ZA200401552B - Osteogenic growth oligopeptides as stimulants of hematopoiesis. - Google Patents
Osteogenic growth oligopeptides as stimulants of hematopoiesis. Download PDFInfo
- Publication number
- ZA200401552B ZA200401552B ZA200401552A ZA200401552A ZA200401552B ZA 200401552 B ZA200401552 B ZA 200401552B ZA 200401552 A ZA200401552 A ZA 200401552A ZA 200401552 A ZA200401552 A ZA 200401552A ZA 200401552 B ZA200401552 B ZA 200401552B
- Authority
- ZA
- South Africa
- Prior art keywords
- gly
- phe
- tyr
- composition
- substance
- Prior art date
Links
- 108010038807 Oligopeptides Proteins 0.000 title claims description 102
- 102000015636 Oligopeptides Human genes 0.000 title claims description 102
- 230000011132 hemopoiesis Effects 0.000 title description 18
- 230000002188 osteogenic effect Effects 0.000 title description 8
- 230000012010 growth Effects 0.000 title description 6
- 239000000021 stimulant Substances 0.000 title description 2
- 210000004027 cell Anatomy 0.000 claims description 130
- 108010034423 historphin Proteins 0.000 claims description 100
- 210000001185 bone marrow Anatomy 0.000 claims description 99
- 210000000130 stem cell Anatomy 0.000 claims description 98
- 238000000034 method Methods 0.000 claims description 94
- 239000000203 mixture Substances 0.000 claims description 70
- 210000003958 hematopoietic stem cell Anatomy 0.000 claims description 66
- 239000008194 pharmaceutical composition Substances 0.000 claims description 58
- 208000003476 primary myelofibrosis Diseases 0.000 claims description 49
- 210000004369 blood Anatomy 0.000 claims description 48
- 239000008280 blood Substances 0.000 claims description 48
- 230000003394 haemopoietic effect Effects 0.000 claims description 45
- 239000000126 substance Substances 0.000 claims description 39
- 238000011282 treatment Methods 0.000 claims description 39
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 claims description 35
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 claims description 35
- 238000010322 bone marrow transplantation Methods 0.000 claims description 33
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 29
- 230000002708 enhancing effect Effects 0.000 claims description 26
- 230000001965 increasing effect Effects 0.000 claims description 26
- 238000002360 preparation method Methods 0.000 claims description 25
- BGFPKYKDLYYTJH-OALUTQOASA-N 2-[[2-[[(2s)-2-[[2-[[(2s)-2-amino-3-(4-hydroxyphenyl)propanoyl]amino]acetyl]amino]-3-phenylpropanoyl]amino]acetyl]amino]acetic acid Chemical compound C([C@H](N)C(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)NCC(=O)NCC(O)=O)C1=CC=C(O)C=C1 BGFPKYKDLYYTJH-OALUTQOASA-N 0.000 claims description 23
- 230000035755 proliferation Effects 0.000 claims description 21
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 20
- 238000002512 chemotherapy Methods 0.000 claims description 19
- 208000014951 hematologic disease Diseases 0.000 claims description 17
- 238000000338 in vitro Methods 0.000 claims description 17
- 241000124008 Mammalia Species 0.000 claims description 16
- VJLLEKDQJSMHRU-STQMWFEESA-N Phe-Gly-Met Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CCSC)C(O)=O VJLLEKDQJSMHRU-STQMWFEESA-N 0.000 claims description 16
- 208000014767 Myeloproliferative disease Diseases 0.000 claims description 15
- 206010028980 Neoplasm Diseases 0.000 claims description 15
- 210000005259 peripheral blood Anatomy 0.000 claims description 15
- 239000011886 peripheral blood Substances 0.000 claims description 15
- 102000004269 Granulocyte Colony-Stimulating Factor Human genes 0.000 claims description 14
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 claims description 14
- 210000002960 bfu-e Anatomy 0.000 claims description 13
- 230000001332 colony forming effect Effects 0.000 claims description 12
- 208000032467 Aplastic anaemia Diseases 0.000 claims description 11
- 102000004127 Cytokines Human genes 0.000 claims description 11
- 108090000695 Cytokines Proteins 0.000 claims description 11
- 239000004615 ingredient Substances 0.000 claims description 11
- 208000032839 leukemia Diseases 0.000 claims description 10
- 208000017604 Hodgkin disease Diseases 0.000 claims description 9
- 208000010747 Hodgkins lymphoma Diseases 0.000 claims description 9
- 206010025323 Lymphomas Diseases 0.000 claims description 9
- 208000026278 immune system disease Diseases 0.000 claims description 9
- 210000001616 monocyte Anatomy 0.000 claims description 9
- 238000011084 recovery Methods 0.000 claims description 9
- 238000002560 therapeutic procedure Methods 0.000 claims description 9
- 102000007651 Macrophage Colony-Stimulating Factor Human genes 0.000 claims description 7
- 108010046938 Macrophage Colony-Stimulating Factor Proteins 0.000 claims description 7
- 201000007224 Myeloproliferative neoplasm Diseases 0.000 claims description 7
- 238000011773 genetically engineered mouse model Methods 0.000 claims description 7
- 101100335081 Mus musculus Flt3 gene Proteins 0.000 claims description 6
- 230000003247 decreasing effect Effects 0.000 claims description 6
- 210000003714 granulocyte Anatomy 0.000 claims description 6
- 102000013925 CD34 antigen Human genes 0.000 claims description 5
- 108050003733 CD34 antigen Proteins 0.000 claims description 5
- 102000004889 Interleukin-6 Human genes 0.000 claims description 5
- 108090001005 Interleukin-6 Proteins 0.000 claims description 5
- 102000003951 Erythropoietin Human genes 0.000 claims description 4
- 108090000394 Erythropoietin Proteins 0.000 claims description 4
- 108010002352 Interleukin-1 Proteins 0.000 claims description 4
- 102000000589 Interleukin-1 Human genes 0.000 claims description 4
- 102000004058 Leukemia inhibitory factor Human genes 0.000 claims description 4
- 108090000581 Leukemia inhibitory factor Proteins 0.000 claims description 4
- 102000036693 Thrombopoietin Human genes 0.000 claims description 4
- 108010041111 Thrombopoietin Proteins 0.000 claims description 4
- 238000004820 blood count Methods 0.000 claims description 4
- 229940105423 erythropoietin Drugs 0.000 claims description 4
- 210000004976 peripheral blood cell Anatomy 0.000 claims description 4
- OXCMYAYHXIHQOA-UHFFFAOYSA-N potassium;[2-butyl-5-chloro-3-[[4-[2-(1,2,4-triaza-3-azanidacyclopenta-1,4-dien-5-yl)phenyl]phenyl]methyl]imidazol-4-yl]methanol Chemical compound [K+].CCCCC1=NC(Cl)=C(CO)N1CC1=CC=C(C=2C(=CC=CC=2)C2=N[N-]N=N2)C=C1 OXCMYAYHXIHQOA-UHFFFAOYSA-N 0.000 claims description 4
- 108090000978 Interleukin-4 Proteins 0.000 claims description 3
- 102000004388 Interleukin-4 Human genes 0.000 claims description 3
- 108010002616 Interleukin-5 Proteins 0.000 claims description 3
- 102000015215 Stem Cell Factor Human genes 0.000 claims description 3
- 108010039445 Stem Cell Factor Proteins 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 3
- 108090000174 Interleukin-10 Proteins 0.000 claims description 2
- 102000000743 Interleukin-5 Human genes 0.000 claims description 2
- 108010002586 Interleukin-7 Proteins 0.000 claims description 2
- 108090001007 Interleukin-8 Proteins 0.000 claims description 2
- 108010002335 Interleukin-9 Proteins 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 17
- 108010065805 Interleukin-12 Proteins 0.000 claims 1
- 108010002386 Interleukin-3 Proteins 0.000 claims 1
- 230000002265 prevention Effects 0.000 claims 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims 1
- BGFPKYKDLYYTJH-UHFFFAOYSA-N 2-[[2-[[2-[[2-[[2-amino-3-(4-hydroxyphenyl)propanoyl]amino]acetyl]amino]-3-phenylpropanoyl]amino]acetyl]amino]acetic acid Chemical compound C=1C=CC=CC=1CC(C(=O)NCC(=O)NCC(O)=O)NC(=O)CNC(=O)C(N)CC1=CC=C(O)C=C1 BGFPKYKDLYYTJH-UHFFFAOYSA-N 0.000 description 76
- 241000699670 Mus sp. Species 0.000 description 39
- 230000000694 effects Effects 0.000 description 37
- VNTJGCYVIRTGMZ-PXGLAOGESA-N 2-[[2-[[(2s)-2-[[2-[[(2s)-2-[[(2s)-2-[[(2s,3r)-2-[[(2s)-2-[[2-[[(2s)-5-amino-2-[[(2s)-2-[[(2s)-6-amino-2-[[(2s)-2-[[(2s)-2-aminopropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]-5-oxopentanoyl]amino]acety Chemical compound C([C@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@H](C)N)CC(C)C)C(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)NCC(=O)NCC(O)=O)C1=CC=C(O)C=C1 VNTJGCYVIRTGMZ-PXGLAOGESA-N 0.000 description 24
- 101800003595 Osteogenic growth peptide Proteins 0.000 description 24
- 210000000265 leukocyte Anatomy 0.000 description 24
- 210000000601 blood cell Anatomy 0.000 description 23
- 230000004936 stimulating effect Effects 0.000 description 15
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 14
- 238000001727 in vivo Methods 0.000 description 14
- 239000002953 phosphate buffered saline Substances 0.000 description 14
- 239000003981 vehicle Substances 0.000 description 13
- 241001465754 Metazoa Species 0.000 description 12
- 241000699666 Mus <mouse, genus> Species 0.000 description 12
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 12
- 229920001184 polypeptide Polymers 0.000 description 12
- 230000001225 therapeutic effect Effects 0.000 description 12
- 210000000988 bone and bone Anatomy 0.000 description 11
- 238000004519 manufacturing process Methods 0.000 description 11
- 239000002243 precursor Substances 0.000 description 11
- 210000001519 tissue Anatomy 0.000 description 11
- 210000003743 erythrocyte Anatomy 0.000 description 10
- 238000002054 transplantation Methods 0.000 description 9
- 239000000969 carrier Substances 0.000 description 8
- 229960004397 cyclophosphamide Drugs 0.000 description 8
- 230000004069 differentiation Effects 0.000 description 8
- 201000010099 disease Diseases 0.000 description 8
- 238000002347 injection Methods 0.000 description 8
- 239000007924 injection Substances 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 7
- 230000015572 biosynthetic process Effects 0.000 description 7
- 210000001772 blood platelet Anatomy 0.000 description 7
- 238000012423 maintenance Methods 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 230000008093 supporting effect Effects 0.000 description 7
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 6
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 6
- 238000003556 assay Methods 0.000 description 6
- 239000003937 drug carrier Substances 0.000 description 6
- 230000000925 erythroid effect Effects 0.000 description 6
- 230000007774 longterm Effects 0.000 description 6
- 210000004698 lymphocyte Anatomy 0.000 description 6
- 210000002540 macrophage Anatomy 0.000 description 6
- 206010028537 myelofibrosis Diseases 0.000 description 6
- 210000003643 myeloid progenitor cell Anatomy 0.000 description 6
- 210000000963 osteoblast Anatomy 0.000 description 6
- 239000000546 pharmaceutical excipient Substances 0.000 description 6
- 238000001959 radiotherapy Methods 0.000 description 6
- 229940024606 amino acid Drugs 0.000 description 5
- 150000001413 amino acids Chemical group 0.000 description 5
- 210000002798 bone marrow cell Anatomy 0.000 description 5
- 210000004899 c-terminal region Anatomy 0.000 description 5
- 231100000673 dose–response relationship Toxicity 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 230000010437 erythropoiesis Effects 0.000 description 5
- 239000003102 growth factor Substances 0.000 description 5
- 230000002489 hematologic effect Effects 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 239000003094 microcapsule Substances 0.000 description 5
- 239000003381 stabilizer Substances 0.000 description 5
- 210000002536 stromal cell Anatomy 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 206010016654 Fibrosis Diseases 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- 241001529936 Murinae Species 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 238000001574 biopsy Methods 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 4
- 208000035475 disorder Diseases 0.000 description 4
- 230000004761 fibrosis Effects 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 230000003993 interaction Effects 0.000 description 4
- 229940100601 interleukin-6 Drugs 0.000 description 4
- 210000003593 megakaryocyte Anatomy 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000007920 subcutaneous administration Methods 0.000 description 4
- 231100000419 toxicity Toxicity 0.000 description 4
- 230000001988 toxicity Effects 0.000 description 4
- 206010000830 Acute leukaemia Diseases 0.000 description 3
- 102000001554 Hemoglobins Human genes 0.000 description 3
- 108010054147 Hemoglobins Proteins 0.000 description 3
- -1 IL-3 Proteins 0.000 description 3
- 238000002679 ablation Methods 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 208000007502 anemia Diseases 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 230000004663 cell proliferation Effects 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 239000000470 constituent Substances 0.000 description 3
- 229920001577 copolymer Polymers 0.000 description 3
- 238000003745 diagnosis Methods 0.000 description 3
- 210000000267 erythroid cell Anatomy 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 210000004700 fetal blood Anatomy 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 210000002360 granulocyte-macrophage progenitor cell Anatomy 0.000 description 3
- 230000002607 hemopoietic effect Effects 0.000 description 3
- 238000001990 intravenous administration Methods 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 210000000066 myeloid cell Anatomy 0.000 description 3
- 210000000440 neutrophil Anatomy 0.000 description 3
- 231100000252 nontoxic Toxicity 0.000 description 3
- 230000003000 nontoxic effect Effects 0.000 description 3
- 230000011164 ossification Effects 0.000 description 3
- 210000005009 osteogenic cell Anatomy 0.000 description 3
- 229920000642 polymer Polymers 0.000 description 3
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 3
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 3
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 3
- 230000005855 radiation Effects 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 210000003660 reticulum Anatomy 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 239000008227 sterile water for injection Substances 0.000 description 3
- 238000013268 sustained release Methods 0.000 description 3
- 239000012730 sustained-release form Substances 0.000 description 3
- 230000001052 transient effect Effects 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- 206010002961 Aplasia Diseases 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 description 2
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 101000980898 Homo sapiens Cell division cycle-associated protein 4 Proteins 0.000 description 2
- 101000987586 Homo sapiens Eosinophil peroxidase Proteins 0.000 description 2
- 101000920686 Homo sapiens Erythropoietin Proteins 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- 102000006992 Interferon-alpha Human genes 0.000 description 2
- 108010047761 Interferon-alpha Proteins 0.000 description 2
- 108010000817 Leuprolide Proteins 0.000 description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 2
- 101710160107 Outer membrane protein A Proteins 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 206010033661 Pancytopenia Diseases 0.000 description 2
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 description 2
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 description 2
- RADKZDMFGJYCBB-UHFFFAOYSA-N Pyridoxal Chemical compound CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 description 2
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 2
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 2
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 2
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 2
- 230000000735 allogeneic effect Effects 0.000 description 2
- APKFDSVGJQXUKY-INPOYWNPSA-N amphotericin B Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/C=C/C=C/C=C/[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 APKFDSVGJQXUKY-INPOYWNPSA-N 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 210000003719 b-lymphocyte Anatomy 0.000 description 2
- 210000003651 basophil Anatomy 0.000 description 2
- 102000023732 binding proteins Human genes 0.000 description 2
- 108091008324 binding proteins Proteins 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- DQXBYHZEEUGOBF-UHFFFAOYSA-N but-3-enoic acid;ethene Chemical compound C=C.OC(=O)CC=C DQXBYHZEEUGOBF-UHFFFAOYSA-N 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 230000005757 colony formation Effects 0.000 description 2
- 239000003405 delayed action preparation Substances 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 210000003013 erythroid precursor cell Anatomy 0.000 description 2
- 239000005038 ethylene vinyl acetate Substances 0.000 description 2
- 210000002950 fibroblast Anatomy 0.000 description 2
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 2
- 208000018706 hematopoietic system disease Diseases 0.000 description 2
- 102000044493 human CDCA4 Human genes 0.000 description 2
- 102000044890 human EPO Human genes 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- 239000000017 hydrogel Substances 0.000 description 2
- 230000005847 immunogenicity Effects 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 description 2
- 230000002297 mitogenic effect Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 239000002088 nanocapsule Substances 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 238000010647 peptide synthesis reaction Methods 0.000 description 2
- 235000021317 phosphate Nutrition 0.000 description 2
- 239000000902 placebo Substances 0.000 description 2
- 229940068196 placebo Drugs 0.000 description 2
- 229920001200 poly(ethylene-vinyl acetate) Polymers 0.000 description 2
- 230000002062 proliferating effect Effects 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 238000000611 regression analysis Methods 0.000 description 2
- 235000002639 sodium chloride Nutrition 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000006641 stabilisation Effects 0.000 description 2
- 238000011105 stabilization Methods 0.000 description 2
- 238000010254 subcutaneous injection Methods 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 230000009885 systemic effect Effects 0.000 description 2
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)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)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 2
- 238000011287 therapeutic dose Methods 0.000 description 2
- 230000000699 topical effect Effects 0.000 description 2
- 102000003390 tumor necrosis factor Human genes 0.000 description 2
- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 description 1
- FSPQCTGGIANIJZ-UHFFFAOYSA-N 2-[[(3,4-dimethoxyphenyl)-oxomethyl]amino]-4,5,6,7-tetrahydro-1-benzothiophene-3-carboxamide Chemical compound C1=C(OC)C(OC)=CC=C1C(=O)NC1=C(C(N)=O)C(CCCC2)=C2S1 FSPQCTGGIANIJZ-UHFFFAOYSA-N 0.000 description 1
- XBBVURRQGJPTHH-UHFFFAOYSA-N 2-hydroxyacetic acid;2-hydroxypropanoic acid Chemical compound OCC(O)=O.CC(O)C(O)=O XBBVURRQGJPTHH-UHFFFAOYSA-N 0.000 description 1
- 208000030507 AIDS Diseases 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 206010065553 Bone marrow failure Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 238000010599 BrdU assay Methods 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- COVZYZSDYWQREU-UHFFFAOYSA-N Busulfan Chemical compound CS(=O)(=O)OCCCCOS(C)(=O)=O COVZYZSDYWQREU-UHFFFAOYSA-N 0.000 description 1
- 102000000584 Calmodulin Human genes 0.000 description 1
- 108010041952 Calmodulin Proteins 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 208000010833 Chronic myeloid leukaemia Diseases 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 102000007644 Colony-Stimulating Factors Human genes 0.000 description 1
- 108010071942 Colony-Stimulating Factors Proteins 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
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 description 1
- 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 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
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 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 1
- 241000283086 Equidae Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 206010017533 Fungal infection Diseases 0.000 description 1
- 108091006027 G proteins Proteins 0.000 description 1
- 102000030782 GTP binding Human genes 0.000 description 1
- 108091000058 GTP-Binding Proteins 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- 208000002250 Hematologic Neoplasms Diseases 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 101100220044 Homo sapiens CD34 gene Proteins 0.000 description 1
- 101000716729 Homo sapiens Kit ligand Proteins 0.000 description 1
- 108091006905 Human Serum Albumin Proteins 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- VSNHCAURESNICA-UHFFFAOYSA-N Hydroxyurea Chemical compound NC(=O)NO VSNHCAURESNICA-UHFFFAOYSA-N 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
- 238000012695 Interfacial polymerization Methods 0.000 description 1
- 108010002350 Interleukin-2 Proteins 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 102000043136 MAP kinase family Human genes 0.000 description 1
- 108091054455 MAP kinase family Proteins 0.000 description 1
- 239000004907 Macro-emulsion Substances 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 208000031888 Mycoses Diseases 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- 241000906034 Orthops Species 0.000 description 1
- 102000004067 Osteocalcin Human genes 0.000 description 1
- 108090000573 Osteocalcin Proteins 0.000 description 1
- 208000031951 Primary immunodeficiency Diseases 0.000 description 1
- 108010007568 Protamines Proteins 0.000 description 1
- 102000007327 Protamines Human genes 0.000 description 1
- 102100020718 Receptor-type tyrosine-protein kinase FLT3 Human genes 0.000 description 1
- 101710151245 Receptor-type tyrosine-protein kinase FLT3 Proteins 0.000 description 1
- 206010039710 Scleroderma Diseases 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 206010041660 Splenomegaly Diseases 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- DFPAKSUCGFBDDF-ZQBYOMGUSA-N [14c]-nicotinamide Chemical compound N[14C](=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-ZQBYOMGUSA-N 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- CEGOLXSVJUTHNZ-UHFFFAOYSA-K aluminium tristearate Chemical compound [Al+3].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O CEGOLXSVJUTHNZ-UHFFFAOYSA-K 0.000 description 1
- 229940063655 aluminum stearate Drugs 0.000 description 1
- 238000001949 anaesthesia Methods 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000000202 analgesic effect Effects 0.000 description 1
- 239000003098 androgen Substances 0.000 description 1
- 229940030486 androgens Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 230000001363 autoimmune Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 210000003995 blood forming stem cell Anatomy 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 238000006664 bond formation reaction Methods 0.000 description 1
- 210000002449 bone cell Anatomy 0.000 description 1
- 210000004271 bone marrow stromal cell Anatomy 0.000 description 1
- 230000010478 bone regeneration Effects 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- 229960002092 busulfan Drugs 0.000 description 1
- 230000002308 calcification Effects 0.000 description 1
- FAPWYRCQGJNNSJ-UBKPKTQASA-L calcium D-pantothenic acid Chemical compound [Ca+2].OCC(C)(C)[C@@H](O)C(=O)NCCC([O-])=O.OCC(C)(C)[C@@H](O)C(=O)NCCC([O-])=O FAPWYRCQGJNNSJ-UBKPKTQASA-L 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 239000004359 castor oil Substances 0.000 description 1
- 235000019438 castor oil Nutrition 0.000 description 1
- 238000010370 cell cloning Methods 0.000 description 1
- 230000011712 cell development Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000036978 cell physiology Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 238000001311 chemical methods and process Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000003021 clonogenic effect Effects 0.000 description 1
- 238000005354 coacervation Methods 0.000 description 1
- 239000008119 colloidal silica Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000016396 cytokine production Effects 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000002435 cytoreductive effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- GXGAKHNRMVGRPK-UHFFFAOYSA-N dimagnesium;dioxido-bis[[oxido(oxo)silyl]oxy]silane Chemical compound [Mg+2].[Mg+2].[O-][Si](=O)O[Si]([O-])([O-])O[Si]([O-])=O GXGAKHNRMVGRPK-UHFFFAOYSA-N 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 235000019797 dipotassium phosphate Nutrition 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 230000007783 downstream signaling Effects 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000003979 eosinophil Anatomy 0.000 description 1
- 230000008029 eradication Effects 0.000 description 1
- BEFDCLMNVWHSGT-UHFFFAOYSA-N ethenylcyclopentane Chemical compound C=CC1CCCC1 BEFDCLMNVWHSGT-UHFFFAOYSA-N 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 230000003328 fibroblastic effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 238000012637 gene transfection Methods 0.000 description 1
- CEAZRRDELHUEMR-UHFFFAOYSA-N gentamicin Chemical compound O1C(C(C)NC)CCC(N)C1OC1C(O)C(OC2C(C(NC)C(C)(O)CO2)O)C(N)CC1N CEAZRRDELHUEMR-UHFFFAOYSA-N 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 125000005456 glyceride group Chemical group 0.000 description 1
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 1
- 229960002449 glycine Drugs 0.000 description 1
- 229940093915 gynecological organic acid Drugs 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 210000003630 histaminocyte Anatomy 0.000 description 1
- 102000055151 human KITLG Human genes 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 229920001477 hydrophilic polymer Polymers 0.000 description 1
- 229920001600 hydrophobic polymer Polymers 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 229960001330 hydroxycarbamide Drugs 0.000 description 1
- 229920003063 hydroxymethyl cellulose Polymers 0.000 description 1
- 229940031574 hydroxymethyl cellulose Drugs 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 230000003463 hyperproliferative effect Effects 0.000 description 1
- 230000000642 iatrogenic effect Effects 0.000 description 1
- 210000001621 ilium bone Anatomy 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000017555 immunoglobulin mediated immune response Effects 0.000 description 1
- 229940072221 immunoglobulins Drugs 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 238000005470 impregnation Methods 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000007919 intrasynovial administration Methods 0.000 description 1
- 238000007913 intrathecal administration Methods 0.000 description 1
- 230000002601 intratumoral effect Effects 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 230000021633 leukocyte mediated immunity Effects 0.000 description 1
- GFIJNRVAKGFPGQ-LIJARHBVSA-N leuprolide Chemical compound CCNC(=O)[C@@H]1CCCN1C(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H]1NC(=O)CC1)CC1=CC=C(O)C=C1 GFIJNRVAKGFPGQ-LIJARHBVSA-N 0.000 description 1
- RGLRXNKKBLIBQS-XNHQSDQCSA-N leuprolide acetate Chemical compound CC(O)=O.CCNC(=O)[C@@H]1CCCN1C(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H]1NC(=O)CC1)CC1=CC=C(O)C=C1 RGLRXNKKBLIBQS-XNHQSDQCSA-N 0.000 description 1
- 229960004338 leuprorelin Drugs 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 238000005461 lubrication Methods 0.000 description 1
- 229940087857 lupron Drugs 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 239000000391 magnesium silicate Substances 0.000 description 1
- 229940099273 magnesium trisilicate Drugs 0.000 description 1
- 229910000386 magnesium trisilicate Inorganic materials 0.000 description 1
- 235000019793 magnesium trisilicate Nutrition 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 239000004530 micro-emulsion Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 239000003226 mitogen Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000001483 mobilizing effect Effects 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 229940126619 mouse monoclonal antibody Drugs 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000001400 myeloablative effect Effects 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 239000002736 nonionic surfactant Substances 0.000 description 1
- 229940127240 opiate Drugs 0.000 description 1
- 210000004789 organ system Anatomy 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 230000001582 osteoblastic effect Effects 0.000 description 1
- 210000002997 osteoclast Anatomy 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 229940127557 pharmaceutical product Drugs 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 210000001778 pluripotent stem cell Anatomy 0.000 description 1
- 229920000747 poly(lactic acid) Polymers 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 231100000683 possible toxicity Toxicity 0.000 description 1
- 235000010241 potassium sorbate Nutrition 0.000 description 1
- 239000004302 potassium sorbate Substances 0.000 description 1
- 229940069338 potassium sorbate Drugs 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 238000009101 premedication Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 229950008679 protamine sulfate Drugs 0.000 description 1
- 230000006337 proteolytic cleavage Effects 0.000 description 1
- 229960003581 pyridoxal Drugs 0.000 description 1
- 235000008164 pyridoxal Nutrition 0.000 description 1
- 239000011674 pyridoxal Substances 0.000 description 1
- 238000011127 radiochemotherapy Methods 0.000 description 1
- 230000008707 rearrangement Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 230000002629 repopulating effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000010242 retro-orbital bleeding Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 229960002477 riboflavin Drugs 0.000 description 1
- 235000019192 riboflavin Nutrition 0.000 description 1
- 239000002151 riboflavin Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 230000000405 serological effect Effects 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 235000010199 sorbic acid Nutrition 0.000 description 1
- 229940075582 sorbic acid Drugs 0.000 description 1
- 239000004334 sorbic acid Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 230000003393 splenic effect Effects 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 230000023895 stem cell maintenance Effects 0.000 description 1
- 238000011146 sterile filtration Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 239000006190 sub-lingual tablet Substances 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 230000003319 supportive effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 206010043554 thrombocytopenia Diseases 0.000 description 1
- 230000003582 thrombocytopenic effect Effects 0.000 description 1
- 229940104230 thymidine Drugs 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 230000036962 time dependent Effects 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 235000019786 weight gain Nutrition 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
- 229920001285 xanthan gum Polymers 0.000 description 1
- 150000003751 zinc Chemical class 0.000 description 1
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Description
OSTEOGENIC GROWTH OLIGOPEPTIDES AS STIMULLANTS OF HEMATOPOIESIS
The present invention relates to the use of oligopeptides corresponding to the C-terminal portion of OGP, as stimulators of hematopoiesis. More specifically, these oligopeptides enhance engraftment of bone marrow transplants, hematopoietic reconstruction, bone marrow re-population and number of circulating stem cells, particularly after chemotherapy or irradiation. The invention further provides methods for using these oligopeptides and pharmaceutical compositions comprising them.
Biological and biochemical interactions between bone and bone marrow are far from being fully understood. However recent studies confirm the ~ role of bone marrow derived osteogenic cells in supporting hematopoietic cell development [Teichman, R.S., et al., Hematol. 4:421-426 (2000)].
Bone marrow transplantation studies confirm the bi-directional interactions between the two systems. Bone marrow ablation or radiation damage triggers an initial local, transient osteogenic reaction [Amsel, S,, et al., Anat. Rec. 164:101-111 (1969); Patt, H.M., and Maloney,
M.A., Exp. Hematol. 3:135-148 (1975)]. In this osteogenic phase, trabeculae are formed in the marrow cavity. The trabeculae are transient and are resorbed during the reconstitution of haematopoietic marrow.
Moreover, in human bone marrow donors, an increase in serum bone formation markers osteocalcin and alkaline phosphatase was recorded . after the removal of a substantial portion of iliac bone marrow [Foldes, J . et al., J. Bone Miner. Res. 4:643-646 (1989)]. The hypothesis that human osteoblasts support human hematopoietic progenitor cells is very intriguing: these cells produce factors that directly stimulate the formation of hematopoietic colonies without the addition of exogenously supplied growth factors. In fact, osteoblasts secrete several cytokines including granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor (TNF), and interleukin 6 (IL-6). Besides, cultured A osteoblasts support the maintenance of immature phenotype in hematopoietic stem cells [Taichman, et al., Blood 87:518-524 (1996)]. ]
Several of these growth factors improve in vivo bone marrow re-population and peripheral stem cell mobilization after high dose chemotherapy. Among them, G-CSF, GM-CSF, IL-3 (Interleukine-3) and
SCF (Stem Cell Factor) have been extensively evaluated [Bungart, B., et al., Br. J. Haematol. 76:174 (1990); Lant, T., et al., Blood 85:275 (1995);
Brugger, W., et al., Blood 79:1193 (1992); Molinex, G., et al., Blood 78:961 (1991)] and many others, such as FLT-3, are under study for clinical use [Ashihara, E., et al., Europ. J. Haematol. 60:86 (1998)]. Advances in this field in recent years, have allowed an understanding of several physiological aspects of bone marrow function. Moreover, the ability to modulate differentiation and proliferation of haematological precursors is at the basis of the more innovative therapies such as peripheral blood stem cell transplant, gene transfection and ex vivo expansion of stem cells. In spite of this impressive progress, several aspects of stem cell physiology have not been fully clarified, and several factors, both soluble or cell membrane related, are suspected of being involved in the physiological or pathological proliferation/differentiation of bone marrow cells. The increasing number of agents shown to be able to regulate hematopoiesis supports the critical question regarding the redundancy or subtlety of hematopoietic regulators [Metcaff, D., et al., Blood 82:3515 (1993)]. :
In addition to the role of classically defined growth factors, several biological agents and cell types could improve or modify both in vivo and ex vivo therapeutic strategies. Human bone marrow-derived endothelial cells support long term proliferation and differentiation of myeloid and megakaryocytic progenitors [Rafii, S., et al., Blood 86:3353 (1995)]; accessory cells may support hematological recovery after bone marrow ‘ transplant [Bonnet, D., et al., Bone Marrow Transpl. 23:203 (1991); and, even more interesting for present purposes, osteoblasts may enhance the engraftment after HLA-unrelated bone marrow transplant in mice (El-Badri, N.S, et al., Exp. Hematol. 26:110 (1998)].
Many chemical structures have been investigated in order to assess a possible role in bone marrow physiology. For example, effects of glycosaminoglycans have been evaluated both on leukemia-derived cells lines [Volpi, N., et al., Exp. Cell Res. 215:119 (1994)] and in clonogenic tests on human cord blood-derived stem cells [Da Prato, I., et al., Leuk.
Res. 23:1015 (1999)]. Even short peptides have been synthesized to reach hemoregulatory and multilineage effects, possibly by enhancement of cytokine production by stromal cells [King, A.G., et al., Exp. Hematol. 20(4):531 (1992); Pelus, L.M,, et al., Exp. Hematol. 22:239 (1994)].
Idiopathic myelofibrosis (IMF) is the least common and carries the worst prognosis of the chronic myeloproliferative disorders. The primary pathogenic process is a clonal hematopoietic stem cell disorder which results in anemia, atypical megakaryocyte hyperplasia, splenomegaly and varying degrees of extramedullary hematopoiesis. In contrast, the characteristic stromal proliferation is a reactive phenomenon, resulting from the inappropriate release of megakaryocyte/platelet-derived growth factors, including platelet-derived growth factor (PDGF), transforming growth factor-beta (TGF-beta), basic fibroblast growth factor (bFGF), . epidermal growth factor (EGF), and calmodulin [Groopman, J., Ann.
Intern. Med. 92:857-858 (1980); Chvapil, M., Life Sci. 16:1345-1361 (1975)]. : The median survival of IMF patients is approximately 4 years. Therapeutic strategies in IMF remain predominantly supportive and directed towards the alleviation of symptoms and improvement in quality of life. The most common are blood transfusions, androgens and cytoreductive agents such as hydroxyurea. Bone marrow transplantation is increasingly being taken into consideration, but it still has to be regarded as an experimental approach. Interferon-alpha (IFN-alpha) has shown promising results in early hyperproliferative stages of IMF but has no or only very little effect in more advanced stages of the disease.
It has been previously shown by some of the inventors that osteogenic growth peptide (OGP), a 14-amino acid, highly conserved H4 histone-related peptide, increases blood and bone marrow cellularity and enhances engraftment of bone marrow transplants in mice [Bab, L.A,
Clin. Orthop. 313:64 (1995); Gurevitch, O., et al., Blood 88:4719 (1996) and US Patent No. 5,461,034]. OGP has been isolated from the osteogenic phast cf wost-ablotion hous niarrcw vogonecration Bab 1 stad,
Endocrinology, 128(5);2638 (1991)] and is physiologically present in high abundance in the blood, mainly as a complex with aj-macroglobulin (az-M) [Gavish, H., et al.,, Biochemistry, 36:14883-14888 (1997)].
Administered in vivo, it enhances bone formation and increases trabecular bone mass; in vitro, it stimulates the proliferation and alkaline phosphatase activity in osteogenic cell lines; in addition, it is mitogenic to fibroblasts [Greenberg, Z., et al., Biochim. Biophys. Acta. 1178:273 (1993)]. In addition to the activity of OGP on bone regeneration, osteoblast activation and fibroblast proliferation, it has been shown to induce, in vivo, a balanced increase in white blood cell (WBC) counts, and overall bone marrow cellularity in mice receiving myeloablative irradiation and syngeneic or semiallogeneic bone marrow transplants [Gurevitch, O., et al., ibid. (1996)].
The C-terminal pentapeptide of OGP, designated OGP(10-14), which seems to be generated by proteolytic cleavage of the full length OGP upon dissociation of the inactive complex with o,-M, is present in mammalian serum and osteogenic cell cultures at high levels [Bab, I., et al., J. Pept.
Res. 54:408 (1999)]. N-terminal modified OGP retains the OGP-like dose-dependent effect on cell proliferation, and it has been suggested that the carboxy-terminal pentapeptide is responsible for the binding to the ‘ putative OGP receptor [Greenberg, Z., et al., ibid. (1993)]. Additionally, the inventors have shown previously that in osteogenic MC3T3 E1 cells, mitogenic doses of OGP(10-14), but not OGP, enhance MAP kinase activity mm a time- and dose dependent manner. These findings indicate that the
OGP(10-14) is responsible for downstream signaling [Gabarin, et al., J. Cell
Biol. 81:594-603 (2001)]. It has further been shown that the active form of
OGP is its carboxy terminal pentapeptide OGP(10-14). Interestingly, the
OGP(10-14) does not form a complex with o,-M or other OGPBP (OGP binding protein) [Bab, I., J. Peptide Res. 54:408-414 (1999).
Therefore, the possible hematopoietic activity of synthetic oligopeptides analogous to the C-terminal region of native OGP was evaluated in the present invention. Some such osteogenically active specific peptides are described in US Patent No. 5,814,610. sOGP(10-14) has been described as having opiate and analgesic activities [Kharchenko et al., Vepr. Med.
Khim., 35(2) 106-109, (1989)].
Importantly, the present invention shows that previously known osteogenically active oligopeptides can act as stimulants of the hemopoietic system. For example, the synthetic OGP-derived pentapeptide designated OGP(10-14) has several properties such as increasing blood and bone marrow cellularity in mice, and enhancing engraftment of bone marrow transplants. This pentapeptide exhibited significant activity on peripheral blood cell recovery after . cyclophosphamide (CFA)- induced aplasia, and on stem cell mobilization.
Furthermore, the ex vivo effect of synthetic OGP(10-14) in bone marrow ) tissue samples from IMF patients was tested and demonstrated a substantial overall increase in the number of hematopoietic cells.
Moreover, the magnitude of the OGP (10-14) effect was directly related to the severity of IMF. These results indicate that OGP(10-14) may stimulate blood cell formation and rescue hematopoiesis.
It is therefore an object of the present invention to use OGP-drived v oligopeptides as hematopoietic growth factors. This and other objects of the invention will be elaborated on as the description proceeds.
In a first aspect, the invention relates to a pharmaceutical composition comprising as an effective ingredient at least one oligopeptide having stimulatory activity on the production of hematopoietic cells. The oligopeptide used according to the invention has a molecular weight of 200 to 1,000 Da aud may be an oligopeptide comprising any of the amino acid sequences Tyr-Gly-Phe-Gly-Gly, Tyr-Gly-Phe-His-Gly,
Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly. The pharmaceutical compositions of the invention optionally comprise a pharmaceutically acceptable carrier, diluent or excipient.
In one preferred embodiment of the present aspect, the pharmaceutical composition of the invention comprises an oligopeptide which is a pentapeptide having the formula: Tyr-Gly-Phe-Gly-Gly (designated
OGP(10-14)) and a pharmaceutically acceptable carrier.
In another embodiment, the pharmaceutical composition of the invention comprises an oligopeptide which is a pentapeptide having the formula:
Tyr-Gly-Phe-His-Gly.
In yet another embodiment, the pharmaceutical composition of the invention comprises an oligopeptide which is a tetrapeptide having the formula: Gly-Phe-Gly-Gly and a pharmaceutically acceptable carrier.
And in a further embodiment, the pharmaceutical composition of the 1mvention comprises an oligopeptide comprising the amino acid sequence
Met-Tyr-Gly-Phe-Gly-Gly and a pharmaceutically acceptable carrier, in . which the methionine residue is preferably acylated, namely an oligopeptide having the formula: Ac-Met-Tyr-Gly-Phe-Gly-Gly.
The pharmaceutical composition of the invention is intended for enhancement of engraftment of bone marrow transplants, hematopoietic reconstruction, bone marrow re-population and number of circulating stem cells.
In another embodiment the pharmaceutical composition of the invention 1s intended for enhancement of engraftment of bone marrow transplants, hematopoietic reconstruction, bone marrow re-population and number of circulating stem cells, particularly in patient receiving chemotherapy or irradiation.
The oligopeptide used. in the pharmaceutical composition of the invention increases the circulating multilineage progenitor cells percentage. These multilineage progenitor cells are the circulating early precursor CD34 positive cells.
Furthermore, the oligopeptide used as an effective ingredient in the pharmaceutical composition of the invention enhances the immature cell and monocyte recovery and selectively increases any one of the BFU-E and GEMM colony forming units (CFU). . The pharmaceutical composition of the invention is therefore intended for increasing the number of white blood cells (WBC), circulating : hematopoietic stem cells as well as overall bone marrow and blood cellularity.
In a specifically preferred embodiment, the composition of the invention is intended for supporting bone marrow transplantation. This effect is due to the activity of the oligopeptides on increasing the number of hematopoietic stem cells, accelerating the hematopoietic reconstruction N upon bone marrow transplantation and enhancing the overall cellularity of bone marrow. ’
According to another specifically preferred embodiment, the pharmaceutical composition of the invention is intended for use in treating bone marrow transplanted subjects suffering from hematological disorders, solid tumors, immunological disorders and/or aplastic anemia.
More specifically, the hematological disorders may be lymphomas, leukemias, Hodgkin's diseases and myeloproliferative disorders.
Particularly, the myeloproliferative disorder may be idiopathic myelofibrosis (IMF).
In a second aspect, the present invention relates to the use of an oligonucleotide comprising any one of the amino acid sequence
Tyr-Gly-Phe-Gly-Gly, Tyr-Gly-Phe-His-Gly, Gly-Phe-Gly-Gly and
Met-Tyr-Gly-Phe-Gly-Gly in the preparation of a pharmaceutical composition intended for enhancement of engraftment of a bone marrow transplant, hematopoietic reconstruction, bone marrow re-population and stimulating the number of circulating stem cells.
In a specific embodiment the oligonucleotides of the invention are used in the preparation of a pharmaceutical composition intended for enhancement of engraftment of a bone marrow transplant, hematopoietic reconstruction, bone marrow re-population and number of circulating - stem cells, particularly in patient receiving irradiation or chemotherapy .
According to a preferred embodiment, the above specific oligopeptides are used in the preparation of a pharmaceutical composition for increasing the number of circulating multilineage progenitor cells. These multilineage progenitor cells are the circulating early precursor CD34 positive cells.
Furthermore, the oligopeptides used in the preparation of the pharmaceutical composition of the invention enhance the immature cell, : monocyte recovery and selectively increase any one of the BFU-E and
GEMM colony forming units (CFU).
Accordingly, such oligopeptides may be used in the preparation of pharmaceutical composition intended for increasing the number of white blood cells (WBC), circulating hematopoietic stem cells, and/or overall bone marrow cellularity.
More specifically, the invention provides for the use of these oligopeptides in the preparation of a pharmaceutical composition for supporting bone marrow transplantation. This effect is due to the activity of the oligopeptides on increasing the number of stem cells, accelerating the hematopoietic reconstruction upon bone marrow transplantation and increasing the cellularity of bone marrow.
According to another specifically preferred embodiment, the present invention relates to the use of said oligopeptides in the preparation of a pharmaceutical composition which is intended for treating subjects suffering from hematological disorders, solid tumors, immunological disorders and/or aplastic anemia. More specifically, the hematological disorders may lymphomas, leukemias, Hodgkin's diseases or : myeloproliferative disorders, particularly idiopathic myelofibrosis (IMF). : In a third aspect, the present invention provides a method for enhancement of engraftment of a bone marrow transplant, hematopoietic reconstruction, bone marrow re-population and number of circulating stem cells. This method comprises the step of administering to a subject in need thereof, an effective amount of an oligopeptide having stimulatory activity on production of hematopoietic cells as described above, or of the composition of the invention. This method of the invention may be used . according to a preferred embodiment for enhancement of engraftment of a bone marrow transplant, hematopoietic reconstruction, bone marrow ’ re-population and number of circulating stem cells in a patient receiving irradiation or chemotherapy.
According to a specific embodiment of this aspect, the invention relates to a method of treating a subject suffering from a hematological disorder, solid tumor, immunological disorder or aplastic anemia. The method of the invention comprises administering to the subject a therapeutically effective amount of an oligopeptide having stimulatory activity on production of hematopoietic cells as described above, or of a composition comprising the same.
In another specific embodiment, this method can be used in support of the treatment of the subject by bone marrow transplantation.
More specifically, the hematological disorders may be lymphomas, leukemias, Hodgkin's disease or myeloproliferative disorders, particularly idiopathic myelofibrosis (IMF).
A preferred embodiment relates to a method for enhancing the number of hematopoietic stem/progenitor cells. According to the invention, this method comprises the steps of exposing these cells to an effective amount of an oligopeptide having stimulatory activity on production of . hematopoietic cells as described above, or to a composition comprising the same. )
In a specifically preferred embodiment, the method of the invention is intended for enhancing the proliferation of CD34 positive cells. ‘ In one specifically preferred embodiment, the cells are in cell culture and the method may be used ex vivo or in vitro.
Alternatively, the method of the invention may be used as an in vivo method of treatment, preferably of mammals, particularly humans.
The treated subject is one who suffers from, or is susceptible to, decreased blood cell levels, which may be caused by chemotherapy, irradiation therapy, or bone marrow transplantation therapy.
In yet another preferred embodiment, the invention relates to a method for in vitro or ex vivo maintaining and/or expanding hematopoietic stem cells present in a blood sample. This method comprises isolating peripheral blood cells from the blood sample, enriching blood progenitor cells expressing the CD34 antigen, cluttering the enriched blood progenitor cells under suitable conditions, and treating said cells with an oligopeptide having stimulatory activity on production of hematopoietic cells as described above or with a composition comprising the same.
In vivo treatment according to the invention relates to a method for re-populating blood cells in a mammal. This method comprises the steps of administering to said mammal a therapeutically effective amount of an oligopeptide having stimulatory activity on hematopoietic cells as described above, or of a composition comprising the same. These ] hematopoietic cells may be erythroid, myeloid or lymphoid cells.
Fig. 1 - A dose dependent effect of pretreatment with sOGP(10-14) on the total number of femoral marrow cells in mice after combined ablative radiotherapy/BMT }
OGP(10-14) at the indicated dose was daily injected subcutaneously for 12 days to female C57 BL mice. On day 8 after the onset of OGP(10-14) treatment the mice were subjected to 900 Rad X-ray irradiation, followed by intravenous administration of 105 syngeneic unselected bone marrow cells. On day 14 after the onset of treatment the mice were sacrificed and the femoral bone marrow washed out into phosphate buffered saline. A single cell suspension was prepared by drawing the preparation several times through graded syringe needles. Cell counts were carried out in a hemocytometer. C - control mice given phosphate buffered saline only.
Dala are eaii + SI ooutained in at 1east seven mice per condition.
Abbreviations: Fem (femoral), Marr C (marrow cells), D (day), mou (mouse), premed (premedication) stimu (stimulation) and cellu (cellularity).
Figs. 2A-C - OGP(10-14) stimulates blood cell counts in dose and time dependent manner in mice undergoing chemoablation of hematopoietic tissues
Male ICR mice weighing 25 gm each were subjected to chemoablation using cyclophosphamide (CFA), 5 mg/mouse, injected intraperitoneally on days 0 and 1, we injection each day. OGP(10-14) was dissolved in “sterile water for injection” and 0.1 ml of the indicated doses or water only (vehicle) was administered subcutaneously in the nape daily from day -7 to day —1 and from day +2 to day +8. Data are mean + SD obtained in 20 animals per condition. *: significant over CFA+vehicle, p<0.05; **: : significant over 1 nmol OGP(10-14) group, p<0.05.
Fig. 2A shows total white blood cell counts.
Fig. 2B shows total monocytes counts.
Fig. 2C shows total immature cells counts.
Abbreviations: cont (control, untreated), veh (vehicle), ce (cell), T (time — days)
Fig. 3 - OGP(10-14) stimulates the number of circulating double positive
CD34t/Sca-1+ in mice undergoing chemoablation of hematopoietic tissues
Male ICR mice weighing 25 gm each were subjected to chemoablation using cyclophosphamide (CFA), 5 mg/mouse, injected intraperitoneally on days 0 and 1, one injection each day. OGP(10-14) was dissolved in “sterile water for injection” at 100 nmol/ml concentration and 0.1 ml of this solution or water only (vehicle) was administered subcutaneously in the nape daily from day -7 to day —1 and from day +2 to day +8. CFA ablated mice treated with 108 UIl/0.1 ml G-CSF from day +2 to +8 served as positive reference. Data are mean + SD (error bars were too small to be displayed) obtained in 33 animals per condition.
Abbreviations: veh (vehicle), T (time — days), *: significant over
CFA-+vehicle, p<0.01.
Figs. 4A-C - Effect of OGP(10-14) treatment regimen on ex vivo colony forming units derived from bone marrow of mice subjected to chemoablation of hematopoietic tissues
Male ICR mice weighing 25 gm each were subjected to chemoablation using cyclophosphamide (CFA), 5 mg/mouse, injected intraperitoneally on days 0 and 1, one injection each day. OGP(10-14) was dissolved in “sterile water for injection” at 100 nmol/ml concentration and 0.1 ml of this solution or water only (vehicle) was administered subcutaneously in the nape daily for the indicated period(s). Bone marrow was harvested on day 9 and analyzed for colony forming units. Data are meam=SD obtained in animals per condition.
Fig. 4A shows CFU-GM.
Fig. 4B shows CFU-GEMM.
Fig. 4C shows BFU-E.
Abbreviations: Colo/di (colonies/dish), Veh (vehicle).
Figs. 5A-B ~ Microphotography of bone marrow biopsy
Fig. BA presents photomicrographs of two parts of bone marrow specimen from idiopatic myelofibrosis (IMF) patient cultured ex vivo for : 14 days in the absence of OGP (10-14).
Fig. 5B presents photomicrographs of two parts of bone marrow specimen from idiopatic myelofibrosis (IMF) patient cultured ex vivo for 14 days in the presence of 10-8 M OGP (10-14). Note increased cell density in specimen cultured with OGP(10-14).
Figs. 6A-B - Microphotography of bone marrow biopsy
Fig. 6A presents photomicrographs of reticulum stained sections from two parts of bone marrow specimen from idiopatic myelofibrosis (IMF) patient cultured ex vino for 14 days in the absence of OGP(10-14).
Fig. 6B presents photomicrographs of reticulum stained sections from two parts of bone marrow specimen from idiopatic myelofibrosis (IMF) patient cultured ex vivo for 14 days in the presence of 10-8 M OGP(10-14).
Note normal appearance of OGP(10-14) treated tissue.
Fig. 7 - Regression analysis in IMF
Regression analysis in idiopatic myelofibrosis (IMF) patients between hemoglobin level and ex vivo ratio of hematopoietic cells number in
OGP(10-14) treated over untreated specimens (T/C ratio), suggesting a direct relationship between the severity of IMF and effect of OGP(10-14).
Abbreviations: Hem(hemoglobin), Hemato(hematopoietic), rat(ratio), cellu (cellularity).
A number of methods of the arts of cell biology and peptide chemistry are not detailed herein, being well known to the person of skill in the art.
Such methods include peptide synthesis and structural analysis,
differential cell counts, cell sorting analyses, colony forming assays, and the like. Textbooks describing such methods are e.g., Current Protocols in
Immunology, Coligan et al. (eds), John Wiley & Sons. Inc., New York, NY ) and Stewart, JM. and Young J.D., In: Solid Phase Peptide Synthesis,
Pierce Chemical Co., Rockford, IL, pp. 1-175 (1984). These publications are incorporated herein in their entirety by reference. Furthermore, a number of immunological techniques are not in each instance described herein in detail, as they are well known to the person of skill in the art.
The following abbreviations are used herein:
OGP(s) - osteogenic growth polypeptide(s).
OGPBP(s) - osteogenic growth polypeptide binding protein(s). sOGP - Synthetic OGP.
WBC - (white blood cells). ~ PBL - (peripheral blood).
CFA- (cyclophosphamide).
BMT - (bone marrow transplantation).
IMF — (idiopatic myelofibrosis).
Several cellular or soluble agents could be responsible for the interaction between bone and bone marrow cells. This interaction seems to be essential for the regulation of commitment, proliferation, and differentiation of hematopoietic stem and progenitor cells.
OGP increases osteogenesis and bone marrow cellularity [Greenberg, Z., et al., ibid. (1993); Gurevitch, O., et al., ibid. (1996)]. Moreover, 0GP is a potent mitogen for osteoblastic and fibroblastic cells and bone marrow stromal cells [Greenberg, Z., et al., J. Cellular Biochem, 65:359-367 (1997);
Robinson, D., et al, J. Bone Min. Res., 10:690-696 (1995)].
In an osteoblastic cell line, it has recently been reported that OGP activates mitogen-activated protein kinase via a pertussis toxin-sensitive
G-protein. These activities appear to be restricted to C-terminal pentapeptide OGP(10-14) and, therefore, it has been suggested that
OGP(10-14) is the bioactive form of OGP [Bab, I., et al., ibid. (1999).
OGP(10-14) could be extremely interesting in view of a possible in vivo utilization, considering the absence of immunogenicity and toxicity and the relative simplicity of the production and handling of the peptide.
Previous studies have demonstrated that after daily s.c. injections of 0.1 to 10 nmol OGP for 2 weeks in normal mice, the peptide induced an increase greater than 50% in the WBC counts and approximately 40% : enhancement of overall bone marrow cellularity [Gurevitch, O., et al, ibid., (1996)]. The proportion of different cell types was not altered by the treatment, which suggests a multilineage activity on hematopoiesis.
Interestingly, in the experiment doscribed herein, after revereible aplasia induced by the administration of CFA (cyclophosphamide), mice treated by OGP(10-14) recovered faster than those injected with placebo and without any valuable toxicity at the employed doses.
Thus, in a first aspect the present invention relates to a pharmaceutical composition comprising as an effective ingredient at least one oligopeptide having stimulatory activity on the production of hematopoietic cells, preferably having the amino acid sequences
Tyr-Gly-Phe-Gly-Gly, Tyr-Gly-Phe-His-Gly, Gly-Phe-Gly-Gly or
Met-Tyr-Gly-Phe-Gly-Gly, also denoted by SEQ ID NOs:1, 2, 3, and 4, respectively, and a pharmaceutically acceptable carrier.
The process of blood cell formation whereby red and white blood cells are replaced through the division of cells located in the bone marrow is called ‘ hematopoiesis. For review of hematopoiesis see Dexter and Spooncer [Ann. Rev. Cell Biol., 3:423-441 (1987).
There are many different types of blood cells, which belong to distinct cell lineages. Along each lineage, there are cells at different stages of maturation. Mature blood cells are specialized for different functions. For example, erythrocytes are involved in Oz and CO: transport; T and B lymphocytes are involved in cell and antibody mediated immune responses, respectively; platelets are required for blood clotting; and the granuloctyes and macrophages act as general scavengers and accessory cells. Granulocytes can be further divided into basophils, eosinophils, neutrophils and mast cells.
In a specifically preferred embodiment of the present aspect, the pharmaceutical composition of the invention comprises an oligopeptide which is a pentapeptide having the formula: Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NO:1. This pentapeptide is designated OGP(10-14) throughout the present application.
In another embodiment, the pharmaceutical composition of the invention comprises an oligopeptide which is a pentapeptide having the formula:
Tyr-Gly-Phe-His-Gly, as denoted by SEQ ID NO:2.
In yet another embodiment, the pharmaceutical composition of the mvention comprises an oligopeptide which is a tetrapeptide having the formula: Gly-Phe-Gly-Gly, as denoted by SEQ ID NO:3.
In another embodiment, the pharmaceutical composition of the invention comprises an oligopeptide which is a hexapeptide having the formula
Met-Tyr-Gly-Phe-Gly-Gly, as denoted by SEQ ID NO:4, in which the . methionine residue may be acylated.
The peptides used as the effective ingredient in the pharmaceutical compositions of the invention are synthetically produced by known organic chemistry methods. Such synthesis is described, for example, in said US Patent No. 5,814,610.
According to a preferred embodiment of the present aspect, the : pharmaceutical composition of the invention is intended for enhancement of engraftment of bone marrow transplants, hematopoietic reconstruction, bone marrow re-population and the number of circulating hematopoietic stem cells.
According to another embodiment, the pharmaceutical composition of the mvention is intended for enhancement of engraftment of bone marrow transplants, hematopoietic reconstruction, bone marrow re-population and the number of circulating hematopoietic stem cells of a patient receiving chemotherapy or irradiation.
The capacity of the hematopoietic stem cells to provide for the lifelong production of all blood lineages is accomplished by a balance between the stem cell plasticity, that is the production of committed progenitors cells which generate specific blood lineages, and the stem cell replication in the undifferentiated state (self-renewal). The mechanism regulating hematopoietic stem cell plasticity and self-renewal in vivo have been difficult to define. However, the major contributory factors represent a combination of cell intrinsic and environmental influences [Morrison, et al., Proc. Natl. Acad. Sci. USA 92:10302-10306 (1995)]. The importance of the hematopoietic microenvironment has been established through the use of long-term bone marrow culture systems where hematopoietic cells cultured on stroma allow for the maintenance of HSCs, albeit at low frequencies [Fraser, et al, Proc. Natl. Acad. Sci. USA 89 (1992); :
Wineman, et al., Blood 81:365-372 (1993)].
The demonstration of hematopoietic cell maintenance in culture has led to efforts to identify candidate ‘stem cell’ factors. The role of hematopoietic cytokines in stem cell maintenance has been studied by direct addition of purified factors to in vitro cultures of stem cell populations followed by transplantation of the cultured cells [Meunch, et al., Blood 81:3463-3473 (1993); Wineman et al., ibid. (1993); Rebel, et al.,
Blood 83:128-136 (1994)]. Most of the known ‘early-acting’ cytokines such as IL-3, IL-6 and KL have been shown to stimulate proliferation of more committed progenitor cells while concurrently allowing maintenance but not expansion, of cells capable of long-term multilineage repopulation [reviewed in Williams, Blood 81(12):3169-3172 (1993); Muller-Sieburg and Deryugina, Stem Cells; 13:477-486 (1995)]. While these data indicate that the cells plasticity and repopulating function may be preserved by cytokine treatment, the molecules that promote self-renewal of these pluripotent cells remain unknown.
The polypeptide used in the pharmaceutical composition of the invention has been shown to increase the percentage of circulating multilineage progenitor cells. These multilineage progenitor cells are the circulating early precursor CD34 positive cells.
In the human and mouse, primitive mature hematopoietic progenitor cells can be identified a belonging to a class of cells defined by their expression of a cell surface antigen designated CD34. These cells may be referred to as CD34 positive cells. In the mouse, an early subclass of the
CD34 positive hematopiotic cells are the double positive CD34* /Sca* cells.
The analogous Sca-1 cell surface antigen in the human is Flk2. Therefore, human CD34/Flk2 double positive cells are considered equivalent to the mouse double positive CD34/Sca-1 cells.
Human hematopoietic progenitor cells which express the CD34 antigen and/or the Flk 2 receptor are referred to herein as “primitive progenitor cells.” By contrast, hematopiotic cells which do not express either the
CD34 antigen or the flk2 receptor are referred to as “mature progenitor cells.” Therefore, as preferred embodiment the multilineage progenitor cells are the circulating early precursor CD34/Flk2 double positive cells.
As used herein, “progenitor cell” refers to any somatic cell, which has the capacity to generate fully differentiated, functional progeny by differentiation and proliferation. Progenitor cells include progenitors from any tissue or organ system, including, but not limited to, blood, nerve, muscle, skin, gut, bone, kidney, liver, pancreas, thymus, and the like. Progenitor cells are distinguished from “differentiated cells’, which are defined as those cells which may or may not have the capacity to proliferate, i.e., self-replicate, but which are unable to undergo further differentiation to a different cell type under normal physiological conditions. Moreover, progenitor cells are further distinguished from abuorimal cells such as cancer cclle, cepecielly leukemia cells, which proliferate (self-replicate) but which generally do mot further differentiate, despite appearing to be immature or undifferentiated.
Progenitors are defined by their progeny, e.g., granulocyte/macrophage colony-forming progenitor cells (GM-CFU) differentiate into neutrophils or macrophages; primitive erythroid blast-forming units (BFU-E) differentiate into erythroid colony-forming units (CFU-E) which give rise to mature erythrocytes. Similarly, the Meg-CFU, GEMM-CFU, Eos-CFU and Bas-CFU progenitors are able to differentiate into megakaryocytes, granulocytes, macrophage, eosinophls and basophils, respectively.
Various other hematopoitic progenitors have been characterized. For example, hematopoietic progenitor cells include those cells, which are capable of successive cycles of differentiating and proliferating to yield up to eight different mature hematopoietic cells lineages. At the most primitive or undifferentiated end of the hematopictic spectrum, hematopoietic progenitor cells include the hematopietic “stem cells.”
These rare cells, which represent 1 in 10,000 to 1 in 100,000 of cells in the bone marrow, each have the capacity to generate >1013 mature blood cells of all lineages and are responsible for sustaining blood cell production over the life of an organism. They reside in the bone marrow primarily in a quiescent state and may form identical daughter cells through a process called self-renewal. Accordingly, such an uncommitted progenitor can be described as being “omnipotent,” i.e., both necessary and sufficient for generating all types of mature blood cells. Progenitor cells which retain a capacity to generate all blood cell lineages, but which cannot self-renew are termed “pluripotent”. Cells which can produce some but not all blood lineages and cannot self-renew are termed “multipotent.”
The oligopeptides used in the invention are useful in preserving any of these progenitors cells, including unipotent progenitor cells, pluripotent progenitor cells, and/or omnipotent progenitor cells. The oligopeptides, and particularly OGP(10-14), demonstrate particular efficacy in preserving hematopoietic progenitor cells.
In a further preferred embodiment, the oligopeptide used as an effective ingredient in the pharmaceutical composition of the invention enhances the immature cell monocyte recovery and selectively increases any one of the BFU-E and GEMM colony forming units (CFU).
Example 3 below describes ex vivo assessment of hematopoietic colony formation derived from OGP(10-14) and control treated mice. The results indicate an increase of GEMM-CFU and BFU-E in cultures derived from
OGP(10-14)-treated mice compared to the vehicle only control group, whereas a positive G-CSF control induces a significant increase of
GM-CFU. The increases in colony formation in cultures derived from
OGP(10-14) treated mice where apparent only when the treatment began seven days prior to chemoablation. Beth in vive and ex vivo results reached by OGP(10-14) confirm the previously reported multilineage activity of full-length OGP compared to different cytokines. Differently to other growth and mobilizing factors [Fleming, W., et al., Proc. Natl. Acad.
Sci. USA 90:3760 (1993)], sOGP(10-14) increases the number of hematopoeitic stems cells in the peripheral blood without reducing the bone marrow stem cells compartment. :
The pharmaceutical composition of the invention may therefore be intended for increasing the number of white blood cells (WBC), circulating hematopoietic stem cells, and overall bone marrow cellularity.
In a specifically preferred embodiment, the composition of the invention 1s intended for supporting bone marrow transplantation. This effect is due to the activity of the oligopeptides that increases the number of stem cells, accelerates the hematopoietic reconstruction upon bone marrow transplantation and increases the cellularity of bone marrow.
As described in Example 1, the oligopeptides of the invention have been found to enhance the engraftment of bone marrow transplants and to stimulate hematopoietic reconstruction. Bone marrow transplantation (BMT) is progressively and rapidly becoming the treatment of choice in instances of hematological malignancies such as lymphomas, Hodgkin's diseases and acute leukemia as well as solid cancers, in particular melanoma and breast cancer. Recently, BMT is increasingly being taken into consideration in treatment of myeloproliferative disorders such as
IMF (idiopathic myelofibrosis). Potentially, with improved methods, BMT can also be used for treating other catastrophic diseases - AIDS, aplastic anemia and autiommune disorders. The aim of all BMT is to replace the host hematopoietic stem cells, omnipotent and pluripotent, injured by chemotherapy, radiation or disease. These stem cells can replicate : repeatedly and differentiate to give rise to the whole variety of cells present in blood namely erythrocytes, platelets and WBC which include lymphocytes, monoccytes and neutrophils. Resident macrophages and osteoclasts are also derived from hemopoietic omnipotent stem cells. As the stem cells differentiate, they commit themselves more and more to a particular lineage until they can form only one kind of the above cells.
Therefore, according to another specifically preferred embodiment, the pharmaceutical composition of the invention may be used in treating bone marrow transplanted subjects suffering from a hematological disorder, solid tumor, immunological disorder or aplastic anemia. More specifically, the hematological disorder may be a lymphoma, Hodgkin's disease or acute leukemia and myeloproliferative disorder, particularly idiopathic myelofibrosis (IMF).
In IMF bone erythropoiesis evolves to a progressive failure, whereas ectopic hemopoiesis develops and increases. Pathological calcification of fibrosis and structural alterations of trabecular bone may be responsible for an absolute or relative deficit of osteoblasts secreted factors and thus, at least partially responsible for the impaired bone marrow function.
The results described in Example 4, strongly suggest that the OGP(10-14) can increase the hematopoietic cell density of bone marrow in cultured bone fragments from IMF patients without modifying, in such a short time, the fibrosis. The cell increment appears to be balanced and it does not account for the expansion of atypical cells. One may not exclude, of course, that OGP(10-14) simply preserves in culture the bone marrow structure and cellularity of IMF samples compared to those found in samples cultured without the pentapeptide. However, the preserved or even increased cellularity in some OGP(10-14) cultured samples compared to that found in the native ones, suggests a proliferative activity of the peptide. It is not clear, at present, if OGP acts on blood precursors directly or via stromal cells or different cell populations but, at least at the morphological level, its activity appears independent of a significant remodelling of microenvironment.
One implication of this observation is that OGP(10-14) is, in fact, able to enhance, in vitro, three lineage expansion of human hematopoietic cells.
The pharmaceutical compositions of the invention comprise as active i ingredient an oligopeptide as described above, or a mixture of such oligopeptides, in a pharmaceutically acceptable carrier, excipient or stabilizer, and optionally other therapeutic constituents. Acceptable carriers, excipients or stabilizers are non-toxic to recipients at the dosages and concentrations employed, and include buffers, such as phosphate buffered saline and like physiologically acceptable buffers, and more generally all suitable carriers, excipients and stabilizers known in the art, e.g., for the purposes of adding flavors, colors, lubrication, or the like to the pharmaceutical composition.
Carriers may include starch and derivatives thereof, cellulose and derivatives thereof, e.g., microcrystalline cellulose, Xantham gum, and the like. Lubricants may include hydrogenated castor oil and the like.
A preferred buffering agent is phosphate-buffered saline solution (PBS), which solution is also adjusted for osmolarity.
A preferred pharmaceutical formulation is one lacking a carrier. Such formulations are preferably used for administration by injection, including intravenous injection.
The preparation of pharmaceutical compositions is well known in the art and has been described in many articles and textbooks, see e.g.
Remington’s Pharmaceutical Sciences, Gennaro A. R. ed.,, Mack :
Publishing Company, Easton, Pennsylvania, 1990, and especially pages 1521-1712 therein.
The pharmaceutical compositions of the invention can be prepared in dosage units forms. The dosage forms may also include sustained release devices. The compositions may be prepared by any of the methods well known in the art of pharmacy. Such dosage forms encompass physiologically acceptable carriers that are inherently non-toxic and non-therapeutic. Examples of such carriers include ion exchangers, alumina, aluminum stearate, lectithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts, or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose- based substances, and PEG. Carriers for topical or gel-based forms of these polypeptides include polysaccharides such as sodium carboxymethylcellulose or methylcelluslose, polyvinylpyrrolidone, polyacrylasts, polyoxyethylene-block polymers, PEG, and wood was alcohols. For all adminstratioins, conventional depot forms are suitably used. Such forms include for example, microcapsules, nano-capsules, liposomes, plasters, inhalation forms, nose sprays, sublingual tablets, and sustained-release preparations.
Suitable examples of sustained-release preparations include semi-permeable matrices of solid hydrophobic polymers containing the oligopeptides according to the invention, which matrices are in the form of shaped articles, e.g. films, or micro-capsules. Examples of sustained-release matrices include polyesters, hydrogels, polylactides as described by, (U.S. Pat. No. 3,377,919), copolymers of L-glumatic acid and : y—ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the Lupron Depots™ (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acitate), and poly-D-(-)3-hydroxybutyic acid. While polymers such as ethylenevinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods. When encapsulated, the peptides remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture of 37°C, resulting in a loss of biological activity and ’ possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggrergation mechanism is discovered to be intermolecular S—S bond formation through- thio-disulfide interchange, stabilization may be achieved by modifying sulfhydryl residues, lyophilizing from acidic ‘solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.
Sustained-release oligopeptides and particularly the sOGP1-14 compositions alee include lincomally entranned polypeptides, Lincomes containing these polypeptides are prepared by methods known in the art, such as described in Eppstein, et al., Proc. Natl. Acad. Sci. USA 82:3688-3692 (1985); Hwang, et al., Proc. Natl. Acad. Sci. USA 77:4030 (1980); US Patents Nos. 4,485,045 and 4,544,545. Ordinarily, the lipsomes are the small (about 200-800 Angstroms) unilamelar type in which the lipid content is greater than about 30 mol.% cholesterol, the selected proportion being adjusted for the optimal polypeptides therapy.
Lipsomes with enhanced circulation time are disclosed in US Patent No. 5,013,556.
Therapeutic formulations of the oligopeptides are prepared for storage by mixing these polypeptide having the desired degree of purity with optional physiologically acceptable carriers, excipients, or stabilizers [Remington’s Pharmaceutical Sciences, 16h edition, Osol, A., Ed., (1980)], : in the form of lyophilized cake or aqueous solutions. Acceptable carriers, excipients, or stabilizers are non-toxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbhydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol and sorbitol; slat-forming counter-ions such a sodium; and/or non-ionic surfactants such as Tween, Pluronics™ or polyethlene glycol (PEG).
The oligopeptides may also be entrapped in micro-capsules prepared, for example, by coacervation techinques or by interfacial polymerization (for example, hydroxymethylcellulose or gelatine-microcapsules and poly-(methylmethacylate) microcapsules, respectively), in colloidal drug delivery system (for example, liposomes, albumin microshperes, microemulsions, nanoparticles, and nanocapsules), or in macroemulsions.
Such techniques are disclosed in Remington’s Pharmaceutical Sciences, ibid.
The pharmaceutical composition is preferably for a once daily use by a subject in need, and preferably comprises a dosage of active ingredient of about 0.001 to about 50 nmol, more preferably about 0.05 to 25 nmol, most preferably about 0.1 to about 10 nmol.
It is to be appreciated that in addition to the described oligopeptides, the transplantation-supporting composition of the present invention may further optionally comprise other therapeutic constituents. Such constituents may be one or more known cytokines, for example, IL-3,
IL-4, IL-5, G-CSF, GM-CSF (granulocytemacrophage colony stimulating factor) and M-CSF (macrophage colony stimulating factor). When such further component is incorporated in the composition, the effect of the composition in supporting bone-marrow transplantation can increase synergistically.
As a second aspect, the present invention relates to the use of any of the above described oligopeptides, particularly Tyr-Gly-Phe-Gly-Gly,
Tyr-Gly-Phe-His-Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly, as denoted by SEQ ID NOs:1, 2, 8, and 4, respectively, in the preparation of a pharmaceutical composition for enhancement of engraftment of bone marrow transplant, hematopoietic reconstruction, bone marrow re-population and number of circulating stem cells.
In addition, the oligopeptides described herein may be used in the preparation of pharmaceutical compositions for accelerating the engraftment of bone marrow transplants, enhancing proliferation of transplanted stem cells and thus increasing the availability of all types of hematopoietic cells including erythrocytes and thus obviating the need tor supporting the host with these ceils for ai ieasi several weeks; enhancing stromal hematopoietic microenvironment by increasing the stromal cells number and/or expression of stromal cell derived factors that support hemopoiesis; enhancing the hematopoietic stem cell expression of receptors to factors that support hemopoiesis; enhancing the "homing” of intravenously administered bone marrow transplants to the host bone marrow; enhancing the restoration of blood cellularity after
BMT; enabling successful transplantation using reduced cell number, thus decreasing the number of (multiple) marrow extractions from donors and enabling the use of transplants as small as 10-15 ml (instead of 1000 ml); increasing the number of hematopoietic omnipotent and/or pluripotent stem cells in the donor peripheral blood, thus improving the feasibility of transplanting stem cells from peripheral blood; increasing the number of hematopoietic stem cells in vitro in long-term bone marrow cultures for use as transplants and also providing for a method of inhibiting growth of tumor cells in allografts from leukemia patients; enhancing the endogenous restoration of marrow and blood cellularity after chemo- and/or radiotherapy; and enhancing the restoration of population of resident macrophages after BMT or after chemo- and/or radiotherapy.
The magnitude of a therapeutic dose of an oligopeptides or the composition of the invention will of course vary with the group of patients (age, sex, etc.), the nature of the condition to be treated and with the particular oligopeptide employed and its route of administration. In any case the therapeutic dose will be determined by the attending physician.
Any suitable route of administration may be employed for providing a mammal, especially a human, with an effective dosage of a polypeptide of this invention. Intravenous, subcutaneous and oral administration may be preferred.
As a preferred embodiment, these oligopeptides are used for the preparation of a pharmaceutical composition for increasing the circulating multilineage progenitor cells percentage. These multilineage progenitor cells are the circulating early precursor CD34 positive cells, and preferably, CD34/F1k2 double positive cells.
A “hematopoietic stem/progenitor cell” or “primitive hematopoietic cell” as described above, is a cell which is able to differentiate to form a more committed or mature blood cell type. A “hematopoietic stem cell” or “stem cell” 1s one that is specifically capable of long-term engraftment of a lethally irradiated host.
A “CD34* cell population” is enriched for hemotopoietic stem cells. A
CD34% cell population can be obtained from umbilical blood or bone marrow, for example. Human umbilical cord blood CD34* cells can be selected for using immunomagnetic beads sold by Miltenyi (Calfornia), following the Manufacturer's directions.
Furthermore, the oligopeptides used for the preparation of the pharmaceutical composition of the invention enhance the immature cell and monocyte recovery and selectively increases any one of the BFU-E and GEMM colony forming units (CFU). :
Accordingly, such oligopeptides are used in the preparation of the pharmaceutical composition for increasing the number of white blood cells (WBC), circulating hematopoietic stem, and overall bone marrow cellularity.
More specifically, the invention provides the use of these polypeptides in the preparation of a pharmaceutical composition for supporting bone marrow transplantation. This effect is due to the activity of the oligopeptides in increasing the numhar of ctem cella accelerating the hematological reconstruction upon bone marrow transplantation and increasing the cellularity of bone marrow.
According to another specifically preferred embodiment, the present invention relates to the use of said oligopeptides in the preparation of a pharmaceutical composition for treating a subject suffering from hematological disorders, solid tumors, immunological disorders and aplastic anemia. More specifically, the hematological disorder may be a lymphoma, leukemias, Hodgkin's disease and myeloproliferative disorders, particularly idiopathic myelofibrosis (IMF).
In a third aspect, the present invention provides a method for enhancement of engraftment of bone marrow transplant, hematopoietic reconstruction, bone marrow re-population and number of circulating : stem cells. This method comprises administering to a subject in need thereof, an effective amount of an oligopeptide having stimulatory activity on hematopoietic cells as described above, or of a composition of the invention.
According to another embodiment, the invention provids a method for enhancement of engraftment of bone marrow transplant, hematopoietic reconstruction, bone marrow re-population and number of circulating stem cells in patients receiving chemotherapy or irradiation.
In yet another embodiment, an effective amount of the oligopeptides or the composition of the invention may be used to improve engraftment in bone marrow transplantation or to stimulate mobilization and/or expansion of hematopoietic stem cells in a mammal prior to harvesting hematopoietic progenitors from the peripheral blood thereof.
According to a specific embodiment of this aspect, the invention relates to a method of treating a subject suffering from a hematological disorder, solid tumor, immunological disorder or aplastic anemia, by administering to the subject a therapeutically effective amount of an oligopeptide having stimulatory activity on production of hematopoietic cells, or of a composition comprising the same according to the invention.
In another specific embodiment, this method can be used in support of the treatment of the subject by bone marrow transplantation.
For therapeutic applications, the oligopeptides or the pharmaceutical composition useful according to the invention are administered to a mammal, preferable a human, in a physiologically acceptable dosage from, including those that may be administered to a human intravenously as a bolus or by continuous infusion over a period of time.
Alternative routes of administration include intramuscular, intraperitoneal, intra-cerebrospinal, subcutaneous, intra-articular, intrasynovial, intrathecal, oral or topical routes. The oligopeptides or the compositions of the invention also are suitably administered by intratumoral, peritumoral, intralesional, or perilesional routes or to the lymph, to exert local as well as systemic therapeutic effects.
The oligopeptides or the pharmaceutical compositions to be used for in vivo administration must be sterile, This is readily accomplished by filtration through sterile filtration membranes, prior to or following lypophillization and reconstitution. Oligopeptides may be stored in solution. Therapeutic oligopeptides compositions generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.
An “effective amount” of any of the oligopeptides or compositions of the invention to be employed therapeutically will denend for example ninan the therapeutic objectives, the route of administration, and the condition of the patient. Accordingly, it will be necessary for the therapist to titer the dosage and modify the route of administration as required to obtain the optimal therapeutic effect. Typically, the clinician will administer the oligopeptide until a dosage is reached that achieves the desired effect. A typical daily dosage for systemic treatment might range from about 0.001 nmol/Kg to up to 50 nmolVKg or more, depending on the factors mentioned above.
Another specific embodiment relates to the treatment of a subject carrying a transplant, where an ex vivo method may be adopted. In this method, the cells intended for transplantation are exposed to effective amount of the oligopeptides or compiositions of the invention, prior to their transplantation. ‘
The most common way currently available for acquiring a sufficient amount of hematopoietic stem cells for transplantation is to extract 1 liter or more of marrow tissue from multiple site in the donor’s bones with needle and syringe, an involved process that usually requires general anaesthesia. The donors of allogeneic BMT are usually siblings whose tissue types are compatible and sometimes unrelated donors who are matched to the recipient by HLA typing. Autologous transplants, that eliminate the need for HLA matching may be used in patients undergoing ablative chemoradiotherapy for the eradication of solid tumors.
Autologous stem cells may also be obtained from the umbilical cord blood at birth and stored for future administration.
After transplantation and prior to the establishment of a donor-derived functioning marrow the patients hosting BMT present with a transient marked pancytopenia that exposes them to infections. The incidence of bacterial and fungal infections correlates with both the severity and duration of pancytopenia [Slavin, S. and Nagler, A., Transplantation (1992)]. For a similar reason, the CSF fail to support erythropoiesis and platelet formation.
Oligopeptides that support hematopoiesis may prove useful in other ways as well. Some investigators have found that adding stem cells from the peripheral blood to those from the bone marrow significantly increases the rate of engraftment extracting sufficient numbers of stem cells from peripheral blood is a complicated procedure. Administering such oligopeptides to donors to increase the number of stem cells in the blood will improve the feasibility of transplanting stem cells from peripheral blood [Golde, D.W., Sci. Am. 36 December (1991)].
The prerequisite for hematopoiesis and therefore successful MBT is the
X presence of functional stromal cells and tissue that compromise the hematopoietic microenvironment, determine the homing of the injected stem cells from the circulation to the bone marrow and support hematopoiesis [Watson, J.D. and McKenna, H. J. Int. J. Cell Cloning 10:144 (1992)]. Bone marrow derived stromal tissue also provide the conditions to sustain stem cells in in vitro long-term bone marrow cultures. At present this technology suffices to keep stem cells alive.
Adding the appropriate hemopoietic oligopeptides to these cultures may help expand the stem cell population in vitro, this providing increased numbers of these cells for transplantation.
A combined in vitro/in-vivo approach may provide the basis for a forward-looking strategy for (i) obtaining small stem cell preparations from donor blood or marrow and (ii) healthy individuals to have their stem cells stored for a time when the cells might be needed to treat a serious disease, thus bypassing the complexity associated with the use of allogeneic BMT.
It would therefore be Of Uherapeuiic bupuriance LU use small pepiides such as the oligopeptides described in the present application, that stimulate post- BMT hematopoietic reconstruction by enchancing in vivo, ex vivo and/or in vitro the hematopoietic microenvironment of which fibrous tissue, bone and bone cells are important components. Such peptides may also support hematopoiesis in spontaneously occurring or induced myelosuppression condition that do not necessarily involved
BMT.
The oligopeptides described in the present application and preferably the pentapeptide OGP(10-14), appear to directly act at the level of the early hematopoietic precursor (i.e., hematopoietic stem/progenitor cells). Such an expanded stem cell population can serve as the source of cells for myelopoiesis, erythropoiesis (e.g, splenic erythropoiesis) and lymphopoiesis. Accordingly, these oligopeptides can be used to stimulate } proliferation and/or maintenance of hematopoietic stem/progenitor cells either in vitro or in vive (e.g. for treating hematopoietic diseases or disorders).
Therefore, a preferred embodiment relates to a method for enhancing the proliferation of hematopoietic stem/progenitor cells. According to the invention, this method comprises the steps of exposing these cells to an effective amount of an oligopeptide having stimulatory activity on hematopoietic cells, or to an effective amount of a composition comprising the same, as described above. According to the invention such exposure is effective in enhancing the proliferation of said cells.
The term “enhancing proliferation of a cell” encompasses the step of increasing the extent of growth an/or reproduction of the cell relative to an untreated cell either in vitro or in vive. An increase in cell proliferation in cell culture can be detected by counting the number of cells before and after exposure to a molecule of interest. The extent of proliferation can be quantified via microscopic examination of the degree of confluency. Cell proliferation can also be quantified using a thymidine or BrdU incorporation assay.
In a specifically preferred embodiment, the method of the invention is intended for enhancing the proliferation of a CD34 positive cells, preferably F1k2 positive cells.
The oligopeptides or the compositions of the invention are useful in in vivo or ex vivo enhancing the number and/or proliferation and/or differentiation and/or maintenance of hematopietic stem/progenitor cells, expand population of these cells and enhance repopulation of such cells and blood cells of multiple lineages in a mammal.
In one specifically preferred embodiment, these cells are in cell culture and therefore, this would be an ex-vivo/in vitro method.
Alternatively, the method of the invention may be used as an in vivo method of treatment, in case that the treated cells are present in a mammal. “Treatment” refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those already with the disease or disorder as well as those in which the disease or disorder is to be prevented. "Mammal” for purposes of treatment refers to any animal classified as a mammal including, human, domestic and farm animals, and Z00, sports, or pet animals, such as dogs, horses, cats, cows, etc. Preferably, the mammal is human.
In a specific embodiment, the mammal treated by the method of the invention is suffering from, or is susceptible to, decreased blood cell levels, which may be caused by chemotherapy, irradiation therapy, bone marrow transplantation therapy or any other iatrogenic or natural cause.
Chemo- and radiation therapies cause dramatic reductions in blood cell population in cancer patients. At least 500,000 cancer patients undergo chemotherapy and radiation therapy in the US and Europe each year and another 200,000 in Japan. Bone marrow transplantation therapy of value in aplastic anemia, primary immunodeficiency, acute leukemia and solid tumors (following total body irradiation) is becoming more widely practiced by the medical community. At least 15,000 Americans have bone marrow transplants each year. Other diseases can cause a reduction in entire or selected blood cell lineages. Examples of these conditions include anemia (including macrocytic and aplactic anemia); thrombocytopenia; hypoplasia; immune (autoimmune) thrombocytopenic purpur (ITP); and HIV induced ITP.
Pharmaceutical products are needed which are able to enhance reconstitution of blood cell populations of these patients.
Accordingly, it is an object of the present invention to provide a method for enhancing the proliferation and/or differentiation and/or maintenance of primitive hematopoietic cells. Such a method may be useful for enhancing repopulation of hematopoietic stem cells and thus mature blood cell lineages. This is desirable where a mammal has suffered a decrease in hematopoietic or mature blood cells as a consequence of disease, radiation or chemotherapy. This method is also useful for generating expanded populations of such stem cells and mature blood cell lineages from such hematopoietic cells ex vivo.
In yet another preferred embodiment, the invention relates to a method for in vitro/ex-vivo maintaining and/or expanding stem cells. This method comprising isolating peripheral blood cells from a blood sample, enriching blood progenitor cells expressing the CD34 antigen, cluttering the enriched blood progenitor cells under suitable conditions, and treating said cells with an oligopeptide having stimulatory activity on hematopoietic cells, or with a composition comprising as an effective ingredient an oligopeptide having stimulatory activity on hematopoietic cells, according to the invention.
In a specific embodiment, the method of the invention might include a further step of exposing the treated cells to a cytokine. As a non-limiting example, such cytokine may be selected from the group consisting of TPO (Thrombopoietin), EPO (Erythropoietin), M-CSF (Macrophage-colony stimulating factor), GM-CSF (Granulocyte-macrophage-CSF), G-CSF (Granulocyte CSF), IL-1 (Interleukin-1), IL-2, IL-3, IL-4, IL-5, IL-6, IL-7,
IL-8, IL-9, IL-10, I-12, LIF (Leukemia inhibitory factor) and XL (Kit ligand).
As an embodiment to an in vivo treatment, the invention relates to ga method for re-populating blood cells in a mammal This method comprises the steps of administering to said mammal a therapeutically effective amount of an oligopeptide having stimulatory activity on ’ hematopoietic cells, or of an effective amount of the composition of the invention. These hematopoietic cells may be any one of erythroid, myeloid and lymphoid cells. : “Lymphoid blood cell lineages” are those hematopoietic precursor cells which can differentiate to form lymphocytes (B-cells or T-cells). Likewise, “lymphopoiesis” is the formation of lymphocytes. "Erythroid blood cell lineages” are those hematopoietic precursor cells which eandifferentiate to form ervthrocvtes (red blood cells) and “erythropoiesis” is the formation of erythrocytes.
The phrase “myeloid blood cell lineages”, for the purposes herein, encompasses all hematopoietic precursor cells, other than lymphoid and erythroid blood cell lineages as defined above, and “myelopoiesis” involves the formation of blood cells (other than lymphocytes and erythrocytes).
Disclosed and described, it is to be understood that this invention is not limited to the particular examples, process steps, and materials disclosed herein as such process steps and materials may vary somewhat. It is also to be understood that the terminology used herein is used for the purpose of describing particular embodiments only and not intended to be limiting since the scope of the present invention will be limited only by the appended claims and equivalents thereof.
It must be noted that, as used in this specification and the appended claims, the singular forms “a”, “an” and “the” include plural referents unless the content clearly dictates otherwise. :
Throughout this specification and the claims which follow, unless the context requires otherwise, the word “comprise”, and variations such as ) “comprises” and “comprising”, will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.
The following examples are representative of techniques employed by the inventors in carrying out aspects of the present invention. It should be appreciated that while these techniques are exemplary of preferred embodiments for the practice of the invention, those of skill in the art, in light of the present disclosure, will recognize that numerous modifications can be made without departing from the spirit and intended scope of the invention.
Reagents 1. C-terminal pentapeptide of Osteogenic Growth Peptide(10-14) [sOGP(10-14)]: Tyr-Gly-Phe-Gly-Gly; M.W. 499.7 (SEQ ID NO:1) was supplied by Polypeptides Laboratories Inc. (Torrance, California 90503,
USA Batch No. 9712-006). 2. CFA — cyclophosphamide (CFA, SIGMA, 5 mg/mouse), was used for induction of marrow ablation. 3. Dexter-like medium: McCoy's Medium (Gibco-Life technologies, USA) . with 12.5% fetal bovine serum (FBS, Hyclone, Holland), 12.5% horse serum (HS, Sigma, St Louis, MO), 0.8% essential and 0.4% non essential aminecacids (Gibco-Life technologies, USA), 1% glutamine (Sigma, St
Louis, MO), 0.4% vitamins including choline, folic acid, inositol, nicotinamide, pyridoxal HC], riboflavin, thiamine HCl, D-Ca pantothenate
(Gibco-Life technologies, USA), 1% amphotericine B (Fungizone,
Bristol-Myers Squibb), 1% gentamicine, and 106 M hydrocortisone in presence of recombinant human stem cell factor (50 ng/mL rhSCF,
Calbiochem, USA), recombinant human granulocyte-monocyte colony stimulating factor (rhGM-CSF 10 ng/ml, Sandoz, Switzerland), recombinant human interleukine-3 (rhIL-3 10 ng/mL, Calbiochem, USA) and recombinant human erythropoietin (rhEpo 2 units/ml, Sigma, St
Louis, MO) with or without sOGP(10-14) 108M (Abiogen Pharma SpA
Research Laboratories). 4. E.D.T.A acid buffer (Mielodec, Bio Optica, Milan, Italy).
Animals * ICR male mice were purchased from Charles River's (Italy) and maintained under specific pathogen-free conditions. * CV57 Black female mice were from the animal facility of the
Hebrew University Medical School (Jerusalem, Israel). The mice of either strain weighed 25 g on their arrival at the inventors’ laboratory.
Statistical analysis
Comparisons between groups were made using a Fisher's PLSD, factorial or for repeated measures, analysis of variance (ANOVA). Mann-Whitney test was used, for colony assays.
Example 1
Effect of OGP(10-14) on engraftment of bone marrow transplant
Materials and methods
CV57 Black female mice were used to investigate the possible effect of
OGP(10-14) on engraftment of bone marrow transplats. OGP(10-14) in phosphate buffered saline was administered by daily subcutaneous, 10 ul mjections for 12 days. The daily dose ranged from 0.001 to 10 nmol per mouse. Control mice received phosphate buffered saline only. On day 8 after the onset of OGP(10-14) treatment the mice were subjected to total body X-ray irradiation consisting of a single 900 rad dose using a 6°Co source (Picker C-9, 102.5 rad/min). This was followed immediately by an mtravenous injection of 105 unselected syngeneic bone marow cells. The animals were sacrificed 14 days after the onset of OGP(10-14) treatment, both femurs were dissected out and their ephiphyseal ends removed. The bone marrow was washed out completely into phosphate buffered saline (PBS). A single cell suspension was prepared by drawing the preparation several times through graded syringe needles and the cells were counted in a hemocytometer.
Results
Fig. 1 shows a stimulatory effect of the OGP(10-14) on the number of post-irradiation/post-transplantation total femoral bone marrow cells.
This effect was dose dependent showing, at the three highest doses, statistically significant , 2-fold increase in cell counts over the PBS controls.
Example 2
Evaluation of the OGP(10-14) toxicity
A shown above, the OGP(10-14) has been found to enhance the engraftment of bone marrow transplants. Threfore, prior to further, detailed analysis of the pharmacological activity of said peptide, the possible toxicity of said peptide was next evaluated.
Fifty-five mice were evaluated for possible OGP(10-14)-related toxicity after 15 days of subcutaneous administration, at the dose of 10 nmol/mouse, and results were compared to those obtained in 30 placebo-treated controls. No differences were found concerning survival, 41
Bn behaviour, body weight gain and gross examination. Concerning the ‘haematological parameters, administration of the reported doses of peptide did not induce any significant modifications in the number of white blood cells (WBC), red blood cells (RBC), platelets (PLT) or haemoglobin (Hb) level.
Example 3
OGP(10-14) Stimulates hematopoietic recovery after bone marrow chemoablation
Materials and Methods
In this set of experiments bone marrow ablation was induced by an intraperitoneal injection of cyclophophamide (CFA, SIGMA, 5 mg/mouse in 150 ( sterile PBS) for two ennsecutive days (designated "day 0" and "day 1". This protocol has been demonstarted to induce severe, reversible leucopoena with L.D. <30 [Spangrude, G.J. et al, Science, 241:58 (1988)].
The lowest bone marrow cell counts were recorded six days after the first injection.
To evaluate the effect of OGP(10-14) on WBC differential cell counts and determine the OGP(10-14) "dose of choice" to be used in further experiments, mice were treated daily by subcutaneous injections of 0.1 ml
OGP(10-14)-free vehicle or vehicle containing different OGP(10-14) doses as outlined in Fig. 2. One group of reference baseline controls was left untreated and received neither CFA nor sterile water vehicle with or without OGP(10-14) (Fig. 2). Blood was collected by retroorbital bleeding on days -12, -4, +3, +7, +14, +17, +21 and +24 (Fig. 2C). Differential cell counts were carried out using a Coulter Counter (Sysmex Microcell :
Counter F-800).
To test the effect of OGP(10-14) on double positive CD34+/Sca-1+ cells in the blood comparatively to that of G-CSF, CFA ablated mice were treated with daily with 10 nmol OGP(10-14) from day -7 until day +7 and blood samples obtained on days +5, +7 and +15 subjected to flow cytometry.
G-CSF was administered on days +2 to +8.
For the flow cytometry, groups of three blood samples from the mice were pooled, and mononuclear cells were obtained by gradient centrifugation and were resuspended in PBS at a concentration of 1 x 106/ml. The cells were then incubated in the presence of specific monoclonal antibodies (final dilution 1:10) for 30 min at 4°C. To detect CD34+ cells, purified rat anti-mouse monoclonal antibody (Pharmingen, RAM34) was used as a first layer. After three washings, the cells were resuspended in PBS and incubated with a FITC polyclonal goat anti-rat (Pharmingen) To detect
Sca-1+/CD34+ cells, samples were further washed three times in PBS and incubated with rat anti-mouse Sca-1 (Ly.6A.2) PE from Caltag.
Substituting the primary antibody with an irrelevant immunoglobulin performed a specific control. Data acquisition and analysis were assessed by FAC-Scan(tm) (Becton Dickinson) flowcytometer using Lysis II software (FIG. 3).
To evaluate different dosing regimens of OGP(10-14), chemoablated mice were subjected to daily treatment with OGP(10-14), as outlined in Fig. 4.
The mice were sacrificed on day +15, the femoral bone marrow was flushed and single cell suspensions (prepared as above) were subjected to ex vivo progenitor cell (colony forming) assays. The OGP(10-14) effect on the formation of CFU-GM, CFU-GEMM and BFU-E was compared to that of G-CSF (Fig. 4).
Progenitor cell assays
Bone marrow cells were recovered on day +10 after injection of CFA from all groups. Cells were diluted to 2 x 106/ml in Iscove’s Modified Dulbecco
MediumIMFM) with 2% FBS and added to methylcellulose medium according to the manufacturer's recommendations (MethoCult, StemCell
Technologies Inc., Vancouver, Canada). 2 x 104 cells were plated in each test. Both M3434 (for murine GM-CFU, and GEMM-CFU) and M3334 for (murine BFU-E assay) were used. M3434 was supplemented with recombinant murine interleukine-3 (rmIl-3, 10 ng/ml), recombinant : human interleukine-6 (rhIL-6, 10 ng/ml), recombinant murine stem cell factor (rmSCF, 50 ng/ml) and recombinant human erythropoietin ( rhEpo, 3 U/ml). InM3434 the only factor included was Epo. Duplicate tests for each mouse were blindly examined after 14 days of incubation according to the protocol procedures.
Results
Total and differential WBC counts carried out on day +3 showd a marked decrease in all CFA-treated groups (Fig. 2). On day 7, there was approximately 2-fold recovery of total WBC counts in the vehicle treated chemoablated, which were still considerably lower than the values recorded in the untreated reference mice. On the other hand, the
OGP(10-14) animals showed higher values at all doses tested, with peak : counts measured in mice receiving 10 nmol OGP(10-14)/day. The counts in this group approached closely those noted in the untreated reference group (Fig. 2A). Differential cell counts carried out on day 7 also demonstrated an OGP(10-14) induced, dose dependent increase in monocyte and immature cell counts (Figs. 2B, 2C). Monocyte counts at the maximal dose (10 nmol) were 6-fold higher compared to the normal reference (Fig. 2B); those of immature cells being also significantly higher than the reference (Fig. 2C). The total WBC counts in all animal groups were normal from day 10 and onwards (Fig. 2A). However, the monocyte counts still presented the same trend seen on day 7 with normal levels being reached on day 14 (Fig. 2B). In spite of a decrease in immature cell counts in all but the 0.01 nmol group, the highest values were still obtained in the 10 nmol group. The immature cell counts were normal in : all groups from day 14 and onwards (Fig. 2C).
The amount of double positive CD34+/Sca-1+ cells on day +5 was 5-fold higher in the chemoablated animals treated daily with 10 nmol
OGP(10-14) than in mice treated with the vehicle alone (Fig. 3). The effect of OGP(10-14) was similar to that of G-CSF. Flow cytometry measurements carried out on days +7 and +15 demonstarted comparable numbers of the CDS34+/Sca-1+ cells. However, on day +15, the
OGP(10-14) treated mice showed a significantly higher number than the vehicle and G-CSF treated mice (Fig. 3).
The progenitor cell assays showed that OGP(10-14) significantly stimulates CFU-GEMM and BFU-E, but not CFU-GM. The effect of OGP was apparent only in instances where the onset of treatment preceded chemoablation by 7 days (Fig. 4). The absence of an OGP (10-14) effect on
CFU-GM is consistent with its non-significant effect on blood granulocyte cell counts. G-CSF had an effect only on the CFU-GM (Fig. 4).
Example 4
OGP 10-14 rescues hematopoietic bone marrow cellularity in ex-vivo samples from patients with idiopathic myelofibrosis
Materials and methods
In order to assess the efficacy of OGP(10-14) in humans, it’s hematopoietic activity was studied in ex vivo bone marrow specimens obtain from patients suffering from idiopathic myelofibrosis (IMF).
Five IMF patients, one scleroderma patient and two patients with other myelodisplastic syndromes (MDS) were enrolled in the study after signing an informed consent. Diagnosis of IMF was performed on the basis of standard clinical and hematological methods [Barosi, G., et al.,
Br. J. Haematol. 104:730-737 (1999)]. Bone marrow biopsy showing fibrosis was an essential feature. The diagnosis of IMF was eventually established after excluding other possible causes of fibrosis and the presence of different myeloproliferative disorders. In particular, the diagnosis of chronic myelogenous leukaemia was ruled out by excluding the presence of Ph chromosome and of ber/abl rearrangement. Three of the five IMF patients had been previously treated by low doses of ) busulfan administered ten days each month and 1 (g 1,25(0H)2D3 day.
The patients’ data are summarized in Table.1.
Table 1: Clinical data of IMF (A-E) and MDS (F-G) patients
Age | Diagn HB | PLT | WBC | LDH |Spleen
Pati | ea | osis |ECO |(g/dl) |®X10% |xy0¢/ | LD | (cm) |Treatme ent | rs) | date G L) L) *¥% int
A 72 10.4 131 15.0 740 17 | Untrea 5/199 | ted
IS
82 2 8.5 434 11.2 830 20 |Treated 9/199 ee
C 78 2 8.2 1.3 664 20 |Untrea 3/200 . ted 0. 70 3 7.3 763 30 |Treated 3/199 0 .
E | 79 1 10.0 180 4.5 18 |Untrea 6/199 ted
F | 65 14.0 24 12.0 408 30 |Treated 4/199 el
G | 65 2 632 11 |Untrea 2/200 ted - 0 .
H | 40 0 15 280 10 300 10 |Untrea
TETITTE
0 * normal values: 240 — 480 U/L + Fcografic measurement
Three mm long bone marrow specimens were obtained from the posterior superior iliac spine by an 8 gauge disposable biopsy needle equipped with a trap to ensure minimal distortion of the specimen (TraoSystem
MDThech, USA). The specimens were divided into three,1 cm long portions. One randomly selected portion was used for preliminary morphological assessment. The remaining two fragments were cultured in 35-mm tissue culture dishes and completely covered by 1 ml of
Dexter-like medium, in the presence of thSCF (50 ng/mL), rhGM-CSF 10 ng/mL), rhil-3 (10 ng/mL), and rhEpo (2 units/mL) with or without 108M
OGP(10-14), at 37°C, 5% COgz-air. Half the medium was changed once, after 7 days, without altering its composition, other than restoring cytokines and OGP(10-14) initial concentration. After additional seven daye in culinre the bone marrow enocimens wore histologically processed.
Briefly, samples were fixed in modified B5, demineralized in ED.T.A acid buffer and sections were stained with Giemsa, hematoxylin-eosin or ) silver impregnation of reticulum. Changes in the bone marrow were assessed semiquantitatively on a I to IV score. A score of IV was used for cell-rich bone marrow specimens comparable to normal ones; score III represented reduced cellularity with reduced nuclear density; score II specimens exhibited spread lacunae; hematopoietic cells in score I specimens were extremely scanty and/or the bone marrow area was vastly substituted by lacunar zones. At least 3 equispaced histological sections per sample were examined using the entire section area. In addition, the cell density was automatically evaluated using a computer assisted Leica microscope equipped with Leica.QWin software, as the ratio between cell counts and bone marrow area. The results for each patient results were expressed as the ratio of mean cell density in
OGP(10-14) treated over untreated specimens (T/C ratio).
Results
After 14 days in culture, bone marrow specimens treated with
OGP(10-14) appear richer in hematopoietic cells than OGP(10-14)-free specimens from the same patients (Figs. 5, 6). The semiquantitative score was significantly increased in all IMF patients (p<0.05). No differences : were detected between OGP(10-14) treated and untraeted specimens from non IMF patients. The computer assisted evaluation of cellularity showed a T/C ratio>1 in all IMF cases (p<0.01) strongly indicating that cell number was increased in the OGP(10-14)-treated specimens.
Moreover, the T/C ratio was statistically significant in every pair of specimens obtained from individual patients (Table. 2). The T/C ratio showed a very high and significant inverse correlation with the patients’ hemoglobin level (Fig. 7). Decreased hemoglobin levels are the most important serological indicator for the severity of IMF. This correlation therefore strongly suggests that the effect of OGP(10-14) is highest in the more severely affected patients.
Table 2: Computer assisted evaluation of cell density.
PATIENT T/C RATIO p<0.05
BB 1 83 1 pool p<0.003 »o - | 81 | p<0.0002 p<0.025
G09 | ~~ p=Ns
The ratio between erythroid and myeloid cells was apparently unchanged after culturing with OGP(10-14). However, a semiquantitative . assessment suggested a 1.5 to 10-fold decrease in the number of megakaryocytes in specimens obtained from IMF patients. As in the case of overall hematopietic cellularity, such differences were not found in samples obtained from the non-IMF patients.
Claims (50)
1. Use of an oligopeptide having a molecular weight of 200 to 1,000 i Da comprising the amino acid sequence of any one of the peptides selected from the group consisting of Tyr-Gly-Phe-Gly-Gly, Tyr- Gly-Phe-His-Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NOs:1, 2, 3 and 4, respectively, in the preparation of a pharmaceutical composition for enhancing the mobilization of multilineage hematopoietic stem cells to the peripheral blood.
2. Use of an oligopeptide having a molecular weight of 200 to 1,000 Da comprising the amino acid sequence of any one of the peptides selected from the group consisting of Tyr-Gly-Phe-Gly-Gly, Tyr- Gly-Phe-His-Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NOs:1, 2, 3 and 4, respectively, in the preparation of a pharmaceutical composition for enhancing the mobilization of multilineage early CD34 positive hematopoietic stem cells to the peripheral blood.
3. The use according to any one of claims 1 and 2, wherein enhancing mobilization to the peripheral blood leads to increase in the number of circulating multilineage early CD34 positive hematopoietic stem cells.
4. Use of an oligopeptide having a molecular weight of 200 to 1,000 Da comprising the amino acid sequence of any one of the peptides selected from the group consisting of Tyr-Gly-Phe-Gly-Gly, Tyr- Gly-Phe-His-Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NOs:1, 2, 3 and 4, respectively, in the preparation of a peripheral blood stem cell transplant for the treatment of a subject in need thereof.
~ 13361/WO/01 ) "
5. The use according to claim 4, wherein said oligopeptide enhances the mobilization of multilineage hematopoietic stem cells to the peripheral blood.
6. The use according to claim 5, wherein said hematopoietic stem cells are CD34 positive cells.
7. Use according to any one of claims 2, 3 and 4, wherein said circulating multilineage stem cells are double positive CD34/F1k2 cells.
8. Use according to any one of claims 1 to 4, wherein said oligopeptide is a pentapeptide having the formula: Tyr-Gly-Phe-Gly-Gly, as denoted by the amino acid sequence of SEQ ID NO: 1.
9. Use according to claims 1 to 4, wherein said oligopeptide is a pentapeptide having the formula: Tyr-Gly-Phe-His-Gly, as denoted by the amino acid sequence of SEQ ID NO:2.
10. Use according to any one of claims 1 to 4, wherein said oligopeptide is a hexapeptide having the formula: Ac-Met-Tyr-Gly-Phe-Gly-Gly, as denoted by the amino acid sequence of SEQ ID NO:4.
11. Use according to any one of claims 1 to 4, wherein said oligopeptide is a tetrapeptide having the formula: Gly-Phe-Gly-Gly, as denoted by the amino acid sequence of SEQ ID NO:3.
12. Use according to any one of claims 1 to 3, in the preparation of a pharmaceutical composition for enhancing the immature cell and monocyte recovery.
0 13361/WO/01 b
13. Use according to any one of claims 1 to 4, in the preparation of a pharmaceutical composition for selectively increasing any one of the BFU-E and GEMM colony forming units (CFU).
14. Use of an oligopeptide having the amino acid sequence of any one of Tyr-Gly-Phe-Gly-Gly, Tyr-Gly-Phe-His-Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NOs:1, 2, 3 and 4 vespectively, in the preparation of a pharmaceutical composition for enhancing the selective proliferation of CD34 positive hematopoietic stem cells in a subject in need thereof.
15. The use according to claim 14, wherein said CD34 positive cells are double positive CD34/F1k2 cells. ©
16. Use of an oligopeptide having the amino acid sequence of any one of Tyr-Gly-Phe-Gly-Gly, Tyr-Gly-Phe-His-Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NOs:1, 2, 3 and 4 respectively, in the preparation of a pharmaceutical composition for treating subjects suffering from a myeloproliferative disorder.
17. Use according to claim 16, wherein said myeloproliferative disorder is idiopathic myelofibrosis (IMF).
18. Use according to any ome of claims 16 and 17, wherein said pharmaceutical composition increases the number of overall bone marrow cellularity in a subject suffering from IMF.
19. Use of an oligopeptide having the amino acid sequence of any one of Tyr-Gly-Phe-Gly-Gly, Tyr-Gly-Phe-His-Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NOs:1, 2,3 : and 4 respectively, for in vitro and/or ex vivo selectively
PCT/IL01/00700 maintaining and/or expanding CD34 positive stem cell population in a blood sample.
20. Use according to claim 19, wherein said blood sample is mammalian blood sample.
21. Use according to claim 20, wherein said blood sample is a human blood sample.
22. Use according to claim 20, wherein said blood sample originates from a mammal suffering from, or susceptible to decreased blood cell counts.
23. Use according to claim 22, wherein said decreased blood counts are caused by chemotherapy, irradiation therapy, or bone marrow transplantation therapy.
24. Use according to claim 23, wherein said composition further comprises at least one cytokine.
25. Use according to claim 24, wherein said cytokine is selected from the group consisting of TPO (Thrombopoietin), EPO (Erythropoietin), M- CSF (Macrophage-colony stimulating factor), GM-CSF (Granulocyte- macrophage-CSF), G-CSF (Granulocyte CSF), IL-1 (Interleukin-1), IL- 2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, LIF (Leukemia inhibitory factor) and KL (Kit ligand).
26. A substance or composition for use in a method for enhancing the mobilization of multilineage hematopoietic stem cells to peripheral blood, said substance or composition comprising an oligopeptide having AMENDED SHEET
PCT/IL01/00700 : the amino acid sequence of any one of Tyr-Gly-Phe-Gly-Gly, Tyr-Gly- Phe-His-Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NOs: 1, 2, 3 and 4 respectively, or of a pharmaceutical composition comprising the same, and said method comprising administering an effective amount of said substance or composition to a subject in need thereof.
27. A substance or composition for use in a method of treatment according to claim 26, wherein said stem cells are multilineage early CD34 positive stem cells.
28. A substance or composition for use in a method for enhancement of the number of circulating multilineage early CD34 positive stem cells, said substance or composition comprising an oligopeptide having the amino acid sequence of any one of Tyr-Gly-Phe-Gly-Gly, Tyr-Gly-Phe-His- Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NOs: 1, 2, 3 and 4 respectively, or of a pharmaceutical composition comprising the same, and said method comprising administering an effective amount of said substance or composition to a subject in need thereof.
29. A substance or composition for use in a method of treatment according to claim 28, wherein said subject in need thereof is a patient receiving irradiation or chemotherapy.
30. A substance or composition for use in a method of treatment according to claim 28, wherein said subject suffers from any one of hematological disorders, solid tumors, immunological disorders and aplastic anemia. AMENDED SHEET
PCT/IL01/00700 ) 31. A substance or composition for use in a method of treatment according to claim 30, wherein said hematological disorder is selected from the group consisting of lymphomas, leukemias, Hodgkin’s diseases and myeloproliferative disorders.
32. A substance or composition for use in a method of treatment according to claim 28, wherein said circulating multilineage stem cells are double positive CD34/F1k2 cells.
33. A substance or composition for use in a method for selectively increasing the number of any one of the BFU-E and GEMM colony forming units (CFU), said substance or composition comprising an oligopeptide having the amino acid sequence of any one of Tyr-Gly- Phe-Gly-Gly, Tyr-Gly-Phe-His-Gly, Gly-Phe-Gly-Gly and Met-Tyr- Gly-Phe-Gly-Gly as denoted by SEQ ID NOs: 1, 2, 3 and 4 respectively, or of a pharmaceutical composition comprising the same, and said method comprising administering an effective amount of said substance or composition to a subject in need thereof.
34. A substance or composition for use in a method of treatment according to claim 33, wherein said subject in need thereof is a patient receiving irradiation or chemotherapy.
35. A substance or composition for use in a method of treatment according to claim 33, wherein said subject suffers from any one of hematological disorders, solid tumors, immunological disorders and aplastic anemia.
36. A substance or composition for use in a method of treatment according to claim 35, wherein said hematological disorder is selected from the AMENDED SHEET
PCT/IL01/00700 ' group consisting of lymphomas, leukemias, Hodgkin's diseases and myeloproliferative disorders.
37. A substance or composition for use in a method for enhancing the number of any one of the BFU-E and GEMM colony forming units (CFU), said substance or composition comprising an oligopeptide comprising the amino acid sequence Tyr-Gly-Phe-Gly-Gly, Tyr-Gly- Phe-His-Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NOs: 1, 2, 3 and 4 respectively, or of a composition comprising said oligopeptide as an effective ingredient, and said method comprising exposing said cells to an effective amount of said substance or composition.
38. A substance or composition for use in a method of treating a subject suffering from a myeloproliferative disorder, said substance or composition comprising an oligopeptide having the amino acid sequence of any one of Tyr-Gly-Phe-Gly-Gly, Tyr-Gly-Phe-His-Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NOs: 1, 2, 3 and 4 respectively or of a composition comprising said oligopeptide as an effective ingredient, and said method comprising administering a therapeutically effective amount of said substance or composition to said subject.
39. A substance or composition for use in a method of treatment according to claim 35, wherein said myeloproliferative disorder is idiopathic myelofibrosis (IMF).
40. A substance or composition for use in a method for enhancing the proliferation of hematopoietic CD34 positive cell stem cells, said substance or composition comprising an oligopeptide having the amino AMENDED SHEET
PCT/IL01/00700 : acid sequence of any one of Tyr-Gly-Phe-Gly-Gly, Tyr-Gly-Phe-His- Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NOs: 1, 2, 3 and 4 respectively, or of a composition comprising said oligopeptide as an effective ingredient, and said method comprising exposing said cells to an effective amount of said substance or composition.
41. A substance or composition for use in a method for selectively increasing the number of hematopoietic CD34 positive cell stem cells, said substance or composition comprising an oligopeptide having the amino acid sequence of any one of Tyr-Gly-Phe-Gly-Gly, Tyr-Gly-Phe- His-Gly, Gly-Phe-Gly-Gly and Met-Tyr- Gly-Phe-Gly-Gly as denoted by SEQ ID NOs: 1, 2, 3 and 4 respectively, or of a pharmaceutical composition comprising the same, and said method comprising administering to a subject in need thereof, an effective amount of said substance or composition.
42. A substance or composition for use in a method of treatment according to any one of claims 40 and 41, wherein said CD34 positive cell is a double positive CD34/Flk2 cell.
43. A substance or composition for use in a method of treatment according to claim 41, wherein said subject is a patient receiving irradiation or chemotherapy.
44. A substance or composition for use in a method of treatment according to claim 41, wherein said subject suffers from any one of hematological disorders, solid tumors, immunological disorders and aplastic anemia. AMENDED SHEET
} PCT/IL01/00700 : 45. A substance or composition for use in a method of treatment and/or prevention according to claim 44, wherein said hematological disorder is selected from the group consisting of lymphomas, leukemias, Hodgkin's diseases and myeloproliferative disorders.
46. A method for in vitro and/or ex vivo maintaining and/or expanding stem cell population in a blood sample comprising isolating peripheral blood cells from said blood sample, enriching blood progenitor cells expressing the CD34 antigen, cluttering the enriched blood progenitor cells under suitable conditions, treating said cells with an oligopeptide having the amino acid sequence of any one of Tyr-Gly-Phe-Gly-Gly, Tyr-Gly-Phe- His-Gly, Gly-Phe-Gly-Gly and Met-Tyr-Gly-Phe-Gly-Gly as denoted by SEQ ID NOs: 1, 2, 3 and 4 respectively or with a composition comprising said oligopeptide as an effective ingredient.
47. Use according to any one of claims 1 to 25, substantially as herein described and illustrated.
48. A substance or composition for use in a method of treatment according to any one of claims 26 to 45, substantially as herein described and illustrated.
49. A method according to claim 46, substantially as herein described and illustrated.
50. A new use of an oligopeptide as defined in any one of claims 1 to 25, a substance or composition for a new use in a method of treatment, or a new In vitro and/or ex vivo method for maintaining and/or expanding stem cell population, substantially as herein described. AMENDED SHEET
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ZA200401552A ZA200401552B (en) | 2004-02-25 | 2004-02-25 | Osteogenic growth oligopeptides as stimulants of hematopoiesis. |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ZA200401552A ZA200401552B (en) | 2004-02-25 | 2004-02-25 | Osteogenic growth oligopeptides as stimulants of hematopoiesis. |
Publications (1)
Publication Number | Publication Date |
---|---|
ZA200401552B true ZA200401552B (en) | 2005-01-26 |
Family
ID=34592875
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
ZA200401552A ZA200401552B (en) | 2004-02-25 | 2004-02-25 | Osteogenic growth oligopeptides as stimulants of hematopoiesis. |
Country Status (1)
Country | Link |
---|---|
ZA (1) | ZA200401552B (en) |
-
2004
- 2004-02-25 ZA ZA200401552A patent/ZA200401552B/en unknown
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP0812201B1 (en) | In vitro amplification of stem cells | |
DeLuca et al. | Prior chemotherapy does not prevent effective mobilisation by G-CSF of peripheral blood progenitor cells | |
US5426098A (en) | Increase in hematopoietic progenitor cells in peripheral blood by transforming growth factor beta | |
CA2735800C (en) | Expansion of haemopoietic precursors | |
CA2109699A1 (en) | Method for improving autologous transplantation | |
EP0831888B1 (en) | Methods for increasing hematopoietic cells | |
WO1998001151A1 (en) | Novel use of mk family as hematopoietic factor | |
EP0752867A1 (en) | Selective cell proliferation | |
US5258367A (en) | Uteroferrin and rose proteins for stimulating hematopoietic cells | |
Piguet et al. | Administration of recombinant interleukin 2 to mice enhances production of hemopoietic and natural killer cells | |
Fazzi et al. | Bone and bone-marrow interactions: haematological activity of osteoblastic growth peptide (OGP)-derived carboxy-terminal pentapeptide. Mobilizing properties on white blood cells and peripheral blood stem cells in mice | |
US20180207202A1 (en) | Methods for mobilizing hematopoietic stem cells | |
RU2360965C1 (en) | METHOD OF INCREASING NUMBER OF PATIENT'S HAEMOPOETIC UNDIFFERENTIATED STEM CELLS ex vivo | |
AU2001282436B2 (en) | Osteogenic growth oligopeptides as stimulants of hematopoiesis | |
US6737051B1 (en) | Cell compositions containing macrophages, presenting anti-infectious and hematopoietic properties | |
AU2001282436A1 (en) | Osteogenic growth oligopeptides as stimulants of hematopoiesis | |
ZA200401552B (en) | Osteogenic growth oligopeptides as stimulants of hematopoiesis. | |
Mahmud et al. | Growth factors mobilize CXCR4 low/negative primitive hematopoietic stem/progenitor cells from the bone marrow of nonhuman primates | |
EP3284473A1 (en) | Agent for inducing amplification of hematopoietic stem cells | |
BG108586A (en) | Osteogenic growth oligopeptides as stimulants of hematopoiesis |