CA2151409C - Formulation for stabilizing human immunoglobulins - Google Patents

Abstract

The invention relates to a formulation for stabilizing human immunoglobulins such as human anti-Rho [D] immunoglobulins. The solution is provided in an injectable solution at 25°C and comprises human serum albumin, sodium phosphate dihydrogen in a sufficient quantity to maintain a pH between 5 and 7.2 and a saccharide selected from the group consisting of glucose, saccharose, maltose, lactose and mixtures thereof. It can be applied either intravenously or intramuscularly. The invention is likewise related to the use of the formulation.

Description

Background of the Invention Rho heteroimmunization is an autoimmune response which takes place in non sensibilized Rho (D) negative persons, when they receive or have been exposed to blood from olio (D) positive persons. This happens for example in s the parturition of Rho (D) positive children; in this case, if the necessary precautions are not taken an immunological response takes place in Rho (D) negative mothers, which leads to the formation of anti-Rho (D) antibodies so that in the next pregnancy, haemolytic disorders take place in the case of Rho (D) positive pregnancy or foetus, leading to foetus-maternal haemorrhages, during normal parh~rition, spontaneous or indirect abortion, with symptoms and signs of amniocentesis, abdominal trauma, etc.
Rho heteroimmunization also takes place in other situations for example, in Rho (I7) negative persons subjected to transfusions with Rho (D) positive blood or blood components, such as red corpuscules of Rho (D) ~s positive blood.
Conventional treatment in these situations is the prevention of Rho heteroimmunization of mothers or would-be mothers as well as persons at risk of heteroimmuniza.tion or in conditions of immunology with a reduction in blood platelets. Said preventive treatment is based on the administration of 2o gamma immunoglobulin (IgG), according to a predetermined preventive program according; to each case. In the case of non sensibilized Rho (D) negative women, tree preventive treatment requires the administration of human anti-Rho IgG as from the 28th week of gestation, when the progenitor is Rho (D) positive or unknown. With this type of treatment, the risk of Rho 2s heteroimmunization is reduced to 0. 1 %. If said preventive treatment is carried out within ?2 hours of birth, the risk of Rho heteroimmunization is 1 to 2% (compared to 12-13% in ordinary situations), ascribed to Rho heteroimmunization which occurs naturally in the last three months of pregnancy.

The responses of Rho heteroimmunization present delicate clinical symptoms and signs, mainly due to haemophylic (haemorrhages) and haemolytic disorders.
In the above mentioned preventive treatment of Rho s heteroimmunization, a fraction of gamma immunoglobulin from human plasma formulated in injectables is administered intravenously or intramuscularly.
There are various commercial products used in prenatal and obstetric clinic; for example Partogamma T* and the Win Rho~R~ line, which includes the latest product known as Win Rho SD~R~, manufactured by Rh Pharmaceutical Inc., mentioned here as a point of reference. The suppressive agent of Rho heteroimmunization in these products, is a fraction of human IgG, obtained by chromatographic fractionation in columns of anionic exchange of IgG of human plasma, and. then treating the obtained fraction with solvent/detergent so as to inactivate the lipidic membrane of certain virus, such as the Hepatitis C
is virus and HIV.
These products and other similar ones are formulated as lyophylized injectable compositions, which contain glycine and sodium chloride as stabilizing system in sufficient quantity to form a 0.10 and 0.15 M solution respectively, in doses of, for example, 250 to 330 mcg/ml of injectable solution.
Given the general tendency of immunoglobulins to form in solution, * Trade-mark additions or undesirable molecular complexes (dimers, trimers, tetramers) conditional upon more antigenic properties, and anti-complementary activity particularly in the case of intravenous preparations, other stabilizing systems have been proposed such as:
polyethylene glycol human-albumin-manitol; glycine-histidine, inorganic salts-proteins-saccharides, human albumin-glycine-manitol, etc. of variable stability according to the stabilizer. In the case of Rho [D] Immunoglobulin stabilized with glycine, the US Pharmacopia (abbreviation in Spanish: USP) XXIII acknowledges a stability of not more than 6 months, for products of this type stored at temperatures between 2° and 8°C. (page 708).
It has been found, and this constitutes the aim of this invention, that it is possible to prepare and have available new compositions for the prevention of IRho heteroimmunization, formulated with human gamma immunoglobulin, in aqueous solution of reduced tendency to molecular addition, verified for periods of at least 24 months at room temperature.
Concise Description of the invention One the of aims of this invention is a formulation for stabilizing human immunoglobulins. The formulation comprises human serum albumin, sodium phosphate dihydrogen in a sufficient quantity to maintain a pH between 5 and 7.2 and a saccharide selected from the group consisting of glucose, saccharose, maltose, lactose and mixtures thereof .
The immunoglobulins suitable according to the present invention may be for example human anti-Rho [D]
immunoglobulins. These human anti-Rho [D] immunoglobulins may be dissolved in a solution which is injectable at 25°C.
Another aim of this invention is to provide a formulation for stabilizing human immunoglobulins characterized in that the formulation is an aqueous solution comprising per 100 ml of solution:
- 0.5 to 2 g of human serum albumin;
- 4 to 6 g of the saccharide; and - sodium phosphate dihydrogen in a sufficient quantity to maintain a pH between 5 and 7.2.
Preferably, the formulation may contain 1.0 g of human serum albumin, 4 to 6 g of the saccharide and 0.28 g of sodium phosphate dihydrogen. The formulation is preferably an aqueous solution which is formulated in an aqueous vehicle. This aqueous vehicle may in turn be formulated in intravenous or intramuscular injectable units.
Another aim of this invention is to provide a use of the formulation in accordance with the present invention for the stabilization of human immunoglobulins.
A

. 2151409 Detailed description of the invention The compositions of the present invention are the result of lengthy investigations and trials carried out with the aim of finding preventive pharmaceutical compositions for Rho Heteroimmunization and other clinical symptoms and signs such as Immuno Purpura thrombocytopaenia, cited before,capable of withholding during prolonged periodsthe suppressing capacity of immuno-response in affected persons.
In practice, and according to the preceding text, the usual treatment is based on the administration of compositions formulated with Rho [D] human IgG in vials of intravenous or intramuscular application. As IgG has a tendency to form antigenic molecular additions, conventional stabilized formulations come in lyophilized vials, being restorative in situ. Therefore, the cost of this type of products rises, conditioned upon overloading due to breaking up and transport. This has led to the development of stabilized formulations of anti-Rho [D] IgG
of this invention, capable of withholding its immunocapacity during prolonged periods and without signs of side effects.
A,.

_. ~ -6- r 251409 Therefore, the compositions of this invention fit in within this line and these aims, formulated by resorting to an unpublished combination of stabilizing components capable of ensuring a prolonged permanence of extended anti-Ro (D) activity, according to tests carried out during at least s 24 months.
In the previous texts as well as in the text which follows it will be understood that the term human Rho (D) immunoglobulin refers to globulin originating from human plasma which contains antibodies for the Rho (D) factor, as stated by the US Pharmacopia XXIII Ed (page 708) io and equivalent products.
It should likewise be understood that the term serum albumin refers to the albumin projections taken from the blood of healthy donors as defined in the US Pharmacopeia XXIII Ed. page 38 and equivalent products.
The stabilizing system of the compositions of this invention includes:
- saccharides such as glucose, saccharose, lactose, or maltose, mixtures of same, in concentrations of 4 to 6% the weight to volume, preferably 4.5 to 5.8% weight to volume, more preferably 4.9-5.1 weight to volume;
- serum albumin in concentrations of 0.5 to 2% weight to volume, preferably 0.8 to 1.5% weight to volume, particularly 0.9 to 1.1.%
weight to volume, free of alpha-globulin and bilirubin;
- sodium phosphate dihydrogen in sufficient quantity to form a pH
:~ :~
.~._t ~' -' - ~ 2 1 5 1 4 0 9 between 5 and 7.2, preferably between 5.0 and 6, depending on the concentration of IgG and albumin, added as NaH2P04 or formed from, for example, mixtures of Na2HP04 or Na3P04 and orthophosphoric acid.
For. example, for a formulation in accordance with the invention, which s contains 1.0 g of human serum albumin, 5.0 g of saccharose and 2500 micrograms of anti-Ro (D) IgG taken up to a volume of 100 ml with water reaches an optimum pH of 5.1-5.4 with 0.28 g of monohydrate sodium phosphate dihydrogen.
The pharmaceutical compositions of this invention can easily be to stored at room temperature in dosed units (vials) for immediate use, or in bigger containers for transport, later use, or to be lyophylized if necessary.
In all cases in the previous paragraph, the term human plasma and products extracted from human plasma such as Ro (D) positive IgG, refer to plasma and/or serum albumin and IgG tested in order to verify the is presence of HIV, HCV ~ and HbsAg antibodies (surface antigen for Hepatitis B virus).
The following example describes and summarizes the tests and results obtained in the study and verification of the stability and biological activity of the composition of this invention.
2 o Batches studied 2 batches of products in two different concentrations were prepared:
(abbrev.in sp.:PS) 250 mcg (abbrev.in sp.:LP1) PS 330 mcg LP1 _g_ Tests carried out Variations with time of the following parameters were assessed:
i) Appearance of solution ii) pH of solution a iii) Molecular distribution (abbrev. in sp.:H.P.L.C.) iv) Anti-Rho activity Conclusions The results obtained show the good stability of the stabilized solution in to accordance with this invention.
No significant changes in product activity were observed, nor the appearance of dimerous, additions :nor proteins of low molecular weight.
Results obtained l a The following Tables 1 and 2 summarize the results of tests carried out.
Table 1 illustrates results of tests carried out on the following composition:
Composition of this invention PS 250 mcg LP I
Table 1 Natural stability trial Time Molecular Distribution (months)Appearance pH Dimers MonomersOthers Activity 0 Colourless 5.2 0.2% 99.8% - 300 mcg solution 6 without change5.2 0.2% 99.8% - 306 mcg 12 without change5.1 0.5% 99.5% - 290 mcg 24 without change5.1 1.2% 98.8% - 303 mcg Table 2 illustrates results of tests carned out on the following composition:
Composition of this invention PS 330 mcg LP1 Table 2 Natural stability trial Time Molecular ion Distribut (months)A earance H Dimers Monomers Others Activi 0 Colourless 5.2 0.4% 99.6% - 363 mcg solution 6 without change5.2 0.6% 99.4% - 333 mcg 12 without change5.1 1.0% 99.0% - 363 mcg 24 without change5.1 1.5% 98.5% - 333 mcg The term "effective quantity" as used here, represents a quantity of y Rh (D) positive human IgG, capable of blocking Rho heteroimmunization in Rho (D) negative persons. The particular doses administered in accordance with this invention, are determined by particular circumstances of each case.
The term "treatment", as used here describes the mode and care of patients with the aim to prevent the onset, ease symptoms and/or follow-up the evolution and development of Rh heteroimminization and/or other situations which might requiere the application of Rh (D) positive human IgG.
The compositions of this invention are prepared in accordance with known techniques using human Rh (D) IgG, human serum albumin and remaining components, all known and easily available.

Claims (6)

1. Formulation for stabilizing human immunoglobulins characterized in that said formulation comprises human serum albumin, sodium phosphate dihydrogen in a sufficient quantity to maintain a pH between 5 and 7.2 and a saccharide selected from the group consisting of glucose, saccharose, maltose, lactose and a mixture thereof.

2. The formulation of claim 1, wherein said human immunoglobulins are human anti-Rho[D]immunoglobulins.

3. The formulation of claim 2, wherein the formulation is dissolved in a solution which is injectable at 25°C.

4. The formulation of claim 1, 2 or 3, wherein said formulation is an aqueous solution comprising per 100 ml of the solution:
- 0.5 - 2 g of human serum albumin;
- 4 - 6 g of the saccharide: and - sodium phosphate dihydrogen in a sufficient quantity to maintain a pH between 5 and 7.2.

5. The formulation of claim 1, 2, 3 or 4, wherein said aqueous solution is formulated in an aqueous vehicle, said aqueous solution comprising per 100 ml of the solution:
- 1.0 g of human serum albumin;
- 4 - 6 g of the saccharide; and - 0.28 g of sodium phosphate dihydrogen, wherein said aqueous vehicle is formulated in intravenous or intramuscular injectable units.

6. Use of a formulation according to claim 1, 2, 3, 4 or 5 for the stabilization of human immunoglobulins.