CN100408028C - Oral formulations of cladribine - Google Patents

Abstract

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

Description

The oral formulations of cladribine

Technical field

The present invention relates to make the compositions that comprises compound cladribine-cyclodextrin complexes of solid oral dosage form, also relate to the method for the oral administration biaavailability 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 exists with white, non-absorptive crystal powder form, is made up of independent crystal and crystal accumulation body.

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 respect to reduce cost relevant of parenteral administration cost with dosage forms for oral administration.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 transport realization by per os or saturating mucosa.

But up to now, there is following shortcoming in the oral delivery cladribine: 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 TherapieExper., 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 by 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.

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.

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, 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 amorphous cyclodextrin can combine with cladribine, formation can be integrated into the particularly advantageous product in the solid oral dosage form.This product is compound cladribine-cyclodextrin complexes, and the solid oral dosage form that contains it can improve oral administration biaavailability and/or realize between the lower patient of medicine and/or difference in the patient.

The invention provides compound cladribine-cyclodextrin complexes, it be (a) cladribine with the unbodied inclusion complex of amorphous cyclodextrin and (b) unbodied free cladribine combine the amorphous mixture closely of the non-inclusion complex of formation with amorphous cyclodextrin, with the pharmaceutical composition of making solid oral dosage form, it comprises described complex.Thereby cyclodextrin itself is unbodied, and the inclusion complex of itself and cladribine is unbodied (and preferably cladribine is saturated), and the free cladribine that forms non-inclusion complex is unbodied.

The present invention also provides the method that improves or strengthen the oral administration biaavailability of cladribine, comprise the pharmaceutical composition of making solid oral dosage form to the object dosage forms for oral administration of needs, it comprises compound cladribine-cyclodextrin complexes, the latter be (a) cladribine with the unbodied inclusion complex of amorphous cyclodextrin and (b) unbodied free cladribine combine the amorphous mixture closely of the non-inclusion complex that forms with amorphous cyclodextrin, this dosage form can make the amount of the cladribine in enclose and the non-inclusion complex maximize.

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 oral administration biaavailability of reactive disease of treatment cladribine and enhancing cladribine in preparation.

Further, the invention provides the method for the compound cladribine-cyclodextrin complexes of preparation, it comprises the steps:

(i) making cladribine and amorphous cyclodextrin is about 40 combinations to about 80 ℃ water in temperature, keeps described temperature about 6 to about 24 hours;

(ii) cool off resulting aqueous solution to room temperature; With

(iii) the refrigerative solution of lyophilizing obtains unbodied product.

On the other hand, the invention provides the pharmaceutical composition that can obtain by the method that comprises the steps:

(i) making cladribine and amorphous cyclodextrin is about 40 combinations to about 80 ℃ water in temperature, keeps described temperature about 6 to about 24 hours;

(ii) cool off resulting aqueous solution to room temperature;

(iii) the refrigerative solution of lyophilizing obtains unbodied product; With

(iv) unbodied product is mixed with solid oral dosage form.

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 unique figure is the result's of phase solubility research a diagram, the HP-(HP β CD) of wherein using different molar concentrations is to different cladribine molar concentration mappings, represents below the compound data point that obtains under the embodiment 2 described conditions with ().

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 " inclusion complex " refers to the compound of Cladribine and selected cyclodextrin, and wherein the hydrophobic part of Cladribine molecule (nitrogenous ring system) inserts in the hydrophobic cavity of cyclodextrin molecular. This is generally referred to as the cyclodextrin complexes of medicine.

Term " non-inclusion complex " refers to not be the compound of inclusion complex; Insert in the cyclodextrin cavity different from the hydrophobic part of Cladribine, non-inclusion complex mainly by the hydroxyl on the hydroxyl on " free " Cladribine (namely not the Cladribine in inclusion complex) and amino and the cyclodextrin ring outer surface (for example, in the situation of HP-β-CD, the hydroxypropyl on the glucose ring and hydroxyl) between Hydrogenbond form. This is the combination looser than inclusion complex.

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, compound Cladribine-cyclodextrin complexes of making solid oral dosage form is provided, its be (a) Cladribine with the unbodied inclusion complex of amorphous cyclodextrin and (b) unbodied free Cladribine be combined the closely amorphous mixture of non-inclusion complex of formation with amorphous cyclodextrin, thereby after using, can provide between the bioavilability of improvement and/or lower patient and/or difference in the patient. The present invention is necessary that following combination: the amorphous property of raw material cyclodextrin, water solubility level (about 5mg/ml that Cladribine shows, room temperature), with its ability of Hydrogenbond, it obtains under described specified conditions utilizing hereinafter, and the special amorphous mixture of the oral administration biaavailability that is specially adapted to optimize Cladribine can be provided.

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 in this article about, neighbouring, rough or about. " pact " and number 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.

As used herein, term " unbodied " refers to non-crystal solid. The cyclodextrin that this paper comprises itself is unbodied, because each in them all is comprised of separately multiple isomers, and the compound of they and Cladribine also is unbodied. And, can select compound condition (recombination time of high temperature and prolongation, as mentioned below), in order to form super-saturated Cladribine solution. When cooling, because the amorphous property of compound and cyclodextrin, some excessive free Cladribines can not precipitate, but are captured in the immixture with (preferred saturated) unbodied Cladribine-cyclodextrin inclusion complexes with amorphous form. This excessive Cladribine forms loose combination or non-inclusion complex by Hydrogenbond and cyclodextrin. Then, this can further increase the amount of the Cladribine in the product; Because it is unbodied; also because it be with the immixture of unbodied inclusion complex in, expect that therefore this extra Cladribine can obtain certain protection of avoiding the hydrochloric acid in gastric juice degraded (although it may be protected like that not as the Cladribine in the inclusion complex form).

When with amorphous cyclodextrin in the compound of Cladribine unite when using, term " saturated " refer to that this compound is that Cladribine is saturated, that is to say that this compound contains under the compound condition of using the Cladribine of maximum that can compound with the cyclodextrin of specified rate (by means of inclusion and non-inclusion complex). Phase solubility research can be used for providing this information, such as hereinafter in greater detail (following also described in more detail compound condition). Perhaps, can obtain saturated compound, by in the aqueous solution of selected cyclodextrin, adding Cladribine simply, until solution no longer dissolves Cladribine empirically; At last, the excessive Cladribine that removal may exist (by filtration or centrifugal), Solutions in Freeze-drying obtains dry saturated compound.

Statement " basically ", as in " there is no ", refer to the accurate Calculation amount 20% in, in preferred 10%, more preferably in 5%.

Term " otherness between the patient " is the difference of pointing between the patient of its drug administration. Term " otherness in the patient " refers to the difference that same patient experiences 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 the gastric mucosa of its absorption the time, wherein the cladribine solution that is in the cyclodextrin of hyperpyrexia mechanical state and 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, suppose by dissolving (for example, by with fluid for example body fluid contact), can form the saturated cladribine solution in part according to dry compositions of the present invention, wherein cladribine is in high heating power activity (HTA) state, thereby helps absorbing.Cladribine has quite low (although not being insignificant) inherent aqueous dissolubility; In fact it have certain water solublity.The dissociate free cladribine that forms of enclose from saturated aqueous solution and non-inclusion complex absorbs through gastric mucosa and shows more stable activity level.

In view of foregoing, obviously to produce the optimal drug compositions of solid oral 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 amorphous cyclodextrin, this ratio is called the HTA ratio in this article.

Determine the HTA ratio empirically, and it be accredited as the ratio of cladribine and amorphous cyclodextrin, its correspondence can with the cladribine of the compound maximum of cyclodextrin of specified rate.Use experience method (for example phase solubility research) determines to use the saturated concentration of the dissolved cladribine of amorphous 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 amorphous 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, cladribine 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 selecting 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 inclusion complexes that forms, for example, this inclusion 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 selected amorphous cyclodextrin (for example HP-(HP β CD) or hydroxypropyl-gamma-cyclodextrin) or cladribine to begin as fixed variable, to wherein adding excessive another kind, differentiating different dissolubility data point (indicating saturated cladribine-cyclodextrin complexes), and draw the line that obtains.Typically, find to promote in the aqueous solution of the amorphous cyclodextrin that contains concentration known, to add cladribine under the condition of complex formation by rule of thumb.Usually, under heating, for example keep the regular hour, for example at least 6-9 hour, carry out compound at about 45 ℃ to about 60 ℃; Think to reach 24 hours, can obtain better result by being heated to up to about 80 ℃, keeping.Remove excessive sedimentary cladribine then, detect cladribine concentration subsequently.This concentration is being represented the amount for the given dissolved cladribine of amorphous 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 concentration of the cladribine in the selected amorphous cyclodextrin that is dissolved in concentration known.Draw with data point then, demonstrate 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 amount of the cladribine of the cyclodextrin of concentration that is used to form solution.Can understand that cladribine is about 5mg/ml at the aqueous dissolubility of room temperature, can be higher when high temperature.Therefore, the data point correspondence the amount that is dissolved in the cladribine in aqueous HP β CD or other the amorphous cyclodextrin under selected condition; When lyophilizing afterwards, this solution generates compound cladribine-cyclodextrin complexes, its be (a) cladribine with the unbodied inclusion complex of amorphous cyclodextrin and (b) unbodied free cladribine combine the amorphous mixture closely of the non-inclusion complex of formation with amorphous cyclodextrin.If reach equilibrium condition in recombination process, unbodied cladribine-cyclodextrin complexes will be that cladribine is saturated.

Those skilled in the art can understand, by multiple alternative method, and the concentration in the time of can identifying the saturated complex that forms cladribine and amorphous cyclodextrin (and HTA than).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 (comparing the bioavailability of improvement and/or the coefficient of variation with traditional method) is accompanied by the inclusion complex of cladribine and cyclodextrin and the mixture of non-inclusion complex.

Use unbodied in essence cyclodextrin, for example HP-, hydroxypropyl-gamma-cyclodextrin, methylated cyclodextrin etc. at random, water-soluble a little chemical compound cladribine (can carry out hydrogen bonded by its free hydroxyl group and amino) can improve the dissolubility of cladribine in these cyclodextrin solutions.Not only have the water-soluble of raising, and improved the cladribine beyond the actual enclose composite and the hydrogen bonded of cyclodextrin.

Those skilled in the art can understand, the phase solubility figure of every kind of given initial concentration ratio is representing the once starting point of research, based on this, can change variable (concentration of reactants, temperature and time), to promote the combination of inclusion complex and non-inclusion complex, be beneficial to the combination that in end product, forms arbitrary type of higher or lower ratio.Except the ratio of cladribine and cyclodextrin, temperature and/or dilution factor that empirical discovery can promote that balance forms to complex have been analyzed then, to promote to form cladribine and the inclusion complex of cyclodextrin and the mixture of non-inclusion complex according to different proportion of the present invention.

Thereby, for example,, can in solution, hold more cladribine in theory by from rarer cyclodextrin (that is) beginning than being used for the bigger water volume of dissolubility picture analysis, their more complex that breaks away from are formed.After the evaporation, some dissolved cladribines can tend to combine with cyclodextrin in non-inclusion complex mode.By changing initial dilution factor, can make balance form migration to inclusion complex or non-inclusion complex.Similarly,, can increase the water solubility of cladribine, reduce the stability of inclusion complex simultaneously, thereby promote non-inclusion complex by improving combined temp.Thereby, by changing combined temp, can make balance form migration to inclusion complex or non-inclusion complex.At last, can change recombination time, be beneficial to form according to the inclusion complex of cladribine of the present invention and cyclodextrin and the mixture of non-inclusion complex.

As institute's illustration hereinafter, (use rarer cyclodextrin solution, higher temperature and longer recombination time, as mentioned above), can make the cladribine maximization in solid amorphous mixture by extra cladribine is entered in the solution.When solution cooled down, the extensive unbodied character of these cyclodextrin did not allow to surpass itself and the excessive cladribine crystallization of cyclodextrin formation inclusion complex; After freeze-dried/lyophilizing, can obtain the amorphous mixture of cladribine-cyclodextrin inclusion complexes (it is unbodied) and unbodied free cladribine, wherein unbodied free cladribine by hydrogen bond and not compound cyclodextrin (with in addition with compound cyclodextrin) loosely combines, that is to say non-inclusion complex.

Confirm as embodiment,, can make the dissolving maximization, thereby accomplish this point by elevated temperature before lyophilization (for example) and stirred for several hour (as many as 24 hours) to about 50 °-80 ℃.The w/w ratio that obtains is about 1: 14 and 1: 11.Apparent optimum weight/weight ratio under the condition of these examples is wherein higher, or about 1: 14 cladribine: cyclodextrin.If add too many excessive cladribine in complex media, crystallization can take place in some cladribines so, and it causes generating some crystal cladribines again in product; This unwanted excessive cladribine not with amorphous cyclodextrin hydrogen bonded, and can reduce weight ratio not in solution.Therefore, need to control to the amount that only is dissolved in the solution above the amount that can form the excessive cladribine of inclusion complex carefully.The ideal amorphous mixture of unbodied inclusion complex and unbodied free cladribine can be called " compound cladribine-cyclodextrin complexes ", it comprises the complex of enclose and non-enclose/hydrogen bonded.Inclusion complex is the complex that inserts the cladribine in the hydrophobic cavity of selected amorphous cyclodextrin, but not the complex of enclose/hydrogen bonded is the unbodied free cladribine with cyclodextrin loosely hydrogen bonded.Estimate that when obtain product the cladribine of about 2/3 (60-70%) can be in non-inclusion complex, 1/3 (30-40%) that is left is in inclusion complex, as (17% the HP β CD solution, 45-50 ℃ combined temp was kept about 9 hours) of illustration hereinafter; Percentage ratio by the cyclodextrin that improve to use and/or control temperature can easily obtain product, and wherein more a high proportion of amorphous mixture is the form of inclusion complex.Under the situation of representational amorphous cyclodextrin HP-(HP β CD), about 1: 10 to about 1: 16 cladribine: the cyclodextrin weight ratio is suitable for the condition of example; Expect identical for this ratio of hydroxypropyl-gamma-cyclodextrin under these conditions.By cladribine is dissolved into rapidly in the aqueous medium, can characterize the material that obtains.

Lyophilization is also referred to as lyophilizing, comprises 3 root 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 at this paper whole incorporated by reference and Xiaolin (Charlie) Tang that relies on and Michael J.Pikal at PharmaceuticalResearch, the 21st volume, the 2nd phase, in February, 2004, the described principle of 191-200 can further be optimized this method.

Above-mentioned method needs unbodied cyclodextrin, rather than has a cyclodextrin of relatively low water miscible parent crystal, for example α-, β-or gamma-cyclodextrin, 2,6-DM-etc., because these cyclodextrin allow therefore can not form ideal amorphous mixture than forming the excessive cladribine crystallization of inclusion complex.If cladribine be highly hydrophobic/lipophilic, then can not use this method because under these circumstances, medicine can not have inherent water solublity/hydrogen bonded ability, and the mixture of the uniqueness that this paper obtains can not be provided.But in fact, cladribine has the aqueous dissolubility of 5mg/ml in room temperature, thereby during the medicine of significant quantity can be soluble in the aqueous phase simply, particularly is being higher than the temperature of room temperature; In addition, as under the situation of HP β CD, for example, and 2 hydroxy functional groups in the Deoxyadenosine part by cladribine, some cladribines can combine with 2-hydroxypropyl and free glucose-OH group in the cyclodextrin by hydrogen bond.

Cyclodextrin in the scope of the invention be natural cyclodextrin α-, β-and the amorphous derivant of gamma-cyclodextrin, wherein, 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.Although do as a whole the address usually, unbodied cyclodextrin is actually the mixture of many different entities, because substituent group can be positioned on the different hydroxyls of basic cyclodextrin structure.This causes the amorphous property of these cyclodextrin again, and this is actually well-known.And these cyclodextrin 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 amorphous 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 by 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 by C _1-3Alkyl, hydroxyl-C _2-4Alkyl or carboxyl-C _1-2Alkyl, or more specifically replaced 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 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 Pharmaceutical Bulletin 28:1552-1558 (1980); Yakugyo Jiho No.6452 (28March 1983); Angew.Chem.Int.Ed.Engl.19:344-362 (1980); U.S. Patent number 3,459,731 and 4,535,152; European patent number EP 0149197A and EP 0197571A; Pct international patent publication number WO 90/12035; 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/or provide the document of preparation, purification and the analysis guide of cyclodextrin to comprise following: Jozsef Szejtli, the Cyclodextrin Technology of KluwerAcademic Publishers (1988) in Cyclodextrin in Pharmaceuticals 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 Inc lusioncomplexes 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., Int.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 0094157A.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 amorphous cyclodextrin of cladribine: 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; Methylated at random beta-schardinger dextrin-.2-HP-(HP β CD), 2-hydroxypropyl-gamma-cyclodextrin (HP γ CD), methylated at random beta-schardinger dextrin-, beta-schardinger dextrin-sulfur butyl ether, carboxymethyl-beta-cyclodextrin (CM β CD) and carboxymethyl-gamma-cyclodextrin (CM γ CD) are particularly advantageous, particularly HP-and hydroxypropyl-gamma-cyclodextrin.

Explain and example as this paper, can help under the condition that complex forms, in liquid environment, prepare the compositions of the amorphous mixture that is used for unbodied free cladribine of the present invention and unbodied preferred saturated cladribine-cyclodextrin inclusion complexes.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 aqueous solution of amorphous cyclodextrin, (for example separate undissolved cladribine, by filtration or centrifugal), lyophilizing or lyophilization saturated solution form solid amorphous mixture.

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 that cladribine, cyclodextrin or complex form and select excipient.

Randomly prepare dosage form together and (see Handbook of Pha rmaceuticalExcipients 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.Pharma ceutical 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 solid oral dosage form is by forming with the aforesaid unbodied free cladribine of (for example about 1% weight) suitable bonding or lubricant (for example magnesium stearate) compacting in a small amount and the amorphous mixture (being compound cladribine-cyclodextrin complexes) of unbodied cladribine-cyclodextrin complexes (preferred saturated).

In some cases, improve dissolubility or strengthen permeability,, can further promote oral administration to absorb for example by improving microenvironment by adding various excipient and additive.

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.

Compositions of the present invention is specially adapted to the therapy as the reactive disease of any cladribine 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 aforesaid compound cladribine-cyclodextrin complexes of effective dose, the i.e. amorphous mixture of unbodied saturated cladribine-amorphous cyclodextrin complexes of You Huaing and unbodied free cladribine (for example, effectively measuring) for treatment multiple sclerosis, rheumatoid arthritis or leukemia.

Term " treatment effective dose " or " effective dose " are used to represent the treatment with the dosage of the therapeutic outcome that can realize effectively pursuing.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 Opinionin 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 after dosage forms for oral administration, be understood that, even from the best bioavailability of peroral dosage form, can not the expectation meeting near use the bioavailability that the back obtains at intravenous, particularly at early time point.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.Think at present, for the treatment of multiple sclerosis, use once the 10mg cladribine in the compound cladribine-cyclodextrin complexes of the present invention 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 once 10mg cladribine in the compound cladribine-cyclodextrin complexes of the present invention in this solid dosage forms every day for the patient, every month 5-7 days, continue 6 totally 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 (Rebif

, Betaseron

/ Betaferon

, Avonex

), it is identical with the naturally occurring albumen of finding in human body; Glatiramer acetate (Copaxone

), i.e. the aminoacid GLAT with chain (polymer); Natalizumab (Antegren

), a kind of monoclonal antibody; Alemtuzumab (Campath-1H

), 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 (Cylert

) and 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 (Celebrex for example for example

), rofecoxib (Vioxx ), meloxicam (Mobic

), valdecoxib (Bextra

), lumiracoxib (Prexige

) and etoricoxib (Arcoxia

).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 the antirheumatic of disease modulability, and it comprises methotrexate, chloroquine, leflunomide (Arava

), 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 (Enrel

), a kind of tumor necrosis factor inhibitors, infliximab (Remicade

), it also is a kind of anti-TNF medicine, Antril (Synergen) (Kineret

), promptly a kind of optionally IL-1 blocker, and Humira

, promptly 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 dosage forms for oral administration approach of single dosage form 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 HP-(HP β CD) of variable concentrations, compound as carrying out as described in the following examples 2.Filter and remove unnecessary undissolved cladribine.Detect the amount of the cladribine in the composite solution, obtain data point.Use the HP β CD of different concentration known to repeat this method, up to obtaining several data points.Draw with these data points then, each data point is represented the amount of the cladribine in the water that can be dissolved in the cyclodextrin that contains specific concentrations.Point on the line that is generated by data point is represented the ratio of product.Those skilled in the art will appreciate that,, can obtain identical result if excessive cyclodextrin is joined in the cladribine solution of concentration known.

Molar concentration with cladribine that obtains and cyclodextrin is drawn.Representational phase solubility figure shows in the accompanying drawings.The drawing line of cladribine-HP β CD is being represented for the dissolving of the cladribine of test condition, i.e. the ratio of the concentration of cladribine and the concentration of cyclodextrin.Area more than every drawing line is being represented the situation of excessive insoluble cladribine; Area below every drawing line is being represented the excessive situation of cyclodextrin.

The figure of the cladribine of Xian Shiing-HP β CD is an approximately linear in the accompanying drawings; This is indicating 1: 1 complex, and wherein the cyclodextrin of the medicine of 1 molecule and 1 molecule is compound.Accompanying drawing also shows, needs extra cyclodextrin that cladribine is maintained in the complex.For example, under selected condition, need about 0.14 mole of HP β CD keep about 0.049 mole of cladribine dissolving, it provides unbodied, the preferred saturated cladribine-HP β CD inclusion complex and the amorphous mixture of unbodied free cladribine (as non-inclusion complex) the most at last.Below under the condition of embodiment 2, can expect in the product signal portion cladribine not in inclusion complex, but by hydrogen bonded as non-inclusion complex with in the loose immixture that remains on it of amorphous form.

Embodiment 2 preparations are used for the cladribine-cyclodextrin complexes of human experimentation

By following method, make cladribine and HP β CD compound.

In the 825mL distilled water, dissolving 172.5g HP-(forming about 17% solution) adds cladribine then, stirs the mixture about 9 hours at about 45 to about 50 ℃.Continue to stir other 6-9 hour in room temperature.Filter and remove all undissolved cladribines, make solution be cooled to room temperature.In order to form the amorphous mixture of unbodied cladribine-cyclodextrin complexes and unbodied free cladribine, before introducing the Peroral solid dosage form tablet, lyophilization aqueous cladribine-cyclodextrin solution.Freeze-drying process comprises: make composite solution drop to-40 ℃ of pacts rapidly and (for example extremely make an appointment with-80 ℃, the freezing stage of temperature maintenance-45 ℃ of pacts) about 2 to 4 hours (preferred about 3 to 4 hours), 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.

By aforesaid conventional method, be prepared as follows 2 batches of cladribines/cyclodextrin product, FD04 and FD05:

By immersing in the water-bath, the purified water (825mL) in 1 liter of glass jar is preheated to 48 ℃ (45 ℃-50 ℃ of target zones).The water of agitating heating is to produce in check central whirlpool.Weighing 2-HP-(172.50g) added in the water that heats lentamente through 40 minutes.The solution that obtains is continued to stir 10 minutes, guarantee that cyclodextrin dissolves fully.The weighing cladribine (for FD04,12.00g; For FD05,18.75g), add in the cyclodextrin solution that stirs, it becomes muddy, becomes clear then.The settled solution that obtains is maintained 48 ℃, continue to stir 9 hours.Continue to stir other 7 hours, make solution be cooled to room temperature simultaneously.

In the preparation of FD05, use more substantial cladribine, be a part of attempting optimizing this method; But, find that in this case the initial amount of cladribine is excessive, when finishing, the cooling step of batch FD05 observed precipitation.Filtering solution is to remove precipitation.The analysis showed that (measured value=87.2%) to the product that obtains under the situation of FD05, has the 16.35g cladribine to be integrated in the cyclodextrin complexes.For a batch FD04, do not need to filter, show that the amount of using is more suitable in the preparation of FD04, and, can optimize the FD05 method, thereby avoid filtration step by with more cladribine (16.35g, rather than the 18.75g) beginning of a small amount of.

After being cooled to room temperature, and under the situation of FD05, need to filter, solution is packed into (20ml solution/bottle) in the 100ml lyophilizing bottle, partly clog the bottle of filling, lyophilizing.Lyophilizing comprises :-45 ℃ freezing about 200 minutes, under-25 ℃, the pressure of 100 millitorrs, carried out first drying stage about 5,200 minutes, carry out about 1,080 minute of redrying stage at 30 ℃, as described below:

Table I

(use 3 microns posts of HypersilODS and by HPLC based on the mobile phase of acetonitrile, carry out UV detection at 264nm), analyzed by batch FD04 of preceding method preparation and cladribine/cyclodextrin product of FD05, it has following characteristics empirical discovery:

Table II

Lot number	Cladribine: HP β CD weight: weight	Cladribine: HP β CD weight ratio
			FD 04	12.00g∶172.50g	1∶14.38
FD 05	16.35g∶172.50g	1∶10.55

By DSR thermogram and X-ray diffraction method, analyzed product, to determine any free crystal 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 FD04 and FD05, support this conclusion yet.

Product is unbodied cladribine-HP β CD inclusion complex and the amorphous mixture that becomes the unbodied free cladribine of non-inclusion complex with the cyclodextrin hydrogen bonded.The cladribine that obtains: the weight ratio of HP β CD is about 1: 14 and 1: 11.

Generally speaking, amorphous mixture within the scope of the invention has about 1: 10 to 1: 16 cladribine: HP β CD weight ratio.

The preparation of embodiment 3 oral tablets

Use embodiment 2 described amorphous mixture FD04 and FD05 batch, produced tablet, be used for clinical research.

Use cladribine-2-HP β CD composite mix FD05, with 3,000 batch processes batch N0120; Use cladribine-HP β GD composite mix FD04, with 800 batch processes batch N0126.This main formula of two batches as shown in Table III.Batch N0120 is representing the 3.0mg tablet, and a batch N0126 is representing the 10mg tablet, is used for clinical research.

Table III

* be equivalent to 3.0mg cladribine/sheet

* is equivalent to 10.0mg cladribine/sheet

Following table has illustrated the method for production batch N0120 and N0126 tablet.

Table IV

1. with magnesium stearate and the approximately sorbitol powder premixing of equivalent;
	2. through 40 mesh sieves, with cladribine-HP β CD complex and the remaining sorbitol powder 1 liter of glass jar of packing into;
3. mixed each composition 10 minutes with 12rpm;
	4. through 40 mesh sieves, with premixed magnesium stearate/sorbitol powder glass jar of packing into;
5. mixed final mixture 5 minutes with 12rpm;
	6. respectively with the target pressing weight of 100mg and 200mg, be pressed into 3.0mg and 10.0mg tablet.

The 3.0mg tablet of batch N0120 and the 10.0mg tablet of batch N0126 all are circular, the flat hypotenuse of a side, and opposite side is slightly protruding.The 3.0mg tablet of batch N0120 has the average weight of 100mg, the thickness of 2.7mm, 0.2% fragility, the hardness of 4Kp and 3 minutes disintegration time.The 10.0mg tablet of batch N0126 has the average weight of 198mg, the thickness of 4.2mm, 1% fragility, the hardness of 2.8Kp and 5 minutes 42 seconds disintegration time.

The 3.0mg tablet of batch N0120 and the 10.0mg tablet of batch N0126 all are used for clinical research, are summarised in the following examples 5.

Embodiment 4

Clinical research: relative bioavailability

The purpose of this research is to estimate the relative bioavailability of 3 kinds of oral Cladribine formulations in MS (multiple sclerosis) patient: (1) is according to the preparation based on cyclodextrin of the present invention (tablet 1: complex FD05, promptly aforesaid lot number N0120 tablet); (2) preparation (tablet 2: contain the 3.0mg cladribine, 10mg carbopol 71G NF, 22.2mg dicalcium phosphate, 64.3mg lactose and 0.5mg magnesium stearate, lot number N0121) of adhesion mucosa; (3) (this capsule contains the 3.0mg cladribine to the hard gel capsule, 5.0mg carbopol 974P, 91.3mgAvicel PH101,100.0mg Avicel PH102,0.2mg silica sol and 0.5mg magnesium stearate, lot number RD03030), with once fixed subcutaneous administration cladribine (with reference to agent) contrast.

This research is 2 centers, open-label, at random, the 4 single dose researchs to intersection, has used 12 MS patients.The patient accepts 3 kinds of different fixed per os dosage (3.0mg) and fixed subcutaneous dosage 3.0mg at random.4 treatments day are separated at interval by at least 5 days no medicine.At each treatment stage, collected blood sample through 24 hours, be used to estimate the blood plasma cladribine.

By the HPLC/MS/MS method, measure the plasma concentration of cladribine.Using this method, find that the pass between concentration and the peak area ratio ties up to 100pg/ml to 50, is linear in the scope of 000pg/ml cladribine.The amount limit is 100pg/ml.Sample analysis carries out 16 operations.Need not to get rid of correction factor from the match of calibration curve, the accuracy of every special quality control sample can both satisfy the GLP requirement.

Analyze 576 parts of clinical plasma samples, and detected the concentration value of cladribine.The arrangement result, and be summarised in the following table (Table V and VI).In these tables, the definition below adopting: T _MaxIt is the time that in blood plasma, reaches Cmax; T _1/2It is the half-life of cladribine in blood plasma; C _MAXBe the Cmax of cladribine in blood plasma; AUC _InfBe under the data and curves that records, to be extrapolated to infinite area from 0; AUC _tIt is area under a curve (from 0 to last time point) in the data that record; Geom Mean is a geometric mean; CV is the coefficient of variation (relative standard deviation); LL is a lower bound; UL is the upper limit.

Table VI

Per os and the ratio of subcutaneous pharmacokinetic parameter and corresponding 2 sides, 90% confidence intervals (n=12) of cladribine research

* ratio that obtains by inverse conversion logarithm translation data (dosage standardization) and corresponding 95%LL.

Embodiment 5

Clinical research: dose response and absolute bioavailability

The purpose of this research is to estimate with the whole body utilization rate in MS (multiple sclerosis) patient behind two kinds of different fixedly oral dose dosage forms for oral administration cladribines, use (with reference to agent) contrast with fixed intravenous once, and estimate safety and the toleration of cladribine in this crowd.

This research is 3 centers, open-label, at random, the 3 single dose researchs to intersection, has used 26 MS patients.The patient accepts 2 kinds of different fixed oral doses (3.0mg and 10.0mg) and fixed intravenous dosages 3.0mg (inculcate use as 1 hour) at random.3 treatments day are separated at interval by at least 5 days no medicine.At each treatment stage, collected blood sample through 24 hours, be used to estimate the blood plasma cladribine.

By the HPLC/MS/MS method, measure the plasma concentration of cladribine.Using this method, find that the pass between concentration and the peak area ratio ties up to 100pg/ml to 50, is linear in the scope of 000pg/ml cladribine.The amount limit is 100pg/ml.Sample analysis carries out 16 operations.Except operation for the first time (because equipment fault, must eliminating it), all other operation all can be accepted.Need not to get rid of correction factor from the match of calibration curve, the accuracy of every special quality control sample can both satisfy the GLP requirement.

Analyze 858 parts of clinical plasma samples, and detected the concentration value of cladribine.The arrangement result, and be summarised in the following table (Table VII is to X).In these tables, the definition below adopting: T _MaxIt is the time that in blood plasma, reaches Cmax; T _1/2It is the half-life of cladribine in blood plasma; C _MAXBe the Cmax of cladribine in blood plasma; AUC _InfBe under the data and curves that records, to be extrapolated to infinite area from 0; AUC _tIt is area under a curve (from 0 to last time point) in the data that record; Geom Mean is a geometric mean; CV is the coefficient of variation (relative standard deviation); LL is a lower bound; UL is the upper limit; σ ²It is average variance; σ _B ²It is the average variance between the object; σ _W ²It is the average variance in the object; CV _TIt is total coefficient of variation; And CV _WIt is the coefficient of variation in the object.

Table VII

The summary statistical table (n=26) of cutting apart the pharmacokinetic parameter of analyzing the cladribine research that obtains by nothing

^*For T _MaxAnd T _1/2, the CV=SD/ average is for C _Max, AUC _InfAnd AUC _tBe the CV% geometric mean.

Table VIII

Per os and the ratio intravenous pharmacokinetic parameter of cladribine research and the lower bound (LL) of corresponding one-sided 95% confidence interval are (n=26)

* ratio that obtains by inverse conversion logarithm translation data (dosage standardization) and corresponding 95%LL.

Table I X

The ratio and corresponding 2 sides, 90% confidence intervals (n=26) of cladribine research

* ratio that obtains by inverse conversion logarithm translation data (dosage standardization) and corresponding 90%CI.

The component of variance (n=26) of Table X cladribine research

The source of variance	C max	AUC inf	AUC t
				Between (σ B 2)	.0380	.0487	.0492
Inner (σ W 2)	.1315	.0330	.0357
				Total (σ B 2+σ W 2)	.1695	.0816	.0849
CV T(％)	43.0	29.2	29.8
				CV W(％)	37.5	18.3	19.1

Wherein the PK parameter is that dosage is regulated, and $\mathrm{CV}=\sqrt{\exp \left({\mathrm{\σ}}^2\right)-1}$

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 (76)

1. make the pharmaceutical composition of solid oral dosage form, comprise compound cladribine-cyclodextrin complexes, its be (a) cladribine with the unbodied inclusion complex of amorphous cyclodextrin and (b) unbodied free cladribine combine the amorphous mixture closely of the non-inclusion complex of formation with amorphous cyclodextrin, wherein amorphous cyclodextrin is a HP-, hydroxypropyl-gamma-cyclodextrin, methylated at random beta-schardinger dextrin-, carboxymethyl-beta-cyclodextrin or sulfur butyl-beta-schardinger dextrin-.

2. according to the pharmaceutical composition of claim 1, wherein complex is that cladribine is saturated.

3. according to the compositions of claim 1 or 2, wherein amorphous cyclodextrin is a HP-.

4. according to the compositions of claim 1 or 2, wherein amorphous cyclodextrin is hydroxypropyl-gamma-cyclodextrin.

5. according to the compositions of claim 1 or 2, wherein the weight ratio of cladribine and amorphous cyclodextrin is 1: 10 to 1: 16.

6. according to the compositions of claim 5, wherein amorphous cyclodextrin is a HP-.

7. according to the compositions of claim 6, wherein the weight ratio of cladribine and HP-is 1: 14.

8. according to the compositions of claim 6, wherein the weight ratio of cladribine and HP-is 1: 11.

9. according to the compositions of claim 5, wherein amorphous cyclodextrin is hydroxypropyl-gamma-cyclodextrin.

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

11. according to the compositions of claim 1 or 2, wherein the cladribine of 30 to 40 weight % is in inclusion complex (a), and the cladribine of 70 to 60 weight % is in non-inclusion complex (b).

12. according to the compositions of claim 10, wherein the cladribine of 30 to 40 weight % is in inclusion complex (a), and the cladribine of 70 to 60 weight % is in non-inclusion complex (b).

13. compound cladribine-cyclodextrin complexes is administered for application in the solid oral dosage form of the reactive condition symptoms of treatment cladribine in preparation, described compound cladribine-cyclodextrin complexes be (a) cladribine with the unbodied inclusion complex of amorphous cyclodextrin and (b) unbodied free cladribine combine the amorphous mixture closely of the non-inclusion complex of formation with amorphous cyclodextrin, wherein the reactive disease of cladribine is selected from multiple sclerosis, rheumatoid arthritis, inflammatory bowel and leukemia, wherein amorphous cyclodextrin is a HP-, hydroxypropyl-gamma-cyclodextrin, methylated at random beta-schardinger dextrin-, carboxymethyl-beta-cyclodextrin or sulfur butyl-beta-schardinger dextrin-.

14. according to the application of claim 13, wherein complex is that cladribine is saturated.

15. according to the application of claim 13 or 14, wherein the reactive disease of cladribine is selected from multiple sclerosis, rheumatoid arthritis and leukemia.

16. according to the application of claim 15, wherein the reactive disease of cladribine is a multiple sclerosis.

17. according to the application of claim 13 or 14, wherein the weight ratio of cladribine and amorphous cyclodextrin is 1: 10 to 1: 16.

18. according to the application of claim 17, wherein amorphous cyclodextrin is a HP-.

19. according to the application of claim 18, wherein the weight ratio of cladribine and HP-is 1: 14.

20. according to the application of claim 18, wherein the weight ratio of cladribine and HP-is 1: 11.

21. according to the application of claim 17, wherein amorphous cyclodextrin is hydroxypropyl-gamma-cyclodextrin.

22. according to the application of claim 13 or 14, wherein the cladribine of 30 to 40 weight % is in inclusion complex (a), and the cladribine of 70 to 60 weight % is in non-inclusion complex (b).

23. compound cladribine-cyclodextrin complexes is used for improving the application of solid oral dosage form of the oral administration biaavailability of cladribine in preparation, described compound cladribine-cyclodextrin complexes be (a) cladribine with the unbodied inclusion complex of amorphous cyclodextrin and (b) unbodied free cladribine combine the amorphous mixture closely of the non-inclusion complex of formation with amorphous cyclodextrin, wherein amorphous cyclodextrin is a HP-, hydroxypropyl-gamma-cyclodextrin, methylated at random beta-schardinger dextrin-, carboxymethyl-beta-cyclodextrin or sulfur butyl-beta-schardinger dextrin-.

24. according to the application of claim 23, wherein complex is that cladribine is saturated.

25. according to the application of claim 23 or 24, wherein the weight ratio of cladribine and amorphous cyclodextrin is 1: 10 to 1: 16.

26. according to the application of claim 25, wherein amorphous cyclodextrin is a HP-.

27. according to the application of claim 26, wherein the weight ratio of cladribine and HP-is 1: 14.

28. according to the application of claim 26, wherein the weight ratio of cladribine and HP-is 1: 11.

29. according to the application of claim 25, wherein amorphous cyclodextrin is hydroxypropyl-gamma-cyclodextrin.

30. according to the application of claim 23 or 24, wherein the cladribine of 30 to 40 weight % is in inclusion complex (a), and the cladribine of 70 to 60 weight % is in non-inclusion complex (b).

31. compound cladribine-cyclodextrin complexes, its be (a) cladribine with the unbodied inclusion complex of amorphous cyclodextrin and (b) unbodied free cladribine combine the amorphous mixture closely of the non-inclusion complex of formation with amorphous cyclodextrin, wherein amorphous cyclodextrin is a HP-, hydroxypropyl-gamma-cyclodextrin, methylated at random beta-schardinger dextrin-, carboxymethyl-beta-cyclodextrin or sulfur butyl-beta-schardinger dextrin-.

32. according to the complex of claim 31, it is that cladribine is saturated.

33. according to the complex of claim 31 or 32, wherein amorphous cyclodextrin is a HP-.

34. according to the complex of claim 31 or 32, wherein amorphous cyclodextrin is the hydroxypropyl gamma-cyclodextrin.

35. according to the complex of claim 31 or 32, wherein the weight ratio of cladribine and amorphous cyclodextrin is 1: 10 to 1: 16.

36. according to the complex of claim 35, wherein amorphous cyclodextrin is a HP-.

37. according to the complex of claim 36, wherein the weight ratio of cladribine and HP-is 1: 14.

38. according to the complex of claim 36, wherein the weight ratio of cladribine and HP-is 1: 11.

39. according to the complex of claim 35, wherein amorphous cyclodextrin is the hydroxypropyl gamma-cyclodextrin.

40. according to the complex of claim 31 or 32, wherein the cladribine of 30 to 40 weight % is in inclusion complex (a), and the cladribine of 70 to 60 weight % is in non-inclusion complex (b).

41. the method for the compound cladribine-cyclodextrin complexes of preparation claim 31, it comprises the steps:

(i) cladribine and amorphous cyclodextrin are merged in temperature is 40 to 80 ℃ water, kept described temperature 6 to 24 hours;

(ii) cool off resulting aqueous solution to room temperature; With

The (iii) refrigerative solution of lyophilizing, obtain unbodied product, its be (a) cladribine with the unbodied inclusion complex of described amorphous cyclodextrin and (b) unbodied free cladribine combine the amorphous mixture closely of the non-inclusion complex of formation with described amorphous cyclodextrin.

42., also be included in the filtration step of step after (ii) according to the method for claim 41.

43. according to the method for claim 41 or 42, wherein step (i) is to carry out 45 to 60 ℃ temperature.

44. according to the method for claim 43, wherein step (i) is to carry out 45 to 50 ℃ temperature.

45. according to the method for claim 43, wherein step (i) is under agitation to carry out.

46. according to the method for claim 45, wherein step (i) was carried out 6 to 9 hours.

47. according to the method for claim 41 or 42, wherein step was (ii) carried out 6 to 9 hours.

48. according to the method for claim 43, wherein step was (ii) carried out 6 to 9 hours.

49. according to the method for claim 41 or 42, wherein step (iii) comprises the initial freezing stage, wherein solution is cooled to-40 to-80 ℃, and keeps 2 to 4 hours in described temperature.

50. according to the method for claim 48, wherein step (iii) comprises the initial freezing stage, wherein solution is cooled to-40 to-80 ℃, and keeps 2 to 4 hours in described temperature.

51., wherein,, solution is cooled to-45 ℃ in the step initial freezing stage (iii) according to the method for claim 49.

52., wherein the cladribine of 12.00 weight portions and the HP-of 172.50 weight portions are introduced in step (i) according to the method for claim 41 or 42.

53., wherein the cladribine of 16.35 weight portions and the HP-of 172.50 weight portions are introduced in step (i) according to the method for claim 41 or 42.

54., wherein the water of 825 parts by volume is introduced in step (i) according to the method for claim 52.

55., wherein the water of 825 parts by volume is introduced in step (i) according to the method for claim 53.

56. according to the method for claim 41 or 42, wherein step of freeze drying (iii) comprises:

(a) the initial freezing stage, wherein composite solution is cooled to-40 ℃ to-80 ℃ lasting 2-4 hour;

(b) drying stage first carried out 80-90 hour at-25 ℃; With

(c) the redrying stage, carried out 15-20 hour at 30 ℃.

57. according to the method for claim 56, the wherein freeze dried stage (a) is to carry out 3-4 hour at-45 ℃.

58. according to the method for claim 56, the wherein freeze dried stage (b) is to carry out under the pressure of 100 millitorrs.

59. the method for the pharmaceutical composition of preparation claim 1, it comprises the steps:

(i) cladribine and amorphous cyclodextrin are merged in temperature is 40 to 80 ℃ water, kept described temperature 6 to 24 hours;

(ii) cool off resulting aqueous solution to room temperature;

The (iii) refrigerative solution of lyophilizing, obtain unbodied product, its be (a) cladribine with the unbodied inclusion complex of described amorphous cyclodextrin and (b) unbodied free cladribine combine the amorphous mixture closely of the non-inclusion complex of formation with described amorphous cyclodextrin; With

(iv) unbodied product is mixed with solid oral dosage form.

60. according to the method for claim 59, wherein this method also be included in step (i) or (ii) after filtration step.

61. according to the method for claim 59 or 60, wherein the step of this method (i) is to carry out at 45 to 60 ℃.

62. according to the method for claim 61, wherein the step of this method (i) is to carry out at 45 to 50 ℃.

63. according to the method for claim 61, wherein the step of this method (i) is under agitation to carry out.

64. according to the method for claim 63, wherein the step of this method (i) was carried out 6 to 9 hours.

65. according to the method for claim 59 or 60, wherein the step of this method was (ii) carried out 6 to 9 hours.

66. according to the method for claim 59 or 60, wherein step (iii) comprises the initial freezing stage, wherein solution is cooled to-40 to-80 ℃, and keeps 2 to 4 hours in described temperature.

67., wherein,, solution is cooled to-45 ℃ in the step initial freezing stage (iii) according to the method for claim 66.

68., wherein the cladribine of 12.00 weight portions and the HP-of 172.50 weight portions are introduced in the step (i) of this method according to the method for claim 59 or 60.

69., wherein the cladribine of 16.35 weight portions and the HP-of 172.50 weight portions are introduced in the step (i) of this method according to the method for claim 59 or 60.

70., wherein the water of 825 parts by volume is introduced in the step (i) of this method according to the method for claim 68.

71., wherein the water of 825 parts by volume is introduced in the step (i) of this method according to the method for claim 69.

72. according to the method for claim 59 or 60, wherein the step of freeze drying of this method (iii) comprises:

(a) the initial freezing stage, wherein composite solution is cooled to-40 ℃ to-80 ℃ lasting 2-4 hour;

(b) drying stage first carried out 80-90 hour at-25 ℃; With

(c) the redrying stage, carried out 15-20 hour at 30 ℃.

73. according to the method for claim 72, the wherein freeze dried stage (a) is to carry out 3-4 hour at-45 ℃.

74. according to the method for claim 72, the wherein freeze dried stage (b) is to carry out under the pressure of 100 millitorrs.

75. according to the method for claim 59 or 60, wherein the preparation steps of this method (iv) comprises complex is mixed with magnesium stearate, and is pressed into tablet.

76. according to the pharmaceutical composition of claim 75, wherein with before complex mixes, with magnesium stearate and sorbitol powder premixing.