CN1787810B

CN1787810B - Cladribine formulations for improved oral and transmucosal delivery

Info

Publication number: CN1787810B
Application number: CN2004800127149A
Authority: CN
Inventors: N·S·博多
Original assignee: Ares Trading SA
Current assignee: Ares Trading SA
Priority date: 2003-03-28
Filing date: 2004-03-26
Publication date: 2010-04-21
Anticipated expiration: 2024-03-26
Also published as: CN1787810A; UA81304C2; CN100408028C; CN1787809A; ZA200507939B; ES2349136T3; ZA200507935B

Abstract

Provided are compositions of cladribine and cyclodextrin which are especially suited for the oral administration of cladribine.

Description

The Cladribine formulations that per os that is used to improve and saturating mucosa are carried

Technical field

The present invention relates to make the compositions that comprises cladribine-cyclodextrin complexes of solid oral dosage form or saturating mucosa dosage form, also relate to the method for the oral and saturating mucosa bioavailability that strengthens cladribine.

Background technology

Cladribine is the unsettled medicine of a kind of acid, and it has chemical constitution as described below:

It is also referred to as 2-chloro-2 '-Deoxyadenosine or 2-CdA.

Cladribine is a kind of antimetabolite, can be used for the treatment of the lymphocytic hyperplasia disease.It has been used for the treatment of for example L1210 of experimental leukemia, is used for hairy cell leukemia and chronic lymphocytic leukemia and Walden Si Telunshi macroglobulinemia clinically.It is also as the medicine of immunosuppressant with treatment various autoimmune disease, described autoimmune disease (for example comprises rheumatoid arthritis, inflammatory bowel, Crohn disease, ulcerative colitis) and multiple sclerosis (for example see, J.Liliemark, Clin.Pharmacokinet, 32 (2): 120-131,1997).Also in experiment or studied it clinically, for example in lymphoma, Langerhans' cells histiocytosis, lupus erythematosus, chronic psoriasis en plaques, Sezary syndrome, Bing-Neel syndrome, recurrent glioma and solid tumor.

Owing to multiple reason, at first be patient's compliance, or because the consideration of cost or treatment aspect, the oral delivery medicine is through being often better than parenteral delivery.Peroral dosage form can reduce to medical personnel repeat go to a doctor or number of times is inculcated in injection discomfort or the prolongation relevant with some active medicine, strengthen patient's compliance thus.In the epoch that the health care cost increases gradually, become more important with relevant the reducing cost of using of mucosa per os or saturating with respect to the parenteral administration cost.The cost of parenteral administration is much higher, because need health care professional to use cladribine in health care person's place, this also comprises with this uses all relevant fringe costs.And in some cases, the consideration of treatment aspect for example needs through long time slow release cladribine, in fact can only be by the realization that transports of mucosa per os or saturating.

But up to now, per os and saturating mucosa carry cladribine to have following shortcoming: low bioavailability (is seen, for example, J.Liliemark etc., J.Clin.Oncol., 10 (10): 1514-1518,1992) and not good enough patient's differences (see, for example, J.Liliemark, Clin.Pharmacokinet, 32 (2): 120-131,1997).Also see the relatively poor absorption and the dependent unstability of pH (Arch.Immunol.et Therapie Exper., 42:13-15,1994) of report such as A.Tarasuik.

The ring-type oligosaccharide that the D-glucopyranose unit that cyclodextrin is connected by ring-type α-(1 → 4) is formed.Will have the unitary cyclodextrin of 6-8 respectively called after α-, β-and gamma-cyclodextrin.Unitary number has determined the size of conical cavity, and the latter can characterize cyclodextrin, and medicine can be wrapped into wherein, forms stabilized complex.The many α of cicada-, β-and gamma-cyclodextrin derivant, wherein one or more hydroxyls are replaced with ether or other group.Therefore, known these chemical compounds are complexing agents, have been used for pharmaceutical field in the past, to form inclusion complex with water-insoluble medicine, they are dissolved in the aqueous medium.

Recently, Schultz etc. have described the compound and solubilising cladribine with cyclodextrin at U.S. Patent number 6,194 among the 395B1.The inherent problem of described cladribine aqueous formulation before the patent of Schultz etc. has mainly solved is particularly for subcutaneous and intramuscular injection agent.Schultz etc. have been found that cladribine when preparing with cyclodextrin, not only can more be soluble in the aqueous medium significantly, and when combining with cyclodextrin, more stable to acid catalyzed hydrolysis.Back one is found for the preparation solid oral dosage form particularly useful, and wherein chemical compound can experience hydrolysis usually in the acid pH of gastric content.As if Schultz etc. do not describe any real work relevant with solid oral dosage form.In fact, they have only described a kind of method for preparing solid dosage forms, and this is a kind of melt extrusion method, wherein cladribine and cyclodextrin and other optional additive are mixed together, and heat up to fusion then.And 1mg to the 15mg cladribine of listing in this patent and the wide dosage range of 100mg to 500mg cyclodextrin show that with respect to the cladribine of specified rate in the solid oral dosage form, the concrete amount of the cyclodextrin that should exist is not vital.In fact, these dosage ranges have comprised many combinations, and it may be suitable as mixture, do not form but be suitable for complex.For example, the ratio of 1mg cladribine and 500mg cyclodextrin has comprised too many cyclodextrin, so medicine can not easily leave complex and realize its treatment function.On the other hand, the 15mg cladribine and only the 100mg cyclodextrin be not enough to the cladribine of compound this amount.

Though the patent of Schultz etc. has proposed by combining with cyclodextrin/the compound stability that improves the cladribine in the peroral dosage form, there is not to propose to improve the oral administration biaavailability of medicine by such mode; In fact, this patent does not have description or proposition enhancing or maximization method or the specially designed compositions that is used to realize this purpose from the bioavailability of the cladribine of the solid oral dosage form of cladribine and cyclodextrin.And, Schultz etc. do not have to propose to be used for the cladribine/cyclodextrin combination of mucosal administration, that is to say, when using, strengthen bioavailability of medicament to be intended to through nose, mouth, vagina or rectal cavity liner mucosa rather than through mouthful form of stomach administration, to say nothing of with this dosage form.

Many personnel after deliberation the dissolubility of certain drug in the water of the selected cyclodextrin that contains variable concentrations, can improve the dissolubility of this medicine with the concentration that confirm to improve cyclodextrin, be reported in the patent as for example Schultz etc. in selected temperature and pH level.Different personnel have also carried out phase solubility research, and to illustrate the person's character that complex forms, for example, whether cyclodextrin and medicine form 1: 1 complex or 1: 2 complex; See, for example, the U.S. Patent number 4 of Harada etc., 497,803, it relates to the inclusion complex of lankavacidin class antibiotic and cyclodextrin, U.S. Patent number 4 with Shinoda etc., 478,995, it relates to (2 '-benzyloxycarbonyl) phenyl trans-acid-addition salts of 4-guanidine radicals hexahydrotoluene formic acid esters and the complex of cyclodextrin.

At pharmaceutical field, in the solid dosage forms of using cyclodextrin raising medicine dissolution, use excessive cyclodextrin to become routine techniques, unless dissolubility can make excessive cyclodextrin cause too big dosage form.General knowledge shows; for solid oral dosage form, particularly sour unsettled medicine is cladribine for example, expects that excessive cyclodextrin can protect medicine in the sour environment of stomach; and carry it through intestinal wall/gastric mucosa ideally, still as being protected with the complex of cyclodextrin.In blood flow, away from the adverse effect of gastric acid, then expected drug can dissociate from complex, and realizes its treatment function.

Although instructions such as Schultz, cladribine-cyclodextrin complexes can improve the water solublity and the absolute acid stability of cladribine, the prior art is not pointed out, when in solid oral dosage form or saturating mucosa dosage form, using this complex, how with regard to bioavailability and patient's differences, to maximize or strengthen compound benefit.

Summary of the invention

Have been found that now, excessive cyclodextrin can suppress to comprise the absorption of the solid oral dosage form or the cladribine in the saturating mucosa dosage form of cladribine-cyclodextrin complexes, and the Peroral solid dosage form of saturated cladribine-cyclodextrin complexes or saturating mucosa dosage form can improve between the lower medicine patient of the bioavailability of oral and/or saturating mucosa and/or realization and/or the interior difference of patient.

The invention provides a kind of pharmaceutical composition, it comprises saturated cladribine-cyclodextrin complexes of making solid oral dosage form or saturating mucosa dosage form, is substantially devoid of to surpass the cyclodextrin that the amount that makes the cladribine in the complex maximizes required minimum.Of the present invention one concrete aspect, pharmaceutical composition comprises saturated cladribine-cyclodextrin complexes of making solid oral dosage form or saturating mucosa dosage form, is substantially devoid of to surpass the cyclodextrin that will all basically cladribines maintains minimum required in the complex.Said composition provides when the cladribine that has high heating power activated state when contacting with the mucosa (for the situation of saturating mucosa dosage form) of gastric mucosa (for the situation of peroral dosage form) or rectum, vagina, oral cavity or nose.

The present invention also provides the method that improves the oral or saturating mucosa bioavailability of cladribine, comprise object drug administration compositions to needs, wherein comprise saturated cladribine-cyclodextrin complexes of making solid oral dosage form or saturating mucosa dosage form, be substantially devoid of the cyclodextrin of the minimum required above the amount maximization that makes the cladribine in the complex.Of the present invention one concrete aspect, the compositions of using comprises saturated cladribine-cyclodextrin complexes of making solid oral dosage form or saturating mucosa dosage form, is substantially devoid of to surpass the cyclodextrin that will all basically cladribines maintains minimum required in the complex.

The present invention also provides and has improved from the cladribine of solid oral dosage form or saturating mucosa dosage form need be with the method for the bioavailability in the mammal of cladribine treatment, and this method comprises: (a) determine compound and described a selected amount of cladribine maintained the minimum of cyclodextrin required in the complex with a selected amount of cladribine; (b) cyclodextrin that surpasses described a selected amount of cladribine and described minimum is merged in aqueous medium; (c) from complex media, remove not compound cladribine; (d) from the solution that obtains, remove water, obtain exsiccant saturated cladribine-cyclodextrin complexes; (e) described exsiccant saturated cladribine-cyclodextrin complexes is mixed with solid oral dosage form or saturating mucosa dosage form, it is substantially devoid of and surpasses the cyclodextrin that the amount that makes the cladribine in the complex maximizes required minimum; (f) with this dosage form per os ground or saturating mucosa be administered to mammal.Of this method concrete aspect, step (e) comprises makes solid oral dosage form or saturating mucosa dosage form with described exsiccant saturated cladribine-cyclodextrin complexes, is substantially devoid of to surpass the cyclodextrin that will all basically cladribines maintains minimum required in the complex.

The present invention also provides and has treated mammiferous to using the method for the disease that cladribine responds by using compositions of the present invention.Also provide cladribine to be used for the application of administration with the pharmaceutical composition of the present invention of the bioavailability of the oral or saturating mucosa of reactive condition symptoms of treatment cladribine and enhancing cladribine in preparation.

In a specific embodiments, the invention provides a kind of new particularly advantageous cladribine: 1: 2 complex of gamma-cyclodextrin.In a relevant embodiment, provide 1: 1 cladribine: the cladribine of gamma-cyclodextrin complex and 1: 2: the mixture of gamma-cyclodextrin complex, wherein 1: 2 complex accounts for main.

Description of drawings

With reference to following detailed and accompanying drawing, can more easily understand more complete value of the present invention and its many attendant advantages, wherein:

Fig. 1 is the result's of phase solubility research a diagram, wherein use different cyclodextrin (CD) molar concentration to different cladribine molar concentration mappings, represent HP-with (◆), (■) representative has the HP-of the hydroxypropyl emthylcellulose of interpolation, and (▲) represents gamma-cyclodextrin.

Fig. 2 has shown the plasma profile of using the cladribine behind the 5mg single dose cladribine to Canis familiaris L., data show the standard deviation of mean concentration (pg/ml) ± 5-6 the animal/group of cladribine in blood plasma, to the time (hour) mapping, use following Cladribine formulations: (◇) intravenous (i.v.) injects; (*) the saturated cladribine gamma-cyclodextrin complex of sucking; The cladribine of sucking-HP-complex that (*) is saturated; Oral cladribine-gamma-cyclodextrin complex that (●) is saturated; (zero) oral capsule of the physical mixture of cladribine and 10 times of excessive gamma-cyclodextrins; () oral capsule of cladribine complex and 10 times of excessive gamma-cyclodextrins; Oral cladribine-HP-complex that (▲) is saturated; (△) oral capsule of the physical mixture of cladribine and 10 times of excessive HP-.

Fig. 3 has contrasted the plasma profile of using the cladribine behind the cladribine of 5mg single dose to Canis familiaris L., data show the standard deviation of mean concentration (pg/ml) ± 5-6 animal/group, to the time (hour) mapping, use following Cladribine formulations: (◆) intravenous (i.v.) injects, oral cladribine-HP-complex that oral cladribine-gamma-cyclodextrin complex that (●) is saturated and (▲) are saturated.

Fig. 4 has contrasted the plasma profile to the cladribine behind the cladribine of Canis familiaris L. oral administration 5mg single dose, data show the standard deviation of mean concentration (pg/ml) ± 5-6 animal/group, to the time (hour) mapping, use following Cladribine formulations: oral cladribine-gamma-cyclodextrin complex that (●) is saturated; (zero) oral capsule of the physical mixture of cladribine and 10 times of excessive gamma-cyclodextrins; () oral capsule of cladribine complex and 10 times of excessive gamma-cyclodextrins.

Fig. 5 has contrasted the plasma profile to the cladribine behind the cladribine of Canis familiaris L. oral administration 5mg single dose, data show the standard deviation of mean concentration (pg/ml) ± 5-6 animal/group, to the time (hour) mapping, use following Cladribine formulations: oral cladribine-HP-complex that (▲) is saturated; (△) oral capsule of the physical mixture of cladribine and 10 times of excessive HP-.

Fig. 6 illustrated and used about Fig. 2 behind the described various preparations, cladribine in the group of 5-6 Canis familiaris L. to the time (hour) average accumulated area under curve (AUC) (pgx h/mL) that mapping obtains, wherein each sign is as shown in Figure 2.

Detailed Description Of The Invention

In the present specification and claims, adopt following definition and general description.

The patent of quoting in this article, disclosed application and scientific and technical literature have been set up those skilled in the art's knowledge, and they are all whole incorporated by reference here, and its degree is with indivedual incorporated by reference identical especially. If there be any conflict in any document of quoting here with the concrete instruction of this specification, should be as the criterion with the latter. Similarly, if there be any conflict in the definition to word or phrase that this area is understood with the word of the concrete instruction of this specification or the definition of phrase, should be as the criterion with the latter.

As used herein term " compound " refers to inclusion complex, and wherein the hydrophobic part of Cladribine molecule (nitrogenous ring system) inserts in the hydrophobic cavity of cyclodextrin molecular.

As used herein, no matter be in the transition phrase or in the claim main body, term " comprises " and " comprising " all should be understood to have open implication. That is to say that this term should be understood to " have at least " or " comprising at least " synonym with phrase. When using in the context in method, term " comprises " and refers to that the method comprises described step at least, but can also comprise other step. When using in the context at composition, term " comprises " and refers to that said composition comprises described feature or component at least, but also can comprise further feature or component.

Term " basically by ... form that " have semi-enclosed implication, that is to say, they do not allow to comprise step or feature or the component of the essential characteristic that can change in fact method or composition; For example, can significantly disturb step or feature or the component of the destination properties of composition as herein described, namely the method or composition are limited to the step of appointment or material and can be from not affecting in essence those of basic new feature of the present invention. Basic new feature here is, saturated Cladribine-cyclodextrin complexes in solid oral dosage form or saturating mucous membrane formulation is provided, it is substantially devoid of the cyclodextrin that surpasses the required minimum of the amount maximization make the Cladribine in the compound, thereby difference between the bioavilability of improvement and/or lower patient can be provided after using. In a specific embodiments of the present invention, basic new feature here is, saturated Cladribine-cyclodextrin complexes in solid oral dosage form or saturating mucous membrane formulation is provided, it is substantially devoid of and surpasses the cyclodextrin that will be basically all Cladribines maintain minimum required in the compound, can provide between the bioavilability of special enhancing and/or low patient after using and/or otherness in the low patient.

Term " by ... form " and " compositions " be the term that seals, only allow to comprise described step or feature or component.

As used herein, singulative " ", " a kind of " and " being somebody's turn to do " also comprise the plural form of the term of their indications particularly, unless context has clearly explanation in addition.

Term " pact " be used in reference to about, neighbouring, rough in this article or about." pact " and numerical range are united when using when term, it by with border extension to more than the described numerical value and with this scope of modification of getting off.Usually, term " pact " or " approximately " be used in this article modifying more than the described value numerical value and following 20% deviation.

When with cyclodextrin in the complex of cladribine unite when using, term " saturated " be meant that this complex is that cladribine is saturated, that is to say that this complex contains can be under the compound condition of using and the compound maximum cladribine of the cyclodextrin of specified rate.Phase solubility research can be used to provide this information, as (following also described in more detail compound condition) hereinafter in greater detail.Perhaps, can obtain saturated complex, by in the aqueous solution of selected cyclodextrin, adding cladribine simply, up to forming (not compound cladribine) precipitation empirically; Finally, remove precipitation, lyophilizing solution obtains exsiccant saturated complex.

Statement " basically ", as in " being substantially devoid of " or " all basically ", be meant the accurate Calculation amount 20% in.In the situation that statement " is substantially devoid of and surpasses the cyclodextrin that will all basically cladribines maintains minimum required in the complex ", from as below the phase solubility research explained in more detail, can obtain cladribine is maintained the minimum of cyclodextrin required in the complex.The actual amount of cyclodextrin should be in this minima ± 20%, preferably in this minima ± 10%, even more preferably in this minima ± 5%, and should with at least 90% or more, preferably at least 95% or more medicine maintain in the complex.On the other hand, when using statement " to be substantially devoid of and to surpass the cyclodextrin that the amount that makes the cladribine in the complex maximizes required minimum ", can use the cyclodextrin that is lower than aforementioned quantities, may there be the cladribine of more substantial not complex form in the result in dosage form.Carry out compound reaction and/or compound by the cyclodextrin solution that uses lower concentration, this phenomenon can take place by carrying out in the upper limit of following temperature range.But, thereby think and use enough cyclodextrin all basically cladribines to be maintained the amount of compound cladribine is minimized is useful especially.

Term " patient's differences " is the difference of pointing between the patient of its drug administration.Term " diversity in the patient " is meant the difference that same patient is experienced when at different time administrations.

As used herein, the numerical range of described variable is intended to show, uses the variable that equates with the interior any value of this scope, can realize the present invention.Thereby for discrete in essence variable, this variable can equal any integer value in this scope, comprises the end value of this scope.Similarly, for successive variable in essence, this variable can equal any real number value in this numerical range, comprises the end value of this scope.As an example, be described as variable with the value between the 0-2, for discrete in essence variable, can be 0,1 or 2, for successive variable in essence, then can be 0.0,0.1,0.01,0.001 or any other real number value.

In description and claim, singulative comprises plural form, unless context has clearly explanation in addition.As used herein, unless otherwise specified, " comprising " implication of the word of use " or " have " and/or ", rather than " eliminating " implication of " not being/be exactly ".

Technology used herein and scientific and technical terminology have the implication of those skilled in the art in the invention's common sense, unless otherwise defined.Here learn and material with reference to the whole bag of tricks known to those skilled in the art.The normative document of describing pharmacological ultimate principle comprises the The Pharmacological Basis of Therapeutics of Goodman and Gilman, the 10th edition, and McGraw Hill Companies Inc., New York (2001).

Hereinafter in more detail with reference to specific embodiments of the present invention.Although will describe the present invention, should be understood that unintentionally the present invention to be limited to these specific embodiments in conjunction with these specific embodiments.On the contrary, be intended to cover and be included in substituting in the defined the spirit and scope of the present invention of appended claims, improvement and equivalent.In the following description, described numerous details, understood the present invention with thorough.Do not have in these details partly or entirely, also can realize the present invention.In other situation,, do not describe well-known technological operation in detail in order not make unnecessary obscure of the present invention.

The invention provides the method for the pharmaceutical composition of the pharmacokinetic property that compositions and preparation and use be used to realize ideal.Such source is from following discovery: when being presented to will be through (stomach, nose, rectum, the oral cavity, the Sublingual or vagina) mucosa of its absorption the time, wherein cladribine is in the cyclodextrin of its hyperpyrexia mechanical state and the solution of cladribine absorbs with the cladribine that improves, and is reflected as higher bioavailability and/or lower patient's differences.

Do not wish so to limit the present invention, (for example suppose by dissolving, by with fluid for example body fluid contact), the dry compositions that does not contain the saturated cladribine-cyclodextrin complexes of excessive cyclodextrin can form local saturated cladribine solution, wherein cladribine is in high heating power activity (HTA) state, thereby helps absorbing.Cladribine has quite low (although not being insignificant) intrinsic water solublity.The free cladribine that forms from complex dissociation in saturated aqueous solution shows more stable activity level, if there is excessive cyclodextrin, cladribine can be by seeking bigger stability with cyclodextrin is compound again.By the amount of control loop dextrin, make this dosage form be substantially devoid of to surpass the cyclodextrin that cladribine is maintained amount required in the complex, the cladribine in the local saturated solution can not easily combine with cyclodextrin again.Therefore, this cladribine can be sought lower thermodynamic activity/bigger stability status by the absorption through gastric mucosa (for the situation of solid oral dosage form) or mucosa per nasal, the oral cavity, vagina or rectum (for the situation of saturating mucosa dosage form).Except that other guide, confirmed especially that hereinafter this scheme can improve bioavailability, may be by avoiding or minimizing because the inhibition that exists excessive cyclodextrin to cause to the cladribine absorption.Having in the presence of a large amount of excessive cyclodextrin, the cladribine in the expection solution can combine again with cyclodextrin.This can not realize best bioavailability, because if cladribine will be realized its treatment function, this medicine must move out from the complex of sealing it.

In view of foregoing, obviously to produce the optimal drug compositions of Peroral solid dosage form or saturating mucosa dosage form, these dosage forms should make when solid dosage forms with can discharge local saturated cladribine solution after the body fluid at mucosa place contacts, wherein cladribine is in its HTA state.In order so local saturated solution to be provided in vivo, importantly at first to identify the cladribine that will be used for solid dosage forms and the optimal proportion of cyclodextrin, this ratio is called the HTA ratio in this article.In sucking the situation of dosage form, by saturated complex is dissolved in the water of minimum, made highly spissated solution, place the oral cavity can realize identical effect this solution.

Determine the HTA ratio empirically, and it be accredited as the ratio of cladribine and specific cyclodextrin, its correspondence can with the cladribine of the compound maximum of cyclodextrin of specified rate.Use experience method (for example phase solubility research) detect can with the saturated concentration of the dissolved cladribine of cyclodextrin solution of variable concentrations, can determine the HTA ratio.Thus, this method can identify the concentration when forming saturated cladribine-cyclodextrin complexes.The mol ratio that should be noted that the some representative on the phase solubility figure has shown that the cyclodextrin of how many moles is under specified criteria this medicine to be maintained minimum required in the complex; Can convert it into weight ratio then.For example, if phase solubility figure shows, basically all cladribines are maintained needs 9 moles of given cyclodextrin in the saturated complex, by with its molecular weight of molal quantity X of cladribine with its molecular weight of molal quantity X of cyclodextrin, can obtain ratio so as the product of suitable optimized weight ratio.Phase solubility research also provides the information about the character of the cladribine-cyclodextrin complexes that forms, for example, this complex is 1: 1 complex (1 molecular medicine and 1 molecule cyclodextrin are compound) or 1: 2 complex (1 molecular medicine and 2 molecule cyclodextrin are compound).

According to the present invention, can use cyclodextrin or cladribine to begin as fixed variable, to wherein adding excessive another kind, differentiating different HTA data point (indicating saturated cladribine-cyclodextrin complexes), and draw the HTA line that obtains.Typically, find to promote in the aqueous solution of the cyclodextrin that contains concentration known, to add cladribine under the condition of complex formation by rule of thumb.Concentrated solution, for example, the solution of the solution of about 27% gamma-cyclodextrin and about 40% HP-is useful especially in one embodiment.Usually, (about at the most 50 ℃ or even up to 60 ℃) carries out compound in room temperature or under mild heat.Remove the excessive cladribine that may exist then, detect the cladribine concentration in the complex subsequently.The concentration that records is being represented the cladribine saturated concentration for given cyclodextrin concentration.For the cyclodextrin of different concentration known, repeat this method, up to obtaining several data points.Each data point is being represented the saturated concentration of the cladribine in the cyclodextrin that is dissolved in concentration known.Draw with data point then, demonstrate saturated concentration at the cladribine of the different cyclodextrin concentration that uses.This figure is phase solubility figure, and it can be used to determine under given compound condition, for any specifically saturation capacity of the cladribine of concentration of the cyclodextrin that is used to form saturated cladribine-cyclodextrin complexes.

Those skilled in the art can understand, by multiple alternative method, can identify the concentration (and HTA ratio) that forms saturated cladribine-cyclodextrin complexes.Therefore, known in the art be applicable to differentiate these concentration any method all within the scope of the invention.

Have been found that ideal pharmacological property (between the bioavailability of improvement and/or lower patient and/or patient in difference) is relevant with inclusion complex of the present invention.

Cyclodextrin in the scope of the invention comprise natural cyclodextrin α-, β-and gamma-cyclodextrin and derivant thereof, more specifically, in derivant, one or more hydroxyls are substituted, and are for example replaced by alkyl, hydroxyalkyl, carboxyalkyl, alkyl-carbonyl, carboxyl alkoxyalkyl, alkyl-carbonyl oxyalkyl, alkoxy carbonyl alkyl or hydroxyl-(single or many alkoxyls) alkyl; And wherein each alkyl or alkylidene preferably contain maximum 6 carbon.Substituted cyclodextrin generally can obtain with different substitution values, for example, and from 1 to 14, preferably from 4 to 7; Substitution value is the number substituent on a rough average on the cyclodextrin molecular, for example, under the situation of HP-molecule, is the roughly number of hydroxypropyl, and all such variations all within the scope of the invention.Operable in the present invention substituted cyclodextrin comprises polyethers, and for example, as U.S. Patent number 3,459,731 is described.Other example of substituted cyclodextrin comprises ether, and the hydrogen of wherein one or more cyclodextrin hydroxyls is replaced with C _1-6Alkyl, hydroxyl-C _1-6Alkyl, carboxyl-C _1-6Alkyl or C _1-6Alkoxy carbonyl-C _1-6Alkyl, or its compound ether.More specifically, so substituted cyclodextrin is an ether, and the hydrogen of wherein one or more cyclodextrin hydroxyls is replaced with C _1-3Alkyl, hydroxyl-C _2-4Alkyl or carboxyl-C _1-2Alkyl, or more specifically substituted by methyl, ethyl, ethoxy, hydroxypropyl, hydroxyl butyl, carboxymethyl or carboxyethyl.Term " C _1-6Alkyl " is intended to comprise saturated alkyl straight chain with 1-6 carbon atom and that side chain is arranged, for example methyl, ethyl, 1-Methylethyl, 1,1-dimethyl ethyl, propyl group, 2-methyl-propyl, butyl, amyl group, hexyl etc.Other cyclodextrin that is used for this paper purposes comprises glucose group-beta-cyclodextrin and malt sugar group-beta-cyclodextrin.Useful especially in the present invention is beta-schardinger dextrin-ether, and for example M.Nogradi (1984) is at Cyclodextrins of the Future, Vol.9, No.8, p.577-578 the DM-described in, methylated at random beta-schardinger dextrin-, and polyethers, HP-for example, hydroxyethyl-, hydroxypropyl-gamma-cyclodextrin, and ethoxy-gamma-cyclodextrin, and sulfur butyl ether, particularly beta-schardinger dextrin-sulfur butyl ether.Except simple cyclodextrin, can also use the cyclodextrin and the cyclodextrin of side chain.Other cyclodextrin is documented in, for example, and Chemical and PharmaceuticalBulletin 28:1552-1558 (1980); Yakugyo Jiho No.6452 (28 March1983); Angew.Chem.Int.Ed.Engl.19:344-362 (1980); U.S. Patent number 3,459,731 and 4,535,152; European patent number EP 0 149 197A and EP 0197 571A; Pct international patent publication number WO90/12035; With the open GB2 of British patent, 189,245.Described and be used for according to the cyclodextrin of compositions of the present invention and provide other document of preparation, purification and the analysis guide of cyclodextrin to comprise following: Jozsef Szejtli, the Cyclodextrin Technology of Kluwer Academic Publishers (1988) in Cyclodextrin inPharmaceuticals one chapter; M.L.Bender etc., Springer-Verlag, the Cyclodextrin Chemistry of Berlin (1978); Advances in Carbohydrate Chemistry, the 12nd volume, M.L.Wolfrom compiles, Academic Press, New York, " The SchardingerDextrins " chapter of Dexter French, 189-260 page or leaf; J.Szejtli, Akademiai Kiado, Budapest, the Cyclodextrin and their Inclusioncomplexes of Hungary (1982); I.Tabushi, Acc.Chem.Research, 1982,15,66-72 page or leaf; W.Sanger, Angewandte Chemie, 92, the 343-361 pages or leaves (1981); A.P.Croft etc., Tetrahedron, 39, the 1417-1474 pages or leaves (1983); Irie etc., Pharmaceutical Research, 5, the 713-716 pages or leaves (1988); Pitha etc., Iht.J.Pharm.29,73 (1986); U.S. Patent number 4,659,696 and 4,383,992; German patent DE 3,118, and 218 and DE-3,317,064; With european patent number EP 0 094157A.Describe β-and the patent of the hydroxyalkylation derivant of gamma-cyclodextrin comprise the U.S. Patent number 4,596,795 and 4,727,064 of Pitha, the U.S. Patent number 6,407,079 of the U.S. Patent number 4,764,604,4,870,060 of M ü ller and M ü ller etc.

Making us interested being used for especially comprises with the compound cyclodextrin of cladribine: gamma-cyclodextrin β-and hydroxyalkyl (for example ethoxy or the hydroxypropyl) derivant of gamma-cyclodextrin; β-or carboxyalkyl (for example carboxymethyl or the carboxyethyl) derivant of gamma-cyclodextrin; Beta-schardinger dextrin-sulfur butyl ether; DM-; Methylated at random beta-schardinger dextrin-.2-HP-(HP β CD), 2-hydroxypropyl-gamma-cyclodextrin (HP γ CD), methylated at random beta-schardinger dextrin-, DM-, beta-schardinger dextrin-sulfur butyl ether, carboxymethyl-beta-cyclodextrin (CM β CD), carboxymethyl-gamma-cyclodextrin (CM γ CD) and gamma-cyclodextrin (γ CD) they itself are particularly advantageous, particularly gamma-cyclodextrin and HP-, the most particularly gamma-cyclodextrin.

Explain and example as this paper, can help under the condition that complex forms, in liquid environment, prepare the compositions that is used for saturated cladribine-cyclodextrin complexes of the present invention.The flowing product that obtains can be changed into subsequently and be applicable to the dried forms of using as Peroral solid dosage form or saturating mucosa dosage form.

The technical staff can understand that this area can obtain multiple preparation method for compositions as herein described.A kind of available method of this paper institute example comprises the steps: cladribine is mixed in the cyclodextrin aqueous solution, complex media is maintained room temperature, stir about 6 was to about 24 hours, promptly be enough to realize balance time, the not compound cladribine that separation may exist (for example, by filtration or centrifugal), lyophilizing or lyophilization saturated solution form the saturated cladribine-cyclodextrin complexes mixture of solid.

Lyophilization is also referred to as lyophilizing, is made up of 3 foundation phases: at first being the freezing stage, is drying stage first then, is the redrying stage at last.The following examples 2 provide the described batch of freeze dried details of formula.By following whole incorporated by reference and Xiaolin (Charlie) Tang and the Michael J.Pikal in PharmaceuticalResearch that rely at this paper, the 21st volume, the 2nd phase, in February, 2004, the described principle of 191-200 can further be optimized this method.

Can randomly comprise one or more excipient or other pharmaceutically inert component according to pharmaceutical composition of the present invention.But one of advantage of the present invention is, uses to form and the excipient of the necessary minimum of production particular form (for example tablet or patch), just can prepare cladribine medicament forms as described herein.Can never can influence in those excipient that cladribine, cyclodextrin or complex form and select.

Randomly prepare dosage form together and (see Handbook of PharmaceuticalExcipients at the well-known arbitrarily pharmaceutically acceptable carrier of pharmaceutically acceptable medium neutralization, diluent, binding agent, lubricant, disintegrating agent, cleanser, correctives, coloring agent and excipient, Marcel Dekker Inc., New York and Basel (1998); Volumes such as Lachman, The Theory and Practice of Industrial Pharmacy, the 3rd edition, (1986); Lieberman etc., Eds.Pharmaceutical Dosage Forms, Marcel Dekker Inc., New York and Basel (1989); With The Handbookof Pharmaceutical Excipients, the 3rd edition, American PharmaceuticalAssociation and Pharmaceutical Press, 2000); Also see Rermington ' sPharmaceutical Sciences, the 18th edition, Gennaro, Mack Publishing Co., Easton, PA (1990) and Remington:The Science and Practice ofPharmacy, Lippincott, Williams ﹠amp; Wilkins, (1995)).Simple Peroral solid dosage form or saturating mucosa dosage form are by forming with the saturated cladribine-cyclodextrin complexes of (for example about 1% weight) suitable bonding or lubricant (for example magnesium stearate) compacting in a small amount.

In specific embodiment, saturated cladribine-cyclodextrin complexes is used cladribine with being used for mucosa ground or per os.

As used herein, " mucosa " refers to the epithelial membrane of nose, oral cavity, vagina and rectal cavity liner, and the epithelial membrane (gastric mucosa) of gastric lining.As used herein, the use interchangeably of mucosa mucosa and saturating.The carrying method and the form of saturating mucosa are well-known in the art.They comprise that suck and the tablet Sublingual, lozenge, adhesive patch, gel, solution or spray (powder, liquid or aerosol) and suppository or foam (be used for rectum or the using of vagina).The carrying method of saturating mucosa and form do not comprise method and the form that is used for the oral administration use, and the latter is intended to be swallowed, and is called peroral dosage form in this article simply, although they finally can carry medicine by gastric mucosa.When the form of saturating mucosa is liquid, can be by obtaining in the water that saturated complex is dissolved in minimum, for example the saturated complex with 500mg and HP β CD is dissolved in (50% w/w solution) in the 0.5ml water, or the γ CD complex that 500mg is saturated is dissolved in the 1.0ml water.Can be with in several such drips of solution entrance cavities, and kept there about 2 minutes, to carry out the absorption of oral transmucosal.Yet the saturating mucosa dosage form of solid generally is better than liquid form.

In some cases, improve dissolubility or strengthen infiltration by adding different excipient and additive, for example by modifying microenvironment, or improve contact between induction system and the mucosal tissue by the excipient that add to adhere to mucosa, can further promote absorption oral administration or mucosa.

By sucking between the lower gum and the oral mucous membrane relative that dosage unit places the individuality of accepting Drug therapy, can realize the drug conveying in oral cavity with it.Can use and be applicable to excipient or the medium of sucking drug administration, comprise so arbitrarily material known in the art, for example, nontoxic and harmful interactional liquid arbitrarily, gel, solvent, liquid diluent, solubilizing agent etc. do not take place with other component of compositions.Solid dosage unit is made and can be dissolved gradually through the preset time stage,, the cladribine through mucous membrane is absorbed, wherein should provide drug conveying in the time period whole basically in saliva of buccal cavity, to generate saturated basically drug solution.Suck dosage unit and can also comprise the lubricant that can promote production, for example, magnesium stearate etc.Other component of sucking in the dosage unit be can be included in and correctives, penetration enhancers, diluent, binding agent etc. included but not limited to.The remainder of sucking dosage unit can comprise bioerodible polymeric carrier and any excipient that may need, for example binding agent, disintegrating agent, lubricant, diluent, correctives, coloring agent etc., and/or other active medicine.

Suck carrier and can comprise having viscous enough polymer,, promptly during this period cladribine is transported to the time period of oral mucosa to guarantee that this dosage unit can adhere to oral mucosa and continue the essential time period.In addition, polymeric carrier is " biological erosion is separated " gradually, i.e. this polymer speed hydrolysis to be scheduled to behind contact wetting.Can use pharmaceutically acceptable, that suitable adhesiveness and ideal drug release characteristics can be provided any polymeric carrier, and with the cladribine that will use with to suck any other composition that may exist in the dosage unit compatible.Usually, polymeric carrier comprises wet lip-deep hydrophilic (the water miscible and water-swellable) polymer that can adhere to oral mucosa.The example of polymeric carrier used herein comprises acrylate copolymer and copolymer, for example, is called those of " carbomer ", for example,

Other suitable polymers includes but not limited to polyvinyl alcohol, polyethylene glycol oxide (for example, the Sentry of hydrolysis

Polyacrylate (for example,

Polyvinyl and copolymer, polyvinylpyrrolidone, glucosan, guar gum, pectin, starch and cellulosic polymer, for example hydroxypropyl emthylcellulose (for example, Hydroxypropyl cellulose (for example, Hydroxypropylcelluloether ether, hydroxyethyl-cellulose, sodium carboxymethyl cellulose, methylcellulose, ethyl cellulose, acetic acid-O-phthalic acid cellulose, cellulose acetate butyrate (CAB) etc.Dosage unit only needs to contain saturated cladribine-cyclodextrin complexes.But in some cases, it can comprise one or more aforesaid carriers and/or one or more other components ideally.For example, can comprise the technical process that lubricant is conveniently produced this dosage unit; Lubricant can also be optimized erosion and separate speed and medicine outflow.If there is lubricant, its magnitude is that 0.01 weight % of dosage unit is to about 2 weight %, preferred about 0.01 weight % to 1.0 weight %.Examples of suitable lubricants includes but not limited to magnesium stearate, calcium stearate, stearic acid, sodium stearyl fumarate, Talcum, hydrogenated vegetable oil and Polyethylene Glycol.

According to the present invention, can also use saturated cladribine-cyclodextrin complexes with the suppository or the form of foam that are used for vagina or rectal administration.By well-known method, can prepare these compositionss, for example, in the situation of suppository, by saturated complex is mixed with suitable non-irritating excipient or binding agent, the latter is solid at normal temperatures, but is liquid under vagina or rectal temperature, therefore can in vagina or rectum, fuse, discharge medicine.Such material comprises cocoa butter and Polyethylene Glycol.Traditional binding agent and carrier comprise, for example, and poly alkylene glycol or triglyceride [for example PEG 1000 (96%) and PEG 4000 (4%)].From containing the mixture of 0.5 w/w % of having an appointment, can form such suppository to about 10 w/w %, preferred about 1 w/w % to the active component of about 2 w/w % scopes.

Use for intranasal, can use the powder spray agent or the ointment of saturated complex, preferred powder type.

And, for human, suck dosage form, particularly buccal tablet or wafer or dish advantageously have about 15-30 minute disintegration time, or buccal bioadhesive tablet (its Chinese medicine only discharges from that side that adheres on the oral mucosa, and opposite side is an impermeable), be interesting.The using of oral cavity can be utilized the invention of the Nagai of record in this paper whole incorporated by reference and the U.S. Patent number 4,226,848 and 4,250,163 that relies on etc.Thereby, purposes for this paper, can prepare the tablet that can adhere to oral mucosa, it comprises: (a) water-swellable with the polymeric matrices that can adhere to mucosa, it comprises about 50% cellulose ether and about 50% acrylate homopolymer or copolymer or its pharmaceutically acceptable salt to about 95% weight to about 95% weight, (b) be dispersed in the cladribine of Sq wherein, as with the saturated complex of 2-HP-or gamma-cyclodextrin.It is desirable to, for storage stability, this tablet is anhydrous.

Method as herein described and pharmaceutical composition can be for providing new form of therapy with the patient's of cladribine treatment treatment.As indicated in this paper, the invention solves the relatively poor bioavailability problem that is accompanied by oral cladribine traditionally.Perhaps, by using the transporting pattern of mucosa, can avoid a mouthful stomach approach fully.

Compositions of the present invention is specially adapted to the therapy as any cladribine reactive disorder of treatment.Several morbid states (seeing below) that cladribine is responded have fully been put down in writing in the document.For target disease state arbitrarily, use the cladribine-cyclodextrin complexes (for example, effectively measuring) of the optimization of effective dose for treatment multiple sclerosis, rheumatoid arthritis or leukemia.

Term " treatment effective dose " or " effective dose " are used to be illustrated in the treatment of the dosage that can realize the therapeutic outcome pursued effectively.Effective dosage comprises following treatment of diseases effective dose in the described in the literature treatment: hairy cell leukemia (0.09mg/kg/ days, carried out 7 days), multiple sclerosis (about 0.04 to about 1.0mg/kg/ day (seeing U.S. Patent number 5,506,214)); Other disease is also seen U.S. Patent number 5,106,837 (oneself soup-dissolving courage and uprightness anemias); 5,310,732 (inflammatory bowel); 5,401,724 (rheumatoid arthritiss); 5,424,296 (pernicious astrocytomas); 5,510,336 (histiocytosiss); 5,401,724 (chronic myelogenous leukemias); With 6,239,118 (atherosclerosiss).

In addition, different dosage and dosage regimen have been put down in writing in the literature, are used for the treatment of multiple sclerosis; See, for example: Romine etc., Proceedings of the Association ofAmerican Physicians, the 111st volume, the 1st phase, 35-44 (1999); Selby etc., The Canadian Journal of Neurological Sciences, 25,295-299 (1998); Tortorella etc., Current Opinion in InvestigationalDrugs, 2 (12), 1751-1756 (2001); Rice etc., Neurology, 54,1145-1155 (2000); With Karlsson etc., British Journal ofHaematology, 116,538-548 (2002); Their all whole here incorporated by reference and dependences.

And, should consider to instruct in the document route of administration for the treatment of effective dose.Although compositions of the present invention can be optimized the bioavailability of cladribine behind per os or saturating mucosal administration, be understood that, even best bioavailability from oral or saturating mucosa dosage form, can not the expectation meeting near use the bioavailability that the back obtains at intravenous, particularly at early time point; See, for example, Fig. 3 hereinafter.Thereby, often need increase to the dosage that intravenous is used recommendation, to reach the suitable dose that is integrated into solid oral dosage form or saturating mucosa dosage form.Think at present, for the treatment of multiple sclerosis, use the 10mg cladribine of the saturated cladribine-cyclodextrin complexes of the once conduct in this solid dosage forms every day, continue 5-7 days in first month, repeated other 5-7 days at the second month, do not treat in 10 months then.Perhaps, use 10mg dosage every day one time for the patient, every month 5-7 days, continue 6 months, do not treat in 18 months then.For other drug administration information, also see U.S. Provisional Patent Application number [IVAX0021-P-USA/ attorney docket 033935-011], with U.S. Provisional Patent Application number [IVAX0022-P-USA/ attorney docket 033935-012], the title of the two all is " Cladribine Regimen for Treating Multiple Sclerosis ", submit on March 25th, 2004, they are whole here incorporated by reference.

And the technical staff can understand, by fine setting and/or by using according to cladribine of the present invention with other active component, can reduce or improve the treatment effective dose of the cladribine of using in this article.Therefore, the invention provides administration and the method that adapts at given mammiferous concrete condition of making.Can easily determine the treatment effective dose, for example,, and progressively increase, estimate beneficial effect simultaneously empirically from relative a small amount of.

As in the previous paragraph, use, can be accompanied by and use other active component that one or more are used for the treatment of the reactive disease of cladribine according to cladribine of the present invention.With with every kind of other active component and the disease of treatment route of administration, dosage and frequency of adapting, use other active component.For example, in the treatment of multiple sclerosis, other useful medicine comprises interferon beta It is identical with the naturally occurring albumen of finding in human body; The glatiramer acetate Be the aminoacid GLAT with chain (polymer); Natalizumab

It is a kind of monoclonal antibody; Alemtuzumab Humanized anti-CD 52 monoclonal antibody; 4-aminopyridine (being also referred to as 4-AP and Fampridine), promptly a kind of can block nerves the medicine of potassium channel in the unit; And amantadine, promptly a kind ofly can improve muscle control and reduce muscle rigidity and be used for alleviating the antiviral drugs of the fatigue symptom of multiple sclerosis, for this purpose, can also use pemoline With L-carnitine (a kind of medical herbs goods).In the treatment of hairy cell leukemia, other active component can comprise interferon-ALPHA, pentostatin, fludarabine, Mabthera (a kind of resisting-CD 20 monoclonal antibodies) and anti--CD22 recombinant immunotoxin BL 22; Other additional active component may be suitable in the leukemia of other type.In the treatment of rheumatoid arthritis, can select many other active component.They comprise NSAIDS (NSAID (non-steroidal anti-inflammatory drug)), and it has 3 types: Salicylate is aspirin for example, and traditional NSAIDS is ibuprofen and indomethacin and cox 2 inhibitor celecoxib for example for example Rofecoxib Meloxicam

Valdecoxib

Lumiracoxib

And etoricoxib

Can comprise DMARDS, glucocorticoid, the analgesic of biological response modifier and non--NSAID with the other medicines that are used for the treatment of rheumatoid arthritis that the present invention unites use.DMARDS is a disease modulability antirheumatic, and it comprises methotrexate, chloroquine, leflunomide Sulfasalazine, gold, penicillamine, ciclosporin, methyl cyclophosphamide and azathioprine.Glucocorticoid comprises dexamethasone, prednisolone, triamcinolone and many other medicines.Biological response modifier (it can recover the immune ability that fights back the disease) comprises Embrel A kind of tumor necrosis factor inhibitors, infliximab It also is a kind of anti-TNF medicine, Antril (Synergen)

Be a kind of optionally IL-1 blocker and

Be a kind of human monoclonal antibodies, it is another kind of anti-TNF medicine.The analgesic of non--NSAID comprises acetaminophen and narcotic analgesic for example hydrocodone, oxycodone and dextropropoxyphene.Generally speaking, those medicines that its mechanism of action is different with cladribine are useful especially for treating together with cladribine compositions as herein described.Can with the per os of single dosage form or saturating mucosal administration approach effectively and those medicines compatible with cladribine complex of the present invention be integrated in this dosage form; In addition, they are certainly used dividually with the amount, frequency and the route of administration that are fit to them.

As used herein, " treatment " refer to, with not according to the present invention the symptom of the individuality of treatment compare, alleviate, prevent, stop the development of having used the symptom in the individuality of The compounds of this invention to it, control, alleviate and/or reverse its symptom.The practitioner can understand, determines follow-up treatment according to skilled operator's (doctor or veterinary) continuous clinical evaluation, uses complex as herein described, compositions, dosage form and method.Such evaluation can assist and inform whether checking increases, reduces or continue specific therapeutic dose and/or change administering mode.

Method of the present invention is intended to be used for can be from any object/patient of method benefit of the present invention.Thereby according to the present invention, term " object " and " patient " comprise people and inhuman object, particularly domestic animal.

Any suitable material known to the skilled and/or method may be used to implement the present invention.But, preferable material and method have been described.Except as otherwise noted, material of mentioning among explanation below and the embodiment and reagent etc. can obtain from commercial source.

The following examples are intended to further specify some preferred embodiment of the present invention, rather than restrictive.Only use normal experiment, those skilled in the art will appreciate that the many equivalents that maybe can determine concrete material as herein described and method.

Embodiment

Embodiment 1 phase solubility research

Followingly carry out phase solubility research.Excessive cladribine is added in the cyclodextrin solution of the gamma-cyclodextrin (γ CD) of variable concentrations or HP-(HP β CD), compound as carrying out as described in the following examples 2.In addition, in one group of experiment, studied hydroxypropyl emthylcellulose (HPMC) to compound influence.Remove unnecessary undissolved cladribine by filtering.Detect the amount of the cladribine in the composite solution, obtain data point.Use the cyclodextrin of different concentration known to repeat this method, up to obtaining several data points.Draw with these data points then, each data point representative can with the maximum of the compound cladribine of cyclodextrin of specific concentrations, i.e. a kind of saturated cladribine-cyclodextrin complexes of each some representative.Represent the HTA ratio by the point on the line of data point generation.A kind of specific, unique saturated cladribine-cyclodextrin complexes is represented in arbitrfary point on this line.Those skilled in the art will appreciate that,, can obtain identical result if excessive cyclodextrin is joined in the cladribine solution of concentration known.

As an example, prepared the cyclodextrin solution of variable concentrations, and by excessive cladribine is provided, saturated with cladribine.Following Table I has been listed the saturated rings dextrin solution for the given cyclodextrin of given cyclodextrin concentration.

Table I

With the cladribine in the Table I molar concentration of cyclodextrin is drawn, be expressed as Fig. 1.For cladribine-HP β CD, cladribine-HP β CD and 0.1% HPMC and line that cladribine-γ CD draws are represented maximum cladribine dissolving under the experiment condition, i.e. the HTA ratio of cladribine concentration and cyclodextrin concentration.Area representative above every figure line exists the situation of excessive insoluble cladribine.On behalf of cyclodextrin, the area below every figure line surpassed the situation that complex is maintained amount required in the solution.Obviously, the data among Table I and Fig. 1 show that HPMC (a kind of known compound accelerant) is not effect when lower concentration, and has negative effect when higher concentration.

The HTA figure of the cladribine that shows in Fig. 1-HP β CD is an approximately linear; This is indicating 1: 1 complex, and wherein the cyclodextrin of the medicine of 1 molecule and 1 molecule is compound.Fig. 1 also shows, needs extra cyclodextrin that cladribine is maintained in the complex.For example, in the situation of gamma-cyclodextrin, need about 0.10 mole of γ CD that about 0.01 mole of cladribine is maintained in its saturated complex; In the situation of HP β GD, need about 0.10 mole of cyclodextrin that about 0.017 mole of cladribine is maintained its saturated complex.But in the situation of γ CD, when the cyclodextrin of higher concentration, drug solubility significantly improves; For the γ CD of about 0.15 molar concentration, the slope of line changes, and shows 1: 2 complex that has formed cladribine and cyclodextrin, that is to say that the γ-CD of the cladribine of 1 molecule and 2 molecules is compound, and the latter surrounds and protect the cladribine molecule in fact.

Think that under high cyclodextrin concentration the γ-CD of 2 molecules is each other by hydrogen bonded, and the cladribine molecule is wrapped into cavity between them.Think that this is a process progressively, wherein at first form 1: 1 complex, the γ-CD in second γ-CD molecule and 1: 1 complex forms 1: 2 complex by hydrogen bonded then.Certainly, often obtained 1: 1 and the mixture of 1: 2 complex, but to preponderate be favourable to 1: 2 complex.Thereby under the situation of γ-CD, the cyclodextrin of about 0.20 molar concentration can maintain about 0.017 mole of cladribine in its saturated complex.Then, when higher cyclodextrin and drug level, for the amount of the required cyclodextrin of the cladribine of specified rate, difference is less between γ CD and HP β CD, and compares with HP β CD, and γ-CD can dissolve more cladribine pro rata.Because 1: 2 complex that forms under the γ of higher concentration CD is than 1: 1 stronger complex of complex, cladribine in the saturated solution that forms when 1: 2 such complex discharges medicine in the body fluid at mucosa place is more unsettled, promptly have the thermodynamic activity higher, help medicine and move by the stronger of mucosa than the cladribine that discharged the complex from 1: 1.With the complex of γ CD also be favourable because γ CD is natural cyclodextrin, thereby have the less relative toxicity of organizing.And, under the situation of solid oral dosage form, think and can protect cladribine to avoid the attack of gastric acid better with 1: 2 complex of γ-CD, surround drug molecule because it can be gone up substantially with cyclodextrin, thereby can be applicable to purpose of the present invention uniquely preferably.

The preparation of embodiment 2 cladribines-cyclodextrin complexes

A part:, make cladribine and HP β CD or γ CD compound by following conventional method.

With the concentrated solution of the waterborne suspension and the cyclodextrin of excessive cladribine (for gamma-cyclodextrin, about 27%; For HP β CD, about 40%) mixed about 9 hours in stirring at room.This can reach balance.Filter and remove the excessive not compound cladribine that may exist.In order to form the saturated cladribine-cyclodextrin complexes of solid, be integrated into that solid is sucked or oral tablet before, lyophilization aqueous cladribine-cyclodextrin solution.Freeze drying process comprises: make composite solution drop to-40 ℃ to about-80 ℃ about 2 to 4 hours of temperature maintenance, preferred about 3 to 4 hours freezing stage approximately rapidly, for example about-45 ℃ temperature maintenance is about 200 minutes, carry out subsequently at about-25 ℃ of drying stages first of keeping about 80-90 hour, typically under low pressure, carry out redrying stage of keeping about 15-20 hour then at about 30 ℃.

By HPLC (use 3 microns posts of Hypersil ODS and, carry out UV detection), can analyze product, to determine the cladribine in the end-product and the weight ratio of cyclodextrin by aforementioned conventional method preparation at 264nm based on the mobile phase of acetonitrile.Pass through methods known in the art, comprise for example by checking outward appearance, determine total impurities content by HPLC, use the KarlFischer titrator to determine water content, detect dissolubility by standard method, for example use USP＜711〉Apparatus II device and carry out UV detection at 264nm, check the content uniformity and analyze active component by HPLC and carry out quantitative assay, can further characterize the end-product goods.

The B part:

By aforesaid conventional method, be prepared as follows 2 batches of cladribine/cyclodextrin products: the FD02 and the FD03 that has used HP β CD that have used γ-CD:

For every batch, with purified water (for FD02,585ml; For FD03,575ml) be assigned in 1 liter of vial.Weighing gamma-cyclodextrin (116g) and 2-HP-(115g) added in the water that stirs lentamente through 30 minutes.The weighing cladribine (for FD02,2.53g; For FD03,2.76g), add in the cyclodextrin solution of stirring separately.Supersound process solution 20 minutes.The settled solution that obtains in stirring at room 9 hours.Solution is packed into (20ml solution/bottle) in the 100ml lyophilizing bottle, partly clog the bottle of filling.Lyophilizing comprises :-45 ℃ freezing about 3.3 hours, under-25 ℃, the pressure of 100 millitorrs, carried out first drying stage about 85.8 hours, carry out about 17.5 hours of redrying stage at 30 ℃, as described below:

Lyophilization cycle

(use 3 microns posts of Hypersil ODS and by HPLC based on the mobile phase of acetonitrile, carry out UV detection at 264nm), analyzed by the FD02 of preceding method preparation and cladribine/cyclodextrin of FD03 batch, found that empirically it has following characteristics:

Complex	Weight: weight	Weight ratio
Complex	Weight: weight	Weight ratio	Cladribine: γ-CD	2.53g∶116.0g	1∶46
Cladribine: HP β CD	2.76g∶115.0g	1∶42	Cladribine: γ-CD	2.53g∶116.0g	1∶46

By DSR and X-ray diffraction method, analyzed product, to determine any free cladribine in the freeze-drying prods.Importantly, sample does not show transition in 210 ℃ to 230 ℃ zone, and the latter is relevant with the fusion of crystal cladribine.In both cases, in 210 ℃ to 230 ℃ scope, it is active all not record significant heat, shows that the complex that obtains when lyophilizing finishes does not have the free crystal cladribine of any significant quantity, considers Sensitivity of Analytical Method (3% w/w at the most).Do not have the peak of crystal cladribine in the X-ray diffraction vestige of 2 reply compound FD02 and FD03, support this conclusion yet.

As mentioned above, these cladribines: cyclodextrin complexes has following cladribine: the cyclodextrin weight ratio: for cladribine: gamma-cyclodextrin is about 1: 46; For cladribine: HP-is about 1: 42.Cladribine near these values: the cyclodextrin weight ratio, for example about 1: 35 to about 1: 50 is optimal.These ratios can according to the concrete cyclodextrin that uses and in composite solution the amount and the combined temp of cyclodextrin change.

Embodiment 3

Pharmacokinetic study

In Bi Erge sleuth model, estimated cladribine with the bioavailability of γ CD or HP β CD compound tense.The data that expection obtains from this model are used for human body can be representative.

Saturated cladribine-cyclodextrin complexes the FD02 and the FD03 that will partly prepare as the B of embodiment 2 are used to prepare oral and buccal tablet.Composite materials and magnesium stearate are passed 18 orders (0.9mm) sieve, mix 5 minutes, use the compacting of 10mm stamping machine.The slightly protruding shape and the shape of lower plane hypotenuse above this 10mm tablet has.Factory formula is as follows:

Table II, the A part

* the complex of this amount contains the 5mg cladribine/sheet of having an appointment.

Table II, the B part

The character of finished tablet	Cladribine: γ-CD tablet RDT-0418/C	Cladribine: HP-beta-schardinger dextrin-tablet RDT-0418/D
The character of finished tablet	Cladribine: γ-CD tablet RDT-0418/C	Cladribine: HP-beta-schardinger dextrin-tablet RDT-0418/D	Average weight	237.0mg	217mg
Hardness	4.0Kp	6.72Kp	Average weight	237.0mg	217mg
Hardness	4.0Kp	6.72Kp	Fragility	0.5％	0.4％
Thickness	3.8mm	3.3mm	Fragility	0.5％	0.4％
Thickness	3.8mm	3.3mm	Disintegration time	8 minutes	8 minutes

Followingly in Bi Erge sleuth model, bioavailability and pharmacokinetic study have been carried out.

Make that (Dunakeszi, the male complete erg sleuth (being called PM01-PM06) of the outbreed that Hungary) obtains arbitrarily obtains laboratory diet and water from IDRI.In whole research, use identical Canis familiaris L. so that between object with object in the diversity minimum.Following bioavailability and the pharmacokinetic study of carrying out.

In the experimental stage first time, use 5mg cladribine (0.25mg/ml is in isotonic saline solution) for the experimental subject intravenous.In 48 hours, collect blood sample with different intervals.In the experimental stage second time, half object oral cavity accept aforesaid contain saturated cladribine-γ CD or-tablet of HP β CD complex.In 48 hours, collect serial blood sample.Experimental stage repeats the experimental stage for the second time for the third time, accepted the object of gamma-cyclodextrin buccal tablet just and accept the HP-buccal tablet now, the HP-tablet receiver of second stage accepts the gamma-cyclodextrin buccal tablet now.In the 4th and the 5th repeated experiments stage

experimental stage

2 and 3, just oral administration gives tablet.

By HPLC and LC/MS/MS method, detect the cladribine level in the blood.Use TopFit 2.0 pharmacokinetics and pharmacodynamics data analysis system to carry out the pharmacokinetic analysis of data.The result that will contrast the bioavailability study of (intravenous) and cladribine-cyclodextrin complexes is presented at Table III to VII, and is summarised in the Table VIII.

The header of each row in the Table III is defined as follows:

C _InitialIt is the extrapolated value of injecting when finishing;

C _FirstBe the concentration of behind application dosage, measuring first in 5 minutes;

T1/2 terminal is that the half-life is eliminated at the end eventually;

AUD is the area under the data that record, with linear trapezoidal rule integration;

AUD _ExtBe from the time point of last measurement to infinite extrapolation area;

AUC is extrapolated to infinite AUD;

CI _TotBe CLTB (dosage/AUC); With

MRT _TotIt is mean residence time.

Table IV to each header that is listed as among the VII is defined as follows:

C _MaxIt is the peak concentration that records;

T _MaxBe to C _MaxTime;

T1/2 terminal is that the half-life is eliminated at the end eventually;

AUC is extrapolated to infinite AUD;

MRT _TotIt is mean residence time; With

F is the bioavailability of expressing with %.

(PE SCIEX, Foster City US), determine peak area, calibration curve, accuracy, precision value and concentration to use Analyst Software 1.1.For calculating mean value and standard deviation, Excel 5.0 softwares have been used.The concentration ratio of operational analysis thing and internal standard product is with respect to their peak area ratio, the match calibration curve.By the linear regression analysis of the least square of weighting, fitting a straight line on experimental point.The weighting scheme that uses is 1/ concentration square (a pg/ml blood plasma).

Use the early stage time point in back at intravenous, plasma concentration has surpassed the upper limit of calibration curve.Thereby inject the more sample of a small amount of again, to confirm the linear detector reaction.Because standard substance/internal standard product can accept to surpass the concentration of 100ng/ml than keeping identical in suitable detector reaction.For calculating mean value, standard deviation and CV%, Excel 5.0 softwares have been used.Do not accept to be lower than the level of quantitative limit.The concentration calculating mean value and the standard deviation value that record after only intravenous being used.

Table VIII

*: get rid of (highly unusual) because the value that obtains is compared with other object from average

*: get rid of (because study subject has been swallowed pharmaceutical dosage form) from average

A: excessive cyclodextrin is to be used for 10 times of saturated complex approximately.

The total amount of the cyclodextrin in these researchs is to the about 2.5gm of 5mg cladribine.

Based on the curve of each plasma concentration, carry out pharmacokinetic analysis to the time.The meansigma methods and the standard deviation of the parameter that calculating obtains from each data.Use linearity-trapezoidal rule calculate from 0 to the plasma concentration-time graph of last measurement concentration area (AUD).Use the terminal point regression line, calculate in the following manner and be extrapolated to infinite area (AUC _{T → ∞}): AUC _{T → ∞}=C _CalC/ λ _z

Wherein, C _CalcThe plasma concentration of representative by estimating greater than the regression line of the last sampling time point of quantitative limit in the concentration that records, λ _zThe rate constant that representative calculates from the regression line.

The vision of the linear segment by semilog plot is determined, selects the each point of the decision regression line (being terminal stage).By the part area is added together, calculate area under the master curve (AUC): AUC=AUD+AUC _{T → ∞}Be used in the actual dose that moment uses and make the standardization of AUC value.In order to calculate bioavailability, will suck/oral AUC/ dose value is divided by intravenous AUC/ dose value.

Obtained individual blood plasma level-time graph.Use the back at intravenous and find the small individuals differences.After the very fast initial reduction, cladribine be about 10 hours through not eliminating the half-life.Confirmed that the average overall clearance rate is 17ml/min/kg.In sucking the process of using 2 kinds of preparations, cladribine: the breaking-in period of gamma-cyclodextrin complex is than cladribine: HP-beta-schardinger dextrin-complex is long.Although all shown higher interindividual variation sucking with Orally administered back peak concentration and absorb feature, Zong exposure value (AUC) shows much lower diversity.Confirmed that oral administration biaavailability is good: gamma-cyclodextrin and HP-complex are respectively 50 ± 3% and 45 ± 5%.It is lower to suck the bioavailability value: the gamma-cyclodextrin complex be 37 ± 10%, the HP-complex be 30 ± 4.5%.

The result who has shown the Canis familiaris L. pharmacokinetic study of further contrast gamma-cyclodextrin complex and mixture among the Table I X below." oral complex " row have been described the absolute bioavailability result of the 5mg cladribine in the 2.5g gamma-cyclodextrin; This is 10 times of cyclodextrin amount in the saturated complex of 5mg cladribine approximately.Said preparation is identical with " oral mixture " row, has just prepared complex, rather than has only mixed each component.The result of " oral mixture " and " oral complex " is comparable, shows the excessive cyclodextrin that use is big like this, can form complex in course of dissolution, disturbs dissolving, and bioavailability is produced negative interaction.The Canis familiaris L. and identical experimental technique identical as preceding use.These results show, use excessive cyclodextrin not reach re-set target, promptly be substantially devoid of and surpass patient's differences that the saturated cladribine-cyclodextrin complexes that will all basically cladribines maintains the cyclodextrin of minimum required in the complex provides enhanced bioavailability and reduction.

Table I X

* get rid of from meansigma methods

A: excessive cyclodextrin is to be used for 10 times of saturated complex approximately.The total amount of the cyclodextrin in these researchs is for the about 2.5gm of 5mg cladribine.

The result of aforementioned pharmacokinetic study is illustrated among Fig. 2-6.

Fig. 2 shown use the 5mg single dose with above-mentioned different preparations after, the curve of blood plasma of cladribine in Canis familiaris L., wherein data are meansigma methods ± standard deviations, every group of 5-6 animal.With the mean drug concentration (pg/ml) of blood plasma to the time (hour) draw.Although each experiment was carried out 48 hours, only preceding 6 hour meters are shown in the drawings; After 6 hours, most of concentration have been returned to or near baseline, have not therefore shown in the drawings.Think that intravenous value (◇) can produce 100% bioavailability, and contrasted oral and blood plasma level buccal forms with it." description of drawings " that the implication of sign sees above.Find that in these experiments to be not so good as oral formulations effective with the buccal lozenge of the saturated complex of gamma-cyclodextrin (*) and HP-(x), this can easily find out from Fig. 2.About the result of 5 kinds of different oral formulations, can more easily find out with reference to figure 3-5.

Fig. 3 has contrasted the curve of blood plasma of intravenous preparation (◆), oral saturated cladribine-gamma-cyclodextrin complex formulation (●) and oral saturated cladribine-HP-complex formulation (▲) (shown in Figure 2).These two kinds of oral formulations all provide ideal feature.

Fig. 4 has contrasted oral capsule (zero) cladribine-γ CD complex of physical mixture of oral saturated cladribine-gamma-cyclodextrin complex formulation (●), cladribine and 10 times of excessive gamma-cyclodextrins and oral capsule () curve of blood plasma shown in Figure 2 of 10 times of excessive gamma-cyclodextrins.Therefrom as can be seen, excessive cyclodextrin can reduce the amount of the cladribine in the blood plasma, particularly preceding 1 hour after using.

Fig. 5 has contrasted the curve of blood plasma of oral capsule (△) (shown in Figure 2) of the physical mixture of oral saturated HP-complex formulation (▲) and cladribine and 10 times of excessive HP-.Can be readily seen that once more that therefrom excessive cyclodextrin can reduce the amount of the cladribine in the complex.In this case, minimizing has been seen in preceding 2 hours after using.

Fig. 6 has illustrated for area (AUC) under the cumulative curve of 8 kinds of preparations shown in Figure 2 (pgxh/ml).Equally, data are meansigma methodss of every group of 5-6 animal.

Thereby each figure illustrates Table III to the shown content of the data among the IX, promptly make being substantially devoid of of Peroral solid dosage form or saturating mucosa dosage form and surpass the saturated cladribine-cyclodextrin complexes that will all basically cladribines maintains the cyclodextrin of minimum required in the complex and can strengthen bioavailability, have acceptable patient's differences simultaneously.

Think that the content of front only is used to explain principle of the present invention.And; because those skilled in the art can easily expect carrying out many improvement and variation; shown in should not limiting the invention to and accurate formation and operation that describe, therefore, all suitable improvement of its that can realize and equivalent are all in claimed scope of the present invention.

Claims

1. make the pharmaceutical composition of saturated cladribine-cyclodextrin complexes of containing of solid oral dosage form, described compositions is substantially devoid of and surpasses the cyclodextrin that will all basically cladribines maintains minimum required in the complex, wherein cyclodextrin is a gamma-cyclodextrin, HP-, hydroxypropyl-gamma-cyclodextrin, DM-, methylated at random beta-schardinger dextrin-, carboxymethyl-beta-cyclodextrin or sulfur butyl-beta-schardinger dextrin-.

2. according to the compositions of claim 1, wherein cyclodextrin is a gamma-cyclodextrin.

3. according to the compositions of claim 1, wherein cyclodextrin is a HP-.

4. according to the compositions of claim 2, wherein complex comprises 1: 2 cladribine: the gamma-cyclodextrin complex.

5. according to the compositions of claim 2 or 3, wherein the weight ratio of cladribine and cyclodextrin is 1: 35 to 1: 50.

6. according to the compositions of claim 2, wherein the weight ratio of cladribine and gamma-cyclodextrin is 1: 46.

7. according to the compositions of claim 3, wherein the weight ratio of cladribine and HP-is 1: 42.

8. according to each the compositions in the claim 1,2 and 3, wherein cladribine corresponding with the mol ratio of cyclodextrin be arranged in the point of cladribine on the phase solubility figure of the saturated complex of the cyclodextrin of variable concentrations.

9. compositions according to Claim 8, wherein cyclodextrin is a gamma-cyclodextrin, and this point is taken from and indicated the formation cladribine: the phase solubility figure part of 1: 2 complex of gamma-cyclodextrin.

10. saturated cladribine-cyclodextrin complexes is used for the application of administration with the solid oral dosage form of the reactive condition symptoms of treatment cladribine in preparation, described dosage form is substantially devoid of and surpasses the cyclodextrin that will all basically cladribines maintains minimum required in the complex, wherein the reactive disease of cladribine is selected from multiple sclerosis, rheumatoid arthritis, inflammatory bowel and leukemia, wherein cyclodextrin is a gamma-cyclodextrin, HP-, hydroxypropyl-gamma-cyclodextrin, DM-, methylated at random beta-schardinger dextrin-, carboxymethyl-beta-cyclodextrin or sulfur butyl-beta-schardinger dextrin-.

11. according to the application of claim 10, wherein the reactive disease of cladribine is selected from multiple sclerosis, rheumatoid arthritis and leukemia,

12. according to the application of claim 11, wherein the reactive disease of cladribine is a multiple sclerosis.

13. according to each the application in the claim 10,11 and 12, wherein cyclodextrin is a gamma-cyclodextrin.

14. according to each the application in the claim 10,11 and 12, wherein cyclodextrin is a HP-.

15. according to the application of claim 10, wherein the weight ratio of cladribine and cyclodextrin is 1: 35 to 1: 50.

16. according to the application of claim 13, wherein the weight ratio of cladribine and gamma-cyclodextrin is 1: 46.

17. according to the application of claim 14, wherein the weight ratio of cladribine and HP-is 1: 42.

18. according to the application of claim 13, wherein complex comprises 1: 2 cladribine: the gamma-cyclodextrin complex.

19. saturated cladribine-cyclodextrin complexes is used for improving the application of solid oral dosage form of the oral administration biaavailability of cladribine in preparation, described dosage form is substantially devoid of and surpasses the cyclodextrin that will all basically cladribines maintains minimum required in the complex, wherein cyclodextrin is a gamma-cyclodextrin, HP-, hydroxypropyl-gamma-cyclodextrin, DM-, methylated at random beta-schardinger dextrin-, carboxymethyl-beta-cyclodextrin or sulfur butyl-beta-schardinger dextrin-.

20. according to the application of claim 19, wherein cyclodextrin is a gamma-cyclodextrin.

21. according to the application of claim 19, wherein cyclodextrin is a HP-.

22. according to claim 19,20 or 21 each application, wherein the weight ratio of cladribine and cyclodextrin is 1: 35 to 1: 50.

23. according to the application of claim 20, wherein the weight ratio of cladribine and gamma-cyclodextrin is 1: 46.

24. according to the application of claim 21, wherein the weight ratio of cladribine and HP-is 1: 42.

25. according to the application of claim 20, wherein complex comprises 1: 2 cladribine: the gamma-cyclodextrin complex.

26.1: 2 cladribines: gamma-cyclodextrin complex.

27.1: 1 cladribine: gamma-cyclodextrin complex and 1: 2 cladribine: the mixture of gamma-cyclodextrin complex, wherein 1: 2 complex is main.