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WO1993015195A1 - Granulines obtenues des leucocytes - Google Patents

Granulines obtenues des leucocytes Download PDF

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Publication number
WO1993015195A1
WO1993015195A1 PCT/CA1992/000089 CA9200089W WO9315195A1 WO 1993015195 A1 WO1993015195 A1 WO 1993015195A1 CA 9200089 W CA9200089 W CA 9200089W WO 9315195 A1 WO9315195 A1 WO 9315195A1
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Prior art keywords
cys
granulin
gly
asp
ala
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PCT/CA1992/000089
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English (en)
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Samuel Solomon
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Samuel Solomon
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Publication of WO1993015195A1 publication Critical patent/WO1993015195A1/fr

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    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475Growth factors; Growth regulators
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides

Definitions

  • This invention relates to novel peptides and pharmaceutical formulations containing them; the pep- tides are useful as inhibitors of keratinocytes.
  • leukocytes are pedtidergic cells.
  • Neutrophil granules contain large amounts of basic, cystine-rich peptides of 29 to 34 amino acids, that have been variously called de- fensins (1), coricostatins (2), myeloid-related sequences (3), and cryptidins (4).
  • de- fensins (1) basic, cystine-rich peptides of 29 to 34 amino acids
  • coricostatins (2) coricostatins (2)
  • myeloid-related sequences (3) myeloid-related sequences
  • cryptidins cryptidins
  • corti- costatins have potential regulatory functions, includ- " ing the ability to inhibit the action of the hormone adrenocorticotropin on glucocorticoid secretion (2,6,7) and to stimulate nifedipine-sensitive L-type Ca ⁇ + channels in villus enterocytes (8). It has also been reported that a human defensin is a monocyte chemotactic agent (9). Other granulocyte-associated peptides have also been shown to have regulatory activities. For example, hemoregulatory peptide 1 is a granulocyte-associated thiol containing pentapep- tide, with potent inhibitory actions on myelopoieses (10).
  • Granulocyte enriched extracts contain several cystine-rich components at levels approximately three orders of magnitude lower than the defensin/corticostatins. From their compositional analysis and chromatographic behaviour these peptides appear unrelated to any known hormone, including the definsin/corticostatins.
  • Gran- ulocyte-derived peptides have potential both as immunoregulatory molecules, and in host resistance.
  • Granulins are novel candidate growth factors recently discovered in human and rat inflammatory leukocytes (22). Two rat granulin homologs, epithelin 1 and 2, occur in the kidney (23). Epithelin 1, which is probably identical to rat leukocyte granulin (22,23) exhibits activities similar to epidermal growth factor on epithelian cells .in vitro (23) .
  • the invention relates to a family of novel leukocyte-associated peptides that are cystine-rich.
  • the peptides are approximately 6 Kda and may be cytokines.
  • Granulin A DVKCDMEVSCPDGYTCCRLQSGAWGC- CPFTQAVCCEDHIHCCPAGFTCDTQKGTCE, SEQ ID NO: 1.
  • Rat Granulin EVKCDLEVSCPDGYTCCRLNTGA G(CCPFSB)AVCCEDHIHCCPAGFTCXTQ, SEQ ID NO: 5.
  • Granulin C VPCDXVSSCPSSDTCCOLTSGEHGCCPIPEAVC, SEQ ID NO: 3.
  • Granulin D IGCDQXDTSSCCPDG, SEQ ID NO: 4.
  • carp Granulin VIHCDAATICPDGTICCLSPYGMBGQCCRDGIHCCRHGYHCDSRTTHC , SEQ ID NO: 6.
  • Granulin B VMCPDARSRCPDGHTCCELPSGKYGCCPMPNATCCSDHLHCCPQDTVCDLIQSK Cl, SEQ ID NO: 2.
  • Granulin E Asp Val Glu Cys Gly Clu Gly His Phe Cys His Asp Asp Gin Thr Cys Cys Arg Asp Asn Arg Glu Gly Trp Ala Cys Cys Pro Try Ala Gin Gly Cla Cys Cys Ala Asp Arg Arg His Cys Cys Pro Ala Gly Phe Arg Cys Ala Arg Arg Gly Thr Lys Cys Leu, SEQ ID NO: 7.
  • Granulin F Ala lie Gin Cys Pro Asp Ser Gin Phe Glu Cys Pro Asp Phe Ser Thr Cys Cys Val Met Val Asp Gly Ser Trp Gly Cys Cys Pro Met Pro Gin Ala Ser Cys Cys Glu Asp Arg Val His Cys Cys Pro His Gly Ala Phe Cys Asp Leu Val His Thr Arg Cys Lie, SEQ ID NO: 8.
  • Granulin G Gly Gly Pro Cys Gin Val Asp Ala His Cys Ser Ala Gly His Ser Cys lie Phe Thr Val Ser Gly Thr Ser Ser Cys Cys Pro Phe Pro Glu Ala Cys Gly Asp Gly His His Cys Cys Pro Arg Gly Phe His Cys Ser Ala Asp Gly Arg Ser Cys Phe, SEQ ID NO: 9.
  • a topical formulation comprising an effective amount of a Granulin of the invention in association with a pharmaceutically acceptable carrier for topical formulation.
  • a method of healing wounds comprising applying to a wound site a Granulin of the invention.
  • FIG. 1 Figure IA shows the HPLC chromatogram. of a crude granule extract from inflammatory exudate cells, and B shows the chromatogram of a whole cell extract. The position of the granulins are marked by arrows. Note the absence in A of thymosin-/3-4, a cyto- plasmic marker peptide.
  • the granule peptide markers HP-1 and HP-4 were identified as previously described (6), lysozyme was identified by a ino terminal sequence analysis (unpublished). Thymosin- ⁇ -4 and its oxidation product were identified by Fast Atom Bombardment mass spectrometry;
  • FIG.2 Size-exclusion purification of granulin A, (panel A), granulin B, (panel B), and granulins C and D, (panel C). Size markers were substance P, CLIP and ACTH1.-39. Apparent m.wts for native granulin A, 2700; granulin B, 3200; granulin C, 1700; and granulin D 7. 3900; FIG. 3 Purification of rat granulin from bone marrow; panel A shows the first HPLC chromatogram in ace onitrile/TFA, and B shows the second step of purification in
  • FIG. 4A Shows the sequence of the probe used to screen a human bone marrow cDNA library in gt 11
  • FIG. 4B Shows a sequencing strategy
  • FIG. 4C Shows the complete nucleotide sequence and deduced polypeptide sequence of granulin
  • FIG. 5 Shows a comparison of the granulin-like domains
  • FIG. 6 Is a Southern blot analysis of digested human DNA
  • FIG. 7A Is a Northern blot analysis of granulin precursor mRNA
  • FIG. 7B Is a Northern blot analysis of RNA from
  • FIG. 7C Shows distribution of granulin mRNA. MODES FOR CARRYING OUT THE INVENTION MATERIALS AND METHODS Tissue Sources
  • S-pyridylethylated peptides were digested using trypsin (TPCK-treated, Sigma) chymotrypsin (Sigma), and S. aureus V8 protease (Sigma), at enzyme to substrate ratios of approximately 1 to 50 by weight. Digestions were performed at 37°C in 100 ⁇ l
  • proteolytic fragments were then fractionated by rp-HPLC on a C-ig ⁇ Bondapak column using a gradient of
  • HPLC profile of a typical extract of human inflammatory cells is shown in Figure 1.
  • HP-1 and HP-4 several low abundance components are present.
  • three low abundance components that had unusually high levels of cystine were identified... These are labelled A, B and C/D, and were present in both whole cell extracts (FIG. IB) and crude granule preparations (FIG. IA) .
  • Each of these extracts was then further purified using size-exclusion HPLC, revealing that the component C/D
  • SUBSTITUTE SHEET contained two peptides, one of which, D, eluted as a larger molecule than the other three. Each peptide was further purified on rp-HPLC. Their amino acid compositions are given in Table 1. The purified peptides were S-pyridylethylated, and amino terminal sequence analyses were performed, revealing that the four peptides were distinct but related molecules with no homology to any known protein. Because these peptides were associated with the granule fraction, we call them granulins A, B, C and D. Granulin D was run on reducing and non-reducing SDS-PAGE, and ran as a smaller molecule after reduction, indicating that it is probably a dimer.
  • Granulin A is the most abundant of the human granulins, and was subject to a more detailed analysis. S-pyridylethylated peptide was digested with trypsin, chy otrypsin or S. aureus V8 protease. The fragments were isolated by HPLC, one fifth aliquots analysed by amino acid analysis, and appropriate fragments were then submitted to gas phase Edman microsequencing. Final recovery of the digestion products was between 150 and 300 picomoles. The overlap of the granulin A fragments is described in the legend to Table 2 together, with similar data for the rat granulin. The two sequences are highly
  • SUBSTITUTE SHEET conserved, as would be expected for regulatory molecules. Inflammatory exudates and bone marrow preparations are mixtures of cells, the exudates containing typically 70 to 95% granulocytes. When leukocytes from the blood of healthy donors was fractionated by density gradient centrifugation, granulins could be detected in the granulocyte pellet, but not in the interface where the mononuclear cells partition (data not shown). The isolation and characterization of a novel family of leukocyte associated cystine-rich peptides, which are called granulins has been described.
  • SUBSTITUTESHEET present it is not certain if both peptides are the product of the same, or different genes. It is also too early to determine whether the epithelins are intrinsic to renal cells, or if they are derived from blood borne cells trapped in the kidney. It is clear, however, that the two peptides from the kidney are members of a larger family, the granulins, and that a major source of the granulins, and probably also the epithelins, is from circulating leukocytes. Rat granulin was isolated from bone marrow, indicating that granulins are of myeloid origin. Whether granulins are also synthesized in circulating leukocytes remains to be determined.
  • Granulins were extracted from granulocyte rich preparations, and are recoverable from the granulocyte pellet after Ficoll- Hypaque density gradient centrifugation. This suggests that their cellular origin may be the neutrophil, however, it is possible that other granulocytes such as eosinophils, or contaminating monocytes, contribute to the granulin content of these extracts. Extracts of human platelets contained no detectable granulins (data not shown). It is possible that each granulin belongs to a distinct cell type, or that sub-classes of the same cell-type contain different granulins. These are issues best answered using immunolocalization procedures. The granulins co-purify in a crude granule extract.
  • Granulocytes have several different granule subclasses, including a true secretory compartment (21) that can be activated independently of phagocytosis.
  • a true secretory compartment (21) that can be activated independently of phagocytosis.
  • suitable immunoassay techniques will permit the unambiguous location .of the granulins to a cell type, and a subcellular compartment.
  • the granulins are a novel family of cystine- rich immunoinflammatory peptides. Their presence in circulating leukocytes and inflammatory exudates, and
  • granulins a novel class of leukocyte peptides with possible cytokine-like activities which are called granulins. They are cystine-rich with molecular weights of approximately 6Kda, except for granulin D, which appears to be a dimer.
  • the sequence of one member of this family, a 56 residue peptide, granulin A, and amino-terminal sequences for three other granulins from human peripheral leukocytes are described.
  • a fifth related peptide was isolated and partially sequenced from rat bone marrow, suggesting that at least some of the granulin in peripheral leukocytes is preformed in the marrow.
  • Rat granulin, and human granulin A are closely related, showing that the granulin structures are highly conserved between species. It has now been found that the precursor for the human granulins is a 593 residue glycoprotein, containing seven repeats of the 12-cys mecanic granulin domain. Gene expression is seen in the kidney, leukemic cell lines and fibroblasts, and very strongly in transformed epithelial cell lines, which both express the gene, and respond to the mature polypeptide.
  • Human leukocytes contain four granulin homologs, designated A, B, C and D. Their cellular distribution, and whether they are products of one or several genes under co-ordinate or independent control was unknown. To address these, and related issues, two oligonucleotide primers for PCR were synthesized corresponding to the amino terminal and mid-portion regions of granulin A (grnA). GrnA was chosen because it is the most abundant granulin in human leukocyte
  • the sequence (Fig. 4C) predicts a protein of 593 residues, with a probable signal peptide (24) extending to residue 17. It contains the 56 residue grnA sequence, as expected, and also six other 12- cysteine granulin-like domains, including B, C and D, hitherto known only from N-terminal sequences.
  • the precursor contains two novel Cys-12 domains, E and F, and a degenerate granulin domain, G, with 10 cysteines.
  • the positions of the cysteines are highly conserved (Fig. 5) as are certain other residues such as ASP39, His42 and Pr ⁇ 4g.
  • An eighth domain at the N- terminus, (paragranulin, Fig. 5) contains only 6 cysteines corresponding to the amino-terminal half of a granulin domain.
  • the inter-domain sequences show little homology, except for a pro-ala dipeptide, found midway between each domain except F and G.
  • Mono- or dibasic sequences which are frequent sites of proteolysis in peptide processing (25), flank granulin domains C, D and E, but not A or B, both of which have been isolated as excised peptides. Thus nothing can be
  • EGF and granulin/epithelins have similar actions on epithelial cells (23), but probably different receptors (23). Whether the similarities are significant, or merely coincidental, is unclear. Unlike the EGF-precursor, pro-granulin has no transmembrane segment or cytoplasmic domain (28,29) implying different post-translational pathways for the two proteins. Multiple sequential repeats of cysteine-rich domains is a recurrent structural motif among regulatory proteins (28,29,30,31), wherein typically, one exon delineates one cysteine-rich domain (30,32,33). In contrast, the PCR amplified fragment of grnA is bisected by an intron (Fig. 4A), and preliminary results show that introns bisect domains G, F, B, C and D.
  • cysteines align with an approximately mirror-image symmetry around the bisecting intron. Genomically, therefore, pro- granulin bears little resemblance to the EGF- precursor.
  • Proerythroid leukemic cells express granulin mRNA (Fig. 7A) , but seem unlikely to be involved in regulating epithelial proliferation. Similarly, the
  • SUBSTITUTE SHEET strong expression of granulin in RNA in kidney, but not other tissues, may imply functions unrelated to epithelial cell mitogenesis.
  • granulins may have multiple biological activities; indeed distinct activities may be associated with different domains, as already proposed for epithelins 1 and 2 (23) .
  • the availability of cloned granulins permit these questions to be addressed.
  • FIG. 4. shows the sequence of the probe used to screen a human bone marrow cDNA library in gtll (Clontech, Palo Alto, CA. ) .
  • the probe was generated by PCR of human DNA, with forward and reverse primers (underlined) based on the amino acid sequence of granulin A.
  • the nucleotides in lower case represent an intron in the coding region of granulin A.
  • Fig. 4B shows the sequencing strategy.
  • the nucleotide sequence is a composite of three sequences HBM12, HBM3 and HNM4.
  • Fig. 4C shows the complete nucleotide sequence and deduced polypeptide sequence of granulin. The nucleotides are numbered from the initiator codon (ATG), and the amino acids from the probable signal peptide cleavage site. Underlined sequences correspond to sequences previously determined by gas-phase microsequencing of purified granulins.
  • N-glycosylation sites are indicated by an asterisk (*) , and the stop codon is shown by a #.
  • the boxed nucleotides corresponds to a polyadenylation signal.
  • Clone HBM12 starts at nucleotide 6.
  • the sequences of clones HBM3 and HBM4 differ in the 5'-untranslated region as indicated by the bifurcation in the nucleotide sequence.
  • oligonucleotide primers forward primer, 5'- CGATGTGAAGTG(T/C)GA(T/C)ATGGA-3 ' ; reverse primer, 5'- CTGGCATGTGGTT(T/C)TC(A/G)CA(G/A)CA-3 ' ) were synthesized (Sheldon Biotechnology Centre, McGill University) and used in the polymerase chain reaction (PCR) with 2 ⁇ g of genomic DNA as template (34).
  • the amplified product was subcloned into the plasmid vector Bluescript KSII+ and its identity confirmed by double stranded dideoxynucleotide sequencing with Sequenase (USB). This fragment was labeled with 32 P by nick-translation (Boehringer Mannheim) and used to probe a human bone marrow cDNA library in ⁇ gtll (Cl ⁇ ntech, Palo Alto, CA. ) consisting of 1.51 X10 6 independent clones. Duplicate nitrocellulose filters (Schleicher and Schuell) were prehybridized in 5X SSC, 5X Denhardt's reagent, 0.2% SDS at 37°C for 5 hours.
  • Hybridization was in 5X SSC, 2.5X Denhardt's reagent, 0.2% SDS, 50% formamide, 10% polyethylene glycol with 1x10? cpm probe at 37°C for 12 hours. Filters were washed twice for 45 minutes in 2X SSC, 0.1% SDS at 58°C and exposed at -70°C with Kodak X-Omat film with an intensifying screen. 16 positives were obtain from 3x10 ⁇ clones screened. The cDNA insert from clone HMB12 was digested with Kpnl and Sad, and the resulting cDNA fragments subcloned into Bluescript KSII+.
  • Nucleotide sequence was determined by double stranded dideoxy sequencing using Sequenase (USB).
  • the sequence of HBM12 lacked an initiator methionine.
  • the remaining positive clones were analysed using PCR with ⁇ gtll sequence specific primers in combination with a primer, cll2rp, corresponding to nucleotides 45 to 61 of clone HBM12.
  • Fig.5. Comparison of the granulin-like domains. Residues occuring three or more times are boxed, and dashes have been introduced to align the cysteines.
  • the domain boundaries were determined by gas-phase microsequencing and by comparison with the sequence of grnA. The domains are located at amin ⁇ acids, 264-319, grnA; 169-244, grnB; 347-400, 425-479, grnD; 501-506, grnE; 106-162, grnF; 4i grnG and 1-27, paragranulin.
  • the order of domains is: paragranulin-G-F-B-A-C-D-E. They are not alphabetically aligned because several domains were isolated and named as discrete proteins before their common origin was known.
  • Fig. 7 Northern blot analysis of: A. granulin precursor mRNA in the following cell lines,
  • HL-60 promycelocytic leukemia
  • U937 histocytic leukemia
  • K562 histocytic leukemia
  • KMOE proerythroid leuker ⁇ ias
  • A431 epidermoid carcinoma
  • A54 lung epithelial
  • RNA from A431 (Chinese hamster ovaries), and murine Balb/C 3T3 fibroblasts, exposed for 10 days. (B). RNA from A431,
  • RNA was denatured with glyoxal, electrophoresed on a 1.1% agarose gel in 10 mM NaH2P ⁇ 4, pH 6.8, transferred to nylon membranes (ZetaProbe, BioRad) by capillary blotting with lOmM NaOH and fixed by baking at 80°C for 2 hours.
  • the membranes were hybridized at 65°C for RNA extracted from cell lines or 60°C for rabbit tissue RNA in 0.5M NaH 2 P ⁇ 4, pH 6.8, 7% SDS, lmM EDTA for 24 hours with the same probe described in Figure 6.
  • the membranes were washed at hybridization temperature for 30 mins. in 40 mM NaH2P ⁇ , pH 6.8, 5%
  • VIHCDAATICPDGTICCLSPYGMBGQCCRDGIHCCRHGYHCDSRTTHCL was isolated and characterized. It was found to be homologous with rat and human granulins but not identical.
  • the carp Granulin causes an increase in ⁇ E- thymidine incorporation in a keratinocyte cell line which indicates increased cell growth in keratinocytes and thus wound healing.
  • Granulin A causes a fall in 3n-t,hymidine incorporation in A-431 cells, a human epidermal carcinoma cell line.
  • the human granulins of the invention retard growth of A-431 cells.
  • the granulins of the invention are suitably employed as the active ingredient in pharmaceutical compositions for topical application.
  • the granulins may be incorporated in a topical formulation in an amount of 5 to 50 ⁇ g/ml of
  • SUBSTITUTE SHEET ointment and are applied to a wound site in an amount of 5 to 50 ⁇ g/cm 2 of wound site.
  • A. preferred treatment amount of the granulins is about lO ⁇ g/cm 2 of wound site suitably applied three times a day.
  • the topical pharmaceutical composition may be in the form of a cream, ointment, gel, lotion or other formulation suitable for application to the skin, and thus comprise a granulin of the invention admixed with an acceptable carrier for topical application.
  • SUBSTITUTESHEET TABLE 1 The amino acid compositions of purified granulins. The recovery of cysteine in this system is variable and is between 65 to 80%. Values have not been corrected for background contamination or oxidation. Tryptophan was not determine. Predicted values for granulin A from the gas-phase sequence determinations are given in brackets.
  • SUBSTITUTE SHEET TABLE 2 Structural analyses of five members of the granulin family.
  • the proposed structure for granulin A was determined by overlapping two amino terminal sequences, (1-11), and (1-23); tryptic fragments (4- 18), (19-52), and V8 protease fragment (37-56), and chymotryptic fragment (47-56).
  • the proposed structure of rat granulin is based partly upon the direct sequencing of the peptide itself (i.e., the sequence 1 through 21) and its proteolytic fragments (i.e., fragments corresponding to 4 to 18, 19 to 31, 32 to 46, and 32 to 51 sequences).
  • Rat Granulin EVKCDLEVSCPDGYTCCRLNTGAWG(CCPFSB)AVCCE-
  • Granulin B VMCPDARSRCPDGHTCCELPSGKYGCCPMPNATCCSDH- LHCCPQDTVCDLIQSKCI.
  • Granulin C VPCDXVSSCPSSDTCCOLTSGEHGCCPIPEAVC.
  • Granulin D IGCDQXDTSSCCPDG.
  • TFA trifluoroacetic acid: HFBA, heptafluorobutyric acid; rp-HPLC reverses phase high performance liquid chromatography; PTH, phenylthiohydantoin.
  • Cys Cys Pro lie Pro Glu Ala Val Cys Cys Ser 25 30 35 Asp His Gin His Cys Cys Pro Gin Arg Tyr Thr Cys
  • position 31 is not yet determined; it is either Asp or Asn.
  • identity of position 49 is not known.
  • position 23 is not yet determine; it is either Asp or Asn.

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Abstract

De nouveaux peptides de leucocytes peuvent être utilisés pour la cicatrisation des blessures; ces peptides sont riches en cystine et présentent une masse moléculaire de 6 Kda environ.
PCT/CA1992/000089 1992-02-03 1992-02-28 Granulines obtenues des leucocytes WO1993015195A1 (fr)

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Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6720159B1 (en) * 1997-12-16 2004-04-13 A&G Pharmaceutical, Inc. 88KDA tumorigenic growth factor and antagonists
US6872703B2 (en) 2000-06-07 2005-03-29 Ajinomoto Co., Inc. Insulin receptor-related receptor binding protein
US7091047B2 (en) 1997-05-23 2006-08-15 A&G Pharmaceutical, Inc. Methods and kits for diagnosing tumorigenicity
WO2009010045A1 (fr) * 2007-07-16 2009-01-22 Johann Wolfgang Goethe-Universität Frankfurt am Main Utilisation d'une granuline ou d'un composé analogue à la granuline pour la thérapie ou la prophylaxie des douleurs chroniques
US7651854B2 (en) 2003-02-26 2010-01-26 A & G Pharmaceutical, Inc. Methods for increasing the proliferation of B cells
US20100111928A1 (en) * 1997-05-23 2010-05-06 A & G Pharmaceutical, Inc. Methods and kits for diagnosis tumorgenicity
US7815906B2 (en) 2003-08-01 2010-10-19 A & G Pharmaceutical, Inc. Compositions and methods for restoring sensitivity to treatment with HER2 antagonists
US8088373B2 (en) 2002-11-19 2012-01-03 A&G Pharmaceutical, Inc. Autocrine growth factor receptor antibodies and methods
US8911950B2 (en) 1997-05-23 2014-12-16 A&G Pharmaceutical, Inc. Methods and compositions for inhibiting the growth of hematopoietic malignant cells
JP2014238343A (ja) * 2013-06-07 2014-12-18 株式会社島津製作所 質量分析装置を用いたプログラニュリンおよびグラニュリンペプチドの定量分析方法、および分析用プログラム
US8999365B2 (en) 2005-02-01 2015-04-07 Sinclair Pharmaceuticals Limited Prevention of bacterial contamination
WO2018039748A1 (fr) 2016-09-02 2018-03-08 James Cook University Peptide de cicatrisation de plaie
CN110903377A (zh) * 2019-11-08 2020-03-24 上海交通大学 一种生物活性多肽igcdqhtscpvgqtccps及其制备方法和应用

Citations (1)

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Publication number Priority date Publication date Assignee Title
WO1991015510A1 (fr) * 1990-04-03 1991-10-17 Bristol-Myers Squibb Company Epithelines: de nouvelles proteines modulatrices de la croissance riches en cysteine

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WO1991015510A1 (fr) * 1990-04-03 1991-10-17 Bristol-Myers Squibb Company Epithelines: de nouvelles proteines modulatrices de la croissance riches en cysteine

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Title
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS vol. 173, no. 3, 31 December 1990, DULUTH, MINNESOTA US pages 1161 - 1168 BATEMAN, A. ET AL. 'Granulins, a novel class of peptide from leukocytes' cited in the application *
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA vol. 87, no. 20, October 1990, WASHINGTON US pages 7912 - 7916 SHOYAB, M. ET AL. 'Epithelins 1 and 2: Isolation and characterization of two cysteine-rich growth-modulating proteins' cited in the application *

Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7091047B2 (en) 1997-05-23 2006-08-15 A&G Pharmaceutical, Inc. Methods and kits for diagnosing tumorigenicity
US20100111928A1 (en) * 1997-05-23 2010-05-06 A & G Pharmaceutical, Inc. Methods and kits for diagnosis tumorgenicity
US8911950B2 (en) 1997-05-23 2014-12-16 A&G Pharmaceutical, Inc. Methods and compositions for inhibiting the growth of hematopoietic malignant cells
US6720159B1 (en) * 1997-12-16 2004-04-13 A&G Pharmaceutical, Inc. 88KDA tumorigenic growth factor and antagonists
US6872703B2 (en) 2000-06-07 2005-03-29 Ajinomoto Co., Inc. Insulin receptor-related receptor binding protein
US8088373B2 (en) 2002-11-19 2012-01-03 A&G Pharmaceutical, Inc. Autocrine growth factor receptor antibodies and methods
US7651854B2 (en) 2003-02-26 2010-01-26 A & G Pharmaceutical, Inc. Methods for increasing the proliferation of B cells
US7815906B2 (en) 2003-08-01 2010-10-19 A & G Pharmaceutical, Inc. Compositions and methods for restoring sensitivity to treatment with HER2 antagonists
US8999365B2 (en) 2005-02-01 2015-04-07 Sinclair Pharmaceuticals Limited Prevention of bacterial contamination
DE102007033359A1 (de) * 2007-07-16 2009-01-29 Johann Wolfgang Goethe-Universität Frankfurt am Main Verwendung eines Granulins oder einer Granulin-ähnlichen Verbindung zur Therapie oder Prophylaxe von chronischen Schmerzen
DE102007033359B4 (de) * 2007-07-16 2010-12-02 Johann Wolfgang Goethe-Universität Frankfurt am Main Verwendung eines Granulins oder einer Granulin-ähnlichen Verbindung zur Therapie oder Prophylaxe von chronischen Schmerzen
US8367616B2 (en) 2007-07-16 2013-02-05 Johann Wolfgang Goethe-Universitat Frankfurt Am Main Use of a granulin or a granulin-like compound in the therapy or prophylaxis of chronic pains
JP2010533660A (ja) * 2007-07-16 2010-10-28 ヨハン ウォルフガング ゲーテ−ウニベルジテート フランクフルト アム マイン 慢性疼痛の治療又は予防のためのグラニュリン又はグラニュリン様化合物の使用
WO2009010045A1 (fr) * 2007-07-16 2009-01-22 Johann Wolfgang Goethe-Universität Frankfurt am Main Utilisation d'une granuline ou d'un composé analogue à la granuline pour la thérapie ou la prophylaxie des douleurs chroniques
US9655947B2 (en) 2007-07-16 2017-05-23 Johann Wolfgang Goethe-Universitat Frankfurt Am Main Use of a granulin or a granulin-like compound for the therapy or prophylaxis of chronic pain
JP2014238343A (ja) * 2013-06-07 2014-12-18 株式会社島津製作所 質量分析装置を用いたプログラニュリンおよびグラニュリンペプチドの定量分析方法、および分析用プログラム
WO2018039748A1 (fr) 2016-09-02 2018-03-08 James Cook University Peptide de cicatrisation de plaie
EP3507300A4 (fr) * 2016-09-02 2020-04-08 James Cook University Peptide de cicatrisation de plaie
CN110903377A (zh) * 2019-11-08 2020-03-24 上海交通大学 一种生物活性多肽igcdqhtscpvgqtccps及其制备方法和应用

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