Stable nanoparticulate drug suspension

Stable nanoparticulate drug suspension

This application claims priority to U.S. Provisional Application No. 61/218,281, filed on June 18, 2009, the entirety of which is incorporated herein by reference.

field of invention

The present invention relates to a liquid suspension formulation comprising a low-solubility particulate drug compound and a method for preparing the formulation. The present invention is particularly applicable to a class of apoptosis-promoting compounds targeting Bcl-2 family proteins and thus the present invention further relates to a method of using a liquid suspension formulation for the treatment of a disease characterized by overexpression of said proteins will be.

Avoidance of apoptosis is a hallmark of cancer [Hanahan &amp; Weinberg (2000)<u>Cell</u> 100:57-70]. Cancer cells have to overcome the continual impact of cellular stresses such as DNA damage, oncogene activation, abnormal cell cycle progression, and the harsh microenvironment that causes normal cells to apoptosis. One of the main means by which cancer cells evade apoptosis is by upregulating anti-apoptotic proteins of the Bcl-2 family.

Compounds occupying the BH3 binding groove of the Bcl-2 protein are described, for example, in Bruncko <i>et</i><i></i><i>al</i><i>.</i> (2007) <u>J. </u><u>Med</u><u>. </u><u>Chem</u><u>.</u> 50:641-662]. The compound includes N-(4-(4-((4'-chloro-(1,1'-biphenyl)-2-yl)methyl)piperazine having the formula: Also known as ABT-737 -1-yl)benzoyl)-4-(((1R)-3-(dimethylamino)-1-((phenylsulfanyl)methyl)propyl)amino)-3-nitrobenzene-sulfonamide.

<img file="KR20120052937A_D0001.tif" />

ABT-737 is a Bcl-2 family protein (specifically, Bcl-2, Bcl-X<i>_L</i> and Bcl-w) with high affinity (&lt;1 nM). It exhibits single-drug activity against small cell lung cancer (SCLC) and lymphocytic malignancies, and potentiates the pro-apoptotic effect of other chemotherapeutic agents. ABT-737 and related compounds and methods for preparing these compounds are described in US Patent Application Publication No. 2007/0072860 (Bruncko et al.).

More recently, a series of compounds with high binding affinity for Bcl-2 family proteins have been further identified. These compounds and methods for their preparation are described in US Patent Application Publication No. 2007/0027135 (Bruncko et al.) (herein "'135 Publication"), which is incorporated herein by reference in its entirety, and has the formula It can be seen that it is structurally related to ABT-737.

The '135 publication states that previously known inhibitors of Bcl-2 family proteins may have either potent cellular potency or high systemic exposure following oral administration, but not both of these properties. A common measure of cellular potency of a compound is the concentration that elicits a 50% cellular effect (EC₅₀)am. A common measure of systemic exposure following oral administration of a compound is the area under the curve (AUC) obtained by graphing the plasma concentration of the compound with the time elapsed after oral administration. The previously known compounds mentioned in the '135 publication have low AUC/EC₅₀ ratio, which means that they are not effective orally. On the other hand, the compound of the above formula has a much higher AUC/EC than that of the conventionally known compounds.₅₀ It is described as having improved properties with respect to cellular potency and systemic exposure following oral administration.

One compound identified in "Example 1" of the '135 publication is N-(4-(4-((2-(4-chlorophenyl)-5,5-dimethyl, also known as ABT-263) -1-cyclohex-1-en-1-yl)methyl)piperazin-1-yl)benzoyl)-4-(((1R)-3-(morpholin-4-yl)-1-((phenyl) sulfanyl)methyl)propyl)amino-3-((trifluoromethyl)sulfonyl)benzenesulfonamide. This compound has a molecular weight of 974.6 g/mol and has the formula:

<img file="KR20120052937A_D0002.tif" />

ABT-263 is Bcl-2 and Bcl-X_LIt is considered that it binds with high affinity (&lt;1 nM) to Bcl-w and has similarly high affinity for Bcl-w. its AUC/EC₅₀ The ratio is reported as 56 in the '135 Gazette, which is more than one order of magnitude greater than that reported for ABT-737 (4.5). For the determination of AUC according to the '135 publication, each compound was administered 5 by oral gavage as a 2 mg/ml solution in a vehicle of 10% DMSO (dimethyl sulfoxide)/PEG-400 (polyethylene glycol with an average molecular weight of about 400). Administered to rats as a single dose of mg/kg.

Oral bioavailability (e.g., AUC after oral administration expressed as a percentage of AUC after intravenous administration) has not been reported in the '135 publication, but it follows from this that that of ABT-263 is substantially greater than that of ABT-737. It can be concluded that

Recently, the literature [Tse <i>et</i><i></i><i>al</i><i>.(2008) </i><u>Cancer</u><u></u><u>Res</u><u>.</u> 68(9):3421-3428] when administered to dogs, the oral bioavailability of a solution of ABT-263 in PEG400/DMSO was 22.4% and 60% Phosal^TM Supplementary data are available that the oral bioavailability of a solution of ABT-263 in PG (phosphatidylcholine + propylene glycol) and 30% PEG-400 and 10% ethanol was 47.6%.

Oxidation reactions represent an important degradation pathway for pharmaceuticals, especially when formulated in solution. Oxidation can occur by a number of pathways, including non-catalytic auto-oxidation of substrates by molecular oxygen, initiation of photolysis, hemolytic thermal cleavage, and metal catalysis. Various functional groups exhibit specific sensitivities to oxidation. In particular, thioethers can be cleaved either by hydrogen desorption at the α-position of the sulfur atom or by direct addition of an α-peroxyl radical or through a single electron transfer process, which transforms sulfides into sulfines, sulfones or sulfoxides. [See: Hovorka &amp; Schoneich (2001)<u>J. </u><u>Pharm</u><u>. </u><u>Sci</u><u>.</u> 90:253-269)

The (phenylsulfanyl)methyl group possessed by the compounds described in the '135 publication including ABT-263 is shown to have a thioether linkage, which is, for example, with oxygen or superoxide, hydrogen peroxide or a hydroxyl radical It appears to be prone to oxidation in the presence of the same reactive oxygen species. The '135 publication includes antioxidants among an extensive list of excipients useful for administration of the disclosed compounds.

However, it would be advantageous for pharmaceutical compositions to be less susceptible to oxidation of the active ingredient. In addition, the compositions are described in the '135 publication or Tse<i></i><i>et</i><i></i><i>al</i><i>. </i>(2008), <i>supra</i>It would be advantageous to be able to load the active ingredient higher than the solution composition of ].

The very low aqueous solubility '135 publication compounds, including ABT-263, pose problems associated with formulators, particularly when there is a need to maintain acceptable oral bioavailability, which is strongly dependent on solubility in aqueous media of the gastrointestinal tract. . Particle size reduction is often attempted as a method to increase the bioavailability of poorly water-soluble drugs, but it is often difficult and difficult to achieve a bioavailability comparable to that obtainable as a drug in solution form using solid particles of any size. , which can be considered as the ultimate challenge in particle size reduction.

Another problem with formulators providing suspensions of poorly water-soluble drug particles in a liquid medium is that suspended particles, especially very small particles of 1 μm in size or less, can lose particle size over time, e.g. through particle agglomeration. tends to grow Such an increase in particle size may make the suspension unstable and/or reduce its bioavailability. Surface modifiers such as surfactants are widely used, but not always successful. U.S. Patent No. 7,459,283 (Wertz &amp; Ryde) states that compositions contain a nanoparticulate active agent with lysozyme as a surface stabilizer.

See Moschwitzer et al. (2004)<u></u><u>Eur</u><u>. J.</u><u>Pharmaceut</u><u>. </u><u>Biopharmaceut</u>. 58:615-619], defined as a nanosuspension (nanocrystal (&lt;1000 nm diameter) dispersion in liquid phase) of omeprazole by dispersion of the drug in an aqueous medium containing 8.4% sodium bicarbonate and 1% poloxamer 188. ) is described. Physical stability studies have revealed an increase in median particle size after 3 days at 0°C; The authors concluded that the size increases. "Of course, I point out that these nanosuspensions do not have a long-term stability of two years." It is known that the chemical stability of omeprazole is greatly increased by formulating 50 or 100 mg/ml nanosuspension versus 5 mg/ml aqueous solution; The authors cite the inclusion of the crystal structure of the nanoparticles as a possible explanation for such stability.

In this regard, ABT-263, when prepared according to the '135 publication, is an amorphous solid; That is, it appears to be a poor candidate for nanosuspension formulations as it lacks crystallinity such as omeprazole.

A particular type of disease in need of improved therapy is non-Hodgkin's lymphoma (NHL). NHL is the sixth most prevalent new cancer in the United States, and it mainly occurs among patients in their 60s and 70s. NHL is not a single disease, but a family of related diseases classified based on several characteristics, including clinical attributes and histology.

One classification method is to divide the different histological subtypes into two main categories based on their intrinsic history: indolent and aggressive. In general, delayed subtypes progress slowly and are generally incurable, whereas aggressive subtypes progress rapidly and are potentially curable. The most common delayed subtype is follicular lymphoma, and diffuse giant cell lymphoma is the most common aggressive subtype. The oncoprotein Bcl-2 was first described in non-Hodgkin's B-cell lymphoma.

Treatment of follicular lymphoma usually consists of bio-based or combination chemotherapy. Combination therapy of rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP) is commonly used, as is the combination therapy of rituximab, cyclophosphamide, vincristine and prednisone (RCVP). Single-drug regimens of rituximab (which targets CD20, a phosphoprotein uniformly expressed on the surface of B-cells) or fludarabine are also used. The addition of rituximab to a chemotherapeutic regimen may provide improved response rates and increased progression-free survival.

Radioimmunotherapy, high-dose chemotherapy, and stem cell transplantation may be used to treat refractory or relapsed non-Hodgkin's lymphoma. Currently, there is no approved therapeutic regimen that provides cure, and current guidelines recommend treating patients in the context of a clinical trial, even in a first-line setting.

First-line treatment of patients with aggressive large B-cell lymphoma is usually rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP) or dose-adjusted etoposide, prednisone, vincristine, cyclophos It consists of pamide, doxorubicin and rituximab (DA-EPOCH-R).

Most lymphomas initially respond to either of these therapies, but tumors usually recur and eventually become refractory. As the number of regimens a patient receives increases, the resistance of the disease to chemotherapy increases. The average response to first line treatment is approximately 75%, second line treatment 60%, third line treatment 50%, and fourth line treatment about 35-40%. A response rate reaching 20% with a single agent in multiple relapsed sites is considered a definitive and warranted future study.

Current chemotherapeutic agents induce their antitumor response by inducing apoptosis through various mechanisms. However, many tumors eventually become resistant to these drugs. Bcl-2 and Bcl-X<i>_L</i>has been shown to confer chemotherapy resistance in vitro and more recently in short-term survival assays in vivo. These are Bcl-2 and Bcl-X<i>_L</i>If improved therapies can be developed aimed at inhibiting the function of

Bcl-2 and Bcl-X<i>_L</i>Apoptosis-promoting drugs targeting Bcl-2 family proteins, such as, are best administered according to a regimen that provides continuous, eg daily, supplementation of plasma concentrations to maintain concentrations in the therapeutically effective range. This can be achieved by parenteral, eg, intravenous (iv) or intraperitoneal (ip) administration on a daily basis. However, parenteral administration on a daily basis is often not practical in the clinical setting, especially for outpatients. To enhance the clinical utility of apoptosis-promoting agents as chemotherapeutic agents, for example in cancer patients, dosage forms with acceptable oral bioavailability or dosage forms that are less restrictive than solution formulations would be highly desirable. This dosage form and its regimen for oral administration is a significant advance in the treatment of many cancer types, including non-Hodgkin's lymphoma, and may more readily realize combination therapy with chemotherapeutic agents.

Summary of the invention

D less than 3₉₀There is provided a liquid pharmaceutical composition comprising an aqueous medium in which a solid particle compound having a particle size is suspended, wherein the compound is a compound of formula (I), a pharmaceutically acceptable salt thereof, a prodrug, a salt of a prodrug, or a salt thereof metabolite, wherein the aqueous medium comprises one or more pharmaceutically acceptable surfactants and one or more pharmaceutically acceptable basifying agents in amounts effective to together inhibit particle size increase.

Formula I

<img file="KR20120052937A_D0003.tif" />

In the above formula (I),

X³is chloro or fluoro,

(1) X⁴is azepan-1-yl, morpholin-4-yl, 1,4-oxazepan-4-yl, pyrrolidin-1-yl, -N (CH₃)₂, -N(CH₃)(CH(CH₃)₂), 7-azabicyclo[2.2.1]heptan-7-yl or 2-oxa-5-azabicyclo[2.2.1]hept-5-yl, R⁰silver <img file="KR20120052937A_D0004.tif" />

(where X⁵is -CH₂_-, -C(CH₃)_2- or -CH₂CH₂_-ego,

X⁶ and X⁷are both -H or both are methyl,

X⁸is fluoro, chloro, bromo or iodo);

(2) X⁴is azepan-1-yl, morpholin-4-yl, pyrrolidin-1-yl, -N (CH₃)(CH(CH₃)₂) or 7-azabicyclo[2.2.1]heptan-7-yl, R⁰silver <img file="KR20120052937A_D0005.tif" />(where X⁶, X⁷ and X⁸is as described above);

(3) X⁴is morpholin-4-yl or -N (CH₃)₂and R⁰silver <img file="KR20120052937A_D0006.tif" />(where X⁸is as described above).

Although the compositions of the present invention are primarily intended for oral administration, they are generally also suitable for other routes, including parenteral routes.

Furthermore, D of about 3 μm or less₉₀a compound of formula (I) or a pharmaceutically acceptable salt, a prodrug, a salt of a prodrug, or a metabolite thereof in a particulate form having a particle size; and a pharmaceutically acceptable excipient comprising (a) at least one surfactant and at least one basifying agent and (b) at least one dispersing agent and a bulking agent, wherein the composition is dispersed in an aqueous medium A suspension may be obtained wherein the surfactant and the basifying agent are together present in an effective amount to inhibit the increase in particle size.

Still further, there is provided an active pharmaceutical ingredient (API) comprising a compound of formula I or a pharmaceutically acceptable salt, prodrug, salt of a prodrug or metabolite thereof, wherein at least one pharmaceutically used basifying agent is provided. D of about 3 μm or less of the API in the presence₉₀There is provided a method for preparing a pharmaceutical composition comprising wet milling to particle size to obtain a milled drug and suspending the milled drug substrate in an aqueous medium having one or more pharmaceutically acceptable surfactants, wherein the at least one basifying agent and at least one surfactant are present in the resulting suspension in an effective amount to together inhibit the increase in particle size.

According to any of the above embodiments, the drug compound or API is, for example, ABT-263 or a crystalline salt thereof, for example ABT-263. <i>bis</i>-Hydrogen chloride (ABT-263) <i>bis</i>-HCl) salt.

Also provided is a method for treating a disease characterized by apoptotic dysfunction and/or overexpression of an anti-apoptotic Bcl-2 family protein, the method comprising: apoptotic dysfunction and/or overexpression of an anti-apoptotic Bcl-2 family protein It comprises orally administering to a subject having a disease characterized by a therapeutically effective amount of a composition as described above, for example, a composition comprising ABT-263 free base or ABT-263 bis-HCl. Examples of such diseases include many neoplastic diseases, including cancer. A particular exemplary type of cancer that can be treated according to the methods of the present invention is non-Hodgkin's lymphoma (NHL). Another particular exemplary type of cancer that can be treated according to the methods of the present invention is chronic lymphocytic leukemia. Another particular exemplary type of cancer that can be treated according to the methods of the invention is, for example, acute lymphocytic leukemia in pediatric patients.

Further, a method of maintaining a therapeutically effective plasma concentration of ABT-263 and/or one or more metabolites thereof in the bloodstream of a human cancer patient, eg, a patient with non-Hodgkin's lymphoma, chronic lymphocytic leukemia or acute lymphocytic leukemia. Also provided, wherein the method comprises administering a composition comprising the ABT-263 free base or crystalline salt in an amount equivalent to about 50 to about 500 mg of ABT-263 free base per day, an average interval of dosing from about 3 hours to about 7 days and administering to the subject.

Additional aspects of the invention, including more specific aspects of those provided above, will be found in or will become apparent from the following detailed description.

1 is a comparative solution of ABT-263 bis-HCl in a composition of the present invention (formulation II) and a lipid medium (formulation C) for 24 hours following oral administration to a dog (non-fasting state, except as otherwise indicated) ABT-263 plasma concentrations are graphically depicted and described in Example 3.

The suspension composition according to the present application comprises a nano-sized solid particle drug compound. It has been found that drug nanoparticles in the suspensions described herein do not appreciably aggregate, but yield a product in a stable formulation.

Unless otherwise stated herein, the term "nanoparticle" refers to a particle size (the diameter of the longest dimension of a particle) of about 3 μm (3,000 nm) or less. As used herein, "nanoparticles" include submicrons as well as micron-sized particles from about 1 to about 3 μm. Similarly, the adjective "nano-sized" is defined as the above nanoparticles. Unless otherwise stated in the text, "nanoparticulate" is a suspension or other composition, and "nanosuspension" is a D of about 3 μm or less.₉₀means having a particle size.

D of the composition₉₀Particle size is a parameter such that the 90% volume percentage of particles in the composition is smaller in their longest dimension than that parameter measured by conventional particle sizing techniques known to those skilled in the art. Such techniques include, for example, sedimentation field flow fractionation, photon correlation spectroscopy, light scattering and disk centrifugation. In various embodiments of the present invention, the suspensions are D₉₀The particle size is about 3,000 nm, about 2,000 nm, about 1,500 nm, about 1,000 nm, about 900 nm, about 800 nm, about 700 nm, about 600 nm, about 500 nm or less.

D of the composition₅₀ Particle size is a parameter such that the 50% volume percentage of the particles in the composition is smaller in their longest dimension than that parameter measured by conventional particle sizing techniques known to those skilled in the art. therefore D₅₀ Particle size is a measure of the particle size of the median volume, but sometimes refers to the "average or mean" particle size. In various embodiments of the invention, the suspensions are D₅₀The particle size is about 1,000 nm, about 900 nm, about 800 nm, about 700 nm, about 600 nm, about 500 nm, about 400 nm, about 350 nm, about 300 nm or less.

In a particular embodiment the suspension of the invention comprises D₉₀The particle size is about 1,000 nm or less and D₅₀ The particle size is about 400 nm or less. In another specific embodiment, the suspension of the invention comprises D₉₀The particle size is about 800 nm or less and D₅₀ The particle size is about 350 nm or less.

"Low solubility" and "poorly soluble" are those having a solubility in water of about 100 μg/ml or less. The present invention may be particularly advantageous as a drug that is essentially insoluble in water, ie has a solubility lower than about 10 μg/ml. Without wishing to be bound by theory, the advantages of nanoparticulate suspensions for such drugs lead in part to improved solubility according to the Kelvin equation, as well as improved dissolution rate proportional to surface area according to the well-known Whitney-Neuy equation. It could potentially reduce food effect as well as improve bioavailability.

It is recognized that the aqueous solubility of many compounds is PH-dependent, and it will be appreciated that the solubility of such compounds of interest herein is at a physiologically relevant pH, for example a pH of about 1 to about 8. Thus, in various embodiments, the drug has an aqueous solubility of less than about 100 μg/ml, such as less than about 30 μg/ml or less than about 10 μg/ml, at at least one point in the pH range of about 1 to about 8. have Illustratively, ABT-263 has a solubility in water of less than 4 μg/ml at pH 2.

In the composition of the present invention, the drug compound is a compound of formula (I) specified above, or a pharmaceutically acceptable salt, prodrug, salt of a prodrug, or a metabolite thereof.

In a further embodiment, the compound is X³It has the formula (I) which is fluoro.

In yet a further embodiment, said compound is X⁴is morpholin-4-yl.

In yet a further embodiment, said compound is R⁰this <img file="KR20120052937A_D0007.tif" />has the formula (I) wherein X⁵is -O-, -CH₂_-, -C(CH₃)₂- or -CH₂CH₂-and X⁶ and X⁷is both -H or both are methyl, X⁸is fluoro, chloro, bromo or iodo). Illustratively, according to this aspect, X⁵is -C(CH₃)₂-can be and/or X⁶ and X⁷may each be -H and/or X⁸may be chloro.

In yet a further embodiment, said compound is R⁰this <img file="KR20120052937A_D0008.tif" />has the formula (I) wherein X⁵is -O-, -CH₂_-, -C(CH₃)₂- or -CH₂CH₂-and X⁶ and X⁷is both -H or both are methyl, X⁸is fluoro, chloro, bromo or iodo). Illustratively, according to this aspect, X⁵is -C(CH₃)₂-can be and/or X⁶ and X⁷may each be -H and/or X⁸may be chloro.

In yet a further embodiment, said compound is X³is fluoro and X⁴is morpholin-4-yl.

In yet a further embodiment, said compound is X³is fluoro and R⁰this <img file="KR20120052937A_D0009.tif" />has the formula (I) wherein X⁵is -O-, -CH₂-, -C(CH₃)₂ -or -CH₂CH₂- and X⁶ and X⁷is both -H or both are methyl, X⁸is fluoro, chloro, bromo or iodo). Illustratively, according to this aspect, X⁵is -C(CH₃)₂-can be and/or X⁶ and X⁷may each be -H and/or X⁸may be chloro.

In yet a further embodiment, said compound is X⁴is morpholin-4-yl and R⁰this <img file="KR20120052937A_D0010.tif" />has the formula (I) wherein X⁵is -O-, -CH₂-, -C(CH₃)₂- or -CH₂CH₂- and X⁶ and X⁷is both -H or both are methyl, X⁸is fluoro, chloro, bromo or iodo). Illustratively, according to this aspect, X⁵is -C(CH₃)₂-can be and/or X⁶ and X⁷may each be -H and/or X⁸may be chloro.

In yet a further embodiment, said compound is X³is fluoro and X⁴is morpholin-4-yl and R⁰this <img file="KR20120052937A_D0011.tif" />has the formula (I) wherein X⁵is -O-, -CH₂-, -C(CH₃)₂ -or -CH₂CH₂- and X⁶ and X⁷is both -H or both are methyl, X⁸is fluoro, chloro, bromo or iodo). Illustratively, according to this aspect, X⁵is -C(CH₃)₂- or/or X⁶ and X⁷may each be -H and/or X⁸may be chloro.

The compounds of formula (I) may contain asymmetrically substituted carbon atoms in the R- or S-configuration, and these compounds may exist as a racemate, or in one configuration in excess of the other, for example It may be present in an enantiomeric ratio of at least about 85:15. The compound may be substantially enantiomerically pure, for example, may have an enantiomer ratio of at least about 95:5 or in some cases at least about 98:2 or at least about 99:1.

The compounds of formula (I) may alternatively or additionally contain a carbon-carbon double bond or a carbon-nitrogen double bond in the Z- or E-configuration, wherein the term "Z" means that large substituents are on the same side of the double bond. refers to the arrangement on the opposite side of the double bond, and the term "E" refers to the arrangement in which the large substituents are on opposite sides of the double bond. The compound may alternatively exist as a mixture of the Z-isomer and the E-isomer.

The compounds of formula (I) may alternatively or additionally exist as tautomers or equilibrium mixtures thereof in which a proton is transferred from one atom to another. Examples of tautomers include, by way of illustration, keto-enol, phenol-keto, oxime-nitroso, nitro-acy, imine-enamine and the like.

In some embodiments, the compound of formula (I) is present in nanoparticulate suspension, either alone in its parent compound form or together with the compound in salt or prodrug form.

The compounds of formula (I) may form acid addition salts, base addition salts or zwitterions. Salts of compounds of formula (I) may be prepared while the compounds are isolated or after purification. Acid addition salts are those derived from the reaction of a compound of formula (I) with an acid. For example, of the compounds of formula (I), acetate, adipate, alginate, bicarbonate, citrate, aspartate, benzoate, benzenesulfonate (besylate), bisulfate, butyrate, camphorate, cam Phorsulfonate, digluconate, formate, fumarate, glycerophosphate, glutamate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, lactobionate, lactate, maleate Acid salt, mesitylene sulfonate, methane sulfonate, naphthylene sulfonate, nicotinate, oxalate, pamoate, pectate, persulfate, phosphate, picrate, propionate, succinate, tartrate, thiocyanate, triclo Salts including roacetate, trifluoroacetate, para-toluenesulfonate and undecanoate may be used in the compositions of the present invention. Base addition salts, including those derived from the reaction of compounds with bicarbonates, carbonates, hydroxides or phosphates of cations such as lithium, sodium, potassium, calcium and magnesium, may likewise be used.

The compound of formula (I) usually has more than one protic nitrogen atom and thus more than 1 equivalent, for example from about 1.2 to about 2 equivalents, from about 1.5 to about 2 equivalents, or from about 1.8 to about 1 equivalent of the compound in question. Acid addition salts can be formed with 2 equivalents of acid.

ABT-263 can likewise form acid addition salts, base addition salts or zwitterions. A salt of ABT-263 can be prepared while the compound is isolated or after purification. Acid addition salts derived from the reaction of ABT-263 with acids include those listed above. Base addition salts, including those listed above, may likewise be used. ABT-263 has two or more protic nitrogen atoms, and thus more than 1 equivalent, for example from about 1.2 to about 2 equivalents, from about 1.5 to about 2 equivalents, or from about 1.8 to about 2 equivalents of an acid for 1 equivalent of the compound together can form acid addition salts.

Illustratively, for ABT-263, for example, bis-hydrochloride (<i>bis</i>-HCl) and bis-hydrobromide (<i>bis</i>-HBr) salts, including bis-salts, may be formed.

For example, ABT-263 having a molecular weight of 1047.5 g/mol and represented by the following structural formula <i>bis</i>-HCl can be prepared by a variety of methods, for example, a method that can be summarized as follows:

<img file="KR20120052937A_D0012.tif" />

ABT-263 free base may illustratively be prepared as described in Example 1 of the aforementioned 'US Patent Application Publication No. 2007/0027135, which is incorporated herein by reference in its entirety. A suitable weight of ABT-263 free base is dissolved in ethyl acetate. A solution of hydrochloric acid in ethanol (eg, about 4.3 kg of HCl in 80 g of ethanol) was mixed with at least 2 moles of HCl per mole of ABT-263 and the resulting ABT-263<i>bis</i>-Add to the ABT-263 solution in an amount to provide sufficient ethanol (at least about 20 vol) for crystallization of the -HCl salt. The solution is heated to about 45[deg.] C. with stirring and the seeds are added as a slurry in ethanol. After about 6 hours, the resulting slurry is cooled to about 20[deg.] C. over about 1 hour and mixed at this temperature for about 36 hours. Filter the slurry to ABT-263<i>bis</i>A crystalline solid, which is an ethanol solvate of -HCl, is recovered. The solid was dried with gentle stirring in vacuum and nitrogen for about 8 days to give a white, desolvated ABT-263 color.<i>bis</i>-HCl crystals are obtained. This material is suitable as an API for preparing the composition of the present invention.

Although it is recognized that the parent compound is strictly zwitterionic and thus does not always behave as a true base, the term "free base" is used herein for convenience to refer to the parent compound.

Compounds of formula (I) and methods for preparing such compounds are disclosed in US Patent Application Publication No. 2007/0027135 and/or the referenced US Patent Application Publication No. 2007/0072860, each of which is incorporated herein by reference in its entirety. . The terms for substituents as used herein are defined exactly as in the above publications.

In compounds of formula I with -NH, -C(O)OH, -OH or -SH residues, free -NH, -C(O)OH, - Prodrug-forming moieties can be attached which can release the parent compound with OH or -SH moieties. Salts of prodrugs may also be used.

Without wishing to be bound by theory, the therapeutic efficacy of the compounds of formula (I) is at least in part because they occupy the BH3 binding groove of the Bcl-2 family proteins such as Bcl-2, BCI-XL or Bcl-w, whereby the - It is thought to be due to the ability to bind to the protein in a manner that inhibits apoptotic action. In general, it will be found desirable to select compounds having a high binding affinity for the Bcl-2 family protein, for example, a Ki of about 5 nM or less, preferably 1 nM or less.

Nanoparticulate suspensions comprise a compound of formula (I) or a salt, a prodrug, a salt of a prodrug or a metabolite thereof as an individual solid state phase, which may be crystalline, semi-crystalline or amorphous. In the case of ABT-263, the free base form prepared according to the '135 publication is amorphous or glassy, which is usually a crystalline salt form of the drug, e.g., ABT-263 bis-HCl is used to prepare a nanosuspension. It is preferable to do However, upon suspending the salt in the presence of a basifying agent such as sodium bicarbonate, the salt may be partially converted to the free base, and the resulting product in a solid phase becomes at least partially amorphous. Thus, in one embodiment the nanosuspension comprises ABT-263 free base, ABT-263 bis-HCl, or a mixture thereof. It is observed that drug particles in ABT-263 nanosuspensions have a remarkably high degree of physical stability in such nanosuspensions, despite the possibility that they are at least partially amorphous, which is illustrated in Example 2 below.

The present inventors have found that the nanoparticulate suspensions described herein provide the advantages of acceptable shelf life with physical stability, as well as the robustness of the manufacturing process preferred for commercial products.

The compound of formula (I) or salt, prodrug, salt of prodrug or metabolite thereof is present in the nanoparticulate suspension of the present invention in a therapeutically effective amount when the composition is administered to a subject in need thereof according to an appropriate regimen. Dosages administered herein are expressed as parent compound equivalents (free base equivalents) unless the context requires otherwise. Typically, a unit dose (amount administered at a time) that can be administered at an appropriate frequency, eg, twice a day to once a week, is from about 10 to about 1,000 mg, depending on the compound in question. When the frequency of administration is once a day (qd), the unit dose and the daily dose are the same. Illustratively, for example, when the drug is ABT-263, the unit dose is typically from about 25 to about 1,000 mg, more typically from about 50 to about 500 mg, such as about 50, about 100, about 150, about 200, about 250, about 300, about 350, about 400, about 450 or about 500 mg, the free base equivalent. Tablets wherein the dosage form comprises a capsule shell or solid form enclosing the nanoparticulate composition in suspension, or a tablet comprising the nanoparticulate composition in a solid form, wherein the unit dose is a single capsule or tablet, or a plurality of capsules or tablets, most typically It may be delivered as 1 to about 10 capsules or tablets.

The higher the unit dose, the more desirable it is to select a suspension with a relatively high concentration of drug in solution. Typically, the drug concentration of the suspension is at least about 10 mg/ml, for example, from about 10 to about 500 mg/ml, although in certain instances lower or higher concentrations are acceptable or achievable. Illustratively, for example, where the drug is ABT-263, in various embodiments the drug concentration is at least about 10 mg/ml based on free base equivalent weight, e.g., from about 10 to about 400 mg/ml, or about 20 mg/ml or more, such as about 20 to about 200 mg/ml, such as about 20, about 25, about 30, about 40, about 50, about 75, about 100, about 125, about 150 or about 200 mg/ml.

The compositions of the present invention have good storage stability. In particular, they are physically stable so at least they do not have an unacceptable tendency to increase in particle size over time, for example through particle agglomeration. Particle agglomeration is a common problem in nanoparticulate suspensions. Surface modifiers, such as surfactants, are important in reducing the tendency of nanoparticles to agglomerate, and without being bound by theory, it is believed that one or more surfactants, if present in the composition of the present invention, help it.

Any "basifying agent" herein is any agent that raises the pH of the suspension medium. Any pharmaceutically acceptable basifying agent may be used, including without limitation hydroxides and bicarbonates of alkali metals such as sodium and potassium. In particular, sodium bicarbonate is shown by reference herein, but in some cases sodium bicarbonate may be substituted with other basifying agents.

The amount of sodium bicarbonate useful in the compositions of the present invention has no slight threshold and one of ordinary skill in the art can readily optimize the amount of a particular composition through routine storage-stability testing, for example. In general, good results can be obtained with amounts of sodium bicarbonate from about 20 to about 200 mg/ml, for example from about 40 to about 160 mg/ml.

The choice and amount of surfactant is not in the slightest critical and depends on the particular drug compound to be formulated and the desired degree of drug loading. Non-limiting examples of surfactants, individually or in combination, include quaternary (quaternary) ammonium compounds such as benzalkonium chloride, benzenetonium chloride and cetylpyridinium chloride; dioctyl sodium sulfosuccinate; polyoxyethylene alkylphenyl ethers such as nonoxynol 9, nonoxynol 10 and octoxynol 9; poloxamers (polyoxyethylene and polyoxypropylene block copolymers) such as poloxamer 188 and poloxamer 237; polyoxyethylene fatty acid glycerides and oils such as polyoxyethylene (8) caprylic/capric mono- and diglycerides, polyoxyethylene (35) castor oil and polyoxyethylene (40) hydrogenated castor oil; polyoxyethylene alkyl ethers such as cethes-10, laureth-4, laureth-23, oleth-2, oleth-10, oleth-20, steareth-2, steareth-10, steareth Areth-20, Steareth-100 and polyoxyethylene (20) cetostearyl ether; polyoxyethylene fatty acid esters such as polyoxyethylene (20) stearate, polyoxyethylene (40) stearate and polyoxyethylene (100) stearate; sorbitan esters such as sorbitan monolaurate, sorbitan monooleate, sorbitan monopalmitate and sorbitan monostearate; polyoxyethylene sorbitan esters such as polysorbate 20 and polysorbate 80; propylene glycol fatty acid esters such as propylene glycol laurate; sodium lauryl sulfate; fatty acids and salts thereof, such as oleic acid, sodium oleate and triethanolamine oleate; glyceryl fatty acid esters such as glyceryl monooleate, glyceryl monostearate and glyceryl palmitostearate; α-tocopheryl polyethylene glycol succinic acid (TPGS); thyroxapol and the like. In one embodiment, the at least one surfactant is a poloxamer or a mixture of poloxamers. Poloxamer 188 is a specific example. One or more surfactants typically make up a total of about 10 to about 100 mg/ml. In the case of poloxamer 188, an illustratively suitable amount is from about 10 to about 100 mg/ml, for example from about 15 to about 60 mg/ml.

The aqueous medium of the suspension may take the form of an injectable fluid such as water, saline (eg phosphate-buffered saline or PBS), or an immobilizing liquid such as fruit juice or soda. In one embodiment of the nanoparticulate drug compound, at least one surfactant and at least one basifying agent (and optionally additional components) are for reconstitution with a suitable aqueous medium to form a suspension composition of the invention immediately prior to use. It is prepared as a dry powder mixture. Such reconstituted powders may additionally contain, in addition to the above-mentioned ingredients, at least one pharmaceutically acceptable dispersing or bulking agent, usually a sugar, such as dextrose, mannitol or dextran. water-soluble substances, such as phosphate salts such as sodium or potassium phosphate; It must contain an organic acid, for example citric acid or tartaric acid, or a salt thereof or a mixture of these substances. Alternatively, the dry powder mixture may be administered to a subject for resuspension of nanoparticles in gastrointestinal fluid, for which administration such powder mixture may optionally be tableted or filled into capsules.

In the case of compounds of formula (I), it may be desirable to provide formulations that are chemically as well as physically stable. In particular, such formulations should not exhibit unacceptable oxidative degradation of the compounds of formula (I), for example at the thioether linkage of the (phenylsulfanyl)methyl group.

In this regard, compositions of the present invention comprising a compound of formula I, such as ABT-263 free base, ABT-263 bis-HCl or combinations thereof, are described in the previous '135 publication or reference [Tse et al. (2008), supra, has significant advantages over the solution compositions of ABT-263. It is believed that the solid phase form (crystalline, semi-crystalline or amorphous) of ABT-263 present in the nanosuspensions presented herein is significantly more resistant to oxidative degradation than ABT-263 in solution.

However, in some cases the tendency to oxidative degradation can be further reduced by including suitable antioxidants in the suspension composition.

A compound having "antioxidant" or "antioxidant" properties is a chemical compound capable of inhibiting, preventing, reducing or delaying the oxidation of another chemical or itself. Antioxidants can improve the stability and shelf life of lipid formulations as described herein, for example by inhibiting, preventing, reducing or delaying oxidation of a compound of Formula I in the formulation.

Improvements in stability or shelf life can be assessed, for example, by observing the rate of appearance or increase in the sulfoxide in the formulation. The sulfoxide in its entirety can be observed through repeated sampling and analysis, alternatively the samples are prepared for compounds of formula I, i.e.

<img file="KR20120052937A_D0013.tif" />A compound having (wherein X³, X⁴ and R⁰is as described above) sulfoxide decomposition products; or the formula

<img file="KR20120052937A_D0014.tif" />It can be analyzed more specifically for the sulfoxide degradation product of ABT-263 with

It will be understood that the sulfoxide degradation products referred to herein include both diastereomers at the stereocenter of the sulfur atom of the sulfoxide group.

As used herein, an "antioxidant effective amount" of an antioxidant is used in a formulation containing said antioxidant as compared to an otherwise similar formulation that does not contain the antioxidant,

(a) a significant reduction in the formation or accumulation of degradation products, e.g., the sulfoxide degradation products (e.g., at least about 25%, at least about 50%, at least about 75%, at least about 80%, at least about 85%) or a reduction of at least about 90%) and/or

(b) a significant increase in the time it takes for decomposition products to reach a threshold level (eg, at least about 30, at least about 60, at least about 90, or at least about 180 days)

is the amount that provides Storage stability studies to determine the extent of (a) a decrease in the formation or accumulation of degradation products or (b) an increase in the time it takes for the degradation products to reach a critical point in the formulation can be conducted at any suitable temperature or range of temperatures. have. Illustratively, a study at 5° C. may indicate storage stability under refrigerated conditions, a study at about 20-25° C. may indicate storage stability under normal ambient conditions, and a study at a temperature of about 30° C. or higher. may be useful in accelerated aging studies. Any suitable threshold of decomposition products may be selected as the endpoint, for example within the range of about 0.2% to about 2% of the initial amount of the compound of formula (I) present.

In various exemplary embodiments, when the antioxidant is stored under ambient conditions (eg about 20-25° C.) in a sealed container that is impermeable to ultraviolet light, the oxidative degradation of the drug in the formulation (the stated storage period is after completion, e.g. as measured by the amount of sulfoxide decomposition products present),

(a) less than about 1% for at least about 3 months;

(b) less than about 1% for at least about 6 months;

(c) less than about 1% for at least about 1 year;

(d) less than about 0.5% for at least about 3 months;

(e) less than about 0.5% for at least about 6 months; or

(f) less than about 0.5% for at least about 1 year;

It is included in an amount effective to maintain.

Antioxidants used in pharmaceutical compositions are most commonly agents that inhibit the generation of oxidizing species such as triplet or singlet oxygen, superoxide, peroxide and free hydroxyl radicals, or remove these oxidizing species when they occur. It is a drug that Examples of these classes of antioxidants commonly used include butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), retinyl palmitate, tocopherol, propyl gallate, ascorbic acid and ascorbyl palmitate. is included However, without wishing to be bound by theory, it is believed that the antioxidants useful herein function primarily as competitive substrates, i.e. "sacrificial" antioxidants that are preferentially attacked by oxidizing species and thus protect the drug from excessive degradation. It is an antioxidant.

In some embodiments, HCA is one or more antioxidant compounds of Formula II, pharmaceutically acceptable salts thereof, or Y¹is S and R³When H is, -SS- dimer thereof or a pharmaceutically acceptable salt of said dimer is included.

Formula II

<img file="KR20120052937A_D0015.tif" />

In the above formula (II),

n is 0, 1 or 2,

Y¹is S or Se,

Y²is the NHR¹, OH or H, wherein R¹is alkyl or alkylcarbonyl,

Y³Silver COOR² or CH₂OH, where R²is H or alkyl,

R³is H or alkyl;

wherein the alkyl groups are optionally substituted independently by one or more substituents independently selected from the group consisting of carboxyl, alkylcarbonyl, alkoxycarbonyl, amino and alkylcarbonylamino.

In another embodiment, HCA is an antioxidant compound of Formula III.

Formula III

<img file="KR20120052937A_D0016.tif" />

In the above formula (III),

Y is S, Se or SS;

R⁴ and R⁵is H, alkyl and (CH₂)_nR⁶independently selected from, wherein n is 0 to 10, and R⁶is arylcarbonyl, alkylcarbonyl, alkoxycarbonyl, carboxyl or CHR⁷R⁸-substituted alkyl, wherein R⁷ and R⁸is independently CO₂R⁹, CH₂OH, hydrogen or NHR¹⁰, where R⁹is H, alkyl, substituted alkyl or arylalkyl, R¹⁰is hydrogen, alkyl, alkylcarbonyl or alkoxycarbonyl.

An "alkyl" or "alkoxy" group that forms part of an "alkyl" substituent or a substituent according to Formula II or Formula III has from 1 to about 18 carbon atoms and may be straight-chain or branched.

An "aryl" group forming part of a substituent according to formula (III) is a phenyl group, unsubstituted or substituted with one or more hydroxy, alkoxy or alkyl groups.

In some embodiments, R in Formula II¹silver C₁_-4 Alkyl (such as methyl or ethyl) or (C_1-4 alkyl)carbonyl (eg acetyl).

In some embodiments, R in Formula II²is H or C₁_-18 Alkyl such as methyl, ethyl, propyl (such as n-propyl or isopropyl), butyl (such as n-butyl, isobutyl or t-butyl), octyl (such as n-octyl or 2-ethylhexyl) , dodecyl (eg lauryl), tridecyl, tetradecyl, hexadecyl or octadecyl (eg stearyl).

R³is usually H or C₁_-4 alkyl (eg methyl or ethyl).

The HCA may be, for example, a natural or synthetic amino acid or a derivative such as an alkyl ester thereof or an N-acyl derivative, or a salt of the amino acid or a derivative thereof. When the amino acid or derivative thereof is derived from a natural source, it is usually in the L-configuration, but it is understood that the D-isomer and mixture of D,L-isomers may be substituted if desired.

Non-limiting examples of HCA useful herein include β-alkylmercaptoketone, cysteine, cystine, homocysteine, methionine, thiodiglycolic acid, thiodipropionic acid, thioglycerol, selenocysteine, selenomethionine and salts, esters thereof, amides and thioethers, and combinations thereof. More specifically, the at least one HCA is N-acetylcysteine, N-acetylcysteine butyl ester, N-acetylcysteine dodecyl ester, N-acetyl-cysteine ethyl ester, N-acetylcysteine methyl ester, N-acetylcysteine octyl ester, N-Acetyl-cysteine propyl ester, N-acetylcysteine stearyl ester, N-acetylcysteine tetradecyl ester, N-acetylcysteine tridecyl ester, N-acetylmethionine, N-acetylmethionine butyl ester, N-acetylmethionine dodecyl Ester, N-acetylmethionine ethyl ester, N-acetylmethionine methyl ester, N-acetylmethionine octyl ester, N-acetylmethionine propyl ester, N-acetylmethionine stearyl ester, N-acetylmethionine tetradecyl ester, N-acetylmethionine tridecyl ester, N-acetylselenocysteine, N-acetylselenocysteine butyl ester, N-acetylselenocysteine dodecyl ester, N-acetylselenocysteine ethyl ester, N-acetylselenocysteine methyl ester, N-acetylselenocysteine octyl ester, N-acetylselenocysteine propyl ester, N-acetylselenocysteine stearyl ester, N-acetylselenocysteine tetradecyl ester, N-acetyl Selenocysteine tridecyl ester, N-acetylselenomethionine, N-acetylselenomethionine butyl ester, N-acetylselenomethionine dodecyl ester, N-acetylselenomethionine ethyl ester, N-acetylselenomethionine methyl ester , N-acetylselenomethionine octyl ester, N-acetylselenomethionine propyl ester, N-acetylselenomethionine stearyl ester, N-acetylselenomethionine tetradecyl ester, N-acetylselenomethionine tridecyl ester, cysteine , cysteine butyl ester, cysteine dodecyl ester, cysteine ethyl ester, cysteine methyl ester, cysteine octyl ester, cysteine propyl ester, cysteine stearyl ester, cysteine tetradecyl ester, Cysteine tridecyl ester, cystine, cystine dibutyl ester, cystine di(dodecyl) ester, cystine diethyl ester, cystine dimethyl ester, cystine dioctyl ester, cystine dipropyl ester, cystine distearyl ester, cystine di(tetradecyl) ) ester, cystine di(tridecyl) ester, N,N-diacetylcystine, N,N-diacetylcystine dibutyl ester, N,N-diacetylcystine diethyl ester, N,N-diacetylcystine di( dodecyl) ester, N,N-diacetylcystine dimethyl ester, N,N-diacetylcystine dioctyl ester, N,N-diacetylcystine dipropyl ester, N,N-diacetylcystine distearyl ester, N ,N-diacetylcystine di(tetradecyl) ester, N,N-diacetylcystine di(tridecyl) ester, dibutyl thiodiglycolate, dibutyl thiodipropionate, di(dodecyl)thiodiglycol Late, di(dodecyl) thiodipropionate, diethyl thiodiglycolate, diethyl thiodipropionate, Dimethyl thiodiglycolate, dimethyl thiodipropionate, dioctyl thiodiglycolate, dioctyl thiodipropionate, dipropyl thiodiglycolate, dipropyl thiodipropionate, distearyl thiodiglycolate , distearyl thiodipropionate, di(tetradecyl) thiodiglycolate, di(tetradecyl) thiodipropionate, homocysteine, homocysteine butyl ester, homocysteine dodecyl ester, homocysteine ethyl ester, homocysteine methyl ester, Homocysteine octyl ester, homocysteine propyl ester, homocysteine stearyl ester, homocysteine tetradecyl ester, homocysteine tridecyl ester, methionine, methionine butyl ester, methionine dodecyl ester, methionine ethyl ester, methionine methyl ester, methionine octyl ester, methionine propyl ester, methionine Methionine stearyl ester, methionine tetradecyl ester, methionine tridecyl ester, S-methylcysteine, S-Methyl-cysteine butyl ester, S-methylcysteine dodecyl ester, S-methylcysteine ethyl ester, S-methyl-cysteine methyl ester, S-methylcysteine octyl ester, S-methylcysteine propyl ester, S-methyl-cysteine Stearyl ester, S-methylcysteine tetradecyl ester, S-methylcysteine tridecyl ester, selenocysteine, selenocysteine butyl ester, selenocysteine dodecyl ester, selenocysteine ethyl ester, selenocysteine methyl ester, selenocysteine Nocysteine octyl ester, selenocysteine propyl ester, selenocysteine stearyl ester, selenocysteine tetradecyl ester, selenocysteine tridecyl ester, selenomethionine, selenomethionine butyl ester, selenomethionine dodecyl ester, selene nomethionine ethyl ester, selenomethionine methyl ester, selenomethionine octyl ester, selenomethionine propyl ester, selenomethionine stearyl ester, selenomethionine tetradecyl ester, selenomethionine tridecyl ester, thiodiglycolic acid, thiodipropionic acid, thioglycerol, isomers and mixtures of isomers thereof, and salts thereof.

Salts of HCA compounds are acetate, adipate, alginate, bicarbonate, citrate, aspartate, benzoate, benzenesulfonate (besylate), bisulfate, butyrate, camphorate, camphorsulfonate, di Gluconate, formate, fumarate, glycerophosphate, glutamate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, lactobionate, lactate, maleate, mesitylene sulfone Acid salt, methanesulfonate, naphthylenesulfonate, nicotinate, oxalate, pamoate, pectate, persulfate, phosphate, picrate, propionate, succinate, tartrate, thiocyanate, trichloroacetate, trichloroacetate acid addition salts such as fluoroacetate, para-toluenesulfonate and undecanoate. In certain embodiments, the hydrochloride salt of one of the individually mentioned compounds is present in the composition in an antioxidant effective amount.

Without wishing to be bound by theory, it is generally believed that heavy chalcogenide antioxidants such as those exemplified above protect the active compound by being oxidized preferentially over the drug compound as they may themselves be more readily oxidized. In general, to provide an acceptable degree of protection to the drug compound in this mode of action, the antioxidant must be present in a substantial amount, for example in a molar ratio of at least about 1:10 to the drug compound. In some embodiments, the molar ratio of antioxidant to drug compound is from about 1:10 to about 2:1, such as from about 1:5 to about 1.5:1. Best results will sometimes be obtained when the molar ratio is approximately 1:1, i.e. from about 8:10 to about 10:8.

Another class of sulfur-containing antioxidants, sulfite inorganic antioxidants, bisulfites, metabisulfites and thiosulfates, may be useful in the compositions of the present invention. These antioxidants are used in aqueous solutions. Salts of sodium and potassium of the sulfites, bisulfites, metabisulfites and thiosulfates are useful antioxidants according to this embodiment; More particularly sodium and potassium metabisulfite. Such sulfur-containing antioxidants can be effective at concentrations that are much lower than providing a molar equivalent to the concentration of the drug compound, for example, when the molar ratio to drug compound is as low as 1 to 20 or less.

To minimize sulfoxide formation, a chelating agent such as EDTA or a salt thereof (eg, disodium EDTA or calcium disodium EDTA), optionally in an amount from about 0.002% to about 0.02% by weight of the composition is added Chelating agents sequester metal ions that can promote oxidative degradation.

Sulfoxide formation can be further minimized by selecting formulation components with low peroxide values. The peroxide value is a well-established property of pharmaceutical excipients and is generally expressed (as herein) in units (meq/kg) corresponding to milliequivalents of peroxide per kg of excipient. Some excipients have inherently low peroxide values, while others, for example those with unsaturated fatty acids such as oleyl moieties and/or polyoxyethylene chains, may be sources of peroxides.

Optional components of another suspension composition include buffering agents, coloring agents, flavoring agents, preservatives, sweetening agents, tonicifying agents, and combinations thereof.

In one embodiment of the present invention, an active pharmaceutical comprising a compound of formula (I) or a pharmaceutically acceptable salt, prodrug, salt of a prodrug or a metabolite thereof, for example ABT-263 or a crystalline salt thereof providing an ingredient (API), wherein said API in the presence of at least one basifying agent (such as sodium bicarbonate) has a D of about 3 μm or less₉₀A method is provided for preparing a pharmaceutical composition comprising wet milling to particle size to obtain a milled drug, and suspending the milled drug substrate in an aqueous medium having at least one surfactant, wherein at least one base The agent and at least one surfactant are present in the resulting suspension in an effective amount to together inhibit the increase in particle size.

Any suitable wet grinding method may be used. A wet milling method that has been found particularly useful is the high pressure homogenization state, for example, as described in Example 1 below.

The present invention is not limited to compositions prepared by any of the methods described herein. However, the composition prepared by the above method is a special aspect of the present invention.

In one embodiment, the method can further be reconstituted by adding at least one pharmaceutically acceptable dispersing or bulking agent to the suspension and drying the suspension (eg, freeze-dried or lyophilized or alternatively spray-dried). obtaining a dry powder and optionally making the powder into tablets (eg molded or compressed) or filling the powder into capsules to prepare unit dosage form.

In addition, the advantages of stabilizing sodium bicarbonate include wet processing to reduce particle size, e.g. D₉₀It has been found that particle sizes of up to about 700 nm are possible in the presence of sodium bicarbonate. Illustratively, when the same parameter treatment was performed without sodium bicarbonate as shown in Example 2 below, D₉₀ The particle size could not be reduced below 1,000 nm. The use of the wet grinding method in this method has the advantage of reducing the exposure of the API to high temperatures and reducing the risk of pyrolysis of the API compared to dry grinding. In one aspect, the processing temperature is controlled in the range of about 1 to about 5° C. of a target temperature of, for example, about 5° C. to about 30° C. This can be achieved by conventional means, such as operating the formulation by immersing the heat exchanger in a water bath with a cooler.

The composition may be prepared for wet milling at a final concentration, or it may be prepared by dilution to the desired concentration after high concentration and wet milling. At least one surfactant and optionally optional additional ingredients may be added before or after wet grinding.

The composition of the present invention is usually suitable for "oral delivery", ie, oral administration; However, such compositions may be useful for delivering drugs to a subject in need thereof by administration by other routes, including parenteral, sublingual, buccal, nasal, pulmonary, topical, transdermal, intradermal, ocular, otic, rectal, vaginal, intragastric, intracranial, intrasynovial and internal articular routes are included without limitation. In a particular embodiment the composition is used for oral and/or parenteral administration.

The terms "oral administration" and "administered orally" herein refer to administration by mouth (po) to a subject, ie, the composition is swallowed immediately, for example, with an appropriate volume of water or other beverage. "Oral administration" herein is distinguished from oral administration which does not include direct swallowing of the composition, for example, sublingual or buccal administration or topical administration to the oral epidermis such as periodontal tissue.

The unexpectedly discovered nanoparticulate ABT-263 bis HCl suspension of the present invention when administered orally exhibits enhanced enhancement compared to a standard solution of the drug, for example, a solution in a vehicle consisting of 10% DMSO in PEG-400 as disclosed in the '135 publication. Provides bioabsorption. In clinical trials (Example 3 below), it was found that the bioabsorption capacity corresponds to that obtained with a lipid solution formulation of ABT-263 bis-HCl (herein "Formulation C"). Such enhanced bioabsorption rate may be, for example, a pharmacokinetic (PK) profile or AUC (eg AUC) with one or more high Cmax.₀_-24or AUC₀_-∞) can be confirmed by the increased bioavailability measured by Illustratively, bioavailability can be calculated taking into account any differences between oral and intravenous (iv) doses, e.g., the AUC for oral delivery of a test compound as a percentage of the AUC for intravenous delivery of the drug in a suitable solution. Using the parameter F to calculate, it can be expressed as a percentage.

Bioavailability can be determined by studying PK in humans or any suitable model species. For this purpose, a dog model is generally suitable, as exemplarily described in Example 3 below. In various exemplary embodiments, when the drug is a crystalline salt of ABT-263, such as ABT-263 bis-HCl, the compositions of the present invention are administered in a single dose of about 2.5 to 10 mg/kg to fasting or non-fasting animals in a canine model. When administered in dose, oral bioavailability is at least about 15% or at least about 20% or at least about 25%, up to about 50% or greater.

Compositions encompassed herein, including those generally or specifically described herein, provide for oral delivery to a subject of a drug that is a compound of Formula I or a pharmaceutically acceptable salt, prodrug, prodrug of a salt, or a metabolite thereof useful for Accordingly, the methods of the present invention for such drug delivery to a subject include the oral administration compositions set forth above.

The subject may be a human or non-human (eg, a farm, zoo, work or companion animal or laboratory animal used as a model), but important aspects include, for example, apoptotic dysfunction and/or anti-apoptotic Bcl A human patient in need of a drug to treat a disease characterized by overexpression of the -2 family protein. Human subjects can be male or female and of any age, but typically adults.

The composition is usually administered in an amount to provide a therapeutically effective daily dose of the drug. As used herein, the term "daily dose" refers to an amount of a drug administered per day regardless of the frequency of administration. For example, if a subject receives a unit dose of 150 mg twice daily, the daily dose is 300 mg. It will be understood that the use of the term "daily dose" does not imply that a particular dosage must be administered once a day. However, in certain embodiments the frequency of administration is once daily (qb), and the unit dose and the daily dose are the same.

What constitutes a therapeutically effective amount depends on the particular compound, the subject (including the species and the subject's weight), the disease to be treated (eg, a particular type of cancer), the stage and/or severity of the disease, and the individual subject's tolerance to the compound. , depends on other factors, such as whether the compound is administered alone or in combination with one or more other drugs, eg, whether other chemotherapeutic agents are used to treat cancer. Therefore, the daily dose can vary within wide limits, for example from about 10 to about 1,000 mg. A large or small daily dose may be appropriate in certain circumstances. It is to be noted that a 'therapeutically effective' amount recited herein does not necessarily require that a drug herein be pharmaceutically effective only when administered in a single dose; typically, therapeutic efficacy will depend on the dosage regimen, including the appropriate frequency and duration of administration. depending on the repeatedly administered composition. While the selected daily dose should be sufficient to provide an advantage in terms of cancer treatment, it is highly desirable that it is not sufficient to cause unacceptable or intolerable side effects. A suitable therapeutically effective amount can be selected by a physician of ordinary skill without undue experimentation on the basis of those disclosed herein and recited herein, taking into account factors as noted above. For example, to reduce the risk of side effects, a physician may start a cancer patient treatment course with a relatively low daily dose, optimizing with increasing dose over a period of days or weeks or longer.

Illustratively, a suitable dose of ABT-263 is generally from about 25 to about 1,000 mg/day, more typically from about 50 to about 500 mg/day or from about 200 to about 400 mg/day, such as about 50, about 100, about 150, about 200, about 250, about 300, about 350, about 400, about 450 or about 500 mg/day, the average dosing interval being from about 3 hours to about 7 days, for example from about 8 hours to about 3 days or about 12 hours to about 2 days. For severe cases, once a day (qd) dosing regimen is suitable.

"Average dosing interval" is defined herein as a period of time divided by the number of unit doses administered over a period of time, for example a day or a week. For example, if a drug is administered 3 times a day, the average dosing period is 8 hours (24 hours divided by 3) around 8 am, noon and 6 pm. If the drug is tableted as separate dosage forms such as tablets or capsules, the number of dosage forms administered at one time (eg 2 to 10) should be considered for the purpose of the defined mean dosing interval as a unit dose.

For example, if the drug compound is ABT-263 in the form of ABT-263 bis-HCl, in some embodiments the daily dosage and dosing interval may be selected to maintain the ABT-263 plasma concentration between 0.5 and 10 μg/ml. can Therefore, during the course of treatment with ABT-263 in accordance with such embodiments, the steady-state peak plasma concentration (Cmax) should generally not exceed about 10 μg/ml, and the steady-state trough plasma concentration (Cmin) generally should not exceed about 0.5 μg/ml. It must not fall below ml. Within the ranges given above, it will be found desirable to select a daily dosage and average dosing interval effective to provide a Cmax/Cmin ratio of about 5 or less, eg, about 3 or less at steady state. It will be appreciated that the longer the dosing period, the higher the ratio Cmax/Cmin tends to be. Illustratively, an ABT-263 Cmax of about 3 to 8 μg/ml and a Cmin of about 1 to about 5 μg/ml at steady state may be targeted by the method. Steady-state values of Cmax and Cmin can be established in human PK studies performed according to standard protocols including, without limitation, those that may be approved by regulatory agencies such as the Food and Drug Administration (FDA).

Administration according to this embodiment can be done with or without food, ie, on an empty stomach or on an empty stomach. However, since the compositions of the present invention can exhibit positive phagocytosis, it is generally preferred to administer the compositions to patients on a non-fasting basis.

The compositions of the invention are suitable for single or combination therapy, for example in combination with other chemotherapy or ionizing radiation. A particular advantage of the present invention is that it permits oral administration once per day, and is a convenient regimen for patients under treatment with other orally administered drugs once per day. Oral administration can be easily accomplished by the patient himself/herself or by a caregiver at the patient's home; It is also a convenient route of administration for patients in hospitals or residential care facilities.

Combination therapy is illustratively a composition of the invention, for example a composition comprising ABT-263, bortezomib, carboplatin, cisplatin, cyclophosphamide, dacarbazine, dexamethasone, docetaxel, doxorubicin, etoposide, flu with one or more of darabine, irinotecan, paclitaxel, rapamycin, rituximab, vincristine and the like, for example CHOP (cyclophosphamide + doxorubicin + vincristine + prednisone), RCVP (rituximab + cyclophosphamide + vincristine + prednisone), R-CHOP (rituximab + CHOP) or DA-EPOCH-R (dose-adjusted etoposide, prednisone, vincristine, cyclophosphamide, doxorubicin and rituximab ), including administration with multi-therapeutic agents such as

For example, the composition of the present invention comprising ABT-263 may include, but is not limited to, an alkylating agent, an angiogenesis inhibitor, an antibody, an antimetabolites, a mitotic agent, an antiproliferative agent, an antiviral agent, aurora kinase inhibitors, other pro-apoptotic agents (eg Bcl-xL, Bcl-w and Bfl-1 inhibitors), activators of the death receptor pathway, Bcr-Abl kinase inhibitors, BiTE (bispecific T-cell engineer) antibodies, Antibody-drug conjugates, biological response modifiers, cyclin-dependent kinase (CDK) inhibitors, cell cycle inhibitors, cyclooxygenase-2 (COX-2) inhibitors, dual variable domain binding protein (DVD), human epidermal growth factor receptor 2 (EbB2 or HER/2neu) inhibitors, growth factor inhibitors, heat shock protein (HSP)-90 inhibitors, histone deacetylase (HDAC) inhibitors, hormone therapy agents, immune agents, apoptotic protein inhibitors (IAPs), intercalating antibiotics, kinase inhibitors, kinesin inhibitors, JAK2 inhibitors, Mammalian-targeted rapamycin (mTOR) inhibitors, microRNAs, mitogen-activated extracellular signal-regulated kinase (MEK) inhibitors, multivalent binding proteins, non-steroidal anti-inflammatory drugs (NSAIDs), poly-ADP (adenosine) diphosphate)-ribose polymerase (PARP) inhibitors, platinum chemotherapeutic agents, polo-like kinase (PLK) inhibitors, phosphoinositide-3 kinase (PI3K) inhibitors, proteasome inhibitors, purine homologs, pyrimidine analogues, Receptor thyroxine kinase inhibitors, retinoids, deltoids, plant alkaloids, small inhibitory ribonucleic acids (siRNAs), topoisomerase inhibitors, ubiquitin ligase inhibitors, and the like may be administered in combination therapy with one or more therapeutic agents.

BiTE antibodies are bispecific antibodies that direct T-cells to attack cancer cells by binding to both cells at the same time. T-cells attack target cancer cells. Examples of BiTE antibodies include, but are not limited to, adekatumumab (Micromet MT201), blinatumomab (Micromet MT103), and the like. Without wishing to be bound by theory, one of the mechanisms by which T-cells induce apoptosis of target cancer cells is by exocytosis of cytolytic granule components comprising perforin and granzyme B. In this regard, Bcl-2 appears to reduce the induction of apoptosis by both perforin and granzyme B. These data suggest that inhibition of Bcl-2 may increase the cytotoxic effect induced by T-cells when targeting cancer cells [Sutton et al. (1997) J. Immunol. 158:5783-5790]

siRNAs are molecules with endogenous RNA bases or chemically modified nucleotides. Such modifications do not abrogate cellular activity, but rather confer increased stability and/or increased cellular titer. Examples of chemical modifications include a phosphorothioate group, 2'-deoxynucleotide, 2'-OCH₃-containing ribonucleotides, 2'-F-ribonucleotides, 2'-methoxyethyl ribonucleotides, combinations thereof, and the like. siRNAs can have different lengths (e.g. 10 to 200 bps) and different structures (e.g. hairpins, single/double stranded, bulge, nick/gap, mismatch) and have active gene silencing. processed within the cell to provide Double-stranded siRNA (dsRNA) may have the same number of nucleotides on each strand (blunt ends) or may have asymmetric ends (overhangs). One or two nucleotide overhangs may be present on the sense and/or antisense strand as well as the 5- and/or 3-end of a given strand. For example, siRNA targeting Mcl-1 has been shown to enhance the activity of ABT-263 or ABT-737 in various cancer cell lines.<i>et</i><i></i><i>al</i><i>.</i> (2008) <u>Cancer</u><u></u><u>Res</u><u>.</u> 68:3421-3428 and references therein].

A multivalent binding protein is a binding protein comprising two or more antigen binding sites. Multivalent binding proteins are engineered to have three or more antigen binding sites and are generally not native antibodies. The term "multispecific binding protein" refers to a binding protein capable of binding two or more related or unrelated targets. Dual variable domain (DVD) binding proteins are tetravalent or multivalent binding proteins that bind to proteins comprising two or more antigen binding sites. Such DVDs may be monospecific (ie capable of binding one antigen) or multispecific (ie capable of binding more than one antigen). A DVD binding protein comprising two heavy chain DVD polypeptides and two light chain DVD polypeptides is called a DVD Ig. Each half of a DVD Ig contains a heavy chain DVD polypeptide, a light chain DVD polypeptide and two antigen binding sites. Each binding site comprises a heavy chain variable domain and a light chain variable domain, with a total of six CDRs involved in antigen binding per antigen binding site.

Alkylating agents include altretamine, AMD-473, AP-5280, apaziquione, bendamustine, brostalysin, busulfan, carboquione, carmustine (BCNU), chlorambucil, Cloretazine (Laromustine, VNP). 40101M), cyclophosphamide, dacarbazine, estramustine, fotemustine, gluphosphamide, ifosfamide, KW-2170, lomustine (CCNU), maposfamide, melphalan, mitobronitol, mito lactol, nimustine, nitrogen mustard N-oxide, ranimustine, temozolomide, thiotepa, threosulfan, troposphamide and the like.

As angiogenesis inhibitors, epidermal growth factor receptor (EGFR) inhibitors, endothelial-specific receptor thyroxine kinase (Tie-2) inhibitors, insulin growth factor-2 receptor (IGFR-2) inhibitors, matrix metalloproteinase-2 (MMP-) 2) inhibitors, matrix metalloproteinase 9 (MMP-9) inhibitors, platelet-derived growth factor receptor (PDGFR) inhibitors, thrombospondin homologues, vascular endothelial growth factor receptor tyrosine kinase (VEGFR) inhibitors, and the like.

Antimetabolites include Alimta (Permetrex Disodium, LY231514, MTA), 5-azacitidine, Xeloda (Capecitabine), Chamopher, Leustat (Cladribine), Clofarabine, Cytarabine , cytarabine oxphosphate, cytosine arabinoside, decitabine, deferoxamine, doxyfluridine, eflonithine, EICAR (5-ethynyl-1-β-D-ribofuranosilimidazole-4- carboxamide), enocitabine, ethenylcytidine, fludarabine, 5-fluorouracil (5-FU) alone or leucovorin, Gemzar (gemcitabine), hydroxyurea, Alkeran (melparan) ), mercaptopurine, 6-mercaptopurine riboside, methotrexate, mycophenolic acid, nerarabine, nolatrexd, oxphosphate, ferritresol, pentostatin, raltitrexd, ribavirin, S-1, triapine, trimetrexate, TS-1, thiazopurine, tegafu, vidarabine, UFT, and the like.

Antiviral agents include ritonavir, hydroxychloroquine, and the like.

Aurora kinase inhibitors include ABT-348, AZD-1152, MLN-8054, VX-680, Aurora A-specific kinase inhibitors, Aurora B-specific kinase inhibitors, Pan-Aurora kinase inhibitors, and the like.

Bcl-2 family protein inhibitors other than ABT-263 or the compound of formula (I) herein include AT-101 ((-)gosipol), Genasense Bcl-2-targeting antisense oligonucleotide (G3139 or Olimergen), IPI - 194, IPI-565, ABT-737, GX-070 (Ovatoclax), etc.

Bcr-Abl kinase inhibitors include dasatinib (BMS-354825), Gleevec (imatinib), and the like.

CDK inhibitors include AZD-5438, BMI-1040, BMS-387032, CVT-2584, flavopyridol, GPC-286199, MCS-5A, PD0332991, PHA-690509, celiciclib (CYC-202 or R-Roscoby). tin), ZK-304709, and the like.

COX-2 inhibitors are ABT-963, Arcoxia (Etoricoxib), Bextra (Valdecoxib), BMS-347070, Celebrex (Celecoxib), COX-189 (Lumiracoxib), CT-3 , Deramaxx (Deracoxib), JTE-522, 4-methyl-2- (3,4-dimethylphenyl) -l- (4-sulfamoylphenyl) -lH-pyrrole, MK-663 (ethoricoxib) ), NS-398, parecoxib, RS-57067, SC-58125, SD-8381, SVT-2016, S-2474, T-614, Vioxx (rofecoxib) and the like.

EGFR inhibitors include ABX-EGF, anti-EGFR immunoliposomes, EGF-vaccine, EMD-7200, Erbitux (cetuximab), HR3, IgA antibody, Iressa (gefitinib), Tarceva (erlotinib or OSI -774), TP-38, EGFR fusion protein, Tykerb (lapatinib), and the like.

ErbB2 receptor inhibitors include CP-724714, CI-1033 (canertinib), Herceptin (trastuzumab), Tykerb (lapatinib), Omnitarg (2C4, fetuzumab), TAK-165, GW-572016 (io napamib), GW-282974, EKB-569, PI-166, dHER2 (HER2 vaccine), APC-8024 (HER2 vaccine), anti-HER/2neu bispecific antibody, B7.her2IgG3, AS HER2 trifunctional dual specific antibodies, mAB AR-209, mAB 2B-1, and the like.

Histone deacetylase inhibitors include depsipeptide, LAQ-824, MS-275, trapocin, suberoylanilide hydroxamic acid (SAHA), TSA, valproic acid, and the like.

HSP-90 inhibitors include 17AAG, CNF-101, CNF-1010, CNF-2024, 17-DMAG, geldanamycin, IPI-504, KOS-953, Mycograb (human recombinant antibody to HSP-90), nab- 17AAG, NCS-683664, PU24FC1, PU-3, Radicicol, SNX-2112, STA-9090, VER-49009, and the like.

Apoptotic protein inhibitors include HGS-1029, GDC-0145, GDC-0152, LCL-161, LBW-242, and the like.

Antibody-drug conjugates are anti-CD22-MC-MMAF, anti-CD22-MC-MMAE, anti-CD22-MCC-DMl, CR-Ol 1-vcMMAE, PSMA-ADC, MEDI-547, SGN-19A, SGN- 35, SGN-75, and the like.

Death receptor pathway activators include TRAIL, and antibodies or other agents targeting TRAIL or death receptors (eg DR4 and DR5), eg, Apomab, Conatumumab, ETR2-ST01, GDC0145 (lexatumumab) ), HGS-1029, LBY-135, PRO-1762, trastuzumab, and the like.

Kinesin inhibitors include Eg5 inhibitors such as AZD-4877 and ARRY-520, CENPE inhibitors such as GSK-923295A, and the like.

JAK2 inhibitors include CEP-701 (resaurtinib), XL019, INCB-018424, and the like.

MEK inhibitors include ARRY-142886, ARRY-438162, PD-325901, PD-98059, and the like.

mTOR inhibitors include AP-23573, CCI-779, everolimus, RAD-001, rapamycin, temsirolimus, ATP-competitive TORC1/TORC2 inhibitors (including PI-103, PP242, PP30 and Thorin 1), etc. do.

Non-steroidal anti-inflammatory drugs (NSAIDs) include Amigesic (salsalate), Dolobid (diflunisal), Motrin (ibuprofen), Orudis (ketoprofen), Relafen (nabumetone), Feldene ( piroxicam), ibuprofen cream, Aleve and Naprosyn (naproxen), Voltaren (diclofenac), Indocin (indomethacin), Clinoril (Sulindac), Tolectin (tolmethine), Lodine (etodolac) ), Toradol (ketorolac), Daypro (oxaprozin), and the like.

PDGFR inhibitors include CP-673451, CP-868596, and the like.

Platinum chemotherapeutic agents include cisplatin, Eloxatin (oxaliplatin), eptaplatin, lovaplatin, nedaplatin, Paraplatin (carboplatin), picoplatin, satraplatin, and the like.

Polo-like kinase inhibitors include BI-2536 and the like.

Phosphoinositide-3 kinase (PI3K) inhibitors include wortmannin, LY-294002, XL-147, CAL-120, ONC-21, AEZS-127, ETP-45658, PX-866, GDC-0941, BGT226 , BEZ235, XL765, etc.

Thrombospondin homologs include ABT-510, ABT-567, ABT-898, TSP-I, and the like.

VEGFR inhibitors include Avastin (bevacizumab), ABT-869, AEE-788, Angiozyme (ribozymes that inhibit angiogenesis (Ribozyme Pharmaceuticals (Boulder, CO) and Syron (Emeryville, CA)), Ak Citinib (AG-13736), AZD-2171, CP-547632, IM-862, Macugen (pegaptanib), Nexavar (sorafenib, BAY43-9006), pazopanib (GW-786034), vataranib (PTK-787 or ZK-222584), Sutent (sunitinib or SU-11248), VEGF trap, Zactima (vandetanib or ZD-6474), and the like.

Antibiotics include aclarubicin, actinomycin D, amrubicin, annamycin, Adriamycin (doxorubicin), Blenoxane (bleomycin), daunorubicin, Caelyx and Myocet (liposomal doxorubicin), elsamitrucin , epirubicin, glarubicin, idarubicin, mitomycin C, nemorubicin, neocarzinostatin, peplomycin, pyrarubicin, lebecamycin, stimalamer, streptozocin, Valstar (valrubicin ), and intercalating antibiotics such as ginostatin.

Topoisomerase inhibitors include aclarubicin, 9-aminocamptothecin, amonapide, amsacrine, becatecarin, belotecan, BN-80915, Camptosar (irinotecan hydrotloride), gamptothecin , Cardioxane (dexrazoxane), diflomotecan, edotecarin, Ellence and Pharmorubicin (epirubicin), etoposide, exatecan, 10-hydroxylcamptothecin, jimatecan, rutotecan , mitoxantrone, oratecin, pyrabucin, picantrone, rubitecan, sobuic acid, SN-38, tafluposide, topotecan, and the like.

Antibodies include Avastin (bevacizumab), CD40-specific antibody, chTNT-1/B, denosumab, Erbitux (cetusimab), Humax-CD4 (zanolimumab), IGFlR-specific antibody, Lintuzumab, Panorex (edrecolomab), Rencarex (WX G250), Rituxan (rituximab), ticilimumab, trastuzumab, CD20 antibody types I and II, and the like.

Hormone Therapeutics: Arimidex (Anastrozole), Aromasin (Exemestane), Arzoxifen, Casodex (Bicalutamide), Cetrotide (Cetrotide), Degarelix, Deslorrelin, Desopan (Trillostane), Dexamethasone, Drogenil (Flulutamide), Evista (raloxifene), Afema (fadrozole), Fareston (toremifene), Faslodex (Fulvestrant), Femara (Retro) sol), formestane, glucocorticoids, Hectorol (docecalciferol), Renagel (sevelamer carbonate), lasofoxifen, leuprolide acetate, Megace (megestrol), Mifeprex (mifepristone), Nilandron (nilutamide), tamoxifen including Nolvadex (tamoxifen citrate), Plenaxis (abarelix), prednisone, Propecia (finasteride), rilostane, Suprefact (buserellin), luteinizing hormone (LHRH) including Trelstar (tryptorelin), histrelin including Vantas (histrelin implant), Modrastane (trillostane), Zoladex (goserelin), and the like.

Deltoids and retinoids include seocalcitol (EB1089 or CB1093), lexacalcitol (KH1060), fenretinide, tretinoin, including Panretin (aliretinoin), Atragen (liposomal tretinoin), Targretin ( bexarotene), LGD-1550, and the like.

PARP inhibitors include ABT-888, olaparib, KU-59436, AZD-2281, AG-014699, BSI-201, BGP-15, INO-1001, ONO-2231, and the like.

Plant alkaloids include vincristine, vinblastine, vindesine, vinorelbine, and the like.

Proteasome inhibitors include Velcade (bortezomib), MG132, NPI-0052, PR-171, and the like.

Examples of immune agents include interferon and other immune enhancing agents. Interferons include interferon alpha, interferon alpha-2a, interferon alpha-2b, interferon beta, interferon gamma-1a, Actimmune (interferon gamma-1b), interferon gamma-nl, combinations thereof, and the like. Other drugs include alphaperon (IFN-α), BAM-002 (oxidized glutathione), Beromun (tasonemin), Bexxar (tositumomab), Campath (alemtuzumab), CTLA4 (cytotoxic lymphocyte antigen 4) ), dacarbazine, denileukin, epratuzumab, Granocyte (lenograstim), lentinan, leukocyte alpha interferon, imiquimod, MDX-OlO (anti-CTLA-4), melanoma vaccine, mitumomab , molgramostim, Mylotarg (gemtuzumab ozogamicin), Neupogen (filgrastim), Onco VAC-CL, Ovarex (oregobumab), femtumomab (Y-muHMFGl), Provenge (cifulucel) -T), Sagaramostim, Cyzopyran, Theseleukin, Theracys (BCG or Bacillus Calmet-Gerin), Ubenimex, Virulizin (Immunotherapy, Lorus Pharmaceuticals), Z-100 (Maruyama specific substrate) or SSM), WF-10 (tetrachlorodecaoxide or TCDO), Proleukin (aldesleukin), Zadaxin (thymalfacin), Zenapax (daclizumab), Zevalin (90Y-ibritumab tiuxetane), and the like.

Biological response modifiers are agents that modify an organism's defense mechanism or biological responses such as survival, growth or differentiation of tissue cells to have anti-tumor activity, and include crestin, lentinan, sizofuran, fishivanil, PF-3512676 (CpG -8954), Ubenimex, and the like.

Pyrimidine analogs include cytarabine (cytosine arabinoside, ara C or arabinoside C), doxyfluridine, Fludara (fludarabine), 5-FU (5-fluororacil), floxuridine, Gemzar (gemcitabine), Tomudex (raltitrexd), triacetyluridine, Troxatyl (troxacitabine), and the like.

Purine analogs include Lanvis (thioguanine), Purinethol (mercaptopurine), and the like.

Antimitotic agents batavulin, epothilon D (KOS-862), N-(2-((4-hydroxy-phenyl)amino)pyridin-3-yl)-4-methoxybenzenesulfonamide, ixabe pilon (BMS-247550), paclitaxel, Taxotere (docetaxel), larotaxel (PNU-100940, RPR-109881 or XRP-9881), patupilone, vinflunin, ZK-EPO (synthetic epothylone), and the like. .

Ubiquitin ligase inhibitors include MDM2 inhibitors such as Nutlin, NEDD8 inhibitors such as MLN4924, and the like.

The composition of the present invention can be used as a radiation sensitizer to increase the effectiveness of radiation therapy. Examples of radiation therapy include, but are not limited to, external radiation therapy (XBRT), remote therapy, brachytherapy, sealed source radiation therapy, open source radiation therapy, and the like.

Additionally or alternatively, the compositions of the present invention may be administered in combination with one or more anti-tumor agents, or Abraxane (ABI-007), ABT-100 (farnesyl transferase inhibitor), Advexin (Ad5CMV-p53) Vaccine or Contusugene radenovec), Altocor or Mevacor (lovastatin), Ampligen (poly(I)-poly(C12U), synthetic RNA), Aptosyn (exisulind), Aredia (pamidronic acid), agrabine, L-asparaginase, atamestane (l-methyl-3,17-dione-androsta-l,4-diene), Avage (tazarotene), AVE-8062 (combretastatin derivatives) ), BEC2 (mitumomab), kakectin or kakeksin (tumor necrosis factor), Canvaxin (melanoma vaccine), CeaVac (cancer vaccine), Celeuk (celmolleukin), Ceplene (histamine dihydrochloride) containing histamine, Cervarix (AS04 adjuvant adsorbed human papilloma virus (HPV) vaccine), CHOP (Cytoxan (cyclophosphamide) + Adriamycin (doxorubicin) + Oncovin (vincristine) + prednisone), cumbretastatin A4P, Cypat (cyproterone), DAB(389)EGF (catalytic and translocation domain of diphtheria toxin fused via a human epidermal growth factor His-Ala linker), dacarbazine, dactinomycin, Dimericine (T4N5 liposome lotion), 5,6-dimethylsanthenone-4-acetic acid (DMXAA), discodemolide, DX-8951f (exatecan mesylate), enyluracil (ethynyluracil), Evizon (squalamine lactate) Squalamine, Enzastaurine, EPO-906 (Epotylone B), Gardasil (tetravalent human papilloma virus (Type 6, 11, 16, 18) recombinant vaccine), Gastrimmune, Genasense (Oblimersen) ), GMK (Garlioside Conjugate Vaccine), GV AX (Prostate Cancer Vaccine), Halofuginone, Hysterellin, Hydroxycarbamide, Ibandronic Acid, IGN-101, IL-13-PE38, IL-13-PE38QQR (Sintredechin vesodotox), IL-13-pseudomonas exotoxin, interferon-α, interferon-γ, Junovan and Mepact (miphamutide), lonapanib, 5,10-methylintetrahydrofolate, milte Forcin (hexadecyl-phosphocholine), Neovastat (AE-941), Neutrexin (trimetrexate glucuronate), Nipent (pentostatin), Onconase (lanpyrnase, ribonuclease enzyme) , Oncophage (Vitespen, melanoma vaccine treatment), OncoVAX (IL-2 vaccine), Orathecin (rubitecan), Osidem (antibody-based cell drug), Ovarex MAb (murine monoclonal antibody), paclitaxel albumin-stabilized nanoparticles, paclitaxel, Pandimex (aglycone saponin derived from ginseng, including 20(s)-protopanaxadiol (aPPD) and 20(s)-protopanaxatriol (aPPT)), panitumumab, Panvac-VF (experimental) cancer vaccine), pegaspargase, peginterferonalpa (PEG interferon A), phenoxodiol, procarbazine, levimastat, Removab (catumasomab), Revlimid (lenaridomide), RSR13 ( Epaproxiral), Somatuline LA (lanreotide), Soriatane (Acitretin), Staurosporine (Streptomyces staurospores), Tarabostet (PTlOO), Targretin (Bexarotene), Taxoprexin (Docco) sahexenoic acid (DHA) + paclitaxel), Telcyta (canphosphamide, TLK-286), Temodar (temozolomide), tesmilifene, tetrandrine, thalidomide, Theratope (STn-KLH vaccine) ), Thymitaq (noratrex dihydrochloride), TNFerade (adenovector: DNA carrier containing a gene for tumor necrosis factor-α), Tracleer or Zavesca (Bosentan), TransMID-107R (KSB-311, diphtheria toxin), Tretinoin (Retin-A), Trisenox (Arsenic Trioc) seed), Ukrain (alkaloid derivatives from celandine plants), Virulizin, Vitaxin (anti-αvβ3 antibody), Xcytrin (motexapine gadolinium), Xinlay (atrasentan), Xyotax (paclitaxel polyglomex) ), Yondelis (trabectedin), ZD-6126 (N-acetylcholcinol-O-phosphate), Zinecard (dexrazo acid), zoledronic acid, Zorobicin, etc. can be administered.

In one embodiment, a composition of the invention, e.g., a composition comprising ABT-263, comprises an anti-apoptotic Bcl-2 protein, an anti-apoptotic Bcl-X_L It is administered in a therapeutically effective amount to patients in need thereof to treat a disease caused by overexpression of one or more of the protein and the anti-apoptotic Bcl-w protein.

In another embodiment, a composition of the invention, e.g., a composition of the invention comprising ABT-263, is administered in a therapeutically effective amount to a subject in need thereof to treat a disease of abnormal cell growth and/or dysregulated apoptosis. is administered

Examples of such diseases include, but are not limited to, cancer, mesothelioma, bladder cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or intraocular melanoma, ovarian cancer, breast cancer, uterine cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, Vaginal carcinoma, vulvar carcinoma, bone cancer, colorectal cancer, rectal cancer, cancer of the anal region, stomach cancer, gastrointestinal (stomach, colon and/or duodenal) cancer, chronic lymphocytic leukemia, acute lymphocytic leukemia, esophageal cancer, small intestine cancer, endocrine system cancer , thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, testicular cancer, hepatocellular (liver and/or bile duct) cancer, primary or secondary central nervous system tumor, primary or secondary brain tumor, Hodgkin's disease , chronic or acute leukemia, chronic myelogenous leukemia, lymphocytic lymphoma, lymphoblastic leukemia, follicular lymphoma, lymphocytic malignancy of T-cell or B-cell origin, melanoma, multiple myeloma, oral cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, kidney cancer and/or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, neoplasms of the central nervous system, primary central nervous system lymphoma, non-Hodgkin's lymphoma, spinal axis tumor, brainstem glioma, pituitary adenoma, adrenal cortical cancer, gallbladder cancer, spleen cancer, cholangiocarcinoma, fibrosarcoma, neuroblastoma, retinoblastoma or a combination thereof .

In a more specific embodiment, a composition of the present invention, for example a composition of the present invention comprising ABT-263, is administered to bladder cancer, brain cancer, breast cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, acute lymphocytic leukemia, rectal cancer, esophageal cancer, Hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancy of T-cell or B-cell origin, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer or non It is administered in a therapeutically effective amount to a subject in need thereof to treat bowel cancer.

According to any of these embodiments, the composition is administered as a single regimen or in combination with one or more additional therapeutic agents.

For example, mesothelioma, bladder cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or intraocular melanoma, ovarian cancer, breast cancer, uterine cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, bone cancer, colon cancer , rectal cancer, cancer of the anal region, stomach cancer, gastrointestinal (stomach, colon and/or duodenal) cancer, chronic lymphocytic leukemia, acute lymphocytic leukemia, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue tissue sarcoma, urethral cancer, penile cancer, testicular cancer, hepatocellular (liver and/or bile duct) cancer, primary or secondary central nervous system tumor, primary or secondary brain tumor, Hodgkin's disease, chronic or acute leukemia, chronic myelogenous leukemia, Lymphocytic lymphoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancy of T-cell or B-cell origin, melanoma, multiple myeloma, oral cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, kidney cancer and/or ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, neoplasm of central nervous system, primary central nervous system lymphoma, non-Hodgkin's lymphoma, spinal axis tumor, A method of treating brainstem glioma, pituitary adenoma, adrenal cortical cancer, gallbladder cancer, spleen cancer, cholangiocarcinoma, fibrosarcoma, neuroblastoma, retinoblastoma, or a combination thereof comprises (a) a composition of the present invention, for example, ABT-263. administering to a subject a composition of the present invention comprising and (b) a therapeutically effective amount of one or more of etoposide, vincristine, CHOP, rituximab, rapamycin, R-CHOP, RCVP, DA-EPOCH-R, or bortezomib; includes

In a specific embodiment, a composition of the invention, e.g., a composition of the invention comprising ABT-263, is administered to a subject in need thereof for the treatment of a lymphocytic malignancy, such as B-cell lymphoma or non-Hodgkin's lymphoma. It is administered in a therapeutically effective amount in combination with a therapeutically effective amount of etoposide, vincristine, CHOP, rituximab, rapamycin, R-CHOP, RCVP, DA-EPOCH-R or bortezomib.

The present invention provides a method of maintaining a therapeutically effective plasma concentration of ABT-263 and/or one or more metabolites thereof in the bloodstream of a human cancer patient, said method comprising the subject ABT-263 or a pharmaceutically acceptable A nanoparticulate suspension comprising a salt, a prodrug, a salt of a prodrug or a metabolite thereof, in particular a crystalline salt of ABT-263, for example ABT-263 bis-UCL, is administered in a dose of about 50 to 500 mg ABT-263 per day. and administering at an average dosing interval of about 3 hours to 7 days at a dose equivalent to

What constitutes a therapeutically effective plasma concentration depends, inter alia, on the specific cancer the patient has, the stage of said cancer, the severity and aggressiveness of the cancer and the outcome sought (e.g., stabilization, reduction of tumor growth, tumor shrinkage, reduced risk of metastasis, etc.). ) depends on The plasma concentration should be sufficient to provide an advantage in terms of cancer treatment, but not excessive enough to cause unacceptable or intolerable adverse side effects.

For the treatment of lymphocytic malignancies such as cancer in general and in particular non-Hodgkin's lymphoma, plasma concentrations of ABT-263 should in most cases be maintained within the range of about 0.5 to about 10 μg/ml. Therefore, during the course of ABT-263 treatment, steady state C_maxshould generally not exceed about 10 μg/ml, steady state C_minshould generally not fall below about 0.5 μg/ml. Also within the ranges provided above, a C of about 5 or less, such as about 3 or less, at steady state._max/C_min It will be found desirable to select a daily dose and average dosing interval effective to provide a ratio. The longer the dosing interval, the more C_max/C_min It will be appreciated that the proportions tend to be larger. Illustratively, the method of the present invention comprises about 3 to about 8 μg/ml of ABT-263 C at steady state._max and about 1 to about 5 μg/ml C_mincan be aimed at

According to this embodiment, an effective daily dose to maintain therapeutically effective ABT-263 plasma levels is from about 50 to about 500 mg. In most cases, a suitable daily dose is from about 200 to about 400 mg. Illustratively, the daily dose may be, for example, about 50, about 100, about 150, about 200, about 250, about 300, about 350, about 400, about 450 or about 500 mg.

According to this embodiment, the average dosing interval effective to maintain therapeutically effective ABT-263 plasma levels is from about 3 hours to about 7 days. In most cases, suitable average dosing intervals are from about 8 hours to about 3 days, or from about 12 hours to about 2 days. A once-daily (qd) dosing regimen is often suitable.

With respect to this aspect, ABT-263 is illustratively ABT-263 <i>bis</i>-HCl or other crystalline ABT-263 salts are present in the pharmaceutical composition. Any ABT-263 composition of the present invention, as defined more fully above, may be used.

As in other embodiments, administration according to this embodiment may be with or without food, ie, non-fasted or on an empty stomach. In general, it is preferred that the compositions of the present invention be administered to the patient on a non-fasting basis.

Example

The following examples are illustrative only and are not limiting to the facts found in any way.

All ABT-263 amounts, including concentrations and doses, are expressed as doses equivalent to free base in the given examples, unless otherwise noted. When ABT-263 is used as the bis-HCl salt, 1.076 mg of ABT-263 bis-HCl provides the equivalent of 1 mg ABT-263 free base.

<u>Example</u><u> 1: Exemplary nanoparticulate form </u><u>in suspension</u><u> Produce</u>

ABT-263 nanoparticulate suspension formulations are prepared by high pressure homogenization as shown below. The formulation has the following composition in water (all percentages expressed in weight/volume).

<u>Formulation I (comparative)</u>

ABT-263 bis-HCl 5% (equivalent to 4.65% free base)

Poloxamer 188 3%

<u>formulation </u><u>II</u><u> (Example of the present invention)</u>

ABT-263 bis-HCl 5% (equivalent to 4.65% free base)

Poloxamer 188 3%

NaHCO₃ 8.4%

Prepare an aqueous solution containing the indicated amount of Poloxamer 188 (Pluronic F68) and, in the case of Formulation II, the indicated amount of sodium bicarbonate. A sufficient amount of crystalline ABT-263 bis-HCl to provide a 5% weight/volume (50 mg/ml) suspension was dispersed in each aqueous solution using a Sonifier homogenizer (Branson Ultrasonic, Danbury, CT). The resulting dispersion was added to the sample reservoir of a Microfluidizer M-11OL processor (Microfluidics International Corp., Newton, MA) and treated at 12,000 psi (approximately 82.5 MPa) for 2 hours. The sample temperature was maintained at a temperature of 20 ± 2 °C by dispersing through a heat exchanger immersed in a water bath connected to a cooling device.

The suspensions thus obtained (formulations I and II) were measured for particle size immediately after preparation and after storage at 5° C. for 14 days (see Example 2). Formulation II was used in an oral pharmacokinetic (PK) study in dogs (see Example 3).

<u>Example</u><u> 2: </u><u>of nanosuspension</u><u> Effect of sodium bicarbonate on particle size stability</u>

Formulations I and II are compared with respect to their particle size distribution (D₉₀ and D₅₀). Particle size measurements were performed immediately after preparation of the suspension (t=0) and after storage at 5° C. for 14 days. Particle size was also measured at t=0 for the suspensions after dilution of 1 ml of each suspension in 20 ml 0.9% sodium chloride solution (NaCl). Data are presented in Table 1.

Table 1 D of Nanosuspension Formulations I and II₉₀ and D₅₀Particle size (μm)

<img file="KR20120052937A_D0017.tif" />

<u>Example</u><u> 3: Exemplary nano </u><u>in suspension</u><u></u><u>Pharmacokinetics</u>

The single-dose pharmacokinetics of Formulation II of Example 1 were measured after oral administration of 5 mg/kg to non-fasting beagle dogs (n=4). The formulation was administered by two methods: oral gavage and capsules. Formulation II was administered only by oral gavage to histamine-pretreated fasting dogs (n=4). ABT-263 bis-HCl in lipid medium (formulation C, prepared from ABT-263 bis-HCl powder dissolved at a concentration of 25 mg/ml in a 90:10 mixture of Formulation C, Phosal 53 MCT and ethanol) for comparative purposes. of the solution formulation was administered to non-fasting dogs. Formulation C was used to evaluate ABT-263 in clinical studies. Phosal 53 MCT is a proprietary blend provided by Phospholipids Limited and contains 53% phosphatidylcholine and 29% medium chain triglycerides.

Serial heparinized blood samples were obtained from the jugular vein of each animal immediately before dosing and 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 9, 12, 15 and 24 hours after dosing. Plasma was separated by centrifugation (2000 rpm for 10 min at approximately 4° C.) and ABT-263 was isolated using protein precipitation with acetonitrile.

ABT-263 and internal standard were carried out from each other and on a 50 x 3 mm Keystone Betasil CN 5 μm column with an acetonitrile/0.1% trifluoroacetic acid mobile phase (volume ratio of 50/50) from each other and at a flow rate of 0.7 ml/min. was separated from the co-extracted contaminants. Analyze on a biomolecular mass spectrometer Sciex API3000 with a heated nebulizer footprint. ABT-263 and internal standard peak areas were determined using Sciex MacQuan software. The plasma drug concentration of each sample was calculated by least squares linear regression (unweighted) method of the peak area ratio of the spike plasma standard to concentration (upper/internal standard). Plasma concentration data were applied to multi-exponential curve fitting using WinNonlin 3 (Pharsight).

The area under the plasma concentration-time curve from 0 to t hours post-dose (time of last measured plasma concentration, here 24 hours) was calculated using the linear trapezoidal rule for plasma concentration-time profiles.

Mean plasma concentrations over 24 hours post-dose are shown in FIG. 1 .

The calculated mean values of the PK variables are summarized in Table 2.

PK Variables (Mean ± SEM) in Table 2 (non-fasting unless otherwise noted)

<img file="KR20120052937A_D0018.tif" />