KR20090106666A - Compositions containing organic compounds - Google Patents

Abstract

Pharmaceutical formulations comprising en epothilone in the form of an infusion concentrate or a lyophilised composition, and methods of administration of an epothilone in suitable form for parenteral administration.

Description

Compositions containing organic compounds

The present invention relates to pharmaceutical preparations of epothilones, in particular parenterally, for example intravenously administrable pharmaceutical preparations.

Epothilones represent microtubule-stabilizing cytotoxic agents of Formula I (Gerth, K et al., Antibiot. 49, 560-3 (1966); or Hoefle et al., DE 41 38 042). Typical representatives include Epothilone A where R is hydrogen and Epothilone B where R is a methyl group.

They are seven asymmetric center containing 16-membered macrolides and are also characterized by various functional groups. For example, they may include other ring systems, such as epoxide and / or thiazole rings. They may have two free, derivable hydroxyl groups, and the macrolide itself may contain ester bonds. Epothilones and their synthesis are described, for example, in published PCT Application Nos. WO 93/10121 and DE 41 38 042 A2, the contents of which are incorporated herein by reference. Typical epothilone derivatives and their synthesis are described in published PCT applications Nos. WO 97/19086 and WO 98/25929, the contents of which are incorporated herein by reference. Reference to epothilones preferably means epothilone A or epothilone B or salts thereof and suitable derivatives or mixtures thereof. Epothilone A or B can be used alone or in a mixture of A and B, but preferably A alone or B alone, most preferably B alone.

Cytotoxic agents are well known for the treatment of tumors. Many of the anti-tumor activities in these compounds rely on cell proliferation inhibition and the resulting induction of apoptosis and apoptosis. Many cytotoxic agents exert their effects through inhibition of DNA and / or RNA synthesis. However, for certain cytotoxic agents such as members of the taxane family, such as paclitaxel, and epothilones, their activity depends on inhibition of microtubule growth. Microtubules are an important and attractive target for the development of new anticancer agents.

However, little is known about agents suitable for epothilones. We found that the 16-ring macrolide system was particularly susceptible to degradation. Moreover, the low solubility of these compounds makes the formation of pharmaceutical preparations for parenteral administration very difficult. Poorly soluble compounds can typically be solutioned by heating the solvent during the dissolution process. However, under the high reactivity of these compounds, they can readily degrade at elevated temperatures. Furthermore, these highly reactive compounds can be degraded during long-term storage as aqueous solutions. A concentrated solution of microcontrolled Taxol ^R is described which can be diluted in an aqueous medium prior to intravenous administration. However, such solutions typically use surfactants such as Cremophor ^R (polyethoxylated castor oil). It is well known that surfactants such as Cremophor ^R can induce an allergic reaction in a patient.

Accordingly, there has been a need for commercially acceptable pharmaceutical formulations suitable for epothilones, for example, pharmaceutical formulations that are storeable (eg, in refrigerators (eg, at 2-8 ° C.)).

We now enhance the solubility of epothilones such as epothilone A or epothilone B and / or use surfactants such as surfactants having an HLB level of 10 or more, such as Cremophor ^R. They found ways to dissolve them more quickly without negatively affecting their efficacy.

Accordingly, the present invention provides a pharmaceutical formulation comprising, in one aspect, an epothilone, for example, epothilone A or epothilone B, which may be referred to as a preparation of the present invention.

Provided are methods for enhancing the solubility of epothilones and / or dissolving them more rapidly without negatively affecting their efficacy without the use of surfactants such as surfactants with HLB levels above 10.

In a preferred embodiment the invention provides a pharmaceutical preparation in the form of an infusion concentrate comprising epothilone and a pharmaceutically acceptable organic solvent, for example in the absence of a surfactant having an HLB level of 10 or greater, for example, Cremophore ^R. Provide pharmaceutical preparations. Injection concentrates do not require the use of surfactants to enhance the solubility of epothilones such as epothilones A and B and / or to dissolve them more quickly. As noted above, surfactants such as, for example, polyhydrogenated natural or hydrogenated castor oils, for example HLB levels above 10, for example Cremophore ^R , can cause allergic reactions and they can also be used in standard PVC containers, A plasticizer can flow out from piping etc. As a result, when using them, it will be necessary to use special injection devices such as nitroglycerin tubing and non-plastic containers such as glass, tubing and the like.

The pharmaceutically acceptable organic solvents described above may be selected from all such organic solvents known in the art. The solvents can be used individually or in combination with other solvents. Preferably the solvent is a liquid at room temperature. Preferably the solvent is (i) an alcohol of at least 2 carbon chain lengths, for example C ₂ -C ₅ , for example C ₂ or C ₃ or C ₄ , or (ii) N-alkylpyrrolidone , For example C ₁ -C ₄ , for example N-methylpyrrolidone. Typical examples of alcohols are, for example, water miscible alcohols such as anhydrous alcohols, or glycerol. Other alcohols include glycols, for example all glycols obtainable from oxides such as ethylene oxide, for example propylene glycol. Other examples are polyols such as polyalkylene glycols such as poly (C _2-3 ) alkylene glycols. Typical examples are polyethylene glycols, for example, preferably of molecular weight 200-600 Daltons, more preferably 200-400 Daltons, in particular 300 Daltons. Polyethyleneglycol may be used in distilled form and may, for example, be characterized by one or more of the following features: (iii) an ethylene oxide content of up to 20 ppm, typically less than 1 ppm, for example 0.1 to 0.5 ppm (Ii) pH values of 4.0 to 7.0, and (iii) absence of reducing agents and aldehydes (European Pharmacopoeia, Third Edition, 1997, Council of Europe, Strasbourg, Chapter 2.2.2 Degree of coloration of liquids, incorporated herein by reference). , determined by measuring the degree of pigmentation of the test liquid compared to the control solution comprising iron chloride (yellow solution) or cobalt chloride (red solution) according to the experimental procedures described in p15-17). Those skilled in the art will appreciate that polyethylene glycols of various molecular weights can be used, as long as they are physiologically acceptable. The solvent, of course, contains a residual amount of water arising from their preparation or obtained from the atmosphere, for example, below saturation, eg 2% or less, eg 0.5% or less, eg typically less than 0.1%, eg For example, it may contain 0.01 to 0.05%. If desired, the pharmaceutically acceptable solvent is water ("added water") and, for example, up to 45% water, for example up to 30%, for example 20%, for example 5%. Can be mixed. Typical examples include ethanol / water mixtures such as 70% ethanol weight / volume (w / v), or polyethylene glycol / water mixtures such as 90% polyethylene glycol w / v.

Epothilones, such as epothilones A or B, are from 0.1 to 100 mg / ml, for example from 1 to 100 mg / ml, more preferably from 0.5 to 50 mg / ml, more preferably from 0.5 to 10 mg / ml. May be present in the injection concentrate at a concentration of ml, most preferably 1 mg / ml.

Epothilones such as epothilone A or epothilone B can be used individually or as a mixture of epothilones, for example a mixture of epothilones A and B. Under the more potent anti-tumor activity of epothilone B it can be used at a lower concentration than epothilone A in the formulation. Epothilone A at a concentration of 0.1 to 100 mg / ml, for example 10 to 100 mg / ml, preferably 0.1 to 50 mg / ml, for example 20 to 50 mg / ml, in particular 1 mg / ml, It is preferable to use. Epothilone B, when used alone, is preferably used at a concentration of 0.1 to 50 mg / ml, for example 10 to 50 mg / ml, for example 1 to 50 mg / ml, in particular 1 mg / ml.

Pharmaceutical compositions of the invention in the form of infusion concentrates may be prepared by post-treatment, eg dissolution, of the epothilone in a pharmaceutically acceptable solvent of the invention, optionally with other excipients. Preferably, no other excipients are present. If they are present they are preferably present in an amount of less than 5%, for example less than 2%, for example 0.1 to 1.5%.

The infusion concentrate of the present invention is conveniently stored in a suitable container such as a vial, a double-chamber vial system, or an ampule. Typically vials or ampoules are made of glass, for example borosilicate or soda lime glass. The vials or ampoules may be of any volume customary in the art and are preferably large enough to accommodate 1 to 5 ml, more preferably 2 ml of infusion concentrate. The container may preferably have a punctureable stopper, such as a sterile rubber stopper, which provides a suitable sealed seal to the container to enable the transfer of liquid from or to the container.

Infusion concentrates of the invention may be stable for long periods of time, for example 12 to 36, for example 24 months, at temperatures of at least 2 to 8 ° C., as shown in standard stability tests, for example described in the Examples. .

Moreover, the injection concentrate can be prepared using conventional equipment, which hardly evaporates, eg does not require an anti-explosion device, and withstands rubber stoppers, for example, without disassembling the stopper when stored in the container. I can make it.

Infusion concentrates may be diluted in an pharmaceutically acceptable organic solvent suitable for intravenous administration, such as an aqueous medium, prior to parenteral, eg, intravenous administration, to the patient to form an infusion solution. Parenteral administration is understood to include administration by infusion or injection.

Accordingly, the present invention provides an infusion solution comprising a mixture of an infusion concentrate as defined above and a pharmaceutically acceptable organic solvent, preferably in an aqueous medium.

 Pharmaceutically acceptable organic solvents used as diluents can be either solvents or solvent combinations used in the infusion concentrate. But preferably it consists of water, i.e. water for injection. The infusion solution preferably has the same or essentially the same osmotic pressure as the body fluid. Thus, the diluent preferably contains an isotonic agent (s) that has the effect of making the osmotic pressure of the infusion solution equal or essentially the same as the body fluid.

The tonicity agent (s) can be selected from any known in the art, for example mannitol, dextrose, glucose and sodium chloride. Preferably the tonicity agent is glucose or sodium chloride. The tonicity agent (s) can be used in an amount such that the infusion solution has the same or essentially the same osmotic pressure as the body fluid. The exact amount required may be determined by routine experimentation and may depend on the composition of the injection solution and the nature of the tonicity agent (s). Particular isotonic agent (s) can be selected depending on the nature of the epothilones such as epothilone A or epothilone B. For example, when using epothilone B alone or in combination with epothilone A, the use of certain isotonic agent (s) can lead to turbidity of the infusion solution. Turbidity can result from the dissolution of epothilones such as epothilone B.

Surprisingly, we found that turbidity did not appear at all for long periods of time, for example over 24 hours when using glucose as an isotonic agent.

The concentration of the tonicity agent (s) in the aqueous medium depends on the nature of the particular tonicity agent (s) used, preferably the concentration is 5% or less. When using glucose, it is used at a concentration of 1 to 5% w / v, more particularly 5% w / v. When the tonicity agent is sodium chloride, it is preferably used in an amount of 1% w / v or less, especially 0.9% w / v.

Infusion solutions according to the invention may comprise other excipients commonly used in formulations administered intravenously. Excipients include antioxidants. Antioxidants can be used to protect epothilones such as epothilone B from oxidative degradation. Antioxidants can be selected from any of the antioxidants known in the art and suitable for intravenous preparations. The amount of antioxidant can be determined by routine experimentation. As an alternative or in addition to the addition of antioxidants, the antioxidant effect can be achieved by removing oxygen (air) from contact with the injection solution. This can be conveniently done by purifying the vessel containing the injection solution with an inert gas, for example nitrogen.

The amount of diluent used in admixture with the infusion concentrate to form the infusion solution may be selected according to the desired concentration of epothilone, for example epothilone B, in the infusion solution. Preferably the infusion solution is mixed with a diluent, for example a 5% w / v glucose solution in water for injection, in a suitable container such as an infusion bag or bottle with a vial or ampoule of the above-mentioned infusion concentrate, and the volume is diluted with a diluent. From 50 ml to 1000 ml, for example from 200 ml to §§ 1000 ml, or preferably from 50 to 100 ml. The so formed injection solution can preferably be used immediately or within a short time after formation, for example within 6 hours. Alternatively, the infusion concentrate and the predetermined amount of diluent may each be loaded into separate chambers of the dual-chamber vial system and mixed just prior to intravenous administration to the patient.

In another embodiment the pharmaceutical formulation of the invention may be in the form of a lyophilized composition comprising epothilones such as epothilone A or epothilone B. Under the low solubility of epothilones A and B, the lyophilized mass consisting of only epothilones, epothilones A or epothilones B provides a suitable volume of lyophilized composition, which is convenient for processing, for example, to allow acceptable content uniformity. It can be so small that it can't or even it's hard to see with the naked eye. Thus, excipients which act to increase the volume of the lyophilized composition by increasing the solids content in the lyophilized composition according to the invention can be used. Suitable excipients may be any of the excipients used alone or in combination to increase the volume of the lyophilized composition without negatively interacting with the epothilone or otherwise diminishing its efficacy. In addition, the excipients should be suitable for pharmaceutical preparations, for example parenteral, for example intravenous pharmaceutical preparations. Therefore, the choice of excipient (s) should consider not only the properties of the lyophilized composition, but also the properties of the final pharmaceutical form. Examples of suitable excipients include sodium phosphate or other carbohydrates such as potassium, citric acid, tartaric acid, gelatin, lactose and dextrose, mannitol and dextran and intravenously suitable cyclotextins such as beta-cyclotextrin. Include. Typical beta-cyclodextrins also include beta-cyclotextrin derivatives, such as alkyl- or allyl-derivatives, or hydroxyalkyl-derivatives, for example beta-cyclodextrins and oxides, such as propylene oxide. It is formed by a condensation reaction. In a more preferred embodiment the beta-cyclodextrin derivative can be hydroxypropyl-beta-cyclodextrin. Preferably hydroxypropyl-beta-cyclodextrin is described in Journal of Pharmaceutical Sciences, Vol. 85, No. 11. May be any of those mentioned by Roger A. Rajewski et al. In November 1996, pages 1142-1169.

Through careful selection of excipients, Applicants increase the solubility specific to epothilones such as epothilone A or epothilone B, or dissolve the epothilones more quickly but do not adversely affect the efficacy of the epothilones. It has been found that an appropriate volume of lyophilized composition can be formed, including, for example, Epothilone A or Epothilone B.

The excipients or mixtures thereof may comprise 50 to 99.9%, more preferably 90 to 99%, for example 95% of the total solids of the lyophilized composition.

Epothilones may comprise 100% of the total solids content of the lyophilized composition, but may preferably comprise 0.1-10%, more preferably 0.1-1.5%, for example 1.2% of the total solids.

Among the excipients commonly used in the field of lyophilisates that are reconstituted for pharmaceutical use, such as other excipients mentioned above, to the extent that epothilones and cyclodextrins or mannitol do not provide 100% of the total solids content of the lyophilized composition. Either one can provide a balance of solids.

The lyophilized composition according to the present invention may be formed from a solution containing epothilones such as epothilone A or epothilone B (hereinafter referred to as "stock solution") and suitable excipients as defined above. Suitable solvents for such stock solutions are water based or aqueous base solvents containing pharmaceutically acceptable, water miscible organic solvents such as alcohols, more particularly ethanol or polyethylene glycol.

The stock solution may contain 0.01% to 0.5% (w / v) of epothilones such as epothilone A or epothilone B.

Stock solutions can be prepared by dissolving epothilones such as epothilone A or epothilone B, and excipients in a suitable solvent and then filtering the solution through a filter, eg a sterile 0.22 micron filter. The stock solution so formed can be filled in vials of suitable volume, preferably 30 ml volume and 4.2 ml fill volume.

In another aspect, the present invention provides a process for the preparation of (i) epothilones such as epothilone A or epothilone B in a pharmaceutically acceptable excipient such as mannitol or cyclodextrin, eg hydroxy-beta- Mixing with cyclodextrin to form a stock solution, and (ii) dehydrating the stock solution provides a method for producing a lyophilized composition.

Lyophilization can be carried out according to known techniques. In a preferred method, the filled vial can be frozen in a lyophilization chamber at a temperature below the eutectic point, preferably at about −40 ° C. for about 3 hours. The lyophilization chamber can then be evacuated to about 0.1 to 0.2 millitorr. The temperature of the chamber is then increased to sublimate the freezing liquid. Preferably the temperature is increased to about 0 ° C. and maintained at that temperature for 8 to 15 hours to lyophilize.

The lyophilized composition can be used for the preparation of parenteral preparations so that the lyophilization process is preferably carried out under sterile conditions, for example using aseptic preparation or by irradiation. Sterile solution formulations containing pharmaceutically active compounds, sterile filling of vials and lyophilization procedures under sterile conditions are well known to those skilled in the art.

The moisture content of the lyophilized composition thus formed may be 3% or less of the total weight of the lyophilized composition. However, a humidification step may optionally be used following lyophilization which introduces sterile water vapor into the lyophilization chamber under atmospheric pressure or reduced pressure. Of course, when the humidification step is carried out under reduced pressure, the pressure changes with the introduction of steam, the pressure change can be monitored and, if necessary, the pressure can be adjusted using techniques well known in the art. The humidification step can be completed within 4 to 8 hours depending on whether the humidification step is carried out at atmospheric pressure or at reduced pressure.

The lyophilized composition obtained using the humidification step below is called hydrolyzate. The hydrolyzate may contain 0.1 to 5% by weight of water.

The lyophilized composition according to the invention may be provided in the form of a single dose container. The single dose container form can be any suitable size. "Proper size" means an appropriate size depending on the volume of solution required to reconstitute the lyophilized composition. Any suitable container can be used to provide these formulations. "Proper" means any container that can be used for aseptic filling, maintains a sterile environment and is not reactive with the lyophilized composition. Preferred containers may be formed of glass, for example Type I glass, and may have means for receiving a stopper, for example a sterile rubber stopper, which may provide a sealed seal with the wall of the container. Preferred stoppers may also allow access to the contents of the container for the purpose of introducing a solvent, such as water for injection, to the lyophilized composition.

The lyophilized composition according to the present invention may have a storage stability for 24 to 36 months at a temperature of 2 to 30 ℃. Lyophilized compositions stored during this period show no signs of degradation and their unique solubility is not affected.

If parenteral, eg intravenous, forms of epothilone are desired to be provided, the lyophilized composition may preferably be re-constituted just prior to administration.

Re-constitution may comprise the lyophilized composition with water or some other pharmaceutically acceptable solvent described above, such as physiological saline, pharmaceutically acceptable alcohols such as ethanol, propylene glycol, polyethylene glycol, for example polyethylene glycol. Dissolution in an aqueous solution such as 300, or some other sterile injectable under sterile conditions. The single dose container form may be filled with an appropriate amount of solvent depending on the desired concentration of epothilones such as epothilone A or epothilone B for parenteral administration. Such reconstituted lyophilized compositions may preferably be used immediately or within a short time after formation, for example within 6 hours.

Pharmaceutical formulations of the present invention in a form suitable for parenteral, eg, intravenous administration, for example infusion solutions or reconstituted lyophilized compositions prepared by dilution of infusion concentrates (hereinafter referred to as the dilution preparations of the invention) The container can be filled from any conventional container that is not reactive with the pharmaceutical formulation. It is preferred to use plastic containers, for example plastic infusion bags, but glass containers made of the above-described glass types are suitable.

The plastic container may consist mainly of thermoplastic polymers. The plastic material may additionally include additives such as plasticizers, fillers, antioxidants, antistatic agents and other additives conventional in the art.

Plastics suitable for the dilute formulations of the present invention must be able to withstand the elevated temperatures required for thermal sterilization. Preferred plastic infusion bags are made of PVC plastic materials known in the art.

A wide range of container sizes can be used. In selecting the vessel size, the solubility and ease of operation of the epothilones in certain solvents and, if appropriate, the shelf life of the vessel can be taken into account. Preference is given to using a container which can hold about 200 to 1000 ml, for example 250 to 1000 ml, of the dilution formulation of the invention.

Dilution formulations of the invention are preferably sterile. This can be readily accomplished, for example, by irradiation or by filtering the dilution formulation with a sterile filtration membrane. Aseptic, aseptic filling vials of any composition in liquid form and / or combining parenteral liquids with suitable diluents under sterile conditions are well known to those skilled in the art.

Diluent preparations of the invention are useful for the treatment and prevention of malignant proliferative diseases, for example the signs and conditions disclosed in WO 93/10121 and DE 41 38 042 A2, the contents of which are incorporated herein by reference. More specifically, they are tumor diseases such as melanoma, ovarian cancer, pancreatic cancer, neuroblastoma, head and neck cancer, bladder cancer, kidney, brain, stomach or preferably rectum, prostate, breast, lung (especially non-small) Cell lung) or epithelium, in particular epidermal volume, for example cervical, cancer. Moreover, the diluent preparations of the present invention are also useful in the same way to treat the condition in which paclitaxel ^R is used. For certain tumors, epothilones have a more beneficial effect compared to paclitaxel. For certain tumors, for example certain types of lung tumors, for example the A549 lung, epothilone B has a more beneficial effect compared to paclitaxel.

In general, the diluent formulations of the present invention may be administered in therapeutically effective amounts for proliferative diseases that can be treated by the administration of epothilones such as epothilone A and / or epothilone B, in particular epothilone B. . Such proliferative diseases include any of the proliferative diseases mentioned above, in particular tumor diseases, and the response to a therapeutically effective amount preferably decreases proliferation, for example, decreases tumor growth or even (preferably) tumor regression. Or (most preferably) tumor removal. The exact dosage and duration depends on the nature of the epothilones such as epothilone A, epothilone B or mixtures thereof, the specific types of malignant proliferative cells specific to certain cancers, the severity of the condition, the rate of administration, and the patient's health and It may depend on the response to treatment.

In addition, pharmaceutical formulations of the present invention in a form suitable for parenteral administration, such as infusion solutions or reconstituted lyophilized compositions prepared by dilution of infusion concentrates, may be combined with other tumor therapies known to those skilled in the art, e. Or may be administered as part of a combination therapy comprising at least one other chemotherapeutic agent. The combined active agents can be administered simultaneously or sequentially, either of which is administered first. The dosage of the active agents of the combination therapy depends on the effectiveness and synergistic effect between the sites of action of each active agent and the agents used in the combination therapy.

Other chemotherapeutic agents may include any chemotherapeutic agent that may be used or may be used for the treatment of tumor diseases, in particular, such as chemotherapeutic agents derived from the following classes:

(A) alkylating agents, preferably crosslinking chemotherapeutic agents, preferably bis-alkylating agents,

(B) anticancer antibiotics, preferably doxorubicin (Adriamycin ^R , Rubex ^R )

(C) metabolic antagonists;

(D) plant alkaloids;

(E) hormones and antagonists;

(F) biological response modifiers, preferably lymphokines or interferons;

(G) protein tyrosine kinases and / or serine / threonine kinase inhibitors;

(H) antisense oligonucleotides or oligonucleotide derivatives; or

(I) other agent (s) with other or unknown mechanism of action, preferably taxanes, in particular taxotere ^R or most particularly paclitaxel (taxol ^R ).

Thus, the diluent preparations of the present invention may be useful as part of a single anticancer agent or a combination therapy for treating a variety of tumors.

The usefulness of any diluent formulation of the present invention is, for example, at the indicated epothilone doses that provide an equivalent serum concentration of epothilones; Standards using, for example, doses ranging from about 0.1 to 6 mg / m 2 for one week of treatment and about 0.3 to 18 mg / m 2 of epothilone for three week treatment of a 75 kilogram mammal, eg, 1.73 m 2 of adult It can be observed in clinical trials, or in standard animal models. For example, antitumor effects of single dose therapy are observed in human ovarian cancer models SKOV3 and U373 glioma models.

Increased bioavailability of epothilones administered in the diluent formulation form of the invention can be observed in standard animal tests and clinical trials, for example as described above. Naturally, the exact amount of epothilone and pharmaceutical agent administered can depend on a number of factors, such as the condition being treated, the exact epothilone, the desired duration of treatment and the rate of administration of the epothilone. For example, the amount of epothilone required and its rate of administration determine how long a particular epothilone concentration in plasma remains at an acceptable level of therapeutic effect, for example, as described above. And in vitro techniques.

Diluted formulations of the invention are conveniently about 0.1 to 100 mg / m 2, for example 0.2 to 100 mg / m 2 epothilone A and about 0.1 to 50 mg / m 2, for example 0.2 to 50 mg / m 2 epothilone B It can be administered intravenously in a dose. Preferably, for one week of treatment the dose is 0.1 to 6 mg / m 2, preferably 0.1 to 5, more preferably 0.1 to 3, even more preferably 0.1 to 1.7 mg / m 2, most preferably 0.1 to 1 mg / m 2; For three weeks of treatment the dose is from 0.3 to 18 mg / m 2, preferably from 0.3 to 15, more preferably from 0.3 to 12, even more preferably from 0.3 to 7.5 mg / m 2, most preferably from 1.0 to 3.0 mg / m 2. to be. These doses are preferably administered by intravenous administration to a human for 2 to 180 minutes, preferably 2 to 120 minutes, more preferably 5 to 30 minutes, most preferably 10 to 30 minutes, for example 30 minutes. Administered.

Preferably the concentration and dose strength is such that an effective dose level of about 0.5 to 15 mg / day, more preferably 1 to 10 mg / day, most preferably 2 to 8 mg / day can be obtained. Doses and blood levels received by intravenous administration can be accurately determined based on known in vivo and in vitro techniques.

In another aspect, the present invention provides a method of administering an epothilone to a subject in need of epothilone treatment comprising parenteral administration of the diluent formulation of the invention to a subject in need thereof. More specifically, such methods of administration of epothilone are prepared by (a) dilution of the pharmaceutical preparations according to the invention, for example in the form of infusion concentrates or lyophilized compositions, with an aqueous medium, followed by parenteral, for example intravenous, administration. And (b) administering such a solution to the subject.

In another aspect, the present invention provides the use of 에 epothilone in the manufacture of a medicament suitable for parenteral administration.

The invention is illustrated by the following examples, but is not intended to limit the scope of the invention. All percentages are weights / weight unless otherwise specified. Certain components of the pharmaceutical formulations are described in Fiedler, H.P., the contents of which are incorporated herein by reference. It may be further described in "Lexikon der Hilfsstoffe fur Pharmazie, Kosmetik und angrenzende Gebiete", Editio Cantor, D-7960 Aulendorf, 4th revised and expanded edition (1996).

<Example>

Example  One

Epothilone (B 15 mg or A 50 mg) is dissolved in 98-100% propylene glycol (1.0 mL). The solution is sterile filtered with a 0.22 micron pore size filter and filled in 1 ml ampoules. Filling ampoules are used for storage and shipping. The filling ampule is stable for at least 12 months at a temperature of 2 to 8 ° C. Prior to intravenous administration, the contents of the ampule are added to 250-1000 ml of a 5% glucose solution in water for injection. The intravenous solution so formed is stable for 8 hours at room temperature.

Example  2 to 7

The experiment of Example 1 is repeated using anhydrous and aqueous ethanol solvent systems and various polyethylene glycol solvent systems (Table 1).

TABLE 1

Example Solvent system 2 Anhydrous ethanol 3 Polyethylene Glycol 300 4 Polyethylene Glycol 400 5 50 to 100% ethanol 6 Polyethylene glycol 300: 90 to 100% 7 Polyethylene glycol 400: 90 to 100%

The injection solutions of Examples 2 to 7 are all stable within 8 hours at room temperature.

Example  8

Solubility in various solvent systems is summarized in Table 2. Unless indicated otherwise, all solubility results relate to T = 22 ° C.

TABLE 2

Solubility of Epothilone B in Various Solvent Systems

Solvent / Solvent-System Solubility [g / ℓ] ± 10% H ₂ O (pH 6.0) 0.16 (0.19, 4 ℃) Phosphate Buffer pH 7.4 0.16 (0.19, 4 ℃) H ₂ O, 0.9% glucose 0.16 H ₂ O, 5% glucose 0.16 H ₂ O, 15% Glycerin 0.19 H ₂ O, 5% poloxamer 188 0.23 EtOH / H ₂ O (v / v) 100/0 50/50 30/70 20/80 10/90 > 50 27 4.1 1.3 1.1 PEG 300 / H ₂ O (v / v) 100/0 50/50 70/30 12 10 2.5 PEG 400 / H ₂ O (v / v) 100/0 50/50 70/30 30 11 2.3 Propylene Glycol / H ₂ O (v / v) 100/0 70/30 50/50 26 10.4 1.6

The solubility of epothilone B in water at natural pH is about 160 mg / l, which results in significantly higher solubility in PEG / water, propylene glycol / water, or EtOH / water mixtures. In comparison, the previously reported water solubility of epothilone A is 940 mg / L and 700 mg / L for a mixture of epothilones A and B.

Example  9

The stability of the epothilone B-containing aqueous to non-aqueous polyethylene glycol infusion concentrates at different concentrations and at various temperatures is determined. Typically, a known amount of Epothilone B is dissolved in 1.0 ml of each of various solvent systems and each solution is sterile filtered to fill 1 ml white-glass vial with gray rubber stopper and gray flip-off cap. do. Table 3 lists the amount of degradation products formed over a period of up to seven months. The stability is analyzed by determining the formation of degradation products in each injection concentrate as a function of time and temperature. Each sample is analyzed by HPLC and the concentrate is diluted with an aqueous medium to prepare a sample. At high temperatures, for example 25 ° C., non-aqueous solvent systems comprising PEG generally exhibit higher stability than aqueous solvent systems comprising PEG.

TABLE 3

Proportion of degradation products in aqueous and non-aqueous PEG-containing injection concentrates

Solvent system Volume time 2-8 ℃ 25 ℃ PEG 400 1 mg / ml 3 months <0.1 0.2 PEG 400 / water 90:10 (w / w) 1 mg / ml 3 months <0.1 0.4 PEG 300 1 mg / ml 3 months <0.1 - PEG 300 / water 90:10 (w / w) 15 mg / ml 7 months <0.1 1.0 Ethanol / water 59:41 (w / w) 5 mg / ml First 1 month 5 months <0.1 0.3 0.3  -

Example  10

Epothilone B (5.0 mg) and mannitol (1500 mg) are dissolved in water for injection, and a 30 ml solution is prepared to prepare an aqueous solution. The solution is passed through a 0.22 micron pore size membrane filter before aseptically filling the glass vial, and the vials prepared for the drying process are aseptically plugged with sterile stoppers. The filled vial is then placed in a lyophilization chamber and cooled to about -40 ° C. The lyophilizer compactor is cooled to about -60 ° C and the chamber is evacuated to about 0.1 millitorr. Start the drying process by setting the chamber temperature to about 20 ° C. After about 20 hours of drying, the chamber pressure is increased to about 0.2 millitorr and the drying process is considered complete. Sterile air or nitrogen is introduced into the chamber aseptically to increase the chamber pressure to atmospheric pressure. The stopper then aseptically seals the vial to provide a sealed sterile seal. The sealed vial provides a single dose container of epothilone B that is reconstituted with 25 ml of water for injection immediately prior to administration. The dose is administered intravenously.

The lyophilized product has the desired properties required for the lyophilized composition according to the present invention.

Example  11 to 14

The method of Example 10 is carried out on the components described in the table below to form lyophilized products (Table 4).

TABLE 4

Example Epothilone B mg Hydroxypropyl-betacyclodextrin / mg Water for injection / ml 11 5.0 400 4.2 or less 12 5.0 500 5.0 or less 13 5.0 420 4.2 or less 14 5.0 420 8.5 or less

The lyophilized product formed according to Examples 11-14 has the desired properties required for the lyophilized composition according to the present invention.

Claims (7)

Lyophilized composition comprising Epothilone B and hydroxypropyl-beta-cyclodextrin. The lyophilized composition of claim 1, wherein epothilone B represents 0.1 to 1.5% by weight of total solids. The lyophilized composition of claim 2, wherein the hydroxypropyl-beta-cyclodextrin represents 90 to 99% by weight of the total solids. A reconstituted lyophilized composition comprising the lyophilized composition of any one of claims 1 to 3 in a pharmaceutically acceptable solvent. A method for preparing a medicament for parenteral administration using the lyophilized composition of any one of claims 1 to 3. The lyophilized composition of claim 1, wherein the freeze-dried composition comprises no surfactant having an HLB level of 10 or greater. A method for preparing a medicament for parenteral administration using the reconstituted lyophilized composition of claim 4.