US 2008.0095719A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2008/0095719 A1 Herrmann et al. (43) Pub. Date: Apr. 24, 2008 (54) BLACKBERRY EXTRACT Related U.S. Application Data (75) Inventors: Martina Herrmann, Hameln (DE); (60) Provisional application No. 60/581,307, filed on Jun. Holger Joppe, Dassel (DE); Helge 18, 2004. Provisional application No. 60/654,380, Franke, Dieburg (DE); Gabriele filed on Feb. 18, 2005. Vielhaber, Holzminden (DE) Publication Classification Correspondence Address: ROYLANCE, ABRAMS, BERDO & (51) Int. Cl. GOODMAN, L.L.P. A6IR 36/73 (2006.01) 1300 19TH STREET, N.W. 46R 8/97 (2006.01) SUTE 6OO A61O II/00 (2006.01) WASHINGTON, DC 20036 (US) (52) U.S. Cl. .............................. 424/48; 424/456; 424/58: 424/59: 424/74; 424/765 (73) Assignee: SYMRISE GMBH & CO. KG, Holz minden (DE) (57) ABSTRACT (21) Appl. No.: 11/629,753 (22) PCT Filed: Jun. 16, 2005 The invention concerns a blackberry leaf extract and its uses, in particular to slow down skin ageing, to treat the oral and (86). PCT No.: PCT/EP05/52.793 pharyngeal cavity, and there in particular to prevent and slow down periodontitis and the excessive degradation of S 371(c)(1), the periodontal connective tissue as well as damage to the (2), (4) Date: May 30, 2007 teeth caused by matrix metalloproteinases Patent Application Publication Apr. 24, 2008 US 2008/0095719 A1 FIG.1 US 2008/0095 719 A1 Apr. 24, 2008 BLACKBERRY EXTRACT higher level of MMPs was detected in light-aged skin as compared with aged skin protected from the light (J. H. 0001. The invention concerns the field of plant extracts Chung et al., J. Invest. Dermatol, 2001, 117, 1218-1224). and their uses, in particular for oral hygiene, cosmetic and The induction of matrix metalloproteinases was detected pharmaceutical purposes. The invention especially concerns both for UVA and UVB and for infrared radiation. This the production of blackberry leaf extracts, the extracts induction was able to be observed both in vitro on cultivated themselves and their use to inhibit matrix metalloproteinases human dermal fibroblasts and in vivo on UV-irradiated (MMPs), in particular to slow down skin ageing, to treat the oral and pharyngeal cavity, and there in particular to prevent human skin. Stimulation with tobacco smoke also led in and slow down periodontitis and the excessive degradation human dermal fibroblasts to an induction of the expression of periodontal connective tissue and damage to the teeth of MMP-1 and -3 (J. Krutmann, Hautarzt 2003, 54, 809 caused by matrix metalloproteinases. 817). 0007. The regulation of MMP activity can occur at three 0002 The ageing of human skin is accompanied by levels: at a transcription level, in the conversion of pro-MMP increasing formation of wrinkles and reducing elasticity and to the active form or by inactivation of MMPs by inhibitors. strength. A distinction is made in the ageing process between intrinsic and extrinsic skin ageing. Intrinsic ageing includes 0008. The reproduction of the imbalance in the pro natural changes to the skin, which are regulated by genetic teolytic activity of MMPs, in particular MMP-1, -2, -3 and makeup. The term extrinsic skin ageing stands for premature -9, brought about by intrinsic and extrinsic skin ageing is ageing processes brought about by exogenous influences thus an important objective in the development of new Such as Sunlight, environmental pollutants (for example cosmetic active ingredients against skin ageing and oZone, tobacco smoke, etc.), psychological stress and wrinkles. chronic inflammation. Ultraviolet radiation is the most 0009. The use of MMP-1-inhibiting substances (retinyl important exogenous pathogen leading to premature ageing palmitate, propyl gallate, precocene, 6-hydroxy-7-methoxy of the human skin (light ageing). In addition to natural 2,2-dimethyl-1(2H)-benzopyran, 3,4-dihydro-6-hydroxy-7- sunlight, irradiation of the human skin with artificial UV methoxy-2,2-dimethyl-1 (2H)-benzopyran) to prevent Sun radiation (Solarium) is playing an ever greater role. light- and/or heat-induced ageing of the human skin was 0003. The structural changes responsible for the clinical described in the laid-open specification WO 01/74320. image of aged skin take place primarily in the dermis. 0010 Matrix metalloproteinases are also significant in Elasticity and strength of the skin are determined substan pathological changes to the periodontium. Periodontitis is an tially by the two main constituents of the dermal extracel inflammation of the periodontium, in other words the tissues lular matrix, the two fibre proteins collagen and elastin. The that surround and support the teeth. The periodontium dermis contains mainly collagen-1 (90-85%), which is comprises various tissues: the gum epithelium (gingiva), the formed exclusively from dermal fibroblasts, and, in signifi connective tissue of the gingiva, the periodontal ligament cantly smaller amounts (10-15%), collagen-3. Elastin, the (desmodontium), the cementum and the Surrounding alveo principal component of the elastic fibres of the skin apart lar bone. The desmodontium is located between the surface from fibrillin, is contained in the dermis in a proportion of of the root and the alveolar bone. It is a cell-rich connective around 1-3%. tissue which holds the tooth in the bony tooth socket, the 0004. In comparison to young skin, old skin is charac alveolus. 53 to 74% of the periodontal space is made up of terised by a lowering concentration of collagen and elastin. collagen and oxytalan fibre bundles. The portion of the This age-related loss of tissue is at least partially caused by periodontal fibres incorporated into the cementum and the an imbalance between activation and inhibition of pro alveolar bone holds the tooth in the alveolus. The main teolytic activity. An important role is played here by matrix clinical features of periodontitis include inflammation of the metalloproteinases, a group of enzymes which are able to gums, attachment loss, formation of periodontal pockets and break down the macromolecules in the extracellular matrix degradation of the alveolar bone. (ECM) proteolytically. Thus it has been found that the 0011. The main cause of periodontitis is plaque. This content of MMPs is markedly higher in old skin than in consists of certain components of saliva, food residues and young skin (J. H. Chung et al., J. Invest. Dermatol, 2001, above all bacteria and their decomposition products. This 117, 1218-1224). special form of an infectious disease is caused in most cases 0005. MMPs have a broad, often overlapping, substrate by Porphyromonas gingivalis, Bacteroides forsythus and specificity, and when combined they are capable of destroy Actinobacillus actinomycetemcomitans. The continuous ing all protein components of the extracellular matrix. Some release of bacterial toxins, especially of lipopolysaccharides, 20 MMPs have been identified to date. They are generally presumably triggers the distribution of proinflammatory secreted as inactive pro-enzymes (pro-MMP). Of particular mediators, such as IL-1beta, TNF-alpha and PGE2 for importance in the human skin are, above all, MMP-1 (col example, in the patients affected tissues. These signal lagenase-1), MMP-2 (gelatinase A), MMP-9 (gelatinase B) Substances stimulate the infiltration of immunocompetent and MMP-3. In addition to collagen-1 and -3, MMP-1 also cells into the populated tissue. The migration of neutrophilic cleaves pro-MMP-2 and pro-MMP-9, thereby causing them granulocytes and macrophages then Subsequently leads to to be activated. MMP-2 and MMP-9 belong to the elastin inflammation of the gums (gingivitis) and to the release of degrading proteases (A. Thibodeau, Cosmetics & Toiletries proinflammatory mediators such as IL-1 and IL-6, for example. These in turn activate in the skin and mucous 2000, 115 (11), 75-82). membranes the synthesis of matrix-degrading metallopro 0006 MMPs also play a critical role in the premature teinases (matrix metalloproteinases, MMPs), which destroy skin ageing caused by exogenous factors. Thus, an even the extracellular matrix of the Surrounding connective tissue. US 2008/0095 719 A1 Apr. 24, 2008 This allows bacteria, which initially interacted with the free genase-1), MMP-2 (gelatinase A), MMP-8 (collagenase-2) gingiva, to penetrate further into the underlying connective and MMP-9 (gelatinase B) must be inhibited at a very early tissue, continuing inflammation processes and the synthesis Stage. of MMPs there and finally loosening the connection between the uppermost layer of the epithelium and the root of the 0017. The use of synthetic MMP inhibitors in periodontal tooth. A gingival pocket is formed as a consequence. The diseases has been described in several publications (M. E. reaction of the body is the inflammation of the gingiva and Ryan et al., Curr. Opin. Periodontal. 1996, 3, 85-96; R. the periodontium with damage to the alveolar bone. In the Gendron et al., Clin. Diagn. Lab. Immunol. 1999, 6, 437 final stage of periodontitis the affected person is at risk of a 439). massive loss of teeth. 0018) A number of plant extracts have also already been described as inhibitors of various MMPs. For instance, 0012 Studies (T. Kuboto et al., Arch. Oral. Biol. 1996, numerous plant extracts, and the inhibition of various pro 41, 253-262: A. L. Ejeil et al., J. Periodontol. 2003, 74, teases, including several MMPs, that can be achieved with 188-195) have shown that the levels of a series of matrix them, are described in laid-open specification WO metalloproteinases (MMP-1, -3, -8, -9 and -13) are signifi 02/069992 A1, although no conclusions can be drawn as to cantly higher in patients with inflammation of the gums than the concentrations of extract used and the precise extraction in patients with healthy gums. The level correlates with the conditions.