HK1111344A - Stabilized freeze-dried formulation for cephalosporin derivatives - Google Patents

Description

Stable freeze-dried preparation of cephalosporin derivative

The present invention relates to a freeze-dried formulation of a cephalosporin derivative with increased stability, a solution for obtaining said formulation and a process for preparing said formulation, as well as the use of certain compounds for increasing the stability of a cephalosporin derivative in a freeze-dried formulation.

It is known that the lyophilization process may have some effect on the degradation of the pharmaceutically active ingredient in the formulation, as well as a strong effect on the stability of the active ingredient in lyophilized form. The influencing factors mainly comprise pH, salt content, types and contents of excipients in the preparation, selection of temperature, pressure and duration in the freezing process, and operation of sublimation and drying.

Amino acids, polyols are often used to maintain the stability of lyophilized products; however, a large number of relevant documents are available which do not give an answer to how to obtain a stable lyophilized pharmaceutical preparation.

More specifically, according to the teachings of the literature, the presence of an amino acid or a polyhydroxy compound such as mannitol, in the crystalline or amorphous phase, although having certain advantages, has the disadvantage of shortening the useful life of the lyophilized product and/or of requiring low storage temperatures if the lyophilized product contains particularly sensitive active ingredients such as cephalosporin derivatives.

The effects of polyols and amino acids, respectively, have been studied for Human Growth Hormone (hGH), but no good description has been given of their synergistic effects (Pikal M.J., Dellermann K.M., Roy M.L., Riggin M.N., The effects of formulations on The stability of freeze-dried Human Growth Hormone, pharm.research, 1991, 8, No.4, 427-) 436).

The advantages and disadvantages of amino acids and mannitol are as follows:

the advantages of the presence of amino acids:

it has been demonstrated that the presence of glycine in the lyophilized product induces crystallization of molecules in solution during the freezing phase of the lyophilization process (Korey d.j., Schwartz j.b., Effects of excipients on crystallization of pharmaceutical compounds during lyophilization), j. parederal sci.tech., 1989, 43 (2): 80-83). Crystallization of the active ingredient is beneficial to enhance its stability.

In the crystalline state, alanine is advantageous in preventing the collapse of the lyophilized product during sublimation and drying, or in providing a lyophilized product with a larger specific surface area in production, thereby making the drying process more rapid (Pikal M.J., Freeze-drying of proteins, Biopharm., 26-30 October 1990).

The disadvantages of amino acids:

the addition of amino acids to a solution of a sugar or polyol to be lyophilized generally lowers the glass transition temperature of the sugar (te boost m.p.w.m., de Ruiter r.a., de Meere a.l.j., Evaluation of the physical stability of a sucrose-containing lyophilized formulation by differential scanning calorimetry, pharm. research, 1992, 9, 109-. It is now generally accepted that a decrease in the glass transition temperature means a decrease in the stability of the Freeze-dried product (Franks F., Freeze-drying; from empiricism to predictive (Freeze-drying; from empirical to predictive), Cryo-letters, 1990, 11, 93-110).

Advantages of the presence of mannitol:

mannitol present in the lyophilized protein product generally functions as a bulking agent, i.e., it maintains the solid rigid structure volume of the lyophilized product comparable to the volume of the solution to be lyophilized. But its presence may also adjust the osmotic pressure of the reconstituted solution prior to injection. When mannitol is the primary excipient of a Lyophilized protein product, it is usually present in the crystalline state (lyophilised formulations of recombinant tumor necrosis factor, Horam. S., Rana R.K., Smith F.W., pharm. Res., 1992, 9(1), 33-36).

The disadvantages of mannitol:

it has been reported that methylprednisolone sodium succinate in lyophilized form, in the presence of mannitol, is hydrolyzed to a greater extent than in the presence of lactic acid, and that as the amount of mannitol in the lyophilized product increases, the degree of hydrolysis increases. This phenomenon is explained by the fact that during the lyophilization process, the formation of mannitol crystals changes the distribution of water in the matrix of the lyophilized product. The resulting increase in water in The microenvironment of The active ingredient accelerates The hydrolysis of The active ingredient, which leads to a decrease in its stability (The effect of The bulking agent on The solid state stability of lyophilized methylprednisolone sodium succinate), hermann b.d., Sinclair b.d., Milton n.l., pharma.res, 1994, 11(10), 1467-.

The kinetics of degradation of cephalosporin derivatives in aqueous solutions of various carbohydrates, polyhydric alcohols show that degradation is accelerated as the concentration of hydroxyl compounds increases (the effect of carbohydrates and polyhydric alcohols on the stability of aqueous cephalosporin solutions), Hans Bundgaard, Claus Larsen, int. journal of pharmaceuticals, October 1983, 16, 3, 319-.

In addition, in freeze-drying, mannitol is generally used as a carrier or constituting medium to make the freeze-dried product uniform, stable and have a good appearance. The role of mannitol as a stabilizer is unknown for non-protein compounds, in particular cephalosporin derivatives (see, e.g., Handbook of pharmaceutical Excipients, Rowe, R.C., Sheskey, P.J., Weller, P.J, 4 th edition, Php, Londong, 373-377).

In summary, the scientific literature gives contradictory information about the effect of excipients of different properties on the stability of pharmaceutical active ingredients, and a causal relationship between the structure of a lyophilized product and its stability cannot be obtained therefrom. Likewise, the action of the polyhydroxyl compounds and of the amino acids, whether used alone or together, does not allow a series of rules to be concluded, but contradictory results have been observed according to the quantities of active principle and of excipients studied.

Based on the above prior art it was an object of the present invention to provide a freeze-dried formulation of a cephalosporin derivative which shows an increased stability during production and/or later storage.

This problem is solved by the lyophilized formulation of claim 1, the solution for preparing a lyophilized formulation of claim 14, the method of preparing said formulation of claim 15, the use of certain stabilizers of claim 17.

The present inventors have surprisingly found that certain substances have an unexpected stability-enhancing effect on freeze-dried formulations of cephalosporin derivatives.

The invention therefore relates to a pharmaceutical preparation in the form of a lyophilized product, which contains at least one cephalosporin derivative as active ingredient and at least one stabilizer selected from the group consisting of carbohydrates, polyhydric alcohols and polyvinylpyrrolidone (PVP).

In the formulations of the present invention, the cephalosporin derivative may remain stable at 25 ℃ or even higher, thereby increasing its useful life.

The stabilization of the cephalosporin derivative by the formulations of the invention includes stabilization during the manufacturing process.

In a particularly preferred embodiment, the stabilizer is selected from mannitol, trehalose and PVP.

In the present invention, the active ingredient contained in the preparation may be a single active ingredient or may coexist with other proteinaceous or non-proteinaceous antibiotic active ingredients.

In addition to the carbohydrate, polyhydric alcohol and/or PVP, the formulation may also comprise one or more compounds selected from buffers, amino acids, acids or bases to adjust pH, surfactants, salts, preservatives, antioxidants, chelating agents.

Although buffers and amino acids may have additional stabilizing effects, the other ingredients mentioned above are well known pharmaceutically acceptable excipients commonly used in lyophilized formulations. Other conventional additives known to those skilled in the art of pharmaceutical formulation, such as flavoring agents, dyes, etc., may also be added.

As buffers which may be added to the formulations of the present invention, mention may be made in particular of citrate, tris (hydroxymethyl) aminomethane, maleate, succinate, tartrate, carbonate, bicarbonate buffers, and monobasic acid buffers, such as lactate, glycine or acetate buffer systems. It will be appreciated that the acids and bases comprising the buffer may also be added separately, including hydrates, or in any combination thereof.

As surfactants which can be added to the formulations of the invention, mention may be made of polysorbates, poloxamers, tyloxapol, lecithins.

As salts which can be added to the formulations of the present invention, there may be mentioned in particular sodium salts, such as sodium edetate (EDTA tetrasodium salt), sodium chloride, sodium octylbenzenesulfonate (sodium 1, 4-di (2-ethylhexyl) sulfosuccinate), sodium hydrogencarbonate, sodium glutamate, potassium acetate, potassium carbonate and magnesium stearate.

As preservatives which may be added to the formulations of the present invention, there may be mentioned in particular methyl and propyl parabens, benzethonium chloride, thimerosal, phenylmercuric nitrate, benzyl alcohol, phenol and m-cresol.

The formulations of the present invention may be reconstituted into liquid form by addition of suitable solvents or reconstituting solutions for administration by parenteral, intramuscular or oral routes, or may be administered directly to a human or animal by oral routes. Alternatively, liquid or dry formulations may be administered by inhalation.

The cephalosporin derivatives of the present invention include all pharmaceutically acceptable salts and polymorphs as well as hydrates. Furthermore, the term cephalosporin derivative also includes drugs and prodrugs.

In a preferred embodiment, the formulation of the invention comprises a cephalosporin derivative of the following general formula I:

wherein:

R¹is hydrogen, C optionally substituted by fluorine_1-6-alkyl, or C_3-6-a cycloalkyl group;

R²is hydrogen or a group selected from: -CH₂(＝CHR)-COOR、-CH₂OCOR、-CH(R)OCOR、-CH(R)OCOOR、-CH(OCOR)OCOR、-CH₂COCH₂OCOR and

R³is hydrogen or a group selected from: -CH₂C(＝CH₂)-COOR、-COOCH₂C(＝CHR)-COOR、-COOCH₂OCOR、-COOCH(R)OCOR、-COOCH(R)OCOOR、-COOCH(OCOR)OCOR、-COOCH₂COCH₂OCOR and

provided that R is²And R³One is hydrogen and R²And R³Is not hydrogen;

r is hydrogen or C_1-6-an alkyl group;

R⁴is hydrogen or hydroxy;

R⁵is hydrogen or omega-hydroxyalkyl; and is

X is a group selected from the group consisting of CH and N,

as well as pharmaceutically acceptable salts and polymorphs of said compounds and hydrates of the compounds of formula I and salts thereof.

The stability of the compounds of formula I above in the preparation of lyophilized formulations is known to be a problem, as described in EP 1087980B 1.

A particularly preferred example of a compound of formula I is (6R, 7R) -7- [ (Z) -2- (amino- [1, 2, 4)]Thiadiazol-3-yl) -2-hydroxyimino-acetamido]-3- [ (E) - (3 'R, 5' R) -5 '-hydroxymethyl-1' - (5-methyl-2-oxo- [1, 3)]Dioxolen-4-ylmethoxycarbonyl-2-oxo- [1, 3']Bipyridin-3-ylidenemethyl-8-oxo-5-thia-1-aza-bicyclo [4.2.0]Oct-2-ene-2-carboxylic acid, in which R¹、R²、R⁴And R⁵Are each hydrogen, R³Comprises the following steps:

and R is methyl. Hereinafter, this compound is referred to as BAL 5788.

The formulation of the invention may be obtained by freeze-drying an aqueous solution containing at least one cephalosporin derivative as active ingredient and at least one stabilizer selected from the group consisting of carbohydrates, polyhydric alcohols and PVP. In particular, the stabilizer is selected from mannitol, trehalose and PVP.

The concentration of the stabilizer in the solution is preferably in the range of 5 to 80% by weight. Specifically, the concentration of mannitol, trehalose and PVP ranges from 2 to 40% by weight. A particularly preferred concentration range is 10 to 25% by weight. In this application, weight percentages always refer to dry weight.

The solution may also contain pharmaceutically acceptable buffers for further stabilization and/or for adjusting the pH, such as citrate, tartrate, carbonate, bicarbonate, lactate, glycine, acetate or succinate buffers.

The preferred pH range is 2.0 to 6.5, with pH 4.0 to 5.0 being most preferred.

The amount of active ingredient is determined by its solubility in aqueous solutions. The formulations of the invention are obtained by freeze-drying an aqueous solution in which the active ingredient is preferably dissolved.

Particularly preferred solutions for forming the formulations of the present invention comprise a cephalosporin derivative of formula I, the stabilizer mannitol and an aqueous citric acid buffer.

The solution for freeze-drying is prepared, for example, as follows:

the required amounts of active ingredient, stabilizer, buffer and other optional additives such as preservatives are added to the injectable amount of water or solubilizing amount of solubilizer at the appropriate dissolution temperature until fully dissolved. The resulting solution is sterile filtered and filled into containers, preferably vials or capsules.

The freeze-drying of the solution can be carried out as follows:

the solution was subjected to a cycle comprising the following process: frozen and then sublimed and dried according to the volume of freeze-drying and the container holding the solution.

The time, temperature and pressure of sublimation and drying need to be adjusted according to the volume of solution to be lyophilized and the residual amount of water required in the lyophilized product.

The present invention will be described by way of specific examples, which do not limit the scope of the present invention.

The following solutions were prepared, lyophilized, and the resulting formulations were tested for stability.

Preparation of solution a (containing mannitol):

solution ｃA was prepared by dissolving 192.0 g BAL 5788 (synthetic reference EP- ｃA-1087980) and 34.38 g mannitol (from Roquette americｃA, Inc.) in ｃA sodium hydroxide/citric acid buffer system (prepared by dissolving 2.18 g citric acid monohydrate in WFI (water for injection) and then adjusting the pH with sodium hydroxide) at ｃA pH of 4.2-4.8 to ｃA total weight of 1389.5 g. The solution was filtered and filled into vials.

Freeze-drying of solution a:

solution a was freeze-dried under the conditions summarized in the following table to obtain formulation a:

freeze-drying stage	Shelf temperature C	Pressure [ μ bar [ ]]
			Freezing	About-48	Atmospheric pressure
Drying for 1 time	Between-40 and-30	About 70 or less
			Drying for 2 times	-30 to 30	About 70 or less

Preparation of reference solution B (without mannitol):

reference solution B was prepared in the same way as solution a: the same amount of BAL 5788 was dissolved in the same sodium hydroxide/citric acid aqueous injection buffer except that mannitol was not added. The solution was filtered and filled into vials.

The reference solution was lyophilized under the conditions listed above to obtain formulation B.

The lyophilized products made from solution a and solution B were analytically characterized and the stability testing procedure was initiated at different temperatures.

Components of preparation A and preparation B (nominal values)

Composition of formulation a (per bottle):

compound (I)	Weight [ mg ]]
		Bal 5788	999.8
Citric acid monohydrate	15.9
		Mannitol	179.1
Sodium hydroxide is added to	pH 4.2-4.8
		Water for injection	≤3％

Composition of formulation B (per bottle):

compound (I)	Weight [ mg ]]
		Bal 5788	999.8
Citric acid monohydrate	15.9
		Sodium hydroxide is added to	pH 4.2-4.8
Water for injection	≤3％

As a result:

the amount of degradation products formed during the production process was about 9% lower for formulation a (with mannitol) compared to formulation B (without mannitol).

During storage (5 ℃, 12 months), the amount of degradation products formed in the freeze-dried product prepared from solution a (containing mannitol) was about 10% lower than the amount formed during storage of the freeze-dried product prepared from solution B (without mannitol).

Other formulations were prepared in the same manner with trehalose or PVP and with other buffer systems.

Claims (18)

1. A freeze-dried formulation of a pharmaceutically acceptable cephalosporin derivative, which comprises at least one cephalosporin derivative as an active ingredient and at least one stabilizer selected from the group consisting of carbohydrates, polyhydric alcohols and polyvinylpyrrolidone (PVP).

2. The lyophilized formulation of claim 1, wherein the carbohydrate is trehalose.

3. The lyophilized formulation of claim 1, wherein the polyhydric alcohol is mannitol.

4. The lyophilized formulation of any one of claims 1 to 3, further comprising an additional antibacterial active ingredient.

5. The lyophilized formulation of any one of claims 1 to 4, further comprising one or more compounds selected from the group consisting of: buffers, amino acids, acids or bases to adjust pH, surfactants, salts, preservatives, antioxidants, chelating agents.

6. A lyophilized formulation of any one of claims 1 to 5 for reconstitution into a solution for administration by parenteral route, intramuscular route, oral route or by inhalation; or for direct administration by the oral route or by the inhalation route.

7. The lyophilized formulation of any one of claims 1 to 6, wherein the cephalosporin is a compound of formula I:

wherein:

R¹is hydrogen, C optionally substituted by fluorine_1-6-alkyl, or C_3-6-a cycloalkyl group;

R²is hydrogen or a group selected from: -CH₂(＝CHR)-COOR、-CH₂OCOR、-CH(R)OCOR、-CH(R)OCOOR、-CH(OCOR)OCOR、-CH₂COCH₂OCOR and

R³is hydrogen or a group selected from: -CH₂C(＝CH₂)-COOR、-COOCH₂C(＝CHR)-COOR、-COOCH₂OCOR、-COOCH(R)OCOR、-COOCH(R)OCOOR、-COOCH(OCOR)OCOR、-COOCH₂COCH₂OCOR and

provided that R is²And R³One is hydrogen and R²And R³Is not hydrogen;

r is hydrogen or C_1-6-an alkyl group;

R⁴is hydrogen or hydroxy;

R⁵is hydrogen or omega-hydroxyalkyl; and is

X is a group selected from the group consisting of CH and N,

as well as pharmaceutically acceptable salts and polymorphs of said compounds and hydrates of the compounds of formula I and salts thereof.

8. The formulation of any one of claims 1 to 7, obtained by freeze-drying a solution containing at least one cephalosporin as active ingredient, at least one stabilizer selected from the group consisting of carbohydrates, polyols and PVP, and an aqueous solution.

9. The formulation of claim 8 wherein the stabilizer is mannitol, trehalose or PVP.

10. The formulation of claim 8 or 9, wherein the aqueous solution is a buffer solution, preferably a citrate, tartrate, carbonate, bicarbonate, succinate, glycine, lactate or acetate buffer.

11. The formulation of any one of claims 8 to 10, wherein the aqueous solution is a monobasic acid buffer, such as an acetate, glycine or lactate buffer.

12. The formulation of any one of claims 8 to 11, wherein the concentration of the stabilizing agent in the solution is 5-80% by weight.

13. The formulation according to any one of claims 8 to 12, wherein the pH of the solution is 2.0-6.5, in particular 4.0-5.0.

14. Solution for the preparation of a lyophilized formulation comprising, in aqueous solution, at least one cephalosporin derivative as active ingredient, at least one stabilizer selected from the group consisting of mannitol, trehalose and PVP.

15. A process for the preparation of a stable, pharmaceutically acceptable lyophilized formulation of a cephalosporin derivative comprising two steps:

(a) adding at least one stabilizer selected from the group consisting of carbohydrates, polyhydric alcohols and PVP to an aqueous solution of a cephalosporin derivative; and is

(b) The above solution was lyophilized.

16. The method of claim 15, wherein mannitol, trehalose or PVP is added as a stabilizer.

17. Use of a compound selected from the group consisting of a carbohydrate, a polyhydroxyl alcohol and PVP for stabilizing a cephalosporin derivative in a lyophilized formulation.

18. Use according to claim 17, characterized in that the compounds are mannitol, trehalose and PVP.