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Use of Insulin for the Treatment of Cartilaginous Disorders

Inactive Publication Date: 2007-03-01
GENENTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0023] In another embodiment, the invention concerns a method of maintaining, enhancing or promoting the growth of chondrocytes in serum-free culture by contacting the chondrocytes with an effective amount of insulin or insulin variant. Alternatively, the method concerns contacting the chondrocyte with an effective amount of insulin or insulin variant in a sustained or extended-release formulation. Alternatively, the present invention concerns a method of stimulating the regeneration or preventing the degradation of cartilage resulting from injury or a cartilaginous disorder by transplantation of an effective amount of chondrocytes previously treated with an effective amount of insulin or insulin variant.

Problems solved by technology

In a specific aspect, the injury can be the result of excessive mechanical stress or other biomechanical instability resulting from a sports injury or obesity.

Method used

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  • Use of Insulin for the Treatment of Cartilaginous Disorders
  • Use of Insulin for the Treatment of Cartilaginous Disorders
  • Use of Insulin for the Treatment of Cartilaginous Disorders

Examples

Experimental program
Comparison scheme
Effect test

example 1

Effect on Insulin on Primary Articular Chondrocytes

Introduction

[0325] This experiment shows the effect of various concentrations (0.1-100 nM) of insulin on matrix (proteoglycan) synthesis and on viability of chondrocytes in serum-free culture media. In order to culture chondrocytes, articular cartilage is digested with enzymes which remove the extracellular matrix. Thus, the cellular environment in this culture system may be similar to that found in later stages of cartilage disorders where the matrix has been depleted. Since essentially all of the matrix synthesized by chondrocytes cultured in monolayer is secreted into the media, the amount of proteoglycans in the media of such cells is indicative of matrix synthesis. Proteoglycans are measured in media using the 1,9-dimethylmethylene blue (DMMB) colorimetric assay of Farndale and Buttle, Biochim. Biophys. Acta 883: 173-177 (1986). In this assay, the change in color of the DMMB dye which occurs upon its binding to proteoglycans...

example 2

Articular Cartilage Explant Assay

Introduction

[0337] The experiments of this example examine both the synthetic and prophylactic potential of the test compound on cartilage matrix turnover. This potential is determined by measuring matrix (i.e proteoglycan) synthesis and breakdown, as well as nitric oxide production, in articular cartilage. These parameters are evaluated in the presence and absence of interleukin 1α, IL-1α. Articular cartilage explants have several advantages over primary cells in culture. First, and perhaps most importantly, cells in explants remain embedded in tissue architecture produced in vivo. Secondly, these explants are phenotypically stable for several weeks ex vivo, during which time they are able to maintain tissue homeostasis. Finally, unlike primary cells, explants can be used to measure matrix breakdown. To set up cartilage explants, articular cartilage must be dissected and minced which results in disruption of the collagen network and release of pr...

example 3

Mouse Patellae Assay

Introduction

[0348] This assay determines the in vitro and in vivo effect of the tested compound on proteoglycan synthesis in the patellae of mice. The patella is a very useful model to study the effects of the test compound because it permits the evaluation on cartilage which has not been removed from the underlying bone. Moreover, since each animal has one patellae in each leg, experiments can be performed using the contralateral joint as a control. This assay involves injection of a protein into the intra-articular space of a (mouse) knee joint, and subsequent harvest (within a few days after injection) of the patella (kneecap) for measurement of matrix synthesis (FIG. 6). The procedure performed herein, and outlined in FIG. 6, has been previously used to measure effects of cytokines in vitro and in vivo (Van den Berg et al., Rheum. Int. 1: 165-9 (1982); Vershure P. J. et al., Ann. Rheum. Dis. 53: 455-460 (1994); and Van de Loo et al., Arthit. Rheum. 38: 164...

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Abstract

The present invention relates to methods for the treatment and repair of cartilage, including cartilage damaged by injury or cartilaginous disorders, including arthritis, comprising the administration of insulin and / or insulin variants. Optionally, the administration may be in combination with a cartilage agent (e.g., peptide growth factor, catabolism antagonist, osteo-, synovial, anti-inflammatory factor), in an extended- or sustained-release form. Alternatively, the method provides for the treatment and repair of cartilage damaged by injury or cartilaginous disorders comprising the administration of insulin and / or insulin in combination with standard surgical techniques. Alternatively, the method provides for the treatment and repair of cartilage damaged by injury or cartilaginous disorders comprising the administration of chondrocytes previously treated with an effective amount of insulin and / or insulin variant.

Description

RELATED APPLICATIONS [0001] This application is a continuation of U.S. Ser. No. 10 / 740,098 filed Dec. 17, 2003, which is a continuation of U.S. Ser. No. 09 / 815,229 filed Mar. 22, 2001 (U.S. Pat. No. 6,689,747), which claims priority under 35 USC 119(e) to provisional application No. 60 / 192,103 filed Mar. 24, 2000; all of which are incorporated herein by reference.FIELD OF THE INVENTION [0002] The present invention relates generally to the repair of cartilage and the treatment of cartilaginous disorders. BACKGROUND OF THE INVENTION [0003] Cartilaginous disorders broadly describe a collection of diseases characterized by degeneration of or metabolic abnormalities in the connective tissues which are manifested by pain, stiffness and limitation of motion of the affected body parts. The origin of these disorders can be pathological or as a result of trauma or injury. [0004] Osteoarthritis (OA), also known as osteoarthrosis or degenerative joint disease, is the result of a series of local...

Claims

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Application Information

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IPC IPC(8): A61K38/28A61K9/14C12N15/09A61K9/52A61K38/00A61K38/27A61K38/30A61K38/55A61K39/395A61K45/00A61K47/12A61K47/34A61P19/02A61P19/08A61P29/00A61P31/00A61P37/00C07K14/62
CPCA61K9/5031A61K38/28A61K38/30A61K38/55A61K39/3955A61K2300/00A61P1/04A61P1/16A61P11/00A61P11/02A61P11/06A61P13/12A61P17/00A61P17/04A61P19/00A61P19/02A61P19/06A61P19/08A61P21/00A61P25/00A61P27/02A61P29/00A61P3/04A61P31/00A61P31/04A61P31/10A61P31/12A61P33/00A61P37/00A61P37/06A61P37/08A61P5/14A61P7/00A61P7/06A61P9/00A61P3/10
Inventor FILVAROFF, ELLEN H.OKUMU, FRANKLIN H.
Owner GENENTECH INC
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