Method for upregulating antigen expression

Inactive Publication Date: 2016-04-14
NORDIC NANOVECTOR
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0030]In another embodiment the radioimmunotherapy is given as a low dose which by it

Problems solved by technology

However it was not known if low doserate radioimmunotherapy could upregulate antigen expression.
A problem with this approach is that experimental work and clinical data have indica

Method used

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  • Method for upregulating antigen expression
  • Method for upregulating antigen expression
  • Method for upregulating antigen expression

Examples

Experimental program
Comparison scheme
Effect test

example 1

Treatment with Betalutin (177Lu-tetraxetan-tetulomab, or 177Lu-HH1) and Effect on Antigen Expression

[0182]Materials & Methods:

[0183]Labelling of Antibodies

[0184]Labelling of the antibody HH-1 with p-SCN-Bn-DOTA and 177Lu was performed following standard procedures described in Dahle et al. (2013) and Repetto-Llamazares et al. (2013). The radiochemical purity (RCP) of the radioimmunoconjugate (RIC) was evaluated using Instant Thin Layer Chromatography. If RCP was below 95% the conjugate was purified using Econo-Pac 10 DG columns (Bio-rad Laboratories, California, USA). RCP was higher than 97% for all RICs used in the experiments. The specific activity of Betalutin was 92 MBq / kg with an antibody concentration of 178 μg / ml. The immunoreactive fractions (IRFs) of the RICs were estimated using Ramos and Daudi lymphoma cells and a one point assay. A cell concentration of 75 million cells / ml were used. Cells incubated with cold tetulomab were used to assess the non-specific binding. Blocke...

example 2

Introduction

[0194]To evaluate if the effect from 177Lu-tetulomab was due to cell specific radiation, Daudi cells were incubated with Betalutin for 1 h.

[0195]Materials and Methods

[0196]Labelling of Antibodies

[0197]The procedure used to label the antibodies and to calculate the IRF has been previously discussed in Example 1. The specific activity of 177Lu-tetulomab was 212 MBq / mg and the Ab concentration was 0.5 mg / ml. The IRF was 72+ / −2% and it was measured in Ramos cells.

[0198]Treatment of Cells

[0199]The procedure followed to treat the cells was similar to the one described in example 1. Two different treatments were evaluated in this example. One treatment involved incubation of cells during 24 hours, similarly to what was done in example 1. The other treatment involved incubation of cells during 1 hour. Control cells were prepared for each of the treatments as described in example 1. The same amount of activity was added to both treatments (4.6 MBq, the same activity as given in e...

example 3

Introduction

[0204]To study if the upregulation of antigens was a general feature of lymphoma rather than a cell line specific effect, Daudi cells were tested together with the three other lymphoma cells lines, Raji, Ramos and Rec-1.

[0205]Materials and Methods

[0206]Labelling of Antibodies

[0207]The procedure used to label the antibodies and to calculate the IRF has been previously discussed in Example 1. The specific activity of 177Lu-tetulomab was 212 MBq / mg and the Ab concentration was 0.5 mg / ml. The IRF was 72+ / −2% and it was measured in Ramos cells.

[0208]Treatment of Cells

[0209]The procedure followed to treat the cells was similar to the one described in example 1. Cells were incubated during 1 hour. Control cells were prepared for each of the cell lines as described in example 1. The same amount of activity was added to the cell lines (4.6 MBq, the same activity as given in example 1). The dose to the medium of the cells was around 0.05 Gy. Every 2 to 5 days cells were split and ...

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Abstract

The present invention relates to radioimmunoconjugates that are capable of upregulating expression of one or more antigens. The upregulated antigens can be the antigens that are targeted by the radioimmunoconjugates or different antigens expressed on the same cells. The present invention also relates to methods of treating cancer and diseases and disorders of the immune system by utilizing this enhanced expression of antigens.

Description

TECHNICAL FIELD OF THE INVENTION[0001]The present invention relates to radioimmunoconjugates that are capable of upregulating expression of one or more antigens. The upregulated antigens can be the antigens that are targeted by the radioimmunoconjugates or different antigens expressed on the same cells.BACKGROUND OF THE INVENTION[0002]The use of monoclonal antibody therapy (MAT) has been an area of great interest and strong research efforts. To be effective, MAT depends on a sufficient number of antigens per cell of which the antigens can bind to for triggering response (Sugimoto et al., 2009, Czuczman et al., 2008).[0003]The use of radioimmunotherapy (RIT) has been approved as a therapeutic option in cancer therapy (Stevens et al., 2012). Today radioimmunotherapy is mainly used in those patient experiencing relapse from chemotherapy and / or MAT (Stevens et al., 2012).[0004]It is known from the literature that high dose rate external beam irradiation can cause an increase of CD20 in ...

Claims

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Application Information

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IPC IPC(8): A61K51/10A61K39/395
CPCA61K51/1027A61K2039/507A61K51/1096A61K39/39558C07K16/2887C07K16/2896A61P35/00A61P35/02A61P43/00A61K2300/00A61K39/395A61K51/1093
Inventor LARSEN, ROY H.REPETTO-LLAMAZARES, ADA
Owner NORDIC NANOVECTOR
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