DOCSLIB.ORG

(12) Patent Application Publication (10) Pub. No.: US 2011/0044968 A1 Bolotin Et Al

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
(12) Patent Application Publication (10) Pub. No.: US 2011/0044968 A1 Bolotin Et Al US 20110044968A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0044968 A1 Bolotin et al. (43) Pub. Date: Feb. 24, 2011 (54) COMPOSITIONS FOR TREATMENT WITH (86). PCT No.: PCT/USO9A36648 METALLOPEPTIDASES, METHODS OF MAKING AND USING THE SAME S371 (c)(1), (2), (4) Date: Nov. 5, 2010 (75) Inventors: Elijah M. Bolotin, Bothell, WA Related U.S. Application Data WA(US); (US); Gerardo Penelope Castillo, Markham, Bothell, (60) Fyal application No. 61/068.896, filed on Mar. Clifton, VA (US); Manshun Lai, s Bothell, WA (US) Publication Classification (51) Int. Cl. Correspondence Address: A638/54 (2006.01) WILSON SONSIN GOODRCH AND ROSAT f CI2N 9/96 (2006.01) PHARMAN LTD A6IP3L/00 (2006.01) 650 PAGE MILL ROAD A6IP 25/28 (2006.01) PALO ALTO, CA 94.304 (US) (52) U.S. Cl. ........................................ 424/94.3; 435/188 (57) ABSTRACT (73)73) AssigneeAssi : WPh re NC orporation,tion. SeattlSealue, The present invention is directed to biocompatible composi tions and the use of metal bridges to connect a back-bone and a metallopeptidase active agent. In certain instances, the Sub (21) Appl. No.: 12/921,670 ject compositions provide a means of achieving Sustained release of the metallopeptidase active agent after administra (22) PCT Filed: Mar. 10, 2009 tion to a subject. Patent Application Publication Feb. 24, 2011 Sheet 1 of 13 US 2011/0044968 A1 Figure 1 (1) Polymeric Backbone (branched or unbranched) 2) Chelating moieties (3) Metalions ( 4) Protective chains (5) Metallopeptidase active agent Patent Application Publication Feb. 24, 2011 Sheet 2 of 13 US 2011/0044968 A1 Figure 2 O 120% poss C D 100% O 80% w 60% s CD 20% n S PGC-NTA-Zn PGC-NTA Lysostaphin Lysostaphin Patent Application Publication Feb. 24, 2011 Sheet 3 of 13 US 2011/0044968 A1 Figure 3 100 Bound (uM) Patent Application Publication Feb. 24, 2011 Sheet 4 of 13 US 2011/0044968 A1 Figure 4 100 cy Bound (uM) Patent Application Publication Feb. 24, 2011 Sheet 5 of 13 US 2011/0044968 A1 Figure 5 8 O A.6 OO 2. O 11 12 13 14 15 16 Bound (uM) Patent Application Publication Feb. 24, 2011 Sheet 6 of 13 US 2011/0044968 A1 Figure 6 Lot # 20080421a, 20080326, and 20080421 a 50% loading as a Lysostaphin Alone to 20PLPEG1055DAPE4NTAZn a. 10 1 Sks vs. krrv.rssssssssssssss . krrrrrrrrrrrrrrrrrrrrrrr . is rrrrrrrrrrrrrr's . V 5 E 1 - SRs Nasses . sesssssssssssssssssssssssssss s g W 0.1 rarr rrrrrr x x . wSvrr\ rrrrrrrrrrrrrrrra s vs. xxxx xxx's SasssssssserN S. Y N s sa s w y ‘ a n x.S. s. s. NXX w 0.01 sys O 20 40 60 80 100 Time (h) Patent Application Publication Feb. 24, 2011 Sheet 7 of 13 US 2011/0044968 A1 Figure 7 Lot #20080603c and 20080804b 50% loading 1000 “NLysostaphi Alone “ i W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. W. Cu2OPPEG550DAPE4NTAZn. N 18PEIPEG103ODANTAZn wars sax sas sas SSS Patent Application Publication Feb. 24, 2011 Sheet 8 of 13 US 2011/0044968 A1 Figure 8 Lot #20080603c, 20080604c, and 20080605c 20% loading 1OOO “was lysostaphin Alone “ 100 user2OPLPEG550DAPEIANTAZn. 2OPLPEG55ODAPE8NTAZ 10 S - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - missix 20PLPEG55ODAPE12NTAZn Patent Application Publication Feb. 24, 2011 Sheet 9 of 13 US 2011/0044968 A1 Figure 9 SucNTA SucNTA ZnNTA ZnNTA NiNTA NiNTA 2 mg 1 mg 2 mg 1 mg 2 mg Patent Application Publication Feb. 24, 2011 Sheet 10 of 13 US 2011/0044968 A1 Figure 10 3 O OOOO Column 1 2 OOOOO E L L 1 OOOOO Column 2 O HHHO-HO- HO- PDO O 1 O 2O Fraction number Patent Application Publication Feb. 24, 2011 Sheet 11 of 13 US 2011/0044968 A1 Figure 11 PEGNiNTAthplasia PENNA PEGarul A-plasia PEN PEG-PL-plasma PEG-PL PEG-PL GFPH O 15 GFP Oud Patent Application Publication Feb. 24, 2011 Sheet 12 of 13 US 2011/0044968 A1 Figure 12 th. areas 6 3. 3 Control 2. MPEG-PL-ZnNTA MPEG-PL-NiNTA 1. tire, air Patent Application Publication Feb. 24, 2011 Sheet 13 of 13 US 2011/0044968 A1 Figure 13 y Š Accumulation in infanation site (rat model US 2011/0044968 A1 Feb. 24, 2011 COMPOSITIONS FOR TREATMENT WITH longer circulation in the body, more stability in the blood, and METALLOPEPTIDASES, METHODS OF can be more conveniently administered. MAKING AND USING THE SAME SUMMARY OF THE INVENTION CROSS-REFERENCE 0006. In part, the present invention is directed to biocom patible compositions and use of metal bridges to connect a 0001. This application claims the benefit of U.S. Provi backbone and a metallopeptidase active agent. In certain sional Application No. 61/068,896 filed Mar. 10, 2008, which instances, the Subject compositions provide a means of application is incorporated herein by reference in its entirety. achieving Sustained release of the metallopeptidase active agent after administration to a Subject. The metallopeptidase STATEMENT AS TO FEDERALLY SPONSORED can be one that is selected from those listed in Table 1 or Table 2. In one embodiment, the metallopeptidase active agent is a RESEARCH metalloexopeptidase or a metallocarboxypeptidase. In 0002 This invention was made with the support of the another embodiment, the metallopeptidase active agent is a U.S. Government under Grant No. 5R43AIO78539 awarded metalloendopeptidase. In a specific embodiment, the metal by the National Institute of Allergy and Infectious Disease lopeptidase active agent is a glycyl-glycyl metalloendopepti (NIAID). The U.S. Government may have certain rights to the dase, such as lysostaphin. In another specific embodiment, subject matter provided herein. the metallopeptidase active agent is the metalloendopepti dase neprilysin. 0007. In one aspect, the present invention relates compo BACKGROUND OF THE INVENTION sition containing (i) a polymeric backbone with monomeric units, (ii) a chelating group covalently linked to a monomeric 0003. The development of new drugs, formulations and unit, (iii) a transition metal ion and iv) a metallopeptidase other systems for administration of physiologically active active agent coordinately bonded to the transition metal ion. peptides and proteins and other therapeutics and materials is In another aspect, the present invention relates to a composi driven by the need to provide these peptides or proteins or tion comprising (i) an aliphatic group, (ii) a chelating group other materials to achieve the desirable physiological effects. covalently linked to the aliphatic group, (iii) a transition metal With respect to peptides and proteins, many of them have ion, and iv) a metallopeptidase active agent coordinately been observed to be unstable in the gastro-intestinal tract and bonded to the transition metalion. therefore may need to be stabilized or protected or delivered 0008. The polymeric backbone of the subject composi via systemic circulation. In addition, peptides and proteins tions can be chosen from but not limited to polylysine, pol that have low molecular masses tend to have short biological yaspartic acid, polyglutamic acid, polyserine, polythreonine, half-lives due to their efficient removal from systemic circu polycysteine, polyglycerol, polyethyleneimines, polyally lation via kidneys and reticuloendothelial system. Many pep lamine, chitosan, natural saccharides, aminated polysaccha tides and proteins can also lose their activity in vivo due to rides, aminated oligosaccharides, polyamidoamine, poly proteolysis (peptide bond cleavage). acrylic acids, polyalcohols, Sulfonated polysaccharides, 0004. In part to circumvent these undesirable effects, a Sulfonated oligosaccharides, carboxylated polysaccharides, drug delivery system may be used. Drug delivery strategies carboxylated oligosaccharides, aminocarboxylated polysac have been developed for peptide and protein delivery in vivo, charides, aminocarboxylated oligosaccharides, carboxym but most are not useful for sustained delivery. For example, ethylated polysaccharides, or carboxymethylated oligosac the use of a continuous systemic infusion of drug via a pump charides. is impractical for outpatients requiring high levels of mobility 0009. The aliphatic chain of the subject compositions can and has the associated disadvantages of quality of life and be within a general formula PvNwCxHyOZ- where v is 0-3, potential intravenous (I.V.) line infections. The use of an w is 0-3, X is 8–48; y is 15-95; Z is 1-13. In a further embodi implantable pump, comprised of a capsule with a membrane ment of the above compositions with an aliphatic group, the allowing diffusion of a drug, is limited by the volume of the aliphatic group is an alkyl group. In one embodiment the capsule. Peptides and proteins are often used in concentrated aliphatic chain comprises from C8 to C36 carbon atoms formulations in the capsules and aggregate, whereby losing inclusive. In a further embodiment, the alkyl group comprises specific activity. In many cases, the drug is released into the a general formula CH (CH)x- where x is 5-35. In a further extracellular space and distributed in lymphatics. Other embodiment, the aliphatic group comprises one or more alkyl implantable biodegradable delivery systems are implanted or group(s) derived from various fatty acids or fatty acids with injected into the epidermis. The components of the system are aromatic group(s). In further embodiments, the aliphatic usually slowly degraded as a result of biological activity of group is within the structure that comprises phospholipids or Surrounding cells (i.e. as a result of the release of enzymes derivative of phospholipids. In further embodiments, the ali degrading chemical bonds that hold these implants together). phatic group is within the structure that comprises diacylg 0005 Metallopeptidases, interchangeably known as met lycerol or derivatives of diacylglycerol. In a further embodi alloproteinases and metalloproteases, encompass a large fam ment, the alkyl group comprises a branched alkyl group.
Load more

Recommended publications
  • Proteomic Analysis of the Venom of Jellyfishes Rhopilema Esculentum and Sanderia Malayensis
    marine drugs Article Proteomic Analysis of the Venom of Jellyfishes Rhopilema esculentum and Sanderia malayensis 1, 2, 2 2, Thomas C. N. Leung y , Zhe Qu y , Wenyan Nong , Jerome H. L. Hui * and Sai Ming Ngai 1,* 1 State Key Laboratory of Agrobiotechnology, School of Life Sciences, The Chinese University of Hong Kong, Hong Kong, China; [email protected] 2 Simon F.S. Li Marine Science Laboratory, State Key Laboratory of Agrobiotechnology, School of Life Sciences, The Chinese University of Hong Kong, Hong Kong, China; [email protected] (Z.Q.); [email protected] (W.N.) * Correspondence: [email protected] (J.H.L.H.); [email protected] (S.M.N.) Contributed equally. y Received: 27 November 2020; Accepted: 17 December 2020; Published: 18 December 2020 Abstract: Venomics, the study of biological venoms, could potentially provide a new source of therapeutic compounds, yet information on the venoms from marine organisms, including cnidarians (sea anemones, corals, and jellyfish), is limited. This study identified the putative toxins of two species of jellyfish—edible jellyfish Rhopilema esculentum Kishinouye, 1891, also known as flame jellyfish, and Amuska jellyfish Sanderia malayensis Goette, 1886. Utilizing nano-flow liquid chromatography tandem mass spectrometry (nLC–MS/MS), 3000 proteins were identified from the nematocysts in each of the above two jellyfish species. Forty and fifty-one putative toxins were identified in R. esculentum and S. malayensis, respectively, which were further classified into eight toxin families according to their predicted functions. Amongst the identified putative toxins, hemostasis-impairing toxins and proteases were found to be the most dominant members (>60%).
    [Show full text]
  • Regulation of Procollagen Amino-Propeptide Processing During Mouse Embryogenesis by Specialization of Homologous ADAMTS Protease
    DEVELOPMENT AND DISEASE RESEARCH ARTICLE 1587 Development 133, 1587-1596 (2006) doi:10.1242/dev.02308 Regulation of procollagen amino-propeptide processing during mouse embryogenesis by specialization of homologous ADAMTS proteases: insights on collagen biosynthesis and dermatosparaxis Carine Le Goff1, Robert P. T. Somerville1, Frederic Kesteloot2, Kimerly Powell1, David E. Birk3, Alain C. Colige2 and Suneel S. Apte1,* Mutations in ADAMTS2, a procollagen amino-propeptidase, cause severe skin fragility, designated as dermatosparaxis in animals, and a subtype of the Ehlers-Danlos syndrome (dermatosparactic type or VIIC) in humans. Not all collagen-rich tissues are affected to the same degree, which suggests compensation by the ADAMTS2 homologs ADAMTS3 and ADAMTS14. In situ hybridization of Adamts2, Adamts3 and Adamts14, and of the genes encoding the major fibrillar collagens, Col1a1, Col2a1 and Col3a1, during mouse embryogenesis, demonstrated distinct tissue-specific, overlapping expression patterns of the protease and substrate genes. Adamts3, but not Adamts2 or Adamts14, was co-expressed with Col2a1 in cartilage throughout development, and with Col1a1 in bone and musculotendinous tissues. ADAMTS3 induced procollagen I processing in dermatosparactic fibroblasts, suggesting a role in procollagen I processing during musculoskeletal development. Adamts2, but not Adamts3 or Adamts14, was co-expressed with Col3a1 in many tissues including the lungs and aorta, and Adamts2–/– mice showed widespread defects in procollagen III processing. Adamts2–/– mice had abnormal lungs, characterized by a decreased parenchymal density. However, the aorta and collagen fibrils in the aortic wall appeared normal. Although Adamts14 lacked developmental tissue-specific expression, it was co-expressed with Adamts2 in mature dermis, which possibly explains the presence of some processed skin procollagen in dermatosparaxis.
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 6,395,889 B1 Robison (45) Date of Patent: May 28, 2002
    USOO6395889B1 (12) United States Patent (10) Patent No.: US 6,395,889 B1 Robison (45) Date of Patent: May 28, 2002 (54) NUCLEIC ACID MOLECULES ENCODING WO WO-98/56804 A1 * 12/1998 ........... CO7H/21/02 HUMAN PROTEASE HOMOLOGS WO WO-99/0785.0 A1 * 2/1999 ... C12N/15/12 WO WO-99/37660 A1 * 7/1999 ........... CO7H/21/04 (75) Inventor: fish E. Robison, Wilmington, MA OTHER PUBLICATIONS Vazquez, F., et al., 1999, “METH-1, a human ortholog of (73) Assignee: Millennium Pharmaceuticals, Inc., ADAMTS-1, and METH-2 are members of a new family of Cambridge, MA (US) proteins with angio-inhibitory activity', The Journal of c: - 0 Biological Chemistry, vol. 274, No. 33, pp. 23349–23357.* (*) Notice: Subject to any disclaimer, the term of this Descriptors of Protease Classes in Prosite and Pfam Data patent is extended or adjusted under 35 bases. U.S.C. 154(b) by 0 days. * cited by examiner (21) Appl. No.: 09/392, 184 Primary Examiner Ponnathapu Achutamurthy (22) Filed: Sep. 9, 1999 ASSistant Examiner William W. Moore (51) Int. Cl." C12N 15/57; C12N 15/12; (74) Attorney, Agent, or Firm-Alston & Bird LLP C12N 9/64; C12N 15/79 (57) ABSTRACT (52) U.S. Cl. .................... 536/23.2; 536/23.5; 435/69.1; 435/252.3; 435/320.1 The invention relates to polynucleotides encoding newly (58) Field of Search ............................... 536,232,235. identified protease homologs. The invention also relates to 435/6, 226, 69.1, 252.3 the proteases. The invention further relates to methods using s s s/ - - -us the protease polypeptides and polynucleotides as a target for (56) References Cited diagnosis and treatment in protease-mediated disorders.
    [Show full text]
  • Repression of Anti-Proliferative Factor Tob1 in Osteoarthritic Cartilage
    Available online http://arthritis-research.com/content/7/2/R274 ResearchVol 7 No 2 article Open Access Repression of anti-proliferative factor Tob1 in osteoarthritic cartilage Mathias Gebauer1*, Joachim Saas2*, Jochen Haag3, Uwe Dietz2, Masaharu Takigawa4, Eckart Bartnik2 and Thomas Aigner3 1Aventis Pharma Deutschland, Functional Genomics, Sanofi-Aventis, Frankfurt, Germany 2Sanofi-Aventis, Disease Group Thrombotic Diseases/Degenerative Joint Diseases, Frankfurt, Germany 3Osteoarticular and Arthritis Research, Department of Pathology, University of Erlangen-Nürnberg, Germany 4Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine and Dentistry, Okayama, Japan * Contributed equally Corresponding author: Thomas Aigner, [email protected] Received: 10 Aug 2004 Revisions requested: 1 Oct 2004 Revisions received: 22 Oct 2004 Accepted: 19 Nov 2004 Published: 11 Jan 2005 Arthritis Res Ther 2005, 7:R274-R284 (DOI 10.1186/ar1479)http://arthritis-research.com/content/7/2/R274 © 2005 Gebauer et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/ 2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Osteoarthritis is the most common degenerative disorder of the genes were detected between normal and osteoarthritic modern world. However, many basic cellular features and cartilage (P < 0.01). One of the significantly repressed genes, molecular processes of the disease are poorly understood. In Tob1, encodes a protein belonging to a family involved in the present study we used oligonucleotide-based microarray silencing cells in terms of proliferation and functional activity.
    [Show full text]
  • The Astacin Family of Metalloendopeptidases
    Profein Science (1995), 4:1247-1261. Cambridge University Press. Printed in the USA Copyright 0 1995 The Protein Society The astacin family of metalloendopeptidases JUDITH S. BOND’ AND ROBERT J. BEYNON2 ’ Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033 Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute of ‘Science and Technology, Manchester M60 1QD. United Kingdom (RECEIVEDMarch 23, 1995; ACCEPTED April19, 1995) Abstract The astacin family of metalloendopeptidases was recognized as a novel family of proteases in the 1990s. The cray- fish enzyme astacin was the first characterized and is one of the smallest members of the family. More than 20 members of the family have nowbeen identified. They have been detected in species ranging from hydra to hu- mans, in mature andin developmental systems. Proposed functions of these proteases include activation of growth factors, degradation of polypeptides, and processing of extracellular proteins. Astacin family proteases aresyn- thesized with NH,-terminal signal and proenzyme sequences, and many (such as meprins, BMP-1, folloid)con- tain multiple domains COOH-terminal to the protease domain. They are eithersecreted from cells or are plasma membrane-associated enzymes. They have some distinguishing features in addition to the signature sequencein the protease domain: HEXXHXXGFXHEXXRXDR. They have a unique typeof zinc binding, with pentacoor- dination, and a protease domain tertiary structure that contains common attributeswith serralysins, matrix me- talloendopeptidases, and snake venom proteases; they cleave peptide bonds in polypeptides such as insulin B chain and bradykinin andin proteins such as casein and gelatin; and theyhave arylamidase activity.
    [Show full text]
  • Structural Basis for the Sheddase Function of Human Meprin Β Metalloproteinase at the Plasma Membrane
    Structural basis for the sheddase function of human meprin β metalloproteinase at the plasma membrane Joan L. Arolasa, Claudia Broderb, Tamara Jeffersonb, Tibisay Guevaraa, Erwin E. Sterchic, Wolfram Boded, Walter Stöckere, Christoph Becker-Paulyb, and F. Xavier Gomis-Rütha,1 aProteolysis Laboratory, Department of Structural Biology, Molecular Biology Institute of Barcelona, Consejo Superior de Investigaciones Cientificas, Barcelona Science Park, E-08028 Barcelona, Spain; bInstitute of Biochemistry, Unit for Degradomics of the Protease Web, University of Kiel, D-24118 Kiel, Germany; cInstitute of Biochemistry and Molecular Medicine, University of Berne, CH-3012 Berne, Switzerland; dArbeitsgruppe Proteinaseforschung, Max-Planck-Institute für Biochemie, D-82152 Planegg-Martinsried, Germany; and eInstitute of Zoology, Cell and Matrix Biology, Johannes Gutenberg-University, D-55128 Mainz, Germany Edited by Brian W. Matthews, University of Oregon, Eugene, OR, and approved August 22, 2012 (received for review June 29, 2012) Ectodomain shedding at the cell surface is a major mechanism to proteolysis” step within the membrane (1). This is the case for the regulate the extracellular and circulatory concentration or the processing of Notch ligand Delta1 and of APP, both carried out by activities of signaling proteins at the plasma membrane. Human γ-secretase after action of an α/β-secretase (11), and for signal- meprin β is a 145-kDa disulfide-linked homodimeric multidomain peptide peptidase, which removes remnants of the secretory pro- type-I membrane metallopeptidase that sheds membrane-bound tein translocation from the endoplasmic membrane (13). cytokines and growth factors, thereby contributing to inflammatory Recently, human meprin β (Mβ) was found to specifically pro- diseases, angiogenesis, and tumor progression.
    [Show full text]
  • ADAMTS13 and 15 Are Not Regulated by the Full Length and N‑Terminal Domain Forms of TIMP‑1, ‑2, ‑3 and ‑4
    BIOMEDICAL REPORTS 4: 73-78, 2016 ADAMTS13 and 15 are not regulated by the full length and N‑terminal domain forms of TIMP‑1, ‑2, ‑3 and ‑4 CENQI GUO, ANASTASIA TSIGKOU and MENG HUEE LEE Department of Biological Sciences, Xian Jiaotong-Liverpool University, Suzhou, Jiangsu 215123, P.R. China Received June 29, 2015; Accepted July 15, 2015 DOI: 10.3892/br.2015.535 Abstract. A disintegrin and metalloproteinase with thom- proteolysis activities associated with arthritis, morphogenesis, bospondin motifs (ADAMTS) 13 and 15 are secreted zinc angiogenesis and even ovulation [as reviewed previously (1,2)]. proteinases involved in the turnover of von Willebrand factor Also known as the VWF-cleaving protease, ADAMTS13 and cancer suppression. In the present study, ADAMTS13 is noted for its ability in cleaving and reducing the size of the and 15 were subjected to inhibition studies with the full-length ultra-large (UL) form of the VWF. Reduction in ADAMTS13 and N-terminal domain forms of tissue inhibitor of metallo- activity from either hereditary or acquired deficiency causes proteinases (TIMPs)-1 to -4. TIMPs have no ability to inhibit accumulation of UL-VWF multimers, platelet aggregation and the ADAMTS proteinases in the full-length or N-terminal arterial thrombosis that leads to fatal thrombotic thrombocy- domain form. While ADAMTS13 is also not sensitive to the topenic purpura [as reviewed previously (1,3)]. By contrast, hydroxamate inhibitors, batimastat and ilomastat, ADAMTS15 ADAMTS15 is a potential tumor suppressor. Only a limited app can be effectively inhibited by batimastat (Ki 299 nM). In number of in-depth investigations have been carried out on the conclusion, the present results indicate that TIMPs are not the enzyme; however, expression and profiling studies have shown regulators of these two ADAMTS proteinases.
    [Show full text]
  • Serine Proteases with Altered Sensitivity to Activity-Modulating
    (19) & (11) EP 2 045 321 A2 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 08.04.2009 Bulletin 2009/15 C12N 9/00 (2006.01) C12N 15/00 (2006.01) C12Q 1/37 (2006.01) (21) Application number: 09150549.5 (22) Date of filing: 26.05.2006 (84) Designated Contracting States: • Haupts, Ulrich AT BE BG CH CY CZ DE DK EE ES FI FR GB GR 51519 Odenthal (DE) HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI • Coco, Wayne SK TR 50737 Köln (DE) •Tebbe, Jan (30) Priority: 27.05.2005 EP 05104543 50733 Köln (DE) • Votsmeier, Christian (62) Document number(s) of the earlier application(s) in 50259 Pulheim (DE) accordance with Art. 76 EPC: • Scheidig, Andreas 06763303.2 / 1 883 696 50823 Köln (DE) (71) Applicant: Direvo Biotech AG (74) Representative: von Kreisler Selting Werner 50829 Köln (DE) Patentanwälte P.O. Box 10 22 41 (72) Inventors: 50462 Köln (DE) • Koltermann, André 82057 Icking (DE) Remarks: • Kettling, Ulrich This application was filed on 14-01-2009 as a 81477 München (DE) divisional application to the application mentioned under INID code 62. (54) Serine proteases with altered sensitivity to activity-modulating substances (57) The present invention provides variants of ser- screening of the library in the presence of one or several ine proteases of the S1 class with altered sensitivity to activity-modulating substances, selection of variants with one or more activity-modulating substances. A method altered sensitivity to one or several activity-modulating for the generation of such proteases is disclosed, com- substances and isolation of those polynucleotide se- prising the provision of a protease library encoding poly- quences that encode for the selected variants.
    [Show full text]
  • Effect of Inhibition of Matrix Metalloproteinases on Endometrial Decidualization and Implantation in Mated Rats M
    Effect of inhibition of matrix metalloproteinases on endometrial decidualization and implantation in mated rats M. P. Rechtman, J. Zhang and L. A. Salamonsen Prince Henry's Institute ofMedical Research, PO Box 5152, Clayton, Victoria 3168, Australia Implantation of the embryo into the endometrium is a highly regulated event that is critical for establishment of pregnancy. Molecules involved in this process provide potential targets for post-coital contraception. The aims of this study were to determine whether matrix metalloproteinases (MMPs) are present at implantation sites in rats and whether administration of a broad-based inhibitor of MMPs could inhibit embryo implantation. Uterine extracts from non-pregnant rats and from rats on days 3\p=n-\9 of pregnancy were examined for the presence of MMPs. Doxycycline (5 or 15 mg day\m=-\1) was administered by gavage to rats from the day of mating (day 0) to day 7 of pregnancy and the uterus was examined for implantation sites. A number of MMPs were present in all uterine samples. MMP-2 reached a peak on day 3, whereas the highest expression of MMP-7 occurred on day 7. MMP-13 and MMP-3 were present in smaller amounts. MMP-9 was detectable only on day 9. Treatment of rats with doxycycline had no effect on the number of implantation sites or on the total uterine mass. However, in treated rats, the process of decidualization was impaired and both the width and length of the decidual zone was reduced, resulting in a decrease in total decidual area from 1.20 \m=+-\0.07 to 0.91 \m=+-\0.07 mm2 (mean \m=+-\sem, controls versus doxycycline treated, P < 0.02).
    [Show full text]
  • RSC Advances
    RSC Advances PAPER View Article Online View Journal | View Issue Experimental identification and computational characterization of a novel extracellular Cite this: RSC Adv.,2017,7,13928 metalloproteinase produced by Clostridium sordellii† Michael J. Aldape,*ab Aoxiang Tao,c Dustin D. Heeney,a Eric R. McIndoo,a John M. Frencha and Dong Xu*c Clostridium sordellii is a lethal pathogen for both animals and humans. Severe capillary leakage, toxic shock syndrome, and an extreme leukemoid reaction (LR), are hallmark features of C. sordellii infections and contribute to its high mortality rate. Here we report the discovery of a previously unknown and uncharacterized metalloproteinase of C. sordellii (referred as Mcs1) that cleaves human vascular cell adhesion molecule (VCAM)-1 in vitro, an adhesion molecule critical to hematopoietic precursor retention and leukocyte diapedesis. We successfully identified the open reading frame encoding Mcs1 within Creative Commons Attribution-NonCommercial 3.0 Unported Licence. the ATCC 9714 genome and developed an Dmcs1 mutant strain using the ClosTron mutagenesis technology. No VCAM-1 proteolysis was observed from exotoxins collected from mutant strain cultures. Using advanced protein structural modeling and molecular dynamics simulation techniques, the 3D molecular structure and conformational features of Mcs1 were also characterized. Our data demonstrates that Mcs1 proteolytic activity is controlled by the electrostatic interactions between Glu113 and Arg227 residues and the gating motions within its cleft region. This pilot interdisciplinary investigation provided crucial experimental evidence of the existence of Mcs1 in C. sordellii and Received 2nd December 2016 molecular insights into its 3D structure and proteolytic activity. These findings have the potential to help Accepted 22nd February 2017 This article is licensed under a advance new therapeutics and diagnostics against deadly C.
    [Show full text]
  • Supplementary Materials
    Supplementary materials Supplementary Table S1: MGNC compound library Ingredien Molecule Caco- Mol ID MW AlogP OB (%) BBB DL FASA- HL t Name Name 2 shengdi MOL012254 campesterol 400.8 7.63 37.58 1.34 0.98 0.7 0.21 20.2 shengdi MOL000519 coniferin 314.4 3.16 31.11 0.42 -0.2 0.3 0.27 74.6 beta- shengdi MOL000359 414.8 8.08 36.91 1.32 0.99 0.8 0.23 20.2 sitosterol pachymic shengdi MOL000289 528.9 6.54 33.63 0.1 -0.6 0.8 0 9.27 acid Poricoic acid shengdi MOL000291 484.7 5.64 30.52 -0.08 -0.9 0.8 0 8.67 B Chrysanthem shengdi MOL004492 585 8.24 38.72 0.51 -1 0.6 0.3 17.5 axanthin 20- shengdi MOL011455 Hexadecano 418.6 1.91 32.7 -0.24 -0.4 0.7 0.29 104 ylingenol huanglian MOL001454 berberine 336.4 3.45 36.86 1.24 0.57 0.8 0.19 6.57 huanglian MOL013352 Obacunone 454.6 2.68 43.29 0.01 -0.4 0.8 0.31 -13 huanglian MOL002894 berberrubine 322.4 3.2 35.74 1.07 0.17 0.7 0.24 6.46 huanglian MOL002897 epiberberine 336.4 3.45 43.09 1.17 0.4 0.8 0.19 6.1 huanglian MOL002903 (R)-Canadine 339.4 3.4 55.37 1.04 0.57 0.8 0.2 6.41 huanglian MOL002904 Berlambine 351.4 2.49 36.68 0.97 0.17 0.8 0.28 7.33 Corchorosid huanglian MOL002907 404.6 1.34 105 -0.91 -1.3 0.8 0.29 6.68 e A_qt Magnogrand huanglian MOL000622 266.4 1.18 63.71 0.02 -0.2 0.2 0.3 3.17 iolide huanglian MOL000762 Palmidin A 510.5 4.52 35.36 -0.38 -1.5 0.7 0.39 33.2 huanglian MOL000785 palmatine 352.4 3.65 64.6 1.33 0.37 0.7 0.13 2.25 huanglian MOL000098 quercetin 302.3 1.5 46.43 0.05 -0.8 0.3 0.38 14.4 huanglian MOL001458 coptisine 320.3 3.25 30.67 1.21 0.32 0.9 0.26 9.33 huanglian MOL002668 Worenine
    [Show full text]
  • Structural Basis of Mammalian Mucin Processing by the Human Gut O
    ARTICLE https://doi.org/10.1038/s41467-020-18696-y OPEN Structural basis of mammalian mucin processing by the human gut O-glycopeptidase OgpA from Akkermansia muciniphila ✉ ✉ Beatriz Trastoy 1,4, Andreas Naegeli2,4, Itxaso Anso 1,4, Jonathan Sjögren 2 & Marcelo E. Guerin 1,3 Akkermansia muciniphila is a mucin-degrading bacterium commonly found in the human gut that promotes a beneficial effect on health, likely based on the regulation of mucus thickness 1234567890():,; and gut barrier integrity, but also on the modulation of the immune system. In this work, we focus in OgpA from A. muciniphila,anO-glycopeptidase that exclusively hydrolyzes the peptide bond N-terminal to serine or threonine residues substituted with an O-glycan. We determine the high-resolution X-ray crystal structures of the unliganded form of OgpA, the complex with the glycodrosocin O-glycopeptide substrate and its product, providing a comprehensive set of snapshots of the enzyme along the catalytic cycle. In combination with O-glycopeptide chemistry, enzyme kinetics, and computational methods we unveil the molecular mechanism of O-glycan recognition and specificity for OgpA. The data also con- tribute to understanding how A. muciniphila processes mucins in the gut, as well as analysis of post-translational O-glycosylation events in proteins. 1 Structural Biology Unit, Center for Cooperative Research in Biosciences (CIC bioGUNE), Basque Research and Technology Alliance (BRTA), Bizkaia Technology Park, Building 801A, 48160 Derio, Spain. 2 Genovis AB, Box 790, 22007 Lund, Sweden. 3 IKERBASQUE, Basque Foundation for Science, 48013 ✉ Bilbao, Spain. 4These authors contributed equally: Beatriz Trastoy, Andreas Naegeli, Itxaso Anso.
    [Show full text]