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Alpha-lactalbumin composition

A technology of whey protein and composition, which is applied in the direction of drug combination, albumin peptide, peptide/protein composition, etc.

Inactive Publication Date: 2009-11-25
HAMLET PHARMA AB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, once the change has occurred, the presence of calcium or a functional calcium binding site does not result in any loss of activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0426]Purification and conversion of bovine α-lactalbumin into bLAC

[0427] Whole milk (2 L) was skimmed by centrifugation at 3500 xg for 30 minutes. Skim milk was subjected to ammonium sulfate precipitation overnight (264 g / L = 40-45% saturation), after which the precipitate was centrifuged at 3500 xg for 30 minutes. The ammonium sulfate precipitated supernatant was harvested and filtered first through a paper filter to remove any remaining precipitate or fat, then through a Millipore Optiscale filter with Polysep II media 1.0 / 0.5 μm.

[0428] The ammonium sulfate precipitated supernatant was made suitable for HIC chromatography by adding 32.55 mL EDTA (0.25M) + 24.97 mL Tris-EDTA (Tris 50 mM, EDTA 1 mM, pH 7.5) + 67.45 mL per 100 mL ammonium sulfate precipitated supernatant. Conditioned medium was adjusted to pH 7.5.

[0429] By filling 300mL phenyl sepharose 6FF High Sub (GE Healthcare) with a bed height of 15cm and an area of ​​19.6cm 2 Bovine α-lactalbumin was purifie...

Embodiment 2

[0449] Cytotoxicity of monomeric α-lactalbumin compositions

[0450] As outlined below and in the Figure 4 As shown in, by adopting ViaLight from Cambrex PLUS cell proliferation and cytotoxicity kit (Cell Proliferation and Cytotoxicity BioAssay Kit), test the cytotoxic activity of α-lactalbumin composition by viability test. The potency of LAC preparations is derived from their killing of the murine lymphocytic leukemia cell line L1210 (ATCC Competence Assay Cat. No. CCL-219).

[0451] For each LAC formulation tested, an appropriate serial dilution was prepared in 0.9% NaCl solution. Mix 20 μL of each dilution (in triplicate) on a 96-well white-walled cell culture plate with 50 μL of cell suspension containing 100,000 or 200,000 PBS-washed L1210 cells in RPMI 1640 medium without serum and HEPES . at 37°C and 5% CO 2 After 1 hour of incubation at 0°C, 5 μL of fetal calf serum was added to each well to inactivate all extracellular LACs. at 37°C and 5% CO 2 After an addi...

Embodiment 3

[0460] HIC loading as a determinant of monomeric / multimeric bLAC ratio in α-lactalbumin complexes

[0461] The monomer / polymer bLAC ratio can be controlled by the monomer / polymer composition of bLA purified by hydrophobic interaction chromatography as described in Example 1.

[0462] 2mg / mL gel (32mg / cm 2 ) loading resulted in bLA containing both dimeric and monomeric forms ( Figure 8 ), while 6mg / mL resin (90mg / cm 2 ) loading result is only containing monomer bLA (embodiment 1, image 3 ). Transformation of bLA was performed as described in Example 1.

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Abstract

The present invention relates to a pharmaceutical composition comprising mono- meric alpha-lactalbumin complex, preferably LAC, which is an active complex of alpha-lactalbumin and a fatty acid or lipid with selective cytotoxic activity. The composition of the invention comprises insignificant amounts of oligomeric / multi- meric alpha-lactalbumin complex, preferably LAC. Based on the selective cyto- toxicity of the alpha-lactalbumin complex, preferably LAC composition such compositions are suitable for use in the manufacture of medicaments for use in therapy. Medicaments, comprising monomeric LAC are for use in the treatment of bacterial and viral infections and in particular cancer due to the selective cytotoxic activity. The application further relates to methods of producing a composition comprising monomeric alpha-lactalbumin complex, preferably LAC with cytotoxic activity.

Description

[0001] All patent and non-patent references cited in said application or this application are also hereby incorporated by reference in their entirety. field of invention [0002] The present invention relates to pharmaceutical compositions comprising a monomeric alpha-lactalbumin complex, preferably LAC, an active complex of alpha-lactalbumin with fatty acids or lipids having selective cytotoxic activity. The composition of the invention comprises a small amount of oligomeric / polymeric alpha-lactalbumin complex, preferably LAC. Based on the selective cytotoxicity of alpha-lactalbumin complexes, preferably LAC-based compositions, such compositions are suitable for the manufacture of medicaments for therapy. Drugs comprising monomeric LAC are useful in the treatment of bacterial and viral infections and especially in the treatment of cancer due to their selective cytotoxic activity. [0003] The present application further relates to methods of producing compositions comprising...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/48A61K38/38A61K31/20C07K14/76A61P35/00A61P31/02
CPCA61K38/38A61K31/20A61K47/48046A61K47/48038A61K47/542A61K47/543A61P11/00A61P17/00A61P31/00A61P31/02A61P35/00A61K2300/00
Inventor J·西格M·古尔德曼F·马蒂森L·康格斯莱夫
Owner HAMLET PHARMA AB
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