Title (en)
Cloning, yeast expression, purification and biological activity of a truncated form of the soybean 7S globulin alfa ' subunit involved in Hep G2 cell
cholesterol homeostasis
Title (de)
Klonierung, Hefeexpression, Aufreinigung und biologische Aktivität einer verkürzten Form der Soja-7S-Globulin-alfa-Untereinheit, die an der Hep-G2Zellen-Cholesterin-Homöostase beteiligt ist
Title (fr)
Clonage, expression de levure, purification et activité biologique d'une formule tronquée de la sous-unité alpha 7S globuline de soja impliquée dans
l'homéostasie du cholestérol de cellule Hep G2
Publication
EP 2264054 A1 20101222 (EN)
Application
EP 09162935 A 20090617
Priority
EP 09162935 A 20090617
Abstract (en)
A truncated form of ±' chain (t±'), the soybean 7S globulin, active in controlling the cholesterol and triglyceride homeostasis in in vitro and in vivo
models, was cloned and expressed in the yeast Pichia pastoris. The recombinant polypeptide spanned 216 amino acid residues from the N-terminal
side and included the N-terminal extension region of the soybean subunit. The t±' polypeptide was purified by conventional biochemical techniques
and its potential to modulate the activity of the LDL-receptor was evaluated in a human hepatoma cell line (Hep G2) by monitoring the uptake and
degradation of labeled LDL. The LDL uptake (+ 192%) and degradation (+ 143%) by cells tested at the highest t±' dose (8 µM) were similar to
those found in cells incubated with 1 µM simvastatin, a potent inhibitor of cholesterol biosynthesis. The cell response to t±' was found to be dosedependent.
Inventor
• MORAZZONI PAOLO (IT)
• RIVA ANTONELLA (IT)
• PONZONE CESARE (IT)
• BERLANDA DAVIDE (IT)
• DURANTI MARCELLO (IT)
• CONSONNI ALESSANDRO (IT)
Applicant
INDENA SPA (IT)
IPC 8 full level (invention and additional information)
C07K 14/415 (2006.01)
CPC (invention and additional information)
C07K 14/415 (2013.01)
Citation (applicant)
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
SIRTORI CR ET AL. CURR ATHEROSCLER REP. vol. 3, 2001, pages 47 - 53
LOVATI MR ET AL. J NUTR. vol. 122, 1992, pages 1971 - 8
MANZONI C ET AL. J NUTR. vol. 133, 2003, pages 2149 - 55
DURANTI M. ET AL. J NUTR. vol. 134, 2004, pages 1334 - 39
THANH, VH ET AL. BIOCHIM BIOPHYS ACTA vol. 439, 1976, pages 326 - 38
MARUYAMA N ET AL. J AGRIC FOOD CHEM. vol. 47, 1999, pages 5278 - 84
SIRTORI CR ET AL. NUTR METAB CARDIOVASC DIS. vol. 8, 1998, pages 334 - 40
LOVATI MR ET AL. J AGRIC FOOD CHEM. vol. 46, 1998, pages 2474 - 80
LOVATI MR ET AL. J NUTR. vol. 130, 2000, pages 2543 - 2549
LOVATI MR ET AL. FASEB J. vol. LB 391, 2006, page 86
LAEMMLI UK NATURE vol. 227, 1970, pages 660 - 5
HAVEL RY ET AL. J CLIN INVEST. vol. 34, 1955, pages 1345 - 53
BILHEIMER DW ET AL. BIOCHIM. BIOPHYS. ACTA vol. 260, 1972, pages 212 - 21
SIRTORI CR ET AL. BRIT J NUTR. vol. 97, 2007, pages 816 - 22
LOVATI MR ET AL. NUTR METAB CARDIOVASC DIS. vol. 1, 1991, pages 18 - 24
LOVATI MR ET AL. J CLIN INVEST. vol. 80, 1987, pages 1498 - 502
KITTS DD ET AL. CURR PHARM DES. vol. 9, 2003, pages 1309 - 23
WAN XS ET AL. NUTRITION AND CANCER vol. 43, 2002, pages 167 - 73
CLEMENTE A ET AL. RECENT PROGRESS IN MEDICINAL PLANTS vol. 20, 2008, pages 397 - 417
MATOBA N ET AL. FEBS LETT. vol. 497, 2001, pages 50 - 4
CHO SJ ET AL. J AGRIC FOOD CHEM. vol. 56, 2008, pages 4372 - 6
CASTIGLIONI S ET AL. ATHEROSCLEROSIS vol. 171, 2003, pages 163 - 70
SIRTORI CR ET AL. DRUG SAFETY vol. 24, 2001, pages 665 - 82
Citation (search report)
• [X] WO 03063608 A1
• [X] WO 2007147064 A2
• [X] M LOVATI: "Biological activity of a truncated form of the soybean 7S globulin alpha subunit on cell
cholesterol homeostasis" ATHEROSCLEROSIS SUPPLEMENTS 2009, [Online] vol. 10, no. 2, 16 June 2009
(2009-06-16), XP002551995 Retrieved from the Internet: URL:http://ex2.excerptamedica.com/09isa/index.cfm?
fuseaction=CIS2002&hoofdnav=Search&content=zk.results_all&topicselected=*&searchtext=lovati&what=AUTHOR&selection=ALL&abstrn
[retrieved on 2009-10-22] -& "XV International Symposium on Atherosclerosis, June 14-18, 2009: Timetable for Tuesday,
16.6.2009"[Online] 16 June 2009 (2009-06-16), pages 75-94, XP002551996 Retrieved from the Internet: URL:http://
www.isa2009.org/download/Tuesday.pdf> [retrieved on 2009-10-22]
• [A] DURANTI MARCELLO ET AL: "The alpha' subunit from soybean 7S globulin lowers plasma lipids and upregulates liver betaVLDL receptors in rat feed a hypercholesterolemic diet" JOURNAL OF NUTRITION, vol. 134, no. 6, June 2004 (2004-06), pages
1334-1339, XP002551997 ISSN: 0022-3166
Designated contracting state (EPC)
AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO SE SI SK TR
Designated extension state (EPC)
AL BA RS
EPO simple patent family
EP 2264054 A1 20101222; WO 2010145819 A1 20101223
INPADOC legal status
2012-05-02 [18D] DEEMED TO BE WITHDRAWN
- Effective date : 20111103
2011-07-20 [17Q] FIRST EXAMINATION REPORT
- Effective date : 20110621
2011-07-06 [17P] REQUEST FOR EXAMINATION FILED
- Effective date : 20110530
2010-12-22 [AK] DESIGNATED CONTRACTING STATES:
- Kind Code of Ref Document : A1
- Designated State(s) : AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT
RO SE SI SK TR
2010-12-22 [AX] REQUEST FOR EXTENSION OF THE EUROPEAN PATENT TO
- Countries concerned : AL BA RS