HAS-active ingredient conjugates

a technology of active ingredients and conjugates, which is applied in the field of conjugates of active ingredients, can solve the problems of preventing the medical use of these products, affecting the clinical use of many active ingredients of pharmaceuticals, and provoking life-threatening hypersensitivity reactions,

Inactive Publication Date: 2006-08-24
FRESENIUS KABI DEUT GMBH
View PDF60 Cites 21 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0028] The object of the present invention is thus to provide improved hydroxyalkyl starch-active ingredient conjugates and processes for their preparation which lead to biologically active conjugates which can be used in everyday clinical practice. A further object of the present invention is to provide a process for the preparation of hydroxyalkyl starch-active ingredient conjugates wherein by-products are not produced in significant quantities, as these by-products also adversely affect the subsequent purification of the product to a significant extent.

Problems solved by technology

The clinical use of many active ingredients of pharmaceuticals is adversely affected by a number of problems (e.g. Delgado et al., Critical Reviews in Therapeutic Drug Carrier Systems, Vol. 9 (3, 4), (1992) pp.
Proteins expressed by bacteria as well as other recombinant proteins can have an increased immunogenicity and provoke life-threatening hypersensitivity reactions.
Corresponding reactions naturally prevent the medical use of these products.
The process of coupling the active ingredients proved to be problematic however, as the active group of the protein was inactivated by coupling to PEG or the reactions did not provide the reaction product in a suitable yield.
However, although the demand for such a product was recognized early on (e.g. Rabiner, J. Exp. Med. 126, (1967) 1127), none of the known HBOC products has as yet achieved the status of an approved drug.
Isolated haemoglobin molecules are however very unstable and rapidly break down into the more stable α,β dimers (MW 32 kDa).
Thus, the protein-HES conjugates known in the state of the art result from a non-selective coupling of Bromcyan-activated HES to haemoglobin (e.g. DE 26 16 086, hereby incorporated by reference) Corresponding processes can lead to polydisperse products with non-uniform properties and potentially toxic side effects.
In every case, a costly removal of the organic solvent is necessary, as even residual proportions of organic solvents may not be acceptable for the intended medical use.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • HAS-active ingredient conjugates
  • HAS-active ingredient conjugates
  • HAS-active ingredient conjugates

Examples

Experimental program
Comparison scheme
Effect test

example 1

Selective Oxidation of the Reducing End Groups of the Hydroxyethyl Starch

[0141] For the selective oxidation of the reducing end groups of the hydroxyethyl starch (130 kD and 10 kD), the same were dissolved in a minimal quantity of water and reacted with different quantities of an iodine solution and a KOH solution.

[0142] The mixture was stirred until the colour indicating I2 disappeared. This procedure was repeated several times in order to achieve the addition of a larger quantity of the iodine solution and KOH solution. The solution was subsequently purified using an Amberlite IR 120 Na+ cation-exchanger resin, dialysed for 20 hours against distilled water (dialysis tube with an exclusion limit of 4-6 kD) and lyopholized.

[0143] The degree of oxidation was determined in each case using the process disclosed in Somogyi, N. (Method in Carbohydride Chemistry, Vol. 1, (1962) p. 384-386, hereby incorporated by reference). The protocols of the oxidation reaction are reproduced in Tabl...

example 2

Binding of HES with Oxidized Reducing End Groups to HSA in the Aqueous Phase

[0147] For the coupling, hydroxyethyl starch with oxidized reducing end groups (ox-HES) and HSA were completely dissolved in water. When the solution was clear, EDC dissolved in water was added. After activation by EDC accompanied by moderate stirring, further quantities of EDC were added. Where appropriate, the reaction was activated with HOBt and left to stand overnight. The product was purified for 15 hours by dialysis against distilled water and subsequently lyophylized (called Process A in the following).

[0148] The protocols of the coupling reaction are in Table 2.

TABLE 2Coupling reactions between HES (130 kD and 10 kD) with oxidized reducing endgroups and HSA under different conditions (Process A; number in brackets in theHES column reproduces the oxidation process according to Table 1).ox-HESActi-Process AHSA(Mn)EDCHOBtSolventvationReactionCoupling I300mg100mg (1)25mg100mgH2O / dioxane1.5 hours4 hou...

example 3

Direct Binding of HES to HSA in the Aqueous Phase

[0150] The principle of this reaction is based on the formation of Schiff s bases between HES and amino groups of the protein, the reaction being controlled through the reaction of the Schiff s bases to the corresponding amine by NaBH4 (called Process B in the following).

[0151] For this, HES was dissolved completely in a small amount of water. To this end, HSA dissolved in borate buffer, pH 9.0, was added. NaBH4 was added to this solution and the whole left at room temperature accompanied by stirring. A further aliquot of HES 130 kD, followed by further NaBH4, was added. After the reaction was finished, dialysis and freeze-drying was carried out as described.

[0152] The protocols of the individual tests are summarized in Table 3.

TABLE 3Direct coupling between HES (130 kD and 10 kD) and HSA under differentconditions (Process B).Process BHSAHES (Mn)NaBH4Buffer pHReaction timeCOUPLING I50mg500mg500mgNa2HPO4, 0 ml48 hoursHES 1307.5 × ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
average molecular weightaaaaaaaaaa
average molecular weightaaaaaaaaaa
MWaaaaaaaaaa
Login to view more

Abstract

The invention relates to compounds comprising a conjugate of hydroxyalkyl starch (HAS) and an active agent, whereby the hydroxyalkyl starch is either directly covalently bonded to the active agent, or by means of a linker. The invention further relates to methods for the production of a covalent HAS-active agent conjugate, whereby HAS and an active agent are reacted in a reaction medium, characterised in that the reaction medium is water or a mixture of water and an organic solvent, comprising at least 10 wt. % water.

Description

[0001] The present invention relates to compounds comprising a conjugate of hydroxyalkyl starch (HAS) and an active ingredient, wherein the hydroxyalkyl starch is coupled to the active ingredient either directly or via a linker. The invention further relates to processes for the preparation of a covalent HAS-active ingredient conjugate in which HAS and an active ingredient are reacted with each other in a reaction medium, wherein the reaction medium is water or a mixture of water with an organic solvent, having at least 10 weight-% water. The invention further relates to the medical use of the conjugates. TECHNICAL BACKGROUND [0002] The clinical use of many active ingredients of pharmaceuticals is adversely affected by a number of problems (e.g. Delgado et al., Critical Reviews in Therapeutic Drug Carrier Systems, Vol. 9 (3, 4), (1992) pp. 249-304). Parenterally administered native proteins are subject for example to excretion by the reticuloendothelial system, the liver, the kidney...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(United States)
IPC IPC(8): A61K48/00A61K38/28A61K38/22A61K38/21A61K38/19A61K38/18A61K38/16C07J17/00C07H21/02C07K14/56A61K38/00A61K45/00A61K47/36A61K47/48A61P3/02A61P3/04A61P3/10A61P5/00A61P5/44A61P5/50A61P7/00A61P7/06A61P9/06A61P11/06A61P13/00A61P13/12A61P23/00A61P23/02A61P25/24A61P29/00A61P31/00A61P31/04A61P31/06A61P31/10A61P31/12A61P33/00A61P35/00A61P35/02A61P35/04A61P37/04A61P37/08A61P43/00
CPCA61K47/4823A61K47/48284A61K47/61A61K47/643A61P11/06A61P13/00A61P13/12A61P23/00A61P23/02A61P25/24A61P29/00A61P3/02A61P3/10A61P31/00A61P31/04A61P31/06A61P31/10A61P31/12A61P33/00A61P35/00A61P3/04A61P35/02A61P35/04A61P37/04A61P37/08A61P43/00A61P5/00A61P5/44A61P5/50A61P7/00A61P7/06A61P9/06A61K47/50
Inventor SOMMERMEYER, KLAUSEICHNER, WOLFRAMFRIE, SVENJUNGHEINRICH, CORNELIUSSCHARPF, ROLANDLUTTERBECK, KATHARINA
Owner FRESENIUS KABI DEUT GMBH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap