UA81304C2 - Formulation of cladribine (variants), process for the preparation thereof, cladribine-cyclodextrin complex and use thereof - Google Patents

Abstract

Abstract of the disclosure provided are compositions of cladribine and cyclodextrin which are especially suited for the oral administration of cladribine.

Description

Description of the invention

The present invention relates to a composition including a cladribine-cyclodextrin complex, formulated 2 in a solid oral dosage form and to a method of increasing the oral bioavailability of cladribine.

Cladribine, which is an acid-labile drug, has a chemical structure that is presented below:

MN

2 sh M, g u, a nNOSN»o s (8)

Fee

This compound is also known as 2-chloro-2-deoxyadenosine or 2-SdaDA. Cladribine is a white, non-hygroscopic, crystalline powder that includes individual crystals and crystalline aggregates.

Cladribine is an antimetabolite used in the treatment of lymphoproliferative disorders. "-

It has been used to treat experimental leukemias, such as I 1210, and clinically for squamous cell leukemia and chronic lymphocytic leukemia, as well as Waldenstrom's macroglobulinemia. It has also been used as an immunosuppressant and as a treatment for many autoimmune conditions, including rheumatoid arthritis, inflammatory bowel disease (eg, Crohn's disease, ulcerative colitis), and multiple sclerosis |see, e.g., U. I. PshetagKk, Siip. RNpaptasokKipei, 32(2): 120-131, 1997). It has also been investigated, either experimentally or clinically, in the treatment of, for example, lymphoma, cell histiocytosis -o

Langerhans, lupus erythematosus, chronic plaque psoriasis, César syndrome (Zegagu zupagote), Bing-Neel syndrome (Vipd-Mee! zupagote), recurrent glioma and solid tumors. Oral delivery of drugs is usually preferable to parenteral delivery for many reasons, such as primarily patient contact, or for cost or therapeutic reasons. Contact

The patient is improved because oral dosage forms reduce the number of repeated visits to health care facilities, or the discomfort of injections or the long infusion time associated with some active drugs. At a time when health care costs are constantly increasing, the reduced costs associated with oral administration compared to parenteral administration are of great importance. The cost of parenteral administration is significantly higher due to the requirement that a healthcare professional administer cladribine in a healthcare setting, which also includes all (“in”) associated costs associated with such administration. In addition, in some cases, therapeutic considerations are as follows

T" as the need for slow release of cladribine over a long period of time can practically only be achieved by oral or transmucosal delivery.

However, until now, oral delivery of cladribine has been unacceptable due to low bioavailability | see, for example, U. Shiietagk et al., U. Siipy. Opsoi, 10(10): 1514-1518, 1992), and suboptimal interpatient variability (see, for example, .). And PShetagK, Siip. Rpaptasokipei 32 (2): 120-131, 1997). See also, A. (F) TagaztsiK, and others, describing low absorption and pH-dependent instability (Agsy. Ittipoi, ei Tnegariae

Ge Ehreg., 42: 13-15, 1994))|.

Cyclodextrins are cyclic oligosaccharides consisting of cyclic o-(1 4) linked in O-glucopyranose units. Cyclodextrins with six to eight units are designated as α-, β- and γ-cyclodextrin, respectively. The number of units determines the size of the cone-shaped cavity that characterizes cyclodextrins and into which drugs can be incorporated to form stable complexes. Many derivatives of o-, D- and y-"-cyclodextrins are known, in which one or more hydroxyl groups is/are replaced by ether groups or other radicals. Thus, these compounds are known complexing agents and 65 were previously used in the field of pharmaceuticals for the formation inclusion complexes with water-insoluble drugs and, thus, for their dissolution in an aqueous medium.

Recently, Zspcy2 et al., (U.S. Patent Mob,194,395 B1), described the complexation and solubilization of cladribine with cyclodextrin. The Zspci et al. patent primarily addresses the problems inherent in previously described aqueous formulations of cladribine, particularly for subcutaneous and intramuscular administration. Zepa et al. found that cladribine is not only significantly more soluble in aqueous media when formulated with cyclodextrin, but is also more resistant to acid-catalyzed hydrolysis when combined with cyclodextrin.The latter fact has been found to be of some benefit in the formulation of solid oral dosage forms because the compound is usually hydrolyzed at the acidic pH of the gastric contents. Zspci: et al do not appear to describe any actual work in connection with solid oral 7/0 dosage forms. In fact, they only describe a process for preparing solid dosage forms, which is a melt extrusion process in which cladribine and cyclodextrin are mixed with other optional additives and then heated until melting occurs.In addition, the wide dosing and the ranges of 1 mg to 15 mg cladribine and 100 mg to 500 mg cyclodextrin listed in this patent do not provide any precision as to what specific amount of cyclodextrin is present with a given amount of cladribine in a solid/5 oral dosage form. Indeed, these dosage ranges include many combinations that may be acceptable as mixtures but not acceptable for complex formation. For example, the ratio of mg cladribine to 500 mg cyclodextrin contains too much cyclodextrin, as a result of which the drug would not easily leave the complex and achieve its therapeutic function. On the other hand, 15 mg of cladribine and only 10 mg of cyclodextrin would not be sufficient to complex this amount of cladribine.

The patent of Zsptia2 et al suggested improving the stability of cladribine in oral dosage forms by combining/complexing it with cyclodextrin, but did not describe improving the oral bioavailability of the drug due to this possibility; in fact, this patent does not describe or suggest a method for increasing or maximizing the bioavailability of cladribine from a solid oral dosage form of cladribine and cyclodextrin, or a composition specifically designed to do so. with

Many researchers have studied the solubility of specific drugs in water containing different concentrations of selected cyclodextrins in order to demonstrate that increasing concentrations of i) cyclodextrins increase the solubility of drugs at selected temperatures and pH levels, such as described in the patent Zspics2 et al. Studies by the phase solubility method have also been carried out by various researchers in order to explain the nature of the formation of complexes, for example, whether they form "E

Zo Cyclodextrin and drug 1:11 complex or 1:2 complex; |see, e.g., Nagada et al., Pat

USA Mo4,497,803), which refers to the inclusion complexes of antibiotics of the lankacidin group with cyclodextrin, and Zpipod and others (US Patent Mo4,478,995), which refers to the complex of the addition salt of acid (2'-benzyloxycarbonyl)phenyl-trans- 4-guanidinomethyl-cyclohexanecarboxylate with cyclodextrin.

While Zspcii7 et al describe that a cladribine-cyclodextrin complex improves the aqueous solubility and acid stability of cladribine, the prior art does not suggest how to maximize or enhance the beneficial effects of complexation on bioavailability and interpatient variability when the complex must be administered. in a solid oral dosage form.

To date, it has been found that amorphous cyclodextrins can be combined with cladribine to form a particularly superior product that can be incorporated into a solid oral dosage form. This product is a cladribine-cyclodextrin complex, and a solid oral dosage form containing it improves oral bioavailability and/or achieves lower interpatient and/or intrapatient variability of the drug. with This invention provides a complex cladribine-cyclodextrin complex, which is a homogeneous amorphous mixture of (a) an amorphous inclusion complex of cladribine with amorphous cyclodextrin and (B) an amorphous

Free cladribine bound to amorphous cyclodextrin as a non-inclusion complex, and a pharmaceutical composition including said complex formulated in a solid oral dosage form. Thus, cyclodextrin itself is amorphous, the inclusion complex with cladribine is amorphous (and is mostly saturated with cladribine), and free cladribine, which forms a non-inclusion complex, is amorphous. 2) The present invention also provides a method of increasing or improving the oral bioavailability of cladribine, which includes oral administration to a patient in need of such treatment, a pharmaceutical composition, which includes a complex cladribine-cyclodextrin complex, which is a homogeneous amorphous mixture (a) and" amorphous inclusion complex of cladribine with amorphous cyclodextrin and (B) amorphous free cladribine bound to amorphous cyclodextrin as a non-inclusion complex formulated in a solid oral dosage form that maximizes the amount of cladribine in inclusion and non-inclusion complexes.

In addition, the present invention provides for the treatment of conditions responsive to the use of cladribine in mammals by applying to them the composition of the present invention. Also provided is the use of cladribine in (F) the preparation of pharmaceutical compositions of the invention for use in the treatment of cladribine-sensitive conditions and for increasing the oral bioavailability of cladribine.

Moreover, the present invention provides a method of obtaining a complex cladribine-cyclodextrin complex, which includes the steps of: () combining cladribine and amorphous cyclodextrin in water at a temperature of from about 40 to about 802C and maintaining said temperature for a period of from about 6 to about 24 hours; (i) cooling the obtained aqueous solution to room temperature; and (iii) lyophilization of the cooled solution to obtain an amorphous product. 65 In yet another additional aspect, the present invention provides a pharmaceutical composition that can be prepared by a method that includes the steps of:

() combining cladribine and amorphous cyclodextrin in water at a temperature of about 40 to about 80°C and maintaining said temperature for a period of about 6 to about 24 hours; (iii) cooling the obtained aqueous solution to room temperature; (ii) lyophilization of the cooled solution to obtain an amorphous product; and (im) formulating the amorphous product into a solid oral dosage form.

A more complete appreciation of the present invention and its many attendant advantages will be readily understood by reference to the following detailed description and the accompanying drawing, wherein this Figure is a graphical representation of the results of a phase solubility study, which is plotted against various molar 70 concentrations of hydroxypropyl-P-cyclodextrin (HRDSO ) from different molar concentrations of cladribine, while the (-) sign is the data processing results obtained for complexation under the conditions specified in Example 2, presented below.

The following definitions and basic wording are applicable throughout this specification and paragraphs

Formulas of the invention.

The patents, published applications, and scientific literature referenced in this specification establish the knowledge of those skilled in the art and are incorporated herein by reference in their entirety to the same extent as if each were specifically and individually shown. embedded as a link. Any conflict between each reference cited in this specification and the specific teachings of this specification shall be resolved in favor of the latter. Moreover, any conflict between the prior art definition of a word or phrase and the definition of a word or phrase specifically provided in this specification will be resolved in favor of the latter.

The term "inclusion complex", as used herein, refers to a complex of cladribine with a selected cyclodextrin, where the hydrophobic part of the cladribine molecule (nitrogen-containing ring system) is inserted into the hydrophobic cavity of the cyclodextrin molecule. More often, such an inclusion complex is simply referred to as the HaCcyclodextrin complex of the medicinal product.

The term "non-inclusion complex" refers to a complex that is not an inclusion complex; instead of the hydrophobic part of cladribin being inserted into the cavity of cyclodextrin, the non-inclusion complex is formed mainly with the help of hydrogen bonds of hydroxyl groups and amino groups on "free" cladribin (i.e., cladribin is not in the inclusion complex) to hydroxyl groups on the outer "And the surfaces of the cyclodextrin toroidal structure (for example, in the case of hydroxypropyl-D-cyclodextrin, the hydroxypropyl and hydroxyl groups on the glucose ring). This is a more loosely bound association of molecules than an inclusion complex. Go)

As used in this specification, in an interlocutory phrase or in the body of a Claim clause, the terms "comprising" and "comprising" should be interpreted as having a non-limiting meaning. That is, the terms - should be interpreted synonymously with the phrases "having(s), at least" or "which include(s), at least" When c is used in the context of a method description, the term "including" means that the method includes, at least, stages, but may also include additional stages. When this term is used in the context of describing a composition, the term "comprising" means that the composition includes at least the listed elements or components, but may also include additional elements or components.

The terms "consisting mainly of" or "consisting mainly of" have a partially limited - c meaning, that is, they do not allow the inclusion of stages or elements or components that would significantly change the main characteristics of the method or composition; for example, stages or elements or components that would significantly "" affect the desired properties of the compositions described in this specification, that is, the method or composition is limited to certain stages or materials and those that do not significantly affect the basic and novel characteristics of the present invention. and novel features in this specification are the provision of a complex (ee) cladribine-cyclodextrin complex which is a homogeneous amorphous mixture of (a) an amorphous complex - inclusion of cladribine with amorphous cyclodextrin and (b) amorphous free cladribine bound to amorphous cyclodextrin as non-inclusion complex formulated into a solid oral dosage form to (95) provide improved bioavailability and/or lower inter-patient variability and/or greater than 50 lower intra-patient variability after administration.Important to the invention is the combination of the amorphous nature of the starting cyclodextrin and the level solubility in water showing cladribine (approximately 5 mg/ml at c imnate c" temperature), and then its ability to form a hydrogen bond, which can be used under certain conditions described below, and which allows you to get a specific amorphous mixture that is clearly well suited for optimizing the oral bioavailability of cladribine. 5B The terms "consisting of" and "consisting of" are limited terminology and allow only the enumerated stages or elements or components to be included. o As used in this description of the invention, the singular forms specifically also include the plural forms of the terms to which they refer, unless the context clearly indicates otherwise.

The term "approximately" is used herein to mean near, in the vicinity of, approximately, or near. 60 When the term "approximately" is used in conjunction with numerical values, it modifies that range by extending the limits above and below the specified numerical values. Generally, the term "about" or "approximately" is used herein to correct a numerical value above and below the set value by a deviation of 20905.

The term "amorphous" as used herein refers to a non-crystalline solid. 65 The cyclodextrins included in this description are themselves amorphous because each of them consists of a plurality of individual isomers, and their complexes with cladribine are also amorphous. In addition, complexation conditions can be selected (increased temperature and prolonged complexation time, as described below) such that a supersaturated solution of cladribine is formed. After cooling, due to the amorphous nature of the complex and cyclodextrin, some excess free cladribine does not precipitate, but is rather trapped in an amorphous form in a homogeneous mixture with the (predominantly saturated) amorphous cladribine-cyclodextrin inclusion complex. This excess of cladribine forms a more weakly bound molecular association, or non-inclusion complex, with cyclodextrin by means of hydrogen bonds. This then further increases the amount of cladribine in the product; this additional cladribine, because it is amorphous and also because it forms a homogeneous mixture with the amorphous inclusion complex, is expected to be partially protected from 7/0 decomposition by gastric acid (although it may not be as well protected as cladribine, which is in the form of an inclusion complex).

The term "saturated", when used in conjunction with a cladribine complex in amorphous cyclodextrin, means that the complex is saturated with cladribine, i.e., the complex contains the maximum amount of cladribine that can be complexed (by both inclusion complex formation and /5 non-inclusion) with a given amount of cyclodextrin under the complexation conditions used. A phase solubility study can be used to obtain this information, as described in more detail below. (Conditions for complexation are also described in more detail below). Alternatively, a saturated complex can be achieved empirically by simply adding cladribine to an aqueous solution of a selected cyclodextrin until a precipitate (uncomplexed cladribine) forms; it is clear that excess cladribine, if present, is removed (by filtration or centrifugation) and the solution is lyophilized to obtain a dry saturated complex.

The expression "mainly" as in "mainly free" means within 2095 of a precisely calculated amount, preferably within 1095, more preferably within 5905.

The term "interpatient variability" refers to the variability among patients to whom a medicinal product is administered. The term "intrapatient variability" refers to the variability experienced by an individual patient when receiving doses of the drug at different times.

As used in this specification, the recitation of a numerical series for a variable value is intended to convey that the invention may be practiced with variable values equal to any value in that series. Thus, for a variable that is essentially discrete, the variable value can be equal to any whole "g" value of a numerical series that includes the endpoints of the series. Similarly, for a variable that is essentially continuous, the variable can be equal to any real value of a numerical series that includes the endpoints of the series. at

As an example, a variable that is described as having a value between 0 and 2 may be 0, 1, or 2 for c variables that are essentially discrete, and may be 0.0, 0.1, 0.01, 0.001, or any other valid value for variables that are essentially continuous. --

In the description of the invention and the clauses of the Claims of the Invention, the singular forms include the plural forms of the terms, unless the context clearly indicates otherwise. As used in this specification, unless specifically indicated otherwise, the word "or" is used in the "inclusive" sense of "and/or" and in the "non-inclusive" sense of "either/or".

Technical and scientific terms used in this specification have the meanings conventionally understood by one skilled in the art to which the present invention relates, unless otherwise indicated. "

Reference is made in this description to various methods and materials known to one skilled in the art. The standard works referred to which teach the basic principles of pharmacology include those of Soodtap and Sitap Te Riagtasoiodis! Vaziz Oi Tpegaretsiisv. 1072 edition, Meogham/ NIII Sotrapiez Ips., "» Mem Hogk (2001).

Detailed references are made below to specific embodiments of the present invention. Although the present invention will be described in conjunction with these specific embodiments, it should be understood that they do not limit the present invention to (ee) such specific embodiments. Rather, they are intended to cover variants, modifications, and equivalents that may be included within the spirit and scope of the present invention as defined in the appended Claims. In the following description, numerous specific details are set forth in order to provide a comprehensive understanding of the present invention. The present invention may be practiced without some or all of these specific details. In other cases, well-known process operations are not described in detail in order not to unnecessarily complicate the understanding of the essence of the present invention.

Chl» This invention provides compositions, as well as methods of preparation and use of pharmaceutical compositions, useful for achieving desired pharmacokinetic properties. Such compositions derive from the discovery that solutions of cyclodextrin and cladribine in which cladribine is in its highest thermodynamic state, when applied to the gastric mucosa through which they are absorbed, are associated with improved absorption of cladribine, reflected by higher bioavailability and/or lower

F, interpatient variability. It will be assumed, without attempting to limit the present invention in this way, that upon dissolution (for example, by contact with a liquid, such as a body fluid), the dry compositions according to the invention form a locally saturated solution of cladribine in which cladribine is in a state of highest thermodynamic activity ( NTA), thereby promoting absorption. Cladribine has a sufficiently low, although it cannot be said insignificant, inherent solubility in water; in fact, it is partially soluble in water. Free cladribine, formed from the decomposition of these inclusion or non-inclusion complexes in a saturated aqueous solution, attempts to achieve a more stable level of activity by absorption through the gastric mucosa. 65 In view of the above, it is apparent that in order to create optimal pharmaceutical compositions, in a solid oral dosage form, these dosage forms should be formulated to release a localized saturated solution of cladribine, upon contact of these solid dosage forms with body fluid at the mucosa in which cladribine is in its NTA state. In order to provide such a localized saturated solution, it is important to first determine the optimal ratio of cladribine to amorphous cyclodextrin, referred to herein as the NTA ratio, for use in a solid dosage form.

The HTA ratio is established empirically and is defined as the ratio of cladribine to amorphous cyclodextrin, which corresponds to the maximum amount of cladribine that can be complexed with a given amount of cyclodextrin. The HTA ratio can be determined using an empirical method, such as a 7/0 phase solubility study to determine the saturation concentration of cladribine, which can be solubilized using amorphous cyclodextrin solutions of various concentrations. Therefore, this method establishes the concentrations at which a saturated cladribine-cyclodextrin complex is formed. It is noted that the molar ratio, represented by a point on the phase solubility curve, shows how many moles of amorphous cyclodextrin are the minimum required to keep the drug in the complex, under /5 given conditions; this ratio can then be converted to a mass ratio. For example, if the phase solubility diagram shows that 9 moles of a given cyclodextrin are required to hold cladribine in a saturated complex, then multiplying the number of moles of cladribine by its molecular weight and multiplying the number of moles of the selected cyclodextrin by its molecular weight can arrive at a product ratio that is acceptably optimized mass ratio. The study of phase solubility also provides information about the nature of the formed cladribine-cyclodextrin complex, for example, whether the complex is a 1:1 complex (1 molecule of the drug is complexed with 1 molecule of cyclodextrin) or a 1:2 complex (1 molecule of the drug is complexed with 2 molecules cyclodextrin).

In accordance with the present invention, determinations can be initiated using either a selected amorphous cyclodextrin, such as hydroxypropyl-D-cyclodextrin (HDRSO) or hydroxypropyl-y-cyclodextrin, or cladribine as a fixed variable to which excess of the other is added to determine various points. solubility graph (showing saturated cladribine-cyclodextrin complexes), and draw the resulting line. at

Similarly, cladribine is added to an aqueous solution having a known concentration of amorphous cyclodextrin under conditions found empirically to promote complex formation. Generally, the complexation is carried out with heating at about 4522 to about 602C for a significant period of time, for example at least 6-9 hours; it is believed that even better results can be obtained by heating to about 802 for 24 hours. Excess precipitated cladribine is then removed and the concentration of cladribine is further measured. The measured concentration represents the amount of cladribine that is dissolved by a given concentration of amorphous cyclodextrin. This method is repeated for various known concentrations of cyclodextrin until several points of the curve are obtained. Each point of the curve represents the concentration of cladribine dissolved in selected amorphous cyclodextrin of a known concentration. These curve points are then plotted to (se) show the concentration of cladribine in relation to the various concentrations of cyclodextrin used. This curve represents a phase solubility diagram that can be used to determine the amount of cladribine for any specific concentration of cyclodextrin used to form a solution under a given set of complexation conditions. It is understood that the water solubility of cladribine is approximately 70 Bmg/mL at room temperature and will be higher at elevated temperatures. Therefore, the points of the curve correspond to - the amount of cladribine dissolved in an aqueous solution of NRDSO or other amorphous cyclodextrin under the selected conditions; then, after lyophilization of the solution, a complex cladribin-cyclodextrin complex is obtained, which is a homogeneous amorphous mixture of (a) an amorphous inclusion complex of cladribin with amorphous cyclodextrin and (b) amorphous free cladribin bound to amorphous cyclodextrin as a non-inclusion complex. If equilibrium conditions are reached during the complexation process, the amorphous (ee) cladribine-cyclodextrin complex will be saturated with cladribine. - One skilled in the art will be able to appreciate that the concentrations at which saturated complexes of cladribine with amorphous cyclodextrin are formed (and thus also the HTA ratio), (95) can be determined by various alternative methods. Thus, any method known in the art that is acceptable for determining these concentrations is within the scope of this invention.

It was found that the desired pharmacological properties (improved bioavailability and/or coefficient of variation in

I" compared to traditional approaches) associated with mixtures of inclusion complexes and non-inclusion complexes of cladribine and cyclodextrin.

Using essentially amorphous cyclodextrins, for example, hydroxypropyl-dD-cyclodextrin, oo hydroxypropyl-y-cyclodextrin, arbitrarily methylated cyclodextrins, and the like, with cladribine, which is partly

GF! a water-soluble compound (capable of forming H-bonds with its free hydroxyl and amino groups), cladribine provides increased solubility in solutions of these cyclodextrins. This gives not only increased solubility in water, but also H-bonded association of cladribine molecules with cyclodextrin, separately from the active material of the formed inclusion complex. for a person skilled in the art will understand that the phase solubility diagram for each given concentration ratio is the starting point of his study, based on which variables (reagents, concentrations, temperature, and time) can be varied to improve the association of inclusion complex molecules and of the non-inclusion complex, contributing to a higher or lower proportion of the other type of association in the final product. Deviations from the ratio of cladribine to cyclodextrin, temperature, and/or dissolution found empirically to favor the ratio toward complex formation were then analyzed to favor the formation of mixtures of inclusion complexes and non-inclusion complexes of cladribine and cyclodextrin in various proportions according to the invention.

Thus, for example, starting with more diluted cyclodextrin (ie, larger volumes of water than the volume of water used for the graphical solubility analysis) would logically provide more cladribine in solution, isolating more cladribine from complexation. During evaporation, some amount of solubilized cladribine will try to bind to the cyclodextrin in the non-inclusion complex. By changing the initial dilution, the specialist can shift the equilibrium towards the formation of an inclusion complex or a non-inclusion complex. Similarly, by increasing the complexation temperature, the solubility of cladribine in water can be increased, while the stability of inclusion complexes is reduced, thus 7/0 stimulating the formation of non-inclusion complexes. Thus, by changing the temperature of complex formation, the specialist can shift the equilibrium in the direction of the formation of an inclusion complex and a non-inclusion complex.

Finally, the complexation time can be varied to facilitate the formation of mixtures of inclusion complexes and non-inclusion complexes of cladribine and cyclodextrin according to the invention.

As shown below, it is possible to maximize cladribine in solid amorphous mixtures by introducing /5 additional cladribine into the solution (using more dilute cyclodextrin solutions, higher temperatures, and longer complexation times as shown above). When the solution is cooled, the highly amorphous nature of these cyclodextrins does not allow the passage of crystallization of an excess amount of cladribine, more than that which forms an inclusion complex with cyclodextrin; and upon freeze drying/lyophilization, the skilled person obtains an amorphous mixture of cladribine-cyclodextrin inclusion complex (which is amorphous) and amorphous free cladribine weakly bound to uncomplexed cyclodextrin (and even to complexed cyclodextrin) through the formation of hydrogen bonds, i.e. a complex not included

As shown in the Examples, this can be accomplished by maximizing solubility by raising the temperature (eg, to a temperature of about 502 to 802C), and stirring for many hours (up to 24 hours) before freeze-drying. The resulting mass/mass ratio was approximately 1:14 s and 1:11. The apparent optimum weight/weight ratio under these exemplified conditions is greater than or equal to approximately a 1:14 cladribine:cyclodextrin ratio. If too large an excess of cladribine is added to the complexation medium, then crystallization of some amount of cladribine occurs, which in turn would lead to the formation of some amount of crystalline cladribine in the product; this unwanted excess cladribine is not in solution and is not hydrogen-bonded to amorphous cyclodextrin and lowers the mass ratio. Therefore, it is desirable to carefully control the amount of excess cladribine, which is more than that which will form an inclusion complex only up to the amount that will dissolve in the solution. The desired amorphous mixture of the amorphous inclusion complex and the amorphous free co-cladribine may be referred to as a "cladribine-cyclodextrin complex" that includes both the inclusion complex and the non-inclusion/H-bonded complex. The inclusion complex is a cladribine complex, -

335 Inserted into the hydrophobic cavity of selected amorphous cyclodextrin, while the co-inclusion/H-bonded complex is amorphous free cladribine weakly hydrogen-bonded to cyclodextrin. It is estimated that approximately two-thirds (60-70905) of cladribine will be in the non-inclusion complex and the remaining one-third (30-4095) will be in the inclusion complex when the product is prepared as described below (1790 HP DSO solution, with 45 -509С7 temperature of complex formation "for about 9 hours); by increasing the percentage of cyclodextrin used and/or influencing the c temperature, products can be easily obtained in which a much larger proportion of the amorphous mixture is in the c form of the inclusion complex. In the case of a typical amorphous cyclodextrin, hydroxypropyl-D-cyclodextrin"" (HPVSOO), a mass ratio of cladribine:cyclodextrin of about 1:10 to about 1:16 is acceptable under the conditions given; it is expected that the ratio should be the same for hydroxypropyl-y -cyclodextrin under the same conditions. The obtained materials are characterized by rapid dissolution of bo cladribine in an aqueous medium. - Freeze drying, also known as lyophilization, consists of three main stages: first a freezing stage, then a primary drying stage and, finally, a secondary drying stage. Example 2, and presented below, provides details of lyophilization as performed on the batches described herein. This 20 procedure can further be optimized following the principles (described by Hiaosip (Spagie) Tapd and Misnaye! 3.

River! in Ripagtaseciis! Kezeagsi, Volume 21, Mo2, Rebgsagu 2004, 191-200), incorporated herein by reference in

T" in its entirety and can be controlled.

The method described above requires amorphous cyclodextrins, instead of initially crystalline cyclodextrins that have relatively low water solubilities, such as o-, p-, or y-cyclodextrin, 2,6-dimethyl-pD-cyclodextrin 22 and the like, because these cyclodextrins allowed b crystallization of cladribine in an excess of the former

HF) inclusion complex and, therefore, would not lead to the desired amorphous mixture. This method would also not be useful if cladribine were highly hydrophobic/lipophilic, because then the drug would not have the inherent water solubility/H-binding capacity and would not be able to provide the unambiguous mixture obtained in this description. However, in fact, cladribine has a water solubility of 5 mg/mL at room temperature 60, thus a significant amount of the drug will be simply soluble in the aqueous phase, especially at temperatures greater than room temperature; also, as in the case of NRDSO, for example, some cladribine will be hydrogen bonded to the 2-hydroxypropyl and free OH groups of glucose in the cyclodextrin through the two hydroxy functional groups found in the deoxyadenosine moiety of cladribine.

Cyclodextrins within the scope of this invention are amorphous derivatives of natural cyclodextrins: o-, D-,. and γ-cyclodextrins in which one or more hydroxy groups are substituted, e.g.

groups: alkyl, hydroxyalkyl, carboxyalkyl, alkylcarbonyl, carboxyalkyloxyalkyl, alkylcarbonyloxyalkyl, hydroxycarbonylalkyl or hydroxy-(mono or polyalkyl)alkyl; and in which each alkyl or alkylene moiety preferably contains up to six carbon atoms. Although it is usually referred to as a single element, amorphous cyclodextrin is actually a mixture of many different components, as the substituent groups can be located on different hydroxyl groups of the cyclodextrin backbone. This in turn leads to the amorphous nature of these cyclodextrins, which are indeed well known. In addition, these cyclodextrins can be obtained in variable degrees of substitution, for example, from 1 to 14, preferably from 4 to 7; degree of substitution represents the approximate average number of substituent groups on a cyclodextrin molecule, for example, the approximate number of 7/0 phydroxypropyl groups in the case of a hydroxypropyl-p-cyclodextrin molecule, and all such variations are within the scope of this invention. Substituted cyclodextrins that can be used in the present invention include polyesters, for example (as described in US Patent No. 3,459,731). Additional examples of substituted cyclodextrins include ethers in which the hydrogen atom of one or more cyclodextrin hydroxy groups is substituted with groups: C.alkyl, hydroxy-C.i.alkyl, carboxy-C.i.alkyl or C). valkyloxycarbonyl-C. and valkyl or their mixed 7/5 ethers. In particular, such substituted cyclodextrins are ethers in which the hydrogen atom of one or more cyclodextrin hydroxy groups is substituted by groups: C.isoalkyl, hydroxy-Solalkyl or carboxy-C.isoalkyl or even better by groups: methyl, ethyl, hydroxyethyl, hydroxypropyl , hydroxybutyl, carboxymethyl or carboxyethyl. The term "C. alkyl" should be understood as a group that includes linear and branched saturated hydrocarbon radicals containing from 1 to b carbon atoms, such as methyl, ethyl, 1-methylethyl, 1,1-dimethylethyl, propyl, 2-methylpropyl , butyl, pentyl, hexyl and the like. Other cyclodextrins believed to be useful for the same purpose include glucosyl-p-cyclodextrin and maltosyl-p-cyclodextrin. Particularly useful in the present invention are arbitrarily methylated p-cyclodextrin and polyesters such as hydroxypropyl-D-cyclodextrin, hydroxyethyl-pD-cyclodextrin, hydroxypropyl-y-cyclodextrin and hydroxyethyl-y-cyclodextrin, as well as sulfobutyl ethers, especially p -cyclodextrin sulfobutyl simple ether. In addition to simple cyclodextrins, branched cyclodextrins and cyclodextrin polymers can also be used. Other cyclodextrins are described,

And for example, in Spetisai! apa Riagtaseciisa! VyPape 28: 1552-1558 (1980); Makiduo deyo Mo. 6452 (28 Magsp 1983); Apdem Spent Yippee Ed. Epai 19: 344-362 (1980); US Patent Nos. 3,459,731 and 4,535,152; European

MoMo patents ER 0 149 197A and ER 0 197 571 A; PCT International Patent Application MOUUO90/12035; and Patent "

Applications of Great Britain JI 2,189,245). at

References describing cyclodextrins for use in the compositions of the present invention and providing methods for the preparation, purification, and analysis of cyclodextrins include the following: (Susiodehyp and)

Tesppoyudu ogvei Zgeyi, Kishmeg Asadetis Rybiivpegz (1988) in the section Susiodehiip5 ip REaptasetschisa!v; (p

Susiodehihip Spetiviu M. Her. Vepdeg and others, Zrgipdeg-Mepad, Vegip (1978); Aidmapsez ip Sagropuaga (e

From SPpetivigh, volume 12, under the editorship of M. Y. MMoyot, Asadetis Rgezv, Mem/ Mogk in the section "Te Zspagaipdeg Oehigipv" with

Oehieg Egepsp, pp. 189-260; Susiodehigipv apa (Peig Ipsiviop Sotriehez U. 527edi, AKadetiai Kiado,

Vydarev, Nipdagu (1982); I. Tarivzpi, Ase. Spent Kezeagsp, 1982, 15, pp. 66-72; MU. Zapdeg, Apdezhmapaiye

Spetiye, 92, pp. 343-361 (1981); A.R. Stoi et al., Tei(apedgop, 39, pp. 1417-1474 (1983); Igie et al., "

Rpagtaseciisa| Kezeagsi, 5, pp. 713-716 (1988); Rita and others, Ipi. 9. Ripagt. 29, 73 (1986); US Patent MoMo 4,659,696 and 4,383,992; German patents MoMo OE 3,118,218 and OE-3,317,064; and European Patent Mo ER 0 Z p 094 157A. Patents describing a hydroxyalkylated derivative of p- and y-cyclodextrin include Ria U.S. Pat. Nos. 4,596,795 and 4,727,064, MyPeg U.S. Pat.

Amorphous cyclodextrins for complexation with cladribine that deserve more attention include: hydroxyalkyl, for example, hydroxyethyl or hydroxypropyl, D- and y-cyclodextrin derivatives; co carboxyalkyl, for example, carboxymethyl or carboxyethyl, p- or y-cyclodextrin derivatives; p-cyclodextrin - sulfobutyl ether; and arbitrarily methylated p-cyclodextrin. 2-Hydroxypropyl-p-cyclodextrin c (HPVSO), 2-hydroxypropyl-y-dcyclodextrin (HRUSO), arbitrarily methylated p-cyclodextrin, p-cyclodextrin sulfobutyl ether, carboxymethyl-pD-cyclodextrin (SMVSO) and carboxymethyl-y-cyclodextrin o (SMUSO) are cyclodextrins of particular importance, especially hydroxypropyl-D-cyclodextrin and hydroxypropyl-y-cyclodextrin.

Compositions of an amorphous mixture of amorphous free cladribine and an amorphous, preferably saturated, cladribine-cyclodextrin inclusion complex for use in this invention can be obtained

Under conditions conducive to the formation of a complex in a liquid medium, as described and illustrated herein.

The resulting liquid compositions can then be converted into a dry form suitable for use in the form

F) solid oral or transmucosal dosage forms. One of ordinary skill in the art will appreciate that many approaches are available in the art to produce compositions as set forth herein. One available method, exemplified herein, includes the steps of mixing cladribine in an aqueous amorphous cyclodextrin solution, separating uncomplexed cladribine (eg, by filtration or centrifugation), and lyophilizing or freeze-drying the saturated solution to form a solid amorphous mixture.

Pharmaceutical compositions according to this invention may optionally include one or more excipients or other pharmaceutically inert components. One of the advantages of the present invention, however, is that the dosage forms of cladribine as provided herein can be prepared with the minimum amount of excipients necessary for molding and manufacturing in a particular form, such as a tablet or patch.

Excipients may be selected from those that do not interfere with cladribine, cyclodextrin, or complex formation.

Dosage forms are optimally prescribed in a pharmaceutically acceptable excipient with any well

Known pharmaceutically acceptable carriers, diluents, binders, lubricants, disintegrants, absorbents, flavorings, dyes and fillers.

Rpagtaseciisa! Ekhsiriepiv, MagseI YuOekKKeg Ips., Memzh/ MogkK and Vaze! (1998); And asptap and others., ed., Te Tneogu apa Rgasiise oi Shshproba Riaptasu, 372 ed., (1986); Pereptap et al., ed., Rpagtaseciisa! Oozade Rogtv,

Mags! Oekkeg Ips., Mem/ Hogk and Vazei! (1989); and Tne Naparsok oi Riagtaseciisa! Ekhsiriepiv, 372 ed., Ategisap 70 RIagtaseciisa| Avzzosiaiop and RIpagtaseciisa! Rgezv, 2000); see also Ketipdiopye Ripagtaseciisa| 5th ed., 18th ed., Seppago, Musk Rybiizpipd So., Eavsiop, RA (1990) and Ketipdiop: Te Zsiepse apa Rgasiise oi

Rpagtasu, IGirripsoi, UMiShate and UMiKipe, (19953). A simple solid oral dosage form consists of an amorphous mixture of amorphous free cladribine and an amorphous cladribine-cyclodextrin complex (predominantly saturated) as described above, i.e. a complex cladribine-dcyclodextrin complex 75 compressed with a small amount (e.g., about 195 by weight) of an acceptable binder substance or lubricant, such as magnesium stearate.

In certain cases, oral absorption can be further facilitated by the addition of various fillers and excipients to increase solubility or to increase penetration, for example, by modifying the microenvironment.

The methods and pharmaceutical compositions disclosed herein offer novel therapeutic methods for the treatment of patients in need of treatment with cladribine. As shown in this description, the present invention addresses the problems of low bioavailability traditionally associated with the oral use of cladribine.

The compositions of this invention are particularly suitable as methods of treatment of any cladribine-sensitive c

Diseases Some disease states responsive to cladribine are well described in the literature (see below). For any target disease condition, an effective amount of a complex cladribine-cyclodextrin complex, i.e. an amorphous mixture of optimized amorphous saturated cladribine - amorphous cyclodextrin complex with amorphous free cladribine, as described above (eg, an effective amount for the treatment of multiple sclerosis, rheumatoid arthritis or leukemia). "

The term "therapeutically effective amount" or "effective amount" is used to refer to treatment in doses effective to achieve the desired therapeutic effect. Therapeutically effective doses reported in the literature include doses for hairy cell leukemia (0.09 mg/kg/day for 7. :CO days), for multiple sclerosis (from about 0.04 to about 1.Omg/kg/day |see US Patent Mo5,506,214); "- for other diseases, see also US Patent MoMo5,106,837 (autohemolytic anemia); 5,310,732 (inflammatory

Intestinal disease); 5,401,724 (rheumatoid arthritis); 5,424,296 (malignant astrocytoma); 5,510,336 so (histiocytosis); 5,401,724 (chronic myeloma leukemia); and 6,239,118 (atherosclerosis))|.

In addition, various dosage amounts and dosage regimens are described in the literature for use in the treatment of multiple sclerosis; see, for example: Kotipe and others, Rgoseedipdz oi (pe Avzzosiaiop oi Ategisap

Rpuzisiape5, volume 111, Moi, 35-44 (1999); Beru and others, Te Sapadiap docsgpa! oi MeyigoYodisaIi Zsiepsev, 25, 295-299 (1998); TopogeMa and others, Sitepi Oripiop ip Ipmeviidaiopa! Ogydv, 2 (12), 1751-1756 (2001); Kise Zs and others, Meyigoiodu, 54, 1145-1155 (2000); and Kaghizzop and others, Vgyivpy doigpa! et al., 116, 538-548 (2002); all of which are incorporated herein by reference in their entirety and may be incorporated herein by reference. attention While these formulations optimize the bioavailability of cladribine after oral administration, it should be noted that even optimal bioavailability from oral dosage forms is not expected to reach the bioavailability obtained after intravenous administration, especially at initial time points. Therefore, it is often acceptable to increase the dose proposed for intravenous administration to achieve an acceptable dose for administration in a solid oral dosage form. At this time, it is envisaged that for the treatment of multiple sclerosis 1Omg of cladribine in a soluble cladribine-cyclodextrin complex of 250 complex in a soluble solid dosage form could be used once a day for five to seven days in the first month, and then repeated during another period of five to seven days in the second month, that trip would be followed by ten months without treatment. Alternatively, the patient could use 10 mg of cladribine in a soluble complex cladribine-cyclodextrin complex in soluble dosage form once a day for five to seven days per month for six months, followed by eighteen months without treatment. For additional information regarding dosage, |see also Previous

GF! US Patent Application Mo033935-011, and US Provisional Patent Application Mo033935-0121), both entitled "SiIadgiripe Keditep bog Tgeaipo Miyyrie Zsiegoziv", both filed March 25, 2004 and incorporated herein by reference in their entirety.

Additionally, the skilled artisan should appreciate that the therapeutically effective amount 60 of cladribine, the use of which is described herein, can be reduced or increased by careful selection and/or use of cladribine according to the present invention with another active ingredient. Therefore, the present invention provides a method of tailoring application/treatment to certain extreme needs specific to each individual mammal. Therapeutically effective amounts can be easily determined, for example, empirically, starting with relatively low amounts and gradually increasing the amount of the agent while simultaneously evaluating the positive effect.

As indicated in the previous section, the use of cladribine in accordance with the present invention may be accompanied by the use of one or more additional active ingredients for the treatment of a cladribine-sensitive condition. The additional active ingredient will be used by administration and at dosage amounts and frequencies appropriate for each additional active ingredient and the condition being treated. For example, in the treatment of multiple sclerosis, other useful drugs include interferon-beta (Keri, Vefazegope/Weifagegope?, AmopexF), which is identical to a protein of natural origin found in the human body; glatiramer acetate (Sorahope?F), a random chain (polymer) of glutamic acid, lysine, alanine and tyrosine amino acids; natalizumab (Apiedhep?F), a monoclonal antibody; alemtuzumab (Satraii-1NF)), guianized anti-CO52 monoclonal antibody; 4-aminopyridine (also known as 7/0 8-AP and Fampridine), a drug that blocks potassium channels in neurons; and amantadine, an antiviral agent that improves muscle control and reduces muscle stiffness and is used to relieve fatigue symptoms in multiple sclerosis, a goal for which pemoline (Sueg) and

Y-Carnitine (product of natural origin). In the treatment of hairy cell leukemia, additional active ingredients may include interferon-alpha, pentostatin, fludarabine, rituximab (anti-CO 20 monoclonal /5 antibody) and anti-CO 22 recombinant immunotoxin VI 22; other additional active ingredients may be evaluated in other types of leukemia. When treating rheumatoid arthritis, there are many other active ingredients that can be chosen. These include NSAIDs (non-steroidal anti-inflammatory drugs), which are of three types: salicylates, such as aspirin, traditional NSAIDs, such as ibuprofen and indomethacin, and COX-2 inhibitors, such as celecoxib (CiIergexF), rofecoxib (MioxxF), meloxicam (Mobis?), valdecoxib (Vehigaf), lumiracoxib (Rgehide?F) and etoricoxib (AgsohiafF). Other drugs useful in the treatment of rheumatoid arthritis that can be used in connection with the present invention include OMAKOZ5, glucocorticoids, biological response modifiers and non-M5AIU analgesics. OMACOZ are disease-modifying antirheumatic drugs that include methotrexate, plaquenil, leflunomide (Agamaf), salazosulfapyridine, gold, penicillamide, cyclosporine, methylcyclofosamide, and imuran. Glucocorticoids include dexamethasone, prednisolone, triamcinolone and many others. Biological response modifiers (which restore the immune system's ability to fight disease) include etanercept (EpgeF), a tumor necrosis factor (TNF) inhibitor, infliximab (KetisadeFf), which is also an anti-TME drug, anakinra (KipegeF), a selective II/-1 blocker, and NitigaF, a human monoclonal antibody, which is another anti-TME drug. Non-NSAID analgesics include acetaminophen, as well as narcotic analgesics such as dihydroxycodeinone, oxycodone, and propoxyphene. Generally speaking, those drugs that work by a mechanism different from that of cladribine are extremely useful for adjunctive therapy with the cladribine composition presented herein. Those drugs that are effective for the oral route of administration and that are compatible with soluble cladribine complexes in a single dosage form can be included in soluble dosage forms; otherwise, of course, they should be applied separately in quantities, with frequency and routes of application acceptable to them. co

As used herein, "treating" means reducing, preventing, inhibiting, controlling, ameliorating, and/or reversing the symptoms of a patient treated with a compound of the present invention as compared to the symptoms of a patient not treated in accordance with the present invention. The practitioner should take into account that the complexes, compositions, dosage forms and methods presented in this description "must be used in conjunction with ongoing clinical studies conducted by a qualified specialist (therapist or veterinarian) in this field to determine the next therapy . Such a study

And will help and inform the assessment of whether to increase, decrease or continue to use y?” a certain therapeutic dose, and/or change the method of application.

The methods of this invention are intended for use by any subject/patient who may experience the positive effects of the methods of this invention. Thus, in accordance with the present invention,

Go! the terms "subjects" as well as "patients" include humans as well as non-human subjects, including domesticated animals. - Any acceptable materials and/or methods known to specialists qualified in this field may 2) be used in the implementation of this invention. However, better materials and methods are described. Materials, reagents, and the like referred to in the following description and examples may be obtained from commercial sources unless otherwise noted. The following examples are intended to further illustrate certain preferred embodiments of the present invention and are not intended to limit it. Those skilled in the art will know or be able to establish, using no more than routine experimentation, acceptable equivalents to certain substances and procedures disclosed herein.

Examples (Ф) Example 1 of the study of phase solubility ka The study of phase solubility was carried out as follows.

An excess of cladribine was added to cyclodextrin solutions of various concentrations: hydroxypropyl-D-cyclodextrin (HPVSO) and the complex was treated as described in Example 2 below. Excess, undissolved cladribine was removed by filtration. The amount of cladribine in the complexation solution was measured to obtain one of the data processing results (points on the curve). This process was repeated with various known concentrations of cyclodextrin until several curve points were obtained. These curve points were then plotted, with each of these points representing the amount of cladribine 65 that could be dissolved in water with a certain concentration of cyclodextrin. The points on the line constructed from the data processing results represent the ratio for the product. One of ordinary skill in the art should appreciate that similar results will be obtained if an excess of cyclodextrin is added to solutions of cladribine of known concentration.

A graph of the dependence of the obtained molar concentrations of cladribine on cyclodextrin is constructed and presented graphically. A typical phase solubility diagram is shown in Fig. The plotted lines for the cladribine-NRVSO complex represent the solubility of cladribine for the test conditions, that is, the ratio of the concentration of cladribine to the concentration of cyclodextrin. The area above each of the plotted lines represents the conditions under which excess undissolved cladribine is present. The area below each of the plotted lines represents the conditions under which cyclodextrin is in excess. 70 The dependence graph for cladribine-HP DSO, shown in Fig., is practically linear; this shows the formation of a 1:1 complex in which one drug molecule forms a complex with one cyclodextrin molecule. On

The figure also shows that additional cyclodextrin is required to hold cladribine in the complex.

For example, approximately 0.14 mol of НРДСО is necessary to hold approximately 0.049 mol of cladribine dissolved under the selected conditions, which will ultimately provide the formation of an amorphous mixture of an amorphous, 75 mostly saturated, cladribine-НРДСО inclusion complex and amorphous free cladribine (in the form of a non-inclusion complex). . Under the conditions of Example 2 below, a significant portion of the cladribine in the product would be expected not to be in an inclusion complex, but rather in an amorphous form that is weakly held in a homogeneous mixture, in addition, by hydrogen bonding as a non-inclusion complex .

Example 2

Preparation of cladribine-cyclodextrin complex for human trials

Cladribin forms a complex with NRDSO in the following way.

In 825 ml of distilled water, 172.5 g of hydroxypropyl-p-cyclodextrin are dissolved (forming about 1790 solution), then cladribine is added and the mixture is stirred at a temperature of about 45 to about 502C for about nine hours. Stirring is continued for an additional 6-9 hours at room temperature. Any undissolved cladribine is removed by filtration and the solution is cooled to room temperature. To form an amorphous mixture of amorphous cladribine-cyclodextrin complex and amorphous free cladribine, the aqueous cladribine-cyclodextrin solution is dried by lyophilization before administration into solid oral tablets. The lyophilization procedure includes the step of freezing by rapidly bringing the complexation solution to a temperature of from about -40 °C to about -80 °C (eg, about -45 °C) for a period of time from about 2 to 4 hours (preferably from about 3 to 4 hours), followed by a primary drying stage at about -252C for about 80-90 hours, usually at low pressure, and then a secondary drying stage at about 302C for about 15-20 hours. -

The product obtained by the above basic procedure can be analyzed using ! ! ! ! d)

HRIS (using a Nuregzvi O05 3-micron column and an acetonitrile-based mobile phase, with UV detection at 26b4nm) to determine the mass ratio of cladribine to cyclodextrin in the final product. The compositions of the final products can be further characterized by methods known in the art, including, for example, character inspection, determination of total impurity content using NRIS, determination of water content using a Kagi Rizspeg titrator, determination of the solubility profile using a standard method, for example, using О5Р"711"Arragaiyiz II device and

UV detection at 26b4nm, establishing the homogeneity of the content and performing a quantitative study with the help of NRI C analysis of the active ingredient.

Two batches of the cladribine/cyclodextrin product, EOO4 and EBO5, were prepared by the procedure described above, as shown below: c Purified water (825 mL) was heated at 482C (target range 4592 to 5022) in a 1 liter glass flask by immersion in a water bath. The heated water was stirred to achieve a controlled central vortex. - 2-hydroxypropyl-D-cyclodextrin (172.50g) was weighed and slowly added to heated water over 40 minutes. c The resulting solution was stirred for an additional 10 minutes to ensure complete dissolution of the Cyclodextrin. Cladribine (12.00g for E0BO4 and 18.75g for EBOB) was weighed and added to a stirred solution of cyclodextrin, which turned cloudy before becoming clear. The resulting clear solution was kept at 482C and constantly stirred for 9 hours. Stirring was continued for another 7 hours until the solution cooled to room temperature.

The use of large amounts of cladribine in the preparation of EOO5 was part of an attempt to optimize this procedure; however, it was found that the initial amount of cladribine in that case was too much and precipitation was observed at the end of the cooling stage for batch EBO5. The solution was filtered to remove

F) sediment. The analysis of the obtained product revealed (content of the component according to the analysis results - 87.2965) that 16.35 g of cladribine is included in the cyclodextrin complex in the case of EOOB5. Batch EO0O4 did not require any filtering, showing that the quantities used in the preparation of EO04 were more acceptable and that EBO5 because the procedure could have been optimized by starting with a smaller amount of cladribine (16.35g instead of 18.75g), thus avoiding filtering stage.

After cooling to room temperature and, in the case of EBOB5, filtering, the solutions were poured into 100ml lyophilization tubes (2Oml of solution in each tube), the filled tubes were partially stoppered and lyophilized. Lyophilization included freezing at -452C for approximately 200 minutes, a primary drying phase at -252C at 100mTorr for approximately 5200 minutes, and a secondary drying phase at 302 for approximately 1080 minutes as follows:

zo anayut 00006000 and 8 Pera vyuushuvanya| bo 00000005 (8 Secondary drying of ZUS 50001000 in Zhnschevasvrova 0 of 0 Probrzysayutuvayuuyu 12 E0O04 and ROB batches of the cladribine/cyclodextrin product obtained according to the procedure described above were analyzed with the help of NRI C (using a Norwegian O05 3-micron column and a mobile phase based on acetonitrile, with UV detection at 26b4nm) and empirically determined that they have the following characteristics:

These products were analyzed by Y5C thermograms and X-ray diffraction to determine any free crystalline cladribine in the lyophilized material. Importantly, the samples did not show any transition in the 21020-2302С region, which is associated with the melting of crystalline cladribine. In both cases, no significant thermal activity was recorded in the 21020-2302C region, suggesting that the complexes obtained at the end of lyophilization do not have any significant amount of free crystalline cladribine, taking into account the sensitivity of the analytical method (up to 395 wt/wt). This conclusion is confirmed by the absence of peaks for C crystalline cladribine on the scan of the X-ray diffraction method for both EOO4 and O complexes

E0Oo5.

These products are amorphous mixtures of amorphous cladribine-"HP DVSO complex of inclusion and i. of amorphous free cladribine, attached by hydrogen bonds to cyclodextrin in the form of a "- non-inclusion" complex. The resulting mass ratios of cladribine:HRDVSO were approximately 1:14 and 1:11.

Generally speaking, the amorphous mixtures within the scope of this invention have mass ratios of co-cladribine:HPrSOB from about 1:10 to 1:16.

Example C

Preparation of Oral Tablets Tablets were produced using batches of amorphous mixtures of EOO4 and RBOB5 described in Example 2 for use in a clinical trial. Batch MO120 was produced using a mixture of cladribine-2-NRDSO of the OBROB complex with a batch size of 3,000 tablets and batch MO126 was produced using a mixture of cladribine-NRDSO of the EOO4 complex with a batch size of 800 tablets. Control compositions for two lots are shown in Table IP. Lot MO120 was a 3.0mg tablet and Lot MO126 was a 10mg tablet for the clinical trial. sa - Component Batch number o 00 dev pattyutmnya

Mixing and breaking rum ROB 00000000 7 Mixing and breaking rum set Р00Э 00000000 shaft

Vvyuyu 000000000001000010yu1yuyu: "Equivalent to 3.0mg of cladribine per tablet

GF) "x Equivalent to 10.Omg cladribine per tablet o The following table describes the method of production of Batch MO120 and MO126 tablets. in a 40-mesh sieve. where 5. | Mixing the final mixture for 5 minutes at 12 rpm.

The tablets of both Lot MO120 3.Omg tablets and Lot MO126 10.Omg tablets were round, flat on one side with a beveled edge and slightly convex on the other side. A batch of MO120 Z,Omg tablets had an average weight of 100 mg, a thickness of 2.7 mm, a friability of 0.295, a hardness of 4 AKr, and a dissolution time of 3 minutes. A batch of MO126 10.0mg tablets had an average mass of 198mg, a thickness of 4.2mm, a friability of 195, a hardness of 2.8Cr and a disintegration time of 5 minutes and 42 seconds.

Lot MO120 3.Omg tablets and Lot MO126 10.Omg tablets were used in the clinical trial summarized in Example 5 below. then Example 4

Clinical trial: relative bioavailability

The purpose of this study is to evaluate the relative bioavailability of three oral formulations of cladribine: (1) a cyclodextrin-based formulation according to the present invention (Tablet 1: EOBO5 complex, i.e.

Batch MoMO120 tablets described above); (2) mucoadhesive composition (Tablet 2: contains 3.0mg of cladribine, then 10mg of Sagroroi 710 IU, 22.2mg of dicalcium phosphate, 64.Zmg of lactose and 0.5mg of magnesium stearate, Batch Mo MO121); and (3) hard gel capsule (Capsule containing 3.0mg cladribine, 5.00mg Sagroroa 974R, 91.33mg Amise! RNIO1, 100.Omg AmiseI RNIO2, 0.2mg colloidal silicon dioxide and 0.5mg magnesium stearate, Batch MoKk0OZ3030) compared with one specific subcutaneous application of cladribine (control composition) in patients with M5 (multiple sclerosis). 720 This trial was a 2-center, open-label, randomized, 4-channel crossover single-dose trial using twelve patients with M5. Patients were randomized to receive three different defined oral doses (Z,Omg) and a specific subcutaneous dose of Z,Omg. The four-day treatment was separated by drug-free intervals of at least 5 days. In each treatment period, blood samples were collected over a 24-hour period to assess the amount of cladribine in plasma. cm y y y y - ;

The concentration of cladribine in plasma was measured using the NRI C/M5/M5 method. Using this method, (o) the relationship between concentration ratios and peak area ratios was found to be linear in the range 10Opg/ml to 50.00Opg/ml for cladribine. The limit of quantification was 1OOpg/ml. Sample analysis was performed in 16 batches. No calibrator was excluded from the selection of the calibration curve by points, and the accuracy of each quality control sample met CI R requirements.

576 clinical plasma samples were analyzed and cladribine concentration values were determined. The results (a) were combined and summarized in the tables presented below (Tables M and MI). In these tables, the following definitions are acceptable: T is the time to reach the maximum concentration in plasma; T 4/»o is the half-life of cladribine in plasma; The peak represents the maximum concentration of cladribine in plasma; -

AMC; is the area under the curve for measured data from zero extrapolated to infinity; co

АСМсС, is the area under the curve for the measured data (from zero to the last time point); Ser. geom. is the geometric mean; SM is the deviation coefficient (relative standard deviation); And I. represents the lower limit; AI. is an upper limit. "

And 7 s for the study of cladribine, obtained with the help of non-separative analyses, (p-12).

Iz» Pharmacokinetic Z,Omg subcutaneously Zmg Tablet 1 Zmg Tablet 2 Zmg Capsule parameter rod et t so geom. 80 geom. | 80 geom. I8O0 geom. 1850 (o) (o) (o) (o) -

F

» z l (F) Table MI

Kk The ratio of the pharmacokinetic parameters of oral administration to the pharmacokinetic parameters of subcutaneous administration and the corresponding two-sided 9095 confidence intervals for the cladribine test (p-12) 6o Pharmacokinetic Z,Omg Tablet 1 Zmg Tablet 2 Zmg Capsule parameter

Ratio" OS Ratio" OS Ratio" OS

A!ySide 43.1 ZB, Sv 31.8, 38.9 321, 2 46A 47.0 65 dis, 41.9 34.6, 372 30.7, 37.6 31.0, 50.8 45, 0 45.5

"Ratios (dose-normalized) and corresponding 9595 G, obtained through the inverse transformation of IOSD-transformed data.

Example 5

Clinical trial: dose response and absolute bioavailability

The objective of this trial was to evaluate the systemic availability of cladribine after oral administration of two different fixed oral doses, compared with fixed intravenous administration (control formulation) in patients with M5 (multiple sclerosis), and to evaluate the safety and tolerability of cladribine in this population . 70 This trial was a single-center, open-label, randomized, three-channel crossover single-dose trial using twenty-six patients with M5. Patients were randomized to three different defined oral doses (3,Omg and 10,Omg) and a specific subcutaneous dose of 3,Omg (administered as a 1-hour infusion). The three-day treatment was separated by drug-free intervals of at least 5 days. In each treatment period, blood samples were collected over a 24-hour period to assess plasma cladribine.

The concentration of cladribine in plasma was measured using the NRI C/M5/M5 method. Using this method, the ratio of concentration to peak area ratios was found to be linear over the range of 100 pg/ml to 50.000 pg/ml for cladribine. The limit of quantification was 1OOpg/ml.

Sample analysis was performed in 16 batches. Except for the first batch (which had to be discarded due to an instrument error), all other batches could be used. No calibrator was excluded from the fitting of the calibration curve by points, and the accuracy of each quality control sample met CI R requirements.

858 clinical plasma samples were analyzed and cladribine concentration values were determined. The results were combined and summarized in the tables presented below (Tables MII-X). In these tables, the following definitions are acceptable: T pah represents the time to reach the maximum concentration in plasma; T 4/ is the c half-period of cladribine in plasma; Cmax is the maximum plasma concentration of cladribine; AOS; (U represents the area under the curve for measured data from zero extrapolating to infinity; AOS ; represents the area under the curve for measured data (from zero to the last time point); Geom. Mean is the geometric mean; CM is the coefficient deviation (relative standard deviation); HI. is the lower limit; І. is the upper limit; c is the average variability; осв" is the average C variability between subjects; сш is the average variability within subjects; CMT is the overall coefficient of variation, and SM is the coefficient of variation within the subjects. c - » z for the study of cladribine, obtained with the help of non-separative analysis, (n-26)

NYM nni vi nn zi vni nis KI" (o) (o) (o) "o shchi s:" with her 01111 nokoyudntsht toloki "su - zr/average for T pakh T95 and CM9U5 geometric mean for Stakh LYOSIipT As. -

Mn Table MI (ав) 50 The ratio of pharmacokinetic parameters of oral administration to pharmacokinetic parameters of intravenous administration and the corresponding lower limit (HH) iz" for a one-sided 9595 confidence interval for the cladribine test p-26).

Pharmacokinetic parameter Oral use 3.0 mg 10.0 mg

Ratio" her Ratio" her.

Gee! A!ysSipe 34.5 31.7 39.1 35.9 dis, 34.0 31.2 39.4 361 ime) "Ratios (dose-normalized) and the corresponding 9595 GIs obtained through the inverse transformation of Iyusd-transformed data 60 вв 0 спдяня 00100010

Stakh 112.6 951 133.3

"Ratios (dose-normalized) and corresponding 9095 GIs obtained through the inverse transformation of IOSD-transformed data

Table X

Components of deviation for the cladribine test (p-26)

Between subjects (St?) 0.оз8о0 0.0487 0.0492

Within the subject (US?) 0.1315 0.0330 0.0357 and styu sum 0)

Where pharmacokinetic parameters are dose-regulated and SU- uvhrye23-1.

The above should be considered as an illustration of only the basic principles of this invention. Furthermore, it will be apparent to one skilled in the art that various modifications and changes may be made to the claimed invention without limiting the present invention to the particular interpretations and developments shown and described, and accordingly, all reasonable modifications may be addressed. and their equivalents falling within the scope of the claimed invention. with

Claims (75)

The formula of the invention of 1. A pharmaceutical composition including a complex cladribine-dcyclodextrin complex, which is a homogeneous amorphous mixture of (a) an amorphous inclusion complex of cladribine with amorphous cyclodextrin "Ezo" and (B) amorphous free cladribine bound to amorphous cyclodextrin as a non-inclusion complex, prescribed in a solid oral dosage form. at 2. Pharmaceutical composition according to claim 1, which differs in that the complex is saturated with cladribine. co 3. The composition according to claim 1 or 2, which is characterized by the fact that the amorphous cyclodextrin is hydroxypropyl-cyclodextrin, hydroxypropyl-β-cyclodextrin, arbitrarily methylated p-cyclodextrin, (7-carboxymethyl-p-cyclodextrin or sulfobutyl-p-cyclodextrin. (uh) 4. The composition according to claim 1 or 2, which differs in that the amorphous cyclodextrin is hydroxypropyl-β-cyclodextrin. 5. The composition according to claim 1 or 2, which differs in that the amorphous cyclodextrin is hydroxypropyl-""Y-cyclodextrin. - with 6. The composition according to any one of claims 1-3, which is characterized in that the mass ratio of cladribine and "» amorphous cyclodextrin is from about 1:10 to about 1:16. " 7. The composition according to item b, which differs in that the amorphous cyclodextrin is 415 hydroxypropyl-pR-cyclodextrin. Ge | 8. The composition according to claim 7, which differs in that the mass ratio of cladribine and - hydroxypropyl-p-cyclodextrin is approximately 1:14. at 9. The composition according to claim 7, which differs in that the mass ratio of cladribine and about 50 hydroxypropyl-P-cyclodextrin is approximately 1:11. 10. The composition according to item b, which differs in that the amorphous cyclodextrin is c». I hydroxypropyl- "-cyclodextrin. 11. The composition according to any of claims 1-10, which is characterized by the fact that the approximate molar ratio of 5B cladribine and amorphous cyclodextrin corresponds to the point located on the phase solubility diagram for saturated complexes of cladribine in variable concentrations of cyclodextrin. (F; 12. The composition of any one of claims 1-11, wherein from about 30 to about 40 percent by weight of the cladribine is in the inclusion complex (a) and from about 70 to about 60 percent by weight of the cladribine is in the complex non-inclusion (b). 60 13. The use of a complex cladribine-cyclodextrin complex, which is a homogeneous amorphous mixture of (a) an amorphous inclusion complex of cladribine with amorphous cyclodextrin and (b) amorphous free cladribine bound to amorphous cyclodextrin as a non-inclusion complex, in the composition of a solid oral dosage form, for use in the treatment of symptoms of a cladribine-sensitive condition. 14. Application according to claim 13, which is characterized by the fact that the complex is saturated with cladribine. 65 15. Use according to claim 13 or 14, characterized in that the cladribine-sensitive condition is selected from the group consisting of multiple sclerosis, rheumatoid arthritis and leukemia. 16. Use according to claim 15, which differs in that the cladribine-sensitive condition is multiple sclerosis. 17. Application according to claim 13, 14, 15 or 16, which is characterized by the fact that the amorphous cyclodextrin is hydroxypropyl-p-diclodextrin, hydroxypropyl-7-diclodextrin, arbitrarily methylated p-cyclodextrin, carboxymethyl-p-cyclodextrin or sulfobutyl-p- cyclodextrin. 18. Use according to any one of claims 13-17, characterized in that the mass ratio of cladribine to amorphous cyclodextrin is from about 1:10 to about 1:16. 19. Use according to any of claims 13-18, which is characterized by the fact that the amorphous cyclodextrin is 70 hydroxypropyl-p-cyclodextrin. 20. Application according to claim 19, which differs in that the mass ratio of cladribine and hydroxypropyl-p-cyclodextrin is approximately 1:14. 21. Application according to claim 19, which differs in that the mass ratio of cladribine and 75 hydroxypropyl-P-cyclodextrin is approximately 1:11. 22. Use according to any one of claims 13-18, which is characterized by the fact that the amorphous cyclodextrin is hydroxypropyl-7-cyclodextrin. 23. The use of any one of claims 13-22, wherein from about 30 to about 40 percent by weight of the cladribine is in the inclusion complex (a) and from about 70 to about 60 percent by weight of the cladribine is in the non-inclusion complex (b). 24. The use of a complex cladribine-cyclodextrin complex, which is a homogeneous amorphous mixture of (a) an amorphous inclusion complex of cladribine with amorphous cyclodextrin and (b) amorphous free cladribine bound to amorphous cyclodextrin as a non-inclusion complex, in the composition of a solid oral dosage form , to increase the oral bioavailability of cladribine. 25. Application according to claim 24, which differs in that the complex is saturated with cladribine. Gee) 26. Application according to claim 24 or 25, which is characterized by the fact that the amorphous cyclodextrin is hydroxypropyl-p-diclodextrin, hydroxypropyl-7-diclodextrin, arbitrarily methylated p-cyclodextrin, carboxymethyl-p-cyclodextrin or sulfobutyl-p-cyclodextrin. "Her 27. Use according to any one of claims 24-26, characterized in that the mass ratio of cladribine to amorphous cyclodextrin is from about 1:10 to about 1:16. 28. Use according to any of claims 24-27, which is characterized by the fact that the amorphous cyclodextrin is me) hydroxypropyl-p-cyclodextrin. "- 29. Application according to claim 28, which differs in that the mass ratio of cladribine and co-hydroxypropyl-p-cyclodextrin is approximately 1:14. ZO. Application according to claim 28, which differs in that the mass ratio of cladribine and hydroxypropyl-p-cyclodextrin is approximately 1:11. " 31. Application according to any of claims 24-27, which is characterized by the fact that the amorphous cyclodextrin is (H(Sh with hydroxypropyl-7-cyclodextrin. "» 32. The use of any one of claims 24-31, wherein from about 30 to about 40 percent by weight of the cladribine is in the inclusion complex (a) and from about 70 to about 60 percent by weight of the cladribine is in the complex non-inclusion (b). 33. A complex cladribin-cyclodextrin complex, which is a homogeneous amorphous mixture (a) with an amorphous inclusion complex of cladribin with amorphous cyclodextrin and (B) amorphous free - cladribin bound to amorphous cyclodextrin as a non-inclusion complex. 34. The complex according to claim 33, saturated with cladribine. at 35. The complex according to claim 33 or 34, which differs in that the amorphous cyclodextrin is 20 hydroxypropyl-β-cyclodextrin, hydroxypropyl-β-diclodextrin, arbitrarily methylated p-cyclodextrin, T» carboxymethyl-p-cyclodextrin or sulfobutyl-p- cyclodextrin. 36. The complex according to claim 33 or 34, which is characterized by the fact that the amorphous cyclodextrin is hydroxypropyl-p-cyclodextrin. 37. The complex according to claim 33 or 34, which differs in that the amorphous cyclodextrin is HF) hydroxypropyl-7-cyclodextrin. ka 38. The complex according to any one of claims 33-35, characterized in that the mass ratio of cladribine to amorphous cyclodextrin is from about 1:10 to about 1:16. in 39. The complex according to claim 38, which differs in that the amorphous cyclodextrin is hydroxypropyl-p-cyclodextrin. 40. The complex according to claim 39, which differs in that the mass ratio of cladribine and hydroxypropyl-p-cyclodextrin is approximately 1:14. 65 41. The complex according to claim 39, which differs in that the mass ratio of cladribine and hydroxypropyl-p-cyclodextrin is approximately 1:11. 42. The complex according to claim 38, which is characterized by the fact that the amorphous cyclodextrin is hydroxypropyl-7-cyclodextrin. 43. The complex of any one of claims 33-42, wherein from about 30 to about 40 percent by weight of the cladribine is in the inclusion complex (a) and from about 70 to about 60 percent by weight of the cladribine is in the non-inclusion complex (b). 44. The method of obtaining a complex cladribine-cyclodextrin complex, which includes stages: 70 () combining cladribine and amorphous cyclodextrin in water at a temperature from about 40 to about 80 2С and maintaining the specified temperature during a period from about 6 to about 24 hours; (i) cooling the obtained aqueous solution to room temperature; and (iii) lyophilization of the cooled solution to obtain an amorphous product. 45. The method according to claim 44, which additionally includes a filtering stage after stage (ii). 75 46. The method according to claim 44 or 45, which is characterized by the fact that stage (i) is performed at a temperature from about 45 to about 60 es. 47. The method according to any one of claims 44-46, characterized in that the stage (ii) is performed at a temperature of from about 45 to about 50 20. 48. The method according to claim 46 or 47, which is characterized by the fact that the stage (ii) is performed during mixing. 49. The method of claim 48, wherein step (s) is performed over a period of time from about 6 to about 9 hours. 50. The method according to any one of claims 44-49, characterized in that the step(s) is performed over a period of time from about 6 to about 9 hours. 51. The method according to any one of claims 44-50, characterized in that step (ii) includes an initial freezing step in which the solution is cooled to a temperature of about -40 to about -80 °C and held at said temperature for from about 2 to about 4 hours. at 52. The method according to claim 51, which is characterized by the fact that at the primary stage of freezing stage (ii) the solution is cooled to a temperature of approximately -45 C. 53. The method according to any one of claims 44-52, which is characterized in that 12.00 parts by weight of cladribine and 172.50 parts by weight of hydroxypropyl-p-cyclodextrin are introduced in stage (i). at 54. The method according to any one of claims 44-52, which is characterized in that 16.35 parts by weight of cladribine and 172.50 parts by weight of hydroxypropyl-p-cyclodextrin are introduced in stage (i). at 55. The method according to claim 53 or 54, which differs in that 825 parts by volume of water are introduced into the stage (Her). -- 56. The method according to any of claims 44-55, which is characterized by the fact that the lyophilization stage (ii) includes: a) an initial stage of freezing, in which the complexation solution is brought to a temperature from about -40 °C to about -80 °C C for 2 to 4 hours; p) the primary stage of drying at a temperature of approximately -25 °C for approximately 80-90 hours; and c) a secondary drying stage at a temperature of about 30 °C for about 15-20 hours. " 57. The method according to claim 56, which differs in that the stage (a) of lyophilization is carried out at a temperature of approximately - with -459С2 for approximately 3-4 hours. and 58. The method according to claim 56 or 57, which differs in that stage (B) of lyophilization is carried out at a pressure of about 100 mTorr. 59. A pharmaceutical composition obtained by a method comprising the steps of: () combining cladribine and amorphous cyclodextrin in water at a temperature of about 40 to (ee) about 80 C and maintaining said temperature for a period of about 6 to about 24 hours ; - (iii) cooling the obtained aqueous solution to room temperature; (ii) lyophilization of the cooled solution to obtain an amorphous product; and o (m) formulating the amorphous product into a solid oral dosage form. at 20 60. Pharmaceutical composition according to claim 59, which is characterized by the fact that the method additionally includes a filtering stage after stage (i) or (iii). her" 61. Pharmaceutical composition according to claim 59 or 60, which is characterized by the fact that the stage (th) of the method is performed at a temperature from about 45 to about 60 es. 62. Pharmaceutical composition according to any one of claims 59-61, which is characterized in that stage (i) is performed at a temperature of about 45 to about 50 es. GF) 63. The pharmaceutical composition according to claim 61 or 62, which is characterized by the fact that the stage (s) of the method are performed during mixing. yes 64. The pharmaceutical composition according to claim 63, which is characterized in that the step(s) of the method is carried out over a period of time from about 6 to about 9 hours. 60 65. A pharmaceutical composition according to any one of claims 59-64, wherein the step(s) is performed over a time period of from about 6 to about 9 hours. 66. A pharmaceutical composition according to any one of claims 59-65, characterized in that step (iii) includes an initial freezing step, in which the solution is cooled to a temperature of from about -40 °C to about -80 °C and held at said temperature for about 2 to about 4 hours. bo 67. Pharmaceutical composition according to claim 66, which is characterized in that at the primary stage of freezing the stage (iii) the solution is cooled to a temperature of approximately -45 2С. 68. A pharmaceutical composition according to any one of claims 59-67, which is characterized in that 12.00 parts by weight of cladribine and 172.50 parts by weight of hydroxypropyl-p-cyclodextrin are introduced in stage (ii) of the method. 69. A pharmaceutical composition according to any one of claims 59-67, characterized in that 16.35 parts by weight of cladribine and 172.50 parts by weight of hydroxypropyl-v-cyclodextrin are introduced in step (i) of the method. 70. Pharmaceutical composition according to claim 68 or 69, which is characterized by the fact that 825 parts by volume of water are introduced into the stage (th) of the method. 70 71. Pharmaceutical composition according to any one of claims 59-70, which is characterized in that the stage of lyophilization (ii) includes: a) an initial stage of freezing, in which the complexation solution is brought to a temperature from about -40 °C to about -80 °C C for 2 to 4 hours; p) the primary stage of drying at a temperature of approximately -25 °C for approximately 80-90 hours; and c) a secondary drying stage at a temperature of about 30 °C for about 15-20 hours. 72. The pharmaceutical composition according to claim 71, which is characterized by the fact that stage (a) of lyophilization is carried out at a temperature of approximately -45 °C for approximately 3-4 hours. 73. Pharmaceutical composition according to claim 71 or 72, which differs in that stage (B) of lyophilization is carried out at a pressure of approximately 100 mTorr. 74. Pharmaceutical composition according to any one of claims 59-73, which is characterized by the fact that the stage of formulation (im) of the method includes mixing the complex with magnesium stearate and pressing into tablets. 75. Pharmaceutical composition according to claim 74, which is characterized by the fact that magnesium stearate is pre-mixed with sorbitol powder before mixing with the complex. c (8) « «c) сo «- d) - with . and? (ee) - (95) o 50 s» F) ime) 60 b5