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Method of modulating b cell functioning

A B cell, functional technology, applied in extracellular fluid diseases, antibacterial drugs, pharmaceutical formulations, etc., can solve problems such as death, activation, infection, etc.

Inactive Publication Date: 2008-01-02
太平洋治疗学控股有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Disease course is variable but often fatal in the actively progressive form due to infection or complications of treatment
The cause of the disease is unknown, but Epstein-Barr virus (EBV) infection is known to cause activation of synovial fluid B lymphocytes to produce abnormal IgG antibodies

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0229] Efficacy of tranilast on collagen-induced arthritis that has occurred

[0230] Materials and methods

[0231] Preparation of Type II Collagen

[0232] Bovine type II collagen (CII) was purified and prepared as previously described (9) and stirred overnight at 4°C in 0.1 M acetic acid to dissolve it.

[0233] Immunized mice

[0234] Bovine CII (200 μg / mouse) emulsified with Complete Freund's Adjuvant (CFA) (Difco Laboratories, West Moseley, UK) for 8-12 week-old male DBA / 1 mice (7-8 animals / group) Intradermal (i.d.) immunizations were performed. Mice were checked daily for signs of arthritis starting on day 14 after immunization, and treatment was initiated on day 1 of the onset of arthritis. The study was approved by the local Ethical Review Process Committee and the Home Office of the United Kingdom.

[0235] treat arthritis

[0236] Dissolve Tranilast in 1% NaHCO by heating to 70 °C 3 , and injected intraperitoneally (i.p.) at 100, 200, or 400 mg / kg / day.

[02...

Embodiment 2

[0247] Detection of B cell proliferation by FACS analysis

[0248] Materials and methods

[0249] B cell purification

[0250] All centrifugations were performed at 1500 rpm for 5 minutes.

[0251] B cells were prepared from mouse spleens using rabbit anti-mouse IgM beads and the MACS system.

[0252] Remove 3 spleens from 8-12 week old male DBA / 1 mice. Prepare a single cell suspension by passing it through a cell sieve.

[0253] Red blood cells were lysed by adding 5 ml of red blood cell lysis buffer (Sigma) and incubating for 5 minutes. Add 5ml RPMI to the cell suspension, wash twice, and count live cells by trypan blue (Sigma) staining.

[0254] Take 90μl / 10×10 6 Cells were resuspended in IMAG buffer (BD) at 10 x 10 6 Cells were spiked with 10 μl of rat anti-mouse IgM microbeads (MACS).

[0255] The cell-bead suspension was incubated in the refrigerator for 15 minutes.

[0256] Place mini MACS columns (each 7 x 10 7 Cells (1 column) were placed on a magnet (MACS) ...

Embodiment 3

[0285] Proliferating B cells [ 3 H] Detection

[0286] Materials and methods

[0287] Purification and stimulation of B cells - as described for FACS.

[0288] All centrifugations were performed at 1500 rpm for 5 minutes.

[0289] B cells were prepared from mouse spleens using rabbit anti-mouse IgM beads and the MACS system.

[0290] Remove 3 spleens from 8-12 week old male DBA / 1 mice. Prepare a single cell suspension by passing it through a cell sieve.

[0291] Red blood cells were lysed by adding 5 ml of red blood cell lysis buffer (Sigma) and incubating for 5 minutes. Add 5ml RPMI to the cell suspension, wash twice, and count live cells by trypan blue (Sigma) staining.

[0292] Take 90μl / 10×10 6 Cells were resuspended in IMAG buffer (BD) at 10 x 10 6 Cells were spiked with 10 μl of rat anti-mouse IgM microbeads (MACS).

[0293] The cell-bead suspension was incubated in the refrigerator for 15 minutes.

[0294] Insert the mini MACS column (7 x 10 7 Cells per colum...

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PUM

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Abstract

The present invention relates generally to a method of modulating cellular functioning. More particularly, the present invention relates to a method of modulating B cell functioning, for example B cell proliferation, utilising an IDO- mediated tryptophan metabolite as herein defined (particular examples of such IDO-mediated tryptophan metabolites include 3-hydroxykynurenic acid, 3-hydroxyanthranilic acid, picolinic acid, quinolinic acid and tranilast). The method of the present invention is useful, inter alia, in the treatment and / or prophylaxis of conditions characterised by aberrant, unwanted or otherwise inappropriate B cell functioning such as antibody production, autoimmune conditions and B cell proliferation and neoplasias. In a related aspect, the present invention is directed to a method of therapeutically and / or prophylactically treating rheumatoid arthritis via the administration of the above-mentioned compounds.

Description

field of invention [0001] The present invention generally relates to methods of modulating cellular function and agents therefor. More specifically, the present invention relates to methods of modulating B cell function, such as B cell proliferation, using compounds of formula (I). In addition, the methods of the invention can be used to treat and / or prevent diseases characterized by aberrant, unwanted or otherwise inappropriate B cell function, such as autoimmune diseases and B cell neoplasia. In a related aspect, the invention relates to methods of therapeutic and / or prophylactic treatment of rheumatoid arthritis by administering a compound of formula (I). Background of the invention [0002] Bibliographic details for publications referenced by the authors in this specification are included in alphabetical order at the end of the specification. [0003] Reference in this specification to any prior art is not and should not be taken as an acknowledgment or in any way to s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/167A61K31/44A61K31/4402A61K31/47A61P3/10A61P7/06A61P17/06A61P25/02A61P29/00A61P35/00A61P35/02A61P37/06
CPCA61K31/47A61K31/4402A61K31/196A61K31/198A61K31/44A61P1/04A61P17/02A61P17/06A61P19/02A61P21/00A61P21/04A61P25/00A61P25/02A61P29/00A61P3/10A61P31/04A61P35/00A61P35/02A61P37/00A61P37/06A61P43/00A61P5/14A61P5/48A61P5/50A61P7/00A61P7/04A61P7/06
Inventor M·L·塞利J·J·英利斯R·O·威廉姆斯
Owner 太平洋治疗学控股有限公司
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