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(12) Patent Application Publication (10) Pub. No.: US 2003/0125264 A1 Malik (43) Pub US 2003.0125264A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2003/0125264 A1 Malik (43) Pub. Date: Jul. 3, 2003 (54) METHODS FOR TREATING WOUNDS Publication Classification (75) Inventor: Sohail Malik, Roswell, GA (US) (51) Int. CI.7. ... A61K 31/7048; A61K 31/353 (52) U.S. Cl. .............................................. 514/27; 514/456 Correspondence Address: KIMBERLY-CLARK WORLDWIDE, INC. (57) ABSTRACT 401 NORTH LAKE STREET NEENAH, WI 54956 Pharmaceutical compositions, and methods of using the Same, are provided utilizing effective amounts of one or more flavones, flavonols, flavanones, isoflavanones and (73) Assignee: Kimberly-Clark Worldwide, Inc. isoflavones to increase cell proliferation in various tissues and cell lines. AS examples, the composition and methods of (21) Appl. No.: 10/034,150 the present invention can be used to increase proliferation of fibroblast cells and, more particularly, in the treatment of (22) Filed: Dec. 29, 2001 wounds as well as Strengthening of the skin. US 2003/O125264 A1 Jul. 3, 2003 METHODS FOR TREATING WOUNDS 0005 The body produces many substances generally known as growth factorS Such as, for example, platelet FIELD OF THE INVENTION derived growth factor (PDGF), platelet-derived angiogen 0001. The present invention relates to methods and com esis factor (PDAF), vascular endothelial growth factor positions for increasing mammalian cell proliferation. (VEGF), platelet-derived epidermal growth factor (PDEGF), platelet factor 4 (PF4), transforming growth fac BACKGROUND OF THE INVENTION tor beta. (TGF-B), transforming growth factor a (TGF-A), insulin-like growth factors 1 and 2 (IGF-1 and IGF-2), beta. 0002 Damage or injury to tissues and/or organs is a thromboglobulin-related proteins (BTG), thrombospondin common occurrence. The body is most often able to isolate (TSP), fibronectin, von Wallinbrand’s factor (vWF), angio the damaged area and then repair itself by removing and genin, keratinocyte growth factor (KGF), epidermal growth replacing damaged tissue. Injury to tissues and organs can factor (EGF), fibroblast growth factor (FGF), and so forth. originate from a great variety of Sources Such as, for One of the important characteristics common to each Sub example, trauma, UV degradation, toxic and/or pathogenic stance is that each Such Substance is known or believed to degradation, thermal degradation (e.g. excessive heat or enhance cell or tissue growth. Other cell proliferating agents cold) and so forth. While the body has an impressive array have also been reported Such as, for example, those of response mechanisms that limit tissue damage and pro described in EP 0953354, JP 09-227413, JP 53-062815 A2 mote repair, methods of increasing the Speed and degree of and EP 0560845 A1. However, there exists a continuing repair are continually being Sought out. In this regard, need for additional and/or improved cell proliferating agents increasing the Speed and/or degree that injuries are healed is and compounds. In particular, there is a continuing need for beneficial in that it (i) decreases the pain and discomfort compounds that promote controlled cell growth without commonly associated with the wound and wound healing promoting or inducing deleterious or uncontrolled cell process; (ii) decreases the chances of developing an infec growth Such as, for example, cancerous growth. tion or other ailment during a period when the tissue or organ has a reduced capacity to ward of illnesses; and (iii) reduce 0006 Flavonoids are active molecules having a variety of health costs associated with treating Such conditions. important functions within the plants they are found. By way of example, certain flavonoids are believed to act as anti 0003. In this regard, and by way of example, the skin is fungal agents, anti-bacterial agents, UV-B protecting agents, the largest organ in the body and, not Surprisingly, wounds and so forth. In addition, certain flavonoids have been found and injuries to the skin are a common occurrence. Healing to have various medicinal applications Such as, for example of wounds in the skin has three general phases including (1) anti-virals, anti-inflammatories, antihistamines, anti-cancers inflammation, migration and proliferation; (2) repair which and anti-oxidants. These and other uses or functions are includes the formation of collagen and other compounds; (3) described in the following books and articles: B. Bohm, wound closure. Initially, inflammatory cells and other cells Introduction To Flavonoids Chapters 7-8 (1998); J. Har migrate into and fill the damaged area. Then, in the repair bourne et al., Advances. In Flavonoid Research Since 1992, phase, new connective tissues are formed from fibronectin, 55 Phytochemistry 481-500 (2000); A. L. Miller, Antioxi which in turn results in the production of collagen fibrils and dant Flavonoids. Structure, Function and Clinical Usage, eventually larger collagen fibers. The wound is thereafter Alternate Medicine Review, 1, 103-111 (1996); R. Nijveldt closed by wound contraction which results, in part, by the et al., Flavonoids: A Review Of Probable Mechanisms Of modified fibroblasts present in and around the wound. Action And Potential Applications, American Journal Of 0004 Fibroblasts, endothelial cells and keratinocytes are Clinical Nutrition, 74, 418-425 (2001). Previous studies indispensable in cutaneous wound repair. All three cell types have focused significantly on the inhibitory action of fla play Vital roles in the initial phase of wound healing. vonoids against enzyme Systems, inflammation and cancer Fibroblasts migrate into the wound site within about 24 see, for example, J. Harborne, Editor; The Flavonoids. hours after injury. During a later phase of healing (typically Advances in Research Since 1986, (1994). Moreover, fla about 4-21 days), fibroblasts are activated and undergo a vones, flavonols, flavanones and isoflavones have been burst of proliferative and synthetic activity. They produce shown to possess anti-proliferative activity for numerous high amount of fibronectin and Synthesize other proteina cell lines in vitro. See, S. Kuntz et al., Comparative Analysis ceous components of extracellular matrix, including col Of The Effects Of Flavonoids. On Proliferation, Cytotoxicity lagen, elastin and glycosaminglycans. Fibroblasts are also And Apoptosis. In Human Colon Cancer Cell Lines, Euro known to contribute in contraction of the wound. Accord pean Journal Of Nutrition 38, 133-142 (1999). The authors ingly, fibroblast proliferating agents have therefore been of the aforesaid article demonstrated that this inhibition was shown to increase the wound healing process. See, for not a consequence of cytotoxic effects and concluded that example, S. Casadio et al., On the Healing Properties of these four classes of flavonoids inhibit in vitro growth by Esters of D-panthenol with Terpene Acids, with Particular cell cycle arrest. Reference to D-pantothenyl Trifarnesylacetate, Arzneimit telforschung, 17, 1122-1125, (1967); M. Aprahamian et al., SUMMARY OF INVENTION Effects of Supplemental Pantothenic Acid on Wound Heal 0007. It has been surprisingly found that flavonoids, ing. Experimental Study in Rabbit, American Journal of when used in the proper dosages and/or amounts, Signifi Clinical Nutrition 41,578-589 (1985); B. J. Weimann and et cantly increase cell proliferation and thus can be used to treat al., Studies on Wound Healing. Effects of Calcium D-Pan wounds as well as other conditions or maladies where tothenate on the Migration, Proliferation and Protein Syn increased cell growth would be beneficial. In one aspect, a thesis of Human Dermal Fibroblasts in Culture, Interna cell proliferating composition is provided comprising a tional Journal of Vitamin Nutrition Res., 69, 113-119, therapeutically effective amount of a flavonoid selected from (1999). the group consisting of flavones, flavonols, flavanones, US 2003/O125264 A1 Jul. 3, 2003 isoflavanones and isoflavones. In one embodiment, fla canines, equines, felines, etc. AS Specific examples, mam vonoid is present in the composition in an amount between mals include humans, certain farm animals (e.g. cattle, about 0.00001% and about 5% (by weight). In a specific horses, pigs, etc.), certain lab animals (e.g. mice, rats, embodiment, the flavonoid is Selected from the group con rabbits, etc.), many pets and Zoo animals (e.g. dogs, cats, Sisting of quercetin, rutin, naringin, hesperidin, Silybin, monkeys, etc.). daidzin, genistin and genistein. In a further embodiment, the 0011. The compositions and methods described herein cell proliferating composition can include one or more are believed generally Suitable for use in treating conditions pharmaceutical carriers. where cell proliferation is desirable. By way of non-limiting 0008. In an additional embodiment, a composition for example, increased proliferation of fibroblasts would be treating wounds is provided comprising (i) an effective highly beneficial in the treatment of wounds. As a further amount of a flavonoid Selected from the group consisting of example, increased proliferation of fibroblast cells would be flavones, flavonols, flavanones, isoflavanones, and isofla beneficial in the treatment of the skin by improving the vones; and (ii) a pharmaceutical carrier. In one aspect, the extra-cellular matrix thereby tightening and/or Strengthening flavonoid is present in an amount between about 0.00001% the skin. More particularly, collagen, the predominant and about 5% (by weight) of
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