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Antibodies against human CSF-1R for use in inducing lymphocytosis in lymphomas or leukemias

A technology for lymphocytosis and leukemia cells, applied in the field of antibodies to human CSF-1R and antibodies combined with CSF-1R

Active Publication Date: 2018-03-27
F HOFFMANN LA ROCHE & CO AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CD20 is expressed on greater than 90% of B-cell non-Hodgkin's lymphomas (NHL) (Anderson, K.C. et al., Blood 63 (1984) 1424-1433), but is expressed on hematopoietic stem cells, pro-B cells, normal plasma cells, or not found in other normal tissues (Tedder, T.F. et al., J. Immunol. 135 (1985) 973-979)

Method used

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  • Antibodies against human CSF-1R for use in inducing lymphocytosis in lymphomas or leukemias
  • Antibodies against human CSF-1R for use in inducing lymphocytosis in lymphomas or leukemias
  • Antibodies against human CSF-1R for use in inducing lymphocytosis in lymphomas or leukemias

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0707] Inhibition of CSF-1-induced phosphorylation of CSF-1R in NIH3T3-CSF-1R recombinant cells

[0708] will be 4.5x10 3 In DMEM (PAA, catalog number E15-011), 2mM L-glutamine (Sigma, catalog number G7513), 2mM acetone Sodium NaCl, 1× non-essential amino acids, 10% FKS (PAA, Cat. No. A15-649) and 100 μg / ml PenStrep (Sigma, Cat. No. P4333 [10 mg / ml]) until they reached confluency. Afterwards, it was supplemented with sodium selenite [5 ng / ml] (Sigma, catalog number S9133), transferrin [10 μg / ml] (Sigma, catalog number T8158), BSA [400 μg / ml] (Roche Diagnostics GmbH, Cat. No. 10735078), 4 mM L-Glutamine (Sigma, Cat. No. G7513), 2 mM Sodium Pyruvate (Gibco, Cat. No. 11360), 1x Non-Essential Amino Acids (Gibco, Cat. No. 11140-035), 2 -Mercaptoethanol [0.05mM] (Merck, catalog number M7522), 100 μg / ml and PenStrep (Sigma, catalog number P4333) serum-free DMEM medium (PAA, catalog number E15-011) washes the cells and dissolves them in 30 μl Incubate for 16 hours in the same ...

Embodiment 2

[0714] Growth Inhibition of NIH3T3-CSF-1R Recombinant Cells in 3D Culture under Treatment with Anti-CSF-1R Monoclonal Antibody (CellTiterGlo Assay)

[0715] NIH 3T3 cells infected with retrovirus with full-length wild-type CSF-1R (SEQ ID NO: 62) or mutant CSF-1R L301S Y969F (SEQ ID NO: 63) expression vector in poly-HEMA (polymethyl Acrylate-2-hydroxyethyl ester, poly(2-hydroxyethylmethacrylate)) (Polysciences, Warrington, PA, USA)) is cultivated on the dish of coating (to prevent sticking to plastic surface) in being supplemented with 2mM L-glutamine, 2 mM sodium pyruvate and non-essential amino acids and 10% fetal calf serum (Sigma, Taufkirchen, Germany) in DMEM high glucose medium (PAA, Pasching, Austria). Cells were seeded in media replaced with serum at 5 ng / ml sodium selenite, 10 mg / ml transferrin, 400 μg / ml BSA and 0.05 mM 2-mercaptoethanol. Cells expressing wtCSF-1R form three-dimensional growth when treated with 100 ng / ml huCSF-1 (active 149 aa fragment of human C...

Embodiment 3

[0721] Inhibition of Human Macrophage Differentiation under Treatment with Anti-CSF-1R Monoclonal Antibody (CellTiterGlo Assay)

[0722] Using RosetteSep TM Human Monocytes Enrichment Cocktail (Stemcell Tech. - Cat. No. 15028) was used to isolate human monocytes from peripheral blood. Incubate the enriched monocyte population at 37 °C and 5% CO 2 seeded in 96-well microtiter plates (2.5x10 4 cells / well) in 100 μl RPMI 1640 (Gibco - catalog number 31870). When 150 ng / ml huCSF-1 was added to the medium, a clear differentiation into adherent macrophages was observed. This differentiation can be inhibited by adding anti-CSF-1R antibody. In addition, the survival of monocytes is affected and can be analyzed by CellTiterGlo (CTG) assay. Calculation of IC from concentration-dependent inhibition of monocyte survival by antibody treatment 50 (see table below).

[0723] table 5

[0724] CSF-1R Mab

[0725] In a separate test, a series of humanized forms o...

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PUM

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Abstract

The present invention relates to anti-CSF-1R induced lymphocytosis in lymphomas and leukemias and a combination therapy of antibodies which bind human CSF-1R with antibodies which bind human CD20.

Description

[0001] The present invention relates to antibodies to human CSF-1R useful in inducing lymphocytosis in lymphoma or leukemia. The present invention further relates to combination therapy of specific human CSF-1R-binding antibodies with specific human CD20-binding antibodies and anti-CD20 in combination with CSF-1R inhibitors for preventing escape from CD20-targeted therapies by targeting macrophages Methods of Antibody Targeting CD20 Expressing Cancers. Background of the invention [0002] CSF-1R and CSF-1R Antibody [0003] The human CSF-1 receptor has been known since 1986 (CSF-1R; colony-stimulating factor 1 receptor; synonyms: M-CSF receptor; macrophage colony-stimulating factor 1 receptor, Fms proto-oncogene, c-fms , SEQ ID NO:62) (Coussens, L. et al., Nature 320 (1986) 277-280). CSF-IR is a growth factor and is encoded by the c-fms proto-oncogene (for review see eg Roth, P. and Stanley, E.R., Curr. Top. Microbiol. Immunol. 181 (1992) 141-167). [0004] CSF-1R is the r...

Claims

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Application Information

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IPC IPC(8): A61K39/395
CPCC07K16/2887A61K2039/505A61K2039/507C07K2317/24C07K2317/73C07K2317/76C07K2317/90C07K16/2866A61P35/02
Inventor M·T·S·贝尔蒂拉奇欧F·卡利加里斯-卡皮奥G·加莱蒂C·克莱因C·里斯
Owner F HOFFMANN LA ROCHE & CO AG
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