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Soluble human M-CSF receptor and uses thereof

A soluble, receptor-specific technology for diagnosing disease and symptoms to address rising morbidity and mortality

Inactive Publication Date: 2014-02-05
NOVARTIS AG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Often, cancer morbidity and mortality increases significantly if it is not detected early in its development

Method used

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  • Soluble human M-CSF receptor and uses thereof
  • Soluble human M-CSF receptor and uses thereof
  • Soluble human M-CSF receptor and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] This example describes the identification of a soluble receptor for human M-CSF and shows that the soluble receptor is capable of binding M-CSF.

[0097]I. Identification of Soluble Human M-CSF Receptor in Serum Samples

[0098] A. Materials and methods

[0099] A 100 ng / ml capture antibody (DuoSet Elisa Development System hM-CSF R (R&D Systems, catalog number DY329)) working solution prepared in PBS was used to coat a microtiter plate (R&D Systems, catalog number CP0011) with 100 μl / well. Plates were sealed and incubated overnight at room temperature. After washing each well 3 times with washing buffer (0.05% Tween in PBS, pH7.2-7.4) using a multi-channel dispenser / washer, the plate was washed with 300 μl / well blocking buffer (1% BSA, 5% Sucrose, pH 7.2-7.4) for at least 1 hour at room temperature.

[0100] After repeated washing steps, samples can be added to the plate. Samples or standards diluted with diluent were added at 100 μl / well. Plates were covered with ...

Embodiment 2

[0119] This example shows that soluble M-CSF receptors are found in human urine samples. This example also shows that this receptor is present in the serum of normal subjects and breast cancer patients and that the level of soluble M-CSF receptor correlates with the level of M-CSF. Finally, this example shows that soluble M-CSF receptors are also found in primates.

[0120] I. Soluble human M-CSF receptor was also found in urine samples

[0121] Urine samples collected from healthy human volunteers were assayed for the presence of soluble human M-CSF receptor using the same ELISA setup as described in Example 1. Different levels of soluble human M-CSF receptor were detected in adult females (25-66 years), adult males (35-70 years) and children 3-9 years old. The recombinant c-fms-human Fc fusion protein was used as a reference to quantify the concentration of soluble human M-CSF receptor ( Figure 9 ).

[0122] Urine samples were collected daily throughout the menstrual cy...

Embodiment 3

[0130] This example shows that the expression of soluble human M-CSF receptor is closely related to osteoclast differentiation. This example also shows that only differentiated osteoclasts express the soluble human M-CSF receptor, the level of which increases with M-CSF removal.

[0131] A. Materials and methods

[0132] Expression of the M-CSF soluble receptor was studied in an in vitro system using human osteoclasts, in which expression of the membrane-bound M-CSF receptor was found. Specifically, the expression of soluble receptors was detected during osteoclast differentiation. Primary human osteoclast precursors (Cambrex Bio Science Walkersville, Inc., Cat. No. 2T-110) were seeded at 10,000 cells / well and 0.2ml / well in 30ng / ml human Cell culture medium with M-CSF and 100 ng / ml RANKL. Add 1 μg / mL detection antibody to each well on the same day as cell seeding. At 37°C, 5% CO 2 Cells were grown in humid air. Osteoclasts were identified by phase contrast microscopy on ...

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Abstract

Soluble human M-CSF receptor is provided, along with pharmaceutical compositions containing such receptor, kits containing a pharmaceutical composition, and methods of diagnosing and treating diseases and disorders associated with M-CSF such as bone loss in a subject afflicted with an osteolytic disease.

Description

[0001] This application is a divisional application of CN200680051912.5 filed on December 21, 2006, entitled "Soluble Human M-CSF Receptor and Its Application". technical field [0002] The present invention relates to naturally occurring soluble fragments of the human M-CSF receptor capable of binding M-CSF, methods of using the soluble fragments for therapy, and methods of diagnosing diseases and conditions by detecting the soluble fragments in patient samples. Background of the invention [0003] An estimated 1.4 million new cases of cancer are estimated to occur each year, and an estimated 600,000 people die from cancer each year. With the development of detection and treatment, many patients survive for a long time. As of January 1, 2002, there were an estimated 10.1 million cancer survivors, approximately 3.4% of all patients. Among these cancer survivors, the most common cancer sites were breast (22%), prostate (18%), colorectal (10%), and gynecologic (10%). [0004...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/574G01N33/57488G01N33/689G01N2333/535
Inventor C·刘M·多特-莱因哈德J·库尼奇M·卡瓦诺
Owner NOVARTIS AG
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