University of Michigan

Solohill Engineering, Inc.

James Varani

John J Voorhees

Gary J Fisher

Sewon Kang

William J Hillegas

Hollis Showalter

David E Solomon

David L Melmoth

Andy D White

Kent J Johnson

David L Helmreich

This invention is in the field of medicinal chemistry. In particular, the invention relates to a new class of small-molecules having a dimethyl-nonatetraenyl-trimethyl-cyclohexyl structure useful as therapeutics for the treatment of subjects suffering from disorders characterized by abnormal proliferation and/or abnormal differentiation of cells, in particular of cells of which the growth and differentiation is sensitive to the actions of retinoids.

Dimethyl-nonatetraenyl-trimethyl-cyclohexyl compounds and uses thereof

Chronological aging of human skin can be delayed with the topical application of an MMP inhibitor, preferably a retinoid (an indirect MMP inhbitor); retinoids also normalize procollagen biosynthesis. Chronological aging, or natural aging, is evidenced in elderly (80+ years old) skin by increased MMP levels and decreased procollagen levels when compared with younger individuals. Prophylactic treatment of not yet chronologically-aged skin with a retinoid both inhibits degradation of dermal collagen and restores procollagen synthesis. Biopsied sections from elderly skin show that a single treatment of chronologically-aged skin with a retinoid can increase epidermal thickness, improve the dermal collagen density, and promote the formation of rete pegs and dermal papillae. Such benefits are helpful in preventing bruising, tearing, and ulceration of elderly skin. Accordingly, prophylactic treatment begun much earlier in life with an MMP inhibitor and/or a retinoid delays the onset of such symptoms.

Methods and compositions for reducing collagen loss due to chronological aging in human skin

The deleterious effects of the passage of time on human skin (i.e., chronological aging of human skin) can be prevented and treated with the topical application of a retinoid, preferably retinol. We have found that some of the same pathways (namely the stress-activated pathways, SAPs) activated in photoaging of human skin (i.e., sun-induced premature skin aging) are similarly elevated in the skin of elderly people. We have also found that other pathways (namely the mitogen-activated ERK pathway) is depressed in the same skin. Treatment of chronologically-aged skin with a non-retinoid MMP inhibitor and optionally a retinoid both inhibits degradation of dermal collagen and promotes procollagen synthesis. Biopsied sections from skin of elderly (80&plus; years old) show that a single treatment can increase epidermal thickness, improve the dermal collagen density, and promote the formation of rete pegs and dermal papillae (see FIG. ), and can decrease the amount of c-Jun and increase the amounts of Types I and III procollagen (see FIG. ). Such benefits are also helpful in preventing bruising, tearing, and ulceration of elderly skin.

Methods and compositions for preventing and treating chronological aging in human skin

A microcarrier bead system for culturing anchorage-dependent cells is formed of a polystyrene core with a coating of collagen fixed thereover. In certain embodiments, the coating is a protein, such as laminin or fibronectin. The microcarrier bead is of low density, illustratively 1.02 g/cc, and therefore requires less agitation of the nutrient media to maintain suspension. This reduced stirring causes lower shear forces to impinge upon the cells, thereby improving the attachment and proliferation of the cells being cultured. The microcarrier bead of the present invention exhibits surprising advantages with respect to cell attachment and harvesting over beads formed entirely of collagen, or of DEAE-dextran coated with collagen. During harvesting, contamination of the product resulting from dissolved collagen, particularly when proteolytic enzymes are used, is minimized. Additionally, adsorption of toxins and product by the subject microcarrier beads is minimized.

Collagen-coated polystyrene microcarrier beads

An insert for a roller bottle of the type used in cell culture for vaccine manufacture is formed of a polymeric resilient material whereby it can be bent or rolled so that it can pass through the opening of the roller bottle. Once the insert is inside the bottle, it is released so that its resilient characteristic causes it to unbend or unroll. The insert is wider than the diameter of the roller bottle, and therefore it is urged against the interior wall of the roller bottle. Apertures and/or notches are provided in the insert to increase the turbulence of a fluid and to decrease the fluidic shear forces in the roller bottle as it is rotated in conventional operation. A significant increase in productivity is achieved because particles, which may be microcarrier beads, are maintained in suspension, and prevented from sinking to the bottom of the fluid medium where cells cannot grow. The roller bottle system is thereby transformed into a microcarrier suspension culture system.

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Ann Arbor, MI, United States of America

James Varani