US20050202010A1 - Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor - Google Patents
Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor Download PDFInfo
- Publication number
- US20050202010A1 US20050202010A1 US10/927,494 US92749404A US2005202010A1 US 20050202010 A1 US20050202010 A1 US 20050202010A1 US 92749404 A US92749404 A US 92749404A US 2005202010 A1 US2005202010 A1 US 2005202010A1
- Authority
- US
- United States
- Prior art keywords
- mif
- subject
- treating
- preventing
- need
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 230000000747 cardiac effect Effects 0.000 title claims abstract description 292
- 238000000034 method Methods 0.000 title claims abstract description 196
- 108010048043 Macrophage Migration-Inhibitory Factors Proteins 0.000 title description 59
- 102000009073 Macrophage Migration-Inhibitory Factors Human genes 0.000 title description 59
- 238000011282 treatment Methods 0.000 title description 47
- 108010050608 Myocardial Depressant Factor Proteins 0.000 title description 9
- 238000004166 bioassay Methods 0.000 title description 3
- 230000004064 dysfunction Effects 0.000 claims abstract description 264
- 208000014674 injury Diseases 0.000 claims abstract description 155
- 239000003112 inhibitor Substances 0.000 claims abstract description 151
- 208000027418 Wounds and injury Diseases 0.000 claims abstract description 148
- 230000006378 damage Effects 0.000 claims abstract description 114
- 230000004217 heart function Effects 0.000 claims abstract description 77
- 206010000891 acute myocardial infarction Diseases 0.000 claims abstract description 71
- 239000003937 drug carrier Substances 0.000 claims abstract description 52
- 230000036459 cardiodepression Effects 0.000 claims abstract description 48
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 37
- 230000000694 effects Effects 0.000 claims description 110
- 230000002107 myocardial effect Effects 0.000 claims description 109
- 239000000203 mixture Substances 0.000 claims description 93
- 102100030595 HLA class II histocompatibility antigen gamma chain Human genes 0.000 claims description 54
- 101001082627 Homo sapiens HLA class II histocompatibility antigen gamma chain Proteins 0.000 claims description 54
- 150000003384 small molecules Chemical class 0.000 claims description 42
- 210000000107 myocyte Anatomy 0.000 claims description 40
- 150000003839 salts Chemical class 0.000 claims description 39
- 239000003795 chemical substances by application Substances 0.000 claims description 27
- 239000000651 prodrug Substances 0.000 claims description 15
- 229940002612 prodrug Drugs 0.000 claims description 15
- 241000699670 Mus sp. Species 0.000 description 177
- 239000002158 endotoxin Substances 0.000 description 163
- 229920006008 lipopolysaccharide Polymers 0.000 description 142
- 210000002216 heart Anatomy 0.000 description 103
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 92
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 92
- 102000004127 Cytokines Human genes 0.000 description 83
- 108090000695 Cytokines Proteins 0.000 description 83
- 206010040047 Sepsis Diseases 0.000 description 50
- 210000002966 serum Anatomy 0.000 description 50
- 235000002639 sodium chloride Nutrition 0.000 description 46
- LOGFVTREOLYCPF-KXNHARMFSA-N (2s,3r)-2-[[(2r)-1-[(2s)-2,6-diaminohexanoyl]pyrrolidine-2-carbonyl]amino]-3-hydroxybutanoic acid Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]1CCCN1C(=O)[C@@H](N)CCCCN LOGFVTREOLYCPF-KXNHARMFSA-N 0.000 description 38
- 102000003777 Interleukin-1 beta Human genes 0.000 description 38
- 108090000193 Interleukin-1 beta Proteins 0.000 description 38
- 108060008683 Tumor Necrosis Factor Receptor Proteins 0.000 description 38
- 102000003298 tumor necrosis factor receptor Human genes 0.000 description 38
- 210000004379 membrane Anatomy 0.000 description 36
- 239000012528 membrane Substances 0.000 description 36
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 32
- 230000014509 gene expression Effects 0.000 description 30
- 230000005764 inhibitory process Effects 0.000 description 29
- 210000005240 left ventricle Anatomy 0.000 description 29
- 230000006870 function Effects 0.000 description 28
- 210000001519 tissue Anatomy 0.000 description 28
- 239000000872 buffer Substances 0.000 description 27
- 238000001727 in vivo Methods 0.000 description 26
- 230000011664 signaling Effects 0.000 description 26
- 206010040070 Septic Shock Diseases 0.000 description 25
- 238000002560 therapeutic procedure Methods 0.000 description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 25
- 241001465754 Metazoa Species 0.000 description 24
- 238000002592 echocardiography Methods 0.000 description 24
- 230000036303 septic shock Effects 0.000 description 24
- 150000001875 compounds Chemical class 0.000 description 23
- 108020004999 messenger RNA Proteins 0.000 description 23
- 108090001005 Interleukin-6 Proteins 0.000 description 22
- 102000004889 Interleukin-6 Human genes 0.000 description 22
- 241000699666 Mus <mouse, genus> Species 0.000 description 22
- 210000004413 cardiac myocyte Anatomy 0.000 description 22
- 210000000952 spleen Anatomy 0.000 description 22
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 21
- 239000000523 sample Substances 0.000 description 21
- 238000004904 shortening Methods 0.000 description 21
- 241000282414 Homo sapiens Species 0.000 description 20
- 230000007423 decrease Effects 0.000 description 20
- 239000000499 gel Substances 0.000 description 20
- 230000007246 mechanism Effects 0.000 description 20
- 238000002360 preparation method Methods 0.000 description 20
- 108090000623 proteins and genes Proteins 0.000 description 20
- 201000010099 disease Diseases 0.000 description 19
- 210000004185 liver Anatomy 0.000 description 19
- 238000006386 neutralization reaction Methods 0.000 description 19
- 230000004224 protection Effects 0.000 description 19
- 230000003111 delayed effect Effects 0.000 description 17
- 230000002401 inhibitory effect Effects 0.000 description 17
- 230000010412 perfusion Effects 0.000 description 17
- 238000003556 assay Methods 0.000 description 16
- 238000002474 experimental method Methods 0.000 description 16
- 102000004169 proteins and genes Human genes 0.000 description 16
- 238000001262 western blot Methods 0.000 description 16
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 15
- 239000003085 diluting agent Substances 0.000 description 15
- 230000002265 prevention Effects 0.000 description 15
- 230000004044 response Effects 0.000 description 15
- 239000000243 solution Substances 0.000 description 15
- 102000007469 Actins Human genes 0.000 description 14
- 108010085238 Actins Proteins 0.000 description 14
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 14
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 14
- 102100037850 Interferon gamma Human genes 0.000 description 14
- 108010074328 Interferon-gamma Proteins 0.000 description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 14
- 230000028327 secretion Effects 0.000 description 14
- 239000003826 tablet Substances 0.000 description 14
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 13
- 238000004458 analytical method Methods 0.000 description 13
- 230000000994 depressogenic effect Effects 0.000 description 13
- 238000003364 immunohistochemistry Methods 0.000 description 13
- 239000007788 liquid Substances 0.000 description 13
- 108010057531 macrophage migration inhibitory factor receptor Proteins 0.000 description 13
- 125000001424 substituent group Chemical group 0.000 description 13
- 102000003810 Interleukin-18 Human genes 0.000 description 12
- 108090000171 Interleukin-18 Proteins 0.000 description 12
- SXEHKFHPFVVDIR-UHFFFAOYSA-N [4-(4-hydrazinylphenyl)phenyl]hydrazine Chemical compound C1=CC(NN)=CC=C1C1=CC=C(NN)C=C1 SXEHKFHPFVVDIR-UHFFFAOYSA-N 0.000 description 12
- 230000001154 acute effect Effects 0.000 description 12
- 238000000540 analysis of variance Methods 0.000 description 12
- 210000005003 heart tissue Anatomy 0.000 description 12
- 206010001052 Acute respiratory distress syndrome Diseases 0.000 description 11
- 108010065805 Interleukin-12 Proteins 0.000 description 11
- 102000013462 Interleukin-12 Human genes 0.000 description 11
- 239000004480 active ingredient Substances 0.000 description 11
- 210000004027 cell Anatomy 0.000 description 11
- 230000003247 decreasing effect Effects 0.000 description 11
- 235000019441 ethanol Nutrition 0.000 description 11
- 230000002757 inflammatory effect Effects 0.000 description 11
- 238000004519 manufacturing process Methods 0.000 description 11
- 102000005962 receptors Human genes 0.000 description 11
- 108020003175 receptors Proteins 0.000 description 11
- 238000011740 C57BL/6 mouse Methods 0.000 description 10
- 238000000636 Northern blotting Methods 0.000 description 10
- 208000013616 Respiratory Distress Syndrome Diseases 0.000 description 10
- 201000000028 adult respiratory distress syndrome Diseases 0.000 description 10
- 210000000038 chest Anatomy 0.000 description 10
- 239000010408 film Substances 0.000 description 10
- 238000000338 in vitro Methods 0.000 description 10
- 230000003472 neutralizing effect Effects 0.000 description 10
- 238000001543 one-way ANOVA Methods 0.000 description 10
- 230000009885 systemic effect Effects 0.000 description 10
- 241000283707 Capra Species 0.000 description 9
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- 241000283973 Oryctolagus cuniculus Species 0.000 description 9
- 238000010162 Tukey test Methods 0.000 description 9
- 239000002253 acid Substances 0.000 description 9
- 238000006243 chemical reaction Methods 0.000 description 9
- 239000003153 chemical reaction reagent Substances 0.000 description 9
- 230000003205 diastolic effect Effects 0.000 description 9
- 238000002347 injection Methods 0.000 description 9
- 239000007924 injection Substances 0.000 description 9
- -1 isoxazoline compounds Chemical class 0.000 description 9
- 210000002540 macrophage Anatomy 0.000 description 9
- 238000005259 measurement Methods 0.000 description 9
- 239000000546 pharmaceutical excipient Substances 0.000 description 9
- 239000000843 powder Substances 0.000 description 9
- 210000003491 skin Anatomy 0.000 description 9
- 230000004083 survival effect Effects 0.000 description 9
- 230000001225 therapeutic effect Effects 0.000 description 9
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 8
- 241000700159 Rattus Species 0.000 description 8
- 229920002472 Starch Polymers 0.000 description 8
- 238000003119 immunoblot Methods 0.000 description 8
- 230000006872 improvement Effects 0.000 description 8
- 210000003734 kidney Anatomy 0.000 description 8
- 239000000314 lubricant Substances 0.000 description 8
- 208000010125 myocardial infarction Diseases 0.000 description 8
- 230000000770 proinflammatory effect Effects 0.000 description 8
- 239000011780 sodium chloride Substances 0.000 description 8
- 235000019698 starch Nutrition 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 238000002965 ELISA Methods 0.000 description 7
- 206010053879 Sepsis syndrome Diseases 0.000 description 7
- 206010051379 Systemic Inflammatory Response Syndrome Diseases 0.000 description 7
- 108010060804 Toll-Like Receptor 4 Proteins 0.000 description 7
- 102000008233 Toll-Like Receptor 4 Human genes 0.000 description 7
- 239000002585 base Substances 0.000 description 7
- 239000011230 binding agent Substances 0.000 description 7
- 239000002775 capsule Substances 0.000 description 7
- 230000001684 chronic effect Effects 0.000 description 7
- 230000001404 mediated effect Effects 0.000 description 7
- 230000005012 migration Effects 0.000 description 7
- 238000013508 migration Methods 0.000 description 7
- 229910052757 nitrogen Inorganic materials 0.000 description 7
- 230000008506 pathogenesis Effects 0.000 description 7
- 238000011084 recovery Methods 0.000 description 7
- 239000007787 solid Substances 0.000 description 7
- 230000006641 stabilisation Effects 0.000 description 7
- 238000011105 stabilization Methods 0.000 description 7
- AYEKOFBPNLCAJY-UHFFFAOYSA-O thiamine pyrophosphate Chemical compound CC1=C(CCOP(O)(=O)OP(O)(O)=O)SC=[N+]1CC1=CN=C(C)N=C1N AYEKOFBPNLCAJY-UHFFFAOYSA-O 0.000 description 7
- 230000008733 trauma Effects 0.000 description 7
- 230000002861 ventricular Effects 0.000 description 7
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 6
- 0 C*(*)(C(*)(*)C*)N Chemical compound C*(*)(C(*)(*)C*)N 0.000 description 6
- 238000011746 C57BL/6J (JAX™ mouse strain) Methods 0.000 description 6
- 244000025254 Cannabis sativa Species 0.000 description 6
- 108020004414 DNA Proteins 0.000 description 6
- 208000037487 Endotoxemia Diseases 0.000 description 6
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 6
- 241000282412 Homo Species 0.000 description 6
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 241001529936 Murinae Species 0.000 description 6
- 108010057466 NF-kappa B Proteins 0.000 description 6
- 102000003945 NF-kappa B Human genes 0.000 description 6
- 206010028980 Neoplasm Diseases 0.000 description 6
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 6
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 6
- 239000011543 agarose gel Substances 0.000 description 6
- 230000008901 benefit Effects 0.000 description 6
- 229960002685 biotin Drugs 0.000 description 6
- 239000011616 biotin Substances 0.000 description 6
- 230000003684 cardiac depression Effects 0.000 description 6
- 230000005961 cardioprotection Effects 0.000 description 6
- 210000004351 coronary vessel Anatomy 0.000 description 6
- 239000008367 deionised water Substances 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 239000003862 glucocorticoid Substances 0.000 description 6
- 208000019622 heart disease Diseases 0.000 description 6
- 230000001976 improved effect Effects 0.000 description 6
- 239000007928 intraperitoneal injection Substances 0.000 description 6
- 238000011813 knockout mouse model Methods 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 235000013336 milk Nutrition 0.000 description 6
- 239000008267 milk Substances 0.000 description 6
- 210000004080 milk Anatomy 0.000 description 6
- 230000007935 neutral effect Effects 0.000 description 6
- 230000036542 oxidative stress Effects 0.000 description 6
- 239000012188 paraffin wax Substances 0.000 description 6
- 230000037361 pathway Effects 0.000 description 6
- 230000036470 plasma concentration Effects 0.000 description 6
- 229940044551 receptor antagonist Drugs 0.000 description 6
- 239000002464 receptor antagonist Substances 0.000 description 6
- 230000001105 regulatory effect Effects 0.000 description 6
- 230000035939 shock Effects 0.000 description 6
- 230000019491 signal transduction Effects 0.000 description 6
- 238000007619 statistical method Methods 0.000 description 6
- 230000002123 temporal effect Effects 0.000 description 6
- 238000012546 transfer Methods 0.000 description 6
- 239000003656 tris buffered saline Substances 0.000 description 6
- 239000008096 xylene Substances 0.000 description 6
- 206010069140 Myocardial depression Diseases 0.000 description 5
- 102000004316 Oxidoreductases Human genes 0.000 description 5
- 108090000854 Oxidoreductases Proteins 0.000 description 5
- 230000002238 attenuated effect Effects 0.000 description 5
- 239000011324 bead Substances 0.000 description 5
- 230000033228 biological regulation Effects 0.000 description 5
- 230000000903 blocking effect Effects 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 230000009091 contractile dysfunction Effects 0.000 description 5
- 229910021641 deionized water Inorganic materials 0.000 description 5
- 238000010790 dilution Methods 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- 239000007884 disintegrant Substances 0.000 description 5
- 239000002552 dosage form Substances 0.000 description 5
- 239000000975 dye Substances 0.000 description 5
- 239000000839 emulsion Substances 0.000 description 5
- 230000002255 enzymatic effect Effects 0.000 description 5
- 239000012634 fragment Substances 0.000 description 5
- 229910052739 hydrogen Inorganic materials 0.000 description 5
- 210000002865 immune cell Anatomy 0.000 description 5
- 239000007927 intramuscular injection Substances 0.000 description 5
- 238000010255 intramuscular injection Methods 0.000 description 5
- 238000007912 intraperitoneal administration Methods 0.000 description 5
- 210000004072 lung Anatomy 0.000 description 5
- 210000003205 muscle Anatomy 0.000 description 5
- 210000004165 myocardium Anatomy 0.000 description 5
- 231100000252 nontoxic Toxicity 0.000 description 5
- 230000003000 nontoxic effect Effects 0.000 description 5
- 210000000056 organ Anatomy 0.000 description 5
- 229910052760 oxygen Inorganic materials 0.000 description 5
- 239000013612 plasmid Substances 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 239000008107 starch Substances 0.000 description 5
- 238000010254 subcutaneous injection Methods 0.000 description 5
- 239000007929 subcutaneous injection Substances 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 238000013518 transcription Methods 0.000 description 5
- 230000035897 transcription Effects 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 4
- 108010006533 ATP-Binding Cassette Transporters Proteins 0.000 description 4
- 102000005416 ATP-Binding Cassette Transporters Human genes 0.000 description 4
- 206010002091 Anaesthesia Diseases 0.000 description 4
- 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 4
- 108010010803 Gelatin Proteins 0.000 description 4
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 4
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 4
- 206010061216 Infarction Diseases 0.000 description 4
- 108010002350 Interleukin-2 Proteins 0.000 description 4
- 102000000588 Interleukin-2 Human genes 0.000 description 4
- 108090000978 Interleukin-4 Proteins 0.000 description 4
- 102000004388 Interleukin-4 Human genes 0.000 description 4
- 108010002616 Interleukin-5 Proteins 0.000 description 4
- 102000000743 Interleukin-5 Human genes 0.000 description 4
- 239000012839 Krebs-Henseleit buffer Substances 0.000 description 4
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 4
- 239000002033 PVDF binder Substances 0.000 description 4
- 229920001213 Polysorbate 20 Polymers 0.000 description 4
- 102000013394 Troponin I Human genes 0.000 description 4
- 108010065729 Troponin I Proteins 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 230000037005 anaesthesia Effects 0.000 description 4
- 206010003119 arrhythmia Diseases 0.000 description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 4
- RMRJXGBAOAMLHD-IHFGGWKQSA-N buprenorphine Chemical compound C([C@]12[C@H]3OC=4C(O)=CC=C(C2=4)C[C@@H]2[C@]11CC[C@]3([C@H](C1)[C@](C)(O)C(C)(C)C)OC)CN2CC1CC1 RMRJXGBAOAMLHD-IHFGGWKQSA-N 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 210000000172 cytosol Anatomy 0.000 description 4
- 230000006735 deficit Effects 0.000 description 4
- 238000000326 densiometry Methods 0.000 description 4
- 230000001419 dependent effect Effects 0.000 description 4
- 238000001962 electrophoresis Methods 0.000 description 4
- 235000013861 fat-free Nutrition 0.000 description 4
- 210000001035 gastrointestinal tract Anatomy 0.000 description 4
- 229920000159 gelatin Polymers 0.000 description 4
- 235000019322 gelatine Nutrition 0.000 description 4
- 235000011852 gelatine desserts Nutrition 0.000 description 4
- 239000008187 granular material Substances 0.000 description 4
- 238000009396 hybridization Methods 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 230000006698 induction Effects 0.000 description 4
- 230000007574 infarction Effects 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 239000006166 lysate Substances 0.000 description 4
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 4
- 229940118019 malondialdehyde Drugs 0.000 description 4
- 229920000609 methyl cellulose Polymers 0.000 description 4
- 235000010981 methylcellulose Nutrition 0.000 description 4
- 239000001923 methylcellulose Substances 0.000 description 4
- 238000010172 mouse model Methods 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 4
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 4
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 4
- 238000002203 pretreatment Methods 0.000 description 4
- 230000001681 protective effect Effects 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 3
- 229920000936 Agarose Polymers 0.000 description 3
- 206010003497 Asphyxia Diseases 0.000 description 3
- 238000006418 Brown reaction Methods 0.000 description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 3
- 108020003215 DNA Probes Proteins 0.000 description 3
- 239000003298 DNA probe Substances 0.000 description 3
- 206010012438 Dermatitis atopic Diseases 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 101150074355 GS gene Proteins 0.000 description 3
- 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 3
- 206010019280 Heart failures Diseases 0.000 description 3
- 229920002971 Heparan sulfate Polymers 0.000 description 3
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 3
- 101001076407 Homo sapiens Interleukin-1 receptor antagonist protein Proteins 0.000 description 3
- 101000950847 Homo sapiens Macrophage migration inhibitory factor Proteins 0.000 description 3
- 101001011645 Homo sapiens Muellerian-inhibiting factor Proteins 0.000 description 3
- 102100023530 Interleukin-1 receptor-associated kinase 3 Human genes 0.000 description 3
- 101710199012 Interleukin-1 receptor-associated kinase 3 Proteins 0.000 description 3
- 239000007836 KH2PO4 Substances 0.000 description 3
- 102000019149 MAP kinase activity proteins Human genes 0.000 description 3
- 108040008097 MAP kinase activity proteins Proteins 0.000 description 3
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 3
- 102000003992 Peroxidases Human genes 0.000 description 3
- 239000004698 Polyethylene Substances 0.000 description 3
- 238000002123 RNA extraction Methods 0.000 description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- 240000008042 Zea mays Species 0.000 description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 3
- 210000001015 abdomen Anatomy 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 235000010443 alginic acid Nutrition 0.000 description 3
- 229920000615 alginic acid Polymers 0.000 description 3
- 150000001412 amines Chemical group 0.000 description 3
- 210000000709 aorta Anatomy 0.000 description 3
- 230000006793 arrhythmia Effects 0.000 description 3
- 206010003246 arthritis Diseases 0.000 description 3
- 208000006673 asthma Diseases 0.000 description 3
- 201000008937 atopic dermatitis Diseases 0.000 description 3
- 238000010009 beating Methods 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 235000020958 biotin Nutrition 0.000 description 3
- 239000001110 calcium chloride Substances 0.000 description 3
- 229910001628 calcium chloride Inorganic materials 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 239000001768 carboxy methyl cellulose Substances 0.000 description 3
- 235000010980 cellulose Nutrition 0.000 description 3
- 229920002678 cellulose Polymers 0.000 description 3
- 239000007795 chemical reaction product Substances 0.000 description 3
- 239000007979 citrate buffer Substances 0.000 description 3
- 239000003086 colorant Substances 0.000 description 3
- 235000005822 corn Nutrition 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 230000003292 diminished effect Effects 0.000 description 3
- 210000002615 epidermis Anatomy 0.000 description 3
- 238000013401 experimental design Methods 0.000 description 3
- 210000002950 fibroblast Anatomy 0.000 description 3
- 235000003599 food sweetener Nutrition 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 125000005843 halogen group Chemical group 0.000 description 3
- 210000002064 heart cell Anatomy 0.000 description 3
- 102000057097 human MIF Human genes 0.000 description 3
- 238000003384 imaging method Methods 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 231100000518 lethal Toxicity 0.000 description 3
- 230000001665 lethal effect Effects 0.000 description 3
- 238000011068 loading method Methods 0.000 description 3
- 230000033001 locomotion Effects 0.000 description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 3
- 150000004702 methyl esters Chemical class 0.000 description 3
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 3
- 238000000386 microscopy Methods 0.000 description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 3
- 230000001338 necrotic effect Effects 0.000 description 3
- 230000009871 nonspecific binding Effects 0.000 description 3
- 210000003540 papillary muscle Anatomy 0.000 description 3
- 230000007310 pathophysiology Effects 0.000 description 3
- 108040007629 peroxidase activity proteins Proteins 0.000 description 3
- 230000002085 persistent effect Effects 0.000 description 3
- 239000012660 pharmacological inhibitor Substances 0.000 description 3
- 229920002401 polyacrylamide Polymers 0.000 description 3
- 229920000573 polyethylene Polymers 0.000 description 3
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 3
- 238000002731 protein assay Methods 0.000 description 3
- 238000000751 protein extraction Methods 0.000 description 3
- 230000003134 recirculating effect Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000012552 review Methods 0.000 description 3
- 206010039073 rheumatoid arthritis Diseases 0.000 description 3
- 230000003248 secreting effect Effects 0.000 description 3
- 235000010413 sodium alginate Nutrition 0.000 description 3
- 239000000661 sodium alginate Substances 0.000 description 3
- 229940005550 sodium alginate Drugs 0.000 description 3
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 3
- 238000002798 spectrophotometry method Methods 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 235000000346 sugar Nutrition 0.000 description 3
- 150000008163 sugars Chemical class 0.000 description 3
- 239000003765 sweetening agent Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 229940037128 systemic glucocorticoids Drugs 0.000 description 3
- 239000008399 tap water Substances 0.000 description 3
- 235000020679 tap water Nutrition 0.000 description 3
- 210000001550 testis Anatomy 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- 239000001993 wax Substances 0.000 description 3
- 125000004169 (C1-C6) alkyl group Chemical group 0.000 description 2
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 2
- 101800000733 Angiotensin-2 Proteins 0.000 description 2
- 102400000345 Angiotensin-2 Human genes 0.000 description 2
- 101100339431 Arabidopsis thaliana HMGB2 gene Proteins 0.000 description 2
- 241000416162 Astragalus gummifer Species 0.000 description 2
- 229930003347 Atropine Natural products 0.000 description 2
- 241000972773 Aulopiformes Species 0.000 description 2
- 102100021943 C-C motif chemokine 2 Human genes 0.000 description 2
- 101710155857 C-C motif chemokine 2 Proteins 0.000 description 2
- 102100021824 COP9 signalosome complex subunit 5 Human genes 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 208000020446 Cardiac disease Diseases 0.000 description 2
- 206010007559 Cardiac failure congestive Diseases 0.000 description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 238000011891 EIA kit Methods 0.000 description 2
- 206010018378 Glomerulonephritis rapidly progressive Diseases 0.000 description 2
- 239000007995 HEPES buffer Substances 0.000 description 2
- 108700010013 HMGB1 Proteins 0.000 description 2
- 101150021904 HMGB1 gene Proteins 0.000 description 2
- 208000013875 Heart injury Diseases 0.000 description 2
- 102100037907 High mobility group protein B1 Human genes 0.000 description 2
- 101000896048 Homo sapiens COP9 signalosome complex subunit 5 Proteins 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- RKUNBYITZUJHSG-UHFFFAOYSA-N Hyosciamin-hydrochlorid Natural products CN1C(C2)CCC1CC2OC(=O)C(CO)C1=CC=CC=C1 RKUNBYITZUJHSG-UHFFFAOYSA-N 0.000 description 2
- CZGUSIXMZVURDU-JZXHSEFVSA-N Ile(5)-angiotensin II Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C=CC=CC=1)C([O-])=O)NC(=O)[C@@H](NC(=O)[C@H](CCCNC(N)=[NH2+])NC(=O)[C@@H]([NH3+])CC([O-])=O)C(C)C)C1=CC=C(O)C=C1 CZGUSIXMZVURDU-JZXHSEFVSA-N 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 229940119178 Interleukin 1 receptor antagonist Drugs 0.000 description 2
- 102000051628 Interleukin-1 receptor antagonist Human genes 0.000 description 2
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 206010024119 Left ventricular failure Diseases 0.000 description 2
- 240000007472 Leucaena leucocephala Species 0.000 description 2
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 2
- NNJVILVZKWQKPM-UHFFFAOYSA-N Lidocaine Chemical compound CCN(CC)CC(=O)NC1=C(C)C=CC=C1C NNJVILVZKWQKPM-UHFFFAOYSA-N 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 2
- 101001018878 Mus musculus Macrophage migration inhibitory factor Proteins 0.000 description 2
- 102100029438 Nitric oxide synthase, inducible Human genes 0.000 description 2
- 101710089543 Nitric oxide synthase, inducible Proteins 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 208000002193 Pain Diseases 0.000 description 2
- 239000004792 Prolene Substances 0.000 description 2
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 2
- 241000219061 Rheum Species 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 244000061456 Solanum tuberosum Species 0.000 description 2
- 235000002595 Solanum tuberosum Nutrition 0.000 description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 2
- 239000005864 Sulphur Substances 0.000 description 2
- 229920001615 Tragacanth Polymers 0.000 description 2
- 241000209140 Triticum Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 102000004903 Troponin Human genes 0.000 description 2
- 108090001027 Troponin Proteins 0.000 description 2
- 206010054094 Tumour necrosis Diseases 0.000 description 2
- 206010053613 Type IV hypersensitivity reaction Diseases 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 230000001919 adrenal effect Effects 0.000 description 2
- 230000002411 adverse Effects 0.000 description 2
- 239000000443 aerosol Substances 0.000 description 2
- 239000000783 alginic acid Substances 0.000 description 2
- 229960001126 alginic acid Drugs 0.000 description 2
- 150000004781 alginic acids Chemical class 0.000 description 2
- 125000003545 alkoxy group Chemical group 0.000 description 2
- 239000012491 analyte Substances 0.000 description 2
- 229950006323 angiotensin ii Drugs 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000003042 antagnostic effect Effects 0.000 description 2
- 230000003092 anti-cytokine Effects 0.000 description 2
- 230000000692 anti-sense effect Effects 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 238000009175 antibody therapy Methods 0.000 description 2
- 125000003118 aryl group Chemical group 0.000 description 2
- 210000002072 atrial myocyte Anatomy 0.000 description 2
- RKUNBYITZUJHSG-SPUOUPEWSA-N atropine Chemical compound O([C@H]1C[C@H]2CC[C@@H](C1)N2C)C(=O)C(CO)C1=CC=CC=C1 RKUNBYITZUJHSG-SPUOUPEWSA-N 0.000 description 2
- 229960000396 atropine Drugs 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 230000008827 biological function Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 229940087828 buprenex Drugs 0.000 description 2
- 229960001736 buprenorphine Drugs 0.000 description 2
- CJZGTCYPCWQAJB-UHFFFAOYSA-L calcium stearate Chemical class [Ca+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O CJZGTCYPCWQAJB-UHFFFAOYSA-L 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
- 230000001269 cardiogenic effect Effects 0.000 description 2
- 206010007625 cardiogenic shock Diseases 0.000 description 2
- 230000003293 cardioprotective effect Effects 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 230000005754 cellular signaling Effects 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- AIXMJTYHQHQJLU-UHFFFAOYSA-N chembl210858 Chemical compound O1C(CC(=O)OC)CC(C=2C=CC(O)=CC=2)=N1 AIXMJTYHQHQJLU-UHFFFAOYSA-N 0.000 description 2
- 238000007906 compression Methods 0.000 description 2
- 230000006835 compression Effects 0.000 description 2
- 230000008828 contractile function Effects 0.000 description 2
- 238000013270 controlled release Methods 0.000 description 2
- 201000005637 crescentic glomerulonephritis Diseases 0.000 description 2
- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 description 2
- SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 230000008713 feedback mechanism Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 238000005469 granulation Methods 0.000 description 2
- 230000003179 granulation Effects 0.000 description 2
- 230000005986 heart dysfunction Effects 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 230000000984 immunochemical effect Effects 0.000 description 2
- 229940127121 immunoconjugate Drugs 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- 229940072221 immunoglobulins Drugs 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 208000027866 inflammatory disease Diseases 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 230000003601 intercostal effect Effects 0.000 description 2
- 239000003407 interleukin 1 receptor blocking agent Substances 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 230000031146 intracellular signal transduction Effects 0.000 description 2
- 229960002725 isoflurane Drugs 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 229960004194 lidocaine Drugs 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 235000019359 magnesium stearate Nutrition 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 239000008155 medical solution Substances 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- 239000004570 mortar (masonry) Substances 0.000 description 2
- 210000000440 neutrophil Anatomy 0.000 description 2
- 210000004940 nucleus Anatomy 0.000 description 2
- 230000002018 overexpression Effects 0.000 description 2
- 230000036407 pain Effects 0.000 description 2
- 230000007170 pathology Effects 0.000 description 2
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 2
- 206010034674 peritonitis Diseases 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 230000001817 pituitary effect Effects 0.000 description 2
- 239000000902 placebo Substances 0.000 description 2
- 229940068196 placebo Drugs 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000002685 pulmonary effect Effects 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 239000011535 reaction buffer Substances 0.000 description 2
- 230000009257 reactivity Effects 0.000 description 2
- 210000001567 regular cardiac muscle cell of ventricle Anatomy 0.000 description 2
- 238000010242 retro-orbital bleeding Methods 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 235000019515 salmon Nutrition 0.000 description 2
- 239000012723 sample buffer Substances 0.000 description 2
- 229920006298 saran Polymers 0.000 description 2
- 210000004894 snout Anatomy 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 description 2
- 239000004299 sodium benzoate Substances 0.000 description 2
- 235000010234 sodium benzoate Nutrition 0.000 description 2
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 2
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000012730 sustained-release form Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 239000006188 syrup Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- 239000000454 talc Substances 0.000 description 2
- 229910052623 talc Inorganic materials 0.000 description 2
- 235000012222 talc Nutrition 0.000 description 2
- 210000001541 thymus gland Anatomy 0.000 description 2
- 208000037816 tissue injury Diseases 0.000 description 2
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 2
- 235000010487 tragacanth Nutrition 0.000 description 2
- 239000000196 tragacanth Substances 0.000 description 2
- 229940116362 tragacanth Drugs 0.000 description 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 2
- 230000001052 transient effect Effects 0.000 description 2
- 238000011269 treatment regimen Methods 0.000 description 2
- 230000004565 tumor cell growth Effects 0.000 description 2
- 230000005951 type IV hypersensitivity Effects 0.000 description 2
- 208000027930 type IV hypersensitivity disease Diseases 0.000 description 2
- 238000009423 ventilation Methods 0.000 description 2
- NDQQRRVKUBPTHQ-QBIQUQHTSA-N (2r,3r,4r,5s)-6-(methylamino)hexane-1,2,3,4,5-pentol Chemical compound CNC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO.CNC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO NDQQRRVKUBPTHQ-QBIQUQHTSA-N 0.000 description 1
- LNAZSHAWQACDHT-XIYTZBAFSA-N (2r,3r,4s,5r,6s)-4,5-dimethoxy-2-(methoxymethyl)-3-[(2s,3r,4s,5r,6r)-3,4,5-trimethoxy-6-(methoxymethyl)oxan-2-yl]oxy-6-[(2r,3r,4s,5r,6r)-4,5,6-trimethoxy-2-(methoxymethyl)oxan-3-yl]oxyoxane Chemical compound CO[C@@H]1[C@@H](OC)[C@H](OC)[C@@H](COC)O[C@H]1O[C@H]1[C@H](OC)[C@@H](OC)[C@H](O[C@H]2[C@@H]([C@@H](OC)[C@H](OC)O[C@@H]2COC)OC)O[C@@H]1COC LNAZSHAWQACDHT-XIYTZBAFSA-N 0.000 description 1
- 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 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-M 3-carboxy-2,3-dihydroxypropanoate Chemical compound OC(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-M 0.000 description 1
- BLYOHTUMYRSEOJ-UHFFFAOYSA-N 4-(4,5-dihydro-1,2-oxazol-3-yl)phenol Chemical compound C1=CC(O)=CC=C1C1=NOCC1 BLYOHTUMYRSEOJ-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 208000013824 Acidemia Diseases 0.000 description 1
- 208000010444 Acidosis Diseases 0.000 description 1
- 244000005852 Adiantum capillus veneris Species 0.000 description 1
- 229920001817 Agar Polymers 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
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 206010002383 Angina Pectoris Diseases 0.000 description 1
- 206010003645 Atopy Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 206010070545 Bacterial translocation Diseases 0.000 description 1
- 241001474374 Blennius Species 0.000 description 1
- 229910001369 Brass Inorganic materials 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 1
- MJUYJRCTTJJTDM-UHFFFAOYSA-N CC(C)C(C)(C)C.[V] Chemical compound CC(C)C(C)(C)C.[V] MJUYJRCTTJJTDM-UHFFFAOYSA-N 0.000 description 1
- XAZKFISIRYLAEE-UHFFFAOYSA-N CC1CC(C)CC1 Chemical compound CC1CC(C)CC1 XAZKFISIRYLAEE-UHFFFAOYSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 208000031229 Cardiomyopathies Diseases 0.000 description 1
- 108010078791 Carrier Proteins Proteins 0.000 description 1
- 240000008886 Ceratonia siliqua Species 0.000 description 1
- 235000013912 Ceratonia siliqua Nutrition 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- 241000252203 Clupea harengus Species 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 208000002330 Congenital Heart Defects Diseases 0.000 description 1
- 239000000055 Corticotropin-Releasing Hormone Substances 0.000 description 1
- 244000303965 Cyamopsis psoralioides Species 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- DSLZVSRJTYRBFB-LLEIAEIESA-N D-glucaric acid Chemical compound OC(=O)[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O DSLZVSRJTYRBFB-LLEIAEIESA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- RGHNJXZEOKUKBD-SQOUGZDYSA-M D-gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O RGHNJXZEOKUKBD-SQOUGZDYSA-M 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- 206010052337 Diastolic dysfunction Diseases 0.000 description 1
- 235000019739 Dicalciumphosphate Nutrition 0.000 description 1
- JRWZLRBJNMZMFE-UHFFFAOYSA-N Dobutamine Chemical compound C=1C=C(O)C(O)=CC=1CCNC(C)CCC1=CC=C(O)C=C1 JRWZLRBJNMZMFE-UHFFFAOYSA-N 0.000 description 1
- 102400000686 Endothelin-1 Human genes 0.000 description 1
- 101800004490 Endothelin-1 Proteins 0.000 description 1
- 108010008165 Etanercept Proteins 0.000 description 1
- 208000009386 Experimental Arthritis Diseases 0.000 description 1
- 102000007665 Extracellular Signal-Regulated MAP Kinases Human genes 0.000 description 1
- 108010007457 Extracellular Signal-Regulated MAP Kinases Proteins 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 1
- 229940123037 Glucocorticoid antagonist Drugs 0.000 description 1
- 229920002907 Guar gum Polymers 0.000 description 1
- 208000035211 Heart Murmurs Diseases 0.000 description 1
- 101000669447 Homo sapiens Toll-like receptor 4 Proteins 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 206010020880 Hypertrophy Diseases 0.000 description 1
- 206010058558 Hypoperfusion Diseases 0.000 description 1
- 108010021699 I-kappa B Proteins Proteins 0.000 description 1
- 102000008379 I-kappa B Proteins Human genes 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 1
- 108010054477 Immunoglobulin Fab Fragments Proteins 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
- 108010050904 Interferons Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 102100036342 Interleukin-1 receptor-associated kinase 1 Human genes 0.000 description 1
- 101710199015 Interleukin-1 receptor-associated kinase 1 Proteins 0.000 description 1
- 206010022680 Intestinal ischaemia Diseases 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-L L-tartrate(2-) Chemical compound [O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O FEWJPZIEWOKRBE-JCYAYHJZSA-L 0.000 description 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 description 1
- 239000012741 Laemmli sample buffer Substances 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Chemical compound CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 208000019693 Lung disease Diseases 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 102000043131 MHC class II family Human genes 0.000 description 1
- 108091054438 MHC class II family Proteins 0.000 description 1
- 101150058224 MIF gene Proteins 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 229920000881 Modified starch Polymers 0.000 description 1
- 229930191564 Monensin Natural products 0.000 description 1
- GAOZTHIDHYLHMS-UHFFFAOYSA-N Monensin A Natural products O1C(CC)(C2C(CC(O2)C2C(CC(C)C(O)(CO)O2)C)C)CCC1C(O1)(C)CCC21CC(O)C(C)C(C(C)C(OC)C(C)C(O)=O)O2 GAOZTHIDHYLHMS-UHFFFAOYSA-N 0.000 description 1
- 101001082628 Mus musculus H-2 class II histocompatibility antigen gamma chain Proteins 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 206010033557 Palpitations Diseases 0.000 description 1
- 229920001774 Perfluoroether Chemical group 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 102000003923 Protein Kinase C Human genes 0.000 description 1
- 108090000315 Protein Kinase C Proteins 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 241000219492 Quercus Species 0.000 description 1
- 206010040893 Skin necrosis Diseases 0.000 description 1
- BCKXLBQYZLBQEK-KVVVOXFISA-M Sodium oleate Chemical compound [Na+].CCCCCCCC\C=C/CCCCCCCC([O-])=O BCKXLBQYZLBQEK-KVVVOXFISA-M 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- 240000001058 Sterculia urens Species 0.000 description 1
- 235000015125 Sterculia urens Nutrition 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 206010071436 Systolic dysfunction Diseases 0.000 description 1
- 239000008049 TAE buffer Substances 0.000 description 1
- 102100039360 Toll-like receptor 4 Human genes 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 108010078814 Tumor Suppressor Protein p53 Proteins 0.000 description 1
- 102000015098 Tumor Suppressor Protein p53 Human genes 0.000 description 1
- 102100033732 Tumor necrosis factor receptor superfamily member 1A Human genes 0.000 description 1
- 101710187743 Tumor necrosis factor receptor superfamily member 1A Proteins 0.000 description 1
- 108010000134 Vascular Cell Adhesion Molecule-1 Proteins 0.000 description 1
- 102100023543 Vascular cell adhesion protein 1 Human genes 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- HGEVZDLYZYVYHD-UHFFFAOYSA-N acetic acid;2-amino-2-(hydroxymethyl)propane-1,3-diol;2-[2-[bis(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetic acid Chemical compound CC(O)=O.OCC(N)(CO)CO.OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O HGEVZDLYZYVYHD-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 206010069351 acute lung injury Diseases 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 238000013228 adult male C57BL/6J mice Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000010419 agar Nutrition 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 1
- 125000003342 alkenyl group Chemical group 0.000 description 1
- 125000004453 alkoxycarbonyl group Chemical group 0.000 description 1
- 125000004457 alkyl amino carbonyl group Chemical group 0.000 description 1
- 125000003282 alkyl amino group Chemical group 0.000 description 1
- 125000004448 alkyl carbonyl group Chemical group 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 125000004390 alkyl sulfonyl group Chemical group 0.000 description 1
- 125000004414 alkyl thio group Chemical group 0.000 description 1
- 125000000304 alkynyl group Chemical group 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- SNAAJJQQZSMGQD-UHFFFAOYSA-N aluminum magnesium Chemical compound [Mg].[Al] SNAAJJQQZSMGQD-UHFFFAOYSA-N 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 238000002399 angioplasty Methods 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000001387 anti-histamine Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000003416 antiarrhythmic agent Substances 0.000 description 1
- 210000000612 antigen-presenting cell Anatomy 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 125000003710 aryl alkyl group Chemical group 0.000 description 1
- 125000005129 aryl carbonyl group Chemical group 0.000 description 1
- 125000005161 aryl oxy carbonyl group Chemical group 0.000 description 1
- 125000004391 aryl sulfonyl group Chemical group 0.000 description 1
- 125000004104 aryloxy group Chemical group 0.000 description 1
- 210000003567 ascitic fluid Anatomy 0.000 description 1
- 230000003190 augmentative effect Effects 0.000 description 1
- 230000003305 autocrine Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000007375 bacterial translocation Effects 0.000 description 1
- 235000012216 bentonite Nutrition 0.000 description 1
- SRSXLGNVWSONIS-UHFFFAOYSA-M benzenesulfonate Chemical compound [O-]S(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-M 0.000 description 1
- 229940077388 benzenesulfonate Drugs 0.000 description 1
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 235000010338 boric acid Nutrition 0.000 description 1
- 239000010951 brass Substances 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000004067 bulking agent Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 235000010410 calcium alginate Nutrition 0.000 description 1
- 239000000648 calcium alginate Substances 0.000 description 1
- 229960002681 calcium alginate Drugs 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 235000013539 calcium stearate Nutrition 0.000 description 1
- 239000008116 calcium stearate Substances 0.000 description 1
- 235000011132 calcium sulphate Nutrition 0.000 description 1
- DIOLOCSXUMYFJN-UHFFFAOYSA-N calcium;azane Chemical compound N.[Ca+2] DIOLOCSXUMYFJN-UHFFFAOYSA-N 0.000 description 1
- BPKIGYQJPYCAOW-FFJTTWKXSA-I calcium;potassium;disodium;(2s)-2-hydroxypropanoate;dichloride;dihydroxide;hydrate Chemical compound O.[OH-].[OH-].[Na+].[Na+].[Cl-].[Cl-].[K+].[Ca+2].C[C@H](O)C([O-])=O BPKIGYQJPYCAOW-FFJTTWKXSA-I 0.000 description 1
- 125000003917 carbamoyl group Chemical group [H]N([H])C(*)=O 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 229940105329 carboxymethylcellulose Drugs 0.000 description 1
- 206010061592 cardiac fibrillation Diseases 0.000 description 1
- 230000001848 cardiodepressant effect Effects 0.000 description 1
- 230000002612 cardiopulmonary effect Effects 0.000 description 1
- 239000012876 carrier material Substances 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 230000003081 coactivator Effects 0.000 description 1
- 229940110456 cocoa butter Drugs 0.000 description 1
- 235000019868 cocoa butter Nutrition 0.000 description 1
- 206010009887 colitis Diseases 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000005056 compaction Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- IDLFZVILOHSSID-OVLDLUHVSA-N corticotropin Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)NC(=O)[C@@H](N)CO)C1=CC=C(O)C=C1 IDLFZVILOHSSID-OVLDLUHVSA-N 0.000 description 1
- 229960000258 corticotropin Drugs 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 235000010947 crosslinked sodium carboxy methyl cellulose Nutrition 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000016396 cytokine production Effects 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002939 deleterious effect Effects 0.000 description 1
- KXGVEGMKQFWNSR-LLQZFEROSA-N deoxycholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 KXGVEGMKQFWNSR-LLQZFEROSA-N 0.000 description 1
- 229960003964 deoxycholic acid Drugs 0.000 description 1
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 description 1
- 229960000633 dextran sulfate Drugs 0.000 description 1
- 125000004663 dialkyl amino group Chemical group 0.000 description 1
- 125000004473 dialkylaminocarbonyl group Chemical group 0.000 description 1
- NEFBYIFKOOEVPA-UHFFFAOYSA-K dicalcium phosphate Chemical compound [Ca+2].[Ca+2].[O-]P([O-])([O-])=O NEFBYIFKOOEVPA-UHFFFAOYSA-K 0.000 description 1
- 229940038472 dicalcium phosphate Drugs 0.000 description 1
- 229910000390 dicalcium phosphate Inorganic materials 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 229960001089 dobutamine Drugs 0.000 description 1
- 239000008298 dragée Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000009510 drug design Methods 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 238000007908 dry granulation Methods 0.000 description 1
- 230000002828 effect on organs or tissue Effects 0.000 description 1
- 229940073621 enbrel Drugs 0.000 description 1
- 206010014665 endocarditis Diseases 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 210000003890 endocrine cell Anatomy 0.000 description 1
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 1
- 210000003989 endothelium vascular Anatomy 0.000 description 1
- 210000003979 eosinophil Anatomy 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- CCIVGXIOQKPBKL-UHFFFAOYSA-M ethanesulfonate Chemical compound CCS([O-])(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-M 0.000 description 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 239000002095 exotoxin Substances 0.000 description 1
- 231100000776 exotoxin Toxicity 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 230000002600 fibrillogenic effect Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000003205 genotyping method Methods 0.000 description 1
- 229960004580 glibenclamide Drugs 0.000 description 1
- 239000003635 glucocorticoid antagonist Substances 0.000 description 1
- 229940050410 gluconate Drugs 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 229930195712 glutamate Natural products 0.000 description 1
- ZNNLBTZKUZBEKO-UHFFFAOYSA-N glyburide Chemical compound COC1=CC=C(Cl)C=C1C(=O)NCCC1=CC=C(S(=O)(=O)NC(=O)NC2CCCCC2)C=C1 ZNNLBTZKUZBEKO-UHFFFAOYSA-N 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 235000010417 guar gum Nutrition 0.000 description 1
- 239000000665 guar gum Substances 0.000 description 1
- 229960002154 guar gum Drugs 0.000 description 1
- 210000004837 gut-associated lymphoid tissue Anatomy 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- 150000002367 halogens Chemical class 0.000 description 1
- 239000007887 hard shell capsule Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000000004 hemodynamic effect Effects 0.000 description 1
- 235000019514 herring Nutrition 0.000 description 1
- 125000004475 heteroaralkyl group Chemical group 0.000 description 1
- 125000001072 heteroaryl group Chemical group 0.000 description 1
- 125000005553 heteroaryloxy group Chemical group 0.000 description 1
- 239000008240 homogeneous mixture Substances 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 102000046824 human IL1RN Human genes 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-M hydrogensulfate Chemical compound OS([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-M 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- 239000001866 hydroxypropyl methyl cellulose Substances 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
- 230000028993 immune response Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000013388 immunohistochemistry analysis Methods 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 239000003999 initiator Substances 0.000 description 1
- 230000015788 innate immune response Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 230000003870 intestinal permeability Effects 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
- 230000001788 irregular Effects 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 229940116871 l-lactate Drugs 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000007942 layered tablet Substances 0.000 description 1
- 231100000636 lethal dose Toxicity 0.000 description 1
- 231100000225 lethality Toxicity 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 1
- 201000005249 lung adenocarcinoma Diseases 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 238000013227 male C57BL/6J mice Methods 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 102000006240 membrane receptors Human genes 0.000 description 1
- 108020004084 membrane receptors Proteins 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000002493 microarray Methods 0.000 description 1
- 238000010208 microarray analysis Methods 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- ZAHQPTJLOCWVPG-UHFFFAOYSA-N mitoxantrone dihydrochloride Chemical compound Cl.Cl.O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO ZAHQPTJLOCWVPG-UHFFFAOYSA-N 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 235000019426 modified starch Nutrition 0.000 description 1
- 229960005358 monensin Drugs 0.000 description 1
- GAOZTHIDHYLHMS-KEOBGNEYSA-N monensin A Chemical compound C([C@@](O1)(C)[C@H]2CC[C@@](O2)(CC)[C@H]2[C@H](C[C@@H](O2)[C@@H]2[C@H](C[C@@H](C)[C@](O)(CO)O2)C)C)C[C@@]21C[C@H](O)[C@@H](C)[C@@H]([C@@H](C)[C@@H](OC)[C@H](C)C(O)=O)O2 GAOZTHIDHYLHMS-KEOBGNEYSA-N 0.000 description 1
- 238000002625 monoclonal antibody therapy Methods 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 210000002200 mouth mucosa Anatomy 0.000 description 1
- 230000005914 myocardial expression Effects 0.000 description 1
- 229920001206 natural gum Polymers 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 238000006384 oligomerization reaction Methods 0.000 description 1
- 239000003605 opacifier Substances 0.000 description 1
- 239000007935 oral tablet Substances 0.000 description 1
- 125000004043 oxo group Chemical group O=* 0.000 description 1
- TWNQGVIAIRXVLR-UHFFFAOYSA-N oxo(oxoalumanyloxy)alumane Chemical compound O=[Al]O[Al]=O TWNQGVIAIRXVLR-UHFFFAOYSA-N 0.000 description 1
- WLJNZVDCPSBLRP-UHFFFAOYSA-N pamoic acid Chemical compound C1=CC=C2C(CC=3C4=CC=CC=C4C=C(C=3O)C(=O)O)=C(O)C(C(O)=O)=CC2=C1 WLJNZVDCPSBLRP-UHFFFAOYSA-N 0.000 description 1
- 230000003076 paracrine Effects 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 125000005010 perfluoroalkyl group Chemical group 0.000 description 1
- 208000008494 pericarditis Diseases 0.000 description 1
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000004014 plasticizer Substances 0.000 description 1
- 229920002451 polyvinyl alcohol Polymers 0.000 description 1
- 235000019422 polyvinyl alcohol Nutrition 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 235000015277 pork Nutrition 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229940114930 potassium stearate Drugs 0.000 description 1
- ANBFRLKBEIFNQU-UHFFFAOYSA-M potassium;octadecanoate Chemical compound [K+].CCCCCCCCCCCCCCCCCC([O-])=O ANBFRLKBEIFNQU-UHFFFAOYSA-M 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- ALDITMKAAPLVJK-UHFFFAOYSA-N prop-1-ene;hydrate Chemical group O.CC=C ALDITMKAAPLVJK-UHFFFAOYSA-N 0.000 description 1
- DNIAPMSPPWPWGF-UHFFFAOYSA-N propylene glycol Substances CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- 201000005825 prostate adenocarcinoma Diseases 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000010410 reperfusion Effects 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 230000020874 response to hypoxia Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 208000004124 rheumatic heart disease Diseases 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 239000007909 solid dosage form Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000012453 sprague-dawley rat model Methods 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000011410 subtraction method Methods 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 1
- 230000008093 supporting effect Effects 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 229940095064 tartrate Drugs 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 230000005945 translocation Effects 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 239000013638 trimer Substances 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 229960001005 tuberculin Drugs 0.000 description 1
- 230000005747 tumor angiogenesis Effects 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 238000005550 wet granulation Methods 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2878—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 NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/06—Antiarrhythmics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- 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
- C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
- C07K16/2833—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against MHC-molecules, e.g. HLA-molecules
-
- 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
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
- A61K2039/507—Comprising a combination of two or more separate antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
Definitions
- the invention generally relates to pathology and physiology in vertebrate species involving cytokines and other cellular signaling mechanisms, and also diagnostic assays involving cytokines and other cellular signaling mechanisms.
- Other aspects of the invention relate to macrophage migration inhibitory factor (MIF) as a myocardial depressant factor and as a mediator of endotoxin-induced cardiac dysfunction in vivo.
- MIF macrophage migration inhibitory factor
- Other aspects of the invention relate to mediating and/or inhibiting the production or activity of MIF, and compounds, compositions, methods of treating and preventing cardiac dysfunction, sepsis, burn injury or other conditions related to burns.
- Other aspects of the invention relate to the MIF release from the heart, liver, and spleen and the role of TNF receptor I/II signaling after LPS challenge.
- Other aspects of the invention relate to TNF receptor I/II signaling independent release of MIF into the serum.
- Other aspects of the invention relate to the expression of CD74 on cardiomyocyte
- Macrophage migration inhibitory factor is a pluripotent, pro-inflammatory cytokine whose mechanisms of action have been scrutinized over the past four decades.
- the current understanding in the art relating to MIF includes studies directed to its crystallization as a trimer, its physiologically relevant oligomerization state; its putative membrane receptor(s); and the physiologic relevance of its intracellular enzymatic activity as a tautomerase and oxidoreductase.
- MIF has an important role in diseases as diverse as rheumatoid arthritis
- M. Leech, et al. “Macrophage Migration Inhibitory Factor in Rheumatoid Arthritis: Evidence of Proinflammatory Function and Regulation by Glucocorticoids”, Arthritis Rheum, 42, 1601-1608 (1999)
- M. Leech, et al. “Involvement of Macrophage Migration Inhibitory Factor in the Evolution of Rat Adjuvant Arthritis”, Arthritis Rheum., 41, 910-917 (1998), A.
- MIF is a Pituitary-Derived Cytokine that Potentiates Lethal Endotoxaemia”, Nature, 365, 756-759 (1993), M. Bozza, et al., “Targeted Disruption of Migration Inhibitory Factor Gene Reveals Its Critical Role in Sepsis”, J. Exp. Med. 189, 341-346 (1999), T. Calandra, et al., “MIF as a Glucocorticoid-Induced Modulator of Cytokine Production”, Nature, 377, 68-71 (1995), T.
- MIF is increased in the plasma of animals and humans, and the blockade of MIF activity by monoclonal or polyclonal antibodies results in a marked improvement in the survival of animals with experimentally induced sepsis (M. Bozza, et al., “Targeted Disruption of Migration Inhibitory Factor Gene Reveals Its Critical Role in Sepsis”, J. Exp. Med. 189, 341-346 (1999), T. Calandra, et al., “Protection from Septic Shock by Neutralization of Macrophage Migration Inhibitory Factor”, Nat. Med., 6, 164-170 (2000)).
- Blockade of MIF activity has been demonstrated with a number of inhibitors.
- Blockade of MIF enzymatic activity has been demonstrated with diverse chemical compounds as shown in U.S. patent application Ser. No. 10/226,299, filed Aug. 23, 2002, now pending. See also, for instance, U.S. Pat. No. 6,492,428.
- Antibodies have also been used to blockade MIF activity as shown in U.S. Pat. No. 6,030,615.
- MIF expression can also be inhibited using antisense technology as disclosed in U.S. patent application Ser. No. 08/738,947, filed Oct. 24, 1996, now pending, or U.S. Pat. No. 6,268,151 which further demonstrates pharmaceutical formulations that can be used with all the above-mentioned MIF inhibitors.
- LPS Lipopolysaccharide depresses intrinsic myocardial contractility and is thought to be important in myocardial dysfunction that occurs in sepsis and septic shock
- A. M. Lefer “Mechanisms of cardiodepression in endotoxin shock”, Circ Shock Suppl 1:1-8 (1979), J. E. Parrillo, et al., “A circulating myocardial depressant substance in humans with septic shock. Septic shock patients with a reduced ejection fraction have a circulating factor that depresses in vitro myocartdial cell performance”, J. Clin. Invest. 76:1539-1553 (1985), J. M.
- Macrophage migration inhibitory factor is a caridac-derived myocardial depressant factor
- S. Krishnagopalan, et al. “Myocardial dysfunction in the patient with sepsis”, Curr. Opin. Crit. Care 8:376-388 (2002)).
- MIF macrophage migration inhibitory factor
- Macrophage migration inhibitory factor is involved in the pathogenesis of several diseases, including sepsis. MIF opposes the anti-inflammatory effects of glucocorticoids, and also significantly alters tissue metabolism. Although MIF appears to be ubiquitously expressed, there are currently no publications indicating whether MIF is expressed in the myocardium in vivo, or whether release of MIF from the myocardium or other tissues during sepsis could adversely affect cardiac performance.
- mice were protected when the antibodies were given as late as 8 h after the onset of infection (T. Calandra, et al., “Protection from septic shock by neutralization of macrophage migration inhibitory factor”, Nat. Med., 6, 164-170 (2000)).
- MIF has a number of properties that make it unique among cytokines. MIF is released preformed from numerous cell types including lymphocytes, macrophages, and the anterior pituitary (J. Bernhagen, et al., “Regulation of the Immune Response by Macrophage Migration Inhibitory Factor: Biological and Structural Features”, J. Mol. Med., 76, 151-161 (1998), T. Calandra, et al., “Macrophage Migration Inhibitory Factor (MIF): A Glucocorticoid Counter-Regulator Within the Immune System”, Crit. Rev. Immunol., 17, 77-88 (1997), S. C.
- MIF Macrophage Migration Inhibitory Factor
- MIF has at least 2 catalytic activities that are distinct: tautomerase and oxidoreductase activity.
- MIF-related diseases such as sepsis, acute respiratory distress syndrome (ARDS), asthma, atopic dermatitis, rheumatoid arthritis, nephropathy, and cancer
- ARDS acute respiratory distress syndrome
- asthma asthma
- atopic dermatitis rheumatoid arthritis
- nephropathy nephropathy
- cancer nephropathy
- MIF Macrophages expressing anti-sense MIF cDNA (leading to less endogenous MIF) secrete/express significantly less TNF- ⁇ , IL-6, and NO, while NF- ⁇ B activity is decreased in response to LPS (44). Therefore, it appears that MIF may directly interact with the LPS signaling pathway (H. Lue, et al., “Macrophage Migration Inhibitory Factor (MIF): Mechanisms of Action and Role in Disease”, Microbes Infect., 4, 449-460 (2002)).
- MIF Macrophage Migration Inhibitory Factor
- MIF knockout mice as demonstrated in U.S. patent application Ser. No. ______ (Attorney Docket No. 9551-095-27) filed Dec. 19, 2002, which are resistant to lethal doses of LPS, have lower circulating plasma levels of TNF- ⁇ compared to wild-type mice at baseline.
- LPS challenge Upon LPS challenge, they demonstrate diminished circulating TNF- ⁇ concentrations, increased nitric oxide (NO) concentrations, and unchanged IL-6 and IL-12 concentrations (M. Bozza, et al., “Targeted Disruption of Migration Inhibitory Factor Gene Reveals Its Critical Role in Sepsis”, J. Exp. Med. 189, 341-346 (1999)).
- MIF appears to promote pro-inflammatory cytokines
- the effects of MIF have been shown to act in a TNF- ⁇ -independent manner.
- CLP was performed in TNF- ⁇ knock out mice
- a 60% survival rate was seen in mice administered anti-MIF antibodies compared to a 0% survival rate in wild-type mice (T. Calandra, et al., “Protection from Septic Shock by Neutralization of Macrophage Migration Inhibitory Factor”, Nat. Med., 6, 164-170 (2000)).
- Such dysfunctions include, but are not limited to, mycarditis, endocarditis, pericarditis, rheumatic heart disease, myocardial infarction, arrythmia, fibrillation, cardiogenic shock, ischemia, hypertrophy, cardiomyopathy, angina, heart murmur or palpitation, heart attack or failure, and any of the symptoms or defects associated with congenital heart diseases generally.
- Macrophage migration inhibitory factor is a expressed in many organs including the heart and has been linked with a delayed cardiac dysfunction in a murine model of endotoxicosis (Garner, et al., “Macrophage Migration Inhibitory Factor is A Cardiac-Derived Myocardial Depressant Factor”, Am. J. Physiol. Heart Circ. Physiol, 258, H2500-H2509 (2003)).
- Burn injury results in cardiac injury and contractile dysfunction involving decreased cardiac output, shock, and left ventricular failure (J. T. Murphy, et al., “Evaluation of Troponin-I as An Indicator of Cardiac Dysfunction After Thermal Injury, 45, 700-704, (1998), E. M. Reynolds, et al., “Left Ventricular Failure Complicating Severe Pediatric Burn Injuries”, J. Pediatr. Surg. 30, 264-269; discussion 269-270 (1995), W. C. Shoemaker, et al., k′′Burn Pathophysiology In Man. I. Sequential Hemodynamic Alterations, J. Surg. Res., 14, 64-73 (1973), R. R.
- MIF macrophage migration inhibitory factor
- MIF has been shown to play a key role in ARDS (K. N. Lai, et al., “Role For Macrophage Migration Inhibitory Factor in Acute Respiratory Distress Syndrome”, J. Pathol., 199, 496-508 (2003)), a common complication of burn injury (M. Bhatia, et al., “Role of Inflammatory Mediators in the Pathophysiology of Acute Respiratory Distress Syndrome”, J. Pathol., 202, 145-156 (2004)).
- U.S. Pat. No. 6,030,615 relates to methods and compositions for treating a disease caused by cytokine-mediated toxicity.
- U.S. Pat. No. 6,420,188 relates to methods and compositions for antagonizing MIF activity and methods of treating various diseases based on this activity.
- U.S. Pat. No. 6,599,938 relates to methods and compositions for antagonizing MIF activity and methods of treating various diseases based on this activity.
- U.S. Pat. No. 6,645,493 relates to compositions and methods for inhibiting the release and/or biological activity of MIF.
- One embodiment of the present invention relates to a pharmaceutical composition effective for at least one selected from the group including treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, and a combination thereof, which includes:
- Another embodiment of the present invention relates to a pharmaceutical composition effective for at least one selected from the group including treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, and a combination thereof, which includes:
- Another embodiment of the present invention relates to a pharmaceutical composition effective for at least one selected from the group including treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, and a combination thereof, which includes:
- Another embodiment of the present invention relates to a method for treating and/or preventing cardiac dysfunction in a subject, the method including:
- Another embodiment of the present invention relates to a method for treating and/or preventing burn injury-associated cardiac dysfunction in a subject, the method including:
- Another embodiment of the present invention relates to a method for treating and/or preventing cardiac dysfunction in a subject, the method including:
- Another embodiment of the present invention relates to a method for improving cardiac function in a subject following acute myocardial infarction, the method including:
- Another embodiment of the present invention relates to a method for identifying an MIF inhibitor, the method including:
- Another embodiment of the present invention relates to a method for treating and/or preventing cardiac dysfunction in a subject following acute myocardial infarction, the method including:
- Another embodiment of the present invention relates to a method for treating and/or preventing cardiac dysfunction in a subject, the method including:
- Another embodiment of the present invention relates to a method for treating and/or preventing burn injury-associated cardiac dysfunction in a subject, the method including:
- Another embodiment of the present invention relates to a method for treating and/or preventing cardiac dysfunction in a subject, the method including:
- Another embodiment of the present invention relates to a method for improving cardiac function in a subject following acute myocardial infarction, the method including:
- Another embodiment of the present invention relates to a method for treating and/or preventing cardiac dysfunction in a subject following acute myocardial infarction, the method including:
- Another embodiment of the invention relates to a method for at least one selected from the group including treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, and a combination thereof, which includes administering to said subject an effective amount of at least one selected from the group including a small molecule MIF inhibitor, salt thereof, prodrug thereof, and a combination thereof.
- Another embodiment of the invention relates to a method for at least one selected from the group including treating or preventing cardiac dysfunction in a subject in need thereof, treating or preventing irregularity in myocardial activity in a subject in need thereof, treating or preventing depression in myocardial activity in a subject in need thereof, treating or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating or preventing cardiodepression in a subject in need thereof, and a combination thereof, which includes administering to a subject in need thereof an effective amount of at least one anti-TNFR antibody; and
- FIG. 1 MIF protein release is detected within 12 h of LPS challenge in cardiac tissue. Each data point is the mean (+/ ⁇ standard error) of 3 independent Western blot experiments. A representative Western blot is shown below the graph. *p ⁇ 0.05
- FIG. 2 LPS challenge does not upregulate MIF mRNA in cardiac tissue.
- Each data point in the graph is the mean (+/ ⁇ standard error) of 3 independent Northern blot experiments. A representative Northern blot is shown below the graph. No significant differences between time points were identified (p>0.05).
- FIG. 3 The presence of MIF in the heart, liver, and spleen before and after LPS challenge.
- Preformed MIF in the heart, liver, and spleen decreases 12 h after LPS challenge (B, E, and H) and is replenished after 24 h (C, F, and 1) as demonstrated by immunohistochemistry.
- FIG. 4 Cardiac function determined by Langendorff preparation post-rMIF challenge in C57BL/6J mice and endotoxin-resistant C3H/HeJ mice demonstrates rMIF mediates cardiac dysfunction in an LPS-independent mechanism. Data represents the average of 7 (C3H/HeJ) to 10 (C57BL/6J) independent Langendorff experiments per group. *p ⁇ 0.05.
- FIG. 5 Echocardiographic assessment of the effects of LPS and LPS plus anti-MIF antibody administration on cardiac function. Representative M-mode echocardiograms in wild-type mice at baseline and 8 h after LPS administration, A and B, respectively. C and D show, respectively, representative echocardiograms in LPS plus anti-MIF treated mice at 8 and 48 h. A significant protection in cardiac function (FS %) is observed in LPS challenged mice when anti-MIF anti-bodies are given pre-treatment (E). Data represents the average of 9 cardiac cycles from 3 mice monitored at multiple time points. *p ⁇ 0.05.
- FIG. 6 Burn model demonstrating inhibition of MIF with anti-MIF antibody following LPS Challenge.
- the burn data demonstrates inhibition of MIF with the anti-MIF antibody and restores cardiac function following burning.
- FIG. 7 A graphical representation demonstrating MIF release rate from the heart following thermal trauma. Macrophage migration inhibitory factor (MIF) is constitutively expressed in cardiac tissue and released maximally 8 hours post-burn injury. Each data point represents the mean density in arbitrary units (A.U.)/mm 2 ⁇ SE of 3 independent Western blot experiments. A representative Western blot is shown below the graph. A One Way ANOVA and a multiple comparison procedure using the Tukey method were employed to determine statistical significance compared to the control group (*p ⁇ 0.05).
- MIF Macrophage migration inhibitory factor
- FIG. 8 Immunochemistry staining slides demonstrating MIF presence in various tissue samples. Insert G-MIF, constitutively present in the heart, liver, spleen, lung, and kidney is decreased after burn injury. Preformed MIF in the heart, kidney, and spleen (A, E, 1) decreases 8 hours after burn injury (B, F, J), except for liver (M,N) and increases at 24 hours in heart and kidney (C, G), but not heart or liver (K,O) as demonstrated by immunohistochemistry.
- a negative control secondary antibody without the primary anti-MIF antibody
- FIG. 9 Graphical representations of concentration change of three different cytokines over time following thermal trauma.
- FIG. 9 -Serum concentrations of MIF (ng/ml), IL-12 (pg/ml), and IL-6 (pg/ml) following burn injury (A-C, respectively).
- Data are expressed as the mean ⁇ SE of six C57BL/6J mice as determined by ELISA and were statistically analyzed using a One Way-ANOVA with a multiple comparison procedure employing the Bonferroni method to determine significance between groups (*p ⁇ 0.05 compared to baseline).
- FIG. 10 Graphical representation and representative Northern Blot showing MIF mRNA upregulation following thermal trauma. Burn injury upregulates MIF mRNA expression in cardiac tissue significantly by 8 hours. MIF and ⁇ -actin mRNAs were detected using 32 P radiolabeled probes complementary to MIF and ⁇ -actin mRNAs. Each data point represents the mean density in arbitrary units (A.U.)/mm 2 ⁇ SE of 3 independent Northern blot experiments. A representative Northern blot is shown below the graph. Normalized MIF was determined by multiplying the MIF density by the relative ⁇ -actin density present. A One Way-ANOVA and a multiple comparison procedure using the Tukey method were employed to determine statistical significance compared to baseline (*p ⁇ 0.05).
- FIG. 11 Graphical representations of Coronary Flow Rates calculated three different ways comparing control measure of flow rate with untreated thermal trauma versus thermal trauma treated with Anti-MIF. Cardiac function determination by Langendorff preparation 18 hours after burn injury as a function of coronary flow (A) and Ca 2+ . Cardiac function is expressed as the mean ⁇ SE of 25 independent Langendorff experiments (11 sham, 9 burn injury, 5 burn injury after anti-MIF treatment). Separate analyses were performed for each LVP, +dP/dt max , and ⁇ dP/dt max as a function of treatment group and coronary flow rate using a Repeated Measures ANOVA and a multiple comparison procedure employing the Bonferroni method to determine significant differences between groups (*p ⁇ 0.05).
- FIG. 12 Echocardiographic and graphical representation of the effects of anti-MIF antibody therapy after burn injury demonstrating the cardioprotective effects of MIF blockade. Echocardiographic assessment of the effects of anti-MIF antibody therapy after burn injury demonstrates the cardioprotective effects of MIF blockade. Representative M-mode echocardiograms in wild-type mice at baseline and 8 hours after burn injury, A and B, respectively. C and D depict representative echocardiograms in burn injury plus anti-MIF treated mice at 4 and 48 hours, respectively. A significant recovery of cardiac function (FS %) is observed in burn injury mice given anti-MIF anti-bodies pre-burn injury (E). Data from each group represent the mean ⁇ SE of 9 cardiac cycles from 3 mice monitored at multiple time points.
- Cardiac function determined by echocardiography is expressed as fractional shortening % (LVED ⁇ LVES/LVED ⁇ 100) ⁇ SE and was analyzed using a One Way Repeated Measures-ANOVA and a multiple comparison procedure employing the Tukey Test to determine significant differences between specific groups.
- FIG. 13 Serum MIF concentration (fold increase from baseline) following a 4 mg/kg endotoxin challenge in: (A) wild type mice, (B) TNFR ⁇ / ⁇ mice, and (C) wild type mice pre-treated (60 minutes) with Enbrel®. Data are expressed a fraction of the baseline levels of MIF (mean+/ ⁇ standard error) of: (A) 6 C57BL/6J mice, (B) 6 TNFR ⁇ / ⁇ mice, and (C) 3 C57BL/6J (C) pre-treated with Enbrel®. Serum levels were determined by ELISA and were statistically analyzed using a One Way-ANOVA with a multiple comparison procedure employing the Bonferroni method to determine significance between groups (*p ⁇ 0.05 compared to baseline).
- FIG. 14 MIF protein release is not detected after LPS challenge in cardiac tissue. Each data point is the mean (+/ ⁇ standard error) of 3 independent Western blot experiments. A representative Western blot is shown below the graph. *p ⁇ 0.05
- FIG. 15 The presence of MIF in the heart, liver, and spleen before and after LPS challenge in TNFR ⁇ / ⁇ mice.
- Preformed MIF in the heart, liver, and spleen (A, D, G) does not decrease 12 hours after LPS challenge (B, F, J) which is seen in wild type mice as demonstrated by immunohistochemistry.
- FIG. 16 LPS challenge does not upregulate MIF mRNA in cardiac tissue in TNFR ⁇ / ⁇ mice.
- Each data point in the graph is the mean (+/ ⁇ standard error) of 3 independent Northern blot experiments. A representative Northern blot is shown below the graph. No significant differences between time points were identified (p>0.05).
- FIG. 17 Cardiac function determined by Langendorff preparation post-rMIF challenge in C57BL/6J, B6/129S, and TNFR ⁇ / ⁇ mice demonstrates rMIF mediates cardiac dysfunction independent of TNF- ⁇ signaling ex vivo. Data represents the average of 6 (C57BL/6J), 4 (B6/129S), and 4 (TNFR ⁇ / ⁇ ) independent Langendorff experiments per group. *p ⁇ 0.05.
- FIG. 18 Echocardiographic assessment of the effects of LPS and LPS plus anti-MIF antibody administration on cardiac function in TNFR ⁇ / ⁇ mice. Representative M-mode echocardiograms in wild-type mice at baseline and 4 h after LPS administration, A and B, respectively. C and D show, respectively, representative echocardiograms in LPS plus anti-MIF treated mice at 4 and 48 hours. A significant protection in cardiac function (FS %) is observed in LPS challenged mice when anti-MIF anti-bodies are given pre-treatment (E). Data represents the average of 9 cardiac cycles from 3 mice monitored at multiple time points. *p ⁇ 0.05.
- FIG. 19 Serum cytokine determination in wild type and TNFR ⁇ / ⁇ mice after LPS challenge.
- FIG. 20 Serum cytokine determination in wild type after LPS challenge with or without anti-MIF pre-treatment (90 minutes). Shown are the modulated cytokines: (A) IFN- ⁇ and (B) IL-10 as assayed on the Luminex platform. Data are expressed as the mean+/ ⁇ standard error of serum cytokine concentrations from 3 independent experimental mice at each time point.
- FIG. 21 Compares cardiac function (fractional shortening) in post LAD ligation with LAD only and anti-MIF+LAD.
- FIG. 22 Shows the effect of anti-MIF therapy pre-LAD with LAD only and anti-MIF+LAD.
- FIG. 23 Presents cardiac function data 48 hours post-LAD for several treatment groups.
- FIG. 24 Shows the serum troponin concentration 48 hrs post-LAD with pre- and delayed anti-MIF treatment.
- FIG. 25 Shows the serum troponin I and MIF concentrations through two weeks post ligation.
- FIG. 26 Shows the organ CD74 constitutive expression and heart CD74 post-LPS challenge expression.
- FIG. 28 Shows post LPS challenge serum sCD74 release.
- FIG. 29 Shows the CD74 series gel. See also Table 5.
- FIG. 30 Shows the coronary flow rate v. LVP, +dP/dt max, and ⁇ dP/dt max in CD74 KO Mouse.
- FIG. 31 Shows the hours post ligation v. fractional shortening for control, small molecule MIF inhibitor+DMSO/MI and DMSO/MI.
- One preferred embodiment of the invention relates to a method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor.
- MIF macrophage migration inhibitory factor
- One preferred embodiment of the invention relates to a method of treatment and/or prevention of cardiac dysfunction associated with burn injury.
- One preferred embodiment of the invention relates to the modulation of MIF as therapy for myocardial infarction.
- One preferred embodiment of the invention relates to the modulation of TNF- ⁇ in cardiac dysfunction.
- One preferred embodiment of the invention relates to a method of cardioprotection by inhibition of CD74.
- One preferred embodiment of the invention relates to the modulation of MIF with anti-MIF antibodies.
- One preferred embodiment of the invention relates to a bioassay for identifying agents that inhibit MIF activity.
- One preferred embodiment of the invention relates to the inhibition of MIF with anti-MIF antibody and concomitant restoration of (post-burning) cardiac function.
- One preferred embodiment of the invention relates to a method for the improvement of burn-injury-associated cardiac depression by the administration of anti-MIF antibodies.
- One preferred embodiment of the invention relates to the improvement of cardiac function following acute myocardial infarction by the administration of anti-MIF antibody.
- One preferred embodiment of the invention relates to the observation that MIF release from the heart, liver and spleen is dependent upon TNF- ⁇ receptor I/II signaling, and thus TNF- ⁇ may be a therapeutic target.
- One preferred embodiment of the invention relates to the neutralization of TNF- ⁇ with recombinant human TNFR:Fc.
- One preferred embodiment of the invention relates to the neutralization of MIF activity with one or more anti-MIF antibodies.
- One preferred embodiment of the invention relates to the discovery of TNF- ⁇ as an upstream mediator of MIF.
- CD74 an MIF receptor
- One preferred embodiment of the invention relates to the inhibition of MIF and improving cardioprotection by inhibiting the CD74 receptor with one or more anti-CD74 monoclonal neutralizing antibodies.
- One preferred object of the invention relates to a method for treating and/or preventing cardiac dysfunction, such as an irregularity or depression in myocardial activity.
- the method preferably includes administering an effective amount of a composition comprising a macrophage migration inhibitory factor (MIF) inhibitor.
- MIF macrophage migration inhibitory factor
- the inhibitor can be an antibody.
- the inhibitor can affect a particular MIF activity including an enzymatic activity, such as tautomerase activity or oxidoreductase activity.
- One preferred embodiment of the invention relates to an assay for identifying agents that inhibit MIF activity.
- the assay preferably involves both a myocyte, either in vitro or in vivo, and MIF in the presence and in the absence of an agent that may inhibit MIF activity.
- the assay analyzes myocyte activity, for example, using such tools as immunochemistry or echocardiography, based on the presence of MIF and a potential inhibitor.
- One preferred embodiment of the invention relates to a method of using this assay to identify an agent that inhibits MIF activity comprising placing a myocyte and MIF in the presence of an agent that may inhibit MIF activity, and determining the effect on myocyte activity.
- the myocyte may be in vitro or in vivo and the effect may be measured utilizing immunochemistry or echocardiography.
- MIF is an inducer of myocardial dysfunction, which is known to contribute significantly to the morbidity and mortality of sepsis in humans.
- sepsis associated cardiac dysfunction is characterized by biventricular dilatation, decreased systolic contractility, and diminished diastolic relaxation. While not wishing to be bound by theory, it is believed that its pathogenesis is multifactorial, with systemic and myocardial derived cytokines such as tumor necrosis factor-alpha (TNF- ⁇ ) involved in inducing its onset.
- TNF- ⁇ tumor necrosis factor-alpha
- One embodiment of the present application is directed to identifying whether MIF or other cardiac derived proteins mediate, by paracrine or autocrine mechanisms, myocardial dysfunction in sepsis and other cardiac diseases.
- Screening microarray analysis of cardiac gene expression in mice suggests that MIF is expressed in the heart, and is differentially regulated after lipopolysaccharide (LPS)-challenge.
- LPS lipopolysaccharide
- the following examples were constructed to verify whether MIF was expressed by cardiomyocytes in vivo, whether this expression was altered by endotoxin challenge and that MIF had a physiologically important effect on cardiac function.
- Several of the examples herein demonstrate cardiac MIF expression in vivo, and determine that MIF depresses cardiac function in a sublethal endotoxin challenge in vivo.
- MIF is constitutively expressed in the normal myocardium, and is released by cardiomyocytes following endotoxin challenge, with cardiac tissue levels reaching a nadir 12 hours after challenge.
- Evidence supporting a delayed release is seen in the present application by western blot and immunohistochemistry demonstrating significant release at 12 h from cardiac and spleen tissue, and supported indirectly by the delayed onset of cardiac protection beginning at eight hours, and continuing thereafter.
- Treatment of LPS challenged mice with anti-MIF monoclonal antibodies significantly improves in vivo cardiac function as evidenced by improvement in left ventricular shortening fraction.
- Microarray data on cardiac gene expression highlights that MIF is also expressed in cardiac tissue.
- MIF perfusion directly depresses cardiac function in vitro; and moreover, treatment with either of two independent monoclonal antibodies directed against MIF mitigates late myocardial depression.
- Another preferred embodiment of the invention relates to a method for treating and/or preventing burn injury associated conditions, including but not limited to cardiac dysfunction, such as an irregularity or depression in myocardial activity.
- the method preferably includes administering an effective amount of a composition comprising a macrophage migration inhibitory factor (MIF) inhibitor.
- MIF macrophage migration inhibitory factor
- the inhibitor can inhibit MIF activity and/or MIF production.
- the inhibitor is preferably an antibody or protein.
- the inhibitor can affect a particular MIF activity including an enzymatic activity, such as tautomerase activity or oxidoreductase activity.
- the inhibitor can inhibit or block MIF activity or MIF production in myocardial tissue.
- the inhibitor can also inhibit or prevent MIF release, such as inhibitors of the ABC transporter.
- Another preferred embodiment of the invention relates to an assay for identifying agents that inhibit MIF activity or production.
- the assay would involve a myocyte, either in vitro or in vivo, and possibly MIF in the presence and in the absence of an agent that inhibits MIF activity or MIF production.
- the assay would analyze myocyte activity, using such tools as immunochemistry or echocardiography, based on the presence of MIF and a potential inhibitor.
- One embodiment of the invention utilizes the role of MIF in burn associated cardiac dysfunction in methods of prevention/treatment and diagnostic assays.
- a murine burn injury model (40% TBSA)
- the present inventors identified that constitutive cardiac MIF significantly decreased (2.1 fold) 8 hours after burn injury as determined by western blot analysis. Serum MIF was maximal at 4 hours after burn injury (2.2 fold increase). These patterns are consistent with MIF release from pre-formed cytoplasmic stores of cardiac and systemic origin following burn injury.
- mice were pre-treated with anti-MIF neutralizing monoclonal antibodies.
- mice with burn injury alone demonstrated a depressed left ventricular fractional shortening percentage (FS %) of 38.6+/ ⁇ 1.8% (Sham FS % 56.0+/ ⁇ 2.6%).
- Anti-MIF treated mice demonstrated a delayed improved cardiac function after burn injury, with complete recovery of function by 24 hours. This demonstrates that the cytokine MIF mediates late burn injury associated cardiac dysfunction, and also demonstrates that MIF is a pharmacologic target for the treatment of burn injury associated cardiac dysfunction as well as other MIF mediated complications such as ARDS associated with burn injury.
- LPS LPS mediates the associated cardiac dysfunction through its interaction with the Tlr-4 (toll-like receptor 4) and its interaction with IRAK-1.
- the source of LPS is believed to be gut-derived due to several potential insults associated with burn injury. These include intestinal ischemia, bacterial translocation, and increased intestinal permeability. While not wishing to be bound by theory, it is thought that the production and release of inflammatory factors becomes systemic through gut associated lymphoid tissues.
- MIF mediated late cardiac depressant effects in vivo. It is also determined that recombinant MIF induces an immediate cardiac depression ex vivo by Langendorff assay in an LPS-independent manner.
- one embodiment of the present invention suitably has several advantages compared to observations made in a model of endotoxicosis. First, MIF is released at an earlier time point in the burn model (4 hours, FIG. 7 ) compared to the LPS challenge (8 hours) and this increase in systemic MIF concentrations is significantly higher (2.2 fold increase ( FIG. 9 ) vs. 1.5 fold).
- the degree of cardiac dysfunction was not as great in the burn injury model compared to the endotoxicosis model as measured by echocardiography.
- the present inventors desirably allows fractional shortening percent (an estimate of cardiac output) to be decreased 38% from baseline by four hours (56.2 FS % ⁇ 34.8 FS %/56.2 FS %) compared to 53.7% in the endotoxicosis model (67.2 FS % ⁇ 31.1 FS %/67.2 FS %) at four hours.
- MIF inhibition in accordance with one embodiment of the present invention results in complete cardiac protection by 24 hours ( FIG.
- TNF- ⁇ , IL-1 ⁇ , IL-6, and IL-10 are secreted by cardiomyocytes and TNF- ⁇ and IL-1 ⁇ are the primary mediators of the myocardial depression.
- the present invention contemplates the role these play in early cardiac dysfunction occurring before MIF mediated cardiac dysfunction. Since MIF is released locally from the heart just prior (8 hours) to the protective effects of anti-MIF treatment (12 hours) seen by echocardiography, the present invention contemplates that MIF plays a significant role in the cardiac dysfunction seen at later time points in this model (12-48 hours after burn injury).
- MIF cardiac malondialdehyde
- the cardiac release of pre-formed MIF may be initiated with increases in oxidative stress in the heart which signals the release of MIF ( FIGS. 7 and 8 ) and upregulates its transcription ( FIG. 10 ) in order to replenish the stores in cardiomyocytes.
- MIF cytokines
- MIF cytokines
- cytokines have tightly controlled expression that is upregulated after stimulation.
- MIF exists preformed in substantial amounts and its expression relies not only on de novo protein synthesis, but also from pre-existing stores which are controlled by secretory mechanisms involving ABC transporters.
- the MIF gene does not encode for an N-terminal signal sequence whose role is to translocate it to the endoplasmic reticulum.
- MIF is located predominantly in the cytosol in small vesicles and the nucleus which are pinched off and released to the outside of cells. Necrotic cell damage therefore leads to a release of the pre-stored MIF.
- MIF release may be directly released from necrotic cells of the skin, since MIF has been identified in the skin and localized to the basal layers of the epidermis.
- MIF is upregulated and plays a pivotal role in the pathophysiology of the disease.
- Total body UV B exposure in vivo has been shown to increase MIF production, suggesting its involvement in tissue injuries.
- Injured epidermis and cultured fibroblasts also increase the expression of MIF which contributes positively to the wound healing process.
- Systemic levels of MIF may increase more quickly and dramatically (2.2 fold by 4 hours in the burn injury model vs. 1.5 fold increase by 8 hours in the endotoxicosis model) in this burn injury model compared to the endotoxicosis model due to factors involving MIF released from burn injured skin.
- MIF has been shown has been hypothesized to play a role in ARDS and lung complications of sepsis.
- Anti-MIF therapy has been shown to decrease pulmonary neutrophil accumulation in acute lung injury associated with sepsis.
- MIF is expressed in alveolar capillary endothelium and infiltrating macrophages from ARDS patients.
- MIF expression has been shown to form an amplifying loop with TNF- ⁇ effectively linking severe inflammation to these two cytokines in ARDS. Since ARDS is an important and common complication of burn injury, the present invention contemplates anti-MIF therapies that are useful in other than cardiac protective indications, and seriously affect outcomes.
- MIF is unique among cytokines because it has multiple enzymatic activities including oxidoreductase and tautomerase activity. Inhibition of its tautomerase activity has been shown to counteract known MIF activities such as its glucocorticoid override activities. Pharmacological inhibitors of MIF tautomerase activity have been developed for diseases anti-MIF therapies have been effective such as sepsis, asthma, atopic dermatitis, and acute respiratory syndrome (ARDS).
- ARDS acute respiratory syndrome
- the cytokine MIF plays a significant role in the late cardiac dysfunction associated with burn injury.
- MIF itself is a direct cardiac depressant and has a delayed release from the heart.
- the delayed release of MIF and development of inhibitors that potentially inhibit the activity of MIF make MIF a potential target for diseases such as burn injury associated with morbidity and mortality related to its cardio-pulmonary effects.
- the present inventors have found that MIF release from the heart, liver, and spleen is dependent upon TNF receptor I/II signaling after LPS challenge. Additionally, the present inventors identify TNF receptor I/II signaling independent release into the serum of MIF. Without TNF receptor signaling, MIF levels appear slightly delayed (12-24 hours compared to 8 hours in wild type) and slightly increased (1.7-2.3 fold baseline compared to 1.5 increase in wild type mice). Moreover, the TNF receptor independent MIF release in TNF receptor I/II deficient mice (TNFR ⁇ / ⁇ ) is sufficient to mediate cardiac dysfunction by at least 24 hours after LPS challenge despite the lack of MIF release from tissues which has been previously identified in wild type mice.
- the cytokine MIF is constitutively expressed in numerous cell types including lymphocytes, macrophages, and the anterior pituitary. Many tissues also contain MIF including the heart, lung, liver, adrenal, spleen, kidney, skin, muscle, thymus, skin, and testes. The mechanism of secretion has recently been described in LPS stimulated monocytes. Inhibitors of classical protein secretion such as monensin or brefelding A do not inhibit the secretion of MIF, suggesting a non-classical protein export route. When inhibitors of ABCA1 (ATP binding cassette A1) transporters (glyburide and probenicide) were given, MIF secretion did not occur.
- ABCA1 ATP binding cassette A1
- MIF is located predominantly in the cytosol in small vesicles and the nucleus which are pinched off and released to the outside of cells.
- This non-classical, vesicle-mediated secretory pathway has been shown to be a mechanism of secretion of other important inflammatory mediators such as HMGB1, which has been shown to play a significant role in inflammatory diseases and specifically sepsis.
- HMGB1 important inflammatory mediators
- the dependence of MIF secretion on TNF- ⁇ signaling in several tissues is described for the first time.
- MIF has numerous biological activities including glucocorticoid antagonist properties, catalytic properties which are regulated through the coactivator JAB1/CSN5 and the cell surface protein CD74/Ii chain. Specific secretion of MIF results after inflammatory stimuli such as endotoxin (LPS) and tumor necrosis factor, as well as hormones such as ACTH, and angiotensin II. In addition to immune cells, endocrine cells and some epithelial cells secrete MIF. Secretion is due to an enhancement of MIF expression and de novo synthesis as well as an induction of the release from pre-existing stores; both of which have been previously demonstrated in the heart.
- Cardiac MIF has been reported to be released maximally at 12 hours after LPS challenge in wild type mice. Serum MIF levels in wild type mice after LPS challenge maximally release at 8 hours, corresponding to early protection of cardiac dysfunction.
- TNF- ⁇ signaling is inhibited after LPS challenge by either Enbrel®(D pretreatment or in TNFR ⁇ / ⁇ mice, the MIF levels peak later and slightly higher, indicating that TNF- ⁇ has some control over serum MIF levels, but does not inhibit MIF release.
- Enbrel® D pretreatment or in TNFR ⁇ / ⁇ mice
- TNF- ⁇ signaling has been studied by other investigators, specifically in relationship to the cytokine IL-18 in endotoxemia models.
- IL-18 levels in the heart are not significantly changed, while wild type mice demonstrated significant increases in IL-18 levels.
- IL-18 is neutralized, this study demonstrated that the LPS induced cardiac dysfunction is reduced and that IL-18 appears to have downstream effects on tissue TNF- ⁇ , IL-1 ⁇ , as well as ICAM-1/VCAM-1 levels. While this study focused on the myocardial production and release of IL-18 in the myocardium, non-cardiac sources of IL-18 were not investigated. This study is believed to be similar to that of the present inventors in the TNR- ⁇ dependence on tissue (cardiac) production/release of IL-18.
- mice deficient in IL-6 have augmented expression of IL-1 ⁇ and TNF- ⁇ after LPS challenge and the present inventors contemplate that cardiac IL-6 suppresses the expression of proinflammatory mediators including itself by a negative feedback mechanism.
- TNF- ⁇ signaling occurs through 2 receptors, TNF- ⁇ receptor 1 and 2. These two pathways have divergent signaling pathways. The interaction of TNF- ⁇ and receptor 1 activates several signal transduction pathways including NF-KB, which the TNF- ⁇ receptor does not.
- TNF- ⁇ receptor 1 activates several signal transduction pathways including NF-KB, which the TNF- ⁇ receptor does not.
- the present inventors challenged mice in which IkB overexpression in the heart resulted in nearly complete NF-kB inhibition. By western analysis, the present inventors demonstrated that no release occurred in the same manner as the TNFR ⁇ / ⁇ mice at all time points tested (data not shown, identical to FIG. 2A ). These mice have circulating TNF- ⁇ equivalent to wild types (since NF-kB inhibition is cardiac specific).
- mice express MIF in the serum similar to wild type mice. Accordingly, the present invention contemplates that the TNF receptor 1 may mediate the tissue release seen in wild type mice from the heart. Since the phenotype of this heart is completely protected after LPS challenge by echo during the first 48 hours (data not shown), the present inventors contemplate that circulating MIF requires upstream NF-kB mediated proteins to be signal (TNF- ⁇ , IL-1 ⁇ ) or that MIF mediates its effects by NF-kB itself.
- IFN- ⁇ was signficantlly suppressed.
- the expression of iNOS in cardiac myocytes have been shown to expressed when TNF- ⁇ and LPS are given with IFN- ⁇ , but NOT without IFN- ⁇ . Since iNOS itself plays a role in the regulatory pathways of LPS associated cardiac dysfunction, these cytokine pathways are complex and likely interact closely.
- MIF has been shown to be physiologically relevant in a model of live polymicrobial peritonitis (CLP).
- CLP live polymicrobial peritonitis
- MIF release is delayed and partially unaffected by blocking of important upstream mediators such as TNF- ⁇ , it may represent one good therapeutic target.
- the inhibition of MIF's tautomerase activity that mediate some of its biological functions may be intervened pharmacologically.
- recent non-classical secretory mechanisms of release also are potential target for therapy.
- HMGB1 significant targets in sepsis that are known
- undescribed targets still exist. Therefore, it is important to understand the effects of therapeutic invention of all of these mediators.
- MIF plays a significant role in LPS induced cardiac dysfunction which is believed to contribute to myocardial dysfunction during sepsis.
- TNF- ⁇ is thought to be an important sentinel cytokine in LPS induced cardiac dysfunction
- the effects of blocking TNF- ⁇ signaling pathways in vivo on MIF induced cardiac dysfunction have been investigated. Serum concentrations and the temporal distribution of MIF was slightly increased and delayed by the inhibition of TNF- ⁇ signaling (maximally increased 12-24 hours (1.7-2.3 fold baseline with TNF- ⁇ signaling inhibition vs. 8 hours in wild type (1.5 fold increase).
- Macrophage migration inhibitory factor is pluripotent cytokine with direct and significantly deleterious effects on heart function during sepsis (severe infections).
- MIF Macrophage migration inhibitory factor
- One embodiment of the present invention demonstrates in a mouse model that the inhibition of MIF activity can profoundly improve cardiac function following acute myocardial infarction (See FIGS. 21-25 ). This improvement is evident within hours, and lasts for the duration of the experiment (1 week). It is highly likely that modulation of MIF will decrease infarct size and other pathologic parameters. The degree of improvement in cardiac function is remarkable, and substantially in excess by at least 10 fold compared to other immune targets such as TNF-alpha.
- One embodiment of the present invention solves the problems of acute and chronic heart dysfunction following acute myocardial infarction.
- the present invention makes it possible to provide a unique class of therapies in that it modulates an immune mediator, i.e., MIF.
- MIF an immune mediator
- the present invention may directly decreases infarct size.
- modulation of MIF activity in accordance with the present invention would minimize the need for intra-aortic balloon pumps and other mechanical devices.
- Suitable monoclonal anti-MIF antibodies are obtained from Cytokine PharmaScience, Inc., King of Prussia, Pa.
- Another embodiment of the present invention relates to inhibition of CD74 to protect cardiodysfunction associated with severe disease such as sepsis, trauma, acute MI, and congestive heart failure. While CD74 has been described on circulating immune cells and antigen presenting cells (in association with MHC Class II), until the present invention, the presence of CD74 in the heart (as well as other organs) has not been reported. More importantly, the function role of CD74 on cardiac function in physiological or disease processes has not previously been demonstrated. While CD74 has been shown to mediate MIF activity in vitro, this has not been confirmed independently and is restricted to fibroblasts and leukocytes.
- Suitable anti-human CD74 antibodies are available, for example, from BD Biosciences (product catalog numbers 555538 (Clone M-B741; Format Purified; Isotype Mouse IgG 2a , ⁇ ; W.S. No V CD74.4; Reactivity Human) and 555612 (Clone LN2; Format Purified; Isotype Mouse IgG 1 , K; W.S. No V CD74.3; Reactivity Human).
- the present invention makes it possible to use anti-cytokine therapy (including anti-MIF) in cardiac diseases. Preliminary experiments using anti-TNF therapy in sepsis models did not work. Additionally, MIF, the putative cytokine blocked by CD74 inhibition, is a cytokine that occurs later, and in our model of acute MI is increased for several weeks after the insult, allowing for intervention during any of that time.
- cytokine therapy can enhance performance and potentially lower the morbidity associated with each.
- MI reperfusion
- MI and sepsis inotropes
- both acute and chronic cardiogenic impairment may be attenuated and improve survival/outcomes.
- MIF is secreted from cardiomyocytes following LPS challenge, and directly mediates a late onset (>6 hours) cardiac dysfunction.
- CD74 was determined to be the MIF receptor, exerting effects via ERK1/2 intracellular signaling pathways.
- the present inventors challenged: 1) wild type mice (C57BL/6) with LPS; 2) wild type mice pre-treated with anti-CD74 monocolonal neutralizing antibodies; and challenged with LPS, and 3) CD74 knock-out mice with LPS (4 mg/kg). Serial echocardiography was performed and fractional shortening (FS %) was determined.
- Another embodiment of the present invention relates to the inhibition of MIF activity by use of one or more soluble MIF receptor or MIF receptor antagonist.
- soluble MIF receptor or MIF receptor antagonist As an example, with anti-TNF ⁇ therapies, REMICADETM or INFLIXIMABTM (antibody TNF ⁇ ) and ENBRELTM or ETANERCEPTTM (soluble TNF-receptor) are suitable.
- This method includes administering one or more of the soluble MIF receptor and/or MIF receptor antagonist in an effective amount for treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, or a combination thereof to a subject in need thereof.
- Another embodiment of the present invention relates to the use of small molecule MIF inhibitors (sometimes called “MIF antagonists” or “isoxazoline compounds”) in treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, or a combination thereof.
- MIF antagonists sometimes called “MIF antagonists” or “isoxazoline compounds”
- the use of the superscript on a substituent is to identify a substituent name (e.g., “R 2 ” is used to indicate an R 2 -named substituent), while the use of a subscript is used to enumerate the number of times a substituent occurs at that molecular site (e.g., “R 2 “or “(R) 2 ” both are used to indicate two substituents simply named as “R”).
- a suitable small molecule MIF inhibitor for use in the methods herein has the following Formula I: wherein:
- the compound of Formula I is a p-hydroxyphenyl-isoxazoline-containing compound, wherein each of R, R 1-4 , X and Y is H or —CH 2 -A, and Z is OR. More preferably, the compound of Formula I is an ester of (R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazolineacetic, particularly the acid methyl ester thereof (sometimes identified as “ISO-1” or “CPSI” or “CPSI-26” herein) which is also known as p-hydroxyphenol-isoxazoline methyl ester.
- the compound is an ester of 2- ⁇ 3-(4-hydroxy-phenyl)-4,5-dihydro-isoxazol-5-yl ⁇ -3-phenyl-propinoic acid, particularly the methyl ester thereof (identified as “ISO-2”).
- the present invention also relates to the pharmaceutically acceptable acid addition salts of the compounds of general Formulas I, II, or III.
- the acids which are used to prepare the pharmaceutically acceptable acid addition salts of the aforementioned base compounds of this invention are those which form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, such as the chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, acid citrate, tartrate, bitartrate, succinate, maleate, fumarate, glutamate, L-lactate, L-tartrate, tosylate, mesylate, gluconate, saccharate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate (i.e., 1,1′-
- the invention also relates to base addition salts of the small molecule MIF inhibitors.
- the chemical bases that may be used as reagents to prepare pharmaceutically acceptable base salts of those compounds of general Formulas I, II, or III that are acidic in nature are those that form non-toxic base salts with such compounds.
- Such non-toxic base salts include, but are not limited to those derived from such pharmacologically acceptable cations such as alkali metal cations (e.g., potassium and sodium) and alkaline earth metal cations (e.g., calcium and magnesium), ammonium or water-soluble amine addition salts such as N-methylglucamine-(meglumine), and the lower alkanolammonium and other base salts of pharmaceutically acceptable organic amines.
- the compounds and prodrugs of the small molecule MIF inhibitors can exist in several tautomeric forms, and geometric isomers and mixtures thereof. All such tautomeric forms are included within the scope of the present invention. Tautomers exist as mixtures of tautomers in solution. In solid form, usually one tautomer predominates. Even though one tautomer may be described, the present invention includes all tautomers of the small molecule MIF inhibitors.
- the present invention also includes atropisomers of the small molecule MIF inhibitors.
- Atropisomers refer to compounds of the small molecule MIF inhibitors that can be separated into rotationally restricted isomers.
- the small molecule MIF inhibitors may contain olefin-like double bonds. When such bonds are present, the small molecule MIF inhibitors exist as cis and trans configurations and as mixtures thereof.
- a “suitable substituent” is intended to mean a chemically and pharmaceutically acceptable functional group i.e., a moiety that does not negate the inhibitory activity of the small molecule MIF inhibitors. Such suitable substituents may be routinely selected by those skilled in the art.
- substituents include, but are not limited to halo groups, perfluoroalkyl groups, perfluoroalkoxy groups, alkyl groups, alkenyl groups, alkynyl groups, hydroxy groups, oxo groups, mercapto groups, alkylthio groups, alkoxy groups, aryl or heteroaryl groups, aryloxy or heteroaryloxy groups, aralkyl or heteroaralkyl groups, aralkoxy or heteroaralkoxy groups, HO—(C ⁇ O)— groups, amino groups, alkyl- and dialkylamino groups, carbamoyl groups, alkylcarbonyl groups, alkoxycarbonyl groups, alkylaminocarbonyl groups dialkylamino carbonyl groups, arylcarbonyl groups, aryloxycarbonyl groups, alkylsulfonyl groups, arylsulfonyl groups and the like.
- Preferred small molecule MIF inhibitors may be found in U.S. provisional application 60/556,440, filed Mar. 26, 2004, U.S. provisional application 60/296,478, filed Jun. 8, 2001; and U.S. application Ser. No. 10/164,630, filed Jun. 10, 2002, the entire contents of each of which is hereby incorporated by reference for all purposes.
- an effective amount of one or more small molecule MIF inhibitors and/or salts thereof is administered as active ingredient to a subject in need thereof.
- Combinations of small molecule MIF inhibitors are also possible.
- the small molecule MIF inhibitor compounds can also be administered in form of their pharmaceutically active salts and/or prodrugs as appropriate. Combinations of salts and/or prodrugs are possible, as are combinations of salt-forms and non-salt-forms of the small molecule MIF inhibitor.
- Another embodiment of the present invention relates to pharmaceutical compositions suitable in treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, or a combination thereof, which includes one or more small molecule MIF inhibitors and/or salts thereof as active ingredient and at least one pharmaceutically acceptable carrier, excipient, adjuvent and/or diluent.
- Another embodiment of the present invention relates to the administration, to a subject in need thereof, of an effective amount of a composition which includes at least one small molecule MIF inhibitor, salt thereof, and/or prodrug thereof and at least one anti-MIF antibody in treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, or a combination thereof.
- a composition which includes at least one small molecule MIF inhibitor, salt thereof, and/or prodrug thereof and at least one anti-MIF antibody in treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardi
- Another embodiment of the present invention relates to pharmaceutical compositions suitable in treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, or a combination thereof, which includes an effective amount of a combination of at least one small molecule MIF inhibitor, salt thereof, and/or prodrug thereof and at least one anti-MIF antibody, and at least one pharmaceutically acceptable carrier, excipient, adjuvent and/or diluent.
- Another embodiment of the present invention relates to the administration, to a subject in need thereof, of an effective amount of a composition which includes a combination of at least one small molecule MIF inhibitor, salt thereof, and/or prodrug thereof, at least one anti-TNFR antibody and at least one anti-MIF antibody in treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, or a combination thereof.
- a composition which includes a combination of at least one small molecule MIF inhibitor, salt thereof, and/or prodrug thereof, at least one anti-TNFR antibody and at least one anti-MIF antibody in treating and
- Another embodiment of the present invention relates to pharmaceutical compositions suitable in treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, or a combination thereof, which includes an effective amount of a combination of at least one small molecule MIF inhibitor, salt thereof, and/or prodrug thereof, at least one anti-TNFR antibody and at least one anti-MIF antibody, and at least one pharmaceutically acceptable carrier, excipient, adjuvent and/or diluent.
- Another embodiment of the present invention relates to the administration, to a subject in need thereof, of an effective amount of a composition which includes a combination of at least one small molecule MIF inhibitor, salt thereof, and/or prodrug thereof, at least one anti-CD-74 antibody and at least one anti-MIF antibody in treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, or a combination thereof.
- a composition which includes a combination of at least one small molecule MIF inhibitor, salt thereof, and/or prodrug thereof, at least one anti-CD-74 antibody and at least one anti-MIF antibody in
- Another embodiment of the present invention relates to pharmaceutical compositions suitable in treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, or a combination thereof, which includes an effective amount of a combination of at least one small molecule MIF inhibitor, salt thereof, and/or prodrug thereof, at least one anti-CD-74 antibody and at least one anti-MIF antibody, and at least one pharmaceutically acceptable carrier, excipient, adjuvent and/or diluent.
- Another embodiment of the present invention relates to the administration, to a subject in need thereof, of an effective amount of a composition which includes a combination of at least one small molecule MIF inhibitor, salt thereof, and/or prodrug thereof and at least one anti-CD-74 antibody in treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, or a combination thereof.
- Another embodiment of the present invention relates to pharmaceutical compositions suitable in treating and/or preventing cardiac dysfunction in a subject in need thereof, treating and/or preventing irregularity in myocardial activity in a subject in need thereof, treating and/or preventing depression in myocardial activity in a subject in need thereof, treating and/or preventing burn-injury associated cardiac dysfunction in a subject in need thereof, treating and/or preventing cardiac dysfunction following acute myocardial infarction in a subject in need thereof, treating and/or preventing cardiodepression in a subject in need thereof, or a combination thereof, which includes an effective amount of a combination of at least one small molecule MIF inhibitor, salt thereof, and/or prodrug thereof, at least one anti-CD-74 antibody, and at least one pharmaceutically acceptable carrier, excipient, adjuvent and/or diluent.
- the present inventors have found that inhibiting MIF and/or neutralizing MIF tautomerase activity provides an anti-cytokine/inflammation therapy against cardiac diseases. Inhibition of MIF improves cardiac function after myocardial infarction and makes it possible to help with the acute sequelae of myocardial infarctions, such as reducing cardiac dysfunction early in acute myocardial infarction and reducing the associated high morbidity and mortality.
- One advantage the present invention has over current technologies is that unlike therapeutic interventions in cardiac disease that focus on inotropes, the present invention, by treating and/or preventing the cause of the cardiac dysfunction, both acute and chronic cardiogenic impairment may be attenuated and improve survival/outcomes.
- One embodiment of the present invention relates to pharmaceutical compositions comprising at least one compound of the present invention as an active ingredient (and/or salt and/or prodrug thereof) and at least one pharmaceutically acceptable carrier, excipient, adjuvent and/or diluent.
- compositions of the present invention may be prepared in any conventional solid or liquid carrier or diluent and optionally any conventional pharmaceutically-made adjuvant at suitable dosage level in a known way.
- the preferred preparations are in administrable form which is suitable for oral application. These administrable forms, for example, include pills, tablets, film tablets, coated tablets, capsules, powders and deposits.
- the compounds of the present invention and/or pharmaceutical preparations containing said compounds may be administered by any appropriate means, including but not limited to injection (intravenous, intraperitoneal, intramuscular, subcutaneous) by absorption through epithelial or mucocutaneous linings (oral mucosa, rectal and vaginal epithelial linings, nasopharyngial mucosa, intestinal mucosa); orally, rectally, transdermally, topically, intradermally, intragastrally, intracutanly, intravaginally, intravasally, intranasally, intrabuccally, percutanly, sublingually, or any other means available within the pharmaceutical arts.
- compositions of the present invention containing at least one compound of the present invention or pharmaceutically acceptable salts thereof as an active ingredient, will typically be administered in admixture with suitable carrier materials suitably selected with respect to the intended form of administration, i.e. oral tablets, capsules (either solid-filled, semi-solid filled or liquid filled), powders for constitution, oral gels, elixirs, dispersible granules, syrups, suspensions, and the like, and consistent with conventional pharmaceutical practices.
- suitable carrier materials suitably selected with respect to the intended form of administration, i.e. oral tablets, capsules (either solid-filled, semi-solid filled or liquid filled), powders for constitution, oral gels, elixirs, dispersible granules, syrups, suspensions, and the like, and consistent with conventional pharmaceutical practices.
- compositions may be comprised of from about 5 to about 95 percent by weight of the active ingredient, which range includes all values and subranges therebetween, including 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 90.5, 91, 91.5, 92, 92.5, 93, 93.5, 94, 94.5, and 95% by weight.
- Suitable binders include starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums such as acacia, sodium alginate, carboxymethyl-cellulose, polyethylene glycol and waxes.
- lubricants there may be mentioned for use in these dosage forms, boric acid, sodium benzoate, sodium acetate, sodium chloride, and the like.
- Disintegrants include starch, methylcellulose, guar gum and the like. Sweetening and flavoring agents and preservatives may also be included where appropriate.
- the compounds or compositions of the present invention may be formulated in sustained release form to provide the rate controlled release of any one or more of the components or active ingredients to optimize the therapeutic effects, i.e. antihistaminic activity and the like.
- Suitable dosage forms for sustained release include layered tablets containing layers of varying disintegration rates or controlled release polymeric matrices impregnated with the active components and shaped in tablet form or capsules containing such impregnated or encapsulated porous polymeric matrices.
- Liquid form preparations include solutions, suspensions and emulsions. As an example may be mentioned water, ethanolic, water-ethanol or water-propylene glycol solutions for parenteral injections or addition of sweeteners and opacifiers for oral solutions, suspensions and emulsions. Liquid form preparations may also include solutions for intranasal administration.
- Aerosol preparations suitable for inhalation may include solutions and solids in powder form, which may be in combination with a pharmaceutically acceptable carrier such as inert compressed gas, e.g. nitrogen.
- a pharmaceutically acceptable carrier such as inert compressed gas, e.g. nitrogen.
- the compounds of the present invention may also be deliverable transdermally.
- the transdermal compositions may take the form of creams, lotions, aerosols and/or emulsions and can be included in a transdermal patch of the matrix or reservoir type as are conventional in the art for this purpose.
- capsule refers to a special container or enclosure made of methyl cellulose, polyvinyl alcohols, or denatured gelatins or starch for holding or containing compositions comprising the active ingredients.
- Hard shell capsules are typically made of blends of relatively high gel strength bone and pork skin gelatins.
- the capsule itself may contain small amounts of dyes, opaquing agents, plasticizers and preservatives.
- Powders for constitution refers to powder blends containing the active ingredients and suitable diluents which can be suspended in water or juices.
- Suitable diluents are substances that usually make up the major portion of the composition or dosage form.
- Suitable diluents include sugars such as lactose, sucrose, mannitol and sorbitol, starches derived from wheat, corn rice and potato, and celluloses such as microcrystalline cellulose.
- the amount of diluent in the composition can range from about 5 to about 95% by weight of the total composition, preferably from about 25 to about 75%, more preferably from about 30 to about 60% by weight.
- disintegrants refers to materials added to the composition to help it break apart (disintegrate) and release the medicaments.
- Suitable disintegrants include starches, “cold water soluble” modified starches such as sodium carboxymethyl starch, natural and synthetic gums such as locust bean, karaya, guar, tragacanth and agar, cellulose derivatives such as methylcellulose and sodium carboxymethylcellulose, microcrystalline celluloses and cross-linked microcrystalline celluloses such as sodium croscarmellose, alginates such as alginic acid and sodium alginate, clays such as bentonites, and effervescent mixtures.
- the amount of disintegrant in the composition can range from about 2 to about 20% by weight of the composition, more preferably from about 5 to about 10% by weight.
- Glidents are materials that prevent caking and improve the flow characteristics of granulations, so that flow is smooth and uniform.
- Suitable glidents include silicon dioxide and talc.
- the amount of glident in the composition can range from about 0.1% to about 5% by weight of the total composition, preferably from about 0.5 to about 2% by weight.
- Coloring agents are excipients that provide coloration to the composition or the dosage form. Such excipients can include food grade dyes and food grade dyes adsorbed onto a suitable adsorbent such as clay or aluminum oxide. The amount of the coloring agent can vary from about 0.1 to about 5% by weight of the composition, preferably from about 0.1 to about 1%.
- a suitable composition comprising at least one compound of the invention may be a solution of the compound in a suitable liquid pharmaceutical carrier or any other formulation such as tablets, pills, film tablets, coated tablets, dragees, capsules, powders and deposits, gels, syrups, slurries, suspensions, emulsions, and the like.
- treating and/or preventing refers to reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which the term applies, or one or more symptoms of the disorder or condition.
- treatment refers to the act of treating and/or preventing as the term is defined above.
- the treated or administered subject is a human subject and more preferably a human subject in need of treatment.
- ⁇ ективное amount or “therapeutically effective amount” means an amount sufficient to cause any observable or measurable difference and preferably improvement in a subject's condition or indication, and preferably that condition or indication sought to be treated.
- an “acute” condition e.g. acute myocardial infarction
- a “chronic” condition e.g., chronic congestive heart failure
- antibody suitably includes antibody-derived fragment(s), Fab, Fab fragment(s), Fab 2 , CDR-derived regions, antibody-derived peptides, and/or single chain antibodies) as is known to those of ordinary skill in this art. Fab is preferred.
- cardiac dysfunction may suitably include one or more indications selected from the group including cardiac dysfunction, irregularity in myocardial activity, depression in myocardial activity, burn-injury associated cardiac dysfunction, cardiac dysfunction following acute myocardial infarction, cardiodepression, and a combination thereof. These terms are understood by a physician of ordinary skill in this art.
- a pharmaceutical composition for the treatment and prevention of cardiac dysfunction in a subject including:
- Goat anti-hMIF IgG and rhMIF were reconstituted in PBS and 0.1% BSA in PBS respectively, aliquoted, and stored at ⁇ 20° C. until use.
- Rabbit anti-goat IgG-HRP BioRad Corp., Hercules, Calif. stored at 4° C. until use.
- C57BL/6J and C3H/HeJ mice were obtained at 6-10 weeks of age (Jackson Labs, Bar Harbor, Me.).
- Adult Sprague-Dawley rats (Harlan Laboratories, Houston, Tex.) weighing 325-360 g were used in this study.
- Commercial chow and tap water were made available ad libitum. All animal protocols were reviewed and approved by the University of Texas Southwestern Medical Center Institutional Animal Care Advisory Committee and were in compliance with the rules governing animal use as published by the NIH.
- C57BL/6J mice were injected i.p. with 4 mg/kg E. coli 0111:B4 LPS (Sigma-Aldrich Corp., St.
- mice were used as controls.
- Two anti-MIF antibodies (III.D.9 and XIV.15.5, Rockland Immunochemicals, Inc., Gilbertsville, Pa.) and their isotype control (HB-49, Rockland Immunochemicals, Inc., Gilbertsville, Pa.) were injected (100 ⁇ g in 200 ⁇ l PBS) i.p. 90 m before the LPS challenge in the echocardiogram studies.
- Whole hearts were removed and snap frozen in liquid nitrogen and stored at ⁇ 80° C. or fixed in 10% neutral-buffered formalin for 24 h and placed in 70% ethanol for immunohistochemistry.
- the gel was transferred to PVDF membranes (NEN, Boston, Mass.) using a semi-dry transfer apparatus (Bio-Rad, Hercules, Calif.) at 15 V for 15 m.
- Membranes were blocked with TBS/0.1% Tween-20 (TBS-T) with 0.5% nonfat dry milk for 30 in and incubated with goat anti-hMIF IgG (1:750) in TBS/0.1% Tween-20/5% nonfat milk overnight at 4° C.
- TBS-T TBS/0.1% Tween-20
- goat anti-hMIF IgG 1:750
- TBS/0.1% Tween-20/5% nonfat milk overnight at 4° C.
- the membranes were washed 3 times for 10 m in TBS-T, incubated with rabbit anti-goat IgG-HRP (1:1000) for 1 h at RT, and washed 4 times for 10 m with TBS-T.
- the membranes were exposed to 5 ml of a mixture of luminol plus hydrogen peroxide under alkaline conditions (SuperSignal West Pico, Pierce, Rockford, Ill.) for 5 min and the resulting chemiluminescent reaction was detected by Kodak X-OMAT AR Film (Eastman Kodak Co., Rochester, N.Y.).
- RNA Extraction was extracted with Trizol (Invitrogen, Carlsbad, Calif.) from hearts thawed on ice according to the manufacturer's protocol and quantified by spectrophotometry.
- a MIF specific Northern probe was prepared by isolating DNA (DNeasy Tissue Kit, Qiagen, Valencia, Calif.) from the MIF plasmid (Research Genetics, Huntsville, Ala.) and subsequently cutting it with ECOR1 and NOT I restriction enzymes (Fisher Scientific, Pittsburgh, Pa.).
- the resultant DNA was resolved on a 1.2% agarose gel, purified (GenElute Agarose Spin Columns, Supelco, Bellefonte, Pa.), labeled with 5 ⁇ l 32 P-dCTP (3000 Ci/mmol)(PerkinElmer, Boston, Mass.) using Ready-To-Go Labeling Beads (Amersham Pharmacia, Piscatany, N.J.), and purified in ProbeQuant Microcolumns (Amersham Pharmacia, Piscatany, N.J.) according to manufacturer's protocols.
- the membranes were washed twice for 30 m in 2 ⁇ SSC/0.1% SDS at 46° C., and washed twice for 30 m in 0.2 ⁇ SSC/0.1% SDS at 46° C., and detected by Kodak X-OMAT AR Film (Eastman Kodak Co., Rochester, N.Y.). The same membranes were then probed with radiolabeled ⁇ -actin to ensure equal loading of protein.
- Tissue was fixed in neutral buffered formalin and processed to paraffin and subsequently immunostained at RT on a BioTek Solutions TechmateTM 1000 automated immunostainer (Ventana Medical Systems, Arlington, Ariz.) using the Ultra-streptavidin biotin system with horseradish peroxidase and diaminobenzidine (DAB) chromogen (Signet Laboratories, Dedham, M A).
- Optimum primary antibody dilutions were predetermined using known positive control tissues (rat post-LPS challenge). Paraffin sections were cut at 3 ⁇ m on a rotary microtome, mounted on positively charged glass slides (POP100 capillary gap slides, Ventana Medical Systems, Arlington, Ariz.) and air-dried overnight.
- Sections were then deparaffinized in xylene and ethanol, quenched with fresh 3% hydrogen peroxide for 10 m to inhibit endogenous tissue peroxidase activity, and rinsed with deionized water. Sections were incubated in unlabeled blocking serum for 15 m to block nonspecific binding of the secondary antibody and then incubated for 25 m with either rabbit anti-MIF (1:400, Torrey Pines BioLabs, Inc., Houston, Tex.) diluted in 1% citrate buffer (BioPath, Oklahoma City, Okla.), or with buffer alone as a negative reagent control.
- the aorta was cannulated with PE50 tubing, the heart perfused in a retrograde manner through the aortic root with prefiltered, oxygenated Krebs-Hanseleit Buffer at a constant flow rate of 1.5 ml/m (T 37° C.) and a recirculating volume of 100 ml.
- the heart was placed in a water-jacketed chamber to maintain constant temperature and humidity.
- PE60 intratnedic polyethylene tubing was connected to a Statham pressure transducer inserted into the left ventricle (LV) to measure LV pressure. Temperature was monitored using a 27 G thertnistor needle inserted into the LV muscle.
- LV pressure and its first derivative (dP/dt), heart rate, and coronary perfusion were measured simultaneously with a multichannel Grass 7D polygraph (Grass Instruments, Quincy, M A). Cardiac function for all hearts was determined by plotting peak systolic LV pressure and ⁇ dP/dt,Ia, values against changes in coronary flow rate. Hearts were perfused with or without 20 ng/ml RMIF added to the perfusate.
- Echocardiograms to assess systolic function were performed using M-mode measurements. Mice were anesthestized with 5% isofluorane with 2.5 L/m O 2 for 20 seconds (until unconscience) followed by 2% isofluorane and O 2 for an average of 12-15 m. Hair was removed from the thorax and upper abdomen using Nair® hair remover after sitting for 3 m using gauze. Echocardiography measurements were obtained on anesthetized mice approximately 5-8 m after induction to allow any transient cardiac depression to pass. These transient, minimal changes in cardiac function detected by echocardiography have been reported using inhaled isofluorane, although FS (%) has been reported to be stable.
- Cardiac echocardiography was performed using a Hewlett-Packard Sonos 5500 (Agilent Technologies; Edmonton, Alberta, Canada) with a frame rate of 300-500 frames/s in a random and blinded manner.
- a 12 MHz linear transducer was placed on the left hemithorax interfaced with a layer of US transmission gel (Aquasonic 100, Parker Laboratories; Fairfield, N.J.).
- the two dimensional parastemal short-axis imaging plane guided LV M-mode tracings close to the papillary muscle level. Depth was set at a minimum of 2 cm with a sweep speed of 150 m/s. Tracings were printed on a Sony color printer (UP-5200, Sony).
- End diastole was defined as the maximal LV diastolic dimension
- end systole was defined as the peak of posterior wall motion.
- Systolic function was calculated from LV dimensions as fractional shortening (FS) as follows: FS (%): LVED ⁇ LVES/LVED ⁇ 100, as shown in FIG. 5A .
- Northern and Western data are expressed as mean ⁇ standard error and statistically analyzed using a One Way Analysis of Variance. Determination of significance between experimental and control groups was performed using the Tukey method (p ⁇ 0.05). Cardiac function determined by the Langendorff preparation is expressed as mean ⁇ standard error and separate analyses were performed for each of LVP, +dP/dt max , and ⁇ dP/dt max , as a function of treatment group and coronary flow rate using a Repeated Measures Analysis of Variance. A multiple comparison procedure employing the Bonferroni method was used to determine significant differences between groups (p ⁇ 0.05).
- Cardiac function determined by echocardiogram is expressed by fractional shortening % (LVED ⁇ LVES/LVEP ⁇ 100.) ⁇ standard deviation and analyzed using a One Way Repeated Measures Analysis of Variance. Additional comparisons were performed using the Tukey Test to determine significant differences between specific groups (p ⁇ 0.05). All statistical analyses were performed using SigmaStat 2.03 (SPSS Inc., Chicago, Ill.) and Microsoft Excel (Microsoft Corp., Seattle, Wash.).
- MIF protein is constituitively expressed by cardiac myocytes in vivo and is released in response to LPS Challenge.
- Both immunochemistry and Western analysis performed on cardiac tissue documented the presence of MIF in cardiac cells, including ventricular and atrial myocytes, under baseline control conditions ( FIGS. 1 and 2 ).
- both immunochemistry and immunoblot analysis document a significant decrease in cardiac tissue MIF following endotoxin. This decrease was most profound (75% decrease) at 12 h, but returned to near baseline control levels by 24 h.
- This expression pattern in the heart is similar to that witnessed in the liver and spleen ( FIG. 2 ), and consistent with the hypothesis that MIF is released from preformed stores within tissue following LPS challenge.
- RNA obtained from the hearts of either control mice or from LPS challenged mice at given time points indicates that MIF mRNA is constitutively expressed in control mice, and that after LPS challenge, no significant change in MIF mRNA concentration is detectable in whole heart preparations ( FIG. 3 ).
- MIF induces systolic and diastolic cardiac dysfunction.
- rMIF recombinant MIF
- Responses to MIF were determined in hearts from both C57BL/6J mice, and C3H/HeJ mice.
- C3H/HeJ mice are resistant to endotoxin (41-43), therefore controlling for the possibility that any depression observed might be due to trace endotoxin in the perfusate.
- Table 2 illustrates the responses of both mouse strains to retrograde aortic perfusion at 1.5 ml/m with control perfusate or perfusate containing 20 ng/ml recombinant MIF.
- Perfusion with MIF led to a significant decrease in LVP, +dP/dt max , and ⁇ dP/dt max in both mouse strains.
- FIG. 4 illustrates the effect of MIF over a range of coronary flow rates. There is a step-wise increase in contractile performance in all hearts regardless of experimental group assignment. Comparison of the MIF exposed hearts with control hearts revealed a downward shift in the function curves, indicating significant systolic and diastolic depression in response to 20 ng/ml rMIF (p ⁇ 0.05).
- Anti-MIF antibodies improve LPS-induced cardiac depression in vivo.
- serial echocardiography M-mode was performed on LPS challenged mice which had been pre-treated (90 minutes prior) with either anti-MIF monoclonal antibodies, an isotype control antibody, or no treatment ( FIG. 5 ).
- FS fractional shortening %
- mice injected with either anti-MIF monoclonal antibody demonstrated statistically significant recovery of FS % compared to LPS challenged groups receiving either no treatment or isotype antibody control ( FIG.
- Antibodies and cytokines A polyclonal rabbit anti-rat MIF IgG (Torrey Pines BioLabs, Inc., Houston, Tex.) was used for western immunoblot and immunohistochemistry. This antibody has previously been shown to cross react with murine MIF and was prepared as previously described (23).
- a polyclonal goat anti-rabbit IgG-HRP BioRad Corp., Hercules, Calif. was used as a secondary antibody for western immunoblots and was stored at 4° C.
- MIF IgG1 antibodies Two monoclonal mouse anti-mouse (and human) MIF IgG1 antibodies (XIV.15.5 and III.D.9, gift of Cytokine PharmaSciences, Inc.) and a monoclonal mouse IgG1 isotype control antibody (HB-49, gift from Cytokine PharmaSciences, Inc.) were used in the echocardiographic studies. In vivo neutralization of MIF activity by both the XIV.15.5 and III.D.9 clones have been previously demonstrated.
- mice Male C57BL/6J mice ages 6-10 weeks (Jackson Labs, Bar Harbor, Me.) were maintained in a specific pathogen free environment. Commercial chow and tap water were made available ad libitum. All animal protocols were reviewed and approved by the University of Texas Southwestern Medical Center Institutional Animal Care Advisory Committee and were in compliance with the rules governing animal use as published by the NIH. Mice were subjected to a 40% TBSA burn injury. Briefly, mice were anesthetized with isoflourane (1-2%) with 2.5 L/minute oxygen to effect. Hair was then removed from their back and sides using a surgical prep blade and 70% ethanol. Brass probes heated to 100° C.
- mice received anesthesia and were shaven but not given the burn injury.
- Intraperitoneal injection of Lactated Ringer's with Buprenex (2 cc LR+0.2 cc Buprenex ( 0.05 mg/kg)) was given after the burn injury after the anesthesia was removed (with oxygen continued).
- Mice were then placed in individual cages under a heat lamp for approximately 1 hour and on a heating pad for the duration of the study and monitored closely. Mice were sacrificed at time points indicated in the figures by CO 2 asphyxiation followed by cervical dislocation.
- Monoclonal anti-MIF antibodies (III.D.9 and XIV.15.5) or an isotypic control (HB-49) were injected (100 ⁇ g in 200 ⁇ l PBS) intraperitoneally 90 minutes prior to burn injury in the echocardiogram studies.
- Whole hearts were removed, snap frozen in liquid nitrogen, and stored at ⁇ 80° C.
- hearts were fixed in 10% neutral-buffered formalin for 24 hours and were then placed in 70% ethanol until they were processed for immunohistochemistry.
- Whole blood was collected by retro-orbital bleeding and serum collected and stored.
- Prestained SDS-PAGE standards were run with each gel in order to determine the approximate M.W. of detected bands.
- the gel was transferred to a PVDF membrane (NEN, Boston, Mass.) using a mini transblot transfer apparatus (Bio-Rad, Hercules, Calif.) at 100 V for 70 minutes and cooled with ice packs.
- the membrane was re-wet with methanol, washed a minimum of 3 times with 100+ ml water, and blocked (5% nonfat dry milk (Bio-Rad)/TBS/0.1% Tween-20 (TBS-T) overnight at 4° C.
- the membrane was then incubated with the primary rabbit anti-MIF (1:1250 dilution) for 2 hours at room temperature in 5% milk/TBS-T and washed once for 15 minutes in TBS-T, followed by five washes (5 minutes each) in TBS-T. It was then incubated for 1 hour with a HRP conjugated goat anti-rabbit antibody in TBS-T (1:5000) at room temperature, washed twice for 15 minutes, followed by five additional washes (5 minutes each) in TBS-T.
- ECL reagent SuperSignal West Pico, Pierce, Rockford, Ill.
- chemiluminescent reaction was detected by Kodak X-OMAT AR Film (Eastman Kodak Co., Rochester, N.Y.).
- the quantification of the single band density with the approximate molecular weight of MIF (12.5 kD) was determined using Quantity One software (Bio-Rad, Hercules, Calif., Ver. 4.4.0, Build 36) following conversion of radiographic film to TIFF files (8 bit grayscale) using a Scanjet 3400c (Hewlett Packard, Palo Alto, Calif.) and reported in arbitrary units (A.U.)/mm 2 .
- Tissue was fixed in neutral buffered formalin, processed to paraffin, and subsequently immunostained at room temperature on a BioTek Solutions TechmateTM1000 automated immunostainer (Ventana Medical Systems, Arlington, Ariz.) using the Ultra-streptavidin biotin system with horseradish peroxidase and diaminobenzidine (DAB) chromogen (Signet Laboratories, Dedham, Mass.).
- Optimum primary antibody concentrations were predetermined using known positive control tissues (LPS challenged rat). Paraffin sections were cut at 3 ⁇ m on a rotary microtome, mounted on positively charged glass slides (POP100 capillary gap slides, Ventana Medical Systems, Arlington, Ariz.), and air-dried overnight.
- Sections were then deparaffinized in xylene and ethanol, quenched with fresh 3% hydrogen peroxide for 10 minutes to inhibit endogenous tissue peroxidase activity, and rinsed with de-ionized water. Sections were incubated in unlabeled blocking serum for 15 minutes to block nonspecific binding of the secondary antibody and then incubated for 25 minutes with either the polyclonal rabbit anti-rat MIF IgG (1:400, Torrey Pines BioLabs, Inc., Houston, Tex.) diluted in 1% citrate buffer (BioPath, Oklahoma City, Okla.) or with buffer alone as a negative reagent control. A negative reagent control was run for each time point and for each organ.
- TMB 3, 3′, 5, 5′-tetramethylbenzidene
- Plasma inflammatory cytokine (IL-1 ⁇ , IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IFN- ⁇ , TNF- ⁇ , and GM-CSF) concentrations were determined using the Mouse Cytokine Ten-Plex Antibody Bead Kit (Biosource International, Inc., Camarillo, Calif.) on a Luminex xMAP” system (Luminex Corp., Austin, Tex.) according to the manufacturer's instructions. The plate was loaded onto the Luminex XYPTM platform, the instrument set to remove 50 ⁇ l, and the total event set to equal the 100 per bead set.
- At least 100 events (most >200) for each cytokine were collected in each sample in order to determine statistically significant results.
- Data was collected using the LuminexTM Data Collector Software (Luminex Corp., Austin, Tex.). The concentrations of the lot specific reconstituted standards used in each run were entered into the software and the analyte concentrations for unknown samples were then extrapolated from the cytokine specific standard curve using MasterPlexTM QT software (Version 1.2.8.58, Mirai Bio, Inc., Alameda, Calif.). Final concentrations were multiplied by 2 in order to account for the initial dilution factor. No samples were detected that were higher than the standards curves for any cytokine.
- RNA isolation was prepared from an MIF containing plasmid (Image Clone I.D. 634910, Research Genetics, Huntsville, Ala.) isolated using Genelute HP Plasmid MidiPrep kit (Sigma, St. Louis, Mo.).
- the fragment was prepared by an EcoR1 and Not1 digestion (Fisher Scientific, Pittsburgh, Pa.) and gel purified and isolated on a 1.2% agarose gel using GenElute Agarose Spin Columns (Supelco, Bellefonte, Pa.).
- GenElute Agarose Spin Columns GenElute Agarose Spin Columns (Supelco, Bellefonte, Pa.).
- the ⁇ -actin probe DNA fragment was purchased from Ambion (Austin, Tex.).
- Both MIF and ⁇ -actin probes were labeled with 5 ⁇ l [ ⁇ - 32 P]dATP (3000 Ci/mmol, 10 mCi/ml) (PerkinElmer, Boston, Mass.) using Strip-EZTM DNA probe synthesis kit (Ambion, Austin, Tex.) and purified in ProbeQuant Microcolumns (Amersham Pharmacia, Piscatany, N.J.) according to manufacturers' protocols.
- RNA (10 ⁇ g) was resolved on 1.2% agarose gels at 100 volts for 1 hour and transferred to a Hybond-N+ membrane (Amersham Pharmacia, Buckingham, England) at 1.5 amps for 70 minutes on a transfer electrophoresis unit (TransPhor PowerLid, Hoefer Scientific Instruments, San Francisco). RNA was linked to the membrane for approximately 2 minutes using a GS Gene Linker (Bio-Rad, Hercules, Calif.). The membrane was prehybridized in a hybridization oven (Sorvall Life Science, Inc., Greensboro, N.C.) in Perfect-Hyb Plus (Sigma, St. Louis, Mo.) for 1 hour at 68° C.
- Sheared, denatured salmon or herring testis DNA (100 ⁇ g/ml) was then added for 1 hour, followed by the addition of approximately 0.1 ⁇ g probe labeled at >5 ⁇ 10 8 cpm/ ⁇ g.
- the blot was then hybridized for 12 hours at 68° C. in the hybridization oven followed by washing at 68° C. in 2 ⁇ SSC, 0.1% SDS.
- the membrane was washed for 1 hour, the buffer was exchanged, and then the membrane was washed for an additional hour at 68° C.
- the membrane was wrapped in Saran wrap, and mRNA was detected by Kodak X-OMAT AR Film after 24 hours (Eastman Kodak Co., Rochester, N.Y.).
- the aorta was cannulated with PE50 tubing, the heart perfused in a retrograde manner through the aortic root with pre-filtered, oxygenated Krebs-Henseleit Buffer at a constant flow rate of 1.5 ml/minute (T 37° C., 100 ml recirculating volume).
- the heart was placed in a water-jacketed chamber to maintain constant temperature and humidity.
- LV pressure and its first derivative (dP/dt), heart rate, and coronary perfusion were measured simultaneously with a multi-channel Grass 7D polygraph (Grass Instruments, Quincy, Mass.). Ventricular performance as a function of coronary perfusion was determined for all hearts by plotting peak systolic LV pressure and ⁇ dP/dt max values against incremental increases in coronary flow rate.
- Echocardiograms to assess systolic function were performed using M-mode measurements. Mice were anesthestized with 5% isofluorane with 2.5 L/m O 2 for 20 seconds (until unconscience) followed by 2% isofluorane and O 2 for an average of 12-15 minutes. Hair was removed from the thorax and upper abdomen using Nair® hair remover and gauze after sitting for 3 minutes. Echocardiographic measurements were obtained on anesthetized mice approximately 5-8 minutes after induction. Echocardiography was performed using a Hewlett-Packard Sonos 5500 (Agilent Technologies; Edmonton, Alberta, Canada) with a frame rate of 300-500 frames/second in a random and blinded manner.
- a 12 MHz linear transducer was placed on the left hemithorax interfaced with a layer of US transmission gel (Aquasonic 100, Parker Laboratories; Fairfield, N.J.).
- Northern and Western data are expressed as mean ⁇ standard error (SE) and statistically analyzed using a One Way-Analysis of Variance (ANOVA).
- SE mean ⁇ standard error
- ANOVA One Way-Analysis of Variance
- a multiple comparison procedure was employed using the Tukey method to determine statistical significance between groups.
- Cardiac function determined by the Langendorff preparation (including stabilization data) is expressed as the mean ⁇ SE and separate analyses were performed for each LVP, +dP/dt max , and ⁇ dP/dt max as a function of treatment group and coronary flow rate using a Repeated Measures-ANOVA.
- a multiple comparison procedure employing the Bonferroni method was used to determine significant differences between groups.
- Serum MIF levels are expressed as the mean ⁇ SE and were statistically analyzed using a One Way-ANOVA, with a multiple comparison procedure employing the Bonferroni method to determine significance between groups. Cardiac function determined by M-mode echocardiography is expressed as fractional shortening % ⁇ SE and analyzed using a One Way Repeated Measures-ANOVA. Additional comparisons were performed using the Tukey Test to determine significant differences between specific groups. Statistical significance for all analyses was defined as p ⁇ 0.05. All statistical analyses were performed using SigmaStat 2.03 (SPSS Inc., Chicago, Ill.) and Microsoft Excel (Microsoft Corp., Seattle, Wash.).
- MIF protein is constitutively expressed by cardiac myocytes in vivo and is released in response to burn injury.
- the cytokine macrophage migration inhibitory factor (MIF) is present in both ventricular and atrial myocytes at baseline as demonstrated by western and immunohistochemistry ( FIGS. 7 and 8 ).
- a significant decrease (2.1 fold) was identified at 8 hours with tissue concentrations of MIF returning to baseline levels by 12 hours ( FIG. 7 ).
- This expression pattern was paralleled in liver, spleen, and the lung after burn injury ( FIG. 8 ) and is consistent with the hypothesis that MIF is released in response mediators of burn injury.
- Systemic MIF and IL-6 levels are increased after burn injury.
- Maximum systemic release of MIF (2.2 fold increase) was identified in serum at 4 hours and returned to baseline levels by 8 hours ( FIG. 9 ).
- Maximum serum IL-6 levels were identified at 12 hours which returned to baseline levels by 48 hours.
- Serum IL-12 levels decreased after burn injury and were minimum at 24 hours and returned to baseline by 48 hours. No other cytokines tested (as listed in the materials and methods) were detected in the serum.
- MIF mRNA in the heart significantly increases in the heart by 8 hours after burn injury.
- the levels of MIF mRNA were detected by Northern analysis from total RNA isolated from hearts of either sham mice or mice at 4, 8, 12, 24, and 48 hours following burn injury ( FIG. 10 ).
- MIF mRNA is constitutively expressed in the heart, and significant increases in transcription initially occur at 8 hours, which are upregulated for the rest of the time course examined (48 hours) ( FIG. 10 ).
- Anti-MIF antibodies improve LPS-induced cardiac depression ex vivo.
- the responses of hearts to retrograde aortic perfusion at 1.5 ml/minute from mice undergoing the sham operation, burn injury, or burn injury with pre-treatment of anti-MIF antibodies were determined using a Langendorff analysis of heart function. Significant decreases in LVP, +dP/dt max , ⁇ dP/dT max , DR, dP40, TPP, RT90 and Time to Max ⁇ dP/dt were identified in mice 18 hours after undergoing burn injury (TABLE 3). Mice pre-treated with anti-MIF (Clone III.D.9) undergoing burn injury were completely protected by 18 hours (Table 3), while mice treated with the isotype control did not differ significantly from burn injury alone (data not shown).
- FIG. 11 illustrates the function of hearts over a range of coronary flow rates from sham mice, burn injury mice, and burn injury mice pre-treated with anti-MIF antibodies 18 hours after the burn injury or sham procedure.
- Increases in coronary flow resulted in incremental increases in contractile performance in all hearts (groups) tested.
- Mice undergoing burn injury demonstrated a downward shift in the LVP, +dP/dt max , and ⁇ dP/dT max function curves demonstrating significant systolic and diastolic dysfunction ( FIG. 11 ). This dysfunction, however, was not present when anti-MIF antibodies were given where no significant differences to sham mice were identified ( FIG. 11 ).
- Anti-MIF monoclonal antibody therapy improves burn injury associated cardiac depression in vivo.
- Serial echocardiography was (M-mode) was performed on mice receiving burn injury, and mice pre-treated 90 minutes prior before burn injury with either of two anti-MIF antibodies, an isotype control, or no treatment ( FIG. 12 ).
- FS % fractional shortening percentage of all burn injury treated mice were similarly depressed 21.4 FS (56.2 FS %-34.8 FS %), irrespective of anti-MIF treatment.
- mice injected with either of the two monoclonal anti-MIF antibodies demonstrated statistically significant recovery of FS % compared to burn injury mice receiving either no treatment of an isotype antibody control ( FIG. 12 ).
- mice receiving isotype control antibodies did not demonstrate significant differences from animals undergoing burn injury alone, indicating specificity of the anti-MIF antibody effects.
- Antibodies and cytokines A polyclonal rabbit anti-rat MIF IgG (Torrey Pines BioLabs, Inc., Houston, Tex.) which cross reacts with murine MIF was used for western immunoblot and immunohistochemistry. A polyclonal goat anti-rabbit IgG-HRP (BioRad Corp., Hercules, Calif.) was used as a secondary antibody for western immunoblots.
- mice were injected intraperitoneally with 4 mg/kg E. coli 0111:B4 LPS (Sigma-Aldrich Corp., St. Louis, Mo.) and sacrificed by CO 2 asphyxiation and subsequent cervical dislocation. Uninjected mice were used as controls.
- the two anti-MIF antibodies (III.D.9 and XIV.15.5, gift of Cytokine PharmaSciences, Inc., King of Prussia, Pa.) and their isotype control (HB-49, gift from Cytokine PharmaSciences, Inc.) were injected (100 ⁇ g in 200 ⁇ g PBS) intraperitoneally 90 minutes before the LPS challenge in the echocardiogram studies.
- Enbrel® (rhTNFR:Fc) was injected intraperitoneally (5 mg/kg or 300 ⁇ g in 0.5 ml PBS) 75 minutes prior to LPS challenge in wild type mice.
- the stop reagent was added to each well, gently mixed, and the ELISA was read on an ELISA plate reader (EL 312e Microplate Reader, Bio-Tek Instruments, Winooski, Vt.) at 450 nm (630 nm background) within 30 minutes of completion of the assay.
- ELISA plate reader EL 312e Microplate Reader, Bio-Tek Instruments, Winooski, Vt.
- Prestained SDS-PAGE standards (Kaleidoscope Broad range, Bio-Rad Laboratories, Inc., Hercules, Calif.) were run (10 ⁇ l/lane) with each gel in order to determine the approximate M.W. of detected bands.
- the gel was transferred to a PVDF membrane (NEN, Boston, Mass.) using the Mini Transblot® electrophoretic transfer cell (Bio-Rad, Hercules, Calif.) at 100 V for 70 minutes and cooled with ice packs.
- the membrane was re-wet with methanol, washed a minimum of 3 times with 100 ml water, and placed in block (5% nonfat dry milk (Bio-Rad)/TBS/0.1% Tween-20 (TBS-T) overnight at 4° C.
- the membrane was then incubated with the primary rabbit anti-MIF (1:1250 dilution) for 2 hours at room temperature in 5% milk/TBS-T.
- the membrane was washed 1 time for 15 minutes in TBS-T, followed by five washes (5 minutes each).
- the membrane was then incubated for 1 hour with a HRP conjugated goat anti-rabbit antibody (diluted 1:5000) in TBS-T at room temperature.
- the membrane was then washed twice for 15 minutes, followed by five additional washes (5 minutes each).
- the membrane was then developed for 5 minutes with 5 ml of ECL reagents (SuperSignal West Pico, Pierce, Rockford, Ill.), and the resulting chemiluminescent reaction was detected by Kodak X-OMAT AR Film (Eastman Kodak Co., Rochester, N.Y.).
- Quantification of the single band density with the approximate molecular weight of MIF (12.5 kD) was determined using Quantity One software (Bio-Rad, Hercules, Calif., Ver. 4.4.0, Build 36) following conversion of radiographic film to TIFF files (8 bit grayscale) using a Scanjet 3400c (Hewlett Packard, Palo Alto, Calif.) and reported in arbitrary units (A.U.)/mm 2 .
- Tissue was fixed in neutral buffered formalin and processed to paraffin and subsequently immunostained at room temperature on a BioTek Solutions TechmateTM1000 automated immunostainer (Ventana Medical Systems, Arlington, Ariz.) using the Ultra-streptavidin biotin system with horseradish peroxidase and diaminobenzidine (DAB) chromogen (Signet Laboratories, Dedham, Mass.). Paraffin sections were cut at 3 ⁇ m on a rotary microtome, mounted on positively charged glass slides (POP100 capillary gap slides, Ventana Medical Systems, Arlington, Ariz.) and air-dried overnight.
- DAB diaminobenzidine
- Sections were then deparaffinized in xylene and ethanol, quenched with fresh 3% hydrogen peroxide for 10 minutes to inhibit endogenous tissue peroxidase activity, and rinsed with deionized water. Sections were incubated in unlabeled blocking serum for 15 minutes to block nonspecific binding of the secondary antibody and then incubated for 25 minutes with either rabbit anti-MIF (1:400, Torrey Pines BioLabs, Inc., Houston, Tex.) diluted in 1% citrate buffer (BioPath, Oklahoma City, Okla.), or with buffer alone as a negative reagent control. Following washes in buffer, sections were incubated for 25 minutes with a biotinylated polyvalent secondary antibody solution (containing goat anti-rabbit antibodies).
- RNA isolation was prepared from an MIF containing plasmid (Image Clone I.D. 634910, Research Genetics, Huntsville, Ala.) isolated using Genelute HP Plasmid MidiPrep kit (Sigma, St. Louis, MO).
- the fragment was prepared by an EcoR1 and Not1 digestion (Fisher Scientific, Pittsburgh, Pa.) and gel purified and isolated on a 1.2% agarose gel using GenElute Agarose Spin Columns (Supelco, Bellefonte, Pa.).
- GenElute Agarose Spin Columns GenElute Agarose Spin Columns (Supelco, Bellefonte, Pa.).
- the ⁇ -actin probe DNA fragment was purchased from Ambion (Austin, Tex.).
- RNA Isolated total RNA (10 ⁇ g) was combined with formaldehyde loading dye (Ambion, Inc.) at a ratio of 1:3 sample:loading dye according to the manufacturer's protocols. Each gel had a 0.24-9.5 kB RNA ladder (Invitrogen Corp.) ran in parallel with samples (10 ⁇ g). Samples and RNA ladder were placed at 65° C.
- the membrane was prehybridized in a hybridization oven (Sorvall Life Science, Inc., Greensboro, N.C.) in Perfect-Hyb Plus (Sigma, St. Louis, Mo.) with sheared, denatured salmon sperm DNA (100 ⁇ g/ml) for 1 hour at 68° C.
- the probes were prepared by heating to 90° C. for 10 minutes (10 ⁇ l probe with 100 ⁇ l 10 mM EDTA), followed by the addition of approximately 0.1 ⁇ g probe labeled at >5 ⁇ 10 8 cpm/ ⁇ g to the hybridization buffer.
- the blot was then hybridized for 12 hours at 68° C. followed by washing at 68° C. in 2 ⁇ SSC, 0.1% SDS.
- the membrane was washed for 1 hour, the buffer was exchanged, and then the membrane was washed for an additional hour at 68° C.
- the membrane was wrapped in Saran wrap, and mRNA was detected by Kodak X-OMAT AR Film after 24 hours (Eastman Kodak Co., Rochester, N.Y.).
- the same membrane was then re-probed in a similar manner with radiolabeled ⁇ -actin (0.1 ⁇ g probe labeled at >5 ⁇ 10 8 cpM/ ⁇ g) (Ambion, Austin, Tex.). Densitometry was performed as described above for the western blots.
- the ⁇ -actin mRNA bands served as a control against which to normalize the MIF mRNA densitometry.
- the aorta was cannulated with PE50 tubing, the heart perfused in a retrograde manner through the aortic root with pre-filtered, oxygenated Krebs-Henseleit Buffer at a constant flow rate of 1.5 ml/minute (constant temperature of 37° C., 100 ml recirculating volume).
- the heart was placed in a water-jacketed chamber to maintain constant temperature and humidity.
- LV pressure and its first derivative (dP/dt), heart rate, and coronary perfusion were measured simultaneously with a multi-channel Grass 7D polygraph (Grass Instruments, Quincy, Mass.). Ventricular performance as a function of coronary perfusion was determined for all hearts by plotting peak systolic LV pressure and ⁇ dP/dt max values against incremental increases in coronary flow rate. Hearts were perfused with or without 20 ng/ml rMIF added to the perfusate.
- Echocardiograms to assess systolic function were performed using M-mode measurements. Mice were anesthestized with 5% isofluorane with 2.5 L/m O 2 for 20 seconds (until unconscience) followed by 2% isofluorane and O 2 for an average of 12-15 minutes. Hair was removed from the thorax and upper abdomen using Nair® hair remover and gauze after sitting for 3 minutes. Echocardiographic measurements were obtained on anesthetized mice approximately 5-8 minutes after induction.
- Echocardiography was performed using an Acuson SequoiaTM Model C256 (Siemens Medical Solutions, USA, Inc., Mountain View, Calif.) with a frame rate of 300-500 frames/second in a random and blinded manner.
- a 15 MHz linear transducer (15L8, Siemens Medical Solutions, USA, Inc.) was placed on the left hemithorax interfaced with a layer of ultrasound transmission gel (Aquasonic 100, Parker Laboratories; Fairfield, N.J.).
- the two dimensional parasternal short-axis imaging plane guided LV M-mode tracings close to the papillary muscle level. Depth was set at a minimum of 2 cm with a sweep speed of 200 m/second.
- Plasma inflammatory cytokine (IL-1 ⁇ , IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IFN- ⁇ , TNF- ⁇ , and GM-CSF) concentrations were determined using the Mouse Cytokine Ten-Plex Antibody Bead Kit (Biosource International, Inc., Camarillo, Calif.) on a Luminex xMAPTM system (Luminex Corp., Austin, Tex.) according to the manufacturer's instructions. The plate was loaded onto the Luminex XYPTM platform, the instrument set to remove 50 ⁇ l, and the total event set to equal the 100 per bead set (200 collected for most).
- LuminexTM Data Collector Software Luminex Corp., Austin, Tex.
- concentrations of the lot specific reconstituted standards used in each run were entered into the software and the analyte concentrations for unknown samples were then extrapolated from the cytokine specific standard curve using MasterPlexTM QT software (Version 1.2.8.58, Mirai Bio, Inc., Alameda, Calif.). Final concentrations were multiplied by 2 in order to account for the initial dilution factor. No samples were detected that were higher than the standards curves for any cytokine.
- Northern and Western data are expressed as mean ⁇ standard error (SE) and statistically analyzed using a One Way-Analysis of Variance (ANOVA).
- SE mean ⁇ standard error
- ANOVA One Way-Analysis of Variance
- a multiple comparison procedure was employed using the Tukey method to determine statistical significance between groups.
- Cardiac function determined by the Langendorff preparation (including stabilization data) is expressed as the mean ⁇ SE and separate analyses were performed for each LVP, +dP/dt max , and ⁇ dP/dt max as a function of treatment group and coronary flow rate using a Repeated Measures-ANOVA.
- a multiple comparison procedure employing the Bonferroni method was used to determine significant differences between groups.
- Serum MIF levels are expressed as the mean ⁇ SE and were statistically analyzed using a One Way-ANOVA, with a multiple comparison procedure employing the Bonferroni method to determine significance between groups. Cardiac function determined by M-mode echocardiography is expressed as fractional shortening % ⁇ SE and analyzed using a One Way Repeated Measures-ANOVA. Additional comparisons were performed using the Tukey Test to determine significant differences between specific groups. Statistical significance for all analyses was defined as p ⁇ 0.05. All statistical analyses were performed using SigmaStat 2.03 (SPSS Inc., Chicago, Ill.) and Microsoft Excel (Microsoft Corp., Seattle, Wash.).
- Serum MIF levels in WT mice, WT mice pre-treated with Enbrel®, and TNFR ⁇ / ⁇ mice Serum MIF levels in WT mice, WT mice pre-treated with Enbrel®, and TNFR ⁇ / ⁇ mice. Serum levels of MIF reach maximum ( ⁇ 1.5 fold baseline) at 8 hours in wild type mice after LPS challenge ( FIG. 13A ). When TNFR ⁇ / ⁇ mice are challenged with LPS, maximum serum MIF levels occur as 12 hours ( ⁇ 1.7 fold baseline)( FIG. 13B ). Maximum serum MIF levels ( ⁇ 2.3 fold baseline) were identified in wild type mice pre-treated (60 minutes) with Enbrel® and challenged with LPS at 24 hours ( FIG. 13C ).
- Cardiac MIF is not released from the heart, spleen, or liver in TNFR ⁇ / ⁇ mice after LPS challenge. Both western and immunohistochemistry analysis performed on cardiac, liver, and spleen demonstrated that previously documented release in wild type mice after LPS challenge did not occur at any time in TNFR ⁇ / ⁇ mice or wild type mice pre-treated with Enbrel®, both of which prevent TNF- ⁇ signaling ( FIGS. 14 and 15 ).
- MIF transcription is not modulated in TNFR ⁇ / ⁇ mice after LPS challenge. Detection of MIF mRNA from isolated total RNA from heart tissue from TNFR ⁇ / ⁇ mice challenge with LPS demonstrates that transcription of MIF is not upregulated after LPS challenge ( FIG. 16 ), which has been identified in wild type mice previously at 48 hours.
- MIF has direct cardiodepressant effects in TNFR ⁇ / ⁇ mice to the same extent as in WT mice.
- rMIF recombinant human MIF
- Table 4 demonstrates that the responses of the background strain of TNFR ⁇ / ⁇ mice, C57BL/6 mice, and the TNFR ⁇ / ⁇ mice to retrograde aortic perfusion at 1.5 ml/minute with control perfusate or perfusate containing 20 ng/ml rMIF.
- Perfusion with rMIF led to a significant decrease in LVP, +dP/dt max , ⁇ dP/dT max , and dp40 (mm Hg/sec) in both mouse strains while other parameters (time to max ⁇ dP/dt, CPP, CVR, and HR) were unaffected.
- FIG. 18 illustrates the effect of rMIF over a range of coronary flow rates.
- MIF neutralization by anti MIF antibodies results in complete protection at 24-48 hours after LPS challenge in TNFR ⁇ / ⁇ mice.
- serial echocardiography M-mode was performed on LPS challenged TNFR ⁇ / ⁇ mice pretreated (90 minutes prior) with either of two anti-MIF monoclonal antibodies, an isotype control antibody, or no treatment ( FIG. 17 ).
- FS fractional shortening percentage
- mice treated with either of the two monoclonal anti-MIF antibodies demonstrated statistically significant recovery of FS % compared to LPS challenged group receive or LPS or LPS and the isotype antibody ( FIG. 17 ).
- This enhanced recovery of function persisted at 48 hours where function was completely restored and LPS challenged mice receiving the isotype control were still profoundly depressed ( FIGS. 17D, 17E ).
- the FS % of untreated control TNFR ⁇ / ⁇ mice did not change significantly indicating that cardiac function was unaffected the testing regimen.
- Serum cytokine release in wild type mice and TNFR ⁇ / ⁇ mice Since other inflammatory cytokines have been shown to play a role in cardiac dysfunction in addition to TNF- ⁇ early after LPS challenge (i.e. IL-1 ⁇ , IL-6), we determined serum levels of an inflammatory panel in wild type and TNFR ⁇ / ⁇ mice ( FIG. 19 ). Not obvious in this figure are the release of TNF- ⁇ , IL-1 ⁇ in wild type mice because of the significant increases in these cytokines in TNFR ⁇ / ⁇ mice (31.2 (4934/158 pg/ml) fold and 94.7 (7099/75 pg/ml) fold increase over wild type at 4 hours after LPS challenge) as shown in FIGS. 19A and 19B .
- IL-12 was increased in the TNFR ⁇ / ⁇ mice compared to wild type mice (and 1.7 (5128/2937 pg/ml) fold) ( FIG. 19C ), while IFN- ⁇ levels were decreased 3.6 (210/58 pg/ml) fold ( FIG. 19D ).
- IL-10 and IL-6 increased similarly in wild type and TNFR ⁇ / ⁇ mice, although the delay of each of these cytokines was diminished in the TNFR ⁇ / ⁇ mice ( FIGS. 19E and 19F ).
- IL-6 levels were 8.5 fold in the TNFRKO mice at 4 hours compared to wild type mice (7099/835 pg/ml).
- Serum cytokine release after MIF neutralization in wild type mice is affected the modulation (increase or decrease) of serum IFN- ⁇ and IL-10 levels after LPS challenge in wild type mice ( FIG. 20 ). Specifically, the release of IFN- ⁇ peak at 8 hours after LPS challenge ( FIG. 19C ) was attenuated 3 fold (210/69 pg/ml)( FIG. 20A ). The delayed release of IL-10 in wild type mice which peaked at 48 hours was attenuated 2.9 fold (244/84 pg/ml)( FIG. 20B ) after LPS challenge in wild type mice.
- mice C57BL/6 mice from Charles River (12-15 weeks old) were maintained on commercial chow and tap water ad libitum. All animal protocols were reviewed and approved by the University of Texas Soiled IACAC in compliance with the rules governing animal use published by NIH.
- Coronary artery ligation Mice were anesthetized with 1-1.5% isoflurane after which coronary artery ligation was performed. Atropine (0.75 mg/kg given intramuscularly), lidocaine (1 mg/kg intramuscularly), and saline (1 ml intraperitoneally) were given pre-operatively. Ventilation was achieved using a custom mask fitted to the mouse snout and a small animal Ventilator (Harvard Apparatus, Inc., Holliston, Mass.). An incision ( ⁇ 5 mm) was made in the left thorax in the fourth intercostal space and pericardiotomy was performed to expose the left ventricle.
- the left coronary artery was occluded using 8-0 prolene approximately 2 mm under the left auricle. Subsequently, the chest was closed in layers and the negative pressure of the chest returned by syringe evacuation. Buprenorphine (0.10 mg/kg) was given once post-operatively for pain. Sham procedures were performed identically without the coronary ligation.
- Anti-MIF antibody A monoclonal anti-mouse MIF IgG1 antibodies (III.D.9, gift from Cytokine PharmaScience, Inc.) and a monoclonal IgG1 isotype control antibody (HB-49, gift from Cytokine PharmaScience, Inc.) were used in the echocardiographic studies. Previous studies have demonstrated in vivo neutralization of MIF activity.
- FIGS. 21-25 show the results obtained in this example.
- FIG. 21 Compares cardiac function (fractional shortening) in post LAD ligation with LAD only and anti-MIF+LAD.
- FIG. 22 Shows the effect of anti-MIF therapy pre-LAD with LAD only and anti-MIF+LAD.
- FIG. 21 Compares cardiac function (fractional shortening) in post LAD ligation with LAD only and anti-MIF+LAD.
- FIG. 22 Shows the effect of anti-MIF therapy pre-LAD with LAD only and anti-MIF+LAD.
- FIG. 23 Presents cardiac function data 48 hours post-LAD for several treatment groups.
- FIG. 24 Shows the serum troponin concentration 48 hrs post-LAD with pre- and delayed anti-MIF treatment.
- FIG. 25 Shows the serum troponin I and MIF concentrations through two weeks post ligation.
- MIF is secreted from cardiomyocytes following LPS challenge, and directly mediates a late onset (>6 hours) cardiac dysfunction.
- CD74 was recently determined to be the MIF receptor, exerting effects via ERK1/2 intracellular signaling pathways.
- To determine if CD74 mediates MIF-induced cardiac dysfunction in sepsis we challenged: 1) wild type mice (C57BL/6) with LPS; 2) wild type mice pre-treated with anti-CD74 monocolonal neutralizing antibodies; and challenged with LPS, and 3) CD74 knock-out mice with LPS (4 mg/kg). Serial echocardiography was performed and fractional shortening (FS %) was determined.
- Coronary artery ligation Mice were anesthetized with 1-1.5% isoflurane after which coronary artery ligation was performed. Atropine (0.075 mg/kg given intramuscularly), lidocaine (1 mg/kg intramuscularly), and saline (1 ml intraperitoneally) were given pre-operatively. Ventilation was achieved using a custom mask fitted to the mouse snout and a small animal ventilator (Harvard Apparatus, Inc., Holliston, Mass.). An incision ( ⁇ 5 mm) was made in the left thorax in the fourth intercostal space and pericardiotomy was performed to expose the left ventricle.
- the left coronary artery was occluded using 8-0 prolene approximately 2 mm under the left auricle. Subsequently, the chest was closed in layers and the negative pressure of the chest returned by syringe evacuation.
- An the acid methyl ester of (R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazolineacetic (“ISO-1”, “CPSI-26” or p-hydroxyphenol-isoxazoline methyl ester) was given at a dosage of 200 mg/kg in 25 microliters DMSO intraperitoneally daily for two weeks. Buprenorphine (0.10 mg/kg) was given once post-operatively for pain. Sham procedures were performed identically without the coronary ligation.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- Cardiology (AREA)
- Heart & Thoracic Surgery (AREA)
- Vascular Medicine (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Dermatology (AREA)
- Urology & Nephrology (AREA)
- Hospice & Palliative Care (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Nitrogen And Oxygen As The Only Ring Hetero Atoms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Priority Applications (22)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US10/927,494 US20050202010A1 (en) | 2003-08-29 | 2004-08-27 | Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor |
AU2004268017A AU2004268017B2 (en) | 2003-08-29 | 2004-08-30 | Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor |
PT47824271T PT1658037T (pt) | 2003-08-29 | 2004-08-30 | Método de tratamento e bioensaio envolvendo o fator de inibição da migração de macrófagos (mif) como fator depressivo do miocárdio de origem cardíaca |
PL04782427T PL1658037T3 (pl) | 2003-08-29 | 2004-08-30 | Sposób leczenia i test biologiczny z udziałem czynnika hamującego migrację makrofagów (mif) jako czynnika depresji mięśnia sercowego pochodzenia sercowego |
EP16170803.7A EP3078386A3 (en) | 2003-08-29 | 2004-08-30 | Method of treatment and bioassay involving macrophage migration inhibitory factor (mif) as cardiac-derived myocardial depressant factor |
MXPA06001828A MXPA06001828A (es) | 2003-08-29 | 2004-08-30 | Metodo para el tratamiento y bioensayo que involucra el factor de inhibicion de migracion de macrofagos ("mif"), como un factor de depresion del miocardio, derivado del corazon. |
CA2537928A CA2537928C (en) | 2003-08-29 | 2004-08-30 | Method of treatment and bioassay involving macrophage migration inhibitory factor (mif) as cardiac-derived myocardial depressant factor |
BRPI0413404-4A BRPI0413404A (pt) | 2003-08-29 | 2004-08-30 | composições farmacêuticas, métodos de tratamento ou prevenção de disfunções cardìacas, métodos de aumento da função cardìaca e método de identificação de inibidor de mif |
CN2004800247776A CN1972713B (zh) | 2003-08-29 | 2004-08-30 | 治疗或预防心脏功能障碍的药物组合物 |
ES04782427.1T ES2599032T3 (es) | 2003-08-29 | 2004-08-30 | Método de tratamiento y bioensayo que implica el factor de inhibición de la migración de macrófagos (MIF) como factor depresor del miocardio de origen cardiaco |
JP2006524893A JP4891769B2 (ja) | 2003-08-29 | 2004-08-30 | マクロファージ遊走阻止因子(mif)を心臓由来心筋抑制因子として含む、処置方法およびバイオアッセイ |
CN201110370579.7A CN102499984B (zh) | 2003-08-29 | 2004-08-30 | 涉及作为心脏衍生的心肌抑制因子的巨噬细胞迁移抑制因子(mif)的治疗和生物测定方法 |
DK04782427.1T DK1658037T3 (en) | 2003-08-29 | 2004-08-30 | Method and treatment of bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depression factor |
HUE04782427A HUE029888T2 (en) | 2003-08-29 | 2004-08-30 | Treatment Method and Biological Test Using Macrophage Migration Factor (MIF) as a Factor for Heart Myocardial Depression |
EP04782427.1A EP1658037B1 (en) | 2003-08-29 | 2004-08-30 | Method of treatment and bioassay involving macrophage migration inhibitory factor (mif) as cardiac-derived myocardial depressant factor |
PCT/US2004/027945 WO2005020919A2 (en) | 2003-08-29 | 2004-08-30 | Method of treatment and bioassay involving macrophage migration inhibitory factor (mif) as cardiac-derived myocardial depressant factor |
MX2011013436A MX340217B (es) | 2003-08-29 | 2006-02-16 | Metodo para el tratamietno y bioensayo que involucra el factor de inhibicion de migracion de macrofagos (mif), como un factor de depresion del miocardio, derivado del corazon. |
US11/932,909 US8747843B2 (en) | 2003-08-29 | 2007-10-31 | Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor |
JP2010039451A JP5905659B2 (ja) | 2003-08-29 | 2010-02-24 | マクロファージ遊走阻止因子(mif)を心臓由来心筋抑制因子として含む、処置方法およびバイオアッセイ |
JP2011133728A JP2011251966A (ja) | 2003-08-29 | 2011-06-15 | マクロファージ遊走阻止因子(mif)を心臓由来心筋抑制因子として含む、処置方法およびバイオアッセイ |
US14/281,870 US20150017179A1 (en) | 2003-08-29 | 2014-05-19 | Method of Treatment and Bioassay Involving Macrophage Migration Inhibitory Factor (MIF) as Cardiac-Derived Myocardial Depressant Factor |
CY20161100837T CY1117925T1 (el) | 2003-08-29 | 2016-08-25 | Μεθοδος αγωγης και βιοπροσδιορισμος που σχετιζεται με παραγοντα αναστολης της μεταναστευσης των μακροφαγων (mif) ως προερχομενο απο την καρδια μυοκαρδιακο κατασταλτικο παραγοντα |
Applications Claiming Priority (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US49865903P | 2003-08-29 | 2003-08-29 | |
US54705404P | 2004-02-25 | 2004-02-25 | |
US54705604P | 2004-02-25 | 2004-02-25 | |
US54705904P | 2004-02-25 | 2004-02-25 | |
US54705704P | 2004-02-25 | 2004-02-25 | |
US55644004P | 2004-03-26 | 2004-03-26 | |
US10/927,494 US20050202010A1 (en) | 2003-08-29 | 2004-08-27 | Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/932,909 Continuation US8747843B2 (en) | 2003-08-29 | 2007-10-31 | Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor |
Publications (1)
Publication Number | Publication Date |
---|---|
US20050202010A1 true US20050202010A1 (en) | 2005-09-15 |
Family
ID=34280266
Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US10/927,494 Abandoned US20050202010A1 (en) | 2003-08-29 | 2004-08-27 | Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor |
US11/932,909 Active 2027-03-21 US8747843B2 (en) | 2003-08-29 | 2007-10-31 | Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor |
US14/281,870 Abandoned US20150017179A1 (en) | 2003-08-29 | 2014-05-19 | Method of Treatment and Bioassay Involving Macrophage Migration Inhibitory Factor (MIF) as Cardiac-Derived Myocardial Depressant Factor |
Family Applications After (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/932,909 Active 2027-03-21 US8747843B2 (en) | 2003-08-29 | 2007-10-31 | Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor |
US14/281,870 Abandoned US20150017179A1 (en) | 2003-08-29 | 2014-05-19 | Method of Treatment and Bioassay Involving Macrophage Migration Inhibitory Factor (MIF) as Cardiac-Derived Myocardial Depressant Factor |
Country Status (15)
Country | Link |
---|---|
US (3) | US20050202010A1 (ja) |
EP (2) | EP1658037B1 (ja) |
JP (3) | JP4891769B2 (ja) |
CN (2) | CN102499984B (ja) |
AU (1) | AU2004268017B2 (ja) |
BR (1) | BRPI0413404A (ja) |
CA (1) | CA2537928C (ja) |
CY (1) | CY1117925T1 (ja) |
DK (1) | DK1658037T3 (ja) |
ES (1) | ES2599032T3 (ja) |
HU (1) | HUE029888T2 (ja) |
MX (2) | MXPA06001828A (ja) |
PL (1) | PL1658037T3 (ja) |
PT (1) | PT1658037T (ja) |
WO (1) | WO2005020919A2 (ja) |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20080241145A1 (en) * | 2004-12-08 | 2008-10-02 | Immunomedics, Inc. | Methods and compositions for immunotherapy and detection of inflammatory and immune-dysregulatory disease, infectious disease, pathologic angiogenesis and cancer |
WO2009117706A3 (en) * | 2008-03-20 | 2010-01-21 | Carolus Therapeutics, Inc. | Methods of treatment using anti-mif antibodies |
WO2010065491A2 (en) * | 2008-12-01 | 2010-06-10 | Carolus Therapeutics, Inc. | Methods of treating inflammatory disorders |
US20100183598A1 (en) * | 2008-11-12 | 2010-07-22 | Carolus Therapeutics, Inc. | Methods of treating cardiovascular disorders |
US20110070184A1 (en) * | 2008-03-24 | 2011-03-24 | Carolus Therpeutics, Inc. | Methods and compositions for treating atherosclerosis and related condidtions |
US9238689B2 (en) | 2011-07-15 | 2016-01-19 | Morpho Sys AG | Antibodies that are cross-reactive for macrophage migration inhibitory factor (MIF) and D-dopachrome tautomerase (D-DT) |
US20210270829A1 (en) * | 2018-08-30 | 2021-09-02 | Essen Instruments, Inc. D/B/A Essen Bioscience, Inc. | Methods For Determining Concentration Of Low And High Concentration Proteins In A Single Sample |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050202010A1 (en) | 2003-08-29 | 2005-09-15 | Giroir Brett P. | Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor |
CA2557166C (en) * | 2004-03-26 | 2015-06-30 | Cytokine Pharmasciences, Inc. | Compounds, compositions, processes of making, and methods of use related to inhibiting macrophage migration inhibitory factor |
WO2006116688A2 (en) * | 2005-04-26 | 2006-11-02 | Yale University | Mif agonists and antagonists and therapeutic uses thereof |
EP2742348A4 (en) * | 2011-08-12 | 2015-07-22 | Alfred Health | METHOD FOR THE DIAGNOSIS, FORECAST OR TREATMENT OF ACUTE CORONARY SYNDROME (ACS) BY MEASUREMENT OF THE PLASMA CONCENTRATION OF THE MACROPHAGE MIGRATION INHIBITION FACTOR (MIF) |
WO2013050453A1 (en) | 2011-10-07 | 2013-04-11 | Baxter Healthcare S.A. | Oxmif as a diagnostic marker |
WO2014086916A1 (en) * | 2012-12-07 | 2014-06-12 | Baxter International Inc. | Anti-mif antibody cell migration assay |
CN105087610A (zh) * | 2015-09-11 | 2015-11-25 | 中国科学院海洋研究所 | 文蛤巨噬细胞迁移抑制因子基因及其编码蛋白和应用 |
JP2018007029A (ja) * | 2016-07-01 | 2018-01-11 | 株式会社村田製作所 | バイアス回路 |
EP4334469A1 (en) * | 2021-05-06 | 2024-03-13 | Abbott Molecular Inc. | Compositions and methods for simple sample extraction |
CN114288387A (zh) * | 2022-02-17 | 2022-04-08 | 重庆医科大学 | Humanin衍生物HNG在制备治疗心衰药物中的应用 |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6080407A (en) | 1993-05-17 | 2000-06-27 | The Picower Institute For Medical Research | Diagnostic assays for MIF |
US6420188B1 (en) | 1996-02-16 | 2002-07-16 | The Picower Institute For Medical Research | Screening assay for the identification of inhibitors for macrophage migration inhibitory factor |
US6492428B1 (en) | 2000-07-26 | 2002-12-10 | The Picower Institute For Medical Research | Compounds having MIF antagonist activity |
CA2309115C (en) * | 1997-11-05 | 2010-10-19 | University Of Southern California | Use of cytokines and mitogens to inhibit pathological immune responses |
JP2003513065A (ja) | 1999-10-29 | 2003-04-08 | ザ・ピコワー・インスティチュート・フォー・メディカル・リサーチ | Mifアンタゴニスト活性を有する化合物 |
US6268151B1 (en) | 2000-01-20 | 2001-07-31 | Isis Pharmaceuticals, Inc. | Antisense modulation of macrophage migration inhibitory factor expression |
US20030235584A1 (en) * | 2000-02-28 | 2003-12-25 | Kloetzer William S. | Method for preparing anti-MIF antibodies |
AU2002255961B2 (en) * | 2001-03-29 | 2007-08-23 | Baxalta GmbH | Methods and compositions for using MHC class II invariant chain polypeptide as a receptor for macrophage migration inhibitory factor |
JP4410553B2 (ja) * | 2001-06-08 | 2010-02-03 | サイトカイン・ファーマサイエンシズ・インコーポレーテッド | Mifアンタゴニスト活性を有するイソオキサゾリン化合物 |
US20050202010A1 (en) | 2003-08-29 | 2005-09-15 | Giroir Brett P. | Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor |
-
2004
- 2004-08-27 US US10/927,494 patent/US20050202010A1/en not_active Abandoned
- 2004-08-30 ES ES04782427.1T patent/ES2599032T3/es not_active Expired - Lifetime
- 2004-08-30 PL PL04782427T patent/PL1658037T3/pl unknown
- 2004-08-30 EP EP04782427.1A patent/EP1658037B1/en not_active Expired - Lifetime
- 2004-08-30 CA CA2537928A patent/CA2537928C/en not_active Expired - Lifetime
- 2004-08-30 AU AU2004268017A patent/AU2004268017B2/en not_active Expired
- 2004-08-30 CN CN201110370579.7A patent/CN102499984B/zh not_active Expired - Lifetime
- 2004-08-30 HU HUE04782427A patent/HUE029888T2/en unknown
- 2004-08-30 DK DK04782427.1T patent/DK1658037T3/en active
- 2004-08-30 WO PCT/US2004/027945 patent/WO2005020919A2/en active Application Filing
- 2004-08-30 JP JP2006524893A patent/JP4891769B2/ja not_active Expired - Lifetime
- 2004-08-30 BR BRPI0413404-4A patent/BRPI0413404A/pt not_active Application Discontinuation
- 2004-08-30 MX MXPA06001828A patent/MXPA06001828A/es active IP Right Grant
- 2004-08-30 CN CN2004800247776A patent/CN1972713B/zh not_active Expired - Lifetime
- 2004-08-30 PT PT47824271T patent/PT1658037T/pt unknown
- 2004-08-30 EP EP16170803.7A patent/EP3078386A3/en not_active Withdrawn
-
2006
- 2006-02-16 MX MX2011013436A patent/MX340217B/es unknown
-
2007
- 2007-10-31 US US11/932,909 patent/US8747843B2/en active Active
-
2010
- 2010-02-24 JP JP2010039451A patent/JP5905659B2/ja not_active Expired - Lifetime
-
2011
- 2011-06-15 JP JP2011133728A patent/JP2011251966A/ja active Pending
-
2014
- 2014-05-19 US US14/281,870 patent/US20150017179A1/en not_active Abandoned
-
2016
- 2016-08-25 CY CY20161100837T patent/CY1117925T1/el unknown
Cited By (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20080241145A1 (en) * | 2004-12-08 | 2008-10-02 | Immunomedics, Inc. | Methods and compositions for immunotherapy and detection of inflammatory and immune-dysregulatory disease, infectious disease, pathologic angiogenesis and cancer |
US8420786B2 (en) * | 2004-12-08 | 2013-04-16 | Immunomedics, Inc. | Bispecific antibody targeting a complement factor or complement regulatory protein |
WO2009117706A3 (en) * | 2008-03-20 | 2010-01-21 | Carolus Therapeutics, Inc. | Methods of treatment using anti-mif antibodies |
EP2254597A2 (en) * | 2008-03-20 | 2010-12-01 | Carolus Therapeutics, Inc. | Methods of treatment using anti-mif antibodies |
US20110044988A1 (en) * | 2008-03-20 | 2011-02-24 | Carolus Therpeutics, Inc. | Methods of treatment using anti-mif antibodies |
EP2254597A4 (en) * | 2008-03-20 | 2012-04-18 | Carolus Therapeutics Inc | TREATMENT PROCEDURE WITH ANTI-MIF ANTIBODIES |
US20110070184A1 (en) * | 2008-03-24 | 2011-03-24 | Carolus Therpeutics, Inc. | Methods and compositions for treating atherosclerosis and related condidtions |
US20100183598A1 (en) * | 2008-11-12 | 2010-07-22 | Carolus Therapeutics, Inc. | Methods of treating cardiovascular disorders |
WO2010065491A2 (en) * | 2008-12-01 | 2010-06-10 | Carolus Therapeutics, Inc. | Methods of treating inflammatory disorders |
WO2010065491A3 (en) * | 2008-12-01 | 2010-09-30 | Carolus Therapeutics, Inc. | Methods of treating inflammatory disorders |
US9238689B2 (en) | 2011-07-15 | 2016-01-19 | Morpho Sys AG | Antibodies that are cross-reactive for macrophage migration inhibitory factor (MIF) and D-dopachrome tautomerase (D-DT) |
US20210270829A1 (en) * | 2018-08-30 | 2021-09-02 | Essen Instruments, Inc. D/B/A Essen Bioscience, Inc. | Methods For Determining Concentration Of Low And High Concentration Proteins In A Single Sample |
Also Published As
Publication number | Publication date |
---|---|
CA2537928A1 (en) | 2005-03-10 |
AU2004268017A2 (en) | 2005-03-10 |
PL1658037T3 (pl) | 2017-08-31 |
JP2007504158A (ja) | 2007-03-01 |
EP1658037A4 (en) | 2008-09-10 |
ES2599032T3 (es) | 2017-01-31 |
US20080260723A1 (en) | 2008-10-23 |
WO2005020919A3 (en) | 2006-11-30 |
CN102499984A (zh) | 2012-06-20 |
JP2011251966A (ja) | 2011-12-15 |
PT1658037T (pt) | 2016-08-30 |
WO2005020919A2 (en) | 2005-03-10 |
BRPI0413404A (pt) | 2006-10-17 |
CY1117925T1 (el) | 2017-05-17 |
US8747843B2 (en) | 2014-06-10 |
JP5905659B2 (ja) | 2016-04-20 |
MXPA06001828A (es) | 2007-05-23 |
JP2010159267A (ja) | 2010-07-22 |
CN1972713A (zh) | 2007-05-30 |
DK1658037T3 (en) | 2016-08-29 |
EP1658037A2 (en) | 2006-05-24 |
US20150017179A1 (en) | 2015-01-15 |
CA2537928C (en) | 2015-05-05 |
AU2004268017A1 (en) | 2005-03-10 |
JP4891769B2 (ja) | 2012-03-07 |
AU2004268017B2 (en) | 2011-03-17 |
EP3078386A3 (en) | 2017-01-11 |
CN102499984B (zh) | 2015-02-18 |
MX340217B (es) | 2016-06-30 |
EP1658037B1 (en) | 2016-05-25 |
EP3078386A2 (en) | 2016-10-12 |
CN1972713B (zh) | 2012-02-29 |
HUE029888T2 (en) | 2017-04-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US8747843B2 (en) | Method of treatment and bioassay involving macrophage migration inhibitory factor (MIF) as cardiac-derived myocardial depressant factor | |
JP6066474B2 (ja) | 中和プロラクチン受容体抗体およびそれらの治療的使用 | |
EP2170366B1 (en) | Use of tlr-2 antagonists for treatment of reperfusion injury and tissue damage | |
TWI834025B (zh) | 用於治療和/或預防冠狀病毒誘導的急性呼吸窘迫症候群的抑制masp-2的方法 | |
KR20180135928A (ko) | 간 질환을 치료하거나 예방하는 방법 | |
JP2024008945A (ja) | 膵島細胞の増殖を促進する方法 | |
EP2773668B1 (en) | Pulmonary hypertension | |
US20090004182A1 (en) | Methods to Treat or Prevent Viral-Associated Lymphoproliferative Disorders | |
JP2024509702A (ja) | 急性covid-19および急性後covid-19を発症するリスクを評価するためのバイオマーカー | |
US20220267428A1 (en) | BLOCKADE OF RGMb FOR TREATING INFLAMMATORY BOWEL DISEASE AND COLITIS | |
JP2005516907A (ja) | 霊長類ifn−ガンマ結合分子の使用 | |
KR20150090107A (ko) | 중피종의 치료 방법 | |
EP3389704B1 (en) | Vaccination against diabetes, obesity and complications thereof | |
KR20230142834A (ko) | 항-cd38 항체 및 이의 용도 | |
WO2024018440A1 (en) | Neutralization of acyl-coa binding protein for the treatment of cardiac dysfunction | |
WO2023168087A1 (en) | Methods and compositions for treating and preventing fibrosis | |
JP2024507986A (ja) | 抗体関連型移植拒絶反応の治療に使用するための抗cd38抗体 | |
JP2023526091A (ja) | がんの予防または治療用薬学的組成物 | |
CN117561078A (zh) | 用于评价发展急性covid-19和急性后covid-19的风险的生物标志物 | |
Bilinski | Evaluation of UNK Cell Capacity to Initiate Pregnancy-associated Spiral Artery Remodelling |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: CYTOKINE PHARMASCIENCES, INC., PENNSYLVANIA Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:GIROIR, BRETT P.;WILLIS, MONTE S.;DE LA CRUZ JR., VIDAL F;AND OTHERS;REEL/FRAME:016654/0228;SIGNING DATES FROM 20050419 TO 20050527 Owner name: BOARD OF REGENTS, THE UNIVERSITY OF TEXAS SYSTEM, Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:GIROIR, BRETT P.;WILLIS, MONTE S.;DE LA CRUZ JR., VIDAL F;AND OTHERS;REEL/FRAME:016654/0228;SIGNING DATES FROM 20050419 TO 20050527 |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |
|
AS | Assignment |
Owner name: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF Free format text: CONFIRMATORY LICENSE;ASSIGNOR:UNIVERSITY OF TEXAS SW MEDICAL CENTER AT DALLAS;REEL/FRAME:021700/0005 Effective date: 20040922 |