ig, and the encoded amino acid sequence (SEQ ID NO:9) are provided:
- a representative nucleotide sequence (SEQ ID NO:10) encoding human OX40L-lg, and the encoded amino acid sequence (SEQ ID NO:11) are provided:
- HLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHY QPYAPPRDFAAYRS (SEQ ID NO:29).
- the present invention provides for variants comprising any of the sequences described herein, for instance, a sequence having at least about 60%, or at least about 61%, or at least about 62%, or at least about 63%, or at least about 64%, or at least about 65%, or at least about 66%, or at least about 67%, or at least about 68%, or at least about 69%, or at least about 70%, or at least about 71%, or at least about 72%, or at least about 73%, or at least about 74%, or at least about 75%, or at least about 76%, or at least about 77%, or at least about 78%, or at least about 79%, or at least about 80%, or at least about 81%, or at least about 82%, or at least about 83%, or at least about 84%, or at least about 85%, or at least about 86%, or at least about 87%, or at least about 88%, or at least about 89%, or at least about 90%, or at least about 91 %
- the present invention provides for an amino acid sequence having one or more amino acid mutations relative any of the protein sequences described herein.
- the one or more amino acid mutations may be independently selected from conservative or non-conservative substitutions, insertions, deletions, and truncations as described herein.
- Some human tumors can be eliminated by a patient's immune system.
- administration of a monoclonal antibody targeted to an immune "checkpoint” molecule can lead to complete response and tumor remission.
- a mode of action of such antibodies is through inhibition of an immune regulatory molecule that the tumors have co-opted as protection from an anti-tumor immune response.
- an antagonistic antibody By inhibiting these "checkpoint” molecules ⁇ e.g., with an antagonistic antibody), a patient's CD8+ T cells may be allowed to proliferate and destroy tumor cells.
- administration of a monoclonal antibody targeted to by way of example, without limitation, CTLA-4 or PD-1 can lead to complete response and tumor remission.
- the mode of action of such antibodies is through inhibition of CTLA-4 or PD-1 that the tumors have co-opted as protection from an anti-tumor immune response.
- a patient's CD8+ T cells may be allowed to proliferate and destroy tumor cells.
- the cell based therapies provided herein can be used in combination with one or more blocking antibodies targeted to an immune "checkpoint” molecule.
- the cell based therapies provided herein can be used in combination with one or more blocking antibodies targeted to a molecule such as CTLA-4 or PD-1.
- the cell based therapies provided herein may be used in combination with an agent that blocks, reduces and/or inhibits PD-1 and PD-L1 or PD- L2 and/or the binding of PD-1 with PD-L1 or PD-L2 (by way of non-limiting example, one or more of nivolumab (ONO-4538/BMS-936558, MDX1106, OPDIVO, BRISTOL MYERS SQUIBB), pembrolizumab (KEYTRUDA, Merck), pidilizumab (CT-011 , CURE TECH), MK-3475 (MERCK), BMS 936559 (BRISTOL MYERS SQUIBB), MPDL3280A (ROCHE)).
- an agent that blocks, reduces and/or inhibits PD-1 and PD-L1 or PD- L2 and/or the binding of PD-1 with PD-L1 or PD-L2
- the cell based therapies provided herein may be used in combination with an agent that blocks, reduces and/or inhibits the activity of CTLA-4 and/or the binding of CTLA-4 with one or more receptors (e.g. CD80, CD86, AP2M1 , SHP-2, and PPP2R5A).
- the immune-modulating agent is an antibody such as, by way of non-limitation, ipilimumab (MDX-010, M DX- 101 , Yervoy, BMS) and/or tremelimumab (Pfizer). Blocking antibodies against these molecules can be obtained from, for example, Bristol Myers Squibb (New York, NY), Merck (Kenilworth, NJ), Medlmmune (Gaithersburg, MD), and Pfizer (New York, NY).
- the cell based therapies provided herein can be used in combination with one or more blocking antibodies targeted to an immune "checkpoint” molecule such as for example, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160 (also referred to as BY55), CGEN-15049, CHK 1 and CHK2 kinases, A2aR, CEACAM ⁇ e.g., CEACAM-1 , CEACAM-3 and/or CEACAM-5), GITR, GITRL, galectin-9, CD244, CD160, TIGIT, SIRPa, ICOS, CD172a, and TMIGD2 and various B-7 family ligands (including, but are not limited to, B7-1 , B7-2, B7-DC, B7-H1 , B7-H2, B7-H3, B7-H4, B7-H5, B7-H6 and B7-H7).
- Cell based therapies including, but are not limited to, B7-1
- the present disclosure provides a cell based therapy comprising a first cell containing an expression vector comprising, a nucleotide sequence that encode a secretable vaccine protein ⁇ e.g., a gp96-lg fusion protein) and a second cell containing an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein, ⁇ e.g., OX40L-lg or a portion thereof that binds specifically to 0X40, ICOSL-lg or a portion thereof that binds specifically to ICOS, 4-1 BBL-lg, or a portion thereof that binds specifically to 4-1 BBR, CD40L-lg, or a portion thereof that binds specifically to CD40, CD70-lg, or a portion thereof that binds specifically to CD27, TL1 A-lg or a portion thereof that binds specifically to TNFRSF25, or GITRL-lg or a portion thereof that binds
- present disclosure provides methods for making the cell based therapies described herein, as well as methods for administering the cell based therapies.
- the methods provided herein include administering to a patient an effective amount of a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein, wherein the patient is undergoing a treatment with a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject.
- the methods provided herein include administering to a patient an effective amount of a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein, wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject, and wherein the patient is undergoing a treatment with a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein.
- the methods provided herein include administering to the patient an effective amount of a first cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject.
- gp96-lg based vaccines can be generated to stimulate antigen specific immune responses against individual antigens expressed by simian immunodeficiency virus, human immunodeficiency virus, hepatitis C virus and malaria. Immune responses to these vaccines may be enhanced through a cell based therapy of a T cell costimulatory fusion protein and a cell based therapy of gp96-lg.
- cDNA or DNA sequences encoding a vaccine protein fusion e.g., a gp96-lg fusion
- a T cell costimulatory fusion protein can be obtained (and, if desired, modified) using conventional DNA cloning and mutagenesis methods, DNA amplification methods, and/or synthetic methods.
- a sequence encoding a vaccine protein fusion protein ⁇ e.g., a gp96-lg fusion protein) and/or a T cell costimulatory fusion protein can be inserted into a cloning vector for genetic modification and replication purposes prior to expression.
- Each coding sequence can be operably linked to a regulatory element, such as a promoter, for purposes of expressing the encoded protein in suitable host cells in vitro and in vivo.
- the cell based therapy can be administered to produce secreted vaccine proteins (e.g., gp96- Ig) and T cell costimulatory fusion proteins.
- Cells may be cultured in vitro or genetically engineered, for example.
- Host cells can be obtained from normal or affected subjects, including healthy humans, cancer patients, and patients with an infectious disease, private laboratory deposits, public culture collections such as the American Type Culture Collection, or from commercial suppliers.
- Cells that can be used for production and secretion of gp96-lg fusion proteins and T cell costimulatory fusion proteins in vivo include, without limitation, epithelial cells, endothelial cells, keratinocytes, fibroblasts, muscle cells, hepatocytes; blood cells such as T lymphocytes, B lymphocytes, monocytes, macrophages, neutrophils, eosinophils, megakaryocytes, or granulocytes, various stem or progenitor cells, such as hematopoietic stem or progenitor cells (e.g., as obtained from bone marrow), umbilical cord blood, peripheral blood, fetal liver, etc., and tumor cells (e.g., human tumor cells).
- the choice of cell type depends on the type of tumor or infectious disease being treated or prevented, and can be determined by one of skill in the art.
- Different host cells have characteristic and specific mechanisms for post-translational processing and modification of proteins.
- a host cell may be chosen which modifies and processes the expressed gene products in a specific fashion similar to the way the recipient processes its heat shock proteins (hsps).
- hsps heat shock proteins
- the type of host cell has been used for expression of heterologous genes, and is reasonably well characterized and developed for large-scale production processes.
- the host cells are autologous to the patient to whom the present fusion or recombinant cells secreting the present fusion proteins are subsequently administered.
- the cell based therapy as provided herein can be introduced into an antigenic cell.
- antigenic cells can include preneoplastic cells that are infected with a cancer-causing infectious agent, such as a virus, but that are not yet neoplastic, or antigenic cells that have been exposed to a mutagen or cancer-causing agent, such as a DNA-damaging agent or radiation, for example.
- a cancer-causing infectious agent such as a virus
- antigenic cells that have been exposed to a mutagen or cancer-causing agent, such as a DNA-damaging agent or radiation, for example.
- Other cells that can be used are preneoplastic cells that are in transition from a normal to a neoplastic form as characterized by morphology or physiological or biochemical function.
- the cancer cells and preneoplastic cells used in the methods provided herein are of mammalian origin. Mammals contemplated include humans, companion animals ⁇ e.g., dogs and cats), livestock animals ⁇ e.g., sheep, cattle, goats, pigs and horses), laboratory animals (e.g., mice, rats and rabbits), and captive or free wild animals.
- cancer cells e.g., human tumor cells
- the cancer cells provide antigenic peptides that become associated non-covalently with the expressed gp96-lg fusion proteins.
- Cell lines derived from a preneoplastic lesion, cancer tissue, or cancer cells also can be used, provided that the cells of the cell line have at least one or more antigenic determinant in common with the antigens on the target cancer cells.
- Cancer tissues, cancer cells, cells infected with a cancer-causing agent, other preneoplastic cells, and cell lines of human origin can be used. Cancer cells excised from the patient to whom ultimately the fusion proteins ultimately are to be administered can be particularly useful, although allogeneic cells also can be used.
- a cancer cell can be from an established tumor cell line such as, without limitation, an established non-small cell lung carcinoma (NSCLC), bladder cancer, melanoma, ovarian cancer, renal cell carcinoma, prostate carcinoma, sarcoma, breast carcinoma, squamous cell carcinoma, head and neck carcinoma, hepatocellular carcinoma, pancreatic carcinoma, or colon carcinoma cell line.
- NSCLC non-small cell lung carcinoma
- bladder cancer bladder cancer
- melanoma ovarian cancer
- renal cell carcinoma prostate carcinoma
- sarcoma breast carcinoma
- squamous cell carcinoma head and neck carcinoma
- pancreatic carcinoma pancreatic carcinoma
- the present fusion proteins allow for both the costimulation T cell and the presentation of various tumor cell antigens.
- the present vaccine protein fusions e.g., gp96 fusions
- the tumor cells secrete a variety of antigens.
- antigens that can be secreted are: ACRBP, ACTL8, ADAM2, ADAM29, AKAP3, AKAP4, ANKRD45, ARMC3, ARX, ATAD2, BAGE, BAGE2, BAGE3, BAGE4, BAGE5, BRDT, C15ORF60, C210RF99, CABYR, CAGE1 , CALR3, CASC5, CCDC110, CCDC33, CCDC36, CCDC62, CCDC83, CDCA1 , CEP290, CEP55, COX6B2, CPXCR1 , CRISP2, CSAG1 , CSAG2, CSAG3B, CT16.2, CT45A1 , CT45A2, CT45A3, CT45A4, CT45A5, CT45A6, CT47A1 , CT47A10, CT47A11 , CT47A2, CT47A3, CT47A4, CT47A5, CT47A6, CT47A7, CT47A8, CT47A9, CT47B1
- the antigens are human endogenous retroviral antigens.
- Illustrative antigens can also include antigens from human endogenous retroviruses which include, but are not limited to, epitopes derived from at least a portion of Gag, at least a portion of T at, at least a portion of Rev, a least a portion of Nef, and at least a portion of gp160.
- the present vaccine protein fusions ⁇ e.g., gp96 fusions
- the present vaccine protein fusions provide for an adjuvant effect that further allows the immune system of a patient, when used in the various methods described herein, to be activated against a disease of interest.
- Both prokaryotic and eukaryotic vectors can be used for expression of the vaccine protein (e.g., gp96-lg) and T cell costimulatory fusion proteins in the cell based therapy methods provided herein.
- Prokaryotic vectors include constructs based on E. coli sequences (see, e.g., Makrides, Microbiol Rev 1996, 60:512-538). Non-limiting examples of regulatory regions that can be used for expression in E.
- prokaryotic expression vectors may include the Agt vector series such as Agt11 (Huynh et al., in "DNA Cloning Techniques, Vol. I: A Practical Approach,” 1984, (D. Glover, ed.), pp. 49-78, IRL Press, Oxford), and the pET vector series (Studier et al., Methods Enzymol 1990, 185:60-89).
- Agt vector series such as Agt11 (Huynh et al., in "DNA Cloning Techniques, Vol. I: A Practical Approach,” 1984, (D. Glover, ed.), pp. 49-78, IRL Press, Oxford)
- pET vector series Studdier et al., Methods Enzymol 1990, 185:60-89.
- Prokaryotic host-vector systems cannot perform much of the post-translational processing of mammalian cells, however. Thus, eukaryotic host- vector systems may be particularly useful.
- a variety of regulatory regions can be used for expression of the vaccine protein ⁇ e.g., gp96- Ig) and T cell costimulatory fusions in mammalian host cells.
- the SV40 early and late promoters the cytomegalovirus (CMV) immediate early promoter, and the Rous sarcoma virus long terminal repeat (RSV-LTR) promoter can be used.
- CMV cytomegalovirus
- RSV-LTR Rous sarcoma virus long terminal repeat
- Inducible promoters that may be useful in mammalian cells include, without limitation, promoters associated with the metallothionein II gene, mouse mammary tumor virus glucocorticoid responsive long terminal repeats (MMTV-LTR), the b-interferon gene, and the hsp70 gene (see, Williams et al., Cancer Res 1989, 49:2735-42; and Taylor et al., Mol Cell Biol 1990, 10:165-75). Heat shock promoters or stress promoters also may be advantageous for driving expression of the fusion proteins in recombinant host cells.
- promoters associated with the metallothionein II gene mouse mammary tumor virus glucocorticoid responsive long terminal repeats (MMTV-LTR), the b-interferon gene, and the hsp70 gene (see, Williams et al., Cancer Res 1989, 49:2735-42; and Taylor et al., Mol Cell Biol 1990, 10:165-75).
- the present invention contemplates the use of inducible promoters capable of effecting high level of expression transiently in response to a cue.
- Illustrative inducible expression control regions include those comprising an inducible promoter that is stimulated with a cue such as a small molecule chemical compound. Particular examples can be found, for example, in U.S. Pat. Nos. 5,989,910, 5,935,934, 6,015,709, and 6,004,941 , each of which is incorporated herein by reference in its entirety.
- Animal regulatory regions that exhibit tissue specificity and have been utilized in transgenic animals also can be used in tumor cells of a particular tissue type: the elastase I gene control region that is active in pancreatic acinar cells (Swift et al., Cell 1984, 38:639-646; Ornitz et al., Cold Spring Harbor Symp Quant Biol 1986, 50:399-409; and MacDonald, Hepatology 1987, 7:425-515); the insulin gene control region that is active in pancreatic beta cells (Hanahan, Nature 1985, 315:115-122), the immunoglobulin gene control region that is active in lymphoid cells (Grosschedl et al., Cell 1984, 38:647- 658; Adames et al., Nature 1985, 318:533-538; and Alexander et al., Mol Cell Biol 1987, 7:1436-1444), the mouse mammary tumor virus control region that is active in testicular, breast, lymphoid
- An expression vector also can include transcription enhancer elements, such as those found in SV40 virus, Hepatitis B virus, cytomegalovirus, immunoglobulin genes, metallothionein, and b-actin (see, Bittner et al., Meth Enzymol 1987, 153:516-544; and Gorman, Curr Op Biotechnol 1990, 1 :36-47).
- an expression vector can contain sequences that permit maintenance and replication of the vector in more than one type of host cell, or integration of the vector into the host chromosome. Such sequences include, without limitation, to replication origins, autonomously replicating sequences (ARS), centromere DNA, and telomere DNA.
- ARS autonomously replicating sequences
- an expression vector can contain one or more selectable or screenable marker genes for initially isolating, identifying, or tracking host cells that contain DNA encoding fusion proteins as described herein.
- selectable or screenable marker genes for initially isolating, identifying, or tracking host cells that contain DNA encoding fusion proteins as described herein.
- stable expression in mammalian cells can be useful.
- a number of selection systems can be used for mammalian cells.
- the Herpes simplex virus thymidine kinase (Wigler et al., Cell 1977, 11 :223), hypoxanthine-guanine phosphoribosyltransferase (Szybalski and Szybalski, Proc Natl Acad Sci USA 1962, 48:2026), and adenine phosphoribosyltransferase (Lowy et al., Cell 1980, 22:817) genes can be employed in tk-, hgprt-, or aprt- cells, respectively.
- antimetabolite resistance can be used as the basis of selection for dihydrofolate reductase (dhfr), which confers resistance to methotrexate (Wigler et al., Proc Natl Acad Sci USA 1980, 77:3567; O'Hare et al., Proc Natl Acad Sci USA 1981 , 78:1527); gpt, which confers resistance to mycophenolic acid (Mulligan and Berg, Proc Natl Acad Sci USA 1981 , 78:2072); neomycin phosphotransferase (neo), which confers resistance to the aminoglycoside G-418 (Colberre-Garapin et al., J Mol Biol 1981 , 150:1); and hygromycin phosphotransferase (hyg), which confers resistance to hygromycin (Santerre et al., Gene 1984, 30:147).
- Other selectable markers such as histidinol and Ze
- Useful mammalian host cells include, without limitation, cells derived from humans, monkeys, and rodents (see, for example, Kriegler in "Gene Transfer and Expression: A Laboratory Manual,” 1990, New York, Freeman & Co.). These include monkey kidney cell lines transformed by SV40 ⁇ e.g., COS-7, AT CC CRL 1651 ); human embryonic kidney lines (e.g., 293, 293-EBNA, or 293 cells subcloned for growth in suspension culture, Graham et al., J Gen Virol 1977, 36:59); baby hamster kidney cells (e.g., BHK, ATCC CCL 10); Chinese hamster ovary-cells-DHFR (e.g., CHO, Urlaub and Chasin, Proc Natl Acad Sci USA 1980, 77:4216); mouse sertoli cells (Mather, Biol Reprod 1980, 23:243-251 ); mouse fibroblast cells (e.g., NIH-3T3), monkey kidney cells (
- Illustrative cancer cell types for expressing the fusion proteins described herein include mouse fibroblast cell line, NIH3T3, mouse Lewis lung carcinoma cell line, LLC, mouse mastocytoma cell line, P815, mouse lymphoma cell line, EL4 and its ovalbumin transfectant, E.G7, mouse melanoma cell line, B16F10, mouse fibrosarcoma cell line, MC57, human small cell lung carcinoma cell lines, SCLC#2 and SCLC#7, human lung adenocarcinoma cell line, e.g., AD100, and human prostate cancer cell line, e.g., PC-3.
- mouse fibroblast cell line NIH3T3, mouse Lewis lung carcinoma cell line, LLC, mouse mastocytoma cell line, P815, mouse lymphoma cell line, EL4 and its ovalbumin transfectant, E.G7, mouse melanoma cell line, B16F10, mouse fibrosarcoma cell line, MC57, human small cell lung carcinoma
- a number of viral-based expression systems also can be used with mammalian cells to produce gp96-lg and T cell costimulatory fusion proteins.
- Vectors using DNA virus backbones have been derived from simian virus 40 (SV40) (Flamer et al., Cell 1979, 17:725), adenovirus (Van Doren et al., Mol Cell Biol 1984, 4:1653), adeno-associated virus (McLaughlin et al., J Virol 1988, 62:1963), and bovine papillomas virus (Zinn et al., Proc Natl Acad Sci USA 1982, 79:4897).
- SV40 simian virus 40
- adenovirus Van Doren et al., Mol Cell Biol 1984, 4:1653
- adeno-associated virus McLaughlin et al., J Virol 1988, 62:1963
- bovine papillomas virus Zainn e
- the donor DNA sequence may be ligated to an adenovirus transcription/translation control complex, e.g., the late promoter and tripartite leader sequence.
- This fusion gene may then be inserted in the adenovirus genome by in vitro or in vivo recombination. Insertion in a non-essential region of the viral genome (e.g., region E1 or E3) can result in a recombinant virus that is viable and capable of expressing heterologous products in infected hosts. (See, e.g., Logan and Shenk, Proc Natl Acad Sci USA 1984, 81 :3655-3659).
- Bovine papillomavirus can infect many higher vertebrates, including man, and its DNA replicates as an episome.
- a number of shuttle vectors have been developed for recombinant gene expression which exist as stable, multicopy (20-300 copies/cell) extrachromosomal elements in mammalian cells.
- these vectors typically contain a segment of BPV DNA (the entire genome or a 69% transforming fragment), a promoter with a broad host range, a polyadenylation signal, splice signals, a selectable marker, and "poisonless” plasmid sequences that allow the vector to be propagated in E. coli.
- the expression gene constructs are transfected into cultured mammalian cells by, for example, calcium phosphate coprecipitation.
- a dominant selectable marker such as histidinol and G418 resistance.
- the vaccinia 7.5K promoter can be used.
- vaccinia 7.5K promoter See, e.g., Mackett et al., Proc Natl Acad Sci USA 1982, 79:7415-7419; Mackett et al., J Virol 1984, 49:857-864; and Panicali et al., Proc Natl Acad Sci USA 1982, 79:4927-4931.
- vectors based on the Epstein- Barr virus (EBV) origin (OriP) and EBV nuclear antigen 1 (EBNA-1 ; a trans-acting replication factor) can be used.
- Such vectors can be used with a broad range of human host cells, e.g., EBO-pCD (Spickofsky et al., DNA Prot Eng Tech 1990, 2:14-18); pDR2 and ADR2 (available from Clontech Laboratories).
- EBO-pCD Spickofsky et al., DNA Prot Eng Tech 1990, 2:14-18
- pDR2 and ADR2 available from Clontech Laboratories.
- Gp96-lg and T cell costimulatory fusion proteins also can be made with retrovirus-based expression systems.
- Retroviruses such as Moloney murine leukemia virus, can be used since most of the viral gene sequence can be removed and replaced with exogenous coding sequence while the missing viral functions can be supplied in trans. In contrast to transfection, retroviruses can efficiently infect and transfer genes to a wide range of cell types including, for example, primary hematopoietic cells.
- the host range for infection by a retroviral vector can be manipulated by the choice of envelope used for vector packaging.
- a retroviral vector can comprise a 5' long terminal repeat (LTR), a 3' LTR, a packaging signal, a bacterial origin of replication, and a selectable marker.
- the gp96-lg fusion protein coding sequence for example, can be inserted into a position between the 5' LTR and 3' LTR, such that transcription from the 5' LTR promoter transcribes the cloned DNA.
- the 5' LTR contains a promoter ⁇ e.g., an LTR promoter), an R region, a U5 region, and a primer binding site, in that order. Nucleotide sequences of these LTR elements are well known in the art.
- a heterologous promoter as well as multiple drug selection markers also can be included in the expression vector to facilitate selection of infected cells. See, McLauchlin et al., Prog Nucleic Acid Res Mol Biol 1990, 38:91-135; Morgenstern et al., Nucleic Acid Res 1990, 18:3587-3596; Choulika et al., J Virol 1996, 70:1792-1798; Boesen et al., Biotherapy 1994, 6:291-302; Salmons and Gunzberg, Pluman Gene Ther 1993, 4:129-141 ; and Grossman and Wilson, Curr Opin Genet Devel 1993, 3:110-114.
- any of the cloning and expression vectors described herein may be synthesized and assembled from known DNA sequences using techniques that are known in the art.
- the regulatory regions and enhancer elements can be of a variety of origins, both natural and synthetic.
- Some vectors and host cells may be obtained commercially. Non-limiting examples of useful vectors are described in Appendix 5 of Current Protocols in Molecular Biology, 1988, ed. Ausubel et al., Greene Publish. Assoc. & Wiley Interscience, which is incorporated herein by reference; and the catalogs of commercial suppliers such as Clontech Laboratories, Stratagene Inc., and Invitrogen, Inc.
- Cell based therapies can be used in administration to a subject ⁇ e.g., a research animal or a mammal, such as a human, having a clinical condition such as cancer or an infection).
- the cell based therapy can be administered to a subject for the treatment of cancer or infection.
- the infection can be, for example, an acute infection or a chronic infection.
- the infection can be an infection by hepatitis C virus, hepatitis B virus, human immunodeficiency virus, or malaria.
- the methods can include administering to a subject the cell based therapies under conditions wherein the progression or a symptom of the clinical condition in the subject is reduced in a therapeutic manner.
- the present invention pertains to cancers and/or tumors; for example, the treatment or prevention of cancers and/or tumors.
- Cancers or tumors refer to an uncontrolled growth of cells and/or abnormal increased cell survival and/or inhibition of apoptosis which interferes with the normal functioning of the bodily organs and systems. Included are benign and malignant cancers, polyps, hyperplasia, as well as dormant tumors or micrometastases. Also, included are cells having abnormal proliferation that is not impeded by the immune system ⁇ e.g., virus infected cells).
- the cancer may be a primary cancer or a metastatic cancer.
- the primary cancer may be an area of cancer cells at an originating site that becomes clinically detectable, and may be a primary tumor.
- the metastatic cancer may be the spread of a disease from one organ or part to another non-adjacent organ or part.
- the metastatic cancer may be caused by a cancer cell that acquires the ability to penetrate and infiltrate surrounding normal tissues in a local area, forming a new tumor, which may be a local metastasis.
- the cancer may also be caused by a cancer cell that acquires the ability to penetrate the walls of lymphatic and/or blood vessels, after which the cancer cell is able to circulate through the bloodstream (thereby being a circulating tumor cell) to other sites and tissues in the body.
- the cancer may be due to a process such as lymphatic or hematogeneous spread.
- the cancer may also be caused by a tumor cell that comes to rest at another site, re-penetrates through the vessel or walls, continues to multiply, and eventually forms another clinically detectable tumor.
- the cancer may be this new tumor, which may be a metastatic (or secondary) tumor.
- the cancer may be caused by tumor cells that have metastasized, which may be a secondary or metastatic tumor.
- the cells of the tumor may be like those in the original tumor.
- the secondary tumor while present in the liver, is made up of abnormal breast or colon cells, not of abnormal liver cells.
- the tumor in the liver may thus be a metastatic breast cancer or a metastatic colon cancer, not liver cancer.
- the cancer may have an origin from any tissue.
- the cancer may originate from melanoma, colon, breast, or prostate, and thus may be made up of cells that were originally skin, colon, breast, or prostate, respectively.
- the cancer may also be a hematological malignancy, which may be lymphoma.
- the cancer may invade a tissue such as liver, lung, bladder, or intestinal.
- Illustrative cancers that may be treated include, but are not limited to, carcinomas, e.g. various subtypes, including, for example, adenocarcinoma, basal cell carcinoma, squamous cell carcinoma, and transitional cell carcinoma), sarcomas (including, for example, bone and soft tissue), leukemias (including, for example, acute myeloid, acute lymphoblastic, chronic myeloid, chronic lymphocytic, and hairy cell), lymphomas and myelomas (including, for example, Hodgkin and non-Hodgkin lymphomas, light chain, non-secretory, MGUS, and plasmacytomas), and central nervous system cancers (including, for example, brain (e.g.
- gliomas ⁇ e.g., astrocytoma, oligodendroglioma, and ependymoma
- meningioma astrocytoma, oligodendroglioma, and ependymoma
- pituitary adenoma a neurotrophic factor
- neuromas a spinal cord tumors (e.g., meningiomas and neurofibroma).
- Representative cancers and/or tumors of the present invention include, but are not limited to, a basal cell carcinoma, biliary tract cancer; bladder cancer; bone cancer; brain and central nervous system cancer; breast cancer; cancer of the peritoneum; cervical cancer; choriocarcinoma; colon and rectum cancer; connective tissue cancer; cancer of the digestive system; endometrial cancer; esophageal cancer; eye cancer; cancer of the head and neck; gastric cancer (including gastrointestinal cancer); glioblastoma; hepatic carcinoma; hepatoma; intra-epithelial neoplasm; kidney or renal cancer; larynx cancer; leukemia; liver cancer; lung cancer (e.g., small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous carcinoma of the lung); melanoma; myeloma; neuroblastoma; oral cavity cancer (lip, tongue, mouth, and pharynx);
- the present fusions are used to eliminate intracellular pathogens.
- the present fusions are used to treat one or more infections.
- the present fusion proteins are used in methods of treating viral infections (including, for example, HIV and HCV), parasitic infections (including, for example, malaria), and bacterial infections.
- the infections induce immunosuppression.
- HIV infections often result in immunosuppression in the infected subjects.
- the treatment of such infections may involve, in various embodiments, modulating the immune system with the present fusion proteins to favor immune stimulation over immune inhibition.
- the present invention provides methods for treating infections that induce immunoactivation.
- intestinal helminth infections have been associated with chronic immune activation.
- the treatment of such infections may involve modulating the immune system with the present fusion proteins to favor immune inhibition over immune stimulation.
- the present invention provides methods of treating viral infections including, without limitation, acute or chronic viral infections, for example, of the respiratory tract, of papilloma virus infections, of herpes simplex virus (HSV) infection, of human immunodeficiency virus (HIV) infection, and of viral infection of internal organs such as infection with hepatitis viruses.
- the viral infection is caused by a virus of family Flaviviridae.
- the virus of family Flaviviridae is selected from Yellow Fever Virus, West Nile virus, Dengue virus, Japanese Encephalitis Virus, St. Louis Encephalitis Virus, and Hepatitis C Virus.
- the viral infection is caused by a virus of family Picornaviridae, e.g., poliovirus, rhinovirus, coxsackievirus.
- the viral infection is caused by a member of Orthomyxoviridae, e.g., an influenza virus.
- the viral infection is caused by a member of Retroviridae, e.g., a lentivirus.
- the viral infection is caused by a member of Paramyxoviridae, e.g., respiratory syncytial virus, a human parainfluenza virus, rubulavirus ⁇ e.g., mumps virus), measles virus, and human metapneumovirus.
- the viral infection is caused by a member of Bunyaviridae, e.g., hantavirus. In other embodiments, the viral infection is caused by a member of Reoviridae, e.g., a rotavirus.
- the present invention provides methods of treating parasitic infections such as protozoan or helminths infections.
- the parasitic infection is by a protozoan parasite.
- the oritiziab parasite is selected from intestinal protozoa, tissue protozoa, or blood protozoa.
- Illustrative protozoan parasites include, but are not limited to, Entamoeba hystolytica, Giardia lamblia, Cryptosporidium muris, Trypanosomatida gambiense, Trypanosomatida rhodesiense, Trypanosomatida crusi, Leishmania mexicana, Leishmania braziliensis, Leishmania tropica, Leishmania donovani, Toxoplasma gondii, Plasmodium vivax, Plasmodium ovale, Plasmodium malariae, Plasmodium falciparum, Trichomonas vaginalis, and Histomonas meleagridis.
- the parasitic infection is by a helminthic parasite such as nematodes ⁇ e.g., Adenophorea).
- the parasite is selected from Secementea ⁇ e.g., Trichuris trichiura, Ascaris lumbricoides, Enterobius vermicularis, Ancylostoma duodenale, Necator americanus, Strongyloides stercoralis, Wuchereria bancrofti, Dracunculus medinensis).
- the parasite is selected from trematodes (e.g. blood flukes, liver flukes, intestinal flukes, and lung flukes).
- the parasite is selected from: Schistosoma mansoni, Schistosoma haematobium, Schistosoma japonicum, Fasciola hepatica, Fasciola gigantica, Heterophyes heterophyes, Paragonimus westermani.
- the parasite is selected from cestodes (e.g., Taenia solium, Taenia saginata, Hymenolepis nana, Echinococcus granulosus).
- the present invention provides methods of treating bacterial infections.
- the bacterial infection is by a gram-positive bacterium, gram-negative bacteria, aerobic and/or anaerobic bacteria.
- the bacteria is selected from, but not limited to, Staphylococcus, Lactobacillus, Streptococcus, Sarcina, Escherichia, Enterobacter, Klebsiella, Pseudomonas, Acinetobacter, Mycobacterium, Proteus, Campylobacter, Citrobacter, Nisseria, Baccillus, Bacteroides, Peptococcus, Clostridium, Salmonella, Shigella, Serratia, Haemophilus, Brucella and other organisms.
- the bacteria is selected from, but not limited to, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas acidovorans, Pseudomonas alcaligenes, Pseudomonas putida, Stenotrophomonas maltophilia, Burkholderia cepacia, Aeromonas hydrophilia, Escherichia coli, Citrobacter freundii, Salmonella typhimurium, Salmonella typhi, Salmonella paratyphi, Salmonella enteritidis, Shigella dysenteriae, Shigella flexneri, Shigella sonnei, Enterobacter cloacae, Enterobacter aerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Serratia marcescens, Francisella tularensis, Morganella morganii, Proteus mirabili
- the cell based therapy to be administered can be admixed, encapsulated, conjugated or otherwise associated with other molecules, molecular structures, or mixtures of compounds such as, for example, liposomes, receptor or cell targeted molecules, or oral, topical or other formulations for assisting in uptake, distribution and/or absorption.
- the cell based therapy to be administered can be in combination with a pharmaceutically acceptable carrier.
- compositions containing cell based therapies described herein, in combination with a physiologically and pharmaceutically acceptable carrier can be include any of the well-known components useful for immunization.
- the carrier can facilitate or enhance an immune response to an antigen administered in a vaccine.
- the cell formulations can contain buffers to maintain a preferred pH range, salts or other components that present an antigen to an individual in a composition that stimulates an immune response to the antigen.
- the physiologically acceptable carrier also can contain one or more adjuvants that enhance the immune response to an antigen.
- Pharmaceutically acceptable carriers include, for example, pharmaceutically acceptable solvents, suspending agents, or any other pharmacologically inert vehicles for delivering compounds to a subject.
- Pharmaceutically acceptable carriers can be liquid or solid, and can be selected with the planned manner of administration in mind so as to provide for the desired bulk, consistency, and other pertinent transport and chemical properties, when combined with one or more therapeutic compounds and any other components of a given pharmaceutical composition.
- Typical pharmaceutically acceptable carriers include, without limitation: water, saline solution, binding agents ⁇ e.g., polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers ⁇ e.g., lactose or dextrose and other sugars, gelatin, or calcium sulfate), lubricants ⁇ e.g., starch, polyethylene glycol, or sodium acetate), disintegrates (e.g., starch or sodium starch glycolate), and wetting agents (e.g., sodium lauryl sulfate).
- binding agents ⁇ e.g., polyvinylpyrrolidone or hydroxypropyl methylcellulose
- fillers ⁇ e.g., lactose or dextrose
- compositions can be formulated for subcutaneous, intramuscular, or intradermal administration, or in any manner acceptable for immunization.
- An adjuvant refers to a substance which, when added to an immunogenic agent such as a tumor cell expressing secreted vaccine protein ⁇ e.g., gp96-lg) and T cell costimulatory fusion polypeptides, nonspecifically enhances or potentiates an immune response to the agent in the recipient host upon exposure to the mixture.
- Adjuvants can include, for example, oil-in-water emulsions, water-in oil emulsions, alum (aluminum salts), liposomes and microparticles, such as, polysytrene, starch, polyphosphazene and polylactide/polyglycosides.
- Adjuvants can also include, for example, squalene mixtures (SAF-I), muramyl peptide, saponin derivatives, mycobacterium cell wall preparations, monophosphoryl lipid A, mycolic acid derivatives, nonionic block copolymer surfactants, Quil A, cholera toxin B subunit, polyphosphazene and derivatives, and immunostimulating complexes (ISCOMs) such as those described by Takahashi et al., Nature 1990, 344:873-875.
- SAF-I squalene mixtures
- IFA Incomplete Freund's Adjuvant
- Various appropriate adjuvants are well known in the art (see, for example, Warren and Chedid, CRC Critical Reviews in Immunology 1988, 8:83; and Allison and Byars, in Vaccines: New Approaches to Immunological Problems, 1992, Ellis, ed., Butterworth-Fleinemann, Boston).
- Additional adjuvants include, for example, bacille Calmett-Guerin (BCG), DETOX (containing cell wall skeleton of Mycobacterium phlei (CWS) and monophosphoryl lipid A from Salmonella minnesota (MPL)), and the like (see, for example, Floover et al., J Clin Oncol 1993, 1 1 :390; and Woodlock et al., J Immunother 1999, 22:251 -259).
- BCG Bacille Calmett-Guerin
- DETOX containing cell wall skeleton of Mycobacterium phlei
- MPL monophosphoryl lipid A from Salmonella minnesota
- a cell secreting a T cell costimulatory fusion protein ⁇ e.g., OX40L-lg can be administered to a subject at about 100,000 cells, about 150,000 cells, about 200,000 cells, about 250,000 cells, about 300,000 cells, about 350,000 cells, about 400,000 cells, about 450,000 cells, about
- a cell secreting a vaccine protein (e.g., gp96-lg) can be administered to a subject at about 100,000 cells, about 150,000 cells, about 200,000 cells, about 250,000 cells, about
- a fixed dose of a cell secreting a vaccine protein is 1 x 10 7 cells.
- a fixed dose of a cell secreting a T cell costimulatory fusion protein ⁇ e.g., OX40L-lg) is 1 x 10 7 cells.
- a cell secreting a T cell costimulatory fusion protein ⁇ e.g., OX40L- Ig) can be administered to a subject at about 0.1 -1.1 x 10 7 cells, about 0.2- 1 .1 x 10 7 cells, about 0.3- 1.1 x 10 7 cells, about 0.4- 1 .1 x 10 7 cells, about 0.5- 1 .1 x 10 7 cells, about 0.6- 1.1 x 10 7 cells, about 0.7- 1 .1 x 10 7 cells, about 0.8- 1.1 x 10 7 cells, about or 0.9- 1 .1 x 10 7 cells, about 0.1-2.1 x 10 7 cells, about 0.2-
- 6.1 x 10 7 cells about 0.3- 6.1 x 10 7 cells, about 0.4- 6.1 x 10 7 cells, about 0.5- 6.1 x 10 7 cells, about 0.6-
- 6.1 x 10 7 cells about 0.7- 6.1 x 10 7 cells, about 0.8- 6.1 x 10 7 cells, about 0.9- 6.1 x 10 7 cells, about 0.1 -
- 8.1 x 10 7 cells about 0.5- 8.1 x 10 7 cells, about 0.6- 8.1 x 10 7 cells, about 0.7- 8.1 x 10 7 cells, about 0.8-
- 9.1 x 10 7 cells about 0.4- 9.1 x 10 7 cells, about 0.5- 9.1 x 10 7 cells, about 0.6- 9.1 x 10 7 cells, about 0.7-
- 9.1 x 10 7 cells about 0.8- 9.1 x 10 7 cells, or about 0.9- 9.1 x 10 7 cells.
- a cell secreting a vaccine protein (e.g., gp96-lg) can be administered to a subject at about 0.1-1.1 x 10 7 cells, about 0.2- 1 .1 x 10 7 cells, about 0.3- 1 .1 x 10 7 cells, about 0.4- 1 .1 x 10 7 cells, about 0.5- 1 .1 x 10 7 cells, about 0.6- 1 .1 x 10 7 cells, about 0.7- 1.1 x 10 7 cells, about 0.8- 1 .1 x 10 7 cells, about or 0.9- 1 .1 x 10 7 cells, about 0.1-2.1 x 10 7 cells, about 0.2- 2.1 x 10 7 cells, about 0.3- 2.1 x 10 7 cells, about 0.4- 2.1 x 10 7 cells, about 0.5- 2.1 x 10 7 cells, about 0.6- 2.1 x 10 7 cells, about 0.7-
- 6.1 x 10 7 cells about 0.4- 6.1 x 10 7 cells, about 0.5- 6.1 x 10 7 cells, about 0.6- 6.1 x 10 7 cells, about 0.7-
- 6.1 x 10 7 cells about 0.8- 6.1 x 10 7 cells, about 0.9- 6.1 x 10 7 cells, about 0.1 -7.1 x 10 7 cells, about 0.2-
- 9.1 x 10 7 cells about 0.5- 9.1 x 10 7 cells, about 0.6- 9.1 x 10 7 cells, about 0.7- 9.1 x 10 7 cells, about 0.8-
- the cell based therapy can be administered to a subject one or more times (e.g once, twice, two to four times, three to five times, five to eight times, six to ten times, eight to 12 times, or more than 12 times).
- the cell based therapy as provided herein can be administered one or more times per day, one or more times per week, every other week, one or more times per month, once every two to three months, once every three to six months, or once every six to 12 months.
- the cell based therapy can be administered over any suitable period of time, such as a period from about 1 day to about 12 months.
- the period of administration can be from about 1 day to 90 days; from about 1 day to 60 days; from about 1 day to 30 days; from about 1 day to 20 days; from about 1 day to 10 days; from about 1 day to 7 days.
- the period of administration can be from about 1 week to 50 weeks; from about 1 week to 50 weeks; from about 1 week to 40 weeks; from about 1 week to 30 weeks; from about 1 week to 24 weeks; from about 1 week to 20 weeks; from about 1 week to 16 weeks; from about 1 week to 12 weeks; from about 1 week to 8 weeks; from about 1 week to 4 weeks; from about 1 week to 3 weeks; from about 1 week to 2 weeks; from about 2 weeks to 3 weeks; from about 2 weeks to 4 weeks; from about 2 weeks to 6 weeks; from about 2 weeks to 8 weeks; from about 3 weeks to 8 weeks; from about 3 weeks to 12 weeks; or from about 4 weeks to 20 weeks or any weekly increment of time in between.
- boosting doses of the cell based therapy as provided herein can be administered.
- a boosting dose can be administered about 10 to 30 days, about 15 to 35 days, about 20 to 40 days, about 25 to 45 days, or about 30 to 50 days after a priming dose.
- a secreatable vaccine protein ⁇ e.g., gp96-lg) and the T cell costimulatory fusion protein ⁇ e.g., a cell secreting OX40L-lg are administered 1 minute apart, 10 minutes apart, 30 minutes apart, less than 1 hour apart, 1 hour apart, 1 hour to 2 hours apart, 2 hours to 3 hours apart, 3 hours to 4 hours apart, 4 hours to 5 hours apart, 5 hours to 6 hours apart, 6 hours to 7 hours apart, 7 hours to 8 hours apart, 8 hours to 9 hours apart, 9 hours to 10 hours apart, 10 hours to 1 1 hours apart, 11 hours to 12 hours apart, 1 day apart, 2 days apart, 3 days apart, 4 days apart, 5 days apart, 6 days apart, 1 week apart, 2 weeks apart, 3 weeks apart, or 4 weeks apart.
- a regimen in which a first treatment of a cell secreting a T cell costimulatory fusion protein (e.g., OX40L-lg) is administered and subsequently a second treatment of a cell secreting a T cell costimulatory fusion protein (e.g., OX40L-lg) is administered and a cell secreting a vaccine protein [e.g., gp96-lg) is administered.
- a first treatment and the second treatment are about 3 days, or about 5 days, or about 1 week, or about 2 weeks or about 3 weeks apart.
- the first and second treatments are about two weeks apart.
- a single fixed dose of a cell secreting a T cell costimulatory fusion protein is administered and following the first dose of the cell secreting the T cell costimulatory fusion protein [e.g., OX40L-lg), a second dose is administered along with a fixed dose of a cell secreting a vaccine protein [e.g., gp96-lg) is administered.
- a single fixed dose of a cell secreting the secreatable vaccine protein e.g., gp96-lg
- an ascending dose of the cell secreting the T cell costimulatory fusion protein e.g., OX40L-lg
- the methods provided herein can be used for controlling solid tumor growth (e.g., breast, prostate, melanoma, renal, colon, or cervical tumor growth) and/or metastasis.
- the methods can include administering an effective amount of a cell based therapy as described herein to a subject in need thereof.
- the subject is a mammal (e.g., a human).
- the cell based therapies and methods provided herein can be useful for stimulating an immune response against a tumor. Such immune response is useful in treating or alleviating a sign or symptom associated with the tumor.
- treating is meant reducing, preventing, and/or reversing the symptoms in the individual to which a cell based therapy as described herein has been administered, as compared to the symptoms of an individual not being treated.
- methods described herein are to be used in concomitance with continuous clinical evaluations by a skilled practitioner (physician or veterinarian) to determine subsequent therapy. Such evaluations will aid and inform in evaluating whether to increase, reduce, or continue a particular treatment dose, mode of administration, etc.
- the methods provided herein can thus be used to treat a tumor, including, for example, a cancer.
- the methods can be used, for example, to inhibit the growth of a tumor by preventing further tumor growth, by slowing tumor growth, or by causing tumor regression.
- the methods can be used, for example, to treat a cancer such as a lung cancer.
- a cancer such as a lung cancer.
- the subject to which a compound is administered need not suffer from a specific traumatic state.
- the cell based therapy described herein may be administered prophylactically, prior to development of symptoms ⁇ e.g., a patient in remission from cancer).
- the terms "therapeutic” and “therapeutically,” and permutations of these terms, are used to encompass therapeutic, palliative, and prophylactic uses.
- treating or alleviating the symptoms is meant reducing, preventing, and/or reversing the symptoms of the individual to which a therapeutically effective amount of a composition has been administered, as compared to the symptoms of an individual receiving no such administration.
- the terms "effective amount” and "therapeutically effective amount” refer to an amount sufficient to provide the desired therapeutic ⁇ e.g., anti-cancer, anti-tumor, or anti-infection) effect in a subject (e.g., a human diagnosed as having cancer or an infection).
- Anti-tumor and anti-cancer effects include, without limitation, modulation of tumor growth (e.g., tumor growth delay), tumor size, or metastasis, the reduction of toxicity and side effects associated with a particular anti-cancer agent, the amelioration or minimization of the clinical impairment or symptoms of cancer, extending the survival of the subject beyond that which would otherwise be expected in the absence of such treatment, and the prevention of tumor growth in an animal lacking tumor formation prior to administration, i.e., prophylactic administration.
- administration of an effective amount of the cell based therapy can increase the activation or proliferation of tumor antigen specific T cells in a subject.
- the activation or proliferation of tumor antigen specific T cells in the subject can be is increased by at least 10 percent (e.g., at least 25 percent, at least 50 percent, or at least 75 percent) as compared to the level of activation or proliferation of tumor antigen specific T cells in the subject prior to the administration.
- Anti-infection effects include, for example, a reduction in the number of infective agents (e.g., viruses or bacteria).
- infective agents e.g., viruses or bacteria.
- administration of a cell based therapy as provided herein can stimulate the activation or proliferation of pathogenic antigen specific T cells in the subject.
- administration of the cell based therapy can lead to activation of antigen-specific T cells in the subject to a level great than that achieved by gp96-lg vaccination alone.
- an effective amount of a cell based therapy may be lowered or increased by fine tuning and/or by administering more than one dose ⁇ e.g., by concomitant administration of two different genetically modified tumor cells, or by administering the cell based therapy with another agent ⁇ e.g., an antagonist of PD-1 ) to enhance the therapeutic effect (e.g., synergistically).
- This disclosure therefore provides a method for tailoring the administration/treatment to the particular exigencies specific to a given mammal.
- Therapeutically effective amounts can be determined by, for example, starting at relatively low amounts and using step-wise increments with concurrent evaluation of beneficial effects.
- the methods provided herein thus can be used alone or in combination with other well-known tumor therapies, to treat a patient having a tumor.
- One skilled in the art will readily understand advantageous uses of the cell based therapies and methods provided herein, for example, in prolonging the life expectancy of a cancer patient and/or improving the quality of life of a cancer patient (e.g., a lung cancer patient).
- the invention provides for methods that further comprise administering an additional agent to a subject.
- the invention pertains to co-administration and/or co-formulation.
- administration of a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein, to a patient undergoing a treatment with a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein act synergistically when co-administered with another agent.
- administering act synergistically when co-administered with another agent and is administered at doses that are lower than the doses commonly employed when such agents are used as monotherapy.
- chemotherapeutic agents include, but are not limited to, alkylating agents such as thiotepa and CYTOXAN cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethiylenethiophosphoramide and trimethylolomelamine; acetogenins ⁇ e.g., bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; cally statin; CC-1065 (including its ad
- dynemicin including dynemicin A; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, ADRIAMYCIN doxorubicin (including morpholino- doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxy doxorubicin), epirubicin, 6-diazo-5-oxo-L-norleucine
- irinotecan Camptosar, CPT-11 (including the treatment regimen of irinotecan with 5-FU and leucovorin); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine; combretastatin; leucovorin (LV); oxaliplatin, including the oxaliplatin treatment regimen (FOLFOX); lapatinib (TYKERB); inhibitors of PKC-a, Raf, H-Ras, EGFR ⁇ e.g., erlotinib (Tarceva)) and VEGF-A that reduce cell proliferation and pharmaceutically acceptable salts, acids or derivatives of any of the above.
- the methods of treatment can further include the use
- the present invention pertains to anti-infectives as additional agents.
- the anti-infective is an anti-viral agent including, but not limited to, Abacavir, Acyclovir, Adefovir, Amprenavir, Atazanavir, Cidofovir, Darunavir, Delavirdine, Didanosine, Docosanol, Efavirenz, Elvitegravir, Emtricitabine, Enfuvirtide, Etravirine, Famciclovir, and Foscarnet.
- the anti-infective is an anti bacterial agent including, but not limited to, cephalosporin antibiotics (cephalexin, cefuroxime, cefadroxil, cefazolin, cephalothin, cefaclor, cefamandole, cefoxitin, cefprozil, and ceftobiprole); fluoroquinolone antibiotics (cipro, Levaquin, floxin, tequin, avelox, and norflox); tetracycline antibiotics (tetracycline, minocycline, oxytetracycline, and doxycycline); penicillin antibiotics (amoxicillin, ampicillin, penicillin V, dicloxacillin, carbenicillin, vancomycin, and methicillin); monobactam antibiotics (aztreonam); and carbapenem antibiotics (ertapenem, doripenem, imipenem/cilastatin, and meropenem).
- cephalosporin antibiotics ce
- the anti-infectives include anti-malarial agents (e.g., chloroquine, quinine, mefloquine, primaquine, doxycycline, artemether/lumefantrine, atovaquone/proguanil and sulfadoxine/pyrimethamine), metronidazole, tinidazole, ivermectin, pyrantel pamoate, and albendazole.
- anti-malarial agents e.g., chloroquine, quinine, mefloquine, primaquine, doxycycline, artemether/lumefantrine, atovaquone/proguanil and sulfadoxine/pyrimethamine
- metronidazole e.g., chloroquine, quinine, mefloquine, primaquine, doxycycline, artemether/lumefantrine, atovaquone/proguanil and sulfa
- the method comprises administration in combination with an agent that inhibits immunosuppressive molecules produced by tumor cells.
- the agent is an antibody against PD-1.
- the antibody against PD-1 is selected from nivolumab, pembrolizumab, pidilizumab, cemiplimab, AGEN2034, AMP-224, AMP-514, and PDR001 .
- the agent is an antibody against PD-L1.
- the antibody against PD-L1 is selected from Atezolizumab (Tecentriq), Avelumab (Bavencio), Durvalumab (Imfinzi), BMS-936559, and CK-301 .
- the agent is an antibody against CTLA-4.
- the antibody against CTLA-4 is selected from ipilimumab, tremelimumab, AGEN1884, and RG2077.
- the agent is an antibody against 0X40.
- the antibody against 0X40 is selected from PF-04518600, BMS-986178, INCAGN01949, MEDI0562, GSK1795091 , and GSK3174998
- the subject and/or animal is a mammal, e.g., a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, rabbit, sheep, or non-human primate, such as a monkey, chimpanzee, or baboon.
- the subject and/or animal is a non-mammal, such, for example, a zebrafish.
- the subject and/or animal may comprise fluorescently- tagged cells (with e.g., GFP).
- the subject and/or animal is a transgenic animal comprising a fluorescent cell.
- the subject and/or animal is a human.
- the human is a pediatric human.
- the human is an adult human.
- the human is a geriatric human.
- the human may be referred to as a patient.
- the human has an age in a range of from about 0 months to about 6 months old, from about 6 to about 12 months old, from about 6 to about 18 months old, from about 18 to about 36 months old, from about 1 to about 5 years old, from about 5 to about 10 years old, from about 10 to about 15 years old, from about 15 to about 20 years old, from about 20 to about 25 years old, from about 25 to about 30 years old, from about 30 to about 35 years old, from about 35 to about 40 years old, from about 40 to about 45 years old, from about 45 to about 50 years old, from about 50 to about 55 years old, from about 55 to about 60 years old, from about 60 to about 65 years old, from about 65 to about 70 years old, from about 70 to about 75 years old, from about 75 to about 80 years old, from about 80 to about 85 years old, from about 85 to about 90 years old, from about 90 to about 95 years old or from about 95 to about 100 years old.
- the subject is a non-human animal, and therefore the invention pertains to veterinary use.
- the non-human animal is a household pet.
- the non-human animal is a livestock animal.
- the subject is a human cancer patient that cannot receive chemotherapy, e.g., the patient is unresponsive to chemotherapy or too ill to have a suitable therapeutic window for chemotherapy ⁇ e.g., experiencing too many dose- or regimen-limiting side effects).
- the subject is a human cancer patient having advanced and/or metastatic disease.
- an "allogeneic cell” refers to a cell that is not derived from the individual to which the cell is to be administered, that is, has a different genetic constitution than the individual.
- An allogeneic cell is generally obtained from the same species as the individual to which the cell is to be administered.
- the allogeneic cell can be a human cell, as disclosed herein, for administering to a human patient such as a cancer patient.
- an “allogeneic tumor cell” refers to a tumor cell that is not derived from the individual to which the allogeneic cell is to be administered.
- the allogeneic tumor cell expresses one or more tumor antigens that can stimulate an immune response against a tumor in an individual to which the cell is to be administered.
- an "allogeneic cancer cell,” for example, a lung cancer cell refers to a cancer cell that is not derived from the individual to which the allogeneic cell is to be administered.
- a "genetically modified cell” refers to a cell that has been genetically modified to express an exogenous nucleic acid, for example, by transfection or transduction.
- the term “comprising” means that the process includes at least the recited steps, but may include additional steps.
- the term “comprising” means that the compound or composition includes at least the recited features or components, but may also include additional features or components.
- HS-130 which is a genetically engineered human lung adenocarcinoma cell line secreting OX40L-lg fusion protein, also (referred to as mHS-130 (B16F10) and HS-1 10 (referred to as mHS-1 10 (B16F10), generated from a B16F10-ova cell line, to determine a dose of mHS-130 when co-administered with a fixed dose of mHS-110.
- the starting dose of HS-130 in humans is based on the dose of mHS-130 that generates a minimum response (MABEL) in the mouse model based on the proportional use of mHS-110 in mice vs. the current dose of HS-110 in humans.
- MABEL minimum response
- mice In previous studies, the dose of murine HS-110 (mHS-1 10) in mice was determined to be 1 x 10 6 cells. It was further determined by quantitative ELISA that this number of cells secrete 290 ng of gp96-lg fusion protein per 24 hours. In order to determine the ratio of gp96-lg to OX40L-lg in this system, a murine form of HS-130 [mHS-130 (B16F10)] secreting the species specific OX40L-lg protein is generated. It has also been determined by quantitative ELISA that mHS-130 (B16F10) secretes 850 ng of OX40L-lg per 10 6 cells per 24 hours.
- Wild-type C57BL/6 mice are adoptively transferred with 10 6 OT-I* and 10 6 OT-ll FoxP3 - rf p congenic cells. These cells are transgenic CD8+ and CD4+ T cells designed to recognize a specific ovalbumin peptide. When transferred into wild-type mice these cells join the other murine lymphocytes in circulation until they encounter their cognate antigen and can be activated. Both of the murine vaccine cell lines mHS-1 10 (B16F10) and mHS-130 (B16F10) are engineered to produce ovalbumin, a protein not naturally expressed in the mouse.
- mice Two days after adoptive transfer the mice are vaccinated with mHS-1 10 (B16F10) and mHS-130 (B16F10) according to the doses described in the study protocol below (Table 1). Mice are followed for 14 days. Periodically, peripheral blood is collected from the tail vein and analyzed for the frequency of CD8+ and CD4+ T cells by flow cytometry. The development of CD4+ FoxP3+ T regulatory subset of cells are further characterized in this system by the induction of red fluorescent protein upon expression of the master regulator FoxP3. This helps with studying the influence of vaccination platform on antigen specific T lymphocytes over time, that have been shown to be critical for effective tumor reduction and control. Additionally, the weight of each animal is tracked at the time of blood draw, as an indicator of overall health throughout the study.
- the luciferase activation was measured after 5 hours using Bio-Glo reagent from Promega and the relative luminescence was measured using a luminometer.
- the ECso values were calculated using a four-parameter logistic curve analysis. The calculated ECso values indicate that the activity of the mouse and human derived OX40L are very similar against the human 0X40 receptor.
- Example 2 Phase 1 Clinical Trial, Dosage Form, Route of Administration and Dosing Regimen
- the drug product is a viable whole cell vaccine that has been irradiated to render cell replication-incompetent while expressing the co-stimulatory fusion protein OX40L-lg, which is the ligand for the 0X40 receptor, a member of the TNF-receptor superfamily.
- the drug product is a viable whole cell vaccine derived from a human lung adenocarcinoma cell line.
- the cell line is transfected with a 7192 bp plasmid cDNA‘pcDNA3.4 OX40L-lg' stably expressing the cDNA for OX40L-lg to develop an irradiated whole cell vaccine as shown in FIG. 1.
- HS-110 refers to (viagenpumatucel-L); genetically engineered human lung adenocarcinoma cell line secreting gp96-lg fusion protein, and HS-130 refers to genetically engineered human lung adenocarcinoma cell line AD100, secreting OX40L-lg fusion protein.
- HS-130 is genetically modified, viable, replication-incompetent cancer cells, designed to stimulate an immune response when administered into the intradermal layers of the skin.
- the purpose of the study is to study the safety and immunological response associated with the treatment.
- patients who meet the inclusion/exclusion criteria receive escalating doses of the HS-130 cells using a 3 + 3 design.
- the first cohort start treatment with a single fixed dose of HS-130 based on the minimum anticipated biological effect level (MABEL) established in animal models.
- MABEL minimum anticipated biological effect level
- the patient After a safety assessment interval of 2 weeks following the first dose of HS-130, the patient is administered the same dose of HS-130 along with a fixed dose of HS-110 cells (1 x 10 7 cells, which is an established safe dose in the ongoing Phase 2 study of HS-110). In the absence of any safety issues 2 weeks after the combination treatment, the patient can continue the combination treatment of HS-110 + HS-130 administered bi-weekly for 6 months or until disease progression, death, patient withdrawal of consent, investigator decision to remove patient, or intolerable toxicity, whichever occurs first.
- the dose escalation committee recommends enrollment at the next higher dose level.
- the conduct and completion of each subsequent dose cohort follows in similar fashion until DLT occurs in 2 patients among a total of up to 6 treated patients at a dose level.
- the sponsor may decide to discontinue further dose escalation. Any patient who does not complete the first cycle of treatment (i.e. at least one combination dose) is replaced. It is predicted that 4 dose levels of HS-130 are explored, and 12 to 24 patients are enrolled on the trial. All patients are monitored for extensive safety assessment including serum cytokines/chemokines, and immune phenotype profiling of immune cell subsets by flow cytometry. The immune response is evaluated by ELISPOT using HS-110 lysate and HS-110 specific peptides for IFNy and Granzyme B production.
- AEs adverse events
- CTCAE version 5 is used to grade all toxicities.
- Inclusion Criteria Patients must meet all of the following inclusion criteria before they are allowed to participate in the trial: Patients with select solid tumor types (defined as those having CTA overexpression overlap with at least 10 CTAs overexpressed by HS-110) who have failed available standard therapy or who are not candidates for standard therapy, and for whom, in the opinion of the investigator, experimental therapy with HS-110 + HS-130 may be beneficial. Age 3 18 years. Have an acceptable organ function. Have an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1. Life expectancy of at least three months. Have fresh or archival tumor tissue available at screening. Patients, both females and males, of childbearing/reproductive potential must agree to use adequate contraception while included in the trial and for six months after the last treatment with HS-130 and/or HS-110. Patients must sign the informed consent form.
- EOG Eastern Cooperative Oncology Group
- Exclusion Criteria If any of the following apply, the patient must not enter the trial: Clinically significant cardiac disease, congestive heart failure and/or uncontrolled hypertension. Known or clinically suspected leptomeningeal disease. Stable, previously treated metastases in the brain or spinal cord, are allowed as long as these are considered stable (by CT or MR), and not requiring systemic corticosteroids. History of 3 grade 3 allergic reactions as well as known or suspected allergy or intolerance to any agent given in the course of this trial, live cell therapies, or live vaccines. History of suspected cytokine release syndrome (CRS). Known immunodeficiency disorders. Ongoing or current autoimmune disease (history of checkpoint inhibitor immune related adverse events are permissible if resolved).
- CRS cytokine release syndrome
- Prior therapy must be stopped within four weeks before first infusion in the study, or 5 half-lives, or twice the duration of the biological effect of the investigational product (whichever is shortest).
- Adjuvant anti-hormonal treatment(s) for prior breast cancer or prostate cancer are allowed.
- Prior curable cancer with complete remission for >2 years is allowed. Any other ongoing significant, uncontrolled medical condition. Received a live vaccine within 30 days prior to first dose of study drug.
- Clinically significant active viral, bacterial or fungal infection requiring: Intravenous treatment with antimicrobial therapy completed less than two weeks prior to first dose, or Oral treatment with antimicrobial therapy completed less than one week prior to first dose.
- Prophylactic treatment e.g., for dental extractions
- Known positive serology for human immunodeficiency virus (HIV), hepatitis B, or hepatitis C (except in cases of immunity after cured infection).
- Substance abuse, medical, psychological or social conditions that may interfere with the patient's participation in the trial or evaluation of the trial result. Women who are pregnant or breast feeding.
- Dose and Mode of Administration Each patient is administered an intradermal injection of HS-130 on Day 1 and another intradermal injection of HS-130 at the same dose with a fixed dose of HS-110 (1 x 10 7 cells) on Day 15. Patients not progressing may continue treatment at the discretion of the investigator.
- DLT Dose Limiting Toxicity: DLT is defined as any non-acceptable (as defined below) treatment related toxicity (i.e., not attributable to the active disease, disease-related processes under investigation or intercurrent illness) observed during the first 28 days of study treatment. Ongoing safety events beyond Cycle 1 is reviewed across all cohorts during the study to help inform dose escalation decisions.
- DLT includes: Hematological toxicities 3 CTCAEv5 grade 3.
- DLT excludes: Grade 3 self-limited or medically controllable toxicities ⁇ e.g., fever without 3 Grade 3 neutropenia, nausea, vomiting, diarrhea, fatigue). Electrolyte disturbances that are managed to grade 1 or less with supplemental therapy.
- DMC Data Monitoring Committee
- Duration of Treatment Upon completion of the first treatment cycle (i.e. 4 weeks, with one dose of HS-130 on Day 1 , and one combined dose with HS-110 on Day 15), in the absence of disease progression or unacceptable toxicity, patients may continue to be treated with the combination of HS-130 + HS-110 at the same dose bi-weekly for 6-months or until disease progression, death, patient withdrawal of consent, investigator decision to remove patient, or intolerable toxicity, whichever occurs first. [00210] Criteria for Evaluation:
- Safety and Tolerability Measured by the frequency of treatment emergent adverse events (TEAEs)/serious adverse events (SAEs), including clinically significant (CS) abnormal laboratory parameters, ECGs, PEs, and vital signs in patients receiving at least 1 dose of study drug.
- TEAEs treatment emergent adverse events
- SAEs serious adverse events
- CS clinically significant abnormal laboratory parameters
- ECGs ECGs
- PEs vital signs in patients receiving at least 1 dose of study drug.
- MTD Maximum Tolerated Dose
- Patient disease status is monitored by clinical and radiological assessment as per the institutional standard. Patient clinical response is evaluated as complete response, partial response, stable disease, or progressive disease per Investigator assessment.
- Safety All patients who have received any component of study treatment are considered evaluable for safety. Safety is assessed by means of physical examination, weight, vital signs, performance status, laboratory evaluations (hematology, biochemistry including cytokines and C-reactive protein), electrocardiogram (ECG), and recording of concurrent illness/therapy and adverse events. CTCAE version 5 are used to grade all toxicities. All related adverse events are monitored until resolution. Patients are monitored for safety and concomitant medications throughout the study.
- Cytokines ⁇ e.g., IFNy, IL-1 b, IL-2, IL-4, IL-6, IL-10, IL-12 p70, IL-17A, TNFa) are be monitored once after each dose and repeatedly in relation to clinically observed reactions after drug injections.
- Example 3 Mouse HS-110 (B16F10-OVA-gp96) and mouse HS-130 (B16F10-OVA- OX40L) immunization at a“narrow” dose-range to correlate CD8+ T-cell expansion to tumor growth
- mice HS-110 mHS-110; B16F10-OVA-gp96-lg
- mouse HS-130 mHS-130; B16F10-OVA-OX40L
- anti-tumor T-cell expansion correlates to tumor growth control, in vivo.
- HS-110 is a lung adenocarcinoma cancer cell line.
- HS- 1 10 secretes gp96-lg, which presents antigens to prime and expand CD8+ T cell responses.
- Preclinical studies have suggested that the addition of secreted T cell costimulatory molecules, OX40L-lg in combination with Gp96-lg in a cellular system expressing a defined antigen, enhances T cell immunity and results in elimination of tumors (Fromm G et al., Cancer Immunol Res. 2016 Sep 2;4(9):766-78.).
- mice HS- 130 a mouse cell line was devised that secretes only OX40L-lg, herein referred to as "mouse HS- 130” or “mHS-130”, which was used in combination with mHS-110 (See Example 1, above).
- mHS-130 a mouse cell line was devised that secretes only OX40L-lg, herein referred to as "mouse HS- 130” or “mHS-130”, which was used in combination with mHS-110 (See Example 1, above).
- the best ratio of mHS-110 to mHS-130 for the generation of a primary and secondary CD8+ or CD4+ T cell pool was investigated in the following experiments.
- the experiments of this example further included a tumor challenge arm to better understand how in vivo expansion of CD8+ T cells correlates to anti-tumor immune responses and tumor growth control.
- T-cell receptor (TCR) transgenic mouse CD8+ (OT-I) cells were isolated from OT-I-GFP bred mice using Easy Sep Mouse CD8 T Cell isolation kit (cat # 19853A) and injected into each C57BL/6 mouse intravenously (i.v) through lateral tail vein with 1 million OT-I cells suspended in HBSS (GIBCO 14175-095).
- mice Two days after injecting OT-I, all the mice were tail bled for baseline and 4 hours later, mHS-1 10 (B16F10-OVA-gp96-lg) cells and mHS-130 (B16F10-OVA-OX40L-lg) were treated with 10 pg/mL of Mitomycin-C (Sigma-Aldrich cat# M0503) for 3 hours and given intraperitoneally (i.p.) to each group accordingly.
- Mice were divided into 7 groups with 5 mice per group, except for the parental group which had 3 mice. Animals were dosed based on nanogram expression level (per 10 6 cells per 24hr) of gp96-lg or OX40L-lg per the design shown in FIG. 3 and Table 2 below.
- mice were tail bled consecutively on days 3, 5, 7, 10, and 14 post-immunization, and days 17, 19, 21 , 24, 28, 33, 38 and 41 post-boosts into heparinized PBS (10 units/ml), and lysed using ACK lysis buffer (150 mM NhUCI, 100 mM KHCO3 and 10 mM EDTA 0.2 Na, pH 7.2) for 3 minutes, and neutralized with 1 X PBS.
- ACK lysis buffer 150 mM NhUCI, 100 mM KHCO3 and 10 mM EDTA 0.2 Na, pH 7.2
- Cells were washed, resuspended in 50 pi of BD Cytofix/Cytoperm, and incubated at 4°C for 20 min before another two washes and staining with anti-IFN-g. Cells were washed once before acquisition and analysis of fluorescence. Analysis was done using FlowJo software (Tree Star Inc.); events were gated for live lymphocytes on FSC x SSC followed by CD8+ cells using CD8 x CD3 and displayed as CD8 x IFN-y.
- B16F10-OVA tumor challenge and volume calculations Melanoma B16F10 cells were harvested and resuspended at a concentration of 5x10 5 cells/100 mI in a volume of 80 mI HBSS and 20 mI Matrigel. C57BL/6 mice were subcutaneously injected with 100 mI of B16F10 cells (5x10 5 cells/mouse) on the inner abdomen 29 days post-OT-1 transfer and 28 days post-primary vaccination, designated "day 28”, as shown in study design (FIG. 3). The tumor size was measured and documented every 3 days with a caliper, starting on day 7, and calculated using the formula (AxB; A as the largest and B as the smallest diameter of tumor). Tumor growth was documented as standard error mean. To record the survival of the tumor-bearing mice, either natural death or a tumor volume greater than 450 mm 2 leading to death was counted as death. Each experimental group included five animals.
- TILs Tumor Infiltrating Lymphocytes
- ELISPOT Assay Splenocytes were harvested and red blood cell lysis buffer (cat#36858500, Roche) was used to eliminate red blood cells. Cells were washed in IMDM medium and pelleted. Counted cells were resuspended in IMDM with 10% FBS. Each ELISPOT well received 1 million cells, in a total volume of 200 pi.
- Treatments included the use of B16F10-OVA parental line lysates (vial of 10 million cells; freeze-thawed three times) at a 10-fold dilution, immunodominant epitopes for B16F10 tumors: "gp100/pmel” (EGSRNQDWL (SEQ ID NO:38)), "TRP-1/gp75” (TWHRYHLL (SEQ ID NO:39)), TRP-2 (SVYDFFVWL (SEQ ID NO:40)) and the MHC-I restricted peptide for H2 b haplotype for OVA (SIINFEKL (SEQ ID NO:41)). All peptides were used at a final concentration of 10 ug/mL.
- PMA/lonomycin was used as a positive control at "1 to 400 dilution” 100 nM of PMA, 1.6 mM ionomycin (500x stock from eBioscience, cat#00-4970-03, and whole chicken ovalbumin protein "OVA” at 250 pg/ml.
- Interferon-gamma capture ELISPOT assay was carried out as previously published (Klinman DM et at., Current Protocols in Immunology. 1994 June (1): 6.19.1-6.19.8). Briefly, MAIP N45 Milipore 96-well filtration plates were coated with anti-mouse IFN-g purified monoclonal antibody (clone AN18) in PBS saline at 10 pg/mL, 50 mI/well, at 4°C, overnight. The next day, cells were washed in IMEM medium plus 10% FBS at 200 ul/well, washed three times, then blocked for 1 hour at 25°C.
- Na ' ive OT-I GFP+ CD8+ T cells expressing ⁇ /a2L/b5 and specific for H-2K b /OVA25/-264 were adoptively transferred into na ' ive WT C57BL/6 mice.
- Recipient mice were injected with mHS-110 at a fixed dose of 1 million cells (290 ng of gp96-lg) with different ratios of mHS-130, in which a 1 to 1 ratio of mHS-1 10 to mHS-130 was equivalent to 290 ng of gp96-lg to 290 ng of OX40L.
- OT-I GFP+ CD8+ T cells were analyzed in the blood on days 0, 3, 5, 7, 10, and 14, representing the acute expansion phase and the contraction phase of CD8+ T cell responses. Based on accumulation of the transferred cells, the generation of the primary effector pool was found to peak on day 7 (FIG. 4, FIG. 6A, FIG. 6B) and contract by day 14 post-vaccination (FIG. 6A, FIG. 6B). Significant increases were found on day 7 in the 1 to 1 , the 1 to 3 and the 1 to 10 ratios of mHS-110 to mHS-130 (FIG. 6A, FIG. 6B) (*p ⁇ 0.05; **p ⁇ 0.01). Furthermore, the 1 to 1 ratio had significant increases on days 10 and 14 post-vaccination (*p ⁇ 0.05).
- TILs Tumor infiltrating lymphocytes
- the proportion of CD8+ TILs in the 1 to 1 and 1 to 10 groups were 51-fold and 118-fold, respectively, over that of the 1 to 0.1 vaccination ratio of mHS-110 to mFIS- 130 (FIG. 8A, FIG. 8B).
- the 1 to 1 group had two animals that failed to establish stable tumors (full tumor growth inhibition), and the 1 to 10 group had three animals with full tumor growth inhibition. For this reason, proper statistics for TIL percentages cannot be performed with less than three samples, thus the 1 to 10 group is ineligible for statistical analysis.
- Tumor volume was measured over time, from the point of tumor cell inoculation until end of study, a total of 25 days of logarithmic growth. Since this was a protection, prophylaxis, study, tumor delay was measured, rather a therapeutic response to established tumors.
- FIG. 10A only two ratios, 1 to 1 (290 ng of gp96-lg to 290 ng of OX40L-lg) and 1 to 10 (290 ng of gp96-lg to 2900 ng of OX40L-lg) of mHS-110 to mHS-130, gave a consistent and significant tumor growth inhibition as compared to the mHS-110 group alone, with greatest separation observed for days 21 -25 (**p ⁇ 0.01).
- FIG. 10B Individual tumor growth curves are shown in FIG. 10B to show individual animal variance. Measuring end-of-study tumor mass confirmed what tumor volume had demonstrated, in that both the 1 to 1 and 1 to 10 ratio gave the best response, as compared to mHS-110 alone (FIG. 9). For tumor volume, the 1 to 1 , and 1 to 10 ratio were not significantly different.
- Elevated expression of PD-1 was seen in those groups with the greatest tumor burden (1 to 0.1 , mHS-1 10 alone and parental B16F10- OVA alone), which suggested that cells have become exhausted from continued anti-tumor fighting, but those that have received proper stimulation with OX40L, via mHS-130 vaccination, show lower PD-1 expression and less tumor burden. This may explain why the 1 to 1 and 1 to 10 ratio dose groups showed the lowest PD-1 expression in the spleen. Expression of PD-1 was also measured in the tumor, by looking at TILs, but the frequency was below the limit of quantitation making analysis difficult (data not shown).
- Example 4 Study ofgp96-lg (mHS-110) to OX40L-lg (mHS-130) dose ratios
- HS-1 10 is a lung adenocarcinoma cancer cell line that secretes gp96-lg, which presents antigens to prime and expand CD8+ T cell responses.
- a mouse cell line (mHS-130) was developed that secretes only OX40L-lg.
- mHS-130 secreting gp96-lg fusion protein
- mHS-1 10 secreting OX40L-lg fusion protein
- An objective of the study was to determine ratio of mHS- 1 10 to mHS-130 most suitable for generation of a primary and secondary CD8+ or CD4+ T cell pool.
- this study tested in tumor-bearing animals a variable ratio, in a dose-escalation manner, to determine the ratio(s) and dose(s) that result in the most effective CD8+ T-cell expansion and tumor growth inhibition combination.
- FIG. 13 illustrates a design of the present study.
- T-cell receptor (TCR) transgenic mouse CD8+ (OT-I) cells were isolated from in-house bred OT-I-GFP mice using Easy Sep Mouse CD8+ T Cell isolation kit (cat # 19853A) and injected into each C57BL/6 mouse intravenously (i.v.) through lateral tail vein with 1 million OT-I cells suspended in HBSS (GIBCO 14175-095).
- mice Two days after injecting OT-I, all the mice were tail bled for baseline and 4 hours later, mHS-110 (B16F10-OVA-gp96-lg) cells and mHS-130 (B16F10-OVA-OX40L-lg) were treated with 10 pg/mL of Mitomycin-C (Sigma-Aldrich cat# M0503) for 3 hours and given intraperitoneally (i.p.) to each group accordingly. Mice were divided into 10 groups with 5 mice per group. Three different treatment ratios were provided with escalating doses within each ratio group and all compared against mHS-1 10 (gp96-lg) alone, the control group (ratio 1 to 0).
- Mitomycin-C Sigma-Aldrich cat# M0503
- mice were dosed based on nanogram expression level (measured as ng/10 6 cells/24 hrs) of gp96-lg or OX40L-lg, as shown in Tables 3 and 4 below. As shown in the study design FIG. 13, dose ratios of gp96-lg to OX40L-lg were 1 :1.3, 1 :2.5, 1 :5, and 1 :0 (mHS-110 (gp96-lg) alone), and each dose was tested at three different dose levels ("low,” “medium,” and "high”).
- mice were boosted on days 14 and 31 with the same ratios of mHS-110 and mHS-130 as in the primary phase, and OT-I GFP+ CD8+ T cells were analyzed in the blood days post-challenge. T umor was provided on day 28. Consecutive bleeds were collected from the peripheral blood and analysis was performed by flow cytometry on both exogenous, adoptively transferred, OT-1 and endogenous CD8+ and CD4+ T-cells for activation, and short (SLECs) and long-term (MPECs) memory markers as outlined in the methods section. Tumor growth kinetics, response rates and infiltrating lymphocytes, were also quantitated.
- SLECs short
- MPECs long-term
- the amount of murine gp96 protein expressed by the mHS-110 cells was determined by ELISA. For each sample to be tested, one million B16F10-Ova9 parental and mHS-110 cells were plated in a 6-well tissue culture plate in a total volume of 1 ml each. Cells were incubated at 37°C with 5% CO2 for 24 hours at which point the supernatants were harvested. Supernatants were then centrifuged at 2500 rpm for 5 minutes to pellet any cell debris. Clarified supernatants were then transferred to new 1.5 ml tubes and stored at -80°C. Each sample tested was from a fresh vial of mHS-1 10 cells thawed and expanded.
- 96-well plates (Corning, cat# 9018) were coated with 2 ug/ml of sheep anti-gp96 (R&D Systems, cat# AF7606) in carbonate-bicarbonate buffer. Plates were sealed and stored at 4°C overnight. Plates were then washed 4 times with 1X TBST (VWR, cat# K873) and then blocked with 1X casein solution (Sigma-Aldrich, cat# B6429) for 1 hour at room temperature.
- 1X TBST VWR, cat# K873
- 1X casein solution Sigma-Aldrich, cat# B6429
- a human gp96-mouse Fc standard (Thermo Fisher Scientific, lot# 2065447) was prepared in IMDM (Gibco, cat# 12440-053) with 10% FBS (Gibco, cat# 10082-147).
- IMDM Gibco, cat# 12440-053
- FBS Gibco, cat# 10082-147
- a 2000 ng/ml human-gp96-mFc standard solution was made and 2-fold serial dilutions were performed down to 1 .95 ng/ml.
- Sample supernatants were loaded onto the ELISA plates starting at a 1 :2 dilution and then 2-fold serial dilutions were performed to a highest dilution of 1 :16.
- the amount of mouse OX40L protein expressed by the mHS-130 cells was also determined by ELISA.
- 96-well plates were coated with 2.5 ug/ml His-tagged mouse 0X40 protein (Aero Biosystems, cat# OXO-M5228) in PBS. Plates were sealed and stored at 4°C overnight. Plates were then washed 4 times with 1X TBST and then blocked with 1 % BSA (Sigma-Aldrich, cat# A2153) for 1 hour at room temperature. The plates were then washed 4 times with 1X TBST and a mouse lgG1-mouse OX40L standard (Thermo Fisher Scientific, lot# 2214217) was prepared in IMDM with 10% FBS.
- Table 3 Expression of mouse gp96-lg and OX40L-lg from mHS-110 and mHS-130, respectively, per million cells in a 24-hour period
- Table 4 Expression level of active biologic protein per cell type and resulting ratio for each group
- mice were tail bled consecutively on days 3, 5, 7, 10, 12, and 14 post-immunization and days 17, 19, 21 , 24, 26, 28, 33, 38, 41 , 45, 48, and 54 into heparinized PBS (10 units/ml) and lysed using ACK lysis buffer (150 mM NhUCI, 100 mM KHCO3 and 10 mM EDTA 0.2 Na, pH 7.2) for 3 minutes and neutralized with 1 X PBS.
- ACK lysis buffer 150 mM NhUCI, 100 mM KHCO3 and 10 mM EDTA 0.2 Na, pH 7.2
- Consecutive bleeds were collected from the peripheral blood and analysis was performed by flow cytometry on both exogenous, adoptively transferred, OT-1 and endogenous CD8+ and CD4+ T-cells for activation, and short (SLECs) and long term (MPECs) memory markers as outlined in the methods section. Tumor growth kinetics, response rates and infiltrating lymphocytes, were also quantitated.
- Cells were washed, resuspended in 50 pi of BD Cytofix/Cytoperm, and incubated at 4°C for 20 min before another two washes and staining with anti-IFN-g. Cells were washed once before acquisition and analysis of fluorescence. Analysis was done using FlowJo software (Tree Star Inc.); events were gated for live lymphocytes on FSC x SSC followed by CD8+ cells using CD8 x CD3 and displayed as CD8 x IFN-y.
- B16F10-OVA tumor challenge and volume calculations Melanoma B16F10 cells were harvested and resuspended at a concentration of 5x10 5 cells/100 mI in a volume of 80 mI HBSS and 20 mI Matrigel. C57BL/6 mice were subcutaneously injected with 100 mI of B16F10 cells (5x10 5 cells/mouse) on the inner abdomen 29 days post-OT-1 transfer and 28 days post-primary vaccination, designated "day 28”, as shown in the study design (FIG. 13). The tumor size was measured and documented every 3 days with a caliper, starting on day 7, and calculated using the formula (AxB; A as the largest and B as the smallest diameter of tumor). Tumor growth was documented as standard error mean. To record the survival of the tumor-bearing mice, either natural death or a tumor volume greater than 450 mm 2 leading to death was counted as death. Each experimental group included five animals.
- TILs Tumor Infiltrating Lymphocytes
- ELISPOT Assay Splenocytes were harvested and red blood cell lysis buffer (cat#36858500, Roche) was used to eliminate red blood cells. Cells were washed in IMDM medium and pelleted. Counted cells were resuspended in IMDM with 10% FBS. Each ELISPOT well received 1 million cells, in a total volume of 200 mI.
- Treatments included the use of B16F10-OVA parental line lysates (vial of 10 million cells; freeze-thawed three times) at a 10-fold dilution, immunodominant epitopes for B16F10 tumors: "gp100/pmel” (EGSRNQDWL (SEQ ID NO:38)), "TRP-1/gp75” (TWHRYHLL (SEQ ID NO:39)), TRP-2 (SVYDFFVWL (SEQ ID NO:40)) and the MHC-I restricted peptide for H2b haplotype for OVA (SIINFEKL (SEQ ID NO:41)). All peptides were used at a final concentration of 10 ug/mL.
- PMA/lonomycin was used as a positive control at "1 to 400 dilution” 100 nM of PMA, 1.6 mM ionomycin (500x stock from eBioscience, cat#00-4970-03, and whole chicken ovalbumin protein "OVA” at 250 pg/ml.
- Interferon-gamma capture ELISPOT assay was carried out as previously published (Klinman et al., Current Protocols in Immunology. 1994 June (1 ): 6.19.1-6.19.8). Briefly, MAIP N45 Milipore 96- well filtration plates were coated with anti-mouse IFN-g purified monoclonal antibody (clone AN18) in PBS saline at 10 mo/hh ⁇ , 50 mI/well, at 4°C, overnight. The next day, cells were washed in IMEM medium plus 10% FBS at 200 ul/well, washed three times, then blocked for 1 hour at 25°C.
- the plate was then washed in PBS to remove the tween, then 100 mI of the AEC substrate (Vector kit) added in 100 mM of TRIS buffer at pH 8.2 was added to each well to allow for development of spots. At the end of the incubation, 10-20 minutes, all plates incubated for the same time, the reaction as stopped by rinsing the plates with tap water and air dried.
- Plates were scanned and counted in an AID Autoimmun Diagnostika GMBPI iSpot monochromatic ELISPOT reader (2016). Counting parameters for each plate remained consistent across all plates that were developed together as to prevent bias, as spot development, density and magnitude is influence by the operator, reagent lots and final substrate development time.
- FIG. 14 illustrates anti-tumor CD8+ OT-I T cell expansion in the peripheral blood with prime and boost Immunization of different ratios and dose combinations of mHS-110 and mPIS-130.
- Recipient mice were injected with mHS-110 and mHS-130 at different ratios and doses of gp96-lg to OX40L-lg.
- OT-I GFP+ CD8+ T cells were analyzed in the blood on days 0-54 days post-vaccination. Mice were boosted on day 14 with the same ratios of mHS-1 10 and mHS-130 as in the primary phase, and OT-I GFP+ CD8+ T cells were analyzed in the blood days post-challenge.
- the ratio 1 :1.3 ratio of mHS-1 10 to mHS-130 (high dose) results in the peak of CD8+ OT-l T cell expansion at day 7 and remains higher than other dose ratios through day 54.
- FIG. 15A illustrates gating strategy for flow cytometry experiments of the study of FIG. 13.
- Recipient mice were injected with a mHS-110 and mHS-130 at different ratios and doses of gp96-lg to OX40L-lg.
- OT-I GFP+ CD8+ T cells were analyzed in the blood on days 0-54 days post-vaccination. Mice were boosted on day 14 and 31 with the same ratios of mHS-110 and mHS-130 as in the primary phase, and OT-I GFP+ CD8+ T cells were analyzed in the blood days post-challenge. Tumor was provided on day 28.
- FIG. 15B illustrating expansion of OT-1 cells on days 7 and 17, shows that the dose ratio 1 to 1.3 provided the best response with the greatest expansion seen for the high dose, 339 ng gp96 to 441 ng OX40L.
- the dose ratio 1 to 1.3 provided the best response with the greatest expansion seen for the high dose, 339 ng gp96 to 441 ng OX40L.
- Tumor-specific, TRP2 tetramer positive CD8+ T-cells increased with treatment in a dose- dependent manner and showed a significant increase over treatment with 38 ng of gp96-lg via mHS-110 alone (FIG. 21)
- the transferred CD8+ OT-1 + eGFP cells at day 55, only the 1 to 1 .3 ratio of 339 ng gp96-lg to 441 ng OX40L-lg dose (high dose) showed any presence of expanded subsets on day 55 in both the spleen and blood, as shown in FIG. 22.
- FIG. 23 illustrates an increased percentage of exhausted CD4+ T-cells staining positive for PD-1 in the spleen (% of CD3+ CD4+ PD-1 + T cells) on day 55.
- the percentage of effector memory CD4+ T-cells in the spleen of treated and tumor-burden mice changed with vaccination, in an OX40L dose-dependent manner, as shown in FIG. 24.
- This increase in the percentage of activated CD4+ T- cells correlated with an increase proportion of CD4+ T-cell infiltrating the tumor (TILs), and was dose dependent (see FIG. 26).
- both CD4+ and CD8+ T-cell subsets are expanded with gp96/OX40L-lg co-vaccinations that correlate with increased TIL percentages and tumor growth inhibition; 2) the best dose combination for long-term survival and expansion of tumor specific CD8+ T- cells cells is the 1 to 1.3 ratio at a dose of 339 ng of gp96-lg to 441 ng of OX40L-lg; and 3) a dose determines tumor growth inhibition, with 38 ng of gp96-lg to 50 ng of OX40L-lg being the no-observed effect level (NOEL) and 1 13 ng gp96-lg to 147 ng of OX40L-lg being the minimum active biological effect level (MABEL) for this dose combination, in mice.
- NOEL no-observed effect level
- MABEL minimum active biological effect level
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Oncology (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Toxicology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present disclosure provides methods of treatment with cells having a vaccine (e.g., gp96-lg) and cells having a T-cell co-stimulatory molecule.
Description
COMBINATION CELL-BASED THERAPIES
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to and the benefit of U.S. Provisional Patent Application No. 62/739,814, filed on October 1 , 2018, and U.S. Provisional Patent Application No. 62/807,783, filed on February 20, 2019, the entire contents of which are herein incorporated by reference herein in their entireties.
FIELD OF THE DISCLOSURE
[0002] The disclosure is directed to methods of treatment with cells having a vaccine {e.g., gp96-lg) and cells having T-cell co-stimulatory molecules.
DESCRIPTION OF THE TEXT FILE SUBMITTED ELECTRONICALLY
[0003] The contents of the text file submitted electronically herewith are incorporated herein by reference in their entirety: A computer readable format copy of the Sequence Listing (filename: HTB- 030_ST25.txt; date recorded: September 30, 2019; file size: 79.2 KB).
BACKGROUND
[0004] Cancer is characterized by a progressive acquisition of genetic mutations that lead to intrinsic dysregulation of cell growth and death. Once a cell has acquired enough mutations, typically thought to be at least six, it no longer is responsive to intrinsic or extrinsic signals that would restrain its growth or trigger apoptosis. As tumors arise from host cells, the body's immune system is initially tolerant to those cells. A cell that acquires an immunogenic mutation, can be sought out and destroyed by the host immune system in a process known as immunosurveillance. Immune checkpoint therapy, which targets regulatory pathways in T cells to enhance anti-tumor immune responses, has led to important clinical advances and provides a new defense against cancer. Moreover, vaccines may contribute to this defense by also enhancing anti-tumor immune responses. Accordingly, it is possible that combination therapies including combinations or sub-combinations of one or more checkpoint inhibitors, one or more vaccines, and one or more T cell costimulatory molecules may expand the base of cancer patients that can benefit from immunotherapy.
SUMMARY
[0005] Immunotherapies aimed at including combinations of one or more vaccines, one or more T cell costimulatory molecules, and one or more checkpoint inhibitors may expand the base of cancer patients that can benefit from such therapies. Vaccines may contribute to this response by increasing both the frequency of tumor-antigen specific CD8+ T cells and also the number of tumor antigens recognized by those CD8+ T cells. T cell costimulatory molecules may enhance the response by further increasing the frequency and/or enhancing the activation of tumor antigen-specific T cells, and also by increasing the expression of tumor-killing effector molecules by CD8+T cells. When used in combination with checkpoint inhibitors, it may be possible to generate a broad range of highly activated CD8+ T cells that will be able to infiltrate tumors and will not be inhibited by various checkpoint pathways once infiltration has occurred. This present disclosure is based, at least in part, on the discovery that a combination of a vaccination, e.g., gp96-lg vaccination, and T cell costimulation with one or more agonists of 0X40, ICOS, 4-1 BB, TNFRSF25, CD40, CD27, and/or GITR, among others, provides a synergistic anti-tumor benefit. Pre- clinical models have evaluated independent compositions of gp96-lg vaccines combined with agonistic antibodies targeting 0X40, ICOS, 4-1 BB, and TNFRSF25, and demonstrated variable effects on mechanistic and anti-tumor complementarity. The methods described herein provide a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein, {e.g., gp96-lg) expression vector, wherein the patient is undergoing a treatment with a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein, including, without limitation, fusion proteins such as ICOSL-lg, 4-1 BBL-lg, TL1 A-lg, OX40L-lg, CD40L-lg, CD70-lg, or GITRL-lg to provide T cell costimulation.
[0006] In some embodiments, the methods described herein secrete fusion proteins that work synergistically. The effect of a locally secreted T-cell costimulatory fusion protein [i.e., OX40L-lg), inter alia, performs differently than a systemic administration in combination with the secretable vaccine protein (e.g., gp96-lg). Not wishing to be bound by theory, the effects of a cell secreting a vaccine protein alone [e.g., gp96-lg) and in combination with escalating doses of a cell secreting a T-cell costimulatory fusion protein {i.e., OX40L-lg), performs differently when compared to the vaccine protein (e.g., gp96-lg) and escalating doses of a systemic 0X40 agonist antibody.
[0007] In some embodiments, the amount of a secretable vaccine protein (e.g., gp96-lg) secretion is higher than the expression of a T cell costimulatory fusion protein, (e.g., OX40L-lg). In some embodiments, the ratio of a vaccine protein (e.g., gp96-lg) secretion to the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) is about 1 :10, 1 :25, 1 :50, 1 :100, 1 :200, 1 :300, 1 :400, 1 :500, 1 :600, 1 :700, 1 :800, 1 :900 or 1 :1000, (inclusive of all endpoints).
[0008] In some embodiments, the amount of a vaccine protein (e.g., gp96-lg) secretion is lower than the expression of a T cell costimulatory fusion protein {e.g., OX40L-lg). In some embodiments, the ratio of a vaccine protein (e.g., gp96-lg) secretion to the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) is about 10:1 , 25:1 , 50:1 , 100:1 , 200:1 , 300:1 , 400:1 , 500:1 , 600:1 , 700:1 , 800:1 , 900:1 , or 1000:1 , (inclusive of all endpoints).
[0009] In some embodiments, the amount of the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) is higher than the secretion of a vaccine protein (e.g., gp96-lg). In some embodiments, the ratio of the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) to the secretion of a vaccine protein (e.g., gp96-lg) is about 1 :10, 1 :25, 1 :50, 1 :100, 1 :200, 1 :300, 1 :400, 1 :500, 1 :600, 1 :700, 1 :800, 1 :900, or 1 :1000, (inclusive of all endpoints).
[0010] In some embodiments, the amount of the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) is lower than the secretion of a vaccine protein (e.g., gp96-lg). In some embodiments, the ratio of the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) to the secretion of a vaccine protein (e.g., gp96-lg) is about 10:1 , 25:1 , 50:1 , 100:1 , 200:1 , 300:1 , 400:1 , 500:1 , 600:1 , 700:1 , 800:1 , 900:1 , or 1000:1 , (inclusive of all endpoints).
[0011] In some embodiments, the amount of a secretable vaccine protein (e.g., gp96-lg) secretion is about the same as the expression of a T cell costimulatory fusion protein, (e.g., OX40L-lg). In some embodiments, the ratio of a vaccine protein (e.g., gp96-lg) secretion to the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) is about 1 :1. In some embodiments, the ratio of a vaccine protein (e.g., gp96-lg) secretion to the expression of a T cell costimulatory fusion protein, such as OX40L- Ig, is about 1 :1.3.
[0012] In some embodiments, the amount of a secretable vaccine protein (e.g., gp96-lg) expression is about the same as the expression of a T cell costimulatory fusion protein, (e.g., OX40L-lg). In some embodiments, the ratio of a vaccine protein (e.g., gp96-lg) expression to the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) is about 1 :1. In some embodiments, the ratio of a vaccine protein (e.g., gp96-lg) expression to the expression of a T cell costimulatory fusion protein, such as OX40L-lg, is about 1 :1 .3.
[0013] In some embodiments, the amount of the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) is about the same as the secretion of a vaccine protein (e.g., gp96-lg). In some embodiments, the ratio of the expression of a T cell costimulatory fusion protein (e.g., OX40L-lg) to the secretion of a vaccine protein (e.g., gp96-lg) is about 1 :1.
[0014] In some embodiments, the number of cells secreting a gp96-lg is higher than the number of cells secreting a OX40L-lg. In some embodiments, the ratio of the number of cells secreting a gp96-lg to the
number of cells secreting a OX40L-lg is about 1: 0.01, about 1: 0.1, about 1: 1, about 1:10, about 1:25, about 1:50, about 1:100, about 1:200, about 1:300, about 1:400, about 1:500, about 1:600, about 1:700, about 1 :800, about 1 :900 or about 1 :1000 (inclusive of all endpoints).
[0015] In some embodiments, the number of cells secreting a gp96-lg is lower than the number of cells secreting a OX40L-lg. In some embodiments, the ratio of the number of cells secreting a gp96-lg to the number of cells secreting a OX40L-lg is about 0.01: 1, about 0.1: 1, about 1:1, about 1:1.3, about 10:1, about 25:1, about 50:1, about 100:1, about 200:1, about 300:1, about 400:1, about 500:1, about 600:1, about 700:1, about 800:1, about 900:1, or about 1000:1 (inclusive of all endpoints).
[0016] In some embodiments, the expression of a gp96-lg is higher than the expression of a OX40L-lg. In some embodiments, the ratio of the expression of the gp96-lg to the expression of the OX40L-lg is about 1: 0.01, about 1: 0.1, about 1: 1, about 1:10, about 1:25, about 1:50, about 1:100, about 1:200, about 1:300, about 1:400, about 1:500, about 1:600, about 1:700, about 1:800, about 1:900 or about 1:1000 (inclusive of all endpoints).
[0017] In some embodiments, the expression of a gp96-lg is lower than the expression of a OX40L-lg. In some embodiments, the ratio of the expression of a gp96-lg to the expression of a OX40L-lg is about 0.01: 1, aboutO.1: 1, about 1:1, about 1:3, about 10:1, about 25:1, about 50:1, about 100:1, about 200:1, about 300:1, about 400:1, about 500:1, about 600:1, about 700:1, about 800:1, about 900:1, or about 1000:1 (inclusive of all endpoints).
[0018] In some embodiments, an inducible promoter can be used for inducing the expression of the vaccine protein {e.g., gp96-lg). In some embodiments, the gp96-lg is under a strong inducible promoter. In some embodiments, the gp96-lg is under an intermediate inducible promoter. In some embodiments, the gp96-lg is under a weak inducible promoter.
[0019] In some embodiments, an inducible promoter can be used for inducing the expression T cell costimulatory fusion {e.g., OX40L-lg). In some embodiments, the OX40L-lg is under a strong inducible promoter. In some embodiments, the OX40L-lg is under an intermediate inducible promoter. In some embodiments, the OX40L-lg is under a weak inducible promoter.
[0020] In some embodiments, the vaccine protein (e.g., gp96-lg) and/or T cell costimulatory fusion protein (e.g., OX40L-lg) are expressed in host cells (e.g., mammalian cells). In some embodiments, expression and/or secretion of the gp96-lg and/or OX40L-lg can be readily detected and quantified by techniques known in the art, such as, in vitro cell culturing methods or protein detection assays. In some embodiments, the protein detection assays include enzyme-linked immunosorbent assay (ELISA), immunoprecipitation, and fluorescence based methods.
[0021] In some embodiments, the amount of secreted gp96-lg by a cell is higher than the amount of
secreted OX40L-lg by a cell. In some embodiments, the ratio of the secreted gp96-lg by a cell to the secreted OX40L-lg by a cell is about 1 : 0.01 , about 1 : 0.1 , about 1 : 1 , about 1 :10, about 1 :25, about 1 :50, about 1 :100, about 1 :200, about 1 :300, about 1 :400, about 1 :500, about 1 :600, about 1 :700, about 1 :800, about 1 :900 or about 1 :1000 (inclusive of all endpoints).
[0022] In some embodiments, the amount of secreted gp96-lg by a cell is lower than the amount of secreted OX40L-lg by a cell. In some embodiments, the ratio of secreted gp96-lg by a cell to secreted OX40L-lg by a cell is about 0.01 : 1 , about 0.1 : 1 , about 1 :1 , about 1 :1.3, about 10:1 , about 25:1 , about 50:1 , about 100:1 , about 200:1 , about 300:1 , about 400:1 , about 500:1 , about 600:1 , about 700:1 , about 800:1 , about 900:1 , or about 1000:1 (inclusive of all endpoints).
[0023] In one aspect, the disclosure provides a method for treating a patient comprising administering to the patient an effective amount of a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein, wherein the patient is undergoing a treatment with a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject.
[0024] In one aspect, the disclosure provides a method for treating a patient comprising administering to the patient an effective amount of a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein, wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject, and wherein the patient is undergoing a treatment with a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein.
[0025] In one aspect, the disclosure provides a method for treating a patient comprising administering to the patient an effective amount of (a) a first cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and (b) a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject.
[0026] In some embodiments, the secretable vaccine protein is a secretable gp96-lg fusion protein which optionally lacks the gp96 KDEL (SEQ ID NO:3) sequence. In some embodiments, the Ig tag in the gp96-lg fusion protein comprises the Fc region of human lgG1 , lgG2, lgG3, lgG4, IgM, IgA, or IgE.
[0027] In some embodiments, the T cell costimulatory fusion protein is OX40L-lg, or a portion thereof that binds to 0X40. In some embodiments, the T cell costimulatory fusion protein is ICOSL-lg, or a portion thereof that binds to ICOS. In some embodiments, the T cell costimulatory fusion protein is 4-1 BBL-lg, or
a portion thereof that binds to 4-1 BBR. In some embodiments, the T cell costimulatory fusion protein is TL1 A-lg, or a portion thereof that binds to TNFRSF25. In some embodiments, the T cell costimulatory fusion protein is GITRL-lg, or a portion thereof that binds to GITR. In some embodiments, the T cell costimulatory fusion protein is CD40L-lg, or a portion thereof that binds to CD40. In some embodiments, the T cell costimulatory fusion protein is CD70-lg, or a portion thereof that binds to CD27. In some embodiments, the Ig tag in the T cell costimulatory fusion protein comprises the Fc region of human lgG1 , lgG2, lgG3, lgG4, IgM, IgA, or IgE.
[0028] In some embodiments, the expression vector is incorporated into a virus or virus-like particle. In some embodiments, the expression vector is incorporated into a human tumor cell. In some embodiments, the patient is a human cancer patient. In some embodiments, administration to the human patient increases the activation or proliferation of tumor antigen specific T cells in the patient.
[0029] In some embodiments, the activation or proliferation of tumor antigen specific T cells in the patient is increased by at least 25 percent as compared to the level of activation or proliferation of tumor antigen specific T cells in the patient prior to the administration.
[0030] In some embodiments, administration is in combination with an agent that inhibits immunosuppressive molecules produced by tumor cells. In some embodiments, the agent is an antibody against PD-1. In some embodiments, the antibody against PD-1 is selected from Nivolumab, Pembrolizumab, Pidilizumab, Cemiplimab, AGEN2034, AMP-224, AMP-514, PDR001.
[0031] In some embodiments, the patient is a human with an acute or chronic infection. In some embodiments, the acute or chronic infection is an infection by hepatitis C virus, hepatitis B virus, human immunodeficiency virus, or malaria.
[0032] In some embodiments, administration to the human patient stimulates the activation or proliferation of pathogenic antigen specific T cells.
[0033] In some embodiments, the T cell costimulatory molecule enhances the activation of antigen- specific T cells in the subject to a greater level than gp96-lg vaccination alone.
BRIEF DESCRIPTION OF THE DRAWINGS
[0034] FIG. 1 is a plasmid vector map for pcDNA3.4 OX40L-lg.
[0035] FIG. 2 is a graph showing the activation of human 0X40 receptor in Jurkat cells by mouse and human OX40L.
[0036] FIG. 3 is an image showing mouse HS-110 (B16F10-OVA-gp96) and mouse HS-130 (B16F10- OVA- OX40L) immunization over a dose-range to correlate CD8+T-cell expansion to tumor growth delay.
[0037] FIG. 4 shows flow cytometry diagrams, dot plots and gating strategy for Day 7 after a primary vaccination (peripheral blood). Recipient mice were injected with a mouse mHS-110 at a constant dose of 1 million cells (290 ng of gp96-lg) with different ratios of mHS-130 (0.1 , 0.3, 1 , 3, 10). A ratio of 1 to 1 is 290 ng of gp96 (1 million mHS-110 cells) to 290 ng of OX40L. FIG. 4 shows a flow cytometry gating strategy using FlowJo Version 10 (2018), by gating in the blood on singlets and CD3+ T cells, then CD8+ OT-I GFP+ T cells. Sample analysis was taken on day 7 and numbers in representative dot plots indicate percentages of CD8+ OT-I GFP+ positive cells within the gated population. Plots show an individual, representative mouse that illustrates peak expansion for the day selected.
[0038] FIG. 5 shows flow cytometry diagrams, dot plots and gating strategy for Day 21 after a boost vaccination (peripheral blood). Recipient mice were injected with a mouse mHS-110 at a constant dose of 1 million cells (290 ng of gp96-lg) with different ratios of mHS-130 (0.1 , 0.3, 1 , 3, 10). A ratio of 1 to 1 is 290 ng of gp96 (1 million mHS-110 cells) to 290 ng of OX40L. FIG. 5 shows a flow cytometry gating strategy using FlowJo Version 10 (2018), by gating in the blood on singlets and CD3+ T cells, then CD8+ OT-I GFP+ T cells. Sample analysis was taken on day 21 after a boost immunization that was given on day 14, and numbers in the representative dot plots indicate percentages of CD8+ OT-I GFP+ positive cells within the gated population. Plots show an individual, representative mouse that illustrates peak expansion for the day selected.
[0039] FIG. 6A and FIG. 6B are graphs showing the percent of OT-I CD8+ T-cells after primary and secondary vaccination with a set dose mHS-110 with different ratios of mHS-130 before and after tumor challenge (peripheral blood). Recipient mice were injected with a mHS-110 at a constant dose of 1 million cells (290 ng of gp96-lg) with different ratios of mHS130. After vaccination, OT-I GFP+ CD8+T cells were analyzed in the blood on days 0-53 days post-vaccination. Then mice were boosted on day 14 with the same ratios of mHS110 and mHS130 as in the primary phase, and OT-I GFP+ CD8+ T cells were analyzed in the blood on days 17, 19, 21 , 24, 28, 33, 38, 41 days post-challenge. Data represent mean total numbers ± SEM from n = 5 mice. *p < 0.05 **p < 0.01 (mHS-110 Only versus different ratios of mHS-130). (FIG. 6A): Line graph without overlay; (FIG. 6B): Line graph with mouse outliers removed out to day 41 only.
[0040] FIG. 7A, FIG. 7B, FIG. 7C, FIG. 7D, and FIG. 7E are graphs showing the day-54, End-of-Study, endogenous response to vaccination. FIG. 7A is a graph showing an End-of study, day-54, endogenous spleen response to vaccination, percent, gated FSC-H by FSC-A as to live/dead gate for doublets, then gated on CD45 by SSC then CD3+ CD8+ double positive cells. FIG. 7A shows mean ± SEM, *p<0.05 compared to mHS-110 via the non-parameteric statistical test, Mann-Whitney. FIG. 7B is a flow cytometry diagram and FIG. 7C shows a graph of End-of study, day-54, endogenous spleen response to
vaccination and ex vivo stimulation with pg100 peptide for intracellular cytokine staining. Percent shown in graph. Events gated on FSC-H by FSC-A as to live/dead gate for doublets, then gated on CD45 by SSC then IFN-g CD8+ double positive cells. FIG. 7C is a graph showing mean ± SEM, *p<0.05 compared to mHS-110 via the non-parameteric statistical test, Mann-Whitney;‘ns’ denotes p>0.05, not significant. FIG. 7D are graphs showing endogenous spleen immune response measured by IFN-gamma ELISPOT. FIG. 7D shows graphs having mean ± SEM IFN-g spots per million splenocytes, **p<0.01 , *p<0.05 compared to mHS-1 10 via the non-parameteric statistical test, Mann-Whitney;‘ns’ denotes p>0.05, not significant. Far right shows representative ELISPOT well with machine counts. Background (media alone) wells not subtracted from graphed data sets. Positive control wells worked (data not shown). FIG. 7E is a graph showing End-of study, day-54, endogenous response to vaccination, percent, gated FSC-H by FSC-A as to live/dead gate for doublets, then gated on CD45 by SSC then CD3+ CD4+ double positive cells. FIG. 7E shows mean ± SEM, **p<0.01 compared to mHS-1 10 via the non-parameteric statistical test, Mann-Whitney.
[0041] FIG. 8A is a flow cytometry diagram and FIG. 8B is a graph showing CD8+ Tumor Infiltrating Lymphocytes (TILs). FIG. 8A shows graphs of the End-of study, day-54, endogenous TIL response to vaccination, percent, gated FSC-H by FSC-A to as live/dead gate for doublets, then gated on CD45 by SSC then CD3+ CD8+ double positive cells. The MACS Miltenyl Biotec tumor dissociation kit was used for this procedure (cat# 130-096-730). FIG. 8B is a graph showing mean ± SEM, *p<0.05 compared to mHS-1 10 via the non-parameteric statistical test, Mann-Whitney.
[0042] FIG. 9 is a graph showing End of study, final tumor mass, in grams, of individual mice. Tumor mass (wet weight) were weighed using a milligram sensitive scale for each animal. FIG. 9 shows mean ± SEM. Statistics performed were non-parametric Mann-Whitney, ‘ns’ designates a non-significant (p>0.05) value, *p<0.05; **p<0.01.
[0043] FIG.10A and FIG. 10B shows tumor volumes over time, tumor mean size, and individual plots for individual animals. FIG. 10A is a graph showing mean ± SEM for all tumor volumes over time. FIG. 10B is a graph showing means for individual animals for each measured timepoint. Tumor implantation, melanoma B16F10 cells were harvested and resuspended at a concentration of 5x105 cells/1 OOpl in a volume of 80mI HBSS and 20pl Matrigel. C57BL/6 mice were subcutaneously injected with 100 mI of B16F10 cells (5 x 105cells/mouse) on the inner abdomen . The tumor size was measured and documented every 3 days with a caliper, starting on day 7, and calculated using the formula (A * B) (A as the largest and B as the smallest diameter of tumor). Tumor growth was documented as standard error mean. To record the survival of the tumor-bearing mice, either natural death or a tumor volume greater than 450 mm2 leading to death was counted as death. Each experimental group included five animals. Statistics
performed for FIG. 10A was a 2-way ANOVA (shown below FIG. 10A), p<0.05 is considered significantly different vs. mHS-1 10 group alone.
[0044] FIG. HA and FIG. 11 B are graphs showing the percent CD8+ OT-1 + T-cells on day 54 (spleen), and the flow plot gating strategy. FIG. 11 A is a graph showing the End-of study, day-54, endogenous response to vaccination, percent, gated FSC-H by FSC-A as to live/dead gate for doublets, then gated on CD45 by SSC then GFP-OT-1 CD8+ double positive cells. FIG. 11 B is a graph showing mean ± SEM, *p<0.05 compared to mHS-110 via the non-parametric statistical test, Mann-Whitney.
[0045] FIG. 12A and FIG. 12B are graphs showing the percent CD8+ PD-1 + T-cells on day 54 (spleen), and the flow plot gating strategy. FIG. 12A is a graph showing End-of study, day-54, endogenous response to vaccination, percent, gated FSC-H by FSC-A as to live/dead gate for doublets, then gated on CD3 by SSC, then PD-1 + CD8+ double positive cells. FIG. 12B is a graph showing mean ± SEM, *p<0.05 compared to mHS-110 via the non-parameteric statistical test, Mann-Whitney.
[0046] FIG. 13 is a non-limiting schematic of the design of the study of gp96-lg (mHS-110, B16F10- OVA-gp96) to OX40L-lg (mHS-130, B16F10-OVA- OX40L) dose ratios, to correlate CD8+ T-cell expansion to tumor growth delay.
[0047] FIG. 14 are graphs illustrating anti-tumor CD8+ OT-I T cell expansion in the peripheral blood with prime and boost Immunization of different ratios and dose combinations of mHS-1 10 and mHS-130, in the study of FIG. 13. Recipient mice were injected with mHS-1 10 and mHS-130 at different ratios and doses of gp96-lg to OX40L-lg. OT-I GFP+ CD8+ T cells were analyzed in the blood on days 0-54 days post-vaccination. Mice were boosted on day 14 with the same ratios of mHS-110 and mHS-130 as in the primary phase, and OT-I GFP+ CD8+ T cells were analyzed in the blood days post-challenge. Data represent mean percent ± SEM.
[0048] FIGS. 15A to 15D illustrate flow cytometry gating strategy and cellular expansion of CD8+ OT-I T-cells, over time, with mHS-110/130 immunization in the study of FIG. 13. FIG. 15A are graphs illustrating flow cytometry gating strategy of CD8+ OT-I T-cells, over time, for the tested ratios and mPIS- 110/130 immunization doses. FIG. 15B are bar charts illustrating expansion of CD8+ OT -I T-cells on days 7 and 17. FIG. 15C are bar charts illustrating expansion of CD8+ OT-I T-cells on days 19, 21 , 24, 26, 28, 33, 38, and 41 . FIG. 15D are bar charts illustrating expansion of CD8+ OT-I T-cells on days 45, 48, and 54. Data represent mean percent ± SEM. Statistical analysis was Mann-Whitney, *p<0.05, **p<0.01 , ***p<0.001 ;‘ns’ denotes p>0.05 or‘not significant’.
[0049] FIGS. 16, 17 and 18 illustrate percent of T-cells in the peripheral blood for SLECs, MPECs, Activated/C D44hi CD8+ endogenous and exogenous (OT-I), T-cells, on day 7 of the study of FIG. 13. FIG. 16 are graphs illustrating flow cytometry gating strategy for MPECs and SLECs. FIG. 17 are bar
charts illustrating MPECs and SLEC for endogenous CD8+ T-cells. FIG. 18 bar charts illustrating percent of CD44hi endogenous CD8+ T-cells (% CD8+ CD44+ T cells).
[0050] FIGS. 19 and 20 are graphs illustrating tumor growth delay/inhibition over time, in the study of FIG. 13. FIG. 19 shows tumor diameter (in mm3) for each of the dose ratio groups, for days 0-28, as mean ± SEM grouped tumor diameter growth curves. FIG. 20 are bar charts (left panel) illustrating tumor weights (in grams) as mean ± SEM, and scatter plots (right panel) illustrating individual tumors (in grams) as mean ± SEM. Statistical analysis performed was Mann-Whitney, *p<0.05, **p<0.01 ;‘ns’ denotes p>0.05 or‘not significant'.
[0051] FIG. 21 are bar charts illustrating percent of CD3+ CD8+ tetramer-TRP2+ T-cells in the spleen on day 55 of the study of FIG. 13. Graphed values for gated samples are shown, and represent mean percent ± SEM. Statistical analysis performed was Mann-Whitney, *p<0.05, **p<0.01 ;‘ns’ denotes p>0.05 or‘not significant'.
[0052] FIG. 22 are bar charts illustrating percent of CD3+ CD8+ eGFP/OT-1 + T-cells in the spleen and blood on day 55 of the study of FIG. 13. CD8+eGFP/OT-1 + T-cells gated for blood and spleen are shown, and represent mean percent ± SEM. Statistical analysis performed was Mann-Whitney.
[0053] FIG. 23 are bar charts illustrating splenocytes phenotypes on day 55. Data shows percent of CD3+ CD4+ PD-1 + T cells in the spleen on day 55 of the study of FIG. 13. Data represent mean percent ± SEM. Statistical analysis performed was Mann-Whitney,‘ns’ denotes p>0.05 or‘not significant'.
[0054] FIG. 24 are bar charts illustrating percent of CD3+ CD4+ CD44/CD62L central memory T-cells in the spleen on day 55 of the study of FIG. 13. Data represent mean percent ± SEM. Statistical analysis performed was Mann-Whitney, *p<0.05, **p<0.01.
[0055] FIG. 25 are bar charts illustrating tumor infiltrating lymphocytes (TILs) phenotypes. CD8+ TILs (% CD8+ CD3+ T-cells) are shown on day 55 of the study of FIG. 13. Data represent mean percent ± SEM. Statistical analysis performed was Mann-Whitney, *p<0.05,‘ns’ denotes p>0.05 or‘not significant'.
[0056] FIG. 26 are bar charts illustrating tumor infiltrating lymphocytes (TILs) phenotypes. CD4+ TILs (% CD4+ CD3+ T cells) are shown on day 55 of the study of FIG. 13. Data represent mean percent ± SEM from. Statistical analysis performed was Mann-Whitney, *p<0.05; ‘ns’ denotes p>0.05 or‘not significant'.
DETAILED DESCRIPTION OF THE DISCLOSURE
[0057] The various secretable proteins, i.e., vaccine proteins as described herein, can be used to stimulate an immune response in vivo. For example, secretable heat-shock protein gp96-lg based allogeneic cellular vaccines can achieve high-frequency polyclonal CD8+ T cell responses to femto-molar
concentrations of tumor antigens through antigen cross-priming in vivo ( Oizumi et al, J Immunol 2007, 179(4) :2310-2317) . Multiple immunosuppressive mechanisms elaborated by established tumors can dampen the activity of this vaccine approach, however. In combination immunotherapy for patients with advanced disease, a systematic comparison of PD-1 , PD-L1 , CTLA-4, and LAG-3 blocking antibodies in mouse models of long-established B16-F10 melanoma demonstrated superior combination between gp96-lg vaccination and PD-1 blockade as compared to other checkpoints. Synergistic anti-tumor benefits may result from triple combinations of gp96-lg vaccination, PD-1 blockade, and T cell costimulation using one or of an agonist of 0X40 {e.g., an 0X40 ligand-lg (OX40L-lg) fusion, or a fragment thereof that binds 0X40), an agonist of inducible T-cell costimulator (ICOS) {e.g., an ICOS ligand-lg (ICOSL-lg) fusion, or a fragment thereof that binds ICOS), an agonist of CD40 (e.g., a CD40L- Ig fusion protein, or fragment thereof), an agonist of CD27 (e.g., a CD70-lg fusion protein or fragment thereof), an agonist of 4-1 BB (e.g., a 4-1 BB ligand-lg (4-1 BBL-lg) fusion, or a fragment thereof that binds 4-1 BB), an agonist of TNFRSF25 (e.g., a TL1 A-lg fusion, or a fragment thereof that binds TNFRSF25), or an agonist of glucocorticoid-induced tumor necrosis factor receptor (GITR) (e.g., a GITR ligand-lg (GITRL-lg) fusion, or a fragment thereof that binds GITF). Secretion of gp96-lg and these costimulatory fusion proteins by allogeneic cell lines, enhances activation of antigen-specific CD8+ T cells. Notwithstanding any theory, the effect of a locally secreted T-cell costimulatory fusion protein (i.e., OX40L-lg), inter alia, performs differently than a systemic administration in combination with the secretable vaccine protein (e.g., gp96-lg).
Vaccine Proteins
[0058] Vaccine proteins can induce immune responses that find use in the present invention. In some embodiments, the disclosure provides a cell based therapy comprising a first cell comprising an expression vector comprising, a nucleotide sequence that encode a secretable vaccine protein and a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein. Compositions useful in the cell based therapy of the present invention are also provided. In various embodiments, such compositions are utilized in methods of treating subjects to stimulate immune responses in the subject including enhancing the activation of antigen-specific T cells in the subject. The present compositions find use in the treatment of various diseases including cancer.
[0059] The heat shock protein (hsp) gp96, localized in the endoplasmic reticulum (ER), serves as a chaperone for peptides on their way to MHC class I and II molecules. Gp96 obtained from tumor cells and used as a vaccine can induce specific tumor immunity, presumably through the transport of tumor- specific peptides to antigen-presenting cells (APCs) (J Immunol 1999, 163(10):5178-5182). For example,
gp96-associated peptides are cross-presented to CD8 cells by dendritic cells (DCs).
[0060] A vaccination system was developed for antitumor therapy by transfecting a gp96-lg G1-Fc fusion protein into tumor cells, resulting in secretion of gp96-lg in complex with chaperoned tumor peptides (see, J Immunother 2008, 31 (4):394-401 , and references cited therein). Parenteral administration of gp96-lg secreting tumor cells triggers robust, antigen-specific CD8 cytotoxic T lymphocyte (CTL) expansion, combined with activation of the innate immune system. Tumor-secreted gp96 causes the recruitment of DCs and natural killer (NK) cells to the site of gp96 secretion, and mediates DC activation. Further, the endocytic uptake of gp96 and its chaperoned peptides triggers peptide cross presentation via major MHC class I, as well as strong, cognate CD8 activation independent of CD4 cells.
[0061] The cell based therapy provided herein involve a first nucleotide sequence that encodes a gp96- Ig fusion protein. The coding region of human gp96 is 2,412 bases in length (SEQ ID NO:1), and encodes an 803 amino acid protein (SEQ ID NO:2) that includes a 21 amino acid signal peptide at the amino terminus, a potential transmembrane region rich in hydrophobic residues, and an ER retention peptide sequence at the carboxyl terminus (GENBANK® Accession No. X15187; see, Maki et al., Proc Natl Acad Sci USA 1990, 87:5658-5562). The DNA and protein sequences of human gp96 follow:
atgagggccctgtgggtgctgggcctctgctgcgtcctgctgaccttcgggtcggtcagagctgacgatgaagttgatgtg
gatggtacagtagaagaggatctgggtaaaagtagagaaggatcaaggacggatgatgaagtagtacagagagag
gaagaagctattcagttggatggattaaatgcatcacaaataagagaacttagagagaagtcggaaaagtttgccttcc
aagccgaagttaacagaatgatgaaacttatcatcaattcattgtataaaaataaagagattttcctgagagaactgatttc
aaatgcttctgatgctttagataagataaggctaatatcactgactgatgaaaatgctctttctggaaatgaggaactaaca
gtcaaaattaagtgtgataaggagaagaacctgctgcatgtcacagacaccggtgtaggaatgaccagagaagagttg
gttaaaaaccttggtaccatagccaaatctgggacaagcgagtttttaaacaaaatgactgaagcacaggaagatggc
cagtcaacttctgaattgattggccagtttggtgtcggtttctattccgccttccttgtagcagataaggttattgtcacttcaaaa
cacaacaacgatacccagcacatctgggagtctgactccaatgaattttctgtaattgctgacccaagaggaaacactct
aggacggggaacgacaattacccttgtcttaaaagaagaagcatctgattaccttgaattggatacaattaaaaatctcgt
caaaaaatattcacagttcataaactttcctatttatgtatggagcagcaagactgaaactgttgaggagcccatggagga
agaagaagcagccaaagaagagaaagaagaatctgatgatgaagctgcagtagaggaagaagaagaagaaaa
gaaaccaaagactaaaaaagttgaaaaaactgtctgggactgggaacttatgaatgatatcaaaccaatatggcaga
gaccatcaaaagaagtagaagaagatgaatacaaagctttctacaaatcattttcaaaggaaagtgatgaccccatgg
cttatattcactttactgctgaaggggaagttaccttcaaatcaattttatttgtacccacatctgctccacgtggtctgtttgacg
aatatggatctaaaaagagcgattacattaagctctatgtgcgccgtgtattcatcacagacgacttccatgatatgatgcct
aaatacctcaattttgtcaagggtgtggtggactcagatgatctccccttgaatgtttcccgcgagactcttcagcaacataa
actgcttaaggtgattaggaagaagcttgttcgtaaaacgctggacatgatcaagaagattgctgatgataaatacaatg
atactttttggaaagaatttggtaccaacatcaagcttggtgtgattgaagaccactcgaatcgaacacgtcttgctaaactt
cttaggttccagtcttctcatcatccaactgacattactagcctagaccagtatgtggaaagaatgaaggaaaaacaaga
caaaatctacttcatggctgggtccagcagaaaagaggctgaatcttctccatttgttgagcgacttctgaaaaagggctat
gaagttatttacctcacagaacctgtggatgaatactgtattcaggcccttcccgaatttgatgggaagaggttccagaatg
ttgccaaggaaggagtgaagttcgatgaaagtgagaaaactaaggagagtcgtgaagcagttgagaaagaatttgag
cctctgctgaattggatgaaagataaagcccttaaggacaagattgaaaaggctgtggtgtctcagcgcctgacagaat
ctccgtgtgctttggtggccagccagtacggatggtctggcaacatggagagaatcatgaaagcacaagcgtaccaaa
cgggcaaggacatctctacaaattactatgcgagtcagaagaaaacatttgaaattaatcccagacacccgctgatcag
agacatgcttcgacgaattaaggaagatgaagatgataaaacagttttggatcttgctgtggttttgtttgaaacagcaacg
cttcggtcagggtatcttttaccagacactaaagcatatggagatagaatagaaagaatgcttcgcctcagtttgaacattg
accctgatgcaaaggtggaagaagagcccgaagaagaacctgaagagacagcagaagacacaacagaagaca
cagagcaagacgaagatgaagaaatggatgtgggaacagatgaagaagaagaaacagcaaaggaatctacagct
gaaaaagatgaattgtaa (SEQ ID N0:1)
MRALWVLGLCCVLLTFGSVRADDEVDVDGTVEEDLGKSREGSRTDDEVVQREEEAI
QLDGLNASQIRELREKSEKFAFQAEVNRMMKLIINSLYKNKEIFLRELISNASDALDKIR
LISLTDENALSGNEELTVKIKCDKEKNLLHVTDTGVGMTREELVKNLGTIAKSGTSEFL
NKMTEAQEDGQSTSELIGQFGVGFYSAFLVADKVIVTSKHNNDTQHIWESDSNEFSVI
ADPRGNTLGRGTTITLVLKEEASDYLELDTIKNLVKKYSQFINFPIYVWSSKTETVEEP
MEEEEAAKEEKEESDDEAAVEEEEEEKKPKTKKVEKTVWDWELMNDIKPIWQRPSK
EVEEDEYKAFYKSFSKESDDPMAYIHFTAEGEVTFKSILFVPTSAPRGLFDEYGSKKS
DYIKLYVRRVFITDDFHDMMPKYLNFVKGVVDSDDLPLNVSRETLQQHKLLKVIRKKLV
RKTLDMIKKIADDKYNDTFWKEFGTNIKLGVIEDHSNRTRLAKLLRFQSSHHPTDITSL
DQYVERMKEKQDKIYFMAGSSRKEAESSPFVERLLKKGYEVIYLTEPVDEYCIQALPE
FDGKRFQNVAKEGVKFDESEKTKESREAVEKEFEPLLNWMKDKALKDKIEKAWSQR
LTESPCALVASQYGWSGNMERIMKAQAYQTGKDISTNYYASQKKTFEINPRHPLIRD
MLRRIKEDEDDKTVLDLAVVLFETATLRSGYLLPDTKAYGDRIERMLRLSLNIDPDAKV
EEEPEEEPEETAEDTTEDTEQDEDEEMDVGTDEEEETAKESTAEKDEL (SEQ ID
N0:2).
[0062] A nucleic acid encoding a gp96-lg fusion sequence can be produced using the methods described in U.S. Patent No. 8,685,384, which is incorporated herein by reference in its entirety. In some embodiments, the gp96 portion of a gp96-lg fusion protein can contain all or a portion of a wild type gp96 sequence {e.g., the human sequence set forth in SEQ ID NO:2). For example, a secretable gp96-lg fusion protein can include the first 799 amino acids of SEQ ID NO:2, such that it lacks the C-terminal KDEL (SEQ ID NO:3) sequence. Alternatively, the gp96 portion of the fusion protein can have an amino acid sequence that contains one or more substitutions, deletions, or additions as compared to the first 799 amino acids of the wild type gp96 sequence, such that it has at least 90% {e.g., at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity to the wild type polypeptide.
[0063] As used throughout this disclosure, the percent sequence identity between a particular nucleic acid or amino acid sequence and a sequence referenced by a particular sequence identification number is determined as follows. First, a nucleic acid or amino acid sequence is compared to the sequence set
forth in a particular sequence identification number using the BLAST 2 Sequences (BI2seq) program from the stand-alone version of BLASTZ containing BLASTN version 2.0.14 and BLASTP version 2.0.14. This stand-alone version of BLASTZ can be obtained online at fr.com/blast or at ncbi.nlm.nih.gov. Instructions explaining how to use the BI2seq program can be found in the readme file accompanying BLASTZ. BI2seq performs a comparison between two sequences using either the BLASTN or BLASTP algorithm. BLASTN is used to compare nucleic acid sequences, while BLASTP is used to compare amino acid sequences. To compare two nucleic acid sequences, the options are set as follows: -i is set to a file containing the first nucleic acid sequence to be compared {e.g., C:\seq1 .bet); -j is set to a file containing the second nucleic acid sequence to be compared {e.g., C:\seq2.txt); -p is set to blastn; -o is set to any desired file name (e.g., C:\output.txt); -q is set to -1 ; -r is set to 2; and all other options are left at their default setting. For example, the following command can be used to generate an output file containing a comparison between two sequences: C:\BI2seq -i c:\seq1 .txt -j c:\seq2.txt -p blastn -o c:\output.txt -q -1 -r 2. To compare two amino acid sequences, the options of BI2seq are set as follows: -i is set to a file containing the first amino acid sequence to be compared (e.g., C:\seq1 .txt); -j is set to a file containing the second amino acid sequence to be compared (e.g., C:\seq2.txt); -p is set to blastp; -o is set to any desired file name (e.g., C:\output.txt); and all other options are left at their default setting. For example, the following command can be used to generate an output file containing a comparison between two amino acid sequences: C:\BI2seq -i c:\seq1.txt -j c:\seq2.txt -p blastp -o c:\output.txt. If the two compared sequences share homology, then the designated output file will present those regions of homology as aligned sequences. If the two compared sequences do not share homology, then the designated output file will not present aligned sequences.
[0064] Once aligned, the number of matches is determined by counting the number of positions where an identical nucleotide or amino acid residue is presented in both sequences. The percent sequence identity is determined by dividing the number of matches either by the length of the sequence set forth in the identified sequence (e.g., SEQ ID NO:1 ), or by an articulated length (e.g., 100 consecutive nucleotides or amino acid residues from a sequence set forth in an identified sequence), followed by multiplying the resulting value by 100. For example, a nucleic acid sequence that has 2,200 matches when aligned with the sequence set forth in SEQ ID NO:1 is 91 .2 percent identical to the sequence set forth in SEQ ID NO:1 (i.e., 2,000 ÷ 2,412 x 100 = 91.2). It is noted that the percent sequence identity value is rounded to the nearest tenth. For example, 75.11 , 75.12, 75.13, and 75.14 is rounded down to 75.1 , while 75.15, 75.16, 75.17, 75.18, and 75.19 is rounded up to 75.2. It also is noted that the length value will always be an integer.
[0065] Thus, in some embodiments, the gp96 portion of nucleic acid encoding a gp96-lg fusion
polypeptide can encode an amino acid sequence that differs from the wild type gp96 polypeptide at one or more amino acid positions, such that it contains one or more conservative substitutions, non conservative substitutions, splice variants, isoforms, homologues from other species, and polymorphisms.
[0066] As defined herein, a "conservative substitution” denotes the replacement of an amino acid residue by another, biologically similar, residue. Typically, biological similarity, as referred to above, reflects substitutions on the wild type sequence with conserved amino acids. For example, conservative amino acid substitutions would be expected to have little or no effect on biological activity, particularly if they represent less than 10% of the total number of residues in the polypeptide or protein. Conservative substitutions may be made, for instance, on the basis of similarity in polarity, charge, size, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the amino acid residues involved. The 20 naturally occurring amino acids can be grouped into the following six standard amino acid groups: (1) hydrophobic: Met, Ala, Val, Leu, lie; (2) neutral hydrophilic: Cys, Ser, Thr; Asn, Gin; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe. Accordingly, conservative substitutions may be effected by exchanging an amino acid by another amino acid listed within the same group of the six standard amino acid groups shown above. For example, the exchange of Asp by Glu retains one negative charge in the so modified polypeptide. In addition, glycine and proline may be substituted for one another based on their ability to disrupt a-helices. Additional examples of conserved amino acid substitutions, include, without limitation, the substitution of one hydrophobic residue for another, such as isoleucine, valine, leucine, or methionine, or the substitution of one polar residue for another, such as the substitution of arginine for lysine, glutamic for aspartic acid, or glutamine for asparagine, and the like. The term "conservative substitution” also includes the use of a substituted amino acid residue in place of an un-substituted parent amino acid residue, provided that antibodies raised to the substituted polypeptide also immunoreact with the un-substituted polypeptide.
[0067] As used herein, "non-conservative substitutions” are defined as exchanges of an amino acid by another amino acid listed in a different group of the six standard amino acid groups (1) to (6) shown above.
[0068] In various embodiments, the substitutions may also include non-classical amino acids {e.g., selenocysteine, pyrrolysine, N-formylmethionine b-alanine, GABA and d-Aminolevulinic acid, 4- aminobenzoic acid (PABA), D-isomers of the common amino acids, 2,4-diaminobutyric acid, a-amino isobutyric acid, 4-aminobutyric acid, Abu, 2-amino butyric acid, y-Abu, e-Ahx, 6-amino hexanoic acid, Aib, 2-amino isobutyric acid, 3-amino propionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosme, citrulline, homocitrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylglycine,
cyclohexylalanine, b-alanine, fluoro-amino acids, designer amino acids such as b methyl amino acids, C a-methyl amino acids, N a-methyl amino acids, and amino acid analogs in general).
[0069] Mutations may also be made to the nucleotide sequences of the present fusion proteins by reference to the genetic code, including taking into account codon degeneracy.
[0070] The Ig portion ("tag”) of a gp96-lg fusion protein can contain, for example, a non-variable portion of an immunoglobulin molecule {e.g., an lgG1 , lgG2, lgG3, lgG4, IgM, IgA, or IgE molecule). Typically, such portions contain at least functional CH2 and CH3 domains of the constant region of an immunoglobulin heavy chain. Fusions also can be made using the carboxyl terminus of the Fc portion of a constant domain, or a region immediately amino-terminal to the CH1 of the heavy or light chain. The Ig tag can be from a mammalian {e.g., human, mouse, monkey, or rat) immunoglobulin, but human immunoglobulin can be particularly useful when the gp96-lg fusion is intended for in vivo use for humans.
[0071] DNAs encoding immunoglobulin light or heavy chain constant regions are known or readily available from cDNA libraries. See, for example, Adams et al., Biochemistry 1980, 19:271 1-2719; Gough et al., Biochemistry 1980 19:2702-2710; Dolby et al., Proc Natl Acad Sci USA 1980, 77:6027-6031 ; Rice et al., Proc Natl Acad Sci USA 1982, 79:7862-7865; Falkner et al., Nature 1982, 298:286-288; and Morrison et al., Ann Rev Immunol 1984, 2:239-256. Since many immunological reagents and labeling systems are available for the detection of immunoglobulins, gp96-lg fusion proteins can readily be detected and quantified by a variety of immunological techniques known in the art, such as enzyme- linked immunosorbent assay (ELISA), immunoprecipitation, and fluorescence activated cell sorting (FACS). Similarly, if the peptide tag is an epitope with readily available antibodies, such reagents can be used with the techniques mentioned above to detect, quantitate, and isolate gp96-lg fusions.
[0072] In various embodiments, the gp96-lg fusion protein and/or the costimulatory molecule fusions, comprises a linker. In various embodiments, the linker may be derived from naturally-occurring multi- domain proteins or are empirical linkers as described, for example, in Chichili et al., (2013), Protein Sci. 22(2):153-167, Chen et al., (2013), Adv Drug Deliv Rev. 65(10):1357-1369, the entire contents of which are hereby incorporated by reference. In some embodiments, the linker may be designed using linker designing databases and computer programs such as those described in Chen et al., (2013), Adv Drug Deliv Rev. 65(10):1357-1369 and Crasto et. al., (2000), Protein Eng. 13(5):309-312, the entire contents of which are hereby incorporated by reference.
[0073] In some embodiments, the linker is a synthetic linker such as PEG.
[0074] In other embodiments, the linker is a polypeptide. In some embodiments, the linker is less than about 100 amino acids long. For example, the linker may be less than about 100, about 95, about 90, about 85, about 80, about 75, about 70, about 65, about 60, about 55, about 50, about 45, about 40,
about 35, about 30, about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 12, about 11 , about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, or about 2 amino acids long. In some embodiments, the linker is flexible. In another embodiment, the linker is rigid. In various embodiments, the linker is substantially comprised of glycine and serine residues (e.g. about 30%, or about 40%, or about 50%, or about 60%, or about 70%, or about 80%, or about 90%, or about 95%, or about 97% glycines and serines).
[0075] In various embodiments, the linker is a hinge region of an antibody {e.g., of IgG, IgA, IgD, and IgE, inclusive of subclasses (e.g. lgG1 , lgG2, lgG3, and lgG4, and lgA1 and lgA2)). The hinge region, found in IgG, IgA, IgD, and IgE class antibodies, acts as a flexible spacer, allowing the Fab portion to move freely in space. In contrast to the constant regions, the hinge domains are structurally diverse, varying in both sequence and length among immunoglobulin classes and subclasses. For example, the length and flexibility of the hinge region varies among the IgG subclasses. The hinge region of lgG1 encompasses amino acids 216-231 and, because it is freely flexible, the Fab fragments can rotate about their axes of symmetry and move within a sphere centered at the first of two inter-heavy chain disulfide bridges. lgG2 has a shorter hinge than lgG1 , with 12 amino acid residues and four disulfide bridges. The hinge region of lgG2 lacks a glycine residue, is relatively short, and contains a rigid poly-proline double helix, stabilized by extra inter-heavy chain disulfide bridges. These properties restrict the flexibility of the lgG2 molecule. lgG3 differs from the other subclasses by its unique extended hinge region (about four times as long as the lgG1 hinge), containing 62 amino acids (including 21 prolines and 11 cysteines), forming an inflexible poly-proline double helix. In lgG3, the Fab fragments are relatively far away from the Fc fragment, giving the molecule a greater flexibility. The elongated hinge in lgG3 is also responsible for its higher molecular weight compared to the other subclasses. The hinge region of lgG4 is shorter than that of lgG1 and its flexibility is intermediate between that of lgG1 and lgG2. The flexibility of the hinge regions reportedly decreases in the order lgG3>lgG1 >lgG4>lgG2.
[0076] Additional illustrative linkers include, but are not limited to, linkers having the sequence LE, GGGGS (SEQ ID NO:14), (GGGGS)n (n=1-4) (SEQ ID NO: 15), (Gly)8 (SEQ ID NO:16), (Gly)6 (SEQ ID NO: 17), (EAAAK)n (n=1-3) (SEQ ID NO: 18), A(EAAAK)nA (n = 2-5) (SEQ ID NO: 19), AEAAAKEAAAKA (SEQ ID NO: 20), A(EAAAK)4ALEA(EAAAK)4A (SEQ ID NO: 21), PAPAP (SEQ ID NO: 22), KESGSVSSEQLAQFRSLD (SEQ ID NO: 23), EGKSSGSGSESKST(SEQ ID NO: 24), GSAGSAAGSGEF (SEQ ID NO: 25), and (XP)n, with X designating any amino acid, e.g., Ala, Lys, or Glu.
[0077] In various embodiments, the linker may be functional. For example, without limitation, the linker may function to improve the folding and/or stability, improve the expression, improve the
pharmacokinetics, and/or improve the bioactivity of the present compositions. In another example, the linker may function to target the compositions to a particular cell type or location.
[0078] In some embodiments, a gp96 peptide can be fused to the hinge, CH2 and CH3 domains of murine lgG1 (Bowen et al., J Immunol 1996, 156:442-449). This region of the lgG1 molecule contains three cysteine residues that normally are involved in disulfide bonding with other cysteines in the Ig molecule. Since none of the cysteines are required for the peptide to function as a tag, one or more of these cysteine residues can be substituted by another amino acid residue, such as, for example, serine.
[0079] Various leader sequences known in the art also can be used for efficient secretion of gp96-lg fusion proteins from bacterial and mammalian cells (see, von Heijne, J Mol Biol 1985, 184:99-105). Leader peptides can be selected based on the intended host cell, and may include bacterial, yeast, viral, animal, and mammalian sequences. For example, the herpes virus glycoprotein D leader peptide is suitable for use in a variety of mammalian cells. Another leader peptide for use in mammalian cells can be obtained from the V-J2-C region of the mouse immunoglobulin kappa chain (Bernard et al., Proc Natl Acad Sci USA 1981 , 78:5812-5816). DNA sequences encoding peptide tags or leader peptides are known or readily available from libraries or commercial suppliers, and are suitable in the fusion proteins described herein.
[0080] Furthermore, in various embodiments, one may substitute the gp96 of the present disclosure with one or more vaccine proteins. For instance, various heat shock proteins are among the vaccine proteins. In various embodiments, the heat shock protein is one or more of a small hsp, hsp40, hsp60, hsp70, hsp90, and hsp110 family member, inclusive of fragments, variants, mutants, derivatives or combinations thereof (Hickey, et al., 1989, Mol. Cell. Biol. 9:2615-2626; Jindal, 1989, Mol. Cell. Biol. 9:2279-2283).
T-Cell Co-Stimulation
[0081] The cell based therapy using the expression vectors provided herein can encode one or more biological response modifiers. In various embodiments, the cell based therapies can encode one or more T cell costimultory molecules.
[0082] In various embodiments, the cell based therapies allow for a robust, antigen-specific CD8 cytotoxic T lymphocyte (CTL) expansion. In various embodiments, the cell based therapies selectively enhance CD8 cytotoxic T lymphocyte (CTL) and do not substantially enhance T cell types that can be pro-tumor, and which include, but are not limited to, Tregs, CD4+ and/or CD8+ T cells expressing one or more checkpoint inhibitory receptors, Th2 cells and Th17 cells. Checkpoint inhibitory receptors refers to receptors (e.g., CTLA-4, B7-H3, B7-H4, TIM-3) expressed on immune cells that prevent or inhibit
uncontrolled immune responses. For instance, the present cell based therapies do not substantially enhance FOXP3+ regulatory T cells. In some embodiments, this selective CD8 T cell enhancement is in contrast to the non-specific T cell enhancement observed with a combination therapy of a gp-96 fusion and an antibody against a T cell costimultory molecule.
[0083] For example, the cell based therapies comprise an agonist of 0X40 {e.g., an 0X40 ligand-lg (OX40L-lg) fusion, or a fragment thereof that binds 0X40), an agonist of inducible T-cell costimulator (ICOS) {e.g., an ICOS ligand-lg (ICOSL-lg) fusion, or a fragment thereof that binds ICOS), an agonist of CD40 (e.g., a CD40L-lg fusion protein, or fragment thereof), an agonist of CD27 (e.g., a CD70-lg fusion protein or fragment thereof), or an agonist of 4-1 BB (e.g., a 4-1 BB ligand-lg (4-1 BBL-lg) fusion, or a fragment thereof that binds 4-1 BB). In some embodiments, the cell based therapies comprise a vector which encodes an agonist of TNFRSF25 (e.g., a TL1 A-lg fusion, or a fragment thereof that binds TNFRSF25), or an agonist of glucocorticoid-induced tumor necrosis factor receptor (GITR) (e.g., a GITR ligand-lg (GITRL-lg) fusion, or a fragment thereof that binds GITR), or an agonist of CD40 (e.g., a CD40 ligand-lg (CD40L-lg) fusion, or a fragment thereof that binds CD40); or an agonist of CD27 (e.g., a CD27 ligand-lg (e.g. CD70L-lg) fusion, or a fragment thereof that binds CD40).
[0084] ICOS is an inducible T cell costimulatory receptor molecule that displays some homology to CD28 and CTLA-4, and interacts with B7-H2 expressed on the surface of antigen-presenting cells. ICOS has been implicated in the regulation of cell-mediated and humoral immune responses.
[0085] 4-1 BB is a type 2 transmembrane glycoprotein belonging to the TNF superfamily, and is expressed on activated T Lymphocytes.
[0086] 0X40 (also referred to as CD134 or TNFRSF4) is a T cell costimulatory molecule that is engaged by OX40L, and frequently is induced in antigen presenting cells and other cell types. 0X40 is known to enhance cytokine expression and survival of effector T cells.
[0087] GITR (TNFRSF18) is a T cell costimulatory molecule that is engaged by GITRL and is preferentially expressed in FoxP3+ regulatory T cells. GITR plays a significant role in the maintenance and function of Treg within the tumor microenvironment.
[0088] TNFRSF25 is a T cell costimulatory molecule that is preferentially expressed in CD4+ and CD8+ T cells following antigen stimulation. Signaling through TNFRSF25 is provided by TL1A, and functions to enhance T cell sensitivity to IL-2 receptor mediated proliferation in a cognate antigen dependent manner.
[0089] CD40 is a costimulatory protein found on various antigen presenting cells which plays a role in their activation. The binding of CD40L (CD154) on TH cells to CD40 activates antigen presenting cells and induces a variety of downstream effects.
[0090] CD27 a T cell costimulatory molecule belonging to the TNF superfamily which plays a role in the
generation and long-term maintenance of T cell immunity. It binds to a ligand CD70 in various immunological processes.
[0091] Additional costimulatory molecules that may be utilized in the present invention include, but are not limited to, HVEM, CD28, CD30, CD30L, CD40, CD70, LIGHT (CD258), B7-1 , and B7-2.
[0092] As for the gp96-lg fusions, the Ig portion ("tag”) of the T cell costimulatory fusion protein can contain, a non-variable portion of an immunoglobulin molecule {e.g., an lgG1 , lgG2, lgG3, lgG4, IgM, IgA, or IgE molecule). As described above, such portions typically contain at least functional CH2 and CH3 domains of the constant region of an immunoglobulin heavy chain. In some embodiments, a T cell costimulatory peptide can be fused to the hinge, CH2 and CH3 domains of murine lgG1 (Bowen et al., J Immunol 1996, 156:442-449). The Ig tag can be from a mammalian {e.g., human, mouse, monkey, or rat) immunoglobulin, but human immunoglobulin can be particularly useful when the fusion protein is intended for in vivo use for humans. Again, DNAs encoding immunoglobulin light or heavy chain constant regions are known or readily available from cDNA libraries. Various leader sequences as described above also can be used for secretion of T cell costimulatory fusion proteins from bacterial and mammalian cells.
[0093] A representative nucleotide sequence (SEQ ID NO:4) encoding the extracellular domain of human ICOSL fused to Ig, and the amino acid sequence of the encoded fusion (SEQ ID NO:5) are provided:
AT GAGACT GGGAAGCCCT GGCCT GCT GTTT CT GCT GTT CAGCAGCCT GAGAGCCGACACCCAGG AAAAAGAAGT GCGGGCCAT GGT GGGAAGCGACGT GGAACT GAGCT GCGCCT GT CCT GAGGGCAG CAGATT CGACCT GAACGACGT GTACGT GTACT GGCAGACCAGCGAGAGCAAGACCGT CGT GACCT ACCACAT CCCCCAG AACAGCT CCCT GG AAAACGT GGACAGCCGGT ACAGAAACCGGGCCCT GAT GT CT CCT GCCGGCAT GCT GAGAGGCGACTT CAGCCT GCGGCT GTT CAACGT GACCCCCCAGGAC GAGCAGAAATT CCACT GCCT GGT GCT GAGCCAGAGCCT GGGCTT CCAGGAAGT GCT GAGCGT GG AAGT GACCCT GCACGT GGCCGCCAATTT CAGCGT GCCAGT GGT GT CT GCCCCCCACAGCCCTT CT CAGGAT GAGCT GACCTT CACCT GT ACCAGCAT CAACGGCT ACCCCAGACCCAAT GT GT ACT GGAT CAACAAGACCGACAACAGCCT GCT GGACCAGGCCCT GCAGAACGATACCGT GTT CCT GAACAT GC GGGGCCT GTACGACGT GGT GT CCGT GCT GAGAAT CGCCAGAACCCCCAGCGT GAACAT CGGCT G CT GCAT CGAGAACGT GCT GCT GCAGCAGAACCT GACCGT GGGCAGCCAGACCGGCAACGACAT C GGCGAGAGAGACAAGAT CACCGAGAACCCCGT GT CCACCGGCGAGAAGAAT GCCGCCACCT CT A AGT ACGGCCCT CCCT GCCCTT CTT GCCCAGCCCCT GAATTT CT GGGCGGACCCT CCGT GTTT CT G TT CCCCCCAAAGCCCAAGGACACCCT GAT GAT CAGCCGGACCCCCGAAGT GACCT GCGT GGT GG T GGAT GT GT CCCAGGAAGAT CCCGAGGT GCAGTT CAATT GGTACGT GGACGGGGT GGAAGT GCA CAACGCCAAGACCAAGCCCAGAGAGGAACAGTT CAACAGCACCT ACCGGGT GGT GT CT GT GCT G ACCGT GCT GCACCAGGATT GGCT GAGCGGCAAAGAGT ACAAGT GCAAGGT GT CCAGCAAGGGCC T GCCCAGCAGCAT CGAAAAGACCAT CAGCAACGCCACCGGCCAGCCCAGGGAACCCCAGGT GTA CACACT GCCCCCT AGCCAGGAAGAGAT GACCAAGAACCAGGT GT CCCT GACCT GT CT CGT GAAGG GCTT CT ACCCCT CCGAT AT CGCCGT GGAAT GGGAGAGCAACGGCCAGCCAGAGAACAACT ACAAG ACCACCCCCCCAGT GCT GGACAGCGACGGCT CATT CTT CCT GTACT CCCGGCT GACAGT GGACAA GAGCAGCT GGCAGGAAGGCAACGT GTT CAGCT GCAGCGT GAT GCACGAAGCCCT GCACAACCAC TACACCCAGAAGT CCCT GT CT CT GT CCCT GGGCAAAT GA (SEQ ID NO:4).
MRLGSPGLLFLLFSSLRADTQEKEVRAMVGSDVELSCACPEGSRFDLNDVYVYWQTSESKTVVTYHIP
QNSSLENVDSRYRNRALMSPAGMLRGDFSLRLFNVTPQDEQKFHCLVLSQSLGFQEVLSVEVTLHVA
ANFSVPVVSAPHSPSQDELTFTCTSINGYPRPNVYWINKTDNSLLDQALQNDTVFLNMRGLYDVVSVL
RIARTPSVNIGCCIENVLLQQNLTVGSQTGNDIGERDKITENPVSTGEKNAATSKYGPPCPSCPAPEFLG
GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVV
SVLTVLHQDWLSGKEYKCKVSSKGLPSSIEKTISNATGQPREPQVYTLPPSQEEMTKNQVSLTCLVKG
FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSSWQEGNVFSCSVMHEALHNHYTQ
KSLSLSLGK (SEQ ID NO:5).
[0094] A representative nucleotide sequence (SEQ ID NO:6) encoding the extracellular domain of human 4-1 BBL fused to Ig, and the encoded amino acid sequence (SEQ ID NO:7) are provided:
AT GT CT AAGT ACGGCCCTCCCTGCCCTAGCT GCCCT GCCCCT GAATTT CT GGGCGGACCCAGCGT GTT CCT GTT CCCCCCAAAGCCCAAGGACACCCT GAT GAT CAGCCGGACCCCCGAAGT GACCT GC GT GGT GGT GGAT GT GT CCCAGGAAGAT CCCGAGGT GCAGTT CAATT GGTACGT GGACGGCGT GG AAGT GCACAACGCCAAGACCAAGCCCAG AGAGGAACAGTT CAACAGCACCT ACCGGGTGGTGTC CGT GCT GACCGT GCT GCACCAGGATT GGCT GAGCGGCAAAGAGT ACAAGT GCAAGGT GT CCAGC AAGGGCCT GCCCAGCAGCAT CGAGAAAACCAT CAGCAACGCCACCGGCCAGCCCAGGGAACCCC AGGT GTACACACT GCCCCCTAGCCAGGAAGAGAT GACCAAGAACCAGGT GT CCCT GACCT GT CT C GT GAAGGGCTT CT ACCCCT CCGAT AT CGCCGT GGAAT GGGAGAGCAACGGCCAGCCT GAGAACA ACT ACAAGACCACCCCCCCAGT GCT GGACAGCGACGGCT CATT CTT CCT GT ACAGCAGACT GACC GT GGACAAGAGCAGCT GGCAGGAAGGCAACGT GTT CAGCT GCAGCGT GAT GCACGAGGCCCT GC ACAACCACTACACCCAGAAGT CCCT GT CT CT GAGCCT GGGCAAGGCCT GT CCAT GGGCT GT GT CT GGCGCT AGAGCCT CT CCT GGAT CT GCCGCCAGCCCCAGACT GAGAGAGGGACCT GAGCT GAGCC CCGAT GAT CCT GCCGGACT GCT GGAT CT GAGACAGGGCAT GTT CGCCCAGCT GGT GGCCCAGAA CGT GCT GCT GAT CGAT GGCCCCCT GAGCT GGT ACAGCGAT CCT GGACT GGCT GGCGT GT CACT G ACAGGCGGCCT GAGCTACAAAGAGGACACCAAAGAACT GGT GGT GGCCAAGGCCGGCGT GT ACT ACGT GTT CTTT CAGCT GGAACT GCGGAGAGT GGT GGCCGGCGAAGGAT CCGGCT CT GT GT CT CT GGCT CT GCAT CT GCAGCCCCT GAGAT CT GCT GCT GGCGCT GCT GCT CT GGCCCT GACAGT GGAC CT GCCT CCT GCCT CT AGCGAGGCCAGAAACAGCGCATT CGGGTTT CAAGGCAGACT GCT GCACCT GT CT GCCGGCCAGAGACT GGGAGT GCAT CT GCACACAGAGGCCAGAGCCAGGCACGCCT GGCA GCT GACT CAGGGCGCTACAGT GCT GGGCCT GTT CAGAGT GACCCCCGAGATT CCAGCCGGCCT G CCT AGCCCCAGAT CCGAAT GA (SEQ ID NO:6)
MSKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVH
NAKTKPREEQFNSTYRVVSVLTVLHQDWLSGKEYKCKVSSKGLPSSIEKTISNATGQPREPQVYTLPPS
QEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSSWQEGN
VFSCSVMHEALHNHYTQKSLSLSLGKACPWAVSGARASPGSAASPRLREGPELSPDDPAGLLDLRQG
MFAQLVAQNVLLIDGPLSWYSDPGLAGVSLTGGLSYKEDTKELWAKAGVYYVFFQLELRRVVAGEGS
GSVSLALHLQPLRSAAGAAALALTVDLPPASSEARNSAFGFQGRLLHLSAGQRLGVHLHTEARARHAW
QLTQGATVLGLFRVTPEIPAGLPSPRSE (SEQ ID NO:7).
[0095] A representative nucleotide sequence (SEQ ID NO:8) encoding the extracellular domain of human TL1 A fused to Ig, and the encoded amino acid sequence (SEQ ID NO:9) are provided:
AT GT CT AAGT ACGGCCCTCCCTGCCCTAGCT GCCCT GCCCCT GAATTT CT GGGCGGACCCAGCGT GTT CCT GTT CCCCCCAAAGCCCAAGGACACCCT GAT GAT CAGCCGGACCCCCGAAGT GACCT GC GT GGT GGT GGAT GT GT CCCAGGAAGAT CCCGAGGT GCAGTT CAATT GGTACGT GGACGGCGT GG AAGT GCACAACGCCAAGACCAAGCCCAG AGAGGAACAGTT CAACAGCACCT ACCGGGTGGTGTC
CGT GCT GACCGT GCT GCACCAGGATT GGCT GAGCGGCAAAGAGT ACAAGT GCAAGGT GT CCAGC AAGGGCCT GCCCAGCAGCAT CGAGAAAACCAT CAGCAACGCCACCGGCCAGCCCAGGGAACCCC AGGT GTACACACT GCCCCCTAGCCAGGAAGAGAT GACCAAGAACCAGGT GT CCCT GACCT GT CT C GT GAAGGGCTT CT ACCCCT CCGAT AT CGCCGT GGAAT GGGAGAGCAACGGCCAGCCT GAGAACA ACT ACAAGACCACCCCCCCAGT GCT GGACAGCGACGGCT CATT CTT CCT GT ACAGCAGACT GACC GT GGACAAGAGCAGCT GGCAGGAAGGCAACGT GTT CAGCT GCAGCGT GAT GCACGAGGCCCT GC ACAACCACTACACCCAGAAGT CCCT GT CT CT GAGCCT GGGCAAGAT CGAGGGCCGGAT GGAT AGA GCCCAGGGCGAAGCCT GCGT GCAGTT CCAGGCT CT GAAGGGCCAGGAATT CGCCCCCAGCCACC AGCAGGT GT ACGCCCCT CT GAGAGCCGACGGCGAT AAGCCT AGAGCCCACCT GACAGT CGT GCG GCAGACCCCTACCCAGCACTT CAAGAAT CAGTT CCCCGCCCT GCACT GGGAGCACGAACT GGGC CT GGCCTT CACCAAGAACAGAAT GAACT ACACCAACAAGTTT CT GCT GAT CCCCGAGAGCGGCGA CT ACTT CAT CTACAGCCAAGT GACCTT CCGGGGCAT GACCAGCGAGT GCAGCGAGAT CAGACAGG CCGGCAGACCT AACAAGCCCG ACAGCAT CACCGT CGT GAT CACCAAAGT GACCGACAGCT ACCCC GAGCCCACCCAGCT GCT GAT GGGCACCAAGAGCGT GT GCGAAGT GGGCAGCAACT GGTT CCAGC CCAT CT ACCT GGGCGCCAT GTTT AGT CT GCAAGAGGGCGACAAGCT GAT GGT CAACGT GT CCGAC AT CAGCCT GGT GGATTACACCAAAGAGGACAAGACCTT CTT CGGCGCCTTT CT GCT CT GA (SEQ ID N0:8)
MSKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVH
NAKTKPREEQFNSTYRVVSVLTVLHQDWLSGKEYKCKVSSKGLPSSIEKTISNATGQPREPQVYTLPPS
QEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSSWQEGN
VFSCSVMHEALHNHYTQKSLSLSLGKIEGRMDRAQGEACVQFQALKGQEFAPSHQQVYAPLRADGDK
PRAHLTWRQTPTQHFKNQFPALHWEHELGLAFTKNRMNYTNKFLLIPESGDYFIYSQVTFRGMTSEC
SEIRQAGRPNKPDSITWITKVTDSYPEPTQLLMGTKSVCEVGSNWFQPIYLGAMFSLQEGDKLMVNVS
DISLVDYTKEDKTFFGAFLL (SEQ ID N0:9).
[0096] A representative nucleotide sequence (SEQ ID NO:10) encoding human OX40L-lg, and the encoded amino acid sequence (SEQ ID NO:11) are provided:
AT GT CT AAGT ACGGCCCTCCCTGCCCTAGCT GCCCT GCCCCT GAATTT CT GGGCGGACCCAGCGT GTT CCT GTT CCCCCCAAAGCCCAAGGACACCCT GAT GAT CAGCCGGACCCCCGAAGT GACCT GC GT GGT GGT GGAT GT GT CCCAGGAAGAT CCCGAGGT GCAGTT CAATT GGTACGT GGACGGCGT GG AAGT GCACAACGCCAAGACCAAGCCCAG AGAGGAACAGTT CAACAGCACCT ACCGGGTGGTGTC CGT GCT GACCGT GCT GCACCAGGATT GGCT GAGCGGCAAAGAGT ACAAGT GCAAGGT GT CCAGC AAGGGCCT GCCCAGCAGCAT CGAGAAAACCAT CAGCAACGCCACCGGCCAGCCCAGGGAACCCC AGGT GTACACACT GCCCCCTAGCCAGGAAGAGAT GACCAAGAACCAGGT GT CCCT GACCT GT CT C GT GAAGGGCTT CT ACCCCT CCGAT AT CGCCGT GGAAT GGGAGAGCAACGGCCAGCCT GAGAACA ACT ACAAGACCACCCCCCCAGT GCT GGACAGCGACGGCT CATT CTT CCT GT ACAGCAGACT GACC GT GGACAAGAGCAGCT GGCAGGAAGGCAACGT GTT CAGCT GCAGCGT GAT GCACGAGGCCCT GC ACAACCACT ACACCCAGAAGT CCCT GT CT CT GAGCCT GGGCAAGAT CGAGGGCCGGAT GGAT CA GGT GT CACACAGAT ACCCCCGGAT CCAGAGCAT CAAAGT GCAGTTT ACCGAGTACAAG AAAG AGA AGGGCTTT AT CCT GACCAGCCAGAAAGAGGACGAGAT CAT GAAGGT GCAGAACAACAGCGT GAT C AT CAACT GCGACGGGTT CT ACCT GAT CAGCCT GAAGGGCT ACTT CAGT CAGGAAGT GAACAT CAG CCT GCACTACCAGAAGGACGAGGAACCCCT GTT CCAGCT GAAGAAAGT GCGGAGCGT GAACAGC CT GAT GGTGGCCTCTCT GACCT ACAAGGACAAGGT GT ACCT G AACGT GACCACCGACAACACCAG CCT GGACGACTT CCACGT GAACGGCGGCGAGCT GAT CCT GATT CACCAGAACCCCGGCGAGTT C TGCGTGCTCTGA (SEQ ID NO:10)
MSKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVH
NAKTKPREEQFNSTYRVVSVLTVLHQDWLSGKEYKCKVSSKGLPSSIEKTISNATGQPREPQVYTLPPS
QEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSSWQEGN VFSCSVMHEALHNHYTQKSLSLSLGKIEGRMDQVSHRYPRIQSIKVQFTEYKKEKGFILTSQKEDEIMKV QNNSVIINCDGFYLISLKGYFSQEVNISLHYQKDEEPLFQLKKVRSVNSLMVASLTYKDKVYLNVTTDNT SLDDFHVNGGELILIHQNPGEFCVL (SEQ ID NO:11).
[0097] Representative nucleotide and amino acid sequences for human TL1A are set forth in SEQ ID NO:12 and SEQ ID NO:13, respectively:
T CCCAAGT AGCT GGGACT ACAGGAGCCCACCACCACCCCCGGCT AATTTTTT GT ATTTTT AGT AGA GACGGGGTTT CACCGT GTT AGCCAAGAT GGT CTT GAT CACCT GACCT CGT GAT CCACCCGCCTT G GCCT CCCAAAGT GCT GGGATTACAGGCAT GAGCCACCGCGCCCGGCCT CCATT CAAGT CTTTATT GAATAT CT GCTAT GTT CTACACACT GTT CTAGGT GCT GGGGAT GCAACAGGGGACAAAATAGGCAA AAT CCCTGT CCTTTT GGGGTT GACATT CT AGT GACT CTT CAT GT AGT CT AGAAGAAGCT CAGT GAAT AGT GT CT GT GGTT GTT ACCAGGGACACAAT GACAGGAACATT CTT GGGTAGAGT GAGAGGCCT GG GGAGGGAAGGGT CT CTAGGAT GGAGCAGAT GCT GGGCAGT CTTAGGGAGCCCCT CCT GGCAT GC ACCCCCT CAT CCCT CAGGCCACCCCCGT CCCTT GCAGGAGCACCCT GGGGAGCT GT CCAGAGCG CT GT GCCGCT GT CT GT GGCT GGAGGCAGAGTAGGT GGT GT GCT GGGAAT GCGAGT GGGAGAACT GGGAT GGACCGAGGGGAGGCGGGT GAGGAGGGGGGCAACCACCCAACACCCACCAGCT GCTTT CAGT GTT CT GGGT CCAGGT GCT CCT GGCT GGCCTT GT GGT CCCCCT CCT GCTT GGGGCCACCCT GACCT ACACAT ACCGCCACT GCTGGCCT CACAAGCCCCT GGTT ACT GCAGAT GAAGCT GGGAT GG AGGCT CT GACCCCACCACCGGCCACCCAT CT GT CACCCTT GGACAGCGCCCACACCCTT CT AGCA CCT CCT GACAGCAGT GAGAAGAT CT GCACCGT CCAGTT GGT GGGTAACAGCT GGACCCCT GGCTA CCCCGAGACCCAGGAGGCGCT CT GCCCGCAGGT GACAT GGT CCT GGGACCAGTT GCCCAGCAGA GCT CTT GGCCCCGCT GCT GCGCCCACACT CT CGCCAGAGT CCCCAGCCGGCT CGCCAGCCAT GA T GCT GCAGCCGGGCCCGCAGCT CT ACGACGT GAT GGACGCGGT CCCAGCGCGGCGCT GGAAGG AGTT CGT GCGCACGCT GGGGCT GCGCGAGGCAGAGAT CGAAGCCGT GGAGGT GGAGAT CGGCC GCTT CCGAGACCAGCAGTACGAGAT GCT CAAGCGCT GGCGCCAGCAGCAGCCCGCGGGCCT CG GAGCCGTTTACGCGGCCCT GGAGCGCAT GGGGCT GGACGGCT GCGT GGAAGACTT GCGCAGCC GCCT GCAGCGCGGCCCGT GACACGGCGCCCACTT GCCACCT AGGCGCT CT GGT GGCCCTT GCA GAAGCCCT AAGT ACGGTT ACTT ATGCGT GT AGACATTTT AT GT CACTT ATT AAGCCGCT GGCACGG CCCT GCGT AGCAGCACCAGCCGGCCCCACCCCT GCT CGCCCCT AT CGCT CCAGCCAAGGCGAAG AAGCACGAACGAAT GT CGAGAGGGGGT GAAGACATTT CT CAACTT CT CGGCCGGAGTTT GGCT GA GAT CGCGGTATTAAAT CT GT GAAAGAAAACAAAACAAAACAA (SEQ ID N0:12)
MEQRPRGCAAVAAALLLVLLGARAQGGTRSPRCDCAGDFHKKIGLFCCRGCPAGHYLKAPCTEPCGN
STCLVCPQDTFLAWENHHNSECARCQACDEQASQVALENCSAVADTRCGCKPGWFVECQVSQCVS
SSPFYCQPCLDCGALHRHTRLLCSRRDTDCGTCLPGFYEHGDGCVSCPTPPPSLAGAPWGAVQSAV
PLSVAGGRVGVFWVQVLLAGLVVPLLLGATLTYTYRHCWPHKPLVTADEAGMEALTPPPATHLSPLDS
AHTLLAPPDSSEKICTVQLVGNSWTPGYPETQEALCPQVTWSWDQLPSRALGPAAAPTLSPESPAGS
PAMMLQPGPQLYDVMDAVPARRWKEFVRTLGLREAEIEAVEVEIGRFRDQQYEMLKRWRQQQPAGL
GAVYAALERMGLDGCVEDLRSRLQRGP (SEQ ID N0:13).
[0098] Representative nucleotide and amino acid sequences for human HVEM are set forth in SEQ ID
NO:26 (accession no. CR456909) and SEQ ID NO:27, respectively (accession no. CR456909):
AT GGAGCCT CCT GGAGACT GGGGGCCT CCT CCCT GGAGAT CCACCCCCAAAACCGACGT CTT GA GGCT GGT GCT GT AT CT CACCTT CCT GGGAGCCCCCT GCT ACGCCCCAGCT CT GCCGT CCT GCAAG GAGGACGAGT ACCCAGT GGGCT CCGAGT GCT GCCCCAAGT GCAGT CCAGGTT AT CGT GT GAAGG AGGCCT GCGGGGAGCT GACGGGCACAGT GT GT GAACCCT GCCCT CCAGGCACCT ACATT GCCCA CCT CAAT GGCCTAAGCAAGT GT CT GCAGT GCCAAAT GT GT GACCCAGCCAT GGGCCT GCGCGCG
AGCCGGAACT GCT CCAGGACAGAGAACGCCGT GT GT GGCT GCAGCCCAGGCCACTT CT GCAT CG T CCAGGACGGGGACCACT GCGCCGCGT GCCGCGCTT ACGCCACCT CCAGCCCGGGCCAGAGGG T GCAGAAGGGAGGCACCGAGAGT CAGGACACCCT GT GT CAGAACT GCCCCCCGGGGACCTT CT C T CCCAAT GGGACCCT GGAGGAAT GT CAGCACCAGACCAAGT GCAGCT GGCT GGT GACGAAGGCC GGAGCT GGGACCAGCAGCT CCCACT GGGTAT GGT GGTTT CT CT CAGGGAGCCT CGT CAT CGT CAT T GTTT GCT CCACAGTT GGCCT AAT CAT AT GT GT GAAAAGAAG AAAGCCAAGGGGT GAT GT AGT CAA GGT GAT CGT CT CCGT CCAGCGGAAAAGACAGGAGGCAGAAGGT GAGGCCACAGT CATT GAGGCC CT GCAGGCCCCT CCGGACGT CACCACGGT GGCCGT GGAGGAGACAATACCCT CATT CACGGGGA GGAGCCCAAACCATTAA (SEQ ID NO:26)
MEPPGDWGPPPWRSTPKTDVLRLVLYLTFLGAPCYAPALPSCKEDEYPVGSECCPKCSPGYRVKEAC GELTGTVCEPCPPGTYIAHLNGLSKCLQCQMCDPAMGLRASRNCSRTENAVCGCSPGHFCIVQDGDH CAACRAYATSSPGQRVQKGGTESQDTLCQNCPPGTFSPNGTLEECQHQTKCSWLVTKAGAGTSSSH WVWWFLSGSLVIVIVCSTVGLIICVKRRKPRGDVVKVIVSVQRKRQEAEGEATVIEALQAPPDVTTVAVE ETIPSFTGRSPNH (SEQ ID NO:27).
[0099] Representative nucleotide and amino acid sequences for human CD28 are set forth in SEQ ID NO:28 (accession no. NM_006139) and SEQ ID NO:29, respectively:
T AAAGT CAT CAAAACAACGTT AT AT CCT GT GT GAAAT GCT GCAGT CAGGAT GCCTT GT GGTTT GAG T GCCTT GAT CAT GT GCCCT AAGGGGAT GGT GGCGGT GGT GGT GGCCGT GGAT GACGGAGACT CT CAGGCCTT GGCAGGT GCGT CTTT CAGTT CCCCT CACACTT CGGGTT CCT CGGGGAGGAGGGGCT GGAACCCT AGCCCAT CGT CAGGACAAAGAT GCT CAGGCT GCT CTT GGCT CT CAACTT ATT CCCTT C AATT CAAGT AACAGGAAACAAGATTTT GGT GAAGCAGT CGCCCAT GCTT GT AGCGT ACG ACAAT GC GGT CAACCTT AGCT GCAAGT ATT CCT ACAAT CT CTT CT CAAGGGAGTT CCGGGCAT CCCTT CACAA AGGACT GGAT AGTGCTGT GGAAGT CT GT GTT GTATAT GGGAATT ACT CCCAGCAGCTT CAGGTTT A CT CAAAAACGGGGTT CAACT GT GAT GGGAAATT GGGCAAT GAAT CAGT GACATT CTACCT CCAGAA TTT GT AT GTT AACCAAACAGAT ATTT ACTT CT GCAAAATT GAAGTT AT GT AT CCT CCT CCTT ACCT AG ACAAT GAGAAG AGCAAT GGAACCATT AT CCAT GT G AAAGGGAAACACCTTT GT CCAAGT CCCCT AT TT CCCGGACCTT CTAAGCCCTTTT GGGT GCT GGT GGT GGTT GGT GGAGT CCT GGCTT GCT AT AGC TT GCT AGT AACAGT GGCCTTT ATT ATTTT CTGGGT GAGGAGTAAGAGGAGCAGGCT CCT GCACAGT GACTACAT GAACAT GACT CCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTAT GCCC CACCACGCGACTT CGCAGCCTAT CGCT CCT GACACGGACGCCTAT CCAGAAGCCAGCCGGCT GG CAGCCCCCAT CT GCT CAAT AT CACT GCT CT GGATAGGAAAT GACCGCCAT CT CCAGCCGGCCACC T CAGGCCCCT GTT GGGCCACCAAT GCCAATTTTT CT CGAGT GACTAGACCAAAT AT CAAGAT CATT TT GAG ACT CT GAAAT GAAGT AAAAGAGATTT CCT GT GACAGGCCAAGT CTT ACAGT GCCAT GGCCC ACATT CCAACTT ACCAT GT ACTT AGT GACTT GACT GAGAAGTT AGGGT AGAAAACAAAAAGGGAGT GG ATT CT GGGAGCCT CTT CCCTTT CT CACT CACCT GCACAT CT CAGT CAAGCAAAGT GTGGTATCC ACAGACATTTT AGTT GCAGAAGAAAGGCT AGGAAAT CATT CCTTTT GGTT AAAT GGGT GTTT AAT CT TTT GGTTAGT GGGTT AAACGGGGT AAGTTAGAGT AGGGGGAGGGAT AGGAAGACAT ATTT AAAAA CCATT AAAACACT GT CT CCCACT CAT GAAAT GAGCCACGT AGTT CCT ATTT AAT GCT GTTTT CCTTT AGTTT AGAAAT ACAT AGACATT GT CTTTT AT GAATT CT GAT CAT ATTT AGT CATTTT GACCAAAT GAG GG ATTT GGT CAAAT GAGGGATT CCCT CAAAGCAAT AT CAGGT AAACCAAGTT GCTTT CCT CACT CC CT GT CAT GAG ACTT CAGT GTT AAT GTT CACAAT AT ACTTT CGAAAGAAT AAAAT AGTT CT CCT ACAT G AAGAAAGAAT AT GT CAGGAAAT AAGGT CACTTT AT GT CAAAATT ATTT GAGT ACT AT GGG ACCT GGC GCAGT GGCT CAT GCTT GT AAT CCCAGCACTTT GGGAGGCCGAGGT GGGCAGAT CACTT GAGAT CA GG ACCAGCCT GGT CAAGAT GGT GAAACT CCGTCT GTACT AAAAAT ACAAAATTT AGCTT GGCCTGG T GGCAGGCACCT GT AAT CCCAGCT GCCCAAGAGGCT GAGGCAT GAGAAT CGCTT GAACCT GGCA GGCGGAGGTT GCAGT GAGCCGAGATAGT GCCACAGCT CT CCAGCCT GGGCGACAGAGT GAGACT
CCAT CT CAAACAACAACAACAACAACAACAACAACAACAAACCACAAAATT ATTT GAGT ACT GT G AA GG ATT ATTT GT CT AACAGTT CATT CCAAT CAGACCAGGT AGGAGCTTT CCT GTTT CAT AT GTTT CAG GGTT GCACAGTT GGT CT CTTT AAT GTCGGT GT GGAGAT CCAAAGT GGGTT GT GGAAAGAGCGT CC AT AGGAGAAGT GAGAAT ACT GT GAAAAAGGGAT GTT AGCATT CATT AGAGT AT GAGGAT GAGT CCC AAGAAGGTT CTTT GGAAGGAGGACGAAT AGAAT GGAGTAAT GAAATT CTT GCCAT GT GCT GAGGA GAT AGCCAGCATT AGGT GACAAT CTT CCAGAAGT GGT CAGGCAGAAGGT GCCCTGGT GAGAGCT C CTTT ACAGGGACTTTAT GT GGTTT AGGGCT CAGAGCT CCAAAACT CT GGGCT CAGCT GCT CCT GT A CCTT GGAGGT CCATT CACAT GGGAAAGTATTTT GGAAT GT GT CTTTT GAAGAGAGCAT CAGAGTT C TT AAGGGACT GGGT AAGGCCT GACCCT GAAAT GACCAT GGAT ATTTTT CT ACCT ACAGTTT GAGT C AACTAGAAT AT GCCT GGGGACCTT GAAGAAT GGCCCTT CAGT GGCCCT CACCATTT GTT CAT GCTT CAGTT AATT CAGGT GTT GAAGG AGCTT AGGTTTT AGAGGCACGT AGACTT GGTT CAAGT CT CGTT A GT AGTT GAAT AGCCT CAGGCAAGT CACT GCCCACCT AAGAT GAT GGTT CTT CAACT AT AAAAT GGA GATAATGGTTACAAAT GT CT CTT CCTATAGTATAAT CT CCATAAGGGCAT GGCCCAAGT CT GT CTTT GACT CTGCCTATCCCT GACATTT AGT AGCAT GCCCGACAT ACAAT GTT AGCT ATT GGT ATT ATT GCC ATATAGATAAATTATGTATAAAAATTAAACTGGGCAATAGCCTAAGAAGGGGGGAATATTGTAACAC AAATTT AAACCCACT ACGCAGGG AT GAGGT GCT AT AAT AT GAGGACCTTTT AACTT CCAT CATTTT C CT GTTT CTT GAAAT AGTTT AT CTT GT AAT GAAAT AT AAGGCACCT CCCACTTTT AT GT AT AGAAAGAG GT CTTTT AATTTTTTTTT AAT GT GAGAAGGAAGGGAGGAGT AGGAAT CTT GAG ATT CCAGAT CGAAA AT ACT GTACTTT GGTT GATTTTTAAGT GGGCTT CCATT CCAT GGATTTAAT CAGT CCCAAGAAGAT C AAACT CAGCAGTACTT GGGT GCT GAAGAACT GTT GGATTTACCCT GGCACGT GT GCCACTT GCCA GCTT CTT GGGCACACAGAGTT CTT CAAT CCAAGTT AT CAGATT GTATTT GAAAAT GACAGAGCT GG AGAGTTTTTT GAAAT GGCAGT GGCAAAT AAAT AAAT ACTTTTTTTT AAAT GGAAAGACTT GAT CT AT G GT AAT AAAT GATTTT GTTTT CT GACT GGAAAAAT AGGCCT ACT AAAGAT GAAT CACACTT GAGAT GT TT CTT ACT CACT CT GCACAGAAACAAAGAAGAAAT GTT AT ACAGGGAAGT CCGTTTT CACT ATT AGT AT GAACCAAGAAAT GGTT CAAAAACAGT GGTAGGAGCAAT GCTTT CATAGTTT CAGATAT GGTAGTT AT GAAGAAAACAAT GT CATTT GCTGCT ATT ATT GTAAGAGT CTT AT AATT AAT GGT ACT CCT AT AATT TTT GATT GT GAGCT CACCT ATTT GGGTT AAGCAT GCCAATTT AAAGAGACCAAGT GTAT GTACATT A T GTT CT ACAT ATT CAGT GAT AAAATT ACT AAACT ACT AT AT GT CT GCTTT AAATTT GT ACTTT AAT ATT GT CTTTT GGT ATT AAGAAAGAT AT GCTTT CAGAAT AGAT AT GCTT CGCTTT GGCAAGGAATTT GGAT AGAACTT GCT ATTT AAAAGAGGT GT GGGGTAAAT CCTT GT AT AAAT CT CCAGTTT AGCCTTTTTT G A AAAAGCT AGACTTT CAAAT ACT AATTT CACTT CAAGCAGGGTACGTTT CT GGTTT GTTT GCTT GACT T CAGT CACAATTT CTT AT CAGACCAAT GGCT GACCT CTTT GAGAT GT CAGGCT AGGCTT ACCT AT GT GTT CT GT GT CAT GT GAAT GCT GAGAAGTTT GACAGAGAT CCAACTT CAGCCTT GACCCCAT CAGT C CCT CGGGTT AACT AACT GAGCCACCGGT CCT CAT GGCT ATTTT AAT G AGGGTATT GAT GGTT AAAT GCAT GTCT GAT CCCTT AT CCCAGCCATTT GCACT GCCAGCT GGGAACT AT ACCAGACCT GGAT ACT GAT CCCAAAGT GTT AAATT CAACT ACAT GCT GGAGATT AGAG AT GGT GCCAAT AAAGGACCCAGAA CCAGGAT CTT GATT GCT AT AGACTT ATT AAT AAT CCAGGT CAAAG AGAGT GACACACACT CT CT CAA GACCT GGGGT GAGGGAGT CT GT GTTAT CT GCAAGGCCATTT GAGGCT CAGAAAGT CT CT CTTT CC T AT AGAT AT AT GCAT ACTTT CT G ACAT AT AGGAAT GT AT CAGGAAT ACT CAACCAT CACAGGCAT GT TCCT ACCT CAGGGCCTTT ACAT GT CCT GTTT ACT CT GT CT AGAAT GT CCTT CT GT AGAT GACCT GGC TT GCCT CGT CACCCTT CAGGT CCTT GCT CAAGT GT CAT CTT CT CCCCT AGTT AAACT ACCCCACAC CCT GT CT GCTTT CCTT GCTT ATTTTT CT CCAT AGCATTTT ACCAT CT CTT ACATT AGACATTTTT CTT A TTT ATTT GT AGTTT AT AAGCTT CAT GAGGCAAGT AACTTT GCTTT GTTT CTT GCT GT AT CT CCAGT GC CCAGAGCAGT GCCTGGTAT AT AAT AAAT ATTT ATT GACT GAGT GAAAAAAAAAAAAAAAAA (SEQ ID NO:28)
MLRLLLALNLFPSIQVTGNKILVKQSPMLVAYDNAVNLSCKYSYNLFSREFRASLHKGLDSAVEVCVVY
GNYSQQLQVYSKTGFNCDGKLGNESVTFYLQNLYVNQTDIYFCKIEVMYPPPYLDNEKSNGTIIHVKGK
HLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHY
QPYAPPRDFAAYRS (SEQ ID NO:29).
[00100] Representative nucleotide and amino acid sequences for human CD30L are set forth in SEQ ID NO:30 (accession no. L09753) and SEQ ID NO:31 , respectively:
CCAAGT CACAT GATT CAGGATT CAGGGGGAGAAT CCTT CTT GGAACAGAGAT GGGCCCAGAACT G AAT CAGAT GAAGAGAGAT AAGGT GT GAT GT GGGGAAGACT AT AT AAAGAAT GGACCCAGGGCT GC AGCAAGCACT CAACGGAAT GGCCCCT CCT GGAGACACAGCCAT GCAT GT GCCGGCGGGCT CCGT GGCCAGCCACCT GGGGACCACGAGCCGCAGCT ATTT CT ATTT GACCACAGCCACT CTGGCT CT GT GCCTT GT CTT CACGGT GGCCACT ATT AT GGT GTT GGT CGTT CAGAGGACGGACT CCATT CCCAACT CACCT GACAACGT CCCCCT CAAAGGAGGAAATT GCT CAGAAGACCT CTTAT GTAT CCT GAAAAGAG CT CCATT CAAGAAGT CAT GGGCCTACCT CCAAGT GGCAAAGCAT CTAAACAAAACCAAGTT GT CTT GG AACAAAGAT GGCATT CT CCAT GG AGT CAGAT AT CAGGAT GGGAAT CT GGT GAT CCAATT CCCTG GTTT GT ACTT CAT CATTT GCCAACT GCAGTTT CTT GT ACAAT GCCCAAAT AATT CT GT CGAT CT GAA GTT GGAGCTT CT CAT CAACAAGCATAT CAAAAAACAGGCCCT GGT GACAGT GT GT GAGT CT GGAAT GCAAACGAAACACGT AT ACCAGAAT CT CT CT CAATT CTT GCT GGATT ACCT GCAGGT CAACACCAC CAT AT CAGT CAAT GT GG AT ACATT CCAGT ACAT AGAT ACAAGCACCTTT CCT CTT GAG AAT GTGTTG T CCAT CTT CTT AT ACAGT AATT CAGACT GAACAGTTT CT CTT GGCCTT CAGGAAGAAAGCGCCT CT C T ACCAT ACAGT ATTT CAT CCCT CCAAACACTT GGGCAAAAAGAAAACTTT AGACCAAGACAAACT AC ACAGGGT ATT AAAT AGT AT ACTT CT CCTT CTGTCT CTT GGAAAGAT ACAGCT CCAGGGTT AAAAAGA GAGTTTTT AGT GAAGTAT CTTT CAGAT AGCAGGCAGGGAAGCAAT GTAGT GT GGT GGGCAG AGCC CCACACAGAAT CAGAAGGGAT GAAT GGAT GT CCCAGCCCAACCACTAATT CACT GTAT GGT CTT GA T CT ATTT CTT CT GTTTT G AGAGCCT CCAGTT AAAAT GGGGCTT CAGT ACCAGAGCAGCT AGCAACT CT GCCCT AAT GGGAAAT GAAGGGGAGCT GGGT GT GAGT GTTT ACACT GT GCCCTT CACGGGATAC TT CTTTT AT CT GCAGAT GGCCT AAT GCTT AGTT GT CCAAGT CGCGAT CAAGGACT CT CT CACACAG GAAACTT CCCT AT ACT GGCAGAT ACACTT GT GACT GAACCAT GCCCAGTTT ATGCCT GT CT GACT G T CACT CT GGCACT AGGAGGCT GAT CTT GTACT CCAT AT GACCCCACCCCT AGGAACCCCCAGGG A AAACCAGGCT CGGACAGCCCCCT GTT CCT GAGAT GGAAAGCACAAATTTAATACACCACCACAAT G GAAAACAAGTT CAAAGACTTTT ACTT ACAGAT CCT GGACAGAAAGGGCAT AAT GAGT CT GAAGGGC AGT CCT CCTT CT CCAGGTTACAT GAGGCAGGAATAAGAAGT CAGACAGAGACAGCAAGACAGTTA ACAACGT AGGT AAAGAAAT AGGGT GT GGT CACT CT CAATT CACT GGCAAAT GCCT GAAT GGT CT GT CT GAAGGAAGCAACAGAGAAGT GGGGAAT CCAGT CT GCT AGGCAGGAAAGAT GCCTCT AAGTT CT T GT CT CT GGCCAGAGGT GTGGTAT AGAACCAGAAACCCAT AT CAAGGGT GACT AAGCCCGGCTT C CGGT AT GAGAAATT AAACTT GT AT ACAAAAT GGTT GCCAAGGCAACAT AAAATT AT AAGAATT C
(SEQ ID NO:30)
MDPGLQQALNGMAPPGDTAMHVPAGSVASHLGTTSRSYFYLTTATLALCLVFTVATIMVLWQRTDSIP NSPDNVPLKGGNCSEDLLCILKRAPFKKSWAYLQVAKHLNKTKLSWNKDGILHGVRYQDGNLVIQFPG LYFIICQLQFLVQCPNNSVDLKLELLINKHIKKQALVTVCESGMQTKHVYQNLSQFLLDYLQVNTTISVNV DTFQYIDTSTFPLENVLSIFLYSNSD (SEQ ID NO:31).
[00101] Representative nucleotide and amino acid sequences for human CD40 are set forth in SEQ ID NO:32 (accession no. NM_001250) and SEQ ID NO:33, respectively:
TTT CCT GGGCGGGGCCAAGGCT GGGGCAGGGGAGT CAGCAGAGGCCT CGCT CGGGCGCCCAGT GGT CCT GCCGCCT GGT CT CACCT CGCT AT GGTT CGT CT GCCT CT GCAGT GCGT CCT CT GGGGCT G CTT GCT GACCGCT GT CCAT CCAGAACCACCCACT GCAT GCAGAGAAAAACAGT ACCT AAT AAACAG T CAGT GCT GTT CTTT GT GCCAGCCAGGACAGAAACT GGT GAGT GACT GCACAGAGTT CACT GAAA CGGAAT GCCTT CCTT GCGGT GAAAGCGAATT CCT AGACACCT GGAACAGAGAGACACACT GCCAC
CAGCACAAATACT GCGACCCCAACCT AGGGCTT CGGGT CCAGCAGAAGGGCACCT CAGAAACAG ACACCAT CT GCACCT GT GAAGAAGGCT GGCACT GT ACGAGT GAGGCCT GT GAGAGCT GT GT CCT G CACCGCT CAT GCT CGCCCGGCTTT GGGGT CAAGCAGATT GCT ACAGGGGTTT CT GAT ACCAT CT G CGAGCCCT GCCCAGT CGGCTT CTT CT CCAAT GT GT CAT CT GCTTT CGAAAAAT GT CACCCTT GGAC AAGCT GT GAGACCAAAGACCT GGTT GT GCAACAGGCAGGCACAAACAAGACT GAT GTT GT CT GT G GT CCCCAGGAT CGGCT GAGAGCCCT GGT GGT GAT CCCCAT CAT CTT CGGGAT CCT GTTT GCCAT C CT CTT GGT GCT GGT CTTT AT CAAAAAGGT GGCCAAGAAGCCAACCAATAAGGCCCCCCACCCCAA GCAGGAACCCCAGGAGAT CAATTTT CCCGACGAT CTT CCT GGCT CCAACACT GCT GCT CCAGT GC AGGAGACTTT ACAT GGAT GCCAACCGGT CACCCAGGAGGAT GGCAAAGAGAGT CGCAT CT CAGT G CAGGAGAGACAGT GAGGCT GCACCCACCCAGGAGT GT GGCCACGT GGGCAAACAGGCAGTT GG CCAGAGAGCCT GGT GCT GCT GCT GCT GT GGCGT GAGGGT GAGGGGCT GGCACT GACT GGGCAT A GCT CCCCGCTT CT GCCT GCACCCCT GCAGTTT GAGACAGGAGACCT GGCACT GGAT GCAGAAACA GTT CACCTT GAAGAACCT CT CACTT CACCCT GGAGCCCAT CCAGT CT CCCAACTT GT ATT AAAG AC AGAGGCAGAAGTTT GGTGGTGGTGGT GTT GGGGTAT GGTTT AGT AAT AT CCACCAGACCTT CCGA T CCAGCAGTTT GGT GCCCAGAGAGGCAT CAT GGT GGCTT CCCT GCGCCCAGGAAGCCAT AT ACAC AGAT GCCCATT GCAGCATT GTTT GT GAT AGT GAACAACT GGAAGCT GCTT AACT GT CCAT CAGCAG GAGACT GGCT AAAT AAAATT AGAAT AT ATTT AT ACAACAGAAT CT CAAAAACACT GTT GAGT AAGGA AAAAAAGGCAT GCTGCT GAAT GAT GGGTAT GGAACTTTTT AAAAAAGT ACAT GCTTTT AT GT AT GT A T ATT GCCTAT GGAT AT ATGTAT AAAT ACAAT AT GCAT CAT AT ATT GAT AT AACAAGGGTT CT GGAAG GGTACACAGAAAACCCACAGCT CGAAGAGT GGT GACGT CT GGGGT GGGGAAGAAGGGT CT GGGG
G (SEQ ID NO:32)
MVRLPLQCVLWGCLLTAVHPEPPTACREKQYLINSQCCSLCQPGQKLVSDCTEFTETECLPCGESEFL DTWNRETHCHQHKYCDPNLGLRVQQKGTSETDTICTCEEGWHCTSEACESCVLHRSCSPGFGVKQIA TGVSDTICEPCPVGFFSNVSSAFEKCHPWTSCETKDLVVQQAGTNKTDWCGPQDRLRALVVIPIIFGIL FAILLVLVFIKKVAKKPTNKAPHPKQEPQEINFPDDLPGSNTAAPVQETLHGCQPVTQEDGKESRISVQE RQ (SEQ ID NO:33).
[00102] Representative nucleotide and amino acid sequences for human CD70 are set forth in SEQ ID NO:34 (accession no. NM_001252) and SEQ ID NO:35, respectively:
CCAGAGAGGGGCAGGCT GGT CCCCT GACAGGTT GAAGCAAGT AGACGCCCAGGAGCCCCGGGA GGGGGCT GCAGTTT CCTT CCTT CCTT CT CGGCAGCGCT CCGCGCCCCCAT CGCCCCT CCT GCGC TAGCGGAGGT GAT CGCCGCGGCGAT GCCGGAGGAGGGTT CGGGCT GCT CGGT GCGGCGCAGGC CCT AT GGGT GCGT CCT GCGGGCT GCTTT GGT CCCATT GGT CGCGGGCTT GGT GAT CT GCCT CGT GGT GT GCAT CCAGCGCTT CGCACAGGCT CAGCAGCAGCT GCCGCT CGAGT CACTT GGGT GGGAC GT AGCT GAGCT GCAGCT GAAT CACACAGGACCT CAGCAGGACCCCAGGCT AT ACT GGCAGGGGG GCCCAGCACT GGGCCGCT CCTT CCT GCAT GGACCAGAGCT GGACAAGGGGCAGCTACGTAT CCA TCGT GAT GGCAT CT ACAT GGT ACACAT CCAGGT GACGCT GGCCAT CT GCTCCT CCACGACGGCCT CCAGGCACCACCCCACCACCCT GGCCGT GGGAAT CT GCT CT CCCGCCT CCCGT AGCAT CAGCCT GCT GCGT CT CAGCTT CCACCAAGGTT GT ACCATT GCCT CCCAGCGCCT GACGCCCCT GGCCCGA GGGGACACACT CT GCACCAACCT CACT GGGACACTTTT GCCTT CCCGAAACACT GAT GAGACCTT CTTT GGAGT GCAGT GGGTGCGCCCCT G ACCACT GCTGCT GATT AGGGTTTTTT AAATTTT ATTTT AT TTTATTTAAGTT CAAGAGAAAAAGT GT ACACACAGGGGCCACCCGGGGTT GGGGT GGGAGT GT GG TGGGGGGT AGT GGT GGCAGGACAAGAGAAGGCATT GAGCTTTTT CTTT CATTTT CCT ATT AAAAAA TACAAAAATCA (SEQ ID NO:34)
MPEEGSGCSVRRRPYGCVLRAALVPLVAGLVICLVVCIQRFAQAQQQLPLESLGWDVAELQLNHTGP
QQDPRLYWQGGPALGRSFLHGPELDKGQLRIHRDGIYMVHIQVTLAICSSTTASRHHPTTLAVGICSPA
SRSISLLRLSFHQGCTIASQRLTPLARGDTLCTNLTGTLLPSRNTDETFFGVQWVRP(SEQ ID NO: 35).
[00103] Representative nucleotide and amino acid sequences for human LIGHT are set forth in SEQ ID NO:36 (accession no. CR541854) and SEQ ID NO:37, respectively:
AT GGAGGAGAGT GT CGT ACGGCCCT CAGT GTTT GT GGT GGAT GGACAGACCGACAT CCCATT CAC GAGGCT GGGACGAAGCCACCGGAGACAGT CGT GCAGT GT GGCCCGGGT GGGT CT GGGT CT CTT GOT GTT GOT GAT GGGGGCCGGGCT GGCCGT CCAAGGCT GGTT CCT CCT GCAGCT GCACT GGCGT CTAGGAGAGAT GGT CACCCGCCT GCCT GACGGACCT GCAGGCT CCT GGGAGCAGCT GATACAAG AGCGAAGGT CT CACGAGGT CAACCCAGCAGCGCAT CT CACAGGGGCCAACT CCAGCTT GACCGG CAGCGGGGGGCCGCT GTTAT GGGAGACT CAGCT GGGCCT GGCCTT CCT GAGGGGCCT CAGCT AC CACGAT GGGGCCCTT GTGGT CACCAAAGCT GGCT ACT ACT ACAT CT ACT CCAAGGT GCAGCT GGG CGGT GT GGGCT GCCCGCT GGGCCT GGCCAGCACCAT CACCCACGGCCT CT ACAAGCGCACACCC CGCT ACCCCGAGGAGCT GGAGCT GTT GGT CAGCCAGCAGT CACCCT GCGGACGGGCCACCAGCA GCT CCCGGGT CT GGT GGGACAGCAGCTT CCT GGGT GGT GT GGT ACACCT GGAGGCT GGGGAGG AGGT GGT CGT CCGT GT GCT GGAT GAACGCCT GGTT CGACT GCGT GAT GGTACCCGGT CTTACTT C GGGGCTTT CAT GGT GT GA (SEQ ID NO:36)
MEESWRPSVFVVDGQTDIPFTRLGRSHRRQSCSVARVGLGLLLLLMGAGLAVQGWFLLQLHWRLGE MVTRLPDGPAGSWEQLIQERRSHEVNPAAHLTGANSSLTGSGGPLLWETQLGLAFLRGLSYHDGALV VTKAGYYYIYSKVQLGGVGCPLGLASTITHGLYKRTPRYPEELELLVSQQSPCGRATSSSRVWWDSSF LGGWHLEAGEEWVRVLDERLVRLRDGTRSYFGAFMV (SEQ ID NO:37).
[00104] In various embodiments, the present invention provides for variants comprising any of the sequences described herein, for instance, a sequence having at least about 60%, or at least about 61%, or at least about 62%, or at least about 63%, or at least about 64%, or at least about 65%, or at least about 66%, or at least about 67%, or at least about 68%, or at least about 69%, or at least about 70%, or at least about 71%, or at least about 72%, or at least about 73%, or at least about 74%, or at least about 75%, or at least about 76%, or at least about 77%, or at least about 78%, or at least about 79%, or at least about 80%, or at least about 81%, or at least about 82%, or at least about 83%, or at least about 84%, or at least about 85%, or at least about 86%, or at least about 87%, or at least about 88%, or at least about 89%, or at least about 90%, or at least about 91 %, or at least about 92%, or at least about 93%, or at least about 94%, or at least about 95%, or at least about 96%, or at least about 97%, or at least about 98%, or at least about 99%) sequence identity with any of the sequences disclosed herein (for example, SEQ ID NOS: 1-13 and 26-37).
[00105] In various embodiments, the present invention provides for an amino acid sequence having one or more amino acid mutations relative any of the protein sequences described herein. In some embodiments, the one or more amino acid mutations may be independently selected from conservative or non-conservative substitutions, insertions, deletions, and truncations as described herein.
Checkpoint Blockade / Blockage of Tumor Immunosuppression
[00106] Some human tumors can be eliminated by a patient's immune system. For example, administration of a monoclonal antibody targeted to an immune "checkpoint” molecule can lead to complete response and tumor remission. A mode of action of such antibodies is through inhibition of an immune regulatory molecule that the tumors have co-opted as protection from an anti-tumor immune response. By inhibiting these "checkpoint” molecules {e.g., with an antagonistic antibody), a patient's CD8+ T cells may be allowed to proliferate and destroy tumor cells. For example, administration of a monoclonal antibody targeted to by way of example, without limitation, CTLA-4 or PD-1 can lead to complete response and tumor remission. The mode of action of such antibodies is through inhibition of CTLA-4 or PD-1 that the tumors have co-opted as protection from an anti-tumor immune response. By inhibiting these "checkpoint” molecules {e.g., with an antagonistic antibody), a patient's CD8+ T cells may be allowed to proliferate and destroy tumor cells.
[00107] Thus, the cell based therapies provided herein can be used in combination with one or more blocking antibodies targeted to an immune "checkpoint” molecule. For instance, in some embodiments, the cell based therapies provided herein can be used in combination with one or more blocking antibodies targeted to a molecule such as CTLA-4 or PD-1. For example, the cell based therapies provided herein may be used in combination with an agent that blocks, reduces and/or inhibits PD-1 and PD-L1 or PD- L2 and/or the binding of PD-1 with PD-L1 or PD-L2 (by way of non-limiting example, one or more of nivolumab (ONO-4538/BMS-936558, MDX1106, OPDIVO, BRISTOL MYERS SQUIBB), pembrolizumab (KEYTRUDA, Merck), pidilizumab (CT-011 , CURE TECH), MK-3475 (MERCK), BMS 936559 (BRISTOL MYERS SQUIBB), MPDL3280A (ROCHE)). In an embodiment, the cell based therapies provided herein may be used in combination with an agent that blocks, reduces and/or inhibits the activity of CTLA-4 and/or the binding of CTLA-4 with one or more receptors (e.g. CD80, CD86, AP2M1 , SHP-2, and PPP2R5A). For instance, in some embodiments, the immune-modulating agent is an antibody such as, by way of non-limitation, ipilimumab (MDX-010, M DX- 101 , Yervoy, BMS) and/or tremelimumab (Pfizer). Blocking antibodies against these molecules can be obtained from, for example, Bristol Myers Squibb (New York, NY), Merck (Kenilworth, NJ), Medlmmune (Gaithersburg, MD), and Pfizer (New York, NY).
[00108] Further, the cell based therapies provided herein can be used in combination with one or more blocking antibodies targeted to an immune "checkpoint” molecule such as for example, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160 (also referred to as BY55), CGEN-15049, CHK 1 and CHK2 kinases, A2aR, CEACAM {e.g., CEACAM-1 , CEACAM-3 and/or CEACAM-5), GITR, GITRL, galectin-9, CD244, CD160, TIGIT, SIRPa, ICOS, CD172a, and TMIGD2 and various B-7 family ligands (including, but are not limited to, B7-1 , B7-2, B7-DC, B7-H1 , B7-H2, B7-H3, B7-H4, B7-H5, B7-H6 and B7-H7).
Cell based therapies
[00109] The present disclosure provides a cell based therapy comprising a first cell containing an expression vector comprising, a nucleotide sequence that encode a secretable vaccine protein {e.g., a gp96-lg fusion protein) and a second cell containing an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein, {e.g., OX40L-lg or a portion thereof that binds specifically to 0X40, ICOSL-lg or a portion thereof that binds specifically to ICOS, 4-1 BBL-lg, or a portion thereof that binds specifically to 4-1 BBR, CD40L-lg, or a portion thereof that binds specifically to CD40, CD70-lg, or a portion thereof that binds specifically to CD27, TL1 A-lg or a portion thereof that binds specifically to TNFRSF25, or GITRL-lg or a portion thereof that binds specifically to GITR). In addition, present disclosure provides methods for making the cell based therapies described herein, as well as methods for administering the cell based therapies. In general, the methods provided herein include administering to a patient an effective amount of a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein, wherein the patient is undergoing a treatment with a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject.
[00110] In some embodiments, the methods provided herein include administering to a patient an effective amount of a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein, wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject, and wherein the patient is undergoing a treatment with a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein.
[00111] In some embodiments, the methods provided herein include administering to the patient an effective amount of a first cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject.
[00112] In some embodiments, gp96-lg based vaccines can be generated to stimulate antigen specific immune responses against individual antigens expressed by simian immunodeficiency virus, human immunodeficiency virus, hepatitis C virus and malaria. Immune responses to these vaccines may be enhanced through a cell based therapy of a T cell costimulatory fusion protein and a cell based therapy of gp96-lg.
[00113] cDNA or DNA sequences encoding a vaccine protein fusion (e.g., a gp96-lg fusion) and a T cell costimulatory fusion protein can be obtained (and, if desired, modified) using conventional DNA cloning and mutagenesis methods, DNA amplification methods, and/or synthetic methods. In general, a sequence encoding a vaccine protein fusion protein {e.g., a gp96-lg fusion protein) and/or a T cell costimulatory fusion protein can be inserted into a cloning vector for genetic modification and replication purposes prior to expression. Each coding sequence can be operably linked to a regulatory element, such as a promoter, for purposes of expressing the encoded protein in suitable host cells in vitro and in vivo.
[00114] The cell based therapy can be administered to produce secreted vaccine proteins (e.g., gp96- Ig) and T cell costimulatory fusion proteins. Cells may be cultured in vitro or genetically engineered, for example. Host cells can be obtained from normal or affected subjects, including healthy humans, cancer patients, and patients with an infectious disease, private laboratory deposits, public culture collections such as the American Type Culture Collection, or from commercial suppliers. Cells that can be used for production and secretion of gp96-lg fusion proteins and T cell costimulatory fusion proteins in vivo include, without limitation, epithelial cells, endothelial cells, keratinocytes, fibroblasts, muscle cells, hepatocytes; blood cells such as T lymphocytes, B lymphocytes, monocytes, macrophages, neutrophils, eosinophils, megakaryocytes, or granulocytes, various stem or progenitor cells, such as hematopoietic stem or progenitor cells (e.g., as obtained from bone marrow), umbilical cord blood, peripheral blood, fetal liver, etc., and tumor cells (e.g., human tumor cells). The choice of cell type depends on the type of tumor or infectious disease being treated or prevented, and can be determined by one of skill in the art.
[00115] Different host cells have characteristic and specific mechanisms for post-translational processing and modification of proteins. A host cell may be chosen which modifies and processes the expressed gene products in a specific fashion similar to the way the recipient processes its heat shock proteins (hsps). For the purpose of producing large amounts of gp96-lg, it can be preferable that the type of host cell has been used for expression of heterologous genes, and is reasonably well characterized and developed for large-scale production processes. In some embodiments, the host cells are autologous to the patient to whom the present fusion or recombinant cells secreting the present fusion proteins are subsequently administered.
[00116] In some embodiments, the cell based therapy as provided herein can be introduced into an antigenic cell. As used herein, antigenic cells can include preneoplastic cells that are infected with a cancer-causing infectious agent, such as a virus, but that are not yet neoplastic, or antigenic cells that have been exposed to a mutagen or cancer-causing agent, such as a DNA-damaging agent or radiation, for example. Other cells that can be used are preneoplastic cells that are in transition from a normal to a
neoplastic form as characterized by morphology or physiological or biochemical function.
[00117] Typically, the cancer cells and preneoplastic cells used in the methods provided herein are of mammalian origin. Mammals contemplated include humans, companion animals {e.g., dogs and cats), livestock animals {e.g., sheep, cattle, goats, pigs and horses), laboratory animals (e.g., mice, rats and rabbits), and captive or free wild animals.
[00118] In some embodiments, cancer cells (e.g., human tumor cells) can be used in the methods described herein. The cancer cells provide antigenic peptides that become associated non-covalently with the expressed gp96-lg fusion proteins. Cell lines derived from a preneoplastic lesion, cancer tissue, or cancer cells also can be used, provided that the cells of the cell line have at least one or more antigenic determinant in common with the antigens on the target cancer cells. Cancer tissues, cancer cells, cells infected with a cancer-causing agent, other preneoplastic cells, and cell lines of human origin can be used. Cancer cells excised from the patient to whom ultimately the fusion proteins ultimately are to be administered can be particularly useful, although allogeneic cells also can be used. In some embodiments, a cancer cell can be from an established tumor cell line such as, without limitation, an established non-small cell lung carcinoma (NSCLC), bladder cancer, melanoma, ovarian cancer, renal cell carcinoma, prostate carcinoma, sarcoma, breast carcinoma, squamous cell carcinoma, head and neck carcinoma, hepatocellular carcinoma, pancreatic carcinoma, or colon carcinoma cell line.
[00119] In various embodiments, the present fusion proteins allow for both the costimulation T cell and the presentation of various tumor cell antigens. For instance, in some embodiments, the present vaccine protein fusions (e.g., gp96 fusions) chaperone these various tumor antigens. In various embodiments, the tumor cells secrete a variety of antigens. Illustrative, but non-limiting, antigens that can be secreted are: ACRBP, ACTL8, ADAM2, ADAM29, AKAP3, AKAP4, ANKRD45, ARMC3, ARX, ATAD2, BAGE, BAGE2, BAGE3, BAGE4, BAGE5, BRDT, C15ORF60, C210RF99, CABYR, CAGE1 , CALR3, CASC5, CCDC110, CCDC33, CCDC36, CCDC62, CCDC83, CDCA1 , CEP290, CEP55, COX6B2, CPXCR1 , CRISP2, CSAG1 , CSAG2, CSAG3B, CT16.2, CT45A1 , CT45A2, CT45A3, CT45A4, CT45A5, CT45A6, CT47A1 , CT47A10, CT47A11 , CT47A2, CT47A3, CT47A4, CT47A5, CT47A6, CT47A7, CT47A8, CT47A9, CT47B1 , CT66, AA884595, CT69, BC040308, CT70, BI818097, CTAG1A, CTAG1 B, CTAG2, CTAGE-2, CTAGE1 , CTAGE5, CTCFL, CTNNA2, CXORF48, CXORF61 , CYCLIN A1 , DCAF12, DDX43, DDX53, DKKL1 , DMRT1 , DNAJB8, DPPA2, DSCR8, EDAG, NDR, ELOVL4, FAM133A, FAM46D, FATE1 , FBX039, FMR1 NB, FTHL17, GAGE1 , GAGE12B, GAGE12C, GAGE12D, GAGE12E, GAGE12F, GAGE12G, GAGE12H, GAGE12I, GAGE12J, GAGE13, GAGE2A, GAGE3, GAGE4, GAGE5, GAGE6, GAGE7, GAGE8, GOLGAGL2 FA, GPAT2, GPATCH2, HIWI, M IWI, PIWI, HORMAD1 , HORMAD2, HSPB9, IGSF11 , IL13RA2, IMP-3, JARID1 B, KIAA0100, LAGE-1 B, LDHC,
LEMD1, LIPI, LOC130576, LOC196993, LOC348120, LOC440934, LOC647107, LOC728137, LUZP4, LY6K, MAEL, MAGEA1, MAGEA10, MAGEA11 , MAGEA12, MAGEA2, MAGEA2B, MAGEA3, MAGEA4, MAGEA5, MAGEA6, MAGEA8, MAGEA9, MAGEA9B, LOC728269, MAGEB1 , MAGEB2, MAGEB3, MAGEB4, MAGEB5, MAGEB6, MAGEC1, MAGEC2, MAGEC3, MCAK, MMA1 B, M0RC1, MPH0SPH1 , NLRP4, N0L4, NR6A1, NXF2, NXF2B, NY-ESO-1 , 0DF1, 0DF2, 0DF3, 0DF4, 0IP5, OTOA, PAGE1, PAGE2, PAGE2B, PAGE3, PAGE4, PAGE5, PASD1, PBK, PEPP2, PIWIL2, PLAC1, POTEA, POTEB, POTEC, POTED, POTEE, POTEG, POTEH, PRAME, PRM1, PRM2, PRSS54, PRSS55, PTPN20A, RBM46, RGS22, ROPN1, RQCD1, SAGE1 , SEMG1, SLC06A1 , SPA17, SPACA3, SPAG1 , SPAG17, SPAG4, SPAG6, SPAG8, SPAG9, SPANXA1, SPANXA2, SPANXB1 , SPANXB2, SPANXC, SPANXD, SPANXE, SPANXN1 , SPANXN2, SPANXN3, SPANXN4, SPANXN5, SPATA19, SPEF2, SPINLW1, SP011 , SSX1, SSX2, SSX2B, SSX3, SSX4, SSX4B, SSX5, SSX6, SSX7, SSX9, SYCE1 , SYCP1, TAF7L, TAG, TDRD1, TDRD4, TDRD6, TEKT5, TEX101, TEX14, TEX15, TFDP3, THEG, TMEFF1, TMEFF2, TMEM108, TMPRSS12, TPPP2, TPTE, TSGA10, TSP50, TSPY1D, TSPY1 E, TSPY1 F, TSPY1G, TSPY1H, TSPY1 I, TSPY2, TSPY3, TSSK6, TTK, TULP2, VENTXP1, XAGE-3B, XAGE-4, RP11-167P23.2, XAGE1, XAGE1B, XAGE1C, XAGE1D, XAGE1 E, XAGE2, XAGE2B, CTD-2267G17.3, XAGE3, XAGE5, ZNF165, ZNF645, MART-1 /Melan-A, gp100, Dipeptidyl peptidase IV (DPPIV), adenosine deaminase-binding protein (ADAbp), cyclophilin b, Colorectal associated antigen (CRC)-0017- 1A/GA733, Carcinoembryonic Antigen (CEA) and its immunogenic epitopes CAP-1 and CAP-2, etv6, aml1 , Prostate Specific Antigen (PSA) and its immunogenic epitopes PSA-1, PSA-2, and PSA-3, prostate-specific membrane antigen (PSMA), T-cell receptor/CD3-zeta chain, RAGE, NAG, GnT-V, MUM-1 , CDK4, tyrosinase, p53, MUC family, HER2/neu, p21 ras, RCAS1, a-fetoprotein, E-cadherin, a- catenin, b-catenin and g-catenin, p120ctn, gp100 PmeM 17, cdc27, adenomatous polyposis coli protein (APC), fodrin, Connexin 37, Ig-idiotype, p15, gp75, GM2 and GD2 gangliosides, viral products such as human papilloma virus proteins, Smad family of tumor antigens, Imp-1, NA, EBV-encoded nuclear antigen (EBNA)-1, brain glycogen phosphorylase, SCP-1 CT-7, c-erbB-2, CD19, CD20, CD22, CD30, CD33, CD37, CD56, CD70, CD74, CD138, AGS16, MUC1 , GPNMB, Ep-CAM, PD-L1, PD-L2, PMSA,. In some embodiments, the antigens are human endogenous retroviral antigens. Illustrative antigens can also include antigens from human endogenous retroviruses which include, but are not limited to, epitopes derived from at least a portion of Gag, at least a portion of T at, at least a portion of Rev, a least a portion of Nef, and at least a portion of gp160.
[00120] Further, in some embodiments, the present vaccine protein fusions {e.g., gp96 fusions) provide for an adjuvant effect that further allows the immune system of a patient, when used in the various methods described herein, to be activated against a disease of interest.
[00121] Both prokaryotic and eukaryotic vectors can be used for expression of the vaccine protein (e.g., gp96-lg) and T cell costimulatory fusion proteins in the cell based therapy methods provided herein. Prokaryotic vectors include constructs based on E. coli sequences (see, e.g., Makrides, Microbiol Rev 1996, 60:512-538). Non-limiting examples of regulatory regions that can be used for expression in E. coli include lac, trp, Ipp, phoA, recA, tac, T3, T7 and APL. Non-limiting examples of prokaryotic expression vectors may include the Agt vector series such as Agt11 (Huynh et al., in "DNA Cloning Techniques, Vol. I: A Practical Approach,” 1984, (D. Glover, ed.), pp. 49-78, IRL Press, Oxford), and the pET vector series (Studier et al., Methods Enzymol 1990, 185:60-89). Prokaryotic host-vector systems cannot perform much of the post-translational processing of mammalian cells, however. Thus, eukaryotic host- vector systems may be particularly useful.
[00122] A variety of regulatory regions can be used for expression of the vaccine protein {e.g., gp96- Ig) and T cell costimulatory fusions in mammalian host cells. For example, the SV40 early and late promoters, the cytomegalovirus (CMV) immediate early promoter, and the Rous sarcoma virus long terminal repeat (RSV-LTR) promoter can be used. Inducible promoters that may be useful in mammalian cells include, without limitation, promoters associated with the metallothionein II gene, mouse mammary tumor virus glucocorticoid responsive long terminal repeats (MMTV-LTR), the b-interferon gene, and the hsp70 gene (see, Williams et al., Cancer Res 1989, 49:2735-42; and Taylor et al., Mol Cell Biol 1990, 10:165-75). Heat shock promoters or stress promoters also may be advantageous for driving expression of the fusion proteins in recombinant host cells.
[00123] In some embodiments, the present invention contemplates the use of inducible promoters capable of effecting high level of expression transiently in response to a cue. Illustrative inducible expression control regions include those comprising an inducible promoter that is stimulated with a cue such as a small molecule chemical compound. Particular examples can be found, for example, in U.S. Pat. Nos. 5,989,910, 5,935,934, 6,015,709, and 6,004,941 , each of which is incorporated herein by reference in its entirety.
[00124] Animal regulatory regions that exhibit tissue specificity and have been utilized in transgenic animals also can be used in tumor cells of a particular tissue type: the elastase I gene control region that is active in pancreatic acinar cells (Swift et al., Cell 1984, 38:639-646; Ornitz et al., Cold Spring Harbor Symp Quant Biol 1986, 50:399-409; and MacDonald, Hepatology 1987, 7:425-515); the insulin gene control region that is active in pancreatic beta cells (Hanahan, Nature 1985, 315:115-122), the immunoglobulin gene control region that is active in lymphoid cells (Grosschedl et al., Cell 1984, 38:647- 658; Adames et al., Nature 1985, 318:533-538; and Alexander et al., Mol Cell Biol 1987, 7:1436-1444), the mouse mammary tumor virus control region that is active in testicular, breast, lymphoid and mast
cells (Leder et al., Cell 1986, 45:485-495), the albumin gene control region that is active in liver (Pinkert et al., Genes Devel, 1987, 1 :268-276), the alpha-fetoprotein gene control region that is active in liver (Krumlauf et al., Mol Cell Biol 1985, 5:1639-1648; and Hammer et al., Science 1987, 235:53-58); the alpha 1-antitrypsin gene control region that is active in liver (Kelsey et al., Genes Devel 1987, 1 :161-171), the beta-globin gene control region that is active in myeloid cells (Mogram et al., Nature 1985, 315:338- 340; and Kollias et al., Cell 1986, 46:89-94); the myelin basic protein gene control region that is active in oligodendrocyte cells in the brain (Readhead et al., Cell 1987, 48:703-712); the myosin light chain-2 gene control region that is active in skeletal muscle (Sani, Nature 1985, 314:283-286), and the gonadotropic releasing hormone gene control region that is active in the hypothalamus (Mason et al., Science 1986, 234:1372-1378).
[00125] An expression vector also can include transcription enhancer elements, such as those found in SV40 virus, Hepatitis B virus, cytomegalovirus, immunoglobulin genes, metallothionein, and b-actin (see, Bittner et al., Meth Enzymol 1987, 153:516-544; and Gorman, Curr Op Biotechnol 1990, 1 :36-47). In addition, an expression vector can contain sequences that permit maintenance and replication of the vector in more than one type of host cell, or integration of the vector into the host chromosome. Such sequences include, without limitation, to replication origins, autonomously replicating sequences (ARS), centromere DNA, and telomere DNA.
[00126] In addition, an expression vector can contain one or more selectable or screenable marker genes for initially isolating, identifying, or tracking host cells that contain DNA encoding fusion proteins as described herein. For long term, high yield production of gp96-lg and T cell costimulatory fusion proteins, stable expression in mammalian cells can be useful. A number of selection systems can be used for mammalian cells. For example, the Herpes simplex virus thymidine kinase (Wigler et al., Cell 1977, 11 :223), hypoxanthine-guanine phosphoribosyltransferase (Szybalski and Szybalski, Proc Natl Acad Sci USA 1962, 48:2026), and adenine phosphoribosyltransferase (Lowy et al., Cell 1980, 22:817) genes can be employed in tk-, hgprt-, or aprt- cells, respectively. In addition, antimetabolite resistance can be used as the basis of selection for dihydrofolate reductase (dhfr), which confers resistance to methotrexate (Wigler et al., Proc Natl Acad Sci USA 1980, 77:3567; O'Hare et al., Proc Natl Acad Sci USA 1981 , 78:1527); gpt, which confers resistance to mycophenolic acid (Mulligan and Berg, Proc Natl Acad Sci USA 1981 , 78:2072); neomycin phosphotransferase (neo), which confers resistance to the aminoglycoside G-418 (Colberre-Garapin et al., J Mol Biol 1981 , 150:1); and hygromycin phosphotransferase (hyg), which confers resistance to hygromycin (Santerre et al., Gene 1984, 30:147). Other selectable markers such as histidinol and Zeocin™ also can be used.
[00127] Useful mammalian host cells include, without limitation, cells derived from humans, monkeys,
and rodents (see, for example, Kriegler in "Gene Transfer and Expression: A Laboratory Manual,” 1990, New York, Freeman & Co.). These include monkey kidney cell lines transformed by SV40 {e.g., COS-7, AT CC CRL 1651 ); human embryonic kidney lines (e.g., 293, 293-EBNA, or 293 cells subcloned for growth in suspension culture, Graham et al., J Gen Virol 1977, 36:59); baby hamster kidney cells (e.g., BHK, ATCC CCL 10); Chinese hamster ovary-cells-DHFR (e.g., CHO, Urlaub and Chasin, Proc Natl Acad Sci USA 1980, 77:4216); mouse sertoli cells (Mather, Biol Reprod 1980, 23:243-251 ); mouse fibroblast cells (e.g., NIH-3T3), monkey kidney cells (e.g., CV1 ATCC CCL 70); African green monkey kidney cells, (e.g., VERO-76, ATCC CRL-1587); human cervical carcinoma cells (e.g., HELA, ATCC CCL 2); canine kidney cells (e.g., MDCK, ATCC CCL 34); buffalo rat liver cells (e.g., BRL 3A, ATCC CRL 1442); human lung cells (e.g., W138, ATCC CCL 75); human liver cells (e.g., Hep G2, FIB 8065); and mouse mammary tumor cells (e.g., MMT 060562, ATCC CCL51). Illustrative cancer cell types for expressing the fusion proteins described herein include mouse fibroblast cell line, NIH3T3, mouse Lewis lung carcinoma cell line, LLC, mouse mastocytoma cell line, P815, mouse lymphoma cell line, EL4 and its ovalbumin transfectant, E.G7, mouse melanoma cell line, B16F10, mouse fibrosarcoma cell line, MC57, human small cell lung carcinoma cell lines, SCLC#2 and SCLC#7, human lung adenocarcinoma cell line, e.g., AD100, and human prostate cancer cell line, e.g., PC-3.
[00128] A number of viral-based expression systems also can be used with mammalian cells to produce gp96-lg and T cell costimulatory fusion proteins. Vectors using DNA virus backbones have been derived from simian virus 40 (SV40) (Flamer et al., Cell 1979, 17:725), adenovirus (Van Doren et al., Mol Cell Biol 1984, 4:1653), adeno-associated virus (McLaughlin et al., J Virol 1988, 62:1963), and bovine papillomas virus (Zinn et al., Proc Natl Acad Sci USA 1982, 79:4897). When an adenovirus is used as an expression vector, the donor DNA sequence may be ligated to an adenovirus transcription/translation control complex, e.g., the late promoter and tripartite leader sequence. This fusion gene may then be inserted in the adenovirus genome by in vitro or in vivo recombination. Insertion in a non-essential region of the viral genome (e.g., region E1 or E3) can result in a recombinant virus that is viable and capable of expressing heterologous products in infected hosts. (See, e.g., Logan and Shenk, Proc Natl Acad Sci USA 1984, 81 :3655-3659).
[00129] Bovine papillomavirus (BPV) can infect many higher vertebrates, including man, and its DNA replicates as an episome. A number of shuttle vectors have been developed for recombinant gene expression which exist as stable, multicopy (20-300 copies/cell) extrachromosomal elements in mammalian cells. Typically, these vectors contain a segment of BPV DNA (the entire genome or a 69% transforming fragment), a promoter with a broad host range, a polyadenylation signal, splice signals, a selectable marker, and "poisonless” plasmid sequences that allow the vector to be propagated in E. coli.
Following construction and amplification in bacteria, the expression gene constructs are transfected into cultured mammalian cells by, for example, calcium phosphate coprecipitation. For those host cells that do not manifest a transformed phenotype, selection of transformants is achieved by use of a dominant selectable marker, such as histidinol and G418 resistance.
[00130] Alternatively, the vaccinia 7.5K promoter can be used. (See, e.g., Mackett et al., Proc Natl Acad Sci USA 1982, 79:7415-7419; Mackett et al., J Virol 1984, 49:857-864; and Panicali et al., Proc Natl Acad Sci USA 1982, 79:4927-4931.) In cases where a human host cell is used, vectors based on the Epstein- Barr virus (EBV) origin (OriP) and EBV nuclear antigen 1 (EBNA-1 ; a trans-acting replication factor) can be used. Such vectors can be used with a broad range of human host cells, e.g., EBO-pCD (Spickofsky et al., DNA Prot Eng Tech 1990, 2:14-18); pDR2 and ADR2 (available from Clontech Laboratories).
[00131] Gp96-lg and T cell costimulatory fusion proteins also can be made with retrovirus-based expression systems. Retroviruses, such as Moloney murine leukemia virus, can be used since most of the viral gene sequence can be removed and replaced with exogenous coding sequence while the missing viral functions can be supplied in trans. In contrast to transfection, retroviruses can efficiently infect and transfer genes to a wide range of cell types including, for example, primary hematopoietic cells. Moreover, the host range for infection by a retroviral vector can be manipulated by the choice of envelope used for vector packaging.
[00132] For example, a retroviral vector can comprise a 5' long terminal repeat (LTR), a 3' LTR, a packaging signal, a bacterial origin of replication, and a selectable marker. The gp96-lg fusion protein coding sequence, for example, can be inserted into a position between the 5' LTR and 3' LTR, such that transcription from the 5' LTR promoter transcribes the cloned DNA. The 5' LTR contains a promoter {e.g., an LTR promoter), an R region, a U5 region, and a primer binding site, in that order. Nucleotide sequences of these LTR elements are well known in the art. A heterologous promoter as well as multiple drug selection markers also can be included in the expression vector to facilitate selection of infected cells. See, McLauchlin et al., Prog Nucleic Acid Res Mol Biol 1990, 38:91-135; Morgenstern et al., Nucleic Acid Res 1990, 18:3587-3596; Choulika et al., J Virol 1996, 70:1792-1798; Boesen et al., Biotherapy 1994, 6:291-302; Salmons and Gunzberg, Pluman Gene Ther 1993, 4:129-141 ; and Grossman and Wilson, Curr Opin Genet Devel 1993, 3:110-114.
[00133] Any of the cloning and expression vectors described herein may be synthesized and assembled from known DNA sequences using techniques that are known in the art. The regulatory regions and enhancer elements can be of a variety of origins, both natural and synthetic. Some vectors and host cells may be obtained commercially. Non-limiting examples of useful vectors are described in Appendix 5 of Current Protocols in Molecular Biology, 1988, ed. Ausubel et al., Greene Publish. Assoc. & Wiley
Interscience, which is incorporated herein by reference; and the catalogs of commercial suppliers such as Clontech Laboratories, Stratagene Inc., and Invitrogen, Inc.
Methods of Treating
[00134] Cell based therapies can be used in administration to a subject {e.g., a research animal or a mammal, such as a human, having a clinical condition such as cancer or an infection). For example, the cell based therapy can be administered to a subject for the treatment of cancer or infection. Thus, this document provides methods for treating clinical conditions such as cancer or infection with the expression vectors provided herein. The infection can be, for example, an acute infection or a chronic infection. In some embodiments, the infection can be an infection by hepatitis C virus, hepatitis B virus, human immunodeficiency virus, or malaria. The methods can include administering to a subject the cell based therapies under conditions wherein the progression or a symptom of the clinical condition in the subject is reduced in a therapeutic manner.
[00135] In various embodiments, the present invention pertains to cancers and/or tumors; for example, the treatment or prevention of cancers and/or tumors. Cancers or tumors refer to an uncontrolled growth of cells and/or abnormal increased cell survival and/or inhibition of apoptosis which interferes with the normal functioning of the bodily organs and systems. Included are benign and malignant cancers, polyps, hyperplasia, as well as dormant tumors or micrometastases. Also, included are cells having abnormal proliferation that is not impeded by the immune system {e.g., virus infected cells). The cancer may be a primary cancer or a metastatic cancer. The primary cancer may be an area of cancer cells at an originating site that becomes clinically detectable, and may be a primary tumor. In contrast, the metastatic cancer may be the spread of a disease from one organ or part to another non-adjacent organ or part. The metastatic cancer may be caused by a cancer cell that acquires the ability to penetrate and infiltrate surrounding normal tissues in a local area, forming a new tumor, which may be a local metastasis. The cancer may also be caused by a cancer cell that acquires the ability to penetrate the walls of lymphatic and/or blood vessels, after which the cancer cell is able to circulate through the bloodstream (thereby being a circulating tumor cell) to other sites and tissues in the body. The cancer may be due to a process such as lymphatic or hematogeneous spread. The cancer may also be caused by a tumor cell that comes to rest at another site, re-penetrates through the vessel or walls, continues to multiply, and eventually forms another clinically detectable tumor. The cancer may be this new tumor, which may be a metastatic (or secondary) tumor.
[00136] The cancer may be caused by tumor cells that have metastasized, which may be a secondary or metastatic tumor. The cells of the tumor may be like those in the original tumor. As an example, if a
breast cancer or colon cancer metastasizes to the liver, the secondary tumor, while present in the liver, is made up of abnormal breast or colon cells, not of abnormal liver cells. The tumor in the liver may thus be a metastatic breast cancer or a metastatic colon cancer, not liver cancer.
[00137] The cancer may have an origin from any tissue. The cancer may originate from melanoma, colon, breast, or prostate, and thus may be made up of cells that were originally skin, colon, breast, or prostate, respectively. The cancer may also be a hematological malignancy, which may be lymphoma. The cancer may invade a tissue such as liver, lung, bladder, or intestinal.
[00138] Illustrative cancers that may be treated include, but are not limited to, carcinomas, e.g. various subtypes, including, for example, adenocarcinoma, basal cell carcinoma, squamous cell carcinoma, and transitional cell carcinoma), sarcomas (including, for example, bone and soft tissue), leukemias (including, for example, acute myeloid, acute lymphoblastic, chronic myeloid, chronic lymphocytic, and hairy cell), lymphomas and myelomas (including, for example, Hodgkin and non-Hodgkin lymphomas, light chain, non-secretory, MGUS, and plasmacytomas), and central nervous system cancers (including, for example, brain (e.g. gliomas {e.g., astrocytoma, oligodendroglioma, and ependymoma), meningioma, pituitary adenoma, and neuromas, and spinal cord tumors (e.g., meningiomas and neurofibroma).
[00139] Representative cancers and/or tumors of the present invention include, but are not limited to, a basal cell carcinoma, biliary tract cancer; bladder cancer; bone cancer; brain and central nervous system cancer; breast cancer; cancer of the peritoneum; cervical cancer; choriocarcinoma; colon and rectum cancer; connective tissue cancer; cancer of the digestive system; endometrial cancer; esophageal cancer; eye cancer; cancer of the head and neck; gastric cancer (including gastrointestinal cancer); glioblastoma; hepatic carcinoma; hepatoma; intra-epithelial neoplasm; kidney or renal cancer; larynx cancer; leukemia; liver cancer; lung cancer (e.g., small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous carcinoma of the lung); melanoma; myeloma; neuroblastoma; oral cavity cancer (lip, tongue, mouth, and pharynx); ovarian cancer; pancreatic cancer; prostate cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; cancer of the respiratory system; salivary gland carcinoma; sarcoma; skin cancer; squamous cell cancer; stomach cancer; testicular cancer; thyroid cancer; uterine or endometrial cancer; cancer of the urinary system; vulval cancer; lymphoma including Hodgkin's and non-Hodgkin's lymphoma, as well as B-cell lymphoma (including low grade/follicular non-Hodgkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade/follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small non-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's Macroglobulinemia; chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); Hairy cell leukemia; chronic myeloblastic leukemia; as well as other
carcinomas and sarcomas; and post-transplant lymphoproliferative disorder (PTLD), as well as abnormal vascular proliferation associated with phakomatoses, edema (such as that associated with brain tumors), and Meigs' syndrome.
[00140] In some aspects, the present fusions are used to eliminate intracellular pathogens. In some aspects, the present fusions are used to treat one or more infections. In some embodiments, the present fusion proteins are used in methods of treating viral infections (including, for example, HIV and HCV), parasitic infections (including, for example, malaria), and bacterial infections. In various embodiments, the infections induce immunosuppression. For example, HIV infections often result in immunosuppression in the infected subjects. Accordingly, as described elsewhere herein, the treatment of such infections may involve, in various embodiments, modulating the immune system with the present fusion proteins to favor immune stimulation over immune inhibition. Alternatively, the present invention provides methods for treating infections that induce immunoactivation. For example, intestinal helminth infections have been associated with chronic immune activation. In these embodiments, the treatment of such infections may involve modulating the immune system with the present fusion proteins to favor immune inhibition over immune stimulation.
[00141] In various embodiments, the present invention provides methods of treating viral infections including, without limitation, acute or chronic viral infections, for example, of the respiratory tract, of papilloma virus infections, of herpes simplex virus (HSV) infection, of human immunodeficiency virus (HIV) infection, and of viral infection of internal organs such as infection with hepatitis viruses. In some embodiments, the viral infection is caused by a virus of family Flaviviridae. In some embodiments, the virus of family Flaviviridae is selected from Yellow Fever Virus, West Nile virus, Dengue virus, Japanese Encephalitis Virus, St. Louis Encephalitis Virus, and Hepatitis C Virus. In other embodiments, the viral infection is caused by a virus of family Picornaviridae, e.g., poliovirus, rhinovirus, coxsackievirus. In other embodiments, the viral infection is caused by a member of Orthomyxoviridae, e.g., an influenza virus. In other embodiments, the viral infection is caused by a member of Retroviridae, e.g., a lentivirus. In other embodiments, the viral infection is caused by a member of Paramyxoviridae, e.g., respiratory syncytial virus, a human parainfluenza virus, rubulavirus {e.g., mumps virus), measles virus, and human metapneumovirus. In other embodiments, the viral infection is caused by a member of Bunyaviridae, e.g., hantavirus. In other embodiments, the viral infection is caused by a member of Reoviridae, e.g., a rotavirus.
[00142] In various embodiments, the present invention provides methods of treating parasitic infections such as protozoan or helminths infections. In some embodiments, the parasitic infection is by a protozoan parasite. In some embodiments, the oritiziab parasite is selected from intestinal protozoa, tissue protozoa,
or blood protozoa. Illustrative protozoan parasites include, but are not limited to, Entamoeba hystolytica, Giardia lamblia, Cryptosporidium muris, Trypanosomatida gambiense, Trypanosomatida rhodesiense, Trypanosomatida crusi, Leishmania mexicana, Leishmania braziliensis, Leishmania tropica, Leishmania donovani, Toxoplasma gondii, Plasmodium vivax, Plasmodium ovale, Plasmodium malariae, Plasmodium falciparum, Trichomonas vaginalis, and Histomonas meleagridis. In some embodiments, the parasitic infection is by a helminthic parasite such as nematodes {e.g., Adenophorea). In some embodiments, the parasite is selected from Secementea {e.g., Trichuris trichiura, Ascaris lumbricoides, Enterobius vermicularis, Ancylostoma duodenale, Necator americanus, Strongyloides stercoralis, Wuchereria bancrofti, Dracunculus medinensis). In some embodiments, the parasite is selected from trematodes (e.g. blood flukes, liver flukes, intestinal flukes, and lung flukes). In some embodiments, the parasite is selected from: Schistosoma mansoni, Schistosoma haematobium, Schistosoma japonicum, Fasciola hepatica, Fasciola gigantica, Heterophyes heterophyes, Paragonimus westermani. In some embodiments, the parasite is selected from cestodes (e.g., Taenia solium, Taenia saginata, Hymenolepis nana, Echinococcus granulosus).
[00143] In various embodiments, the present invention provides methods of treating bacterial infections. In various embodiments, the bacterial infection is by a gram-positive bacterium, gram-negative bacteria, aerobic and/or anaerobic bacteria. In various embodiments, the bacteria is selected from, but not limited to, Staphylococcus, Lactobacillus, Streptococcus, Sarcina, Escherichia, Enterobacter, Klebsiella, Pseudomonas, Acinetobacter, Mycobacterium, Proteus, Campylobacter, Citrobacter, Nisseria, Baccillus, Bacteroides, Peptococcus, Clostridium, Salmonella, Shigella, Serratia, Haemophilus, Brucella and other organisms. In some embodiments, the bacteria is selected from, but not limited to, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas acidovorans, Pseudomonas alcaligenes, Pseudomonas putida, Stenotrophomonas maltophilia, Burkholderia cepacia, Aeromonas hydrophilia, Escherichia coli, Citrobacter freundii, Salmonella typhimurium, Salmonella typhi, Salmonella paratyphi, Salmonella enteritidis, Shigella dysenteriae, Shigella flexneri, Shigella sonnei, Enterobacter cloacae, Enterobacter aerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Serratia marcescens, Francisella tularensis, Morganella morganii, Proteus mirabilis, Proteus vulgaris, Providencia alcalifaciens, Providencia rettgeri, Providencia stuartii, Acinetobacter baumannii, Acinetobacter calcoaceticus, Acinetobacter haemolyticus, Yersinia enterocolitica, Yersinia pestis, Yersinia pseudotuberculosis, Yersinia intermedia, Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus haemolyticus, Haemophilus parahaemolyticus, Haemophilus ducreyi, Pasteurella multocida, Pasteurella haemolytica, Branhamella catarrhalis, Helicobacter pylori, Campylobacter fetus, Campylobacter jejuni, Campylobacter coli, Borrelia
burgdorferi, Vibrio cholerae, Vibrio parahaemolyticus, Legionella pneumophila, Listeria monocytogenes, Neisseria gonorrhoeae, Neisseria meningitidis, Kingella, Moraxella, Gardnerella vaginalis, Bacteroides fragilis, Bacteroides distasonis, Bacteroides 3452A homology group, Bacteroides vulgatus, Bacteroides ovalus, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bacteroides eggerthii, Bacteroides splanchnicus, Clostridium difficile, Mycobacterium tuberculosis, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium leprae, Corynebacterium diphtheriae, Corynebacterium ulcerans, Streptococcus pneumoniae, Streptococcus agalactiae, Streptococcus pyogenes, Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Staphylococcus intermedius, Staphylococcus hyicus subsp. hyicus, Staphylococcus haemolyticus, Staphylococcus hominis, or Staphylococcus saccharolyticus. The cell based therapy to be administered can be admixed, encapsulated, conjugated or otherwise associated with other molecules, molecular structures, or mixtures of compounds such as, for example, liposomes, receptor or cell targeted molecules, or oral, topical or other formulations for assisting in uptake, distribution and/or absorption. The cell based therapy to be administered can be in combination with a pharmaceutically acceptable carrier.
[00144] This present disclosure therefore also provides compositions containing cell based therapies described herein, in combination with a physiologically and pharmaceutically acceptable carrier. The physiologically and pharmaceutically acceptable carrier can be include any of the well-known components useful for immunization. The carrier can facilitate or enhance an immune response to an antigen administered in a vaccine. The cell formulations can contain buffers to maintain a preferred pH range, salts or other components that present an antigen to an individual in a composition that stimulates an immune response to the antigen. The physiologically acceptable carrier also can contain one or more adjuvants that enhance the immune response to an antigen. Pharmaceutically acceptable carriers include, for example, pharmaceutically acceptable solvents, suspending agents, or any other pharmacologically inert vehicles for delivering compounds to a subject. Pharmaceutically acceptable carriers can be liquid or solid, and can be selected with the planned manner of administration in mind so as to provide for the desired bulk, consistency, and other pertinent transport and chemical properties, when combined with one or more therapeutic compounds and any other components of a given pharmaceutical composition. Typical pharmaceutically acceptable carriers include, without limitation: water, saline solution, binding agents {e.g., polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers {e.g., lactose or dextrose and other sugars, gelatin, or calcium sulfate), lubricants {e.g., starch, polyethylene glycol, or sodium acetate), disintegrates (e.g., starch or sodium starch glycolate), and wetting agents (e.g., sodium lauryl sulfate). Compositions can be formulated for subcutaneous, intramuscular, or intradermal administration, or in any manner acceptable for immunization.
[00145] An adjuvant refers to a substance which, when added to an immunogenic agent such as a tumor cell expressing secreted vaccine protein {e.g., gp96-lg) and T cell costimulatory fusion polypeptides, nonspecifically enhances or potentiates an immune response to the agent in the recipient host upon exposure to the mixture. Adjuvants can include, for example, oil-in-water emulsions, water-in oil emulsions, alum (aluminum salts), liposomes and microparticles, such as, polysytrene, starch, polyphosphazene and polylactide/polyglycosides.
[00146] Adjuvants can also include, for example, squalene mixtures (SAF-I), muramyl peptide, saponin derivatives, mycobacterium cell wall preparations, monophosphoryl lipid A, mycolic acid derivatives, nonionic block copolymer surfactants, Quil A, cholera toxin B subunit, polyphosphazene and derivatives, and immunostimulating complexes (ISCOMs) such as those described by Takahashi et al., Nature 1990, 344:873-875. For veterinary use and for production of antibodies in animals, mitogenic components of Freund's adjuvant (both complete and incomplete) can be used. In humans, Incomplete Freund's Adjuvant (IFA) is a useful adjuvant. Various appropriate adjuvants are well known in the art (see, for example, Warren and Chedid, CRC Critical Reviews in Immunology 1988, 8:83; and Allison and Byars, in Vaccines: New Approaches to Immunological Problems, 1992, Ellis, ed., Butterworth-Fleinemann, Boston). Additional adjuvants include, for example, bacille Calmett-Guerin (BCG), DETOX (containing cell wall skeleton of Mycobacterium phlei (CWS) and monophosphoryl lipid A from Salmonella minnesota (MPL)), and the like (see, for example, Floover et al., J Clin Oncol 1993, 1 1 :390; and Woodlock et al., J Immunother 1999, 22:251 -259).
[00147] In some embodiments, a cell secreting a T cell costimulatory fusion protein {e.g., OX40L-lg) can be administered to a subject at about 100,000 cells, about 150,000 cells, about 200,000 cells, about 250,000 cells, about 300,000 cells, about 350,000 cells, about 400,000 cells, about 450,000 cells, about
500,000 cells, about 550,000 cells, about 600,000 cells, about 650,000 cells, about 700,000 cells, about
750,000 cells, about 800,000 cells, about 850,000 cells, about 900,000 cells, about 950,000 cells, about
1 million cells, about 1.5 million cells, about 2 million cells, about 2.5 million cells, about 3 million cells, about 3.5 million cells, about 4 million cells, about 4.5 million cells, about 6 million cells, about 6.5 million cells, about 7 million cells, about 7.5 million cells, about 8 million cells, about 8.5 million cells, about 9 million cells, about 9.5 million cells, or about 10 million cells.
[00148] In some embodiments, a cell secreting a vaccine protein (e.g., gp96-lg) can be administered to a subject at about 100,000 cells, about 150,000 cells, about 200,000 cells, about 250,000 cells, about
300,000 cells, about 350,000 cells, about 400,000 cells, about 450,000 cells, about 500,000 cells, about
550,000 cells, about 600,000 cells, about 650,000 cells, about 700,000 cells, about 750,000 cells, about
800,000 cells, about 850,000 cells, about 900,000 cells, about 950,000 cells, about 1 million cells, about
1.5 million cells, about 2 million cells, about 2.5 million cells, about 3 million cells, about 3.5 million cells, about 4 million cells, about 4.5 million cells, about 6 million cells, about 6.5 million cells, about 7 million cells, about 7.5 million cells, about 8 million cells, about 8.5 million cells, about 9 million cells, about 9.5 million cells, or about 10 million cells.
[00149] In some embodiments, a fixed dose of a cell secreting a vaccine protein is 1 x 107 cells.
[00150] In some embodiments, a fixed dose of a cell secreting a T cell costimulatory fusion protein {e.g., OX40L-lg) is 1 x 107 cells.
[00151] In some embodiments, about a cell secreting a T cell costimulatory fusion protein {e.g., OX40L- Ig) can be administered to a subject at about 0.1 -1.1 x 107 cells, about 0.2- 1 .1 x 107 cells, about 0.3- 1.1 x 107 cells, about 0.4- 1 .1 x 107 cells, about 0.5- 1 .1 x 107 cells, about 0.6- 1.1 x 107 cells, about 0.7- 1 .1 x 107 cells, about 0.8- 1.1 x 107 cells, about or 0.9- 1 .1 x 107 cells, about 0.1-2.1 x 107 cells, about 0.2-
2.1 x 107 cells, about 0.3- 2.1 x 107 cells, about 0.4- 2.1 x 107 cells, about 0.5- 2.1 x 107 cells, about 0.6-
2.1 x 107 cells, about 0.7- 2.1 x 107 cells, about 0.8- 2.1 x 107 cells, about 0.9- 2.1 x 107 cells, about 0.1 -
3.1 x 107 cells, about 0.2- 3.1 x 107 cells, about 0.3- 3.1 x 107 cells, about 0.4- 3.1 x 107 cells, about 0.5-
3.1 x 107 cells, about 0.6- 3.1 x 107 cells, about 0.7- 3.1 x 107 cells, about 0.8- 3.1 x 107 cells, about 0.9-
3.1 x 107 cells, about 0.1-4.1 x 107 cells, about 0.2- 4.1 x 107 cells, about 0.3- 4.1 x 107 cells, about 0.4-
4.1 x 107 cells, about 0.5- 4.1 x 107 cells, about 0.6- 4.1 x 107 cells, about 0.7- 4.1 x 107 cells, about 0.8-
4.1 x 107 cells, about 0.9- 4.1 x 107 cells, about 0.1 -5.1 x 107 cells, about 0.2- 5.1 x 107 cells, about 0.3-
5.1 x 107 cells, about 0.4- 5.1 x 107 cells, about 0.5- 5.1 x 107 cells, about 0.6- 5.1 x 107 cells, about 0.7-
5.1 x 107 cells, about 0.8- 5.1 x 107 cells, about 0.9- 5.1 x 107 cells, about 0.1 -6.1 x 107 cells, about 0.2-
6.1 x 107 cells, about 0.3- 6.1 x 107 cells, about 0.4- 6.1 x 107 cells, about 0.5- 6.1 x 107 cells, about 0.6-
6.1 x 107 cells, about 0.7- 6.1 x 107 cells, about 0.8- 6.1 x 107 cells, about 0.9- 6.1 x 107 cells, about 0.1 -
7.1 x 107 cells, about 0.2- 7.1 x 107 cells, about 0.3- 7.1 x 107 cells, about 0.4- 7.1 x 107 cells, about 0.5-
7.1 x 107 cells, about 0.6- 7.1 x 107 cells, about 0.7- 7.1 x 107 cells, about 0.8- 7.1 x 107 cells, about 0.9-
7.1 x 107 cells, about 0.1-8.1 x 107 cells, about 0.2- 8.1 x 107 cells, about 0.3- 8.1 x 107 cells, about 0.4-
8.1 x 107 cells, about 0.5- 8.1 x 107 cells, about 0.6- 8.1 x 107 cells, about 0.7- 8.1 x 107 cells, about 0.8-
8.1 x 107 cells, about 0.9- 8.1 x 107 cells, about 0.1 -9.1 x 107 cells, about 0.2- 9.1 x 107 cells, about 0.3-
9.1 x 107 cells, about 0.4- 9.1 x 107 cells, about 0.5- 9.1 x 107 cells, about 0.6- 9.1 x 107 cells, about 0.7-
9.1 x 107 cells, about 0.8- 9.1 x 107 cells, or about 0.9- 9.1 x 107 cells.
[00152] In some embodiments, a cell secreting a vaccine protein (e.g., gp96-lg) can be administered to a subject at about 0.1-1.1 x 107 cells, about 0.2- 1 .1 x 107 cells, about 0.3- 1 .1 x 107 cells, about 0.4- 1 .1 x 107 cells, about 0.5- 1 .1 x 107 cells, about 0.6- 1 .1 x 107 cells, about 0.7- 1.1 x 107 cells, about 0.8- 1 .1 x 107 cells, about or 0.9- 1 .1 x 107 cells, about 0.1-2.1 x 107 cells, about 0.2- 2.1 x 107 cells, about 0.3-
2.1 x 107 cells, about 0.4- 2.1 x 107 cells, about 0.5- 2.1 x 107 cells, about 0.6- 2.1 x 107 cells, about 0.7-
2.1 x 107 cells, about 0.8- 2.1 x 107 cells, about 0.9- 2.1 x 107 cells, about 0.1 -3.1 x 107 cells, about 0.2-
3.1 x 107 cells, about 0.3- 3.1 x 107 cells, about 0.4- 3.1 x 107 cells, about 0.5- 3.1 x 107 cells, about 0.6-
3.1 x 107 cells, about 0.7- 3.1 x 107 cells, about 0.8- 3.1 x 107 cells, about 0.9- 3.1 x 107 cells, about 0.1 -
4.1 x 107 cells, about 0.2- 4.1 x 107 cells, about 0.3- 4.1 x 107 cells, about 0.4- 4.1 x 107 cells, about 0.5-
4.1 x 107 cells, about 0.6- 4.1 x 107 cells, about 0.7- 4.1 x 107 cells, about 0.8- 4.1 x 107 cells, about 0.9-
4.1 x 107 cells, about 0.1-5.1 x 107 cells, about 0.2- 5.1 x 107 cells, about 0.3- 5.1 x 107 cells, about 0.4-
5.1 x 107 cells, about 0.5- 5.1 x 107 cells, about 0.6- 5.1 x 107 cells, about 0.7- 5.1 x 107 cells, about 0.8-
5.1 x 107 cells, about 0.9- 5.1 x 107 cells, about 0.1 -6.1 x 107 cells, about 0.2- 6.1 x 107 cells, about 0.3-
6.1 x 107 cells, about 0.4- 6.1 x 107 cells, about 0.5- 6.1 x 107 cells, about 0.6- 6.1 x 107 cells, about 0.7-
6.1 x 107 cells, about 0.8- 6.1 x 107 cells, about 0.9- 6.1 x 107 cells, about 0.1 -7.1 x 107 cells, about 0.2-
7.1 x 107 cells, about 0.3- 7.1 x 107 cells, about 0.4- 7.1 x 107 cells, about 0.5- 7.1 x 107 cells, about 0.6-
7.1 x 107 cells, about 0.7- 7.1 x 107 cells, about 0.8- 7.1 x 107 cells, about 0.9- 7.1 x 107 cells, about 0.1 -
8.1 x 107 cells, about 0.2- 8.1 x 107 cells, about 0.3- 8.1 x 107 cells, about 0.4- 8.1 x 107 cells, about 0.5-
8.1 x 107 cells, about 0.6- 8.1 x 107 cells, about 0.7- 8.1 x 107 cells, about 0.8- 8.1 x 107 cells, about 0.9-
8.1 x 107 cells, about 0.1-9.1 x 107 cells, about 0.2- 9.1 x 107 cells, about 0.3- 9.1 x 107 cells, about 0.4-
9.1 x 107 cells, about 0.5- 9.1 x 107 cells, about 0.6- 9.1 x 107 cells, about 0.7- 9.1 x 107 cells, about 0.8-
9.1 x 107 cells, or about 0.9- 9.1 x 107 cells.
[00153] In some embodiments, the cell based therapy can be administered to a subject one or more times ( e.g once, twice, two to four times, three to five times, five to eight times, six to ten times, eight to 12 times, or more than 12 times). The cell based therapy as provided herein can be administered one or more times per day, one or more times per week, every other week, one or more times per month, once every two to three months, once every three to six months, or once every six to 12 months. The cell based therapy can be administered over any suitable period of time, such as a period from about 1 day to about 12 months. In some embodiments, for example, the period of administration can be from about 1 day to 90 days; from about 1 day to 60 days; from about 1 day to 30 days; from about 1 day to 20 days; from about 1 day to 10 days; from about 1 day to 7 days. In some embodiments, the period of administration can be from about 1 week to 50 weeks; from about 1 week to 50 weeks; from about 1 week to 40 weeks; from about 1 week to 30 weeks; from about 1 week to 24 weeks; from about 1 week to 20 weeks; from about 1 week to 16 weeks; from about 1 week to 12 weeks; from about 1 week to 8 weeks; from about 1 week to 4 weeks; from about 1 week to 3 weeks; from about 1 week to 2 weeks; from about 2 weeks to 3 weeks; from about 2 weeks to 4 weeks; from about 2 weeks to 6 weeks; from about 2 weeks to 8 weeks; from about 3 weeks to 8 weeks; from about 3 weeks to 12 weeks; or from
about 4 weeks to 20 weeks or any weekly increment of time in between.
[00154] In some embodiments, after an initial dose (sometimes referred to as a "priming” dose) of a cell based therapy has been administered and a maximal antigen-specific immune response has been achieved, one or more boosting doses of the cell based therapy as provided herein can be administered. For example, a boosting dose can be administered about 10 to 30 days, about 15 to 35 days, about 20 to 40 days, about 25 to 45 days, or about 30 to 50 days after a priming dose.
[00155] In some embodiments, a secreatable vaccine protein {e.g., gp96-lg) and the T cell costimulatory fusion protein {e.g., a cell secreting OX40L-lg), are administered 1 minute apart, 10 minutes apart, 30 minutes apart, less than 1 hour apart, 1 hour apart, 1 hour to 2 hours apart, 2 hours to 3 hours apart, 3 hours to 4 hours apart, 4 hours to 5 hours apart, 5 hours to 6 hours apart, 6 hours to 7 hours apart, 7 hours to 8 hours apart, 8 hours to 9 hours apart, 9 hours to 10 hours apart, 10 hours to 1 1 hours apart, 11 hours to 12 hours apart, 1 day apart, 2 days apart, 3 days apart, 4 days apart, 5 days apart, 6 days apart, 1 week apart, 2 weeks apart, 3 weeks apart, or 4 weeks apart.
[00156] In some embodiments, a regimen is provided in which a first treatment of a cell secreting a T cell costimulatory fusion protein (e.g., OX40L-lg) is administered and subsequently a second treatment of a cell secreting a T cell costimulatory fusion protein (e.g., OX40L-lg) is administered and a cell secreting a vaccine protein [e.g., gp96-lg) is administered. For instance, in some embodiments, the first treatment and the second treatment are about 3 days, or about 5 days, or about 1 week, or about 2 weeks or about 3 weeks apart.
[00157] In some embodiments, the first and second treatments are about two weeks apart.
[00158] In some embodiments, a single fixed dose of a cell secreting a T cell costimulatory fusion protein [e.g., OX40L-lg) is administered and following the first dose of the cell secreting the T cell costimulatory fusion protein [e.g., OX40L-lg), a second dose is administered along with a fixed dose of a cell secreting a vaccine protein [e.g., gp96-lg) is administered.
[00159] In some embodiments, a single fixed dose of a cell secreting the secreatable vaccine protein [e.g., gp96-lg) is administered with an ascending dose of the cell secreting the T cell costimulatory fusion protein (e.g., OX40L-lg).
[00160] In some embodiments, the methods provided herein can be used for controlling solid tumor growth (e.g., breast, prostate, melanoma, renal, colon, or cervical tumor growth) and/or metastasis. The methods can include administering an effective amount of a cell based therapy as described herein to a subject in need thereof. In some embodiments, the subject is a mammal (e.g., a human).
[00161] The cell based therapies and methods provided herein can be useful for stimulating an immune response against a tumor. Such immune response is useful in treating or alleviating a sign or symptom
associated with the tumor. As used herein, by "treating” is meant reducing, preventing, and/or reversing the symptoms in the individual to which a cell based therapy as described herein has been administered, as compared to the symptoms of an individual not being treated. A practitioner will appreciate that the methods described herein are to be used in concomitance with continuous clinical evaluations by a skilled practitioner (physician or veterinarian) to determine subsequent therapy. Such evaluations will aid and inform in evaluating whether to increase, reduce, or continue a particular treatment dose, mode of administration, etc.
[00162] The methods provided herein can thus be used to treat a tumor, including, for example, a cancer. The methods can be used, for example, to inhibit the growth of a tumor by preventing further tumor growth, by slowing tumor growth, or by causing tumor regression. Thus, the methods can be used, for example, to treat a cancer such as a lung cancer. It will be understood that the subject to which a compound is administered need not suffer from a specific traumatic state. Indeed, the cell based therapy described herein may be administered prophylactically, prior to development of symptoms {e.g., a patient in remission from cancer). The terms "therapeutic” and "therapeutically,” and permutations of these terms, are used to encompass therapeutic, palliative, and prophylactic uses. Thus, as used herein, by "treating or alleviating the symptoms” is meant reducing, preventing, and/or reversing the symptoms of the individual to which a therapeutically effective amount of a composition has been administered, as compared to the symptoms of an individual receiving no such administration.
[00163] As used herein, the terms "effective amount” and "therapeutically effective amount” refer to an amount sufficient to provide the desired therapeutic {e.g., anti-cancer, anti-tumor, or anti-infection) effect in a subject (e.g., a human diagnosed as having cancer or an infection). Anti-tumor and anti-cancer effects include, without limitation, modulation of tumor growth (e.g., tumor growth delay), tumor size, or metastasis, the reduction of toxicity and side effects associated with a particular anti-cancer agent, the amelioration or minimization of the clinical impairment or symptoms of cancer, extending the survival of the subject beyond that which would otherwise be expected in the absence of such treatment, and the prevention of tumor growth in an animal lacking tumor formation prior to administration, i.e., prophylactic administration. In some embodiments, administration of an effective amount of the cell based therapy can increase the activation or proliferation of tumor antigen specific T cells in a subject. For example, the activation or proliferation of tumor antigen specific T cells in the subject can be is increased by at least 10 percent (e.g., at least 25 percent, at least 50 percent, or at least 75 percent) as compared to the level of activation or proliferation of tumor antigen specific T cells in the subject prior to the administration.
[00164] Anti-infection effects include, for example, a reduction in the number of infective agents (e.g., viruses or bacteria). When the clinical condition in the subject to be treated is an infection, administration
of a cell based therapy as provided herein can stimulate the activation or proliferation of pathogenic antigen specific T cells in the subject. For example, administration of the cell based therapy can lead to activation of antigen-specific T cells in the subject to a level great than that achieved by gp96-lg vaccination alone.
[00165] One of skill will appreciate that an effective amount of a cell based therapy may be lowered or increased by fine tuning and/or by administering more than one dose {e.g., by concomitant administration of two different genetically modified tumor cells, or by administering the cell based therapy with another agent {e.g., an antagonist of PD-1 ) to enhance the therapeutic effect (e.g., synergistically). This disclosure therefore provides a method for tailoring the administration/treatment to the particular exigencies specific to a given mammal. Therapeutically effective amounts can be determined by, for example, starting at relatively low amounts and using step-wise increments with concurrent evaluation of beneficial effects. The methods provided herein thus can be used alone or in combination with other well-known tumor therapies, to treat a patient having a tumor. One skilled in the art will readily understand advantageous uses of the cell based therapies and methods provided herein, for example, in prolonging the life expectancy of a cancer patient and/or improving the quality of life of a cancer patient (e.g., a lung cancer patient).
Combination Therapies and Conjugation
[00166] In some embodiments, the invention provides for methods that further comprise administering an additional agent to a subject. In some embodiments, the invention pertains to co-administration and/or co-formulation.
[00167] In some embodiments, administration of a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein, to a patient undergoing a treatment with a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein act synergistically when co-administered with another agent.
[00168] In some embodiments, administration of vaccine protein (e.g., gp96-lg) and one or more costimulatory molecules act synergistically when co-administered with another agent and is administered at doses that are lower than the doses commonly employed when such agents are used as monotherapy.
[00169] In some embodiments, inclusive of, without limitation, cancer applications, the present invention pertains to chemotherapeutic agents as additional agents. Examples of chemotherapeutic agents include, but are not limited to, alkylating agents such as thiotepa and CYTOXAN cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine,
triethylenemelamine, trietylenephosphoramide, triethiylenethiophosphoramide and trimethylolomelamine; acetogenins {e.g., bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; cally statin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins {e.g., cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189 and CB 1-TM1); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gammall and calicheamicin omegall (see, e.g., Agnew, Chem. Inti. Ed. Engl., 33: 183-186 (1994)); dynemicin, including dynemicin A; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, ADRIAMYCIN doxorubicin (including morpholino- doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxy doxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxu ridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as minoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; demecolcine; diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2- ethylhydrazide; procarbazine; PSK polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2, 2', 2"- trichlorotriethylamine; trichothecenes [e.g., T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside
("Ara-C"); cyclophosphamide; thiotepa; taxoids, e.g., TAXOL paclitaxel (Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANE Cremophor-free, albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, 111.), and TAXOTERE doxetaxel (Rhone-Poulenc Rorer, Antony, France); chloranbucil; GEMZAR gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin, oxaliplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; NAVELBINE. vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; irinotecan (Camptosar, CPT-11) (including the treatment regimen of irinotecan with 5-FU and leucovorin); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine; combretastatin; leucovorin (LV); oxaliplatin, including the oxaliplatin treatment regimen (FOLFOX); lapatinib (TYKERB); inhibitors of PKC-a, Raf, H-Ras, EGFR {e.g., erlotinib (Tarceva)) and VEGF-A that reduce cell proliferation and pharmaceutically acceptable salts, acids or derivatives of any of the above. In addition, the methods of treatment can further include the use of radiation. In addition, the methods of treatment can further include the use of photodynamic therapy.
[00170] In some embodiments, inclusive of, without limitation, infectious disease applications, the present invention pertains to anti-infectives as additional agents. In some embodiments, the anti-infective is an anti-viral agent including, but not limited to, Abacavir, Acyclovir, Adefovir, Amprenavir, Atazanavir, Cidofovir, Darunavir, Delavirdine, Didanosine, Docosanol, Efavirenz, Elvitegravir, Emtricitabine, Enfuvirtide, Etravirine, Famciclovir, and Foscarnet. In some embodiments, the anti-infective is an anti bacterial agent including, but not limited to, cephalosporin antibiotics (cephalexin, cefuroxime, cefadroxil, cefazolin, cephalothin, cefaclor, cefamandole, cefoxitin, cefprozil, and ceftobiprole); fluoroquinolone antibiotics (cipro, Levaquin, floxin, tequin, avelox, and norflox); tetracycline antibiotics (tetracycline, minocycline, oxytetracycline, and doxycycline); penicillin antibiotics (amoxicillin, ampicillin, penicillin V, dicloxacillin, carbenicillin, vancomycin, and methicillin); monobactam antibiotics (aztreonam); and carbapenem antibiotics (ertapenem, doripenem, imipenem/cilastatin, and meropenem). In some embodiments, the anti-infectives include anti-malarial agents (e.g., chloroquine, quinine, mefloquine, primaquine, doxycycline, artemether/lumefantrine, atovaquone/proguanil and sulfadoxine/pyrimethamine), metronidazole, tinidazole, ivermectin, pyrantel pamoate, and albendazole.
[00171] Other additional agents are described elsewhere herein, including the blocking antibodies targeted to an immune "checkpoint” molecules.
[00172] In some embodiments, the method comprises administration in combination with an agent that inhibits immunosuppressive molecules produced by tumor cells. In some embodiments, the agent is an antibody against PD-1. In some embodiments, the antibody against PD-1 is selected from nivolumab,
pembrolizumab, pidilizumab, cemiplimab, AGEN2034, AMP-224, AMP-514, and PDR001 .
[00173] In some embodiments, the agent is an antibody against PD-L1. In some embodiments, the antibody against PD-L1 is selected from Atezolizumab (Tecentriq), Avelumab (Bavencio), Durvalumab (Imfinzi), BMS-936559, and CK-301 .
[00174] In some embodiments, the agent is an antibody against CTLA-4. In some embodiments, the antibody against CTLA-4 is selected from ipilimumab, tremelimumab, AGEN1884, and RG2077.
[00175] In some embodiments, the agent is an antibody against 0X40. In some embodiments, the antibody against 0X40 is selected from PF-04518600, BMS-986178, INCAGN01949, MEDI0562, GSK1795091 , and GSK3174998
Subjects and/or Animals
[00176] The methods described herein are intended for use with any subject that may experience the benefits of these methods. Thus, "subjects,” "patients,” and "individuals” (used interchangeably) include humans as well as non-human subjects, particularly domesticated animals.
[00177] In some embodiments, the subject and/or animal is a mammal, e.g., a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, rabbit, sheep, or non-human primate, such as a monkey, chimpanzee, or baboon. In other embodiments, the subject and/or animal is a non-mammal, such, for example, a zebrafish. In some embodiments, the subject and/or animal may comprise fluorescently- tagged cells (with e.g., GFP). In some embodiments, the subject and/or animal is a transgenic animal comprising a fluorescent cell.
[00178] In some embodiments, the subject and/or animal is a human. In some embodiments, the human is a pediatric human. In other embodiments, the human is an adult human. In other embodiments, the human is a geriatric human. In other embodiments, the human may be referred to as a patient.
[00179] In certain embodiments, the human has an age in a range of from about 0 months to about 6 months old, from about 6 to about 12 months old, from about 6 to about 18 months old, from about 18 to about 36 months old, from about 1 to about 5 years old, from about 5 to about 10 years old, from about 10 to about 15 years old, from about 15 to about 20 years old, from about 20 to about 25 years old, from about 25 to about 30 years old, from about 30 to about 35 years old, from about 35 to about 40 years old, from about 40 to about 45 years old, from about 45 to about 50 years old, from about 50 to about 55 years old, from about 55 to about 60 years old, from about 60 to about 65 years old, from about 65 to about 70 years old, from about 70 to about 75 years old, from about 75 to about 80 years old, from about 80 to about 85 years old, from about 85 to about 90 years old, from about 90 to about 95 years old or from about 95 to about 100 years old.
[00180] In other embodiments, the subject is a non-human animal, and therefore the invention pertains to veterinary use. In a specific embodiment, the non-human animal is a household pet. In another specific embodiment, the non-human animal is a livestock animal. In certain embodiments, the subject is a human cancer patient that cannot receive chemotherapy, e.g., the patient is unresponsive to chemotherapy or too ill to have a suitable therapeutic window for chemotherapy {e.g., experiencing too many dose- or regimen-limiting side effects). In certain embodiments, the subject is a human cancer patient having advanced and/or metastatic disease.
[00181] As used herein, an "allogeneic cell” refers to a cell that is not derived from the individual to which the cell is to be administered, that is, has a different genetic constitution than the individual. An allogeneic cell is generally obtained from the same species as the individual to which the cell is to be administered. For example, the allogeneic cell can be a human cell, as disclosed herein, for administering to a human patient such as a cancer patient. As used herein, an "allogeneic tumor cell” refers to a tumor cell that is not derived from the individual to which the allogeneic cell is to be administered. Generally, the allogeneic tumor cell expresses one or more tumor antigens that can stimulate an immune response against a tumor in an individual to which the cell is to be administered. As used herein, an "allogeneic cancer cell,” for example, a lung cancer cell, refers to a cancer cell that is not derived from the individual to which the allogeneic cell is to be administered.
[00182] As used herein, a "genetically modified cell” refers to a cell that has been genetically modified to express an exogenous nucleic acid, for example, by transfection or transduction.
[00183] Technical and scientific terms used herein have the meaning commonly understood by one of skill in the art to which the present invention pertains, unless otherwise defined.
[00184] As used herein, the singular forms "a,” "an” and "the” specifically also encompass the plural forms of the terms to which they refer, unless the content clearly dictates otherwise. As used herein, unless specifically indicated otherwise, the word "or "is used in the "inclusive” sense of "and/or” and not the "exclusive” sense of either/or.” In the specification and the appended claims, the singular forms include plural referents unless the context clearly dictates otherwise.
[00185] The term "about” is used herein to mean approximately, in the region of, roughly, or around. When the term "about” is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. In general, the term "about” is used herein to modify a numerical value above and below the stated value by a variance of 20%. As used in this specification, whether in a transitional phrase or in the body of the claim, the terms "comprise (s)” and "comprising” are to be interpreted as having an open-ended meaning. That is, the terms are to be interpreted synonymously with the phrases "having at least” or "including at least”. When used in the
context of a process, the term "comprising” means that the process includes at least the recited steps, but may include additional steps. When used in the context of a compound or composition, the term "comprising” means that the compound or composition includes at least the recited features or components, but may also include additional features or components. The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.
EXAMPLES
[00186] In order that the invention disclosed herein may be more efficiently understood, examples are provided below. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting the invention in any manner.
Example 1: Planned dose range finding studies in mice to support clinical dosing
[00187] The following nonclinical study is conducted in mice using a surrogate (specific for animal species to be tested) cell-based vaccine for HS-130, which is a genetically engineered human lung adenocarcinoma cell line secreting OX40L-lg fusion protein, also (referred to as mHS-130 (B16F10) and HS-1 10 (referred to as mHS-1 10 (B16F10), generated from a B16F10-ova cell line, to determine a dose of mHS-130 when co-administered with a fixed dose of mHS-110. The starting dose of HS-130 in humans is based on the dose of mHS-130 that generates a minimum response (MABEL) in the mouse model based on the proportional use of mHS-110 in mice vs. the current dose of HS-110 in humans.
[00188] In previous studies, the dose of murine HS-110 (mHS-1 10) in mice was determined to be 1 x 106 cells. It was further determined by quantitative ELISA that this number of cells secrete 290 ng of gp96-lg fusion protein per 24 hours. In order to determine the ratio of gp96-lg to OX40L-lg in this system, a murine form of HS-130 [mHS-130 (B16F10)] secreting the species specific OX40L-lg protein is generated. It has also been determined by quantitative ELISA that mHS-130 (B16F10) secretes 850 ng of OX40L-lg per 106 cells per 24 hours.
Experimental Design
[00189] Wild-type C57BL/6 mice are adoptively transferred with 106 OT-I* and 106 OT-llFoxP3-rfp congenic cells. These cells are transgenic CD8+ and CD4+ T cells designed to recognize a specific ovalbumin peptide. When transferred into wild-type mice these cells join the other murine lymphocytes in circulation until they encounter their cognate antigen and can be activated. Both of the murine vaccine cell lines mHS-1 10 (B16F10) and mHS-130 (B16F10) are engineered to produce ovalbumin, a protein not naturally expressed in the mouse. Two days after adoptive transfer the mice are vaccinated with mHS-1 10 (B16F10) and mHS-130 (B16F10) according to the doses described in the study protocol below
(Table 1). Mice are followed for 14 days. Periodically, peripheral blood is collected from the tail vein and analyzed for the frequency of CD8+ and CD4+ T cells by flow cytometry. The development of CD4+ FoxP3+ T regulatory subset of cells are further characterized in this system by the induction of red fluorescent protein upon expression of the master regulator FoxP3. This helps with studying the influence of vaccination platform on antigen specific T lymphocytes over time, that have been shown to be critical for effective tumor reduction and control. Additionally, the weight of each animal is tracked at the time of blood draw, as an indicator of overall health throughout the study.
Table 1 : Study protocol for dose-range finding study in mice
[00190] In-vitro assay comparing signaling strength of murine and human OX40L-lg in a human 0X40 receptor signaling assay:
[00191] To characterize the signaling activity of OX40L derived from mouse and human species on the human 0X40 receptor transfected in the human Jurkat cell line to determine the equivalence of cross species signaling. A side by side comparison of muOX40L-mlgG1 (mouse derived OX40L), and huOX40L-hlgG4 (human derived OX40L) of NFKB signal induction through human 0X40 receptor binding was done using a Jurkat/OX40 cell based system. Cells were stimulated with muOX40L-mlgG1 or huOX40L-hlgG4 with the maximum concentration of 1 g/ml. Each ligand was tested in duplicate wells on the same plate. The luciferase activation was measured after 5 hours using Bio-Glo reagent from Promega and the relative luminescence was measured using a luminometer. The ECso values were calculated using a four-parameter logistic curve analysis. The calculated ECso values indicate that the activity of the mouse and human derived OX40L are very similar against the human 0X40 receptor. The
ECso results from this experiment, as shown in FIG. 2, align with the observation of high sequence
homology between human and mouse OX40L and suggest that the mouse is a good predictor of clinical pharmacokinetics.
Example 2: Phase 1 Clinical Trial, Dosage Form, Route of Administration and Dosing Regimen
[00192] The drug product is a viable whole cell vaccine that has been irradiated to render cell replication-incompetent while expressing the co-stimulatory fusion protein OX40L-lg, which is the ligand for the 0X40 receptor, a member of the TNF-receptor superfamily. The drug product is a viable whole cell vaccine derived from a human lung adenocarcinoma cell line. The cell line is transfected with a 7192 bp plasmid cDNA‘pcDNA3.4 OX40L-lg' stably expressing the cDNA for OX40L-lg to develop an irradiated whole cell vaccine as shown in FIG. 1. HS-110 refers to (viagenpumatucel-L); genetically engineered human lung adenocarcinoma cell line secreting gp96-lg fusion protein, and HS-130 refers to genetically engineered human lung adenocarcinoma cell line AD100, secreting OX40L-lg fusion protein.
[00193] A Phase I, first-in-human, dose-escalation study to evaluate the safety and immunologic dose of HS-130 alone and in combination with HS-110 in patients with solid tumors refractory to standard care is undertaken.
[00194] Primary Objective: To study safety and tolerability of HS-130 alone and in combination with HS-110 in patients with solid tumors refractory to Standard of Care (SOC).
[00195] Secondary Objective: To determine the immunologic dose of HS-130 with a fixed dose of HS- 110 administered in combination in patients with solid tumors refractory to SOC. To study the immunological effect generated by HS-130 in combination with HS-110 by determining total peripheral blood mononuclear cell (PBMC) counts and activation state of PBMC subsets using flow cytometry.
[00196] Exploratory Objective
[00197] 1) To evaluate archival tissue for shared antigen expression with HS-110 by RNA-seq; 2) To evaluate immune reactivation response via ELISPOT using IFNy, granzyme B (gzB) production as functional readouts; 3) To determine the presence of a specific cytokine signature in response to combination treatment with HS-130 and HS-110; and 4) To determine clinical response to combination treatment with HS-130 and HS-110.
[00198] Methodology:
[00199] This is a first-in-human, Phase I trial of HS-130 alone and in combination with HS-110 in a mixed population of patients with advanced solid tumors refractory to SOC. Both HS-130 and HS-110 are genetically modified, viable, replication-incompetent cancer cells, designed to stimulate an immune response when administered into the intradermal layers of the skin. The purpose of the study is to study the safety and immunological response associated with the treatment. In the study, patients who meet
the inclusion/exclusion criteria receive escalating doses of the HS-130 cells using a 3 + 3 design. The first cohort start treatment with a single fixed dose of HS-130 based on the minimum anticipated biological effect level (MABEL) established in animal models. After a safety assessment interval of 2 weeks following the first dose of HS-130, the patient is administered the same dose of HS-130 along with a fixed dose of HS-110 cells (1 x 107 cells, which is an established safe dose in the ongoing Phase 2 study of HS-110). In the absence of any safety issues 2 weeks after the combination treatment, the patient can continue the combination treatment of HS-110 + HS-130 administered bi-weekly for 6 months or until disease progression, death, patient withdrawal of consent, investigator decision to remove patient, or intolerable toxicity, whichever occurs first.
[00200] Three patients are enrolled in each dose cohort. After the first patient, the second and third patients has a 1 week staggered delay following the first patient dose. Once all 3 patients have completed the first cycle of dosing (/.e. 4 weeks, with one dose of HS-130 on Day 1 , and one combined dose on Day 15), a review of the safety and tolerability of the treatment administered among the patient cohort is reviewed by an Investigator/Sponsor dose escalation committee. If no Dose Limiting Toxicity (DLT) occurs, then the dose escalation committee may recommend enrollment at the next higher dose level. If DLT occurs in one patient, then up to 3 additional patients is enrolled and treated at the dose level. If less than or equal to one in six patients experiences a DLT, then the dose escalation committee recommends enrollment at the next higher dose level. The conduct and completion of each subsequent dose cohort follows in similar fashion until DLT occurs in 2 patients among a total of up to 6 treated patients at a dose level.
Schematic for treatment regimen for patients in the study:
[00201] Should an immunologically active dose be identified before reaching MTD, then the sponsor may decide to discontinue further dose escalation. Any patient who does not complete the first cycle of
treatment (i.e. at least one combination dose) is replaced. It is predicted that 4 dose levels of HS-130 are explored, and 12 to 24 patients are enrolled on the trial. All patients are monitored for extensive safety assessment including serum cytokines/chemokines, and immune phenotype profiling of immune cell subsets by flow cytometry. The immune response is evaluated by ELISPOT using HS-110 lysate and HS-110 specific peptides for IFNy and Granzyme B production. Safety is assessed by frequency of adverse events (AEs), evaluation of clinical laboratory parameters (hematology, and biochemistry), weight, vital signs, electrocardiogram (ECG), performance status, physical exams (PEs), and recording of concurrent illness/therapy and adverse events. CTCAE version 5 is used to grade all toxicities.
[00202] Number of Patients: Up to 12-24 total patients are enrolled.
[00203] Inclusion Criteria: Patients must meet all of the following inclusion criteria before they are allowed to participate in the trial: Patients with select solid tumor types (defined as those having CTA overexpression overlap with at least 10 CTAs overexpressed by HS-110) who have failed available standard therapy or who are not candidates for standard therapy, and for whom, in the opinion of the investigator, experimental therapy with HS-110 + HS-130 may be beneficial. Age ³ 18 years. Have an acceptable organ function. Have an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1. Life expectancy of at least three months. Have fresh or archival tumor tissue available at screening. Patients, both females and males, of childbearing/reproductive potential must agree to use adequate contraception while included in the trial and for six months after the last treatment with HS-130 and/or HS-110. Patients must sign the informed consent form.
[00204] Exclusion Criteria: If any of the following apply, the patient must not enter the trial: Clinically significant cardiac disease, congestive heart failure and/or uncontrolled hypertension. Known or clinically suspected leptomeningeal disease. Stable, previously treated metastases in the brain or spinal cord, are allowed as long as these are considered stable (by CT or MR), and not requiring systemic corticosteroids. History of ³ grade 3 allergic reactions as well as known or suspected allergy or intolerance to any agent given in the course of this trial, live cell therapies, or live vaccines. History of suspected cytokine release syndrome (CRS). Known immunodeficiency disorders. Ongoing or current autoimmune disease (history of checkpoint inhibitor immune related adverse events are permissible if resolved). Any condition requiring concurrent systemic immunosuppressive therapy. Major surgery within four weeks before first IMP administration. Any ongoing anticancer therapy including; small molecules, immunotherapy, chemotherapy, monoclonal antibodies or any other experimental drug. Prior therapy must be stopped within four weeks before first infusion in the study, or 5 half-lives, or twice the duration of the biological effect of the investigational product (whichever is shortest). Adjuvant anti-hormonal treatment(s) for prior breast cancer or prostate cancer are allowed. Known current malignancy other than inclusion diagnosis.
Prior curable cancer with complete remission for >2 years is allowed. Any other ongoing significant, uncontrolled medical condition. Received a live vaccine within 30 days prior to first dose of study drug. Clinically significant active viral, bacterial or fungal infection requiring: Intravenous treatment with antimicrobial therapy completed less than two weeks prior to first dose, or Oral treatment with antimicrobial therapy completed less than one week prior to first dose. Prophylactic treatment (e.g., for dental extractions) is allowed. Known positive serology for human immunodeficiency virus (HIV), hepatitis B, or hepatitis C (except in cases of immunity after cured infection). Substance abuse, medical, psychological or social conditions that may interfere with the patient's participation in the trial or evaluation of the trial result. Women who are pregnant or breast feeding. Dose and Mode of Administration: Each patient is administered an intradermal injection of HS-130 on Day 1 and another intradermal injection of HS-130 at the same dose with a fixed dose of HS-110 (1 x 107 cells) on Day 15. Patients not progressing may continue treatment at the discretion of the investigator.
[00205] Dose Limiting Toxicity: DLT is defined as any non-acceptable (as defined below) treatment related toxicity (i.e., not attributable to the active disease, disease-related processes under investigation or intercurrent illness) observed during the first 28 days of study treatment. Ongoing safety events beyond Cycle 1 is reviewed across all cohorts during the study to help inform dose escalation decisions.
[00206] DLT includes: Hematological toxicities ³ CTCAEv5 grade 3. Non-hematological toxicity ³ CTCAEv5 grade 3. Any other toxicity (greater than at baseline), considered clinically significant and/or unacceptable, and that does not respond to supportive care and results in a disruption of the dosing schedule of more than 14 days.
[00207] DLT excludes: Grade 3 self-limited or medically controllable toxicities {e.g., fever without ³ Grade 3 neutropenia, nausea, vomiting, diarrhea, fatigue). Electrolyte disturbances that are managed to grade 1 or less with supplemental therapy.
[00208] Data Monitoring Committee: A Data Monitoring Committee (DMC) evaluates the data obtained at each dose level and recommends whether the dose should be escalated as per protocol, revised to a lower level or intermediate level, halted altogether or more patients are required at the same dose level to evaluate safety. Following each DMC meeting, a sponsor safety committee meeting is held to discuss and confirm actions recommended by the DMC.
[00209] Duration of Treatment: Upon completion of the first treatment cycle (i.e. 4 weeks, with one dose of HS-130 on Day 1 , and one combined dose with HS-110 on Day 15), in the absence of disease progression or unacceptable toxicity, patients may continue to be treated with the combination of HS-130 + HS-110 at the same dose bi-weekly for 6-months or until disease progression, death, patient withdrawal of consent, investigator decision to remove patient, or intolerable toxicity, whichever occurs first.
[00210] Criteria for Evaluation:
Primary Endpoints
• Safety and Tolerability: Measured by the frequency of treatment emergent adverse events (TEAEs)/serious adverse events (SAEs), including clinically significant (CS) abnormal laboratory parameters, ECGs, PEs, and vital signs in patients receiving at least 1 dose of study drug.
• Maximum Tolerated Dose (MTD; highest dose level at which less than one-third of at least 6 patients experienced a DLT during the first treatment cycle), OR definition of an immunologically active dose that makes further dose escalation redundant (i.e. plateau of immune markers or trigger of immunosuppression).
Secondary Endpoints
• Peripheral blood IR analysis of surface markers, CD3, CD4, CD8, CD19, CD25, CD39, CD45, CD56, CD73, FoxP3, Ki-67, and ICAM-1 as surrogates for T-cell activation and proliferation.
Exploratory Endpoints
• Bioinformatic analysis of RNA-seq data generated with comparison to historical gene expression data.
• Calculation of responding cell numbers over the course of treatment after subtraction of appropriate patient controls.
• Analysis by Luminex multiplex panel to detect changes in pro-inflammatory serum cytokines before and after treatment with HS130 and HS110.
• Patient disease status is monitored by clinical and radiological assessment as per the institutional standard. Patient clinical response is evaluated as complete response, partial response, stable disease, or progressive disease per Investigator assessment.
[00211] Safety: All patients who have received any component of study treatment are considered evaluable for safety. Safety is assessed by means of physical examination, weight, vital signs, performance status, laboratory evaluations (hematology, biochemistry including cytokines and C-reactive protein), electrocardiogram (ECG), and recording of concurrent illness/therapy and adverse events. CTCAE version 5 are used to grade all toxicities. All related adverse events are monitored until resolution. Patients are monitored for safety and concomitant medications throughout the study. Cytokines {e.g., IFNy, IL-1 b, IL-2, IL-4, IL-6, IL-10, IL-12 p70, IL-17A, TNFa) are be monitored once after each dose and repeatedly in relation to clinically observed reactions after drug injections.
[00212] Statistical Methods: All analyses are descriptive. Categorical variables are presented with numbers and, if meaningful, percentages. Continuous variables are presented by n, mean, median, standard deviation and range (min and max) as appropriate. Presentations are by each dose cohort.
Efficacy: This is an exploratory trial and therefore no sample size calculations have been performed. Safety Population: All patients who receive at least 1 dose of study drug are evaluated for safety.
Example 3: Mouse HS-110 (B16F10-OVA-gp96) and mouse HS-130 (B16F10-OVA- OX40L) immunization at a“narrow” dose-range to correlate CD8+ T-cell expansion to tumor growth
[00213] The experiments of this example demonstrate a best ratio of a mouse HS-110 (mHS-110; B16F10-OVA-gp96-lg) to a mouse HS-130 (mHS-130; B16F10-OVA-OX40L) for the generation of a primary and secondary tumor-specific CD8+ T-cell response, and demonstrate how anti-tumor T-cell expansion correlates to tumor growth control, in vivo.
[00214] As disclosed in the prior examples, HS-110 is a lung adenocarcinoma cancer cell line. HS- 1 10 secretes gp96-lg, which presents antigens to prime and expand CD8+ T cell responses. Preclinical studies have suggested that the addition of secreted T cell costimulatory molecules, OX40L-lg in combination with Gp96-lg in a cellular system expressing a defined antigen, enhances T cell immunity and results in elimination of tumors (Fromm G et al., Cancer Immunol Res. 2016 Sep 2;4(9):766-78.). Therefore, a mouse cell line was devised that secretes only OX40L-lg, herein referred to as "mouse HS- 130” or "mHS-130”, which was used in combination with mHS-110 (See Example 1, above). The best ratio of mHS-110 to mHS-130 for the generation of a primary and secondary CD8+ or CD4+ T cell pool was investigated in the following experiments. The experiments of this example further included a tumor challenge arm to better understand how in vivo expansion of CD8+ T cells correlates to anti-tumor immune responses and tumor growth control.
[00215] Experimental Design and Methodology
[00216] An image showing the full study design can be found in FIG. 3.
[00217] OT-1 purification, adoptive T-cell transfer, mHS-110/mHS-130 dosing, and flow cytometry staining: T-cell receptor (TCR) transgenic mouse CD8+ (OT-I) cells were isolated from OT-I-GFP bred mice using Easy Sep Mouse CD8 T Cell isolation kit (cat # 19853A) and injected into each C57BL/6 mouse intravenously (i.v) through lateral tail vein with 1 million OT-I cells suspended in HBSS (GIBCO 14175-095). Two days after injecting OT-I, all the mice were tail bled for baseline and 4 hours later, mHS-1 10 (B16F10-OVA-gp96-lg) cells and mHS-130 (B16F10-OVA-OX40L-lg) were treated with 10 pg/mL of Mitomycin-C (Sigma-Aldrich cat# M0503) for 3 hours and given intraperitoneally (i.p.) to each group accordingly. Mice were divided into 7 groups with 5 mice per group, except for the parental group which had 3 mice. Animals were dosed based on nanogram expression level (per 106 cells per 24hr) of gp96-lg or OX40L-lg per the design shown in FIG. 3 and Table 2 below.
[00218] Table 2: Animal Dosing Design
[00219] Mice were tail bled consecutively on days 3, 5, 7, 10, and 14 post-immunization, and days 17, 19, 21 , 24, 28, 33, 38 and 41 post-boosts into heparinized PBS (10 units/ml), and lysed using ACK lysis buffer (150 mM NhUCI, 100 mM KHCO3 and 10 mM EDTA 0.2 Na, pH 7.2) for 3 minutes, and neutralized with 1 X PBS. Samples from the OT-I transfer experiments were then centrifuged at 300 g for 5 minutes, supernatant was removed, and the cell pellet stained with anti-CD3 (20 pg/mL), anti-CD4 (40 pg/mL) and anti-CD8 (5 pg/mL) antibody cocktail made in FACS buffer using Alexa Fluor 700 anti-mouse CD3 (Bio legend cat # 100216), Alexa Fluor 647 anti-mouse CD4 antibody (Biolegend Cat # 100530) and Brilliant Violet 421 anti-mouse CD8alpha antibody (Biolegend Cat # 100738) for 30 minutes at 4°C.
[00220] Intracellular Cytokine Staining and Flow Cytometry: Splenocytes (1 *106) were incubated with synthetic peptides; SIINFEKL (SEQ ID NO:41), gp100, TRP-1 , TRP-1 -Variant, or TRP-2 in the wells of a 96-well plate at 37°C and 5% C02. Synthetic peptides were added to a final concentration of 0.5 mM with Golgi stop and incubated for 4-10 hours depending on the peptide. Plates were spun, medium was removed, and cells were resuspended with surface markers CD8 and CD3 and incubated at 4°C for 20 minutes. Cells were washed, resuspended in 50 pi of BD Cytofix/Cytoperm, and incubated at 4°C for 20 min before another two washes and staining with anti-IFN-g. Cells were washed once before acquisition and analysis of fluorescence. Analysis was done using FlowJo software (Tree Star Inc.); events were gated for live lymphocytes on FSC x SSC followed by CD8+ cells using CD8 x CD3 and displayed as CD8 x IFN-y.
[00221] B16F10-OVA tumor challenge and volume calculations: Melanoma B16F10 cells were harvested and resuspended at a concentration of 5x105 cells/100 mI in a volume of 80 mI HBSS and 20 mI Matrigel. C57BL/6 mice were subcutaneously injected with 100 mI of B16F10 cells (5x105 cells/mouse) on the inner abdomen 29 days post-OT-1 transfer and 28 days post-primary vaccination, designated "day 28”, as shown in study design (FIG. 3). The tumor size was measured and documented every 3 days with a caliper, starting on day 7, and calculated using the formula (AxB; A as the largest and B as the smallest diameter of tumor). Tumor growth was documented as standard error mean. To record the
survival of the tumor-bearing mice, either natural death or a tumor volume greater than 450 mm2 leading to death was counted as death. Each experimental group included five animals.
[00222] Tumor tissue digestion for Tumor Infiltrating Lymphocytes (TILs): The MACS Miltenyl Biotec tumor dissociation kit was used for this procedure (cat# 130096-730).
[00223] ELISPOT Assay: Splenocytes were harvested and red blood cell lysis buffer (cat#36858500, Roche) was used to eliminate red blood cells. Cells were washed in IMDM medium and pelleted. Counted cells were resuspended in IMDM with 10% FBS. Each ELISPOT well received 1 million cells, in a total volume of 200 pi. Treatments included the use of B16F10-OVA parental line lysates (vial of 10 million cells; freeze-thawed three times) at a 10-fold dilution, immunodominant epitopes for B16F10 tumors: "gp100/pmel” (EGSRNQDWL (SEQ ID NO:38)), "TRP-1/gp75” (TWHRYHLL (SEQ ID NO:39)), TRP-2 (SVYDFFVWL (SEQ ID NO:40)) and the MHC-I restricted peptide for H2b haplotype for OVA (SIINFEKL (SEQ ID NO:41)). All peptides were used at a final concentration of 10 ug/mL. PMA/lonomycin was used as a positive control at "1 to 400 dilution” 100 nM of PMA, 1.6 mM ionomycin (500x stock from eBioscience, cat#00-4970-03, and whole chicken ovalbumin protein "OVA” at 250 pg/ml.
[00224] Interferon-gamma capture ELISPOT assay was carried out as previously published (Klinman DM et at., Current Protocols in Immunology. 1994 June (1): 6.19.1-6.19.8). Briefly, MAIP N45 Milipore 96-well filtration plates were coated with anti-mouse IFN-g purified monoclonal antibody (clone AN18) in PBS saline at 10 pg/mL, 50 mI/well, at 4°C, overnight. The next day, cells were washed in IMEM medium plus 10% FBS at 200 ul/well, washed three times, then blocked for 1 hour at 25°C. Cells were processed as described above and 1 million or 100,000 splenocytes were added to each well and treated per the description above with various peptide stimulations. Each plate was incubated in a 37°C, 5% C02, non vibrating incubator for 16 hrs or overnight. At the end of the incubation, cells were flicked into the sink to stop the culture and washed with PBS + 0.1% Tween-20 three times and blotted hard onto paper towels to get rid of residual liquid after the last wash. Secondary biotinylated antibody was added at 50 pi per well, 4 pg/mL of anti-IFN-g (clone AN18), incubated at 25°C for 2 hrs. Plates were then washed again with PBS+0.1 % Tween-20 three times. Peroxidase-conjugated streptavidin from Jackson research labs was then added at a 1 to 1000 dilution from stock concentration in PBS + 0.1 % Tween-20 + 2% BSA, 50 pi per well, 30-minute incubation at 4°C. Plates were then washed with PBS + 0.1 % Tween three times. The plastic backing of the ELISPOT plate was removed and the entire plate was submerged and soaked in PBS + 0.1% Tween-20 for 1 hour at 25°C. The plate was then washed in PBS to remove the tween, then 100 pi of the AEC substrate (Vector kit) added in 100 mM of TRIS buffer at pH 8.2 was added to each well to allow for development of spots. At the end of the incubation, 10-20 minutes, all plates incubated for the same time, the reaction as stopped by rinsing the plates with tap water and air dried.
[00225] Plates were scanned and counted in an AID Autoimmun Diagnostika GMBH iSpot monochromatic ELISPOT reader (2018). Counting parameters for each plate remained consistent across all plates that were developed together as to prevent bias, as spot development, density and magnitude is influence by the operator, reagent lots and final substrate development time.
[00226] Experimental Results
[00227] Na'ive OT-I GFP+ CD8+ T cells expressing \/a2L/b5 and specific for H-2Kb/OVA25/-264 were adoptively transferred into na'ive WT C57BL/6 mice. Recipient mice were injected with mHS-110 at a fixed dose of 1 million cells (290 ng of gp96-lg) with different ratios of mHS-130, in which a 1 to 1 ratio of mHS-1 10 to mHS-130 was equivalent to 290 ng of gp96-lg to 290 ng of OX40L.
[00228] After vaccination, OT-I GFP+ CD8+ T cells were analyzed in the blood on days 0, 3, 5, 7, 10, and 14, representing the acute expansion phase and the contraction phase of CD8+ T cell responses. Based on accumulation of the transferred cells, the generation of the primary effector pool was found to peak on day 7 (FIG. 4, FIG. 6A, FIG. 6B) and contract by day 14 post-vaccination (FIG. 6A, FIG. 6B). Significant increases were found on day 7 in the 1 to 1 , the 1 to 3 and the 1 to 10 ratios of mHS-110 to mHS-130 (FIG. 6A, FIG. 6B) (*p<0.05; **p<0.01). Furthermore, the 1 to 1 ratio had significant increases on days 10 and 14 post-vaccination (*p<0.05).
[00229] Following this analysis, a boost vaccination was given on day 14, and OT-I GFP+ CD8+ T cells were analyzed in the blood on days 17, 19, 21 , 24, and 28, representing the secondary expansion phase and the contraction phase of CD8+ T cell responses (FIG. 5, FIG. 6A, FIG. 6B). Based on accumulation of the transferred cells, the generation of the memory effector pool was found to peak and increase in percentages by day 21 post-challenge and contract by day 28 post-challenge. Significant increases were found on day 21 , 24 and 28 in the 1 to 1 ratio of mHS-110 to mHS-130 (FIG. 6A, FIG. 6B) (*p<0.05).
[00230] With each subsequent boost of mHS-110 with mHS-130, the expansion of antigen-specific T- cells became less. Tumor was given s.c., at 500,000 B16F10-OVA cells per mouse on day 28, with another boost of mHS-110 with mHS-130 given on day 31 . This resulted in a non-significant increase in antigen-specific CD8+ T-cells (FIG. 6A, FIG. 6B), however without outliers removed, the 1 to 1 ratio provided the best and most consistent response to vaccine re-challenge (FIG. 6B).
[00231] Looking at the endogenous response, at day 54, the end-of-study, the percent of CD8+ T-cells is elevated in only those ratio groups that showed the greatest anti-tumor response, 1 to 1 and 1 to 10 (FIG. 7A), *p<0.05 as compared to mHS-110 alone. Splenocytes rechallenged, with B16F10, H2Kb restricted, immunodominant peptide, gp100, directly ex vivo, generated IFN-g release by tumor-specific CD8+ T-cells, however, all groups except the 1 to 0.1 group showed no difference compared to mHS- 110 alone, suggesting that such antigen specific T-cells most likely migrated directly to the tumor, which
resulted in the observed growth retardation (FIG. 7B, FIG. 7C). This is supported by data showing an increase in CD8+ TILs in remaining tumors (FIG. 8A). Other peptides were also tested including TRP-2, TRP-1 , and TRP-1 -variant. The H2Kb restricted peptide of OVA, SIINFEKL (SEQ ID NO:41), was also tested, however, OT-1 frequencies were very low at the day 54 timepoint, that only a weak response was noted (data not shown).
[00232] Endogenous immune responses to vaccination and tumor burden was examined using end- of-study ELISPOTs from splenocytes. In agreement with the reported flow cytometry results and tumor growth, responses to tumor lysate, various B16F10 immunodominant peptides and OVA all followed the predicted ratios and appeared to be dependent on the presence of mHS-130 (FIG. 7D). Responses to various stimulants were significant compared to mHS-110 alone by the Mann-Whitney non-parametric statistical test.
[00233] With regard to flow cytometry plots, endogenous CD4+ T-cells expanded in frequency for the 1 to 1 and 1 to 10 ratio dose groups, suggesting the need for Th1 help to generate anti-tumor cytotoxic T-lymphocytes, in vivo (FIG. 7E). Effector (CD62U0 CD44hi) and na'ive (CD62Lhi CD44'°) CD4+ and CD8+ endogenous T-cell frequencies were also measured; and no biologically significant response was found between the groups (data not shown). Also, no biologically significant response was seen for PD-1 + CD8+ endogenous T-cells nor ILRG+/- IL-7R+/- memory CD8+ T-cell subsets, for any group tested (data not shown).
[00234] Tumor infiltrating lymphocytes (TILs) were quantitated for each remaining tumor mass, per animal, at the end of the study, day 54. The proportion of CD8+ TILs in the 1 to 1 and 1 to 10 groups were 51-fold and 118-fold, respectively, over that of the 1 to 0.1 vaccination ratio of mHS-110 to mFIS- 130 (FIG. 8A, FIG. 8B). The 1 to 1 group had two animals that failed to establish stable tumors (full tumor growth inhibition), and the 1 to 10 group had three animals with full tumor growth inhibition. For this reason, proper statistics for TIL percentages cannot be performed with less than three samples, thus the 1 to 10 group is ineligible for statistical analysis. Flowever, comparing the 1 to 1 group to mHS-110 alone (FIG. 8A, FIG. 8B; *p<0.05), it was observed that significantly more CD8+ TILs were found in these tumors, and this was true when compared to the 1 to 0.1 group as well. In both cases, there was a dose- dependent increase in the proportion of CD8+ TILs with vaccination ratio with the 1 to 1 and 1 to 10 ratio giving the best response.
[00235] Tumor volume was measured over time, from the point of tumor cell inoculation until end of study, a total of 25 days of logarithmic growth. Since this was a protection, prophylaxis, study, tumor delay was measured, rather a therapeutic response to established tumors. As shown in FIG. 10A, only two ratios, 1 to 1 (290 ng of gp96-lg to 290 ng of OX40L-lg) and 1 to 10 (290 ng of gp96-lg to 2900 ng of
OX40L-lg) of mHS-110 to mHS-130, gave a consistent and significant tumor growth inhibition as compared to the mHS-110 group alone, with greatest separation observed for days 21 -25 (**p<0.01). Individual tumor growth curves are shown in FIG. 10B to show individual animal variance. Measuring end-of-study tumor mass confirmed what tumor volume had demonstrated, in that both the 1 to 1 and 1 to 10 ratio gave the best response, as compared to mHS-110 alone (FIG. 9). For tumor volume, the 1 to 1 , and 1 to 10 ratio were not significantly different.
[00236] Tumor growth inhibition, ex vivo anti-tumor T-cell responses and TIL infiltration correlated with reduced PD-1 + expression on CD8+ T-cells, observed in the 1 to 1 dose ratio group (FIG. 12A, FIG. 12B; *p<0.05). Although not significantly different from mHS-1 10 alone group, the 1 to 10 ratio also showed a trend decrease in PD-1 expression. Expression of PD-1 occurs during initial T-cell activation, but under constant re-stimulation or in the presence of cognate antigen, PD-1 is expressed by exhausted T-cells (Simon S. et at., Oncoimmunology. 2018 September 7 (1 ): e1364828). Elevated expression of PD-1 was seen in those groups with the greatest tumor burden (1 to 0.1 , mHS-1 10 alone and parental B16F10- OVA alone), which suggested that cells have become exhausted from continued anti-tumor fighting, but those that have received proper stimulation with OX40L, via mHS-130 vaccination, show lower PD-1 expression and less tumor burden. This may explain why the 1 to 1 and 1 to 10 ratio dose groups showed the lowest PD-1 expression in the spleen. Expression of PD-1 was also measured in the tumor, by looking at TILs, but the frequency was below the limit of quantitation making analysis difficult (data not shown).
[00237] Day 54 spleen OT-1 frequencies and absolute counts were measured for remaining groups at the end of the study. Only the 1 to 1 ratio produced long-lived circulating OT-1 cells above all other groups tested (FIG. 11 A, FIG. 11 B; *p<0.05). This data strongly suggested that the 1 to 1 ratio of gp96- Ig to OX40L-lg produces the best anti-tumor T-cell expansion response for both immediate and long-term immune responses, and that this directly correlates to tumor growth inhibition, in vivo.
[00238] Example 4: Study ofgp96-lg (mHS-110) to OX40L-lg (mHS-130) dose ratios
[00239] In this example, experiments were conducted to determine best gp96-lg (mHS-110) to OX40L- Ig (mHS-130) dose ratios that result in CD8+ T cell expanstion and tumor growth inhibition. HS-1 10 is a lung adenocarcinoma cancer cell line that secretes gp96-lg, which presents antigens to prime and expand CD8+ T cell responses. In the present example, a mouse cell line (mHS-130) was developed that secretes only OX40L-lg.
[00240] In the present study, mHS-130 (secreting gp96-lg fusion protein) was used in combination with mHS-1 10 (secreting OX40L-lg fusion protein). An objective of the study was to determine ratio of mHS- 1 10 to mHS-130 most suitable for generation of a primary and secondary CD8+ or CD4+ T cell pool.
Thus, this study tested in tumor-bearing animals a variable ratio, in a dose-escalation manner, to determine the ratio(s) and dose(s) that result in the most effective CD8+ T-cell expansion and tumor growth inhibition combination.
[00241] Experimental Design and Methodology
[00242] FIG. 13 illustrates a design of the present study.
[00243] OT-1 purification, adoptive T-cell transfer, mHS-110/mHS-130 dosing, and flow cytometry staining
[00244] T-cell receptor (TCR) transgenic mouse CD8+ (OT-I) cells were isolated from in-house bred OT-I-GFP mice using Easy Sep Mouse CD8+ T Cell isolation kit (cat # 19853A) and injected into each C57BL/6 mouse intravenously (i.v.) through lateral tail vein with 1 million OT-I cells suspended in HBSS (GIBCO 14175-095). Two days after injecting OT-I, all the mice were tail bled for baseline and 4 hours later, mHS-110 (B16F10-OVA-gp96-lg) cells and mHS-130 (B16F10-OVA-OX40L-lg) were treated with 10 pg/mL of Mitomycin-C (Sigma-Aldrich cat# M0503) for 3 hours and given intraperitoneally (i.p.) to each group accordingly. Mice were divided into 10 groups with 5 mice per group. Three different treatment ratios were provided with escalating doses within each ratio group and all compared against mHS-1 10 (gp96-lg) alone, the control group (ratio 1 to 0). Animals were dosed based on nanogram expression level (measured as ng/106 cells/24 hrs) of gp96-lg or OX40L-lg, as shown in Tables 3 and 4 below. As shown in the study design FIG. 13, dose ratios of gp96-lg to OX40L-lg were 1 :1.3, 1 :2.5, 1 :5, and 1 :0 (mHS-110 (gp96-lg) alone), and each dose was tested at three different dose levels ("low,” "medium,” and "high”). Mice were boosted on days 14 and 31 with the same ratios of mHS-110 and mHS-130 as in the primary phase, and OT-I GFP+ CD8+ T cells were analyzed in the blood days post-challenge. T umor was provided on day 28. Consecutive bleeds were collected from the peripheral blood and analysis was performed by flow cytometry on both exogenous, adoptively transferred, OT-1 and endogenous CD8+ and CD4+ T-cells for activation, and short (SLECs) and long-term (MPECs) memory markers as outlined in the methods section. Tumor growth kinetics, response rates and infiltrating lymphocytes, were also quantitated.
[00245] Cell line protein expression data for mHS-110 and mHS-130
[00246] The amount of murine gp96 protein expressed by the mHS-110 cells was determined by ELISA. For each sample to be tested, one million B16F10-Ova9 parental and mHS-110 cells were plated in a 6-well tissue culture plate in a total volume of 1 ml each. Cells were incubated at 37°C with 5% CO2 for 24 hours at which point the supernatants were harvested. Supernatants were then centrifuged at 2500 rpm for 5 minutes to pellet any cell debris. Clarified supernatants were then transferred to new 1.5 ml tubes and stored at -80°C. Each sample tested was from a fresh vial of mHS-1 10 cells thawed and
expanded.
[00247] To perform the ELISA, 96-well plates (Corning, cat# 9018) were coated with 2 ug/ml of sheep anti-gp96 (R&D Systems, cat# AF7606) in carbonate-bicarbonate buffer. Plates were sealed and stored at 4°C overnight. Plates were then washed 4 times with 1X TBST (VWR, cat# K873) and then blocked with 1X casein solution (Sigma-Aldrich, cat# B6429) for 1 hour at room temperature. The plates were then washed 4 times with 1X TBST and a human gp96-mouse Fc standard (Thermo Fisher Scientific, lot# 2065447) was prepared in IMDM (Gibco, cat# 12440-053) with 10% FBS (Gibco, cat# 10082-147). A 2000 ng/ml human-gp96-mFc standard solution was made and 2-fold serial dilutions were performed down to 1 .95 ng/ml. Sample supernatants were loaded onto the ELISA plates starting at a 1 :2 dilution and then 2-fold serial dilutions were performed to a highest dilution of 1 :16. Plates were sealed and incubated for 1 hour at room temperature and then washed 4 times with 1X TBST. The detection antibody, goat anti-mouse IgG (Fc)-FIRP (Jackson Immunoresearch, cat# 115-036-008) was diluted 1 :5,000 in 1X TBST and added to the ELISA plates. Plates were then sealed and incubated in the dark for 1 hour at room temperature. After washing the plates 4 times with 1X TBST, TMB substrate (SeraCare, cat# 5120- 0076) was added to each well and incubated in dark for 15 minutes at room temperature. Reactions were then stopped with 1 N sulfuric acid and plates read on the Biotek ELx800 plate reader. Concentrations of gp96 expressed from each sample were then determined based off the standard curve.
[00248] The amount of mouse OX40L protein expressed by the mHS-130 cells was also determined by ELISA. For each sample to be tested, one million B16F10-Ova9 parental and mHS-130 cells were plated in a 6-well tissue culture plate in a total volume of 1 ml each. Cells were incubated at 37°C with 5% CO2 for 24 hours at which point the supernatants were harvested. Supernatants were then centrifuged at 2500 rpm for 5 minutes to pellet any cell debris. Clarified supernatants were then transferred to new 1 ,5ml tubes and stored at -80°C. Samples collected were from freshly thawed vials of cells.
[00249] To perform the ELISA, 96-well plates were coated with 2.5 ug/ml His-tagged mouse 0X40 protein (Aero Biosystems, cat# OXO-M5228) in PBS. Plates were sealed and stored at 4°C overnight. Plates were then washed 4 times with 1X TBST and then blocked with 1 % BSA (Sigma-Aldrich, cat# A2153) for 1 hour at room temperature. The plates were then washed 4 times with 1X TBST and a mouse lgG1-mouse OX40L standard (Thermo Fisher Scientific, lot# 2214217) was prepared in IMDM with 10% FBS. A 2000 ng/ml mlgG1-mOX40L standard solution was made and 2-fold serial dilutions were performed down to 1 ,95ng/ml. Sample supernatants were loaded onto the ELISA plates and 2-fold serial dilutions were performed. Plates were sealed and incubated for 90 minutes at 37°C and then washed 4 times with 1X TBST. The detection antibody, goat anti-mouse IgG (Fc)-FIRP (Jackson Immunoresearch,
cat# 115-036-008), was diluted 1 :5000 in 1X PBS/0.05% Tween 20/0.1 % BSA and added to the ELISA plates. Plates were then sealed and incubated in the dark for 1 hour at room temperature. After washing the plates 4 times with 1X TBST, TMB substrate was added to each well and incubated in dark for 10 minutes at room temperature. Reactions were then stopped with 1 N sulfuric acid and plates read on the Biotek ELx800 plate reader. Concentrations of OX40L expressed from each sample were then determined based off the standard curve. This protocol is based on human OX40L protocol.
[00250] Table 3: Expression of mouse gp96-lg and OX40L-lg from mHS-110 and mHS-130, respectively, per million cells in a 24-hour period
[00251] Mouse gp96-lg (nanograms/mL per million cells in 24 hrs)
[00252] Mouse OX40L-lg (nanograms/mL per million cells in 24 hrs)
[00253] Table 4: Expression level of active biologic protein per cell type and resulting ratio for each group
[00254] Mice were tail bled consecutively on days 3, 5, 7, 10, 12, and 14 post-immunization and days 17, 19, 21 , 24, 26, 28, 33, 38, 41 , 45, 48, and 54 into heparinized PBS (10 units/ml) and lysed using ACK lysis buffer (150 mM NhUCI, 100 mM KHCO3 and 10 mM EDTA 0.2 Na, pH 7.2) for 3 minutes and neutralized with 1 X PBS. Samples from the OT-I transfer experiments were then centrifuged at 300x g for 5 minutes, supernatant was removed and the cell pellet stained with anti-CD3 (20 pg/ml), anti-CD44 (20 Mg/ml), anti-CD127 (20 pg/ml), anti-KLRG1 (20 pg/ml) and anti-CD8 (5 pg/ml) antibody cocktail made in FACS buffer using Alexa Fluor 700 anti-mouse CD3 (BioLegend cat # 100216), PE-Cy7 anti mouse CD44 antibody (Biolegend Cat # 103030), PE anti-mouse CD127 (BioLegend Cat # 135010), MBL TRP2 tetramer (MBL cat#T03014B, H-2Kb TRP2“SVYDFFVWL” (SEQ ID NO:40)) {used for spleen only), APC anti-mouse KLRG1 (BioLegend Cat # 138412) and Brilliant Violet 421 anti-mouse CD8alpha antibody (BioLegend Cat # 100738) for 30 minutes at 4°C. Consecutive bleeds were collected from the peripheral blood and analysis was performed by flow cytometry on both exogenous, adoptively transferred, OT-1 and endogenous CD8+ and CD4+ T-cells for activation, and short (SLECs) and long term (MPECs) memory markers as outlined in the methods section. Tumor growth kinetics, response rates and infiltrating lymphocytes, were also quantitated.
[00255] Intracellular Cytokine Staining and Flow Cytometry Splenocytes (1 *106) were incubated with synthetic peptides; SIINFEKL, gp100, TRP-1 , TRP-1 -Variant, or TRP-2 in the wells of a 96-well plate at 37°C and 5% C02. Synthetic peptides were added to a final concentration of 0.5 mM with Golgi stop and incubated for 4-10 hours depending on the peptide. Plates were spun, medium was removed, and cells were resuspended with surface markers CD8 and CD3 and incubated at 4°C for 20 minutes. Cells were washed, resuspended in 50 pi of BD Cytofix/Cytoperm, and incubated at 4°C for 20 min before another two washes and staining with anti-IFN-g. Cells were washed once before acquisition and analysis of fluorescence. Analysis was done using FlowJo software (Tree Star Inc.); events were gated for live lymphocytes on FSC x SSC followed by CD8+ cells using CD8 x CD3 and displayed as CD8 x IFN-y.
[00256] B16F10-OVA tumor challenge and volume calculations : Melanoma B16F10 cells were harvested and resuspended at a concentration of 5x105 cells/100 mI in a volume of 80 mI HBSS and 20 mI Matrigel. C57BL/6 mice were subcutaneously injected with 100 mI of B16F10 cells (5x105 cells/mouse) on the inner abdomen 29 days post-OT-1 transfer and 28 days post-primary vaccination, designated "day 28”, as shown in the study design (FIG. 13). The tumor size was measured and documented every 3 days with a caliper, starting on day 7, and calculated using the formula (AxB; A as the largest and B as the smallest diameter of tumor). Tumor growth was documented as standard error mean. To record the survival of the tumor-bearing mice, either natural death or a tumor volume greater than 450 mm2 leading to death was counted as death. Each experimental group included five animals.
[00257] Tumor tissue digestion for Tumor Infiltrating Lymphocytes (TILs): The MACS Miltenyl Biotec tumor dissociation kit was used for this procedure (cat# 130096-730).
[00258] ELISPOT Assay: Splenocytes were harvested and red blood cell lysis buffer (cat#36858500, Roche) was used to eliminate red blood cells. Cells were washed in IMDM medium and pelleted. Counted cells were resuspended in IMDM with 10% FBS. Each ELISPOT well received 1 million cells, in a total volume of 200 mI. Treatments included the use of B16F10-OVA parental line lysates (vial of 10 million cells; freeze-thawed three times) at a 10-fold dilution, immunodominant epitopes for B16F10 tumors: "gp100/pmel” (EGSRNQDWL (SEQ ID NO:38)), "TRP-1/gp75” (TWHRYHLL (SEQ ID NO:39)), TRP-2 (SVYDFFVWL (SEQ ID NO:40)) and the MHC-I restricted peptide for H2b haplotype for OVA (SIINFEKL (SEQ ID NO:41)). All peptides were used at a final concentration of 10 ug/mL. PMA/lonomycin was used as a positive control at "1 to 400 dilution” 100 nM of PMA, 1.6 mM ionomycin (500x stock from eBioscience, cat#00-4970-03, and whole chicken ovalbumin protein "OVA” at 250 pg/ml.
[00259] Interferon-gamma capture ELISPOT assay was carried out as previously published (Klinman et al., Current Protocols in Immunology. 1994 June (1 ): 6.19.1-6.19.8). Briefly, MAIP N45 Milipore 96- well filtration plates were coated with anti-mouse IFN-g purified monoclonal antibody (clone AN18) in PBS
saline at 10 mo/hhί, 50 mI/well, at 4°C, overnight. The next day, cells were washed in IMEM medium plus 10% FBS at 200 ul/well, washed three times, then blocked for 1 hour at 25°C. Cells were processed as described above and 1 million or 100,000 splenocytes were added to each well and treated per the description above with various peptide stimulations. Each plate was incubated in a 37°C, 5% CO2, non vibrating incubator for 16 hrs or overnight. At the end of the incubation, cells were flicked into the sink to stop the culture and washed with PBS + 0.1 % Tween-20 three times and blotted hard onto paper towels to get rid of residual liquid after the last wash. Secondary biotinylated antibody was added at 50 mI per well, 4 pg/mL of anti-IFN-g (clone AN18), incubated at 25°C for 2 hrs. Plates were then washed again with PBS+0.1 % Tween-20 three times. Peroxidase-conjugated streptavidin from Jackson research labs was then added at a 1 to 1000 dilution from stock concentration in PBS + 0.1 % Tween-20 + 2% BSA, 50 mI per well, 30-minute incubation at 4°C. Plates were then washed with PBS + 0.1 % Tween three times. The plastic backing of the ELISPOT plate was removed and the entire plate was submerged and soaked in PBS + 0.1 % Tween-20 for 1 hour at 25°C. The plate was then washed in PBS to remove the tween, then 100 mI of the AEC substrate (Vector kit) added in 100 mM of TRIS buffer at pH 8.2 was added to each well to allow for development of spots. At the end of the incubation, 10-20 minutes, all plates incubated for the same time, the reaction as stopped by rinsing the plates with tap water and air dried.
[00260] Plates were scanned and counted in an AID Autoimmun Diagnostika GMBPI iSpot monochromatic ELISPOT reader (2018). Counting parameters for each plate remained consistent across all plates that were developed together as to prevent bias, as spot development, density and magnitude is influence by the operator, reagent lots and final substrate development time.
[00261] Experimental Results
[00262] As shown in FIGS. 14, 15A, 15B, 15C, and 15D, priming with mHS-1 10 (gp96-lg) and mPIS- 130 (OX40L-lg) produced a primary immune response and cellular expansion of OT-1 cells (anti-OVA, CD8+ TCR transgenic T-cells) in a dose and ratio dependent manner.
[00263] FIG. 14 illustrates anti-tumor CD8+ OT-I T cell expansion in the peripheral blood with prime and boost Immunization of different ratios and dose combinations of mHS-110 and mPIS-130. Recipient mice were injected with mHS-110 and mHS-130 at different ratios and doses of gp96-lg to OX40L-lg. OT-I GFP+ CD8+ T cells were analyzed in the blood on days 0-54 days post-vaccination. Mice were boosted on day 14 with the same ratios of mHS-1 10 and mHS-130 as in the primary phase, and OT-I GFP+ CD8+ T cells were analyzed in the blood days post-challenge. As shown, the ratio 1 :1.3 ratio of mHS-1 10 to mHS-130 (high dose) results in the peak of CD8+ OT-l T cell expansion at day 7 and remains higher than other dose ratios through day 54.
[00264] FIG. 15A illustrates gating strategy for flow cytometry experiments of the study of FIG. 13.
Recipient mice were injected with a mHS-110 and mHS-130 at different ratios and doses of gp96-lg to OX40L-lg. OT-I GFP+ CD8+ T cells were analyzed in the blood on days 0-54 days post-vaccination. Mice were boosted on day 14 and 31 with the same ratios of mHS-110 and mHS-130 as in the primary phase, and OT-I GFP+ CD8+ T cells were analyzed in the blood days post-challenge. Tumor was provided on day 28.
[00265] FIG. 15B, illustrating expansion of OT-1 cells on days 7 and 17, shows that the dose ratio 1 to 1.3 provided the best response with the greatest expansion seen for the high dose, 339 ng gp96 to 441 ng OX40L. In particular, upon the primary immunization, there was a dramatic, nearly a three-fold increase in OT-1 cells after adding OX40L to gp96 (38 ng gp96 vs. 50 ng group; for the 1 to 1.3 ratio) when compared to mHS-1 10 (38 ng gp96) alone, as shown in FIG. 15B. There was a trend decrease in expansion of OT-1 cells that indirectly correlated with an increase in OX40L-lg to gp96-lg, as shown for day 7 in FIG. 15B. This trend was observed for other days (from day 7 to day 26) but is most pronounced at the primary immunization peak of day 7. Boosting on day 14 produced an increase in cellular expansion, and, among the studied dose ratio groups, the observed expansion was not pronounced for the 1 to 1.3 ratio (FIG. 15B, day 17). The groups that harbored the most OT-1 cells just prior to the boosting on day 14, showed the best and most rapid response shown on day 17 through day 26, with the same dose trend between the ratios and groups, especially as it correlates to OX40L-lg, as shown in FIG. 15C. The 1 to 1.3 ratio of gp96-lg to OX40L-lg maintained the best expansion of CD8+ OT-I T-cells through the end of the study, as shown in FIG. 15D illustrating the results for days 45, 48, and 54.
[00266] In the present study, activation and key memory markers were also measured for the studied peripheral blood populations. Flow cytometry gating strategy for memory markers KLRG1 and IL-7R is shown in FIG. 16. Further, as shown in FIG. 17, significant changes in CD8+ memory precursor effector cells (MPECs) (KLRG110 IL-7Rhi) and short-lived effector cells (SLECs) (KLRGhi IL-7R10) were observed for endogenous cell populations on day 7. Short-lived effector cells increased directly proportional to gp96-lg exposure; but indirectly proportional to OX40L-lg (FIG. 17). This suggests that increasing OX40L may produce better CD8+ T-cell expansion. These data support an approach of combining both gp96 and OX40L in a single vaccine.
[00267] In addition, similar to what was observed with OT-1 T-cell expansion in FIGS. 15B-15D, greater OX40L to less gp96 stimulation results in a more robust cellular response, resulting in greater SLEC formation (FIG. 17, day 7, endogenous CD8+ T-cells). Formation of SLECs correlated with increased activation, as shown by the upregulation of adhesion molecule, CD44, on endogenous CD8+ T-cell populations (FIG. 18, day 7). Expression of CD44, which appears on antigen stimulated T-cells to allow entry to target tissues, is highly elevated on endogenous populations and trends with the increased dose
levels of OX40L relative to gp96.
[00268] On day 28, B16F10-OVA tumors were given subcutaneously, and tumor delay challenge began. Tumor growth inhibition for each group tested is shown in FIG. 19. Significant differences, as compared to mHS-1 10 (38 ng gp96-lg) alone, measured by 1-way ANOVA statistical test were observed for the 339 ng to 441 ng group (1 to 1.3, high, gp96 to OX40L), 339 ng to 848 ng (1 to 2.5, high, gp96 to OX40L), and 339 ng to 1695 ng (1 to 5, high, gp96 to OX40L) (FIG. 19, ****p<0.0001 ). End of study (day 55) tumor weights, grouped (FIG. 20, left) and individual (FIG. 20, right) agree with the caliper measurements (FIG. 19).
[00269] Tumor-specific, TRP2 tetramer positive CD8+ T-cells increased with treatment in a dose- dependent manner and showed a significant increase over treatment with 38 ng of gp96-lg via mHS-110 alone (FIG. 21) As for the transferred CD8+ OT-1 + eGFP cells, at day 55, only the 1 to 1 .3 ratio of 339 ng gp96-lg to 441 ng OX40L-lg dose (high dose) showed any presence of expanded subsets on day 55 in both the spleen and blood, as shown in FIG. 22.
[00270] FIG. 23 illustrates an increased percentage of exhausted CD4+ T-cells staining positive for PD-1 in the spleen (% of CD3+ CD4+ PD-1 + T cells) on day 55. The percentage of effector memory CD4+ T-cells in the spleen of treated and tumor-burden mice changed with vaccination, in an OX40L dose-dependent manner, as shown in FIG. 24. This increase in the percentage of activated CD4+ T- cells correlated with an increase proportion of CD4+ T-cell infiltrating the tumor (TILs), and was dose dependent (see FIG. 26).
[00271] The present study demonstrates that 1) both CD4+ and CD8+ T-cell subsets are expanded with gp96/OX40L-lg co-vaccinations that correlate with increased TIL percentages and tumor growth inhibition; 2) the best dose combination for long-term survival and expansion of tumor specific CD8+ T- cells cells is the 1 to 1.3 ratio at a dose of 339 ng of gp96-lg to 441 ng of OX40L-lg; and 3) a dose determines tumor growth inhibition, with 38 ng of gp96-lg to 50 ng of OX40L-lg being the no-observed effect level (NOEL) and 1 13 ng gp96-lg to 147 ng of OX40L-lg being the minimum active biological effect level (MABEL) for this dose combination, in mice.
OTHER EMBODIMENTS
[00272] It is to be understood that while the disclosure has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the disclosure, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.
INCORPORATION BY REFERENCE
[00273] All patents and publications referenced herein are hereby incorporated by reference in their
entireties. The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. As used herein, all headings are simply for organization and are not intended to limit the disclosure in anyway.
Claims
1. A method for treating a patient comprising administering to the patient an effective amount of a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein,
wherein the patient is undergoing a treatment with a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject.
2. A method for treating a patient comprising administering to the patient an effective amount of a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein, wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject, and
wherein the patient is undergoing a treatment with a first cell comprising an expression vector comprising, a nucleotide sequence that encodes a secretable vaccine protein.
3. A method for treating a patient comprising administering to the patient an effective amount of
(a) a first cell comprising an expression vector comprising a nucleotide sequence that encodes a secretable vaccine protein and
(b) a second cell comprising an expression vector comprising a nucleotide sequence that encodes a T cell costimulatory fusion protein and wherein the T cell costimulatory fusion protein enhances activation of antigen-specific T cells when administered to the subject.
4. The method of any one of claims 1 -3, wherein the secretable vaccine protein is a secretable gp96-lg fusion protein which optionally lacks the gp96 KDEL (SEQ ID NO:3) sequence.
5. The method of claim 4, wherein the Ig tag in the gp96-lg fusion protein comprises the Fc region of human lgG1 , lgG2, lgG3, lgG4, IgM, IgA, or IgE.
6. The method of any one of claims 1-3 wherein the T cell costimulatory fusion protein is OX40L-lg, or a portion thereof that binds to 0X40.
7. The method of any one of claims 1-3, wherein the T cell costimulatory fusion protein is ICOSL-lg, or a portion thereof that binds to ICOS.
8. The method of any one of claims 1-3, wherein the T cell costimulatory fusion protein is 4-1 BBL-lg, or a portion thereof that binds to 4-1 BBR.
9. The method of any one of claims 1-3, wherein the T cell costimulatory fusion protein is TL1 A-lg, or a portion thereof that binds to TNFRSF25.
10. The method of any one of claims 1-3, wherein the T cell costimulatory fusion protein is GITRL-lg, or a portion thereof that binds to GITR.
11.The method of any one of claims 1-3, wherein the T cell costimulatory fusion protein is CD40L-lg, or a portion thereof that binds to CD40.
12. The method of any one of claims 1-3, wherein the T cell costimulatory fusion protein is CD70-lg, or a portion thereof that binds to CD27.
13. The method of any one of claims 6-12, wherein the Ig tag in the T cell costimulatory fusion protein comprises the Fc region of human lgG1 , lgG2, lgG3, lgG4, IgM, IgA, or IgE.
14. The method of any one of claims 1-13, wherein the expression vector is incorporated into a virus or virus-like particle.
15. The method of any one of claims 1-13, wherein the expression vector is incorporated into a human tumor cell.
16. The method of any one of claims 1-13, wherein the patient is a human cancer patient.
17. The method of claim 15, wherein administration to the human patient increases the activation or proliferation of tumor antigen specific T cells in the patient.
18. The method of any one of claims 1 , 2, 3 or 17, wherein the activation or proliferation of tumor antigen specific T cells in the patient is increased by at least 25 percent as compared to the level of activation or proliferation of tumor antigen specific T cells in the patient prior to the administration.
19. The method of claim 18, comprising administering in combination with an agent that inhibits immunosuppressive molecules produced by tumor cells.
20. The method of claim 19, wherein the agent is an antibody against PD-1.
21. The method of claim 20, wherein the antibody against PD-1 is selected from nivolumab, pembrolizumab, pidilizumab, cemiplimab, AGEN2034, AMP-224, AMP-514, PDR001.
22. The method of any one of claims 1-21 , wherein the patient is a human with an acute or chronic infection.
23. The method of claim 22 wherein the acute or chronic infection is an infection by hepatitis C virus, hepatitis B virus, human immunodeficiency virus, or malaria.
24. The method of claim 22, wherein administration to the human patient stimulates the activation or proliferation of pathogenic antigen specific T cells.
25. The method of any one of claims 1-3, wherein the T cell costimulatory molecule enhances the activation of antigen-specific T cells in the subject to a greater level than gp96-lg vaccination alone.
26. The method of any one of the preceding claims, wherein the ratio of the secretable vaccine protein to the T cell costimulatory fusion protein is about 1 :1.
27. The method of any one of claims 1-25, wherein the ratio of the secretable vaccine protein to the T cell costimulatory fusion protein is about 1 :1.3.
28. The method of any one of claims 1-25, wherein the ratio of the secretable vaccine protein to the T cell costimulatory fusion protein is about 1 :10.
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US17/277,871 US20210346486A1 (en) | 2018-10-01 | 2019-10-01 | Combination cell-based therapies |
EP19869950.6A EP3860647A4 (en) | 2018-10-01 | 2019-10-01 | Combination cell-based therapies |
CA3113454A CA3113454A1 (en) | 2018-10-01 | 2019-10-01 | Combination cell-based therapies |
CN201980079182.7A CN113164574A (en) | 2018-10-01 | 2019-10-01 | Cell-based combination therapy |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201862739814P | 2018-10-01 | 2018-10-01 | |
US62/739,814 | 2018-10-01 | ||
US201962807783P | 2019-02-20 | 2019-02-20 | |
US62/807,783 | 2019-02-20 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2020072395A1 true WO2020072395A1 (en) | 2020-04-09 |
Family
ID=70055046
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2019/053925 WO2020072395A1 (en) | 2018-10-01 | 2019-10-01 | Combination cell-based therapies |
Country Status (5)
Country | Link |
---|---|
US (1) | US20210346486A1 (en) |
EP (1) | EP3860647A4 (en) |
CN (1) | CN113164574A (en) |
CA (1) | CA3113454A1 (en) |
WO (1) | WO2020072395A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2022155212A1 (en) * | 2021-01-13 | 2022-07-21 | Heat Biologics, Inc. | Cell-fusion based immune agents |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
MX2017010159A (en) * | 2015-02-06 | 2017-12-18 | Heat Biologics Inc | Vector co-expressing vaccine and costimulatory molecules. |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016127015A1 (en) * | 2015-02-06 | 2016-08-11 | Heat Biologics, Inc. | Vector co-expressing vaccine and costimulatory molecules |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10196435B2 (en) * | 2013-11-18 | 2019-02-05 | University Of Southern California | OX40L fusion protein for the immunotherapy of tumors of veterinary animals |
WO2018187260A1 (en) * | 2017-04-04 | 2018-10-11 | Heat Biologics, Inc. | Intratumoral vaccination |
-
2019
- 2019-10-01 CN CN201980079182.7A patent/CN113164574A/en active Pending
- 2019-10-01 CA CA3113454A patent/CA3113454A1/en active Pending
- 2019-10-01 EP EP19869950.6A patent/EP3860647A4/en not_active Withdrawn
- 2019-10-01 WO PCT/US2019/053925 patent/WO2020072395A1/en unknown
- 2019-10-01 US US17/277,871 patent/US20210346486A1/en not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016127015A1 (en) * | 2015-02-06 | 2016-08-11 | Heat Biologics, Inc. | Vector co-expressing vaccine and costimulatory molecules |
US20160250322A1 (en) * | 2015-02-06 | 2016-09-01 | Heat Biologics, Inc. | Vector co-expressing vaccine and costimulatory molecules |
Non-Patent Citations (2)
Title |
---|
FROMM, G ET AL.: "Gp96-Ig/Costimulator (OX40L, ICOSL, or 4-1 BBL) Combination Vaccine Improves T- cell Priming and Enhances Immunity, Memory, and Tumor Elimination", CANCER IMMUNOLOGY RESEARCH, vol. 4, no. 9, 2 September 2016 (2016-09-02), pages 766 - 778, XP055699340, [retrieved on 20160630] * |
See also references of EP3860647A4 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2022155212A1 (en) * | 2021-01-13 | 2022-07-21 | Heat Biologics, Inc. | Cell-fusion based immune agents |
Also Published As
Publication number | Publication date |
---|---|
EP3860647A1 (en) | 2021-08-11 |
EP3860647A4 (en) | 2022-10-19 |
US20210346486A1 (en) | 2021-11-11 |
CA3113454A1 (en) | 2020-04-09 |
CN113164574A (en) | 2021-07-23 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20210000945A1 (en) | Vector co-expressing vaccine and costimulatory molecules | |
US10988517B2 (en) | Heterodimeric proteins for modulating gamma delta T cells | |
US20230303659A1 (en) | Intratumoral vaccination | |
US20230020258A1 (en) | Heterodimeric proteins for modulating gamma delta t cells | |
WO2020072395A1 (en) | Combination cell-based therapies | |
US20220298252A1 (en) | Methods of treating cancer using tnfrsf25 antibodies | |
WO2022155212A1 (en) | Cell-fusion based immune agents |
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: 19869950 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 3113454 Country of ref document: CA |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 2019869950 Country of ref document: EP Effective date: 20210503 |