HK1179859B - Oral dosage forms of bendamustine and therapeutic use thereof - Google Patents

Abstract

In the present invention there is provided a pharmaceutical composition for oral administration which comprises bendamustine or a pharmaceutically acceptable, ester, salt or solvate thereof as an active ingredient, and a pharmaceutically acceptable excipient and which shows a dissolution of the bendamustine of at least 60% in 20 minutes, 70% in 40 minutes and 80% in 60 minutes, as measured with a paddle apparatus at 50 rpm according to the European Pharmacopoeia in 500 ml of a dissolution medium at a pH of 1.5, and wherein the pharmaceutically acceptable excipient is either a pharmaceutically acceptable non-ionic surfactant, selected from the group consisting of a polyethoxylated castor oil or derivative thereof and a block copolymer of ethylene oxide and propylene oxide or a pharmaceutically acceptable saccharide selected from the group consisting of one or more of a monosaccharide, a disaccharide, an oligosaccharide, a cyclic oligosaccharide, a polysaccharide and a saccharide alcohol, wherein the ratio by weight of the active ingredient to the saccharide excipient(s) is in the range of 1:1-5. The invention further relates to the above pharmaceutical composition for use for the oral treatment of a medical condition which is selected from chronic lymphocytic leukemia, acute lymphocytic leukaemia, chronic myelocytic leukaemia, acute myelocytic leukaemia, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, breast cancer, ovarian cancer, small cell lung cancer and non-small cell lung cancer. The invention moreover relates to the above pharmaceutical composition for the above use wherein the dosage regimen comprises at least the administration of a dose of 100 to 600 mg/m2/per person of bendamustine on day 1 and day 2, optionally a dose of 50 to 150 mg/m2 i.v. or orally of a corticosteroid on days 1 to 5, and optionally a suitable dose of a further active agent selected from the group consisting of an antibody specific for CD20, an anthracyclin derivative, a vinca alkaloid or a platin derivative; and the repetition of said dosage regimen 4 to 15 times after intervals of two to four weeks.

Description

Oral dosage forms of bendamustine and therapeutic uses thereof

Technical Field

The present invention relates to oral dosage forms comprising bendamustine, or a pharmaceutically acceptable ester, salt, or solvate thereof, and therapeutic uses thereof.

Background

Bendamustine (4- [5- [ bis (2-chloroethyl) amino ] -1-methylbenzimidazol-2-yl ] butanoic acid, a nitrogen mustard) is an alkylating agent with bifunctional alkylating activity. Which corresponds to the following structural formula (I):

bendamustine appears to be free of any cross-resistance to other alkylating agents, which has advantages in chemotherapy for patients who have already been treated with alkylating agents.

Bendamustine was originally synthesized in the german democratic republic of morale (GDR). Under the trade name of 1971 to 1992Bendamustine hydrochloride is an active ingredient in commercial products sold. Since then, bendamustine hydrochloride has been under the trade name in germanyAre marketed and widely used for the treatment of chronic lymphocytic leukemia, non-hodgkin's lymphoma and multiple myeloma.

The commercial product comprises bendamustine hydrochloride lyophilized powder which is reconstituted with water for injection to produce a concentrate. This was then diluted with 0.9% aqueous sodium chloride to give the final solution for infusion. The final solution is administered to the patient by intravenous infusion over a period of about 30 to 60 minutes.

Hydrolysis of the di-2-chloroethylamino group of bendamustine in water results in decreased potency and the formation of impurities (B.Maas et al (1994), Pharmazie49: 775-777). It must therefore be administered immediately after reconstitution of the lyophilized powder (usually in a hospital or at least under medical supervision). Furthermore, it has been reported that the reconstitution process is difficult. This process may take more than 30 minutes. In addition, reconstitution in a 2-step process is cumbersome and time consuming for medical professionals responsible for reconstituting products.

Preiss et al (1985) (Pharmazie 40: 782-784) compared the pharmacokinetics of bendamustine hydrochloride in the plasma of 7 patients following intravenous and oral administration at doses of 4.2mg/kg to 5.5mg/kg, respectively. From commercial productsThe prepared intravenous infusion was administered within 3 minutes, while an equivalent dose of the oral drug was administered in the form of a capsule containing 25mg bendamustine hydrochloride. The number of capsules administered to patients varies from 10 to 14, i.e. an absolute oral dose of 250mg to 350 mg. After oral administration, maximum plasma levels were detectable within 1 hour. The average oral bioavailability was calculated to be 57%, ranging from 25% to 94%, showing large inter-individual differences (% CV = 44%). Similar studies with larger inter-individual differences (25% to 121%) were reported by Preiss et al in later literature (Z.Klin. Med.44(1989): 125-129).

Weber (1991) (Pharmazie 46(8): 589-591) studied the bioavailability of bendamustine hydrochloride in B6D2F 1-mice and found that the drug was not completely absorbed from the gastrointestinal tract, which resulted in a bioavailability of only about 40%.

Patent document US2006/0128777a1 describes a method of treating cancer which is generally characterized by anti-dead cells and a bendamustine-containing composition. Among these compositions are oral dosage forms in the form of capsules, tablets, pills, powders or granules, in which the active compound may be mixed with at least one inert excipient (for example sucrose, lactose or starch). However, it does not exemplify a specific composition.

Bendamustine hydrochloride is only sparingly soluble in water at pH 2.0 and is soluble in a range of organic solvents in trace or minimal amounts. However, it has been observed to show good solubility in ethanol and methanol. Therefore, not surprisingly, oral bendamustine compositions, as studied by Preiss et al and Weber, produce relatively poor bioavailability results and large inter-individual variation.

In view of the stability problems associated with commercially available intravenous formulations once reconstituted with water, and in order to improve patient compliance, there has long been a need for a stable dosage form containing bendamustine which is easy to administer to patients and which provides good bioavailability without large inter-and intra-individual variations. There is also a need for a pharmaceutical composition that allows complete absorption, or at least a substantial degree of absorption, of bendamustine in the stomach, thereby avoiding or reducing degradation of bendamustine in the small or large intestine.

Disclosure of Invention

The present inventors have conducted detailed studies in order to solve the above problems. They finally succeeded in obtaining the stable pharmaceutical composition of the present invention. These compositions are suitable for oral administration and comprise: bendamustine or a pharmaceutically acceptable ester, salt or solvate thereof as active ingredient, together with at least one pharmaceutically acceptable excipient, which composition, in addition to having good stability, has good dissolution behavior in acidic media, good bioavailability and therapeutically acceptable inter-and intra-individual differences.

Drawings

Figure 1 shows the mean plasma concentration versus time curves obtained after administration of bendamustine hydrochloride to puppies in the form of a prior art capsule (reference example 1) and in the form of a liquid-filled hard capsule formulation of example 2. Figure 1 clearly shows that the liquid-filled hard capsule formulation provides a higher maximum concentration of bendamustine compared to the prior art reference capsule formulation.

FIG. 2 shows the preparation for intravenous administration (in Germany)Sales) and the mean plasma concentration versus time profile obtained after administering bendamustine hydrochloride to a cancer patient in the form of the liquid-filled hard capsule formulation of example 2.

Figure 3 shows the mean plasma concentrations (tablet versus capsule) versus time curves obtained after administration of bendamustine hydrochloride to puppies in the form of prior art capsules and in the form of tablet formulations of examples 15 to 17 (tablets 1 to 3) and example 18 (formulation 3) (tablet 4). Figure 3 clearly shows that the tablet formulation is able to provide a higher maximum concentration of bendamustine compared to the prior art capsules.

Figure 4 shows a flow chart of a wet granulation manufacturing run.

Detailed Description

The present invention relates to a pharmaceutical composition for oral administration comprising: bendamustine or a pharmaceutically acceptable ester, salt or solvate thereof as active ingredient, and a pharmaceutically acceptable excipient, and which pharmaceutical composition exhibits such bendamustine dissolution behaviour when measured according to the european pharmacopoeia in 500ml of dissolution medium at a pH of 1.5 using a paddle apparatus rotating at 50 rpm: at least 60% after 20 minutes, at least 70% after 40 minutes and at least 80% after 60 minutes and wherein the pharmaceutically acceptable excipient is a pharmaceutically acceptable non-ionic surfactant selected from the group consisting of polyethoxylated castor oil or a derivative thereof and a block copolymer of ethylene oxide and propylene oxide or a pharmaceutically acceptable sugar selected from the group consisting of one or more of a monosaccharide, a disaccharide, an oligosaccharide, a cyclic oligosaccharide, a polysaccharide and a sugar alcohol, wherein the weight ratio of the active ingredient to the sugar excipient is in the range of 1 (1-5).

In a first embodiment of the invention, the invention relates to a pharmaceutical composition for oral administration comprising: bendamustine or a pharmaceutically acceptable ester, salt or solvate thereof as an active ingredient, and a pharmaceutically acceptable excipient which is a nonionic surfactant selected from the group consisting of polyethoxylated castor oil or a derivative thereof, and a block copolymer of ethylene oxide and propylene oxide.

One embodiment of the first embodiment of the present invention is a pharmaceutical composition comprising: bendamustine or a pharmaceutically acceptable ester, salt or solvate thereof, and a pharmaceutically acceptable excipient which is a non-ionic surfactant selected from the group consisting of polyethoxylated castor oil or a derivative thereof, and a block copolymer of ethylene oxide and propylene oxide, wherein the composition is suitable for oral administration by filling into a gelatin hard capsule.

Another embodiment of the first embodiment of the present invention is a pharmaceutical composition for oral administration in the form of a solid dosage form (hard gelatin capsule), comprising: bendamustine or a pharmaceutically acceptable ester, salt or solvate thereof as active ingredient, and a pharmaceutically acceptable excipient selected from the group consisting of polyethoxylated castor oil or a derivative thereof, and a block copolymer of ethylene oxide and propylene oxide, and preferably selected from the group consisting of macrogol glycerol hydroxystearate, polyoxyethylene-35-castor oil (polyoxyl-35-castor oil), and an ethylene oxide/propylene oxide block copolymer(s) ((r))L44NF or124) Wherein the use of said specific non-ionic surfactant results in such dissolution behavior: bendamustine is dissolved after 20 minutes at least 60%, after 40 minutes at least 70% and after 60 minutes at least 80%, as measured according to the european pharmacopoeia in 500ml of dissolution medium at pH 1.5 using a paddle apparatus with a rotation speed of 50rpm, and preferably it results in a dissolution behavior: bendamustine dissolves at least 60% after 10 minutes, at least 70% after 20 minutes, and at least 80% after 30 minutes.

A preferred embodiment of said first embodiment is a pharmaceutical composition for oral administration in a solid dosage form (hard gelatin capsule) comprising bendamustine hydrochloride and macrogolglyceride hydroxystearate as a pharmaceutically acceptable excipient, wherein the use of said specific non-ionic surfactant results in such dissolution behavior: bendamustine dissolves at least 60% after 10 minutes, at least 70% after 20 minutes and at least 80% after 30 minutes as measured according to the european pharmacopoeia in 500ml of dissolution medium having a pH of 1.5 using a paddle apparatus rotating at 50 rpm.

In a second embodiment of the present invention, the present invention relates to a pharmaceutical composition comprising: bendamustine or a pharmaceutically acceptable ester, salt or solvate thereof as an active ingredient, and at least one pharmaceutically acceptable excipient selected from the group consisting of monosaccharides, disaccharides, oligosaccharides, cyclic oligosaccharides, polysaccharides and sugar alcohols. Preferably, the weight ratio between the active ingredient and the excipient is in the range of 1:1 to 1:5, preferably in the range of 1:2 to 1:5, more preferably a ratio selected from 1:5 and 1: 2.

In one embodiment of the second embodiment of the present invention, the present invention relates to a solid dosage form pharmaceutical composition for oral administration, the composition comprising: bendamustine or a pharmaceutically acceptable ester, salt or solvate thereof as an active ingredient, and at least one pharmaceutically acceptable excipient which is a pharmaceutically acceptable sugar selected from the group consisting of one or more of a monosaccharide, a disaccharide, an oligosaccharide, a cyclic oligosaccharide, a polysaccharide and a sugar alcohol, wherein the weight ratio of the active ingredient to the excipient is in the range of 1:1.

In a further embodiment of the second embodiment of the present invention, the present invention relates to a solid dosage form pharmaceutical composition suitable for oral administration, the composition comprising: bendamustine or a pharmaceutically acceptable ester, salt or solvate thereof as active ingredient, and at least one pharmaceutically acceptable excipient which is a pharmaceutically acceptable sugar selected from the group consisting of one or more of a monosaccharide, a disaccharide, an oligosaccharide, a cyclic oligosaccharide, a polysaccharide and a sugar alcohol, wherein the weight ratio of the active ingredient to the sugar excipient is in the range of 1:2 to 1:5, and the composition exhibits bendamustine dissolution behavior as follows: bendamustine dissolves at least 60% within 20 minutes, at least 70% within 40 minutes and at least 80% within 60 minutes as measured according to the european pharmacopoeia in 500ml of dissolution medium at pH 1.5 using a paddle apparatus rotating at 50 rpm.

Within the scope of the second embodiment above, a further preferred embodiment is a pharmaceutical composition wherein the pharmaceutically acceptable sugar is selected from the group consisting of one or more of mono-, di-and oligosaccharides, wherein the weight ratio of the active ingredient to the sugar excipient is in the range of 1:2 to 1:5, and wherein the composition exhibits bendamustine dissolution behavior as follows: bendamustine dissolves at least 60% within 20 minutes, at least 70% within 40 minutes and at least 80% within 60 minutes as measured according to the european pharmacopoeia in 500ml of dissolution medium at pH 1.5 using a paddle apparatus rotating at 50 rpm.

The present invention is based on the unexpected discovery that: stable bendamustine compositions with specific and good dissolution behavior may be obtained by incorporating certain nonionic surfactants or certain sugars or sugar alcohols into the pharmaceutical composition. It has been found that particularly good behaviour in terms of stability and degradation products, dissolution profile, bioavailability and reduced differences in bioavailability is obtained if the following pharmaceutically acceptable nonionic surfactants are used as excipients in pharmaceutical compositions comprising bendamustine or a pharmaceutically acceptable ester, salt or solvate thereof as active ingredient, selected from the group consisting of polyethoxylated castor oil or a derivative thereof, and block copolymers of ethylene oxide and propylene oxide, and preferably selected from the group consisting of polyglycolized castor oil or a derivative thereof, polyoxyethylene-35-castor oil and ethylene oxide/propylene oxide block copolymers (polyoxyethylenegylcetalated glyceryl stearate, polyoxyethylene35-castor oil and ethylene oxide/propylene oxide block copolymers: (a)L44NF or124) Group (d) of (a). The incorporation of the above-mentioned nonionic surfactants in bendamustine-containing compositions results in such dissolution behavior: bendamustine is dissolved after 20 minutes at least 60%, after 40 minutes at least 70% and after 60 minutes at least 80%, as measured according to the european pharmacopoeia in 500ml of dissolution medium at pH 1.5 using a paddle apparatus with a rotation speed of 50rpm, and which preferably results in a dissolution behavior of: bendamustine dissolves at least 60% after 10 minutes, at least 70% after 20 minutes, and at least 80% after 30 minutes.

It has further been found that particularly good behaviour in terms of stability, tabletting properties, dissolution and impurity formation is achieved if a pharmaceutically acceptable sugar selected from the group consisting of one or more monosaccharides, disaccharides, oligosaccharides, cyclic oligosaccharides, polysaccharides and sugar alcohols, and preferably from the group consisting of one or more monosaccharides, disaccharides and oligosaccharides is used as excipient in a pharmaceutical composition comprising bendamustine or a pharmaceutically acceptable ester, salt or solvate thereof as active ingredient. The above sugars allow the composition to exhibit the following bendamustine dissolution behavior: bendamustine dissolves at least 60% within 20 minutes, at least 70% within 40 minutes and at least 80% within 60 minutes as measured according to the european pharmacopoeia in 500ml of dissolution medium at pH 1.5 using a paddle apparatus rotating at 50 rpm.

Within the above scope of the second embodiment of the present invention, any combination of one or more of monosaccharides, disaccharides, oligosaccharides, cyclic oligosaccharides, polysaccharides and sugar alcohols may be used.

In particular, it has been found that specific sugars are associated with a particularly good behaviour of the pharmaceutical composition in terms of stability and dissolution. The sugar in the composition of the second embodiment of the invention is preferably anhydrous dextrose, dextrose monohydrate, lactitol monohydrate, trehalose, sorbitol, erythritol, maltose monohydrate, mannitol, lactose anhydrous, lactose monohydrate, maltitol, xylitol, sucrose 97% + maltodextrin 3%, beta-cyclodextrin, D-raffinose pentahydrate, D-melezitose monohydrate and microcrystalline cellulose. The pharmaceutical composition of the present invention exhibits good tabletting characteristics, rapid dissolution behavior and pharmaceutically suitable stability.

The above sugars constitute a preferred embodiment of the second embodiment of the present invention and any combination thereof may be used. Preferably, the ratio of the active ingredient to the above sugar is in the range of 1:1 to 1:5, preferably in the range of 1:2 to 1:5, and more preferably is a ratio selected from 1:5 and 1: 2.

A further preferred embodiment of the second embodiment of the present invention is a pharmaceutical composition in solid dosage form for oral administration, the composition comprising: bendamustine or a pharmaceutically acceptable ester, salt or solvate thereof as an active ingredient, and at least one pharmaceutically acceptable excipient selected from the group consisting of anhydrous dextrose, dextrose monohydrate, lactitol monohydrate, trehalose, sorbitol, erythritol, maltose monohydrate, mannitol, anhydrous lactose, lactose monohydrate, maltitol, xylitol, sucrose 97% + maltodextrin 3%, β -cyclodextrin, D-raffinose pentahydrate, D-melezitose monohydrate and microcrystalline cellulose, said composition exhibiting: bendamustine dissolves at least 60% within 10 minutes, at least 70% within 20 minutes, and at least 80% within 30 minutes.

Particularly preferred sugars are mannitol, maltitol, erythritol, xylitol, lactose, sucrose, glucose (glucose), sorbitol, maltose, trehalose, lactitol and dextrose (dextrose) (anhydrous or monohydrate), and the weight ratio of the active ingredient to the sugar is preferably in the range of 1:2 to 1: 5. The invention also includes combinations of two or more sugars within the scope of the above sugars.

The person skilled in the art is fully enabled to select suitable combinations among the above mentioned sugar excipients and to obtain compositions showing the following bendamustine dissolution behaviour: bendamustine is at least 60% dissolved in 20 minutes, at least 70% dissolved in 40 minutes and at least 80% dissolved in 60 minutes, measured according to the european pharmacopoeia in 500ml of dissolution medium at pH 1.5 using a paddle apparatus rotating at 50 rpm.

In a preferred embodiment, the composition is in the form of a tablet, granule, or pill.

A preferred dosage form is a tablet, and is preferably an immediate release tablet (i.e. a tablet which releases the active ingredient very quickly after the tablet has been placed in an aqueous medium, preferably an acidic medium. The term tablet also includes fast disintegrating tablets, among which dispersible tablets and effervescent tablets.

The most common tablet preparation methods are direct compression, dry granulation and wet granulation. Direct compression involves compression molding a mixture containing the active ingredient and excipients on a tablet press (l.lachman et al, the orthoandpracticephof industril pharmacy, third edition, 1986). In order to prepare tablets having a uniform content of active ingredient, the mixture to be compression-molded must have both good flowability and good compression moldability. It is not always possible to obtain good flowability by adding suitable excipients to the mixture, such as lubricants, anti-adherents and flow promoters. The mixture is often granulated before compression molding.

Granulation is a process in which a powdery mixture is formed into aggregates (called granules) having an approximately spherical or regular shape. This process can be achieved by dry granulation and wet granulation. Granulation is also used to convert powder mixtures with low cohesion into aggregates which when compression molded give tablets with good cohesive properties.

In the case of rapidly disintegrating tablets, the active ingredient, optionally mixed with one or more excipients, is advantageously provided with a coating to mask the taste of the active ingredient and/or to protect the active ingredient from possible harmful effects caused by light and/or moisture, and in the case of bendamustine, to protect the oral mucosa from harmful effects caused by the active compound. For this purpose, the granules are preferably prepared and processed as further described below.

The expression "particle" refers to an aggregate of particles, sometimes also referred to as a pellet. Granules are generally prepared by compaction and/or compression moulding techniques (dry granulation); or by wet granulation with a liquid in which a wet granulation binder is optionally dissolved (Remington, pharmaceutical sciences, 18 th edition, 1990, page 1641). Wet granulation techniques also include extrusion techniques. The term granulate therefore also includes pellets, prills, and extrudates, wherein a preferred example of a granulate is a pellet.

Pellets can be described as small granules having a certain density and a diameter of about 1.0 to 1.6mm, which are prepared by applying a pharmaceutical process of extrusion spheronization to a powdered mixture.

The active ingredient, optionally mixed with one or more excipients, may advantageously be provided with a coating to mask the taste of the active ingredient and/or to protect the active ingredient from possible harmful effects caused by light and/or moisture and/or to protect the oral mucosa from harmful effects caused by the active ingredient.

The pellets are small and round solid dosage forms prepared by adding the active ingredient to a soft triglyceride mixture. The mixture was rolled into a long strand, which was then cut into sections and rolled (j.t. carstensen, pharmaceutical printersofsoliddosages, 1993, technomic publishing company, page 63).

The dosage forms of the present invention are preferably prepared by dry compaction techniques. Suitable techniques are described, for example, in Remington's pharmaceutical sciences, 18 th edition, 1990, page 1644. They include dry granulation, rolling and direct compression. When tablets are prepared using these techniques, direct compression is more advantageous.

The dosage form according to the invention preferably has a coating. The coating has different purposes: can be used to mask the taste of the active ingredients used in the composition while protecting said active ingredients from possible harmful effects caused by light and/or moisture, such as oxidation, degradation, etc. In addition, the coating layer may protect the oral mucosa of the subject from damage caused by the active ingredient.

The coating layer may be applied to the dosage form by techniques well known in the art, such as spraying and microencapsulation. For tablets, they may be in the form of film coatings, sugar coatings or press coatings. Preferably, a film coating process is used (Remington, pharmaceutical sciences, 18 th edition, 1990, page 1666). In the case of rapidly disintegrating tablets coated as required by the active ingredient, the individual granules may be suitably coated before compression molding into tablets.

The expression "pharmaceutically acceptable ester thereof" refers to any pharmaceutically acceptable ester of bendamustine, such as the ester of bendamustine with an alkyl alcohol and the ester with a sugar alcohol. An example of an alkyl alcohol is C_1-6Alkyl alcohols such as methanol, ethanol, propanol, isopropanol, butanol and tert-butanol. Examples of sugar alcohols are mannitol, maltitol, sorbitol, erythritol, ethylene glycol, glycerol, arabitol, xylitol and lactitol. Preferred examples of bendamustine esters are ethyl esters, isopropyl esters, mannitol esters, and sorbitol esters, most preferably bendamustine ethyl ester.

The expression "pharmaceutically acceptable salt thereof" refers to any pharmaceutically acceptable salt of bendamustine that is administered to a patient (directly or indirectly) to provide bendamustine. The term also includes pharmaceutically acceptable salts of bendamustine esters. However, pharmaceutically unacceptable salts are also considered to be within the scope of the present invention, as these compounds may be used to prepare pharmaceutically acceptable salts. For example, pharmaceutically acceptable salts of bendamustine are synthesized from the corresponding compounds containing acid or base groups by conventional chemical methods. These salts are typically prepared, for example, by reacting the free acid or free base forms of these compounds with the corresponding base or acid in stoichiometric amounts in water or in an organic solvent, or in a mixture of the two. Non-aqueous media such as ether, ethyl acetate, isopropanol or acetonitrile are generally preferred. Examples of acids useful in forming pharmaceutically acceptable salts of bendamustine include: inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, and phosphoric acid; and organic acids such as acetic acid, maleic acid, fumaric acid, citric acid, oxalic acid, succinic acid, tartaric acid, malic acid, lactic acid, methanesulfonic acid, and p-toluenesulfonic acid. Pharmaceutically acceptable salts of bendamustine may be derived from inorganic or organic bases to yield ammonium salts, alkali metal salts (lithium, sodium, potassium, etc.), alkaline earth metal salts such as calcium or magnesium, aluminum salts, lower alkyl amine salts such as methylamine salts or ethylamine salts, lower alkyl diamine salts such as ethylenediamine salts, ethanolamine salts, N-dialkylene ethanolamine salts, triethanolamine salts, and glucamine salts, and basic salts of amino acids. Especially preferred are the acid salts prepared from hydrochloric, hydrobromic and hydroiodic acid, with the hydrochloride salt being the most preferred pharmaceutically acceptable salt of bendamustine. The pharmaceutically acceptable salts are prepared by conventional techniques well known in the art.

The expression "pharmaceutically acceptable solvate thereof" refers to any pharmaceutically acceptable solvate that is administered to a patient (directly or indirectly) to provide bendamustine. The term also includes pharmaceutically acceptable solvates of bendamustine esters. The solvate is preferably a hydrate, a solvate with an alcohol such as methanol, ethanol, propanol or isopropanol, a solvate with an ester such as ethyl acetate, a solvate with an ether such as methyl ether, ethyl ether or THF (tetrahydrofuran), or a solvate with DMF (dimethylformamide), and among them, a hydrate or a solvate with an alcohol such as ethanol is more preferable. The solvent constituting the solvate is preferably a pharmaceutically acceptable solvent.

It is especially preferred that the active ingredient in the composition of the present invention is bendamustine or a pharmaceutically acceptable salt thereof. Most preferably, the active ingredient is bendamustine hydrochloride.

The dosage of the active ingredient in the pharmaceutical composition may be determined by the skilled personThe member is selected according to the condition, sex, weight, and body surface area (m) of the patient²(ii) a Average about 2m²Person) or age, in particular on the basis of the body weight and body surface area of the patient. Preferably, the daily dose of the active ingredient is from about 50mg to about 1000mg, preferably from about 100mg to about 500mg, more preferably from about 200mg to about 400mg, and most preferably about 280 mg. The daily dose may be administered in a single dose or in multiple doses, for example twice or three times daily, most preferably in a single daily dose. The daily dose may be administered once a week or several times a week. The minimum oral single dose is 50 mg. The above dosages relate to bendamustine and, where relevant to pharmaceutically acceptable esters, salts or solvates thereof, the above dosages can be readily recalculated. The dose may be expressed in absolute amounts (mg), but in oncology, the dose is usually expressed in mg/m taking into account the body surface area of the patient²。

The Maximum Tolerated Dose (MTD) and effective dose of bendamustine depends on the cumulative amount administered per cycle. Cumulative amounts of MTD per cycle up to 1000mg were measured according to the reproducible ba (reproducibleba) of bendamustine. The lower limit of the effective cumulative dose per cycle is between 350mg and 500 mg. Thus, a cumulative dose per cycle of 350mg to 1000mg needs to be administered orally. Preferred oral cumulative doses per cycle are 500mg to 700 mg. Bendamustine may be provided in effective single doses of from 50mg to 900 mg. The preferred range for an oral single dose is 200mg to 300 mg.

The maximum tolerated dose (cumulative) for one cycle (3-4 week cycle) was about 1000mg bendamustine. In susceptible/compromised patients, the cumulative dose over one cycle (3-4 weeks) is about 350-500mg bendamustine, preferably about 365mg over 4 weeks.

Possible and preferred oral dosage regimens are:

-a low maintenance dose of 200mg to 300mg on day 1 and day 2, optionally followed by 50mg once a day.

From day 1 to day 14 (including day 14), 50mg bendamustine per day.

-about 150mg bendamustine, once a week for 3 weeks.

Typically, treatment with bendamustine is effective over a treatment cycle in which bendamustine and optionally additional agents are administered for 1 to 5 days, followed by repetition of the treatment after 2 to 4 weeks of discontinuation. This cycle of treatment is repeated continuously until the corresponding condition to be treated is improved. Basically, the number of repetitions is determined by the physician. Typically, the treatment cycle is repeated 4 to 15 times, preferably 4 to 12 times, more preferably 4 to 6 times.

The following effective (intravenous administration) and preferred oral dosing regimens for the specific indications within the scope of the invention are given:

monotherapy for chronic lymphocytic leukemia:

the composition is administered at a dose of 100mg/m on day 1 and day 2²Bendamustine hydrochloride at body surface area; once every 4 weeks (intravenous administration).

Oral administration: 145mg/m²Or 261mg (1.8 m)²): 100-200mg/m per day²Or 150-350 mg.

Monotherapy rituximab refractory indolent non-hodgkin lymphoma:

at day 1 and day 2, 120mg/m²Bendamustine hydrochloride at body surface area; once every 3 weeks (intravenous administration).

Oral administration: 174mg/m²Or 313mg (1.8 m)²): 100-250mg/m per day²Or 150-400 mg.

Preferably, bendamustine is used in combination with vincristine and prednisone in the first-line treatment of non-hodgkin's lymphoma.

Multiple myeloma:

at day 1 and day 2, 120-150mg/m²Body surface area bendamustine hydrochlorideStatin (intravenous administration); administered by intravenous injection or oral administration at 60mg/m on days 1 to 4²Prednisone on body surface area, once every 4 weeks.

Oral administration: 174-217mg/m²Or 313-391mg (1.8 m)²): 100-250mg/m per day²Or 150-400 mg.

Combination therapy for first line treatment of patients with Follicular Lymphoma (FL), indolent lymphoma, and Mantle Cell Lymphoma (MCL):

375mg/m rituximab²(day 1) + bendamustine 90mg/m²(day 1 + day 2), once every 28 days (intravenous administration).

Oral administration: 130mg/m²Or 235mg (1.8 m)²): 100-200mg/m per day²Or 150-350 mg.

Accordingly, the present invention relates to a pharmaceutical composition as defined above for the oral treatment of a medical condition selected from chronic lymphocytic leukemia, acute lymphocytic leukemia, chronic myelocytic leukemia, acute myelocytic leukemia, hodgkin's disease, non-hodgkin's lymphoma, multiple lymphoma, breast cancer, ovarian cancer, small cell lung cancer and non-small cell lung cancer, wherein the dosing regimen comprises: the dose is 100mg/m on at least day 1 and day 2²One person to 600mg/m²Bendamustine/human, optionally administered by intravenous injection or orally on days 1 to 5 at 50mg/m²To 150mg/m²And optionally administering a suitable dose of an additional active agent selected from the group consisting of a CD 20-specific antibody, an anthracycline derivative, vinblastine or a platinum derivative; and after 2-4 weeks intervals, the dosing regimen is repeated 4 to 15 times. Furthermore, the present invention relates to the pharmaceutical composition as defined above for the use as defined above, wherein the active ingredient bendamustine is administered in a dosage regimen selected from the group consisting of: 200-300mg was administered on days 1 and 2; optionally followed once a day by a maintenance dose of 50 mg; from day 1 to day 14 (including day 14)) 50mg per day; or 150mg once a week for 3 weeks.

The present invention also relates to a pharmaceutical composition as defined above for use as defined above, wherein the patient is a human suffering from non-hodgkin's lymphoma and the dosing regimen comprises: the total amount of active ingredient bendamustine was 200 mg/person/day on days 1 to 5, 2mg vincristine was administered intravenously on day 1, and 100mg/m was administered intravenously on days 1 to 5²Until non-hodgkin lymphoma is ameliorated, the treatment is repeated every three weeks.

The present invention also relates to a pharmaceutical composition as defined above for use as defined above, wherein the patient is a human suffering from multiple myeloma and the dosing regimen comprises: the dosage on the 1 st day and the 2 nd day is 100-250mg/m²Body surface area, preferably 174mg/m²Bendamustine hydrochloride at body surface area, and administered 60mg/m intravenously or orally from day 1 to day 4²Until the multiple myeloma is improved, the treatment is repeated every four weeks.

The present invention also relates to a pharmaceutical composition as defined above for use as defined above, wherein the patient is a human suffering from chronic lymphocytic leukemia and the dosing regimen comprises: the dosage on the 1 st day and the 2 nd day is 100-200mg/m²Body surface area, preferably 145mg/m²Bendamustine hydrochloride at body surface area, and administered 60mg/m intravenously or orally from day 1 to day 4²Said treatment is repeated every four weeks until said chronic lymphocytic leukemia is ameliorated.

The present invention further relates to a pharmaceutical composition as defined above for use as defined above, wherein the patient is a human suffering from follicular lymphoma, indolent lymphoma or mantle cell lymphoma, and the dosing regimen comprises: the dosage is 375mg/m on day 1 and day 2²And orally additionally administered 100mg/m on days 1 and 2²To 200mg/m²Preferably 130mg/m²The bendamustine of (c) was repeated every 28 days until the corresponding lymphoma was ameliorated.

The dosage form may comprise a single daily dose amount or a partial amount thereof. Preferably, the dosage form of the present invention comprises from about 10mg to about 1000mg, preferably from about 25mg to about 600mg, more preferably from about 50mg to about 200mg, and most preferably about 50mg or about 100mg of the active ingredient.

As used herein, the term "nonionic surfactant" refers to an amphiphilic compound having a polar hydrophilic group and a non-polar lipophilic group or chain, and wherein the hydrophilic and lipophilic properties of the compound are characterized by the so-called hydrophilic-lipophilic balance (HLB) value. Preferably, the nonionic surfactant used to prepare the compositions of the present invention has an HLB value between 10 and 20, more preferably between 12 and 18. The nonionic surfactant also has a melting point, flow point or melting range (meltrange) between 5 ℃ and body temperature (37 ℃), preferably just below room temperature (20 ℃) and body temperature. The material may be liquid or semi-solid at room temperature. The amphiphilic material is a carrier for the bendamustine active ingredient, which may be present in dissolved form, suspended form, or partially dissolved partially suspended form.

The nonionic surfactant advantageously used in the preparation of the compositions of the present invention is selected from the group consisting of polyethoxylated castor oil or derivatives thereof and ethylene oxide/propylene oxide block copolymers, provided that these materials have the aforementioned HLB and melting points, flow points or melting ranges.

In one embodiment, the nonionic surfactant is a polyethoxylated castor oil. An example of polyethoxylated castor oil is known under the trade name castor oilAnd (5) selling. Having various purities and viscositiesProduct has been madeAre ready for use in the present invention. In particular, polyglycolized glyceride hydroxystearate(s) ((II))RH 40) and polyoxyethylene-35-castor oil (R: (R)EL orELP). It is known thatELP andEL as a nonionic solubilizer and emulsifier was prepared by reacting castor oil with ethylene oxide in a molar ratio of 1: 35. They have an HLB value of 12 to 24 and a melting point of 26 ℃. Depending on the ambient temperature, these products can be characterized as semi-solid or medium viscosity liquids. Glyceryl polyethylene glycol hydroxystearate (or polyethylene glycol glyceryl monostearate)RH40 commercially available) is a semi-solid material at 25 c and has a viscosity in the range of 20cps to 40cps (as a 30% aqueous solution) at the same temperature. Polyglycolized hydroxystearate is known as a nonionic solubilizer and emulsifier. It is prepared by reacting castor oil with ethylene oxide in a molar ratio of 1: 45. It has an HLB value ranging from 14 to 16 and a melting range of 20-28 ℃. Tests have shown that polyglycolized glyceride hydroxystearate itself can be advantageously used in the preparation of the compositions of the present invention.

The block copolymer is composed of ethylene oxide block and propylene oxideAn alkane block. The ethylene oxide units are hydrophilic, while the propylene oxide units are lipophilic. The varying number of hydrophilic ethylene oxide units and lipophilic propylene oxide units results in copolymers having different molecular weights and different hydrophilic-lipophilic balance (HLB) values. Examples of propylene oxide ("PEO") -polypropylene oxide ("PPO") block copolymers having HLB values and melting points or flow points or melting ranges consistent with the requirements for preparing the compositions of the present invention include the types that are commercially availableL35、L44、L64、P85 andP105。l44 or124, but not exclusively68 or188 and127 or407。L44 is a preferred nonionic surfactant.

In addition to the hydroxystearic acid macrogolglycerides, the above nonionic surfactants are liquids whose viscosity values may be too low to avoid precipitation of bendamustine hydrochloride. Another problem to be solved is to find an excipient or combination of excipients that allows the overall viscosity of the mixture to be high enough to avoid separation of bendamustine chloride upon addition to the mixture.

Thus, the liquid nonionic surfactant-containing composition of the present invention also advantageously comprises a viscosity modifier. Suitable viscosity-improving agents include: powders, e.g. colloidal silica (trade name)Commercially available); or semi-solid waxy materials, e.g. lauroyl macrogolglycerides (trade name)44/14 commercially available). The amount of powder or semi-solid material added to the liquid nonionic surfactant depends on the viscosity of the liquid nonionic surfactant. Different concentrations have been tested to find the appropriate minimum amount of viscosity modifier added to avoid visible active ingredient precipitation. The colloidal silica is typically added in a relative concentration of about 1% to about 8%, but preferably as low as 1.7% or 2.0%, in order not to adversely affect the dissolution characteristics of the active ingredient. Typically the relative concentration of lauroyl macrogolglycerides is from 5% to 50%, and preferably about 10% and about 45%.

A preferred composition of the invention is disclosed in example 4 and comprises bendamustine hydrochloride in combination with:

-polyethylene glycol glycerol hydroxystearate;

ethylene oxide/propylene oxide block copolymers (C)L44NF or124) Optionally with colloidal silicon dioxide or lauroyl macrogolglycerides (44/14) combinations, and

polyoxyethylene-35-castor oil, optionally with lauroyl macrogolglycerides (44/14) are combined.

Advantageously, the pharmaceutical composition of the invention is filled into capsules, which may allow easy administration to the patient.

Two types of capsules are generally used and can be classified according to the nature and flexibility of the capsule shell: soft and hard capsules.

Soft capsules are single unit solid dosage forms containing a liquid or semi-solid fill. It is formed, filled and sealed in one operation by a rotary die process. For many years, soft capsules have been used as unit dose containers for liquids, while hard capsules are typically used to deliver solids in the form of powders, granules, and pills. Hard capsules are single unit dosage forms consisting of a capsule cap and a capsule body that are separately manufactured and provided in empty form for filling.

Most commonly, soft capsules are made by adding a plasticizer (usually glycerol or sorbitol) and water to gelatin. For hard capsules, the most commonly used polymer is also gelatin. The other ingredient is water which acts as a plasticizer. However, this ingredient may cause degradation of the active ingredient (e.g., bendamustine hydrochloride). Therefore, hard capsules can be additionally made from hydroxypropylmethylcellulose. The soft and hard capsules may also contain colorants and opacifiers.

A preferred type of capsule for the composition of the present invention is a hard capsule, and more particularly preferred is a gelatin hard capsule. Ideally, the material to be filled into the capsule is fluid at room temperature, which avoids heating during the filling operation. Generally, heating results in the active ingredient being susceptible to degradation.

In principle, a wide variety of excipients can be filled into hard gelatin capsules, but in addition to biopharmaceutical considerations, the chemical and physical stability of the final dosage form, as well as the dissolution behavior required to produce a safe, effective and stable dosage form, are important considerations.

In general, the formulations used to fill hard capsules may be newtonian liquids (e.g. oils, thixotropic or shear-thinning gels, or semi-solid matrix products) which are filled at elevated temperatures and in which the active ingredient is dissolved or suspended in the form of a fine dispersion. In principle, any excipient or mixture of excipients may be used, as long as the viscosity of the filling material meets the requirements of the filling process. Consistency of the capsule fill weight is important. Furthermore, the filled formulation should not show stringing and should be able to be cleanly separated from the dosing nozzle (dosingnozzle).

It has been unexpectedly found that the composition of the first embodiment of the present invention can be advantageously administered in hard gelatin capsules. If it is to be selected from the group consisting of polyethoxylated castor oil or derivatives thereof and ethylene oxide/propylene oxide block copolymers, in particular from the group consisting of glyceryl polyethylene glycol hydroxystearate, polyoxyethylene-35-castor oil andl44 or124 with bendamustine or a pharmaceutically acceptable ester, salt or solvate thereof, and filled into a gelatin hard capsule after blending, good stability, good dissolution behavior and good bioavailability are achieved. In contrast, if the glyceryl hydroxystearate is compared to a composition comprising only glyceryl polyethylene glycol hydroxystearate, for example, with bis-diglycerol polyacyladipate-1 (under the trade name Diglyceryl Polyadipate)645 commercially available) and ethylene oxide/propylene oxide block copolymers (under the trade nameL44NF or Poloxamer124 commercially available)) the dissolution behavior of bendamustine deteriorates. Furthermore, it should be noted that it is not possible to combineA25 (cetosteareth-25 (ceteareth-25) or polyethylene glycol (25) cetearyl ether) anda6 (ceteareth-6 and stearyl alcohol or polyethylene glycol (6) cetearyl ether) was used as nonionic surfactant. Other excipients commonly used in the preparation of liquid filled capsule formulations have also been shown to provide unsatisfactory results.

Furthermore, the compositions of the present invention may comprise additional excipients, in particular protective agents, such as antioxidants and antimicrobial preservatives, for example methylparaben, ethylparaben and propylparaben, as exemplified in examples 1 to 3. The antioxidant may be d-alpha-tocopherol acetate, dl-alpha-tocopherol, ascorbyl palmitate, butylated hydroxyanisole (butylated hydroxyanisole), ascorbic acid, butylated hydroxyanisole, hydroxycoumarin, butylated hydroxytoluene, ethyl gallate, propyl gallate, octyl gallate, lauryl gallate or a mixture thereof. Preferably, the antioxidant is added to a composition comprising a glyceryl polyethylene glycol hydroxystearate or polyoxyethylene-35-castor oil.

The sugar is present in the composition according to the second embodiment of the invention in a large amount, preferably in an amount of 2 to 5 times the weight of the active substance. When incorporated into the compositions of the invention, sugars have been shown to have a positive effect on the stability of the active compounds. In addition, it was unexpectedly found that these excipients lead to an increased bioavailability of the active compound, in particular bendamustine hydrochloride, compared to the reference capsules.

Preferred examples of the sugar include mannitol, maltitol, erythritol, xylitol, lactose, sucrose, glucose, sorbitol, maltose, trehalose, lactitol and dextrose (anhydrous or monohydrate).

In addition to these sugar excipients, the pharmaceutical compositions of the present invention may comprise other excipients, such as lubricants, glidants, fillers (or diluents), binders, and disintegrants, as described in more detail below.

Lubricants are substances that have one or more of the following functions during the preparation of pharmaceutical compositions, especially tablets: preventing the tablet material from adhering to the surfaces of the tablet press parts (hopper, die and punch), reducing inter-particle friction, facilitating tablet ejection from the die and improving the flow rate of the mixture (to be tabletted). The lubricant is typically selected from the group consisting of stearic acid, stearates or stearates, hydrogenated vegetable oils, magnesium oxide, polyethylene glycol, sodium lauryl sulfate and talc, and mixtures thereof. The lubricant is preferably selected from magnesium stearate, calcium stearate, zinc stearate, glyceryl palmitostearate, and sodium stearyl fumarate, and mixtures thereof. Stearic acid is the most preferred choice.

In the present application, the term glidant is understood to mean a substance which improves the flow properties of the mixture to be tabletted. For glidants, any suitable glidant may be used, such as talc, silicon dioxide and colloidal silicaStarch and calcium silicate. Silicon dioxide is generally used.

In general, the term filler (or diluent) represents those excipients which are used to increase the volume of the sheet to be compressed. This increase in size improves the handling of the solid composition. If the dosage of drug per solid composition is low, or the solid composition is too small, then a filler is generally necessary. Examples of suitable fillers are lactose, sucrose, mannitol, sorbitol, cane sugar (saccharose), starch, pregelatinized starch, microcrystalline cellulose, powdered cellulose, dibasic calcium phosphate, calcium carbonate, and any combination thereof. In a preferred embodiment, the filler is selected from the group consisting of lactose, starch, microcrystalline cellulose, microfine cellulose and any combination thereof, most preferably anhydrous lactose and microcrystalline cellulose.

Generally, the term binder is used to impart cohesiveness to the pharmaceutical formulation which ensures that the composition remains intact (especially tablets after compression molding). Different binders are used depending on the compaction technique used (direct compression, dry granulation or wet granulation). For dry compaction techniques (direct compression and dry granulation), suitable binders are lactose, sucrose, mannitol, sorbitol, cane sugar, starch, pregelatinized starch, microcrystalline cellulose, powdered cellulose, dibasic calcium phosphate, calcium carbonate and any combination thereof. In a preferred embodiment, the binder is selected from the group consisting of lactose, starch, microcrystalline cellulose, microfine cellulose and any combination thereof, most preferably anhydrous lactose and microcrystalline cellulose. In the wet granulation process, the binder may be used either as a solution or in a dry form. As suitable binders there may be mentioned, for example, polyvinylpyrrolidone, dispersible cellulose, hydroxypropyl methylcellulose, starch, pregelatinized starch, partially pregelatinized starch, gum arabic, dextrin, amylopectin and the like. Among these binders, dispersible cellulose, polyvinylpyrrolidone, hydroxypropyl cellulose and hydroxypropyl methyl cellulose are more preferable.

Disintegrants may be included in pharmaceutical compositions, particularly tablet compositions, to facilitate disintegration or disintegration of the tablet upon contact with aqueous physiological fluids. When a tablet is swallowed, a disintegrant is generally responsible for the rapid disintegration of the tablet upon contact with body fluids (e.g., saliva, gastric fluid, and intestinal fluids). Substances used as disintegrants have been classified by chemical properties into starches, celluloses, crosslinked polymers, and the like. As a result of examining the kind and the amount of the disintegrant used in carrying out the present invention, starch and modified starch (for example, sodium starch glycolate) have been found) Sodium carboxymethylcellulose and croscarmellose sodiumCrosslinked polyvinylpyrrolidone, polacrilin potassium salt (polacrilinpotassium) ((ii))IRP 88) and low-substituted hydroxypropylcellulose can produce a very good disintegrating effect.

The stability of aqueous bendamustine solutions is strongly affected by the pH value. Substantial hydrolysis of the compound is observed at pH values greater than about 5. When the pH is >5, the decomposition proceeds rapidly and within this pH range, the content of the obtained by-products is high. The major hydrolysis products are 4- [5- [ (2-chloroethyl) - (2-hydroxyethyl) amino ] -1-methyl-benzimidazol-2-yl ] -butyric acid (HP 1), 4- [5- [ bis (2-hydroxyethyl) amino ] -1-methyl-benzimidazol-2-yl ] -butyric acid (HP 2) and 4- (5-morpholinyl-1-methylbenzimidazol-2-yl) -butyric acid (HP 3):

orally administered drugs are typically absorbed in the stomach, small intestine and/or large intestine. The pH in the stomach is about 1 to 3.5, in the small intestine about 6.5 to 7.6 and in the large intestine about 7.5 to 8.0. Therefore, for compounds such as bendamustine which tend to degrade in aqueous environments at pH values above 5, it is highly preferred to have them absorbed in the stomach and not excreted to the small intestine, or even the large intestine, to avoid decomposition. There is therefore a need for pharmaceutical compositions wherein bendamustine is completely or at least to a large extent absorbed in the stomach, thereby avoiding or reducing degradation of bendamustine in the small intestine or in the large intestine.

It has been unexpectedly found that the problem can be solved by employing the pharmaceutical composition of the present invention. These compositions comprising bendamustine hydrochloride in a pharmaceutically acceptable excipient which is a non-ionic surfactant selected from the group consisting of polyethoxylated castor oil or a derivative thereof, and block copolymers of ethylene oxide and propylene oxide unexpectedly show rapid dissolution, in particular, as follows: bendamustine is at least 60% dissolved in 20 minutes, at least 70% dissolved in 40 minutes, and at least 80% dissolved in 60 minutes, and preferably at least 60% dissolved in 10 minutes, at least 70% dissolved in 20 minutes, and at least 80% dissolved in 30 minutes, as measured in artificial gastric fluid using a paddle apparatus rotating at 50rpm according to the european pharmacopoeia. The artificial gastric juice used herein means a solution prepared by dissolving 2g of sodium chloride in 1000ml of water and then adjusting the pH to 1.5. + -. 0.05 with 5N hydrochloric acid.

Furthermore, the composition was shown to be stable when subjected to accelerated stability testing. This is unexpected because the tests show that:

in the case of a reference capsule formulation comprising only bendamustine hydrochloride in a hard gelatin capsule (see reference example 1), degradation products formed within one month of storage when stored at 40 ℃/75% RH (glass vial open) and at 50 ℃. The amount of hydrolysate HP1 increased 4-fold after one month of storage with 40 ℃ and 75% RH (relative humidity) and the vial opened. For closed vials, the content of HP1 was even higher.

In the case of the capsule formulations of reference examples 2, 3 and 4, degradation products were formed within one month of storage when stored at 40 ℃/75% RH (closed glass vial), and increased with further storage.

The total time for the drug to drain from the stomach to the small intestine is between about 20 minutes and 5 hours, usually between about 30 minutes and 3 hours. Thus, the pharmaceutical compositions of the present invention advantageously reduce the degradation of bendamustine in patients, since the bendamustine therein is to a large extent released and dissolved in the stomach. Thus, the compositions of the present invention comprising bendamustine may be expected to have even improved bioavailability.

In another aspect of the invention, the oral pharmaceutical composition may be used for the treatment or prevention of relapse of a medical condition in humans or animals, preferably humans, selected from chronic lymphocytic leukemia (abbreviated CLL), acute lymphocytic leukemia (abbreviated ALL), chronic myelogenous leukemia (abbreviated CML), acute myelogenous leukemia (abbreviated AML), hodgkin's disease, non-hodgkin's lymphoma (abbreviated NHL), multiple myeloma, breast cancer, ovarian cancer, small cell lung cancer, non-small cell lung cancer, and autoimmune diseases.

In another aspect of the invention, the solid dosage form pharmaceutical composition may be used for the treatment, induction, rescue treatment, pre-transplant conditioning of stem cells, maintenance treatment, treatment of the residual disease of a human or animal, preferably a human, medical condition selected from Chronic Lymphocytic Leukemia (CLL), Acute Lymphocytic Leukemia (ALL), Chronic Myelogenous Leukemia (CML), Acute Myelogenous Leukemia (AML), hodgkin's disease, non-hodgkin's lymphoma (NHL), multiple myeloma, breast cancer, ovarian cancer, small cell lung cancer, non-small cell lung cancer, and autoimmune diseases.

The invention also comprises a method of treating a medical condition in a human or animal body selected from chronic lymphocytic leukemia, acute lymphocytic leukemia, chronic myelocytic leukemia, acute myelocytic leukemia, hodgkin's disease, non-hodgkin's lymphoma, multiple myeloma, breast cancer, ovarian cancer, small cell lung cancer, non-small cell lung cancer and autoimmune diseases, or preventing the recurrence of said medical condition, comprising administering an effective amount of a pharmaceutical formulation of the invention to a human or animal body in need of treatment. A preferred medical condition is non-hodgkin's lymphoma.

In another aspect of the invention, the pharmaceutical composition may be administered in combination with at least one additional active agent, wherein the additional active agent is administered prior to, simultaneously with, or after administration of the pharmaceutical composition. The at least one other active agent is preferably a CD 20-specific antibody (e.g. rituximab or ofatumumab), an anthracycline derivative (e.g. doxorubicin or daunorubicin), a vinca alkaloid (e.g. vincristine), a platinum derivative (e.g. cisplatin or carboplatin), daphnolide (FK 866), YM155, thalidomide and its analogues (e.g. lenalidomide), or a proteasome inhibitor (e.g. bortezomib).

The pharmaceutical compositions of the present invention may also be administered in combination with at least one corticosteroid, wherein the corticosteroid is administered prior to, concurrently with, or subsequent to the administration of the pharmaceutical composition. Examples of such corticosteroids are prednisone, prednisolone and dexamethasone.

Some dosing regimens are possible. For example, in patients with NIn patients with HL, the following doses can be administered: bendamustine administered in an oral formulation at a total amount of 200 mg/person/day on days 1-5 + vincristine administered intravenously at 2mg on day 1 + vincristine administered intravenously at 100mg/m on days 1-5²Prednisone, once every 3 weeks. In patients with MM, administration may be as follows: bendamustine in a total amount of 400-500 mg/person/day administered in an oral formulation on days 1 and 2 + 60mg/m administered intravenously or orally on days 1-4²Prednisone, once every 4 weeks. In patients with CCL, administration can be as follows: on days 1 and 2, bendamustine is administered in an oral formulation in a total amount of 200-300 mg/person/day once every 4 weeks + on days 1-4, either intravenously or orally at 60mg/m²Once every 4 weeks.

The liquid-filled hard gelatin capsules of the present invention are also advantageous in that there is no need to provide a coating for the active ingredient (which may optionally be mixed with one or more excipients) to further mask the taste of the active ingredient and/or to protect the active ingredient from possible deleterious effects (e.g., oxidation, degradation) caused by light and/or moisture, or to protect the oral mucosa of the subject from damage due to interaction with the active ingredient.

The invention is further illustrated by the following examples. It will be apparent to those skilled in the art that these examples are for illustrative purposes only and should not be construed as limiting the invention.

Examples of the present invention

A) Examples relating to the first embodiment of the present invention

1. Capsule preparation

Reference example 1: bendamustine capsule formulations (prior art)

20.0. + -. 1mg of bendamustine hydrochloride was weighed into the capsule body of an empty gelatin hard capsule, and then placed into an Agilent HPLC vial (6 ml) made of transparent glass. The capsule is closed by placing the capsule cap over the capsule body and gently pressing.

The capsules were stored at 40 ℃/75% RH (glass vial open) or 50 ℃ (glass vial closed). The content of bendamustine hydrochloride and related substances was measured by HPLC (column: ZorbaxBonus-RP,5 μm; column oven temperature: 30 ℃; autosampler temperature: 5 ℃; detector: 254 nm). The results are shown in table 1:

^*1: NP 1: 4- [6- (2-chloroethyl) -3,6,7, 8-tetrahydro-3-methyl-imidazo [4,5-h]-[1,4]Benzothiazin-2-yl radical]Butyric acid

BM1 dimer: 4- {5- [ N- (2-chloroethyl) -N- (2- {4- [ 5-bis (2-chloroethyl) amino-1-methylbenzimidazol-2 yl ] butanoyloxy } ethyl) amino ] -1-methylbenzimidazol-2 yl } butanoic acid

BM1EE:4- [5- [ bis (2-chloroethyl) amino ] -1-methyl-benzimidazol-2-yl ] butanoic acid ethyl ester

^*2: n.d.: non-detectable, i.e. exceeding the detection limit (area percentage less than 0.05%)

Reference example 2

For a 1000 capsule batch, all excipients except colloidal silica and stearic acid were filled into a Somakon container (5L). Bendamustine was added and mixing was performed at 1000rpm for 4 minutes (wiper 10 rpm). The resulting mixture was sieved through a 0.5mm screen. The mixture was refilled into the container and colloidal silica was added. Mixing was carried out under the above conditions for 2 minutes. Thereafter stearic acid was added and mixing was continued for 1 minute. The mixture was then sieved through a 0.5mm screen, reloaded into the container and mixed for an additional 30 seconds, all under the same conditions as described above.

The mixture was transferred to a capsule filler (ZanassiAZ 5) and filled into hard gelatin capsules (size 2) (average mass: 259.5mg (initial) -255.3mg (final)) and hypromellose capsules (size 2) (average mass: 255.8mg (initial) -253.4mg (final)), respectively. The capsules were stored in closed glass vials at 40 ℃/75% RH. The amounts of bendamustine hydrochloride and related substances such as degradation products, by-products of synthesis were measured by HPLC (column: ZorbaxBonus-RP,5 μm; column oven temperature: 30 ℃; autosampler temperature: 5 ℃; detector: 254 nm). The results are shown in table 2b (filled in hypromellose capsules) and table 2c (filled in gelatin capsules).

^*3: unidentified compound peak at relative retention time of 0.65 relative to the main peak

Reference example 3

For 1000 capsules, all excipients except colloidal silica and stearic acid were filled into a Somakon container (5L). Bendamustine was added and mixing was performed at 1000rpm for 4 minutes (wiper 10 rpm). The resulting mixture was sieved through a 0.5mm screen. The mixture was refilled into the container and colloidal silica was added. Mixing was carried out under the above conditions for 2 minutes. Thereafter stearic acid was added and mixing was continued for 1 minute. The mixture was then sieved through a 0.5mm screen, reloaded into the container and mixed for an additional 30 seconds, all under the same conditions as described above.

The mixture was transferred to a capsule filler (ZanassiAZ 5) and filled into hard gelatin capsules (size 2) (average mass: 257.9mg (initial) -255.2mg (final)) and hypromellose capsules (size 2) (average mass: 261.1mg (initial) -257.8mg (final)), respectively. The capsules were stored in closed glass vials at 40 ℃/75% RH. The amount of bendamustine hydrochloride and related substances was measured by HPLC as described above. The results are shown in table 3b (filled into hypromellose capsules) and table 3c (filled into gelatin capsules).

Reference example 4

For 1000 capsules, all excipients except colloidal silicon dioxide and magnesium stearate were filled into Somakon containers (2.5L). Bendamustine was added and mixing was performed at 1000rpm for 4 minutes (wiper 10 rpm). The resulting mixture was sieved through a 0.5mm screen. The mixture was refilled into the container and colloidal silica was added. Mixing was carried out under the above conditions for 2 minutes. Magnesium stearate was then added and mixing continued for 1 minute. The mixture was then sieved through a 0.5mm screen, reloaded into the container and mixed for an additional 30 seconds, all under the same conditions as described above.

The mixture was transferred to a capsule filler (ZanassiAZ 5) and filled into hard gelatin capsules (size 2) (average mass: 241.3mg (start) -244 mg (end)) and hypromellose capsules (size 2) (average mass: 243.5mg (start) -243 mg (end)), respectively. The capsules were stored in closed glass vials at 40 ℃/75% RH. The content of bendamustine hydrochloride and related substances was measured by HPLC as described above. The results are shown in table 4b (filled into hypromellose capsules) and table 4c (filled into gelatin capsules).

Example 1

0.68g of methylparaben, 0.068g of propylparaben and 0.068g of butylhydroxymethylbenzol were weighed out and dissolved in 6.14g of ethanol. In a sufficient amountRH40 melted at 40 ℃. 5.56g of the resulting ethanol solution, 36.83g of moltenRH40 and 202.82gL44NF and mixed with a mechanical stirrer at 800rpm until the mixture became clear. The mixture was allowed to solidify by placing it at 10 ℃. 24.80g bendamustine hydrochloride was then added to the coagulated mixture by manual stirring, followed by homogenization with an UltraturraxT18 high speed homogenizer at 15500rpm for 10 minutes to distribute the bendamustine hydrochloride uniformly throughout the mixture. The homogenized suspension was filled into hard gelatin capsules using a CFS1200 capsule filling and sealing machine operating at 25 ℃. The capsule is closed and sealed. The resulting liquid-filled capsules were stored at 40 ℃/75% RH, 30 ℃/65% RH, 25 ℃/60% RH, and 5 ℃ in closed amber glass vials with screw stoppers. The amounts of bendamustine hydrochloride and related substances such as degradation products, by-products of synthesis were measured by HPLC (column: ZorbaxBonus-RP,5 μm; column oven temperature: 30 ℃; autosampler temperature: 5 ℃; detector: 254 nm). The results are shown in table 5 b.

Example 2

0.68g of methylparaben, 0.068g of propylparaben and 0.068g of butylhydroxymethylbenzol were weighed out and dissolved in 6.14g of ethanol. In a sufficient amountRH40 melted at 40 ℃. 5.56g of the resulting ethanol solution and 239.65g of moltenRH40 and mixed with a mechanical stirrer at 800rpm until the mixture becomes transparent. The mixture was allowed to solidify and cool to room temperature. 24.80g bendamustine hydrochloride was then added to the coagulated mixture by manual stirring, followed by homogenization with an UltraturraxT18 high speed homogenizer at 15500rpm for 10 minutes to distribute the bendamustine hydrochloride uniformly throughout the mixture. The homogenized suspension was filled into hard gelatin capsules using a CFS1200 capsule filling and sealing machine operating at 40 ℃. The capsule is closed and sealed. The liquid-filled capsules obtained above were stored at 40 ℃/75% RH, 30 ℃/65% RH, 25 ℃/60% RH and at 5 ℃ in closed amber glass bottles with screw stoppers. As described above, the amount of bendamustine hydrochloride and related substances such as degradation products, by-products of synthesis, and the like were measured by HPLC. The results are shown in table 6b.

Example 3

0.68g of methylparaben, 0.068g of propylparaben and 0.068g of butylhydroxymethylbenzol were weighed out and dissolved in 6.14g of ethanol. In a sufficient amountRH40 melted at 40 ℃. 5.56g of the resulting ethanol solution, 36.83g of moltenRH40 and 202.82g645, and mixed with a mechanical stirrer at 800rpm until the mixture becomes transparent. The mixture was allowed to solidify by placing it at 10 ℃. 24.80g bendamustine hydrochloride was then added to the coagulated mixture by manual stirring, followed by homogenization with an UltraturraxT18 high speed homogenizer at 15500rpm for 10 minutes to distribute the bendamustine hydrochloride uniformly throughout the mixture. The homogenized suspension was filled into hard gelatin capsules using a CFS1200 capsule filling and sealing machine operating at 30 ℃. The capsule is closed and sealed. The resulting liquid-filled capsules were stored at 40 ℃/75% RH, 30 ℃/65% RH, 25 ℃/60% RH, and 5 ℃ in closed amber glass vials with screw stoppers. As described above, the amount of bendamustine hydrochloride and related substances such as degradation products, by-products of synthesis, and the like were measured by HPLC. The results are shown in table 7b.

Example 4

2. Disintegration and dissolution test

Example 5

The liquid-filled capsule formulations of examples 1, 2 and 3 were subjected to disintegration testing in 1000.0ml of buffer, pH =1.0 ± 0.05, using a disintegration apparatus a operating at 37.0 ℃ ± 0.5 ℃. The results are listed in tables 8a, 8b and 8 c.

Example 6

The liquid filled capsule formulations of examples 1, 2 and 3 were subjected to a dissolution test in an artificial gastric acid solution at a pH of 1.5 (see european pharmacopoeia PhEur: 2.9.3: dissolution test of solid dosage forms in recommended dissolution media).

The dissolution samples were tested for analysis by HPLC (column: ZorbaxBonus-RP,5 μm; column oven temperature: 30 ℃; autosampler temperature: 5 ℃; detector: 254 nm). Artificial gastric juice (pH 1.5) was prepared by the following method: 250.0ml of 0.2M potassium chloride (0.2M) was added to a 1000ml volumetric flask, 207.0ml of 0.2M hydrochloric acid was added, followed by dilution to 1000ml with Milli-Q water. If necessary, the pH is measured and adjusted to 1.5. + -. 0.05 with 2N hydrochloric acid or 2N potassium hydroxide.

Dissolution tests were carried out according to section 2.9.3 of the european pharmacopoeia 6.0 using apparatus 2 (paddle apparatus). The paddle speed was 50rpm, the temperature was 37 ℃. + -. 0.5 ℃ and the amount of dissolution medium was 500 ml.

The test results for the liquid-filled hard capsules of examples 1, 2 and 3 are shown in tables 9a, 9b and 9 c:

as can be seen from the above tables 9a, 9b and 9c, only the liquid filled hard capsule formulation of example 2 of the present invention showed the following preferred rapid dissolution behavior of bendamustine, namely: bendamustine dissolves at least 60% in 10 minutes, at least 70% in 20 minutes and at least 80% in 30 minutes as measured according to the european pharmacopoeia in 500ml artificial gastric fluid using a paddle apparatus rotating at 50 rpm.

Example 7

Table 10: analytical test results for the formulation of example 4

3. In vivo assay

Example 8

The liquid-filled hard capsules of example 2 containing 50mg bendamustine were orally administered to male and female beagle dogs and compared to the capsules of reference example 1 to determine the bioavailability (AUC and Cmax) of the 1 dose (i.e., 50 mg) of bendamustine and the fluctuation levels of the bioavailability (i.e.,% CV of AUC and Cmax) of these capsule formulations. The test also included other formulations (formulation X), but since this formulation was not within the scope of the present invention, a detailed description is not provided. The total number of animals required was 16. The basic trial was designed as a cross-over design of 8 animals per arm (arm).

Phase 1 (Single dose capsule, day one)

There was a one week washout period (wash-out).

Phase 2 (one week after phase 1, single dose of each of the following formulations, day 8)

The mean plasma distribution versus time for the capsule formulation (reference 1) and the liquid-filled capsule formulation of example 2 is shown in figure 1.

Example 9

An open-label, randomized, two-way crossover study to evaluate the absolute bioavailability of oral bendamustine in cancer patients was performed to evaluate the absolute bioavailability of bendamustine (example 2) administered in an oral formulation. In addition to evaluating the pharmacokinetics of bendamustine in plasma after oral and intravenous administration, the objective was to evaluate the safety and tolerability of bendamustine after intravenous administration, especially oral administration of the formulation of example 2.

6 patients were hospitalized for two periods of treatment: day 1 to day 2 (stage 1) and day 7 to day 9 (stage 2). Patients were housed to receive one of the following two treatments in randomized order on days 1 and 8:

-a single oral dose of 110.2mg (2 x55.1 mg) bendamustine hydrochloride (HCl) equivalent to about 100mg bendamustine free base, and

a single intravenous dose of 100mg bendamustine hydrochloride, which is equivalent to 90.7mg bendamustine free base.

The dose of bendamustine hydrochloride was selected based on the safety of the oral formulation in preclinical studies and the safety of the registered intravenous formulations (intravenous dose of 100mg, oral dose of 110.2 mg).

Blood samples were collected on days 1, 2, 8 and 9 to determine the pharmacokinetics of bendamustine and its metabolites in plasma after oral and intravenous administration. The time points after intravenous bendamustine administration were selected according to data in the literature (Preiss 1985). Preiss and colleagues reported that the mean bioavailability of bendamustine after oral administration of bendamustine in capsule form at doses ranging from 250mg to 350mg to cancer patients was 57% (range: 25% to 94%;% CV = 44%). Single doses of bendamustine (100 mg intravenously or 110.2mg orally with bendamustine hydrochloride) were administered orally or intravenously in the morning of day 1 and morning of day 8.

Bendamustine is administered orally as two liquid filled hard shell capsules with 250ml of water, or as an intravenous infusion for 30 minutes.

Prior to oral or intravenous bendamustine administration in the morning, patients were required to fasted overnight for at least 8 hours, except that water was available (up to two hours prior to administration of the study drug). After 2 hours of each administration, the patient may eat a small breakfast.

Total dosing period was 6 days (day 1 to day 2 and day 7 to day 9) excluding screening and follow-up after the study.

Patients were prohibited from taking certain medications within two weeks prior to the first study medication administration.

The blood concentration-time curves obtained after evaluation of 6 patients are shown in fig. 2. By formula AUC_{Through the mouth}dose/AUC_{Through the vein}The absolute bioavailability calculated per dose x 100% was 58.5% on average, with a standard deviation of 9.3, and an inter-individual difference (expressed as% CV) of 15.9.

Thus, the bioavailability of bendamustine hydrochloride in the oral formulation of example 2 was observed to be consistent with the bendamustine-containing capsules reported in the previous literature (Preiss), but the inter-patient variability was greatly reduced.

B) Examples relating to the second embodiment of the present invention

1. Compatibility test

Example 10a

A compatibility test mixture was prepared containing 1:1 (mass/mass) bendamustine hydrochloride and excipients. The excipient is selected from mannitol and lactose. After preparation, the mixture was charged into Agilent (Agilent) HPLC vials (6 ml) made of clear glass and stored under different storage conditions as shown in table 11 below. At defined time points, samples were taken from the storage conditions and checked for purity (HPLC, column: ZorbaxBonus-RP,5 μm; column oven temperature: 30 ℃; autosampler temperature: 5 ℃; detector: 254 nm) and appearance.

Table 11: storage conditions

^*Storing at 50 deg.C for one month before storing at 70 deg.C

^**Storage at 25 ℃/60% r.h. for one month before storage at 40 ℃/75%

In all these mixtures, the bendamustine hydrochloride content (measured by HPLC) remained almost unchanged and always above 99% for all three storage conditions. For all three storage conditions, almost no hydrolysate HP1 (area% < 0.2) was detected.

Visually inspecting the bendamustine hydrochloride mixture. All tested mixtures were in specification and appeared as white to off-white powders when tested immediately after preparation and after one month storage under all three storage conditions.

Example 10b

To further perform the compatibility test according to the method of example 1a, a mixture was prepared comprising 1:1 (mass/mass) bendamustine hydrochloride and excipients. The excipient is selected fromEPO, sodium carboxymethylcelluloseRC 591) and Crospovidone (Crospovidone).

In useIn the case of EPO, the initial amounts of the impurities HP1 (hydrolysate) and BM1DIMER were greatly increased (HP 1:1.5%, BM1 DIMER: 1%), but under all storage conditions excluding the effect of humidity, a reduction in these impurities during storage could be detected. In the case of cross-linked polyvinylpyrrolidone, a significant increase from 0.1% to 0.4% of HP1 could be detected under storage conditions of 40 ℃/75% r.h./vial open. Under all other storage conditions (vial closure), no increase in HP1 was detected.

Under 70 ℃/vial closed storage conditions, comprisingThe appearance of the mixture of EPO and the mixture comprising crosslinked polyvinylpyrrolidone changed. Both mixtures became slightly viscous. Furthermore, the color of the mixture comprising crosslinked polyvinylpyrrolidone changed from white to cream.

Under 70 ℃/vial closed storage conditions, comprisingAnd a mixture comprisingThe color of the mixture of RC591 also changes to cream.

2. Tablet formulation

Example 11

253g of a mixture containing mannitol (as main excipient) and microcrystalline cellulose in the relative amounts shown in Table 2a below, were prepared by mixing for 15 minutes in a1 liter cube blender (Erweka),Colloidal silica, talc and stearic acid. 10.612g of the mixture and 3.0g of bendamustine hydrochloride were then sieved through a 0.425mm sieve, then transferred to a Turbula mixer T2A equipped with a 50ml glass vial, and then mixed at 60rpm for 10 minutes.

The resulting mixture was compression molded into round tablets having the following characteristics:

average diameter: 9.1 mm; average mass: 247.7 mg; average hardness: and 81N.

The tablets were stored at 40 ℃/75% RH (glass vial open) or at 50 ℃ (glass vial closed). The amounts of bendamustine hydrochloride and related substances such as degradation products, by-products of synthesis were measured using HPLC (column: ZorbaxBonus-RP,5 μm; column oven temperature: 30 ℃; autosampler temperature: 5 ℃; detector: 254 nm). The results are shown in table 12 b.

^*1: NP 1: 4- [6- (2-chloroethyl) -3,6,7, 8-tetrahydro-3-methyl-imidazo [4,5-h]-[1,4]Benzothiazin-2-yl radical]Butyric acid

BM1dimer, 4- {5- [ N- (2-chloroethyl) -N- (2- {4- [ 5-bis (2-chloroethyl) amino-1-methylbenzimidazol-2-yl ] butanoyloxy } ethyl) amino ] -1-methylbenzimidazol-2-yl } butanoic acid

BM1EE Ethyl 4- [5- [ bis (2-chloroethyl) amino ] -1-methyl-benzimidazol-2-yl ] butanoate

^*2: n.d.: non-detectable, i.e. exceeding the detection limit (area percentage less than 0.05%)

Example 12

Mixtures and tablets were prepared in the same manner as described in example 11, using the compounds and relative amounts described in table 3a below.

The tablet has the following characteristics:

average diameter: 9.1 mm; average mass: 248.9 mg.

The tablets were stored at 40 ℃/75% RH (glass vial open) or at 50 ℃ (glass vial closed). The content of bendamustine hydrochloride and related substances was measured by HPLC as described above. The results are shown in table 3 b:

example 13

Tablets were prepared in the same manner as described in example 11, using the compounds and relative amounts described in table 14a below.

The tablet has the following characteristics:

average diameter: 9.1 mm; average mass: 247.8 mg.

The tablets were stored at 40 ℃/75% RH (glass vial open) or at 50 ℃ (glass vial closed). The amount of bendamustine hydrochloride and related substances was measured by HPLC as described above. The results are shown in table 14 b:

reference example of the prior art

20.0. + -. 1mg of bendamustine hydrochloride was weighed into the capsule body of an empty gelatin hard capsule, and then placed into an Agilent HPLC vial (6 ml) made of transparent glass. The capsule is closed by placing the capsule cap over the capsule body and gently pressing. The capsules were stored at 40 ℃/75% RH (glass vial open) or 50 ℃ (glass vial closed). The amount of bendamustine hydrochloride and related substances was measured by HPLC as described above. The results are shown in table 15:

it is evident that, although the capsule formulation was prepared from pure bendamustine hydrochloride without any further processing steps, the stability of the capsule formulation was much poorer compared to the tablet formulation of the present invention. More degradation products were formed both at 40 ℃/75% RH (glass vial open) and within one month of storage at 50 ℃ (glass vial closed). After one month of storage at 40 ℃, 75% RH (relative humidity) with the vial open, the amount of hydrolysate HP1 increased 4-fold. For the closed vials, the HP1 content was even higher, perhaps due to reaction with the capsules. In summary, tablets provide a much more stable solid dosage form compared to capsules.

Example 14

8.0g of hydroxypropylmethylcellulose and 1.5g of PEG6000 were dissolved in 88.5g of purified water. Thereafter, 2.0g of yellow iron sesquioxide and 0.5g of titanium oxide were dispersed therein to produce a coating liquid. The tablets obtained in example 11 were coated using a film coater in an amount of 3% of the solution per mass of the tablet.

Example 15

Preparation method of 1000 tablets

All core ingredients except colloidal silica and stearic acid were loaded into a Somakon vessel (5L). Bendamustine was added and mixing was performed at 1000rpm for 4 minutes (wiper 10 rpm). The resulting mixture was sieved through a 0.5mm screen. The mixture was refilled into the container and colloidal silica was added. Mixing was carried out under the above conditions for 2 minutes. Thereafter stearic acid was added and mixing was continued for 1 minute. The mixture was then sieved through a 0.5mm screen, reloaded into the container and mixed for an additional 30 seconds, all under the same conditions as described above.

Compression molding the mixture into round tablets having the following characteristics:

average diameter: 9.5 mm; average mass: 254.6mg (starting) to 257.2mg (final); friability 0.1%; average hardness: 122N (start) to 128N (final).

Thereafter usingDispersion the tablets were film coated until the mass increased by 5%.

The average mass of the film-coated tablets was 268.4 mg.

The core and film coated tablets were stored in closed amber glass vials at 40 ℃/75% RH. As described above, the amount of bendamustine hydrochloride and related substances such as degradation products, by-products of synthesis, and the like were measured by HPLC. The results are shown in tables 16b.1 and 16 b.2.

^*3: unidentified compound peak at relative retention time of 0.69 relative to the main peak

Example 16

Preparation method of 1000 tablets

All core ingredients except colloidal silica and stearic acid were loaded into a Somakon vessel (5L). Bendamustine was added and mixing was performed at 1000rpm for 4 minutes (wiper 10 rpm). The resulting mixture was sieved through a 0.5mm screen. The mixture was refilled into the container and colloidal silica was added. Mix for 2 minutes under the above conditions. Thereafter stearic acid was added and mixing was continued for 1 minute. The mixture was then sieved through a 0.5mm screen, reloaded into the container and mixed for an additional 30 seconds, all under the same conditions as described above.

Compression molding the mixture into round tablets having the following characteristics:

average diameter: 9.5 mm; average mass: 262.4mg (initial) to 254.4mg (final); friability: 0.1% (initial) to 0.2% (final); average hardness: 98N (initial) to 91N (final).

Thereafter usingDispersion the tablets were film coated until the mass increased by 3%.

The average mass of the film-coated tablets was 273.5 mg.

The core and film coated tablets were stored in closed amber glass vials at 40 ℃/75% RH. The content of bendamustine hydrochloride and related substances was measured by HPLC as described above. The results are shown in tables 17b.1 and 17 b.2:

example 17

Preparation method of 1000 tablets

All core ingredients except colloidal silica and stearic acid were loaded into a Somakon vessel (2.5L). Bendamustine was added and mixing was performed at 1000rpm for 4 minutes (wiper 10 rpm). The resulting mixture was sieved through a 0.5mm screen. The mixture was refilled into the container and colloidal silica was added. Mixing was carried out under the above conditions for 2 minutes. Thereafter stearic acid was added and mixing was continued for 1 minute. The mixture was then sieved through a 0.5mm screen, reloaded into the container and mixed for an additional 30 seconds, all under the same conditions as described above.

Compression molding the mixture into round tablets having the following characteristics:

average diameter: 9.5 mm; average mass: 252.2mg (start) to 250.7mg (end); friability: 0.1% (initial) to 0.2% (final); average hardness: 65N (initial) to 73N (final).

Thereafter usingDispersion the tablets were film coated until the mass increased by 3%.

The average mass of the film-coated tablets was 253.6 mg.

The core and film coated tablets were stored in closed amber glass vials at 40 ℃/75% RH. The amount of bendamustine hydrochloride and related substances was measured by HPLC as described above. The results are shown in tables 18b.1 and 18 b.2:

example 18

The preparation method of the 600 tablet preparation PF1 comprises the following steps:

33.06g bendamustine, 111.60g dextrose, 40.92g lactose, 11.22g microcrystalline cellulose, and 1.20g magnesium stearate were weighed and transferred to a double-layer polyethylene bag and mixed for 5 minutes. The powder mixture was then transferred to the hopper of an eccentric tablet press (KorschEK 0) and compression molded into round tablets with the following characteristics: average diameter: 10.0 mm; average mass: 336.9mg (initial) to 335.98 (final); friability: 0.15 percent; average hardness value: 69.25N (initial) to 68.60N (final).

Then in a coating pan (4M 8 ForMatePara coat), 9% white Opadry (A) (B)TMWhite) aqueous suspension the core was coated and dried. The average mass of the tablets was 342.42 mg. The tablets are then filled into amber glass bottles closed with screw stoppers andstored at 40 ℃/75% RH.

The preparation method of the 600 tablet preparation PF2 comprises the following steps:

33.06g bendamustine, 111.42g lactose, 39.60g trehalose, 12.60g crospovidone, and 1.32g magnesium stearate were weighed and transferred to a double-layer polyethylene bag and mixed for 5 minutes. The powder mixture was then transferred to the hopper of an eccentric tablet press (KorschEK 0) and compression molded into round tablets with the following characteristics: average diameter: 10.0 mm; average mass: 332.95mg (starting) to 332.12 (final); friability: 0.3 percent; average hardness value: 65.9N (initial) to 59.0N (final).

Then in a coating pan (4M 8 ForMateParat), use 9%Aqueous TMWhite suspension cores were coated and dried. The average mass of the tablets was 340.1 mg. The tablets were then filled into amber glass vials closed with screw stoppers and stored at 40 ℃/75% RH.

Preparation method of the preparation PF 3:

sorbitol and anhydrous dextrose were weighed. 140.64g of sorbitol were dissolved in 105.48g of purified water, and 659.36g of dextrose were subsequently granulated in a fluid bed granulator (4M 8 ForMateFluidBed) using the resulting solution. Thereafter, the granules were dried at 60 ℃ and sieved through a 850 μm sieve.

33.06g of bendamustine hydrochloride, 149.82g of sorbitol/dextrose granules, 13.8g of microcrystalline cellulose and 1.32g of magnesium stearate were weighed out and transferred to a double-layer polyethylene bag to be mixed for 5 minutes. The powder mixture was then transferred to the hopper of an eccentric tabletting machine (KorschEK 0) and compression molded into round tablets with an average diameter of 10.0 mm. The average mass of the tablets was 335.99mg (starting) to 339.50 (final); the friability is 0%; the average hardness values ranged from 125.60N (initial) to 129.7N (final). The tablets were then subjected to a two step pre-treatment process (only on selected batches): the tablets were placed at 25 ℃/60% r.h. for two hours, followed by two hours at 40 ℃.

Then in a coating pan (4M 8 ForMateParat), use 9%The aqueous TMWhite suspension coats the tablets. The average mass of the tablets was 341.43 mg. The tablets were then filled into amber glass vials closed with screw stoppers and stored at 40 ℃/75% RH.

The amount of bendamustine hydrochloride and related substances in the preserved film-coated tablets was measured by HPLC as described above. The results are shown in tables 19b.1 to 19 b.3:

3. dissolution test

Example 19

The tablet formulations of examples 11 and 12 were subjected to dissolution test in artificial gastric fluid at T = 0. The dissolution samples were tested for analysis by HPLC (column: ZorbaxBonus-RP,5 μm; column oven temperature: 30 ℃; autosampler temperature: 5 ℃; detector: 254 nm). Artificial gastric juice (pH 1.5) was prepared by the following steps: 2g of sodium chloride p.A. are dissolved in 1000ml of water and the pH is adjusted to 1.5. + -. 0.05 with 5N hydrochloric acid. Dissolution tests were carried out according to the european pharmacopoeia 6.0, chapter 2.9.3, using apparatus 2 (paddle apparatus). The paddle speed was 50rpm, the temperature was 37 ℃. + -. 0.5 ℃ and the amount of dissolution medium was 500 ml.

The tablet formulation results for example 11 (tablet formulation 1) and example 12 (tablet formulation 2) are shown in table 20a below:

table 20 a:

the results of the same dissolution tests performed on the coated tablet formulations of example 15, example 16 and example 17 at T =0 are shown in table 20b below:

TABLE 20b

The corresponding dissolution data for the tablets of example 18 were:

as can be seen from the above, all tablet formulations of the present invention show rapid dissolution behavior of bendamustine. In particular, the formulations of the present invention exhibit bendamustine dissolution behavior as defined herein before.

4. In vivo assay

Animal bioavailability studies of bendamustine in beagle dogs: outline of PK study

Study experiment 1

The study objective was to determine the bioavailability of bendamustine at 1 dose (i.e., 50 mg) in 3 tablet formulations (T1-3) and 1 capsule formulation (C) (4 total oral formulations): AUC and Cmax

Total number of animals required: 16 pieces of

Basic design:

cross-over design, 8 animals per branch (arm):

table 21 a: phase 1 (single dose tablet, or capsule, day 1):

elute one week (wash-out)

Table 21 b: phase 2 (one week after phase 1, single dose of each of the following formulations, day 8):

elute in one week

Table 21 c: phase 3 (one week after phase 2, single dose of each of the following formulations, day 15):

study experiment 2

The objective of the study was to determine the bioavailability of bendamustine at 1 dose (i.e., 50 mg) in 1 tablet formulation T4 and 1 capsule formulation (C) (3 total oral formulations): AUC and Cmax

Total number of animals required: 16 pieces of

Basic design:

crossover design, 8 animals per branch:

table 22 a: stage 1 (Single dose capsule, day 1)

Elute in one week

Table 22 b: phase 2 (one week after phase 1, single dose of each of the following formulations, day 8):

example 20

The coated tablet of example 18 (formulation 3, with 50mg bendamustine) containing was orally administered to male and female dogsCoating, tablet T4) and compared to the capsules of the reference example.

The mean plasma concentrations of the capsule formulation and the coated tablet of example 18 are shown over time in figure 3.

Example 21

The coated tablets of examples 15, 16 or 17 (tablets T1 to T3) containing 50mg bendamustine were orally administered to male and female dogs, and compared to the capsules of the reference example.

The mean plasma-time relationship of the capsule formulations and the coated tablets of examples 15-17 are shown in figure 3.

The experiments were performed for the following:

-assessing which sugar or sugar mixture is suitable for obtaining a chemically stable tablet with fast dissolution behaviour and a hardness value suitable for coating;

-evaluating the compatibility between API and excipient;

-developing placebo and API containing batches by studying the following different preparation processes: dry granulation, direct pressing and wet granulation;

-evaluating different bendamustine hydrochloride/sugar weight ratios;

-assessing the effect of sugar purity on bendamustine hydrochloride impurity formation;

study of the effect of moisture content on the technical performance and stability of the tablets prepared;

use of commercially available lyophilized bendamustine hydrochloride productTablets were prepared and the properties of these tablets were compared to those of tablets prepared using a comparable amount of mannitol and bendamustine hydrochloride.

Tablets of the invention containing 50mg bendamustine (55 mg for bendamustine hydrochloride) were prepared using the following sugars.

TABLE 23

The quality of the prepared batches was assessed by observing physical appearance, identification tests (HPLC), dissolution tests, content and related substance analysis (HPLC), content uniformity tests (HPLC), hardness tests and water content (karl fischer method). The batches were filled into amber glass bottles and accelerated stability studies were performed under storage conditions as detailed in the table below. For each batch prepared containing API, some tablets were stored at 5 ℃ as backup samples.

Next, various excipients associated with the tablet manufacturing process were investigated. By using these excipients, several placebo preparation trials were carried out using dry granulation, thus obtaining preliminary information about the preparation methods suitable for obtaining tablets with good quality.

Two types of disintegrants are used: microcrystalline cellulose as a standard disintegrant: (PH 112), and crosslinked polyvinylpyrrolidone used only for lot D001T/002Selection for batch D001T/002 (Filler: Anhydrous lactose)Based on the similarity between this formulation and the prototype formulation of example 9. For all batches prepared, magnesium stearate was used as a lubricant. The dry granulation preparation process for placebo testing included the following steps:

1. accurately weighing sugar and a partial amount of lubricant (of the total amount)83.3%_w/w) And then mixed in a polyethylene bag for 2 minutes.

2. The resulting mixture was compacted using a tablet press equipped with 18mm diameter punches.

3. The resulting billet (slug) was sieved using a 850 micron screen.

4. The granules were weighed and mixed with disintegrant and remaining amount of lubricant (16.7%_w/w) Mixing was carried out for 2 minutes, after which tabletting was carried out with a punch of 10mm diameter.

Tables 24 and 25 summarize the composition of each placebo formulation and the results of the analytical tests performed on the final mixture and tablets. The results observed during the preparation of the placebo batches and/or during their analytical characterisation are shown in table 16.

The results of the analyses and physical tests carried out on placebo batches D001T/001, D001T/002, D001T/004, D001T/013, D001T/015 show that these formulations are suitable for preparation by dry granulation and for further studies by addition of API. All other formulations are characterized by a difficulty in compacting the powder and by a high friability of the tablets in the case of tablets obtained.

Lot D001T/005 (Filler: β -Cyclodextrin) showed good behavior during dry preparation, high hardness, low friability, but long disintegration timeAnd an API was added to further study this formulation (see below).

The prepared batches are granulated by a dry method according to the weight ratio of the bendamustine hydrochloride to the sugar of 1:5

A placebo formulation rated more suitable for preparing tablets containing an Active Pharmaceutical Ingredient (API) by dry granulation was modified to contain an API, and two API/sugar weight ratios were explored: 1:5 and 1: 2.

In this paragraph, a formulation containing API/sugar in a weight ratio of 1:5 is described.

Two types of disintegrants are used: microcrystalline cellulose as a standard disintegrant: (PH 112), and crosslinked polyvinylpyrrolidone used only for lot D001T/022For all batches prepared, magnesium stearate was used as a lubricant.

The process for preparing the API-containing batch by dry granulation comprises the following steps:

1. sugar, a portion of the lubricant (83.3% of the total amount) was weighed accurately_w/w) And bendamustine hydrochloride, and then mixed in a double-layer polyethylene bag for 5 minutes.

2. The powder mixture was compressed using a tablet press equipped with 18mm diameter punches.

3. The resulting billet was sieved using a 850 micron sieve to obtain granules.

4. The granules were weighed and mixed with disintegrant and remaining amount of lubricant (16.7%_w/w) Mix in a double polyethylene bag for 5 minutes.

5. The resulting mixture was tableted using a 10mm diameter punch.

Table 27 summarizes the composition of each of the API-containing formulations prepared, and the results of the analytical tests performed on the final mixtures containing the API; table 28 summarizes the results of the analytical tests performed on the obtained articles.

TABLE 27 Dry granulation-API/sugar weight ratio 1: 5. Composition and analytical results of the final mixture of API-containing batches.

TABLE 28 Dry granulation-API/sugar weight ratio 1: 5. Analysis results of the tablets of the API-containing batches.

The results of analytical tests carried out on the final mixture and on the resulting preparation (mainly content uniformity and purity) are in most cases good. All API-containing batches showed satisfactory quality uniformity, API content uniformity and low impurity content. The impurity profile of all formulations met the API specification (see specification limits in the table) and therefore no degradation occurred during the preparation process.

Both API-containing batches showed low values in the API analysis, which results can be attributed to the small batch size as well as losses during preparation and samples used for IPC of the final mixture.

API-containing batches prepared by dry granulation with a weight ratio API/sugar of 1:2

All sugars previously investigated to prepare API/sugar tablets with a weight ratio of 1:5 using dry granulation were rated again at a ratio of 1: 2.

The preparation process is as described above. In this case, the resulting mixture was tabletted using an 8mm diameter punch.

Two types of disintegrants are used: microcrystalline cellulose as a standard disintegrant: (PH 112), and crosslinked polyvinylpyrrolidone used only for lots D001T/105For this batch, we are rightPH112 andthe use of (2) was investigated. Selection was made based on the cyclodextrin-based formulation previously prepared by dry granulation at an API/sugar ratio of 1:5 (see previous results)

Tables 29 and 30 summarize the composition of each API-containing formulation prepared by dry granulation at a weight ratio of API/sugar of 1:2, and the results of analytical tests performed on the final mixture and tablets. All API-containing batches showed suitable quality uniformity, API content uniformity and low impurity content. In most cases, the friability and hardness values meet specifications. In the case of batches D001T/093, D001T/095 and D001T/096, the results of the dissolution test on 6 tablets showed high RSD outside the specification values, the test extending to a sample of 12 tablets.

In two disintegrating agents: (PH112 and) The lower cyclodextrin-based tablets all showed good properties.

TABLE 29 Dry granulation-API/sugar weight ratio 1: 2. Composition and analytical results of the final mixture of API-containing batches.

API-containing batches prepared by direct compression were used at a weight ratio API/sugar of 1: 5.

Studies on saccharides having characteristics suitable for preparation by dry granulation using direct compression have led to the development of tablets having an API/sugar ratio of 1: 5.

Two types of disintegrants are used: microcrystalline cellulose as a standard disintegrant: (PH 112), and crosslinked polyvinylpyrrolidone used only for batch D001T/029

The preparation process comprises the following steps:

1. the API and excipients were weighed.

2. The stock was transferred to a double-layer polyethylene bag and mixed for about 5 minutes until a homogeneous powdered mixture was obtained.

3. The powdered mixture is transferred to the tablet press hopper.

4. The powdered mixture was compression molded using an eccentric tablet press equipped with a 10mm diameter punch.

The characteristics of the API-containing batches prepared using direct compression are shown in the table below.

TABLE 31 weight ratio of direct compression API/sugar 1: 5. Composition and analytical results of the final mixture of API-containing batches.

The results of the analytical tests are shown in Table 32.

TABLE 32 direct compression API/sugar weight ratio 1: 5. Analysis results of the tablets of the API-containing batches.

As shown in the above table, none of the tablets comprising API prepared by direct compression showed any critical difference from the tablets prepared by dry granulation, except that batch D001T/030 (filler: sucrose 97% + maltodextrin 3%) showed a non-uniform API content and a slightly increased friability value.

And (3) wet granulation:

placebo exploratory test

According to the results obtained in the first and second parts of the protocol, a wet granulation study was carried out on those sugars which were not suitable for dry granulation or direct compression.

The protocol for studying the wet granulation technique is shown below.

Each sugar was granulated according to the procedure described in the flow chart of figure 4. At the end of each step, the wet granulated sugar was dried and subjected to a compression moulding test to assess whether the granules were suitable for tabletting. Placebo batches were prepared only for granulated sugar for which compression molding trial results were suspect. The composition of the placebo trial and the associated analysis results are shown in table 33.

Placebo batches were prepared according to the following steps:

1. the sugars were wet granulated with water or sorbitol solution using a fluid bed or high shear granulator (see flow sheet and table 23 for the wet granulation preparation test above).

2. The wet granulated sugar is dried in a fluid bed granulator or in an oven.

3. The granulated sugar was sieved by using 850 micron and 710 micron sieves.

4. All ingredients of the respective formulations were weighed and mixed in a polyethylene bag for 2 minutes.

5. The powdered mixture was compression molded using an eccentric tablet press equipped with a 10mm diameter punch.

For all batches prepared, avicel ph112 and magnesium stearate were used as disintegrant and lubricant, respectively.

API-containing batches prepared by wet granulation with a weight ratio API/sugar of 1:5

All those saccharides that showed unsuitability for the preparation of tablets by dry granulation or direct compression techniques were subjected to manufacturing trials including the wet granulation process.

The preparation steps for these experiments performed on a laboratory scale are summarized below:

1. the sugars were wet granulated with water or sorbitol solution using a fluid bed or high shear granulator (see flow sheet and table 34 of the wet granulation manufacturing test above).

2. The wet granulated sugar is dried in a fluid bed granulator or in an oven.

3. Sieving was performed through 850 micron and 710 micron sieves.

4. The API and excipients were weighed and mixed in a double polyethylene bag for 5 minutes.

5. The powdered mixture was compression molded using an eccentric tablet press equipped with a 10mm diameter punch.

For all batches prepared, avicel ph112 and magnesium stearate were used as disintegrant and lubricant, respectively.

Tables 33 and 34 list the composition of each of the API-containing formulations prepared by wet granulation, and the results of the analytical tests performed on the final mixtures and tablets

In most cases, the results of the analytical tests carried out on the final mixture and on the finished product are in compliance with the specifications. No degradation occurred during the preparation process.

Of the sugars studied, only fructose ms (galem) is not suitable for processing by wet granulation: the friability of API-containing lot D001T/047 was high, while lot D001T/082 showed friability and hardness values outside specification.

Batches D001T/060, D001T/061, D001T/082 and D001T/086 had low values in the API analysis, whereas for batches D001T/082 and D001T/086, despite the use of 850 and 710 micron screens to screen the particles, the content uniformity was not met. This result may be attributed to poor powder mixing.

API-containing batches prepared by wet granulation with a weight ratio API/sugar of 1:2

All sugars previously investigated to prepare API/sugar tablets using wet granulation at a weight ratio of 1:5 were rated again at a ratio of 1: 2.

At a ratio of 1:2, fructose was not rated, as the resulting granules were not suitable for tableting.

For all batches prepared, avicel ph112 and magnesium stearate were used as disintegrant and lubricant, respectively.

To improve the uniformity of the API content, the API-containing batches were prepared using the following steps:

1. the sugar was wet granulated using the procedure previously optimized.

2. A mixture containing the API was prepared.

3. The mixture is subjected to dry granulation (billet preparation → billet screening).

4. The resulting mixture was tabletted with an 8mm diameter punch.

Step 3 (dry granulation of the mixture) is as described above.

Tables 36 and 37 show the composition and analytical results of API-containing batches prepared by using wet granulated sugar at an API/sugar weight ratio of 1: 2. In most cases, the friability is out of specification. The weight change of the API/sugar did not jeopardize the technical performance of batch D001T/084 (bulking agent: granulated mannitol).

API/mannitolInfluence of the alcohol weight ratio

Mannitol-based tablets were prepared to study the following API/mannitol ratio: 1:0.01, 1:0.1, 1:0.5, 1:1.7, 1:4, 1:5, 1:6, and 1: 10. Formulations with a weight ratio API/mannitol of 1:5 (standard formulations) were reported above.

For the preparation of these batches, avicel ph112 and magnesium stearate were used as disintegrant and lubricant, respectively. Regarding the preparation process, the wet granulated mannitol, bendamustine hydrochloride and excipients were precisely weighed for the ratios of 1:1.7, 1:4, and 1:6, and mixed in a double-layer polyethylene bag for 5 minutes. For batch D001T/110 (ratio 1: 10), premixing was performed. In this case, bendamustine hydrochloride was mixed with half the amount of the excipient mixture for 5 minutes. Thereafter, the resulting mixture was added to the remaining amount of excipient and mixed for another 5 minutes. The final blend was tabletted using a tablet press equipped with suitable punches (8 mm punches for 1:1, 1:1.7 and 1:2 ratios, 10mm punches for 1:4 and 1:6 ratios, 12mm punches for 1:7 ratios and 14mm punches for 1:10 ratios).

For the ratios of 1:0.01, 1:0.1 and 1:0.5, we used the above reported preparation procedure (wet granulation followed by dry granulation of the sugar) to improve the API content uniformity. The resulting mixture was tableted using a 6mm diameter punch.

The following tables (tables 38 and 39) summarize the compositions and analytical results of API-containing formulations prepared to study the effect of different API/mannitol ratios. Batches D001T/111, D001T/083 and D001T/106 showed high friability, whereas batches D001T/106, D001T/108 and D001T/109 did not meet the content uniformity, deviating from the trend of the data obtained before. This result may be attributed to the fact that these batches were prepared by using many novel bendamustine hydrochloride (cat # F08-05873) which may have different physical properties.

Carbohydrate combination study

Tables 40 and 41 show the results of the study on saccharide combinations.

The following combinations were studied:

mono-/disaccharide 1:1

（^*) Mannitol (Pearlitol 200 SD)/lactose anhydrous (SuperTab 21 AN)

Sorbitol (Neosorb P60W)/maltose (SunmaltS)

Oligosaccharide/monosaccharide 1:1

（^*) D-melezitose monohydrate/(^*) Anhydrous dextrose ST0.5

（^*) Granulated raffinose pentahydrate/(^*) Granulated mannitol (Pearlitol 200 SD)

Oligosaccharide/disaccharide 1:1

（^*) Granulated raffinose pentahydrate/lactose monohydrate (Supertab 14 SD)

Beta-cyclodextrin (KleposetC)/sucrose (EVSaccharide)

（^*) These sugars were granulated by wet granulation.

The preparation process comprises direct compression of the raw or granulated sugar.

These batches were prepared using avicel ph112 and magnesium stearate as disintegrants and lubricants, respectively, by performing the following steps:

1. the sugar (or granulated sugar), bendamustine hydrochloride, and excipients were accurately weighed and mixed in a double-layer polyethylene bag for 5 minutes.

2. The resulting mixture was tableted using a 10mm diameter punch.

Table 40. saccharide combination study. Composition and analytical results of the final mixture of API-containing batches.

Tablets prepared for studying the saccharide combination showed good properties overall. However, lots D001T/102 (raffinose pentahydrate/mannitol (Pearlitol 200 SD)) showed high friability, while lots D001T/100 and D001T/049 did not have uniform API content.

Example 22 Freeze-dried bendamustine hydrochloride (Ribomustin) and bendamustine hydrochloride/mannitol tablets (API/sugar weight ratio 1: 1.2)

Using commercially available preparations for intravenous administrationThe obtained lyophilizate, or wet granulated mannitol and bendamustine hydrochloride were used to prepare tablets containing bendamustine hydrochloride/mannitol in a weight ratio of 1: 1.2.

The preparation was carried out according to the following experimental procedure: freeze-drying the powder fromThe vial was removed and sieved using a 850 micron mesh. The resulting powder and lubricant (magnesium stearate) were accurately weighed and mixed in a polyethylene bag for 5 minutes. The mixture was slowly transferred into the compression chamber of a tablet press and manually compressed using an 8mm diameter punch to obtain a small billet. The billet was sieved using a 850 micron mesh and the resulting granules were hand pressed using an 8mm diameter punch.

Bendamustine hydrochloride/mannitol tablets were prepared using the same procedure as described above in this example.

The composition of the formulation is shown in table 42.

Table 42 Ribomustin and bendamustine/mannitol tablets. Composition of the final mixture of API-containing batches.

(. about.) 45.16% bendamustine hydrochloride and 54.20% mannitol

Table 43 shows data comparing tablets obtained using the lyophilized bendamustine hydrochloride/mannitol mixture to tablets obtained using the non-lyophilized bendamustine hydrochloride/mannitol mixture.

Table 43 Ribomustin and bendamustine/mannitol tablets. Analytical results of tablets containing API batches

With the impurity profile of bendamustine hydrochloride API as a reference indicator (see specification limits in the table), batch D001T/125 showed impurity HP1 to be out of specification. Dissolution test results highlighted that, although the dissolution behavior of the tablets containing the lyophilized bendamustine hydrochloride/mannitol mixture was faster after 10 minutes, the dissolution behavior of both formulations met the current specifications after 30 minutes. Lot D001T/126 was out of specification for friability and Lot D001T/125 was not tested due to the lack of sufficient amount of material.

Example 23: absolute bioavailability of oral bendamustine to cancer patients

A total of 12 patients were scheduled for a phase 1 open-labeled randomized, two-way crossover study to study the bioavailability of bendamustine following oral administration of a liquid-filled hard capsule formulation of bendamustine hydrochloride. 14 patients with multiple myeloma, B-cell type chronic lymphocytic leukemia or advanced indolent non-Hodgkin's lymphoma were recruited and treated with bendamustine. These patients were previously treated by intravenous bendamustine, but should be subjected to the last intravenous cycle (lasttrapenouscycle) at least seven days prior to the first administration of the study drug. After signing the informed consent and passing the screening period (day-21 to day-2), qualified patients were issued patient numbers specific to each clinical center. Patients received one of the following dosing regimens randomly on day 1, followed by another on day 8:

single oral dose of-110.2 mg (2X 55.1 mg) bendamustine hydrochloride

Single intravenous dose of-110 mg bendamustine hydrochloride

Bendamustine is provided in the form of a) an oral capsule (LFHC formulation (liquid-filled hard shell capsule)) and b) an intravenous solution (obtained after reconstitution of the powder used to prepare the injection solution). Utilizing 55.1mg bendamustine hydrochloride, 1.2mg methylparaben, 0.12mg propylparaben, 0.12mg top hydroxytoluene, 10.9mg ethanol, and 532.56mgRH40 LFHC formulations (per capsule) were prepared. The vial containing the powder for the concentrated solution is a product marketed in GermanyEach vial contained 100mg bendamustine hydrochloride and mannitol as an excipient. The product was reconstituted with water for injection as indicated in the package insert to give a final concentration of bendamustine hydrochloride of 2.5mg/ml and further diluted with 0.9% NaCl to about 500ml prior to administration to the patient.

Patients were admitted to the clinical center for phase 2 studies: day-1 to day 2 (stage 1) and day 7 to day 9 (stage 2). A total of 12 patients were randomized to receive treatment. 6 patients received such treatment: 110.2mg (2 × 55.1 mg) of single oral dose bendamustine hydrochloride (day 1), followed by a 100mg single intravenous dose of bendamustine hydrochloride (day 8); while the other 6 patients received treatment alternating the sequence described above. Between treatments, patients were subjected to an elution period of at least 7 days.

Bendamustine is metabolized by hydrolysis to the inactive metabolites monohydroxybendamustine (HP 1) and dihydroxybendamustine (HP 2), and by cytochrome P450 (CYP 1a 2) to the active metabolites γ -hydroxybenzbendamustine (M3) and N-desmethyl bendamustine (M4).

Plasma and urine samples were tested for concentrations of bendamustine and bendamustine active metabolites (M3 and M4) on days 1 and 8 after oral and intravenous administration of bendamustine. After completion of the second treatment period or after early replacement/withdrawal, patients were returned to the clinical center for a study end follow-up (end-of-studyvisit) of 7 to 14 days. Thereafter, pharmacokinetic parameters of bendamustine and its metabolites were calculated.

No interim analysis was scheduled or performed.

The following results were obtained:

the crowd:

of the 23 patients screened for this study, 14 patients were randomized to treatment and received at least 1 dose of study drug. This includes: 6 patients received treatment in an oral/intravenous order and 8 patients received treatment in an intravenous/oral order. In these 14 patients:

position-1 was excluded due to violation of protocol (combination) and receiving only oral medication and therefore not administered intravenously;

position 1 was excluded from oral analysis due to vomiting without meeting the requirements for bioavailability evaluation;

position-1 was excluded from intravenous administration due to adverse event (adverevent). The patient received only oral administration and not intravenous administration.

10 of the 14 patients (71%) were male and all were caucasian. Patients ranged in age from 54 to 82 years, with an average of approximately 70 years. Of these patients 7 had multiple myeloma, 4 had indolent non-hodgkin's lymphoma, and 3 had chronic lymphocytic leukemia.

Pharmacokinetic results:

plasma pharmacokinetic parameters for bendamustine (base), M3, and M4 are shown in table 44, table 45, and table 46, respectively. Absolute bioavailability (oral AUC) of bendamustine according to statistical analysis_infAnd venous AUC_infRatio of (d)) was 66% (geometric mean: 90%, CI: 55%, 78%). C after oral administration_maxIs after intravenous administration C_max42% (90% CI: 32%, 54%).

Table 44: plasma pharmacokinetic parameters of bendamustine

Note:

all patients received at least 1 dose of study drug with sufficient plasma concentration data to obtain at least 1 pharmacokinetic parameter, except 1 patient who was not confident in its pharmacokinetic data due to vomiting AE (altered pharmacokinetic analysis settings).

The arithmetic mean. The geometric mean was 66% (90% CI: 55%, 78%).

C after oral administration_maxIs after intravenous administration C_max42% (90% CI: 32%, 54%).

Table 45: plasma pharmacokinetic parameters of M3

Note:

all patients received at least 1 dose of study drug with sufficient plasma concentration data to obtain at least 1 pharmacokinetic parameter, except 1 patient who was not confident in its pharmacokinetic data due to vomiting AE (altered pharmacokinetic analysis settings).

Table 46: plasma pharmacokinetic parameters of M4

Note:

all patients received at least 1 dose of study drug with sufficient plasma concentration data to obtain at least 1 pharmacokinetic parameter, except 1 patient who was not confident in its pharmacokinetic data due to vomiting AE (altered pharmacokinetic analysis settings).

Absorption of bendamustine after oral administration_maxAt about 0.95 hours, the individual values were between 15 minutes and 1.8 hours. After intravenous administration, the mean CL was 21.2L/h. Mean t after oral administration and after intravenous administration_1/2All for about 30 minutes. After intravenous administration, V_ZAnd V_SS14.7L and 10.3L respectively.

Plasma exposure of M3 and M4 was significantly lower than bendamustine. Mean AUC of bendamustine after oral administration_inf10.6 times and 88 times higher than M3 and M4, respectively. AUC of M3 for oral and intravenous administration compared to bendamustine_infValue and AUC of M4_infThe values are all similar. AUC of M3 after oral administration according to statistical analysis_infIs AUC after intravenous administration_inf86% (90% CI: 76%, 98%). 88% in the case of M4 (90% CI: 77%, 102%).

The urine pharmacokinetic parameters for bendamustine, M3, and M4 are shown in tables 47, 48, and 49, respectively. The percentage of dose expelled in the unaltered urine (urineexchangeable) was low (2.6% and 2.1% for oral and intravenous administration, respectively).

Table 47: urine pharmacokinetic parameters of bendamustine

Note:

all patients received at least 1 dose of study drug with sufficient plasma concentration data to obtain at least 1 pharmacokinetic parameter, except 1 patient who was not confident in its pharmacokinetic data due to vomiting AE (altered pharmacokinetic analysis settings).

Table 48: urine pharmacokinetic parameters of M3

Note:

all patients received at least 1 dose of study drug with sufficient plasma concentration data to obtain at least 1 pharmacokinetic parameter, except 1 patient who was not confident in its pharmacokinetic data due to vomiting AE (altered pharmacokinetic analysis settings).

Table 49: urine pharmacokinetic parameters of M4

Note:

all patients received at least 1 dose of study drug with sufficient plasma concentration data to obtain at least 1 pharmacokinetic parameter, except 1 patient who was not confident in its pharmacokinetic data due to vomiting AE (altered pharmacokinetic analysis settings).

Safety results:

bendamustine is safe and well tolerated both orally and intravenously. Overall, 6 patients (43%) experienced adverse events during treatment in oral treatment and 3 patients (25%) experienced adverse events during treatment in intravenous treatment. 4 patients receiving oral administration (29%) and 0 patients receiving intravenous administration experienced at least 1 adverse event recognized by the investigator as being related to study drug; these adverse events include: headache in 1 patient, headache and fatigue in 1 patient, nausea in 1 patient, and vomiting in 1 patient. Except for emesis (grade 2 severity), these events were grade 1 in severity.

Most adverse events were of grade 1 or grade 2 in severity. Grade 3 adverse events (increased serum creatinine, hypokalemia and acute renal failure) and grade 4 adverse events (thrombocytopenia) occurred in 1 patient receiving oral administration, both of which were considered by the investigators to be associated with multiple myeloma in the patients and not with study medication. Serum creatinine increases and acute renal failure are serious adverse events leading patients to early discontinuation of the study. No mortality events occurred during the study.

No clinically significant trends were observed in mean changes at baseline or absolute changes for hematological, biochemical, urinalysis or vital parameters. A few patients had abnormal hematological or biochemical findings (reported as adverse events); the investigators considered this to be independent of study drug.

The mean change from baseline was small in heart rhythm and similar between treatment groups. Since patients in this study had different ages and medical history, most patients had at least 1 "abnormal, clinically insignificant" electrocardiogram result in the screening and/or study. During screening and subsequent intravenous and oral dosing, 1 patient in the intravenous/oral group was observed to have abnormal, clinically significant atrial fibrillation, non-specific ST-segment depression, and left off-cardiac axis.

And (4) conclusion:

the absolute bioavailability of bendamustine after a single oral administration using the capsule was 66% (geometric mean; 90% CI: 55%, 78%).

Bendamustine has an average CL, Vz, and Vss of 21.2L/h, 14.7L, and 10.3L, respectively, after intravenous administration.

Bendamustine is rapidly absorbed after oral administration (t)_maxMedian of about 0.95 hours). Average t_1/2About 30 minutes. After oral use, approximately 2.6% of the dose is excreted in intact urine, whereas in the case of M3 0.6% is excreted, in the case of M4 0.1%. After oral administration, the exposure of M3 and M4 was about 9% and 1% of bendamustine, respectively.

Based on reported adverse events, clinical laboratory evaluations, vital signs, physical examinations and electrocardiograms, single dose oral (110.2 mg) and intravenous (100 mg) formulations of bendamustine were shown to be safe and well tolerated in most populations of patients with indolent non-hodgkin's lymphoma, multiple myeloma or B-cell type chronic lymphocytic leukemia for more than middle-aged years.

Industrial applications

The pharmaceutical compositions of the present invention exhibit a number of advantages. The patient can easily use the pharmaceutical composition without the help of supervising medical staff. It may no longer be necessary to go through the time-consuming procedure of hospital visits, thereby improving patient compliance.

Since the dosage form is solid, it can be swallowed, which means that the patient does not need to wait until the active ingredient is dissolved. In addition, since the formulation has good stability, the formulation can be easily stored at room temperature and does not require any special storage conditions.

By using the dosage form of the present invention, the volume of the dosage form can be significantly reduced. The reduction in size is advantageous both from a production and handling perspective and from a patient compliance perspective.

The pharmaceutical compositions exhibit high in vitro dissolution rates which reduce bendamustine degradation in vivo, thereby resulting in improved in vivo bioavailability of bendamustine.

Claims (15)

1. A pharmaceutical composition in oral dosage form comprising: bendamustine hydrochloride as an active ingredient and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient is selected from the group consisting of:

-polyethylene glycol glycerol hydroxystearate;

-polyoxyethylene-35-castor oil, or a combination of polyoxyethylene-35-castor oil and lauroyl macrogolglycerides, wherein the concentration of the lauroyl macrogolglycerides in the composition is from 10% to 50%;

ethylene oxidePropylene oxide block copolymerL44NF, orL44NF in combination with colloidal silica, wherein the concentration of the colloidal silica in the composition is from 2% to 8%, orA combination of L44NF and lauroyl macrogolglycerides, wherein the concentration of the lauroyl macrogolglycerides in the composition is from 10% to 45.4%;

wherein the composition exhibits a bendamustine dissolution behavior when measured according to the european pharmacopoeia in 500ml of dissolution medium at a pH of 1.5 using a paddle apparatus at a rotation speed of 50 rpm: at least 60% dissolved in 10 minutes, at least 70% dissolved in 20 minutes, and at least 80% dissolved in 30 minutes.

2. Pharmaceutical composition according to claim 1, characterized in that it comprises from 10mg to 1000mg of said active ingredient.

3. Pharmaceutical composition according to any one of the preceding claims, characterized in that it is contained in a hard gelatin capsule.

4. Use of a pharmaceutical composition according to any one of the preceding claims for the preparation of a medicament for the oral treatment of a medical condition selected from: chronic lymphocytic leukemia, acute lymphocytic leukemia, chronic myelocytic leukemia, acute myelocytic leukemia, hodgkin's disease, non-hodgkin's lymphoma, multiple myeloma, breast cancer, ovarian cancer, small cell lung cancer, and non-small cell lung cancer.

5. The use of claim 4, wherein the pharmaceutical composition is used in combination with at least one further active agent, wherein the use of the at least one further active agent is performed before, simultaneously with or after the use of the pharmaceutical composition and the further active agent is selected from the group consisting of a CD 20-specific antibody, an anthracycline derivative, vinblastine or a platinum derivative.

6. Use according to claim 5, characterized in that: the CD20 specific antibody is rituximab; the anthracycline derivative is adriamycin or daunorubicin; the vinblastine is vincristine, and the platinum derivative is cisplatin or carboplatin.

7. Use according to any one of claims 4 to 6, in combination with at least one corticosteroid, wherein said corticosteroid is administered before, simultaneously with or after the administration of said pharmaceutical composition.

8. Use according to claim 7, characterized in that: the corticosteroid is prednisone or prednisolone.

9. The use according to any one of claims 4 to 6, wherein the active ingredient is administered at 50mg/m per course of treatment²Per person to 1000mg/m²Dose administration between/each person.

10. Use according to any one of claims 4 to 6, wherein the dosing regimen comprises at least:

-administration of a dose of 100mg/m on days 1 and 2²Per person to 600mg/m²Bendamustine per human being,

-optionally, on days 1 to 5,the dosage of intravenous administration or oral administration is 50mg/m²To 150mg/m²A corticosteroid of (a), and

-optionally administering an appropriate dose of an additional active agent selected from the group consisting of a CD 20-specific antibody, an anthracycline derivative, vinblastine or a platinum derivative; and

after 2-4 weeks apart, the dosing regimen was repeated 4 to 15 times.

11. The use according to any one of claims 4 to 6, wherein the active ingredient bendamustine is administered in a dosing regimen selected from the following regimens:

200-300mg on day 1 and day 2, optionally followed by a maintenance dose of 50mg, once a day,

-from day 1 to day 14, including day 14, administering 50mg daily, or

Administration of 150mg once a week for 3 weeks.

12. The use according to any one of claims 4 to 6, wherein the patient has non-Hodgkin's lymphoma and the dosing regimen comprises: the total amount of active ingredient bendamustine was 200 mg/person/day from day 1 to day 5, 2mg vincristine was administered intravenously from day 1, and 100mg/m was administered intravenously from day 1 to day 5²Prednisone, repeating the treatment every three weeks until the non-hodgkin's lymphoma is ameliorated.

13. The use according to any one of claims 4 to 6, wherein the patient has multiple myeloma and the dosing regimen comprises: the dosage on the 1 st day and the 2 nd day is 100-250mg/m²Bendamustine hydrochloride at body surface area, and administered 60mg/m intravenously or orally from day 1 to day 4²Prednisone, repeating the treatment every four weeks until the multiple myeloma is improved.

14. Use according to any one of claims 4 to 6, wherein the patient is suffering from chronic lymphocytic leukemia and the dosing regimen comprises: the dosage on the 1 st day and the 2 nd day is 100-200mg/m²Bendamustine hydrochloride at body surface area, and administered 60mg/m intravenously or orally from day 1 to day 4²Said treatment is repeated every four weeks until said chronic lymphocytic leukemia is ameliorated.

15. The use of claim 10, wherein the patient has follicular, indolent or mantle cell lymphoma and the dosing regimen comprises: the dosage of the drug administered on day 1 is 375mg/m²And orally additionally administered 100mg/m on days 1 and 2²To 200mg/m²Is repeated every 28 days until each of said lymphomas is ameliorated.