HK1144669B - Crystalline form of (e) 4-[[4-[[4-(2-cyanoethenyl)-2,6-dimethylphenyl]amino]-2 pyrimidinyl]amino]benzonitrile - Google Patents

Description

(E) Crystal forms of 4- [ [4- [ [4- (2-cyanoethenyl) -2, 6-dimethylphenyl ] amino ] -2-pyrimidinyl ] amino ] benzonitrile

Technical Field

The present invention relates to crystalline forms of TMC278, uses and methods of preparation thereof. The invention also relates to pharmaceutical formulations comprising such crystalline forms.

Background

The treatment of Human Immunodeficiency Virus (HIV) infection, known as the cause of acquired immunodeficiency syndrome (AIDS), has been a major medical challenge. Currently available drug therapies include Nucleoside Reverse Transcriptase Inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), nucleotide reverse transcriptase inhibitors (NtRTIs), HIV-Protease Inhibitors (PIs), fusion inhibitors and more recently CCR5 and integrase inhibitors.

Although effective in inhibiting HIV, these drugs are confronted with the emergence of drug resistant mutants when used alone. This has led to the introduction of several combination therapies of anti-HIV drugs, often with different activity profiles. In particular, the introduction of "HAART" (highly active anti-retroviral therapy) has resulted in significant improvements in anti-HIV therapy, which has resulted in significant reductions in HIV-related morbidity and mortality. However, no combination therapy is currently available to completely eradicate HIV. Even HAART faces the problem of developing resistance, often due to non-compliance with the prescribed therapy. In these cases, HAART can be made effective again by replacing one of the components with one of the other. HAART combination therapy, if applied correctly, can suppress the virus to a level that may no longer cause an AIDS outbreak for many years, even decades.

One class of HIV drugs commonly used in HAART are the drugs of the NNRTIs, many of which are currently on the market, while several others are in various stages of development. One NNRTI currently under development is the compound 4- [ [4- [ [4- (2-cyanoethenyl) -2, 6-dimethylphenyl ] amino ] -2-pyrimidinyl ] amino ] benzonitrile, commonly known as rilpivirine, also known as R278474 or TMC 278. The compound not only shows remarkable activity against wild-type HIV, but also shows inhibitory activity against a variety of mutants thereof. The compound TMC278, its pharmacological activity and a series of methods for its preparation are described in WO 03/16306. The TMC278 described in this document is in a crystalline form, and this form is hereinafter referred to as "polymorph II" of TMC 278. The present invention relates to another polymorph of TMC278, which is hereinafter referred to as "polymorph I" of TMC 278. Polymorph I of TMC278, which has not been described previously, is a crystalline form with beneficial properties as will be outlined below.

With respect to activity, pharmacokinetic properties play an important role in the efficacy of any given drug. This, in turn, is consistent with the bioavailability of the drug, which affects the dosage required to achieve a therapeutically effective concentration of the drug in the patient. Drugs with low bioavailability require high dose administration, thereby increasing the risk of side effects. High doses also involve larger dosage forms or increased frequency of administration, or both. These factors can affect adhesion and concurrent effects of anti-HIV therapy. Inadequate treatment in turn increases the risk of emergence of mutant HIV strains.

TMC278 has a relatively low solubility in water, making bioavailability poor. Unexpectedly, it has now been found that the new crystalline forms of TMC278 have an increased intrinsic dissolution rate and a higher solubility in acidic aqueous media. These properties would be beneficial in terms of bioavailability to prepare new crystalline forms that are attractive for application in solid dosage forms, but may also be useful in certain liquid dosage forms, such as aqueous dispersions. The latter can be used in formulations for parenteral administration.

In addition, the use of a particular polymorphic form of an active ingredient may be recommended because the composition of the polymorphic mixture may vary from batch to batch, or may vary over time, thereby causing a change in the ratio of the active ingredients. If the polymorph does not remain constant in clinical and stability studies, the exact dose used or determined may not be comparable between batches. Once the pharmaceutical compound produced is administered to a human, it is important to identify the polymorphic form delivered in each dosage form, thereby ensuring that the same form is used in the manufacturing process and that the same amount of drug is included in each dose. Thus, it must be ensured that a single polymorph or a polymorph of some known composition is given.

The anti-HIV drugs currently in use require frequent relatively high dose administration. The number and/or volume of dosage forms to be administered is commonly referred to as "drug loaded negative". High negative drug loading prevents patients from taking the drug according to the prescribed dosage regimen, thereby not only reducing the therapeutic efficacy, but also causing the emergence of drug-resistant mutants. There is therefore a need for an anti-HIV therapy that avoids high negative drug loading, which involves the administration of relatively small amounts of dosage forms, and which does not require frequent administration. It would also be desirable to provide anti-HIV therapy that can be administered at long time intervals, such as one week, one month, or longer.

Current therapies fail to completely eradicate HIV, creating a continuing risk for HIV-infected patients to infect others. After the initial infection, a long time passes before the first symptoms of AIDS develop. Infected persons can live for many years without experiencing any AIDS effect and are therefore unaware of the risk of further transmission of the virus to other persons. Because individuals who come into contact with HIV-infected patients with infectious disease are extremely dangerous, it is important to prevent HIV infection. Especially those providing medical care to infected patients, such as physicians, nurses or dentists. Another group of at risk individuals are breastfed infants whose mothers are infected or at risk of infection, especially in developing countries where the alternative means of breastfeeding is less obvious. Therefore, there is a need for an easy to apply method that provides effective protection against HIV infection. It is another object of the present invention to provide such a prophylactic method.

The crystalline forms of TMC278 of the present invention may be formulated as micro-or nanoparticles as depot (depot) formulations for the treatment of HIV infection and the prevention of HIV infection. Nanoparticulate drugs are known, for example, from EP-A-O499299. It has been found that micro-or nanoparticle formulations of polymorph I of TMC278 can be administered intermittently at time intervals of one week or more such that the blood plasma levels are sufficient to inhibit the proliferation of HIV. Thereby reducing the number of administrations, which is beneficial for drug loading and patient compliance with medication. These micro-or nanoparticle formulations find use in the long-term treatment or prevention of HIV.

Polymorph I of TMC278 is particularly suitable for micro-or nanoparticle formulations due to its beneficial properties. These formulations are expected to produce higher blood plasma levels for a given amount of TMC278 administered. In addition, the desired plasma levels will be reached more quickly. When high safety margins are required, relatively high plasma levels may be required.

Description of the drawings

FIG. 1: KBr disperse phase IR Spectroscopy of TMC278 polymorph I

FIG. 2: x-ray powder diffraction (XRD) Pattern of polymorph I of TMC278

FIG. 3: differential Scanning Calorimetry (DSC): TMC278 polymorph I

FIG. 4: dissolution Profile of TMC278 polymorph I

FIG. 5: KBr disperse phase Infrared Spectroscopy of TMC278 polymorph II

FIG. 6: powder XRD Pattern of polymorph II of TMC278

FIG. 7: DSC: TMC278 polymorph form II

FIG. 8: dissolution Profile of TMC278 polymorph II

Summary of The Invention

The present invention relates to a particular crystalline form of TMC278, polymorph I of (E)4- [ [4- [ [4- (2-cyanoethenyl) -2, 6-dimethylphenyl ] amino ] -2-pyrimidinyl ] amino ] benzonitrile (i.e. TMC278), also known as rilpivirine. This polymorph is characterized by the biochemical parameters mentioned in the examples.

The invention also relates to a method for preparing the crystal form. In another aspect, the present invention relates to solid pharmaceutical compositions comprising polymorph I of TMC278 as active ingredient and a pharmaceutically acceptable carrier.

The present invention also relates to a pharmaceutical composition of micro-or nanoparticles for administration by intramuscular or subcutaneous injection comprising a therapeutically effective amount of polymorph I of TMC278 in the form of micro-or nanoparticles having a surface modifier adsorbed on the surface thereof, suspended in a pharmaceutically acceptable aqueous carrier.

The present invention also relates to a method of treating a patient infected with HIV, which method comprises administering to said patient by intramuscular or subcutaneous injection an anti-HIV effective amount of a micro-or nanoparticle composition as defined herein. In another aspect, the method is for long term treatment and the composition may be administered or administered intermittently at time intervals ranging from one week to one year, or one week to two years, or one month to three months. Alternatively, the invention relates to the use of micro-or nanoparticles as defined herein for the manufacture of a medicament for the treatment of HIV infection. Alternatively, the use is for long term treatment and the composition may be administered or administered intermittently at intervals ranging from one week to one year, or from one week to two years, or from one month to three months.

The present invention also relates to a method for preventing HIV infection in a patient at risk of HIV infection, which method comprises administering to said patient by intramuscular or subcutaneous injection an HIV infection preventing effective amount of a micro-or nanoparticle composition as defined herein. In another aspect, the method is for long term treatment and the composition may be administered or administered intermittently at intervals ranging from one week to one year, or one week to two years, or one month to three months. Alternatively, the invention relates to the use of micro-or nanoparticles as defined herein for the manufacture of a medicament for the prevention of HIV infection in a patient at risk of HIV infection. Alternatively, the use is for long term treatment and the composition may be administered or administered intermittently at intervals ranging from one week to one year, or from one week to two years, or from one month to three months.

Pharmaceutical compositions, methods of treatment or prophylaxis and use for the preparation of medicaments based on these compositions are described further below and are intended to form part of the present invention.

Detailed Description

As used herein, polymorphic forms of a compound refer to the same chemical entity, but differ in the arrangement of the crystalline forms.

Form I of TMC278 according to the present invention may also be referred to as "form I TMC 278", "polymorph I of TMC 278" or similar expressions, characterized by the physicochemical parameters listed below. The second polymorphic form of TMC278, referred to herein as form II, is a form obtained when the compound is synthesized using the procedures of WO 03/16306. It may also be referred to as "form II TMC 278", "polymorph II of TMC 278", or similar terms.

The chemical structure of TMC278 is represented by the formula:

TMC278 has two stereochemical configurations at the double bond of the cyanovinyl moiety, namely the E (Entgegen) configuration (E-isomer) and the Z (Zusammen) configuration (Z-isomer). Unless otherwise indicated, the term TMC278 or similar terms refer to the E-isomer, especially the E-isomer substantially free of the Z-isomer. Whenever reference is made herein to the E-isomer, this is meant to refer to the pure E-isomer or any isomeric mixture wherein the E-isomer is predominantly the E-and Z-isomers, i.e. isomeric mixtures containing more than 70%, or especially more than 80%, and more especially more than 90% of the E-isomer. Of particular importance is the E-isomer which is essentially free of the Z-isomer. "essentially free" in this context means E-Z-mixtures which are free or almost free of Z-isomer, such as isomer mixtures which comprise 90%, in particular 95%, or even 98% or 99% of E-isomer.

TMC278 is an HIV inhibitor, especially HIV-1, a pathogen-directed drug of human acquired immunodeficiency syndrome (AIDS). TMC278 is active against drug-resistant and multidrug-resistant HIV strains, particularly HIV strains that have inhibited acquisition of resistance by one or more non-nucleoside reverse transcriptase inhibitors (especially efavirenz, nevirapine, and delavirdine). TMC278 may find use in the treatment of other conditions associated with HIV infection, including thrombocytopenia, kaposi's sarcoma and infections of the central nervous system characterized by progressive demyelination, which leads to dementia and symptoms such as progressive dysphonia, dyskinesia and disorientation, peripheral neuropathy, Progressive Generalized Lymphadenopathy (PGL) and AIDS-related complex disease (ARC).

Polymorph I of TMC278 object of the present invention is useful for the treatment of HIV infected individuals and for the prevention of HIV infection. It can also be used for the treatment and prevention of the above mentioned HIV-related disorders. Accordingly, the present invention also relates to a method of treating, or preventing, an HIV infection in a human, comprising administering to a human in need thereof a therapeutically effective amount of polymorph I of TMC 278. Alternatively, the present invention relates to a method for the treatment of a disease associated with HIV infection in a human comprising administering to a human in need thereof a therapeutically effective amount of polymorph I of TMC 278.

Polymorph I of TMC278 exhibits increased solubility at lower pH and increased intrinsic dissolution rate compared to polymorph II. Due to the poor solubility of TMC278 these properties will have a beneficial effect on the bioavailability, so that effective blood plasma levels are more readily obtained, giving the active ingredient a better performance with respect to its antiviral effect. Thus, polymorph I of TMC278 is better absorbed in a more strongly acidic medium, such as gastric fluid.

Polymorph I of TMC278 can be prepared by dissolving TMC278 in a ketone solvent, such as di-C_1-4Alkyl ketones, such as 2-butanone, methyl isopropyl ketone, methyl isobutyl ketone, 2-pentanone, 3-pentanone and especially acetone, by heating to reflux temperature and cooling the solution, especially to below 0 ℃And (4) preparing. After removal of the solvent, crystals were obtained, which were dried. The starting material TMC278 may be prepared as described in WO 03/16306, term C_1-4Alkyl refers to straight or branched chain saturated hydrocarbons of 1 to 4 carbon atoms.

The present invention also relates to a solid pharmaceutical composition comprising a pharmaceutically acceptable carrier and as active ingredient a therapeutically effective amount of polymorph I of TMC 278. The composition may be in the form of a solid dosage form, such as a tablet or capsule, or a suspension. The pharmaceutical compositions of the present invention may be prepared by intimately mixing an effective amount of polymorph I of TMC278 as the active ingredient with a pharmaceutically acceptable carrier. The pharmaceutical compositions are preferably formulated in unit dosage forms suitable for oral administration, such as tablets and capsules. The carrier may include any of the usual pharmaceutical media such as wetting agents, fillers, diluents, glidants, binders, disintegrants, lubricants and optional excipients such as flavoring agents, sweeteners and colorants. The tablets may optionally be film coated. Suspensions for injection can also be prepared by using appropriate liquid carriers, suspensions, and the like.

The pharmaceutical composition in unit dosage form may comprise an amount of polymorph I of TMC278 in the range of about 5 to about 500mg, alternatively about 10mg to about 250mg, alternatively about 20mg to about 200mg, alternatively about 25mg to about 150mg, i.e. about 25mg, about 50mg, about 75mg, about 100mg or about 150 mg. The pharmaceutical compositions described herein are of importance and comprise:

(a) 5-50% polymorph I of TMC 278;

(b) 0.01-5% of a humectant;

(c) 40-92% of a diluent;

(d) 0-10% of a polymer;

(e) 2-10% of a disintegrant;

(f) 0.1-5% of a glidant;

(g) 0.1-1.5% of lubricant.

Furthermore, polymorph I of TMC278 can be converted into micro-or nanoparticle suspensions, which can be used for long-term treatment and long-term prevention of HIV infection, requiring only a limited number of administrations. This is beneficial in terms of loading the medication and patient compliance with the prescribed dosage regimen.

The term "preventing HIV infection" relates to preventing or avoiding infection of a patient with HIV. The source of infection may vary, such as HIV-containing materials, especially HIV-containing body fluids such as blood or semen or other HIV-infected patients. Prevention of HIV infection involves preventing the transmission of the virus from HIV-containing materials or from HIV-infected individuals to uninfected persons, or preventing the entry of the virus into uninfected persons. Transmission of HIV can be by transmission of HIV for any known reason, such as by sexual transmission or by contact with the blood of an infected patient, such as when providing care to medical personnel of the infected patient to handle blood samples or transfusions. Transmission can also be by contact with infected cells, such as when laboratory tests are performed with HIV infected cells.

The term "treating an HIV infection" as used herein relates to treating a patient infected with HIV. The term "treating an HIV infection" also relates to the treatment of diseases associated with the above mentioned HIV infection. The terms "treatment of HIV infection", "anti-HIV therapy" and similar terms refer to therapy that reduces the viral load of HIV (expressed as the number of viral RNA copies in a particular volume of serum). The more effective the treatment, the lower the viral load. The amount of virus should preferably be reduced as low as possible, for example below about 200 copies (copies)/ml, especially below about 100 copies/ml, more especially below about 50 copies/ml, if possible below the detection limit of the virus. A 1, 2 or even 3 order reduction in viral load (e.g., a reduction of about 10 to about 10)²Or even more, such as about 10³Of the order of magnitude) is an indication that the treatment is effective. Another parameter for measuring the effectiveness of anti-HIV therapy is the CD4 count, which in normal adults ranges from 500-1500 cells per μ l. Decreased CD4 counts are indicative of HIV infection, and AIDS may develop if there is a decrease of about 200 cells per μ l. Increased CD4 counts, e.g., about 50, 100, 200 or more cells per μ l, are also anti-Indications for the effectiveness of HIV treatment. In particular, the CD4 count should be increased to a level above about 200 cells/. mu.l or above about 350 cells/. mu.l. The extent of HIV infection can be diagnosed with viral load or CD4 counts or both.

The term "effective in treating HIV" and similar terms refer to therapies that reduce viral load or increase CD4 counts, or both, as described above. The term "effective against HIV" and similar terms refer to the situation when the relative number of newly infected patients in a population exposed to a source of HIV infection (e.g., HIV-containing material or HIV-infected patients) is reduced. Effective prophylaxis can be determined, for example, by determining a mixed population of HIV-infected and non-infected individuals, i.e., whether the relative number of newly infected individuals decreases when comparing non-infected individuals treated with the micro-or nanoparticle compositions of the present invention to untreated non-infected individuals. This decrease can be measured by statistical analysis of the number of infected and non-infected individuals in a given population over a period of time.

The terms "therapeutically effective amount", "an amount effective in preventing HIV infection" and similar terms refer to the amount of the active ingredient TMC278 that produces effective blood plasma levels. By "effective plasma levels" is meant blood plasma levels of the HIV inhibitor TMC278 that provide an effective treatment or effective prevention of HIV infection. The term "patient" especially refers to a human.

The term "micro-or nanoparticles" refers to particles in the micro-or nano-range. In one embodiment, the micro-or nanoparticle compositions of the present invention comprise polymorph I TMC278 in the form of nanoparticles. The average effective particle size of the micro-or nanoparticles of the present invention may be below about 50 μm, or below about 20 μm, or below about 10 μm, or below about 1000nm, or below about 500nm, or below about 400nm, or below about 300nm, or below about 200 nm. The lower limit of the average effective particle size may be as low as, for example, about 100nm or as low as about 50 nm. In one embodiment, the average effective particle size is in the range of from about 50nm to about 50 μm, alternatively from about 50nm to about 20 μm, alternatively from about 50nm to about 10 μm, alternatively from about 50nm to about 1000nm, alternatively from about 50nm to about 500nm, alternatively from about 50nm to about 300nm, alternatively from about 100nm to about 250nm, alternatively from about 125nm to about 175nm, for example about 130nm or about 150 nm.

The term average effective particle size as used herein has its conventional meaning and can be determined by particle size determination techniques known in the art, such as sedimentation field flow fractionation, photon correlation spectroscopy, laser diffraction or disk centrifugation. The average effective particle size referred to herein may be related to the volume distribution of the particles. In this case, "an average effective particle size of less than about 50 μm" means that at least 50% by volume of the particles have a particle size of less than the effective average 50 μm, and the same applies to the other effective particle sizes mentioned. In a similar process, the average effective particle size may be related to the weight distribution of the particles, but it generally yields the same or about the same value as the average effective particle size.

Micro-or nanoparticle compositions of polymorph I of TMC278 provide release of the active ingredient TMC278 over an extended period of time, and they may therefore also be referred to as slow-release or delayed-release compositions, or depot formulations. After administration, these compositions remain in the body and stably release TMC278, retaining the active ingredient in the patient's body for an extended period of time, thereby providing an anti-HIV therapeutic effect or an effect of preventing HIV infection. Once administered, the plasma levels of TMC278 are relatively stable (more or less stable), i.e. they fluctuate within a defined range. This plasma level is close to a relatively stable peptide pattern or close to a zero order release rate. In some cases, there may be an initial plasma concentration peak after administration.

The term "extended period" as used herein refers to a period (or time period) which may be from one week to one year or up to two years, or a period in the range of 1-2 weeks, or 2-3 weeks, or 3-4 weeks, or a period in the range of 1-2 months, or 2-3 months, or 3-4 months, or 3-6 months, or 6-12 months, or 12-24 months.

The blood plasma level of the active ingredient TMC278 should be above a threshold value. In the case of a therapeutic application, the threshold is the minimum blood plasma level at which TMC278 provides an effective treatment for HIV infection. In the case of prevention of HIV infection, the threshold is the minimum blood plasma level at which TMC278 is effective in preventing transmission of HIV infection.

"Long-term" or similar terms as used in reference to terms such as "long-term prevention of HIV infection" or "long-term treatment of HIV infection" refer to a period that can range from one week to one year or up to two years or more (e.g., 5 or 10 years). Especially in the case of treatment of HIV infections, these periods will be longer, as dictated by one to several years. These periods may also be relatively short, especially in the case of prophylaxis, such as one week to one year.

Micro-or nanoparticle compositions of polymorph I of TMC278 may be administered at different time intervals. When used to prevent HIV infection, the micro-or nanoparticle compositions of the present invention may be administered only once or a limited number of times, such as 2, 3, 4, 5, or 6, or more. When a prophylactic effect is desired, administration over a limited period of time, such as during periods of risk of infection, may be recommended.

The micro-or nanoparticle compositions of the present invention may be administered at the time intervals mentioned above, such as a time interval in the range of one week to one month, or in the range of one month to three months, or in the range of three months to six months, or in the range of six months to twelve months, for example 1 time every 2 weeks, or 1 time per month or 1 time every 3 months. In another embodiment, the time interval is in the range of 1 to 2 weeks, alternatively 2 to 3 weeks or 3 to 4 weeks, alternatively 1 to 2 months, alternatively 2 to 3 months, alternatively 3 to 4 months, alternatively 3 to 6 months, alternatively 6 to 12 months, alternatively 12 to 24 months. The time interval may be at least one week, but may also be several weeks, such as 2, 3, 4, 5 or 6 weeks, or one month, or 2, 3, 4, 5 or 6 months or longer, such as 9 or 12 months. To further improve compliance, patients may be instructed to take medication on a day of the week, wherein the composition is administered on a weekly schedule, or on a fixed day of each month of a monthly schedule.

The length of the time interval between each administration of the micro-or nanoparticle compositions of the invention may vary. For example, the time interval may be selected as a function of plasma levels. The interval may be shortened when the blood plasma level of TMC278 is considered too low, or the interval may be lengthened when the blood plasma level of TMC278 is considered too high. The micro-or nanoparticle compositions of the present invention may be administered at equal intervals without any additional administration in between. Intervals of the same length have the advantage of a simple administration protocol, such as administration on the same day of the week, or on the same day of the month.

The dose (or amount) of TMC278 administered depends on the amount of TMC278 in the micro-or nanoparticle compositions of the invention or the amount of a given composition administered. When higher blood plasma levels are required, one or two higher concentrations of the combination of TMC278 or more of a given composition may be administered. If a lower plasma level is required and vice versa. Combinations of different time intervals and different dosages may also be selected to achieve certain desired plasma levels. The dose (or amount) of TMC278 administered will also depend on the frequency of administration (i.e. the time interval between administrations). Generally, the dosage is higher when the frequency of administration is lower.

Dosage regimens are also contemplated depending on whether HIV infection is prevented or treated. In the therapeutic context, the dose or frequency of administration or both of TMC278 administered may be selected such that the blood plasma concentration of TMC278 is kept at a minimum blood plasma level (or C)_min) Above. The latter term refers to providing plasma levels of TMC278 that are effective in the treatment of HIV. In particular, the plasma levels of TMC278 are kept at a level above a minimum plasma level, i.e. above about 10ng/ml, or above about 13.5ng/ml, or above about 15ng/ml, or above about 20ng/ml, or above about 40ng/ml or higher, e.g. above about 50ng/ml, or above about 90ng/ml, or above about 270ng/ml or above about 540 ng/ml. Alternatively, the blood plasma levels of TMC278 may be kept in a range, in particular a range starting from a minimum blood plasma level selected from those mentioned above to a higher blood plasma level selected from those mentioned above, and selected from 500ng/ml and 1000ng/ml, e.g. from about 10 to about 1020. From about 20 to about 90, from about 90 to about 270, from about 270 to about 540, from about 540 to about 1000, each time from the approximate value specified (ng/ml) to from the approximate value specified (ng/ml).

In the case of HIV prevention, the term "minimum plasma level" (or C)_min) Means that the lowest blood plasma level of TMC278 that provides effective prevention of HIV infection is provided. For the prevention of HIV, the plasma levels of TMC278 may be kept at a level above the minimum blood plasma level mentioned above in connection with treatment, or may be kept at a lower level, such as at a level above about 4ng/ml, or about 5ng/ml or about 8 ng/ml. The blood plasma levels of TMC278 can be kept at higher levels within safe limits. These higher levels start at about 50ng/ml or more. The plasma levels of TMC278 may be kept at levels within the above-mentioned ranges relevant for treatment, but lower limits thereof include plasma levels of about 4ng/ml or about 5ng/ml or about 8 ng/ml.

An advantage of TMC278 is that its relatively high blood plasma levels can be used without any significant side effects. Maximum plasma concentration (C) of TMC278_max) Relatively high levels may be achieved, even up to about 500ng/ml or 1000 ng/ml. In one embodiment, the amount and frequency of administration of TMC278 that is administered is selected such that the blood plasma concentration is maintained at a level, including at the maximum blood plasma level (or C as indicated above), over an extended period of time_max) And minimum plasma level (or C as indicated above)_min) In the meantime.

In certain cases it may be desirable to keep the blood plasma levels of TMC278 at relatively low levels, for example as close as possible to the minimum blood plasma levels referred to herein. This would require a reduction in the frequency of administration and/or the amount of TMC278 administered per administration. In case of a prophylactic effect, the blood plasma levels of TMC278 may be kept at relatively low levels. In other cases it may be desirable to keep the blood plasma levels of TMC278 at relatively high levels, e.g. the minimum blood plasma level may be equal to the minimum blood plasma level of TMC278 that provides an effective treatment of HIV, as mentioned herein for each particular level.

In the case of prophylaxis, the dose administered should be calculated based on the following quantities: from about 0.2 mg/day to about 50 mg/day, or from about 0.5 mg/day to about 50 mg/day, or from about 1 mg/day to about 10 mg/day, or from about 2 mg/day to about 5 mg/day, for example about 3 mg/day. To calculate the weekly dose, these amounts should be multiplied by 7, and for the monthly dose, these amounts should be multiplied by 30. Dosages for other dosing regimens are readily calculated by multiplying the daily dose by the number of days between doses. For treatment, the dose should be slightly higher and should be calculated based on the following amounts: from about 1 mg/day to about 150 mg/day, or from about 2 mg/day to about 100 mg/day, or from about 5 mg/day to about 50 mg/day, or from about 10 mg/day to about 25 mg/day, for example about 15 mg/day. The corresponding weekly or monthly doses may be calculated as mentioned above. For prophylactic use, the same dosage can be used for therapeutic use.

In one embodiment, the micro-or nanoparticles in the compositions of the present invention predominantly comprise polymorph I of TMC278 and the surface-modifying agent, the combined amount of which may comprise at least about 50%, or at least about 80%, or at least about 90%, or at least about 95% or at least about 99% of the micro-or nanoparticles.

The inventive micro-or nanoparticles have surface-modifying agents adsorbed on their surface, which act as wetting agents and stabilizers for suspensions.

Suitable surface-modifying agents may be selected from a variety of excipients, such as gelatin, casein, lecithin, salts of negatively charged phospholipids or acid forms thereof (e.g. phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, phosphoric acid, and salts thereof, such as alkali metal salts, e.g. sodium salts thereof, e.g. sodium egg phosphatidylglycerol, e.g. under the trade mark Lipoid^TMProducts available from EPG), gum arabic, stearic acid, benzalkonium chloride, polyoxyethylene alkyl ethers, such as polyethylene glycol ethers, e.g. cetomacrogol 1000, polyoxyethylene castor oil derivatives; polyoxyethylene stearate, colloidal silicon dioxide, sodium lauryl sulfate, sodium carboxymethylcellulose, bile acid salts such as sodium taurocholate, sodium deoxytaurocholate, sodium deoxycholate; methyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, aluminumMagnesium silicate (magnesium aluminate silicate), polyvinyl alcohol (PVA), poloxamers, such as Pluronic^TMF68, F108 and F127, which are block copolymers of ethylene oxide and propylene oxide; tetrabutyl phenol; vitamin E-TGPS (alpha-tocopheryl polyethylene glycol succinate, especially alpha-tocopheryl polyethylene glycol 1000 succinate); poloxamers, e.g. Tetronic^TM908(T908), which is a tetrafunctional block copolymer derived from the continuous addition of ethylene oxide and propylene oxide to ethylenediamine; (ii) a glucan; lecithin; dioctyl esters of sodium sulfosuccinic acid, e.g. under the trade mark Aerosol OT^TM(AOT) products sold; sodium dodecyl sulfate (Duponol)^TMP); under the trademark Triton^TMAn alkylaryl polyether sulfonate obtainable from X-200; polyoxyethylene sorbitan fatty acid ester (Tweens)^TM)20, 40, 60 and 80); sorbitan esters of fatty acids (Span)^TM20. 40, 60 and 80 or Arlacel^TM20. 40, 60 and 80); polyethylene glycol (e.g. under the trade mark Carbowax)^TM3550 and 934 sold); mixtures of sucrose stearate and sucrose distearate, e.g. under the trade mark Crodesta^TMF110 or Crodesta^TMProducts available as SL-40; cetyltrimethylammonium chloride (CTAC); polyvinylpyrrolidone (PVP). Two or more surface modifiers may be used in combination, if desired.

Particular surface modifying agents are selected from the group consisting of poloxamers, alpha-tocopherol polyethylene glycol succinate, polyoxyethylene sorbitan fatty acid esters and salts of negatively charged phospholipids or acid forms thereof. More particularly the surface modifier is selected from Pluronic^TMF108, vitamin E TGPS, Tweens^TM80 and Lipoid^TMAn EPG. One or more of these surface modifiers may be used. Pluronic^TMF108 corresponds to poloxamer 338 and is a polyoxyethylene/polyoxypropylene block copolymer, generally conforming to the formula HO- [ CH ]₂CH₂O]_x-[CH(CH₃)CH₂O]_y-[CH₂CH₂O]_Z-H, wherein the average values of x, y and z are 128, 54 and 128, respectively. Poloxamer 338 under the other trade name Hodag Nonionic^TM1108-F and Synperonic^TMPE/F108. In one embodiment, the surface modifying agent comprises a combination of a polyoxyethylene sorbitan fatty acid ester and a phosphatidylglycerol salt (especially an egg phosphatidylglycerol salt).

The relative amounts (w/w) of polymorph I of TMC278 and surface modifier may vary, but may be in the range of from 1: 2 to about 20: 1, especially in the range of from 1: 1 to about 10: 1, such as about 4: 1.

Micro-or nanoparticles of polymorph I of TMC278 can be prepared by mechanical means via particle size reduction (nanocrystallization). Polymorph I of TMC278, preferably in micronized form, is produced in an aqueous dispersion medium in the presence of a surface modifier and a grinding medium is applied to reduce the particle size to the required effective particle size. The general procedure for preparing the particles of the invention comprises:

(a) obtaining polymorph I of TMC278 in micronized form;

(b) adding micronized TMC278 to a liquid medium to form a premix/predispersion; and

(c) reducing the premix to an average effective particle size with a mechanical tool in the presence of grinding media.

Polymorph I of TMC278 in micronized form can be prepared using techniques known in the art. The average effective particle size of the TMC278 active agent in the pre-dispersion may be less than about 100 μm, as determined by sieve analysis. When the average effective particle size is greater than about 100 μm, it is preferably reduced to below about 100 μm. Micronized polymorph I of TMC278 is then added to an aqueous medium to form a pre-dispersion.

Mechanical means for reducing the effective average particle size of polymorph I of TMC278 include roller mills or similar approaches using beads, e.g. ZrO₂Beads. The volume reduction is carried out at a temperature at which the TMC278 compound is not significantly degraded, preferably less than 30-40 ℃, e.g. at room temperature, if necessary with cooling.

The micro-or nanoparticle compositions of the present invention comprise a pharmaceutically acceptable aqueous carrier, such as sterile water, optionally in admixture with other pharmaceutically acceptable ingredients, such as suspending agents, buffers, pH adjusting agents, preservatives, isotonizing agents.

Sufficient amounts of suitable buffering agents and pH adjusters should be used to adjust the dispersion to neutral to slightly alkaline (up to pH 8.5), preferably in the pH range of 7-7.5. A particular buffer is a salt of a weak acid. Buffers and pH adjusting agents that may be added may be selected from tartaric acid, maleic acid, glycine, sodium lactate/lactic acid, ascorbic acid, sodium citrate/citric acid, sodium acetate/acetic acid, sodium bicarbonate/carbonic acid, sodium succinate/succinic acid, sodium benzoate/benzoic acid, sodium phosphate, tris (hydroxymethyl) aminomethane, sodium bicarbonate/sodium carbonate, sodium hydroxide, benzenesulfonic acid, sodium benzoate/acid, diethanolamine, glucose delta lactone (gluco delta lactone), hydrochloric acid, hydrobromic acid, lysine, methanesulfonic acid, monoethanolamine, sodium hydroxide, tris, gluconic acid, glyceric acid, glutaric acid, glutamic acid, ethylenediaminetetraacetic acid (EDTA), triethanolamine, including mixtures thereof.

Preservatives include antibacterial agents and antioxidants, which may be selected from benzoic acid, benzyl alcohol, Butylated Hydroxyanisole (BHA), Butylated Hydroxytoluene (BHT), chlorobutanol, gallates, hydroxybenzoates, EDTA, phenol, chlorocresol, meta-cresol (metacresol), benzalkonium chloride, tetradecyl-gamma-p-methylpyridinium chloride, mercuric phenylacetate and thimerosal. Free radical scavengers include BHA, BHT, vitamin E and ascorbyl palmitate, and mixtures thereof. The oxygen scavenger comprises sodium ascorbate, sodium sulfite, L-cysteine, acetylcysteine, methionine, thioglycerol, acetone sodium bisulfite, isoascorbic acid, and hydroxypropyl cyclodextrin. Chelating agents include sodium citrate, sodium EDTA and malic acid.

Isotonic or isotonicity agents (isotonicifiers) may be present to ensure isotonicity of the micro-or nanoparticle compositions of the present invention and include sugars such as glucose, dextrose, sucrose, fructose, trehalose, lactose; polyhydric sugar alcohols, preferably trihydric or higher sugar alcohols, such as glycerol, erythritol, arabitol, xylitol, sorbitol and mannitol. Alternatively, sodium chloride, sodium sulfate or other suitable inorganic salts may be used to render the solution isotonic. These isotonic agents may be used alone or in combination. The suspension conveniently comprises 0-10% (w/v), especially 0-6% isotonic agent. Non-ionic isotonicity agents, such as glucose, are important when electrolytes can affect colloidal stability.

The viscosity of the micro-or nanoparticle compositions of polymorph I of TMC278 should be low enough to be administrable by injection, below about 75 mPa-s, or below 60 mPa-s.

The micro-or nanoparticle compositions of the present invention preferably comprise as much polymorph I of TMC278 as is tolerable, such that the injection volume is maintained to a minimum, in particular 3-40% (w/v), or 3-30% (w/v), or 3-20% (w/v), or 10-30% (w/v) polymorph I of TMC 278. In an embodiment, the micro-or nanoparticle composition comprises about 10%, or about 20%, or about 30% (w/v) of polymorph I of TMC 278.

In one embodiment, the aqueous suspension may comprise the following components in weight ratios based on the total volume of the composition:

(a) 3% -50% (w/v), alternatively 10% -40% (w/v), alternatively 10% -30% (w/v) of polymorph I of TMC 278;

(b) 0.5% to 10%, alternatively 0.5% to 2% (w/v) of a humectant;

(c) 0% -10%, alternatively 0% -5%, alternatively 0% -2%, alternatively 0% -1% of one or more buffering agents;

(d) 0% -10%, or 0% -6% (w/v) isotonic agent;

(e) 0% -2% (w/v) of a preservative; and

(f) water for injection q.s. to 100%.

An amount of acid or base may optionally be added to the suspension to bring the pH to a value of about pH 7. Suitable acids or bases may be any physiologically acceptable ones, such as HCl, HBr, sulfuric acid, alkali metal hydroxides, such as NaOH.

The administration of polymorph I of TMC278 of the present invention may be sufficient to treat HIV infection, although in many cases it may be recommended to co-administer other HIV inhibitors, such as other types of HIV inhibitors, in particular those selected from the group consisting of NRTIs, PIs, fusion inhibitors, integrase inhibitors and CCR 5-inhibitors.

In certain instances, treatment of HIV infection may be limited to the administration of micro-or nanoparticle compositions of polymorph I of TMC278, i.e. a monotherapy that is not co-administered with other HIV inhibitors. Such monotherapy may be recommended, for example, when the viral load is relatively low, for example, when the viral load is below about 200 replications per ml, especially below about 100 replications per ml, more especially below 50 replications per ml, especially below the detection limit of the virus. In one embodiment, this type of monotherapy, which may be referred to as supportive therapy, is suitable for use during a certain period of time after initial treatment with an HIV drug combination (especially in combination with any HAART) until the viral load in the plasma reaches the low viral levels mentioned earlier.

In another aspect, the present invention relates to the use of a micro-or nanoparticle composition comprising an anti-virally effective amount of polymorph I of TMC278 for the manufacture of a medicament for use in the maintenance of therapy of a patient infected with HIV, wherein the composition is administered or is to be administered intermittently at a time interval ranging from 1 week to 1 year, or from 1 week to 2 years, or from 1 month to 3 months or any other time interval mentioned herein.

The present invention also provides a method for the long term treatment of a patient infected with HIV, the method comprising:

(a) treating the patient with an HIV inhibitor in combination; followed by

(b) Intermittently administering a micro-or nanoparticle composition comprising an antiviral effective amount of polymorph I of TMC278 as defined herein, wherein the composition is administered at a time interval in the range of 1 week to 1 year, or 1 month to 3 months or any other time interval mentioned herein.

The invention also relates to the use of the micro-or nanoparticle compositions described herein as a medicament for the treatment or prevention of HIV infection, and to the use of the micro-or nanoparticle compositions described herein in the manufacture of a medicament for the prevention or treatment of HIV infection. The present invention also relates to methods of treating a patient infected with HIV comprising administering a therapeutically effective amount of a micro-or nanoparticle composition as described herein.

Polymorph I of TMC278 may be used alone or in combination with other antiviral agents, especially with antiretroviral agents. The present invention therefore also relates to a product comprising (a) polymorph I of TMC278 and b) one or more other antiretroviral compounds, for simultaneous, separate or sequential use as a combined preparation for anti-HIV treatment. Different drugs can be combined with pharmaceutically acceptable carriers to make a single formulation. Accordingly, the present invention also relates to a pharmaceutical composition comprising a pharmaceutically acceptable carrier and (a) a therapeutically effective amount of polymorph I of TMC278 and (b) one or more other antiretroviral agents.

The other antiretroviral compounds include any antiretroviral compound such as suramin, pentamidine, thymopentin, castanospermine, dextran (dextran sulfate), foscarnet (trisodium phosphinocarboxylate); nucleoside Reverse Transcriptase Inhibitors (NRTIs) such As Zidovudine (AZT), didanosine (ddl), zalcitabine (ddC), lamivudine (3TC), stavudine (d4T), emtricitabine (FTC), Abacavir (ABC), amdoxovir (DAPD), elvucitabine (ACH-126,443), AVX 754((-) -dOTC), fozivudine tidylate (FFZT), phosphazide (phosphazide), HDP-990003, KP-1461, MIV-210, racivir (PSI-5004), UC-781 and the like; non-nucleoside reverse transcriptase inhibitors (NNRTIs), such as Delavirdine (DLV), Efavirenz (EFV), Nevirapine (NVP), dapivirine (TMC 120), etravirine (TMC125), DPC-082, (+) -Calcareubin plant extractFetching A, BILR-355; nucleotide reverse transcriptase inhibitors (NtRTIs), such as tenofovir ((R) -PMPA), Tenofovir Disoproxil Fumarate (TDF), and the like; nucleotide competitive reverse transcriptase inhibitors (NcRTIs), such as the compounds of WO 2004/046143; transactivator inhibitors, such as TAT-inhibitors, e.g., RO-5-3335, BI-201, and the like; an REV inhibitor; protease inhibitors such as Ritonavir (RTV), Saquinavir (SQV), lopinavir (ABT-378or LPV), Indinavir (IDV), amprenavir (APV or VX-478), TMC 126, nelfinavir (NFV or AG-1343), atazanavir (BMS 232,632), darunavir (TMCl 14), fosamprenavir (GW433908 or VX-175), breinavir (GW-640385, VX-385), P-1946, PL-337, PL-100, tipranavir (TPV or PNU-140690), AG-1859, AG-1776, Ro-0334649, etc.; invasion inhibitors comprising fusion inhibitors (e.g., Enfuvirtide (T-20)), adhesion inhibitors and co-receptor inhibitors comprising CCR5 antagonists (e.g., Anliviruo, CCR5mAb004, Malaviruo (UK-427,857), PRO-140, TAK-220, TAK-652, Viriviruo (SCH-D, SCH-417,690)) and CXR4 antagonists (e.g., AMD-070, KRH-27315), examples of invasion inhibitors include PRO-542, TNX-355, BMS-488,043, BlockAide/CR^TMFP 21399, hNM01, nonakine, VGV-I; maturation inhibitors are for example PA-457; viral integrase inhibitors, such as raltegravir (MK-0518), elvitegravir (JTK-303 or GS-9137), BMS-538,158; a ribozyme; an immunomodulator; a monoclonal antibody; gene therapy; a vaccine; siRNAs; and antisense RNAs; a microbicide; zinc finger (Zinc-finger) inhibitors.

As used herein, the phrase "substantially" does not exclude "completely," e.g., a composition that is "substantially free of Y may be completely free of Y. The word "substantially" may be omitted from the definition of the invention when necessary. The term "about" in connection with a numerical value is intended to have its ordinary meaning in connection with the numerical value. The word "about" may be substituted with a value of ± 10%, or ± 5%, or ± 2% or ± 1% when necessary. All documents cited herein are incorporated by reference in their entirety.

Examples

Example 1: preparation of polymorph I of TMC278

Polymorph I of TMC278 was prepared by dissolving about 4.2g of TMC278 in 500ml of 2-propanone while stirring and heating the solution to reflux temperature until a clear solution was obtained. After filtering the clear solution, it was washed with ethanol and dry ice (solid CO)₂) Cooling to below 0 deg.C. The solvent was evaporated until a precipitate was obtained. The resulting crystals were dried under vacuum at room temperature overnight.

Example 2: KBr dispersed phase Infrared (IR) Spectroscopy

Polymorph I of TMC278 is mixed with a basic halide and then compressed into a pellet.

The instrument comprises the following steps: nicolet Magna 560^TMFourier Transform IR (FTIR) spectrophotometer

Scanning number: 32

Resolution ratio: 1cm^-1

Wavelength range: 4000-400cm^-1

Baseline calibration: is that

A detector: DTGS with KBr window

Spectroscope: ge on KBr

Basic halide: KBr

Typical absorption bands characteristic of the FTIR spectrum of TMC278 polymorph I are at about 3348, 3274, 2217, 2209, 1477 and 1334cm^-1. Another absorption band was observed at 3190, 1611, 1523, 1509, 1102, 970, 963, 829 and 820cm^-1. See fig. 1.

Typical absorption bands characteristic of the FTIR spectrum of TMC278 polymorph II are in the region of 3316, 2223, 2215, 1483 and 1325cm^-1. Another absorption band was observed at 3201, 1617, 1516,1505. 1303, 1106 and 967cm^-1. See fig. 5.

All the above-mentioned absorption bands have values of. + -. 2cm^-1。

Example 3: powder XRD

X-ray powder diffraction (XRPD) analysis was carried out using a Philips X' PertPRO MPD diffractometer PW3050/60 with a PW3040 generator. The instrument was equipped with a Cu LFF X-ray tube PW 3373/00. Polymorph I of TMC278 was sprayed onto sample holders at zero order background.

The instrument parameters were as follows:

generator voltage: 45kV

Generator amperage: 40mA

Geometry appearance: Bragg-Brentano

The state is as follows: rotating state

The measurement conditions were as follows:

scanning mode: continuous

Scanning range: 3-50 degrees 2 theta

Step length: 0.01675 degree/step

Counting time: 60.59 seconds per step

Self-rotation time: 1 second

Radiation type: CuKa

Radiation wavelength: 1.54056

Incident light path diffraction light path

Procedure, anti-scattering light column: 15mm remote anti-scatter barrier: +

Soller slit: 0.04rad Soller slit: 0.04rad

Beam mask: 15mm Ni Filter: +

Anti-scattering slit: 1 ° detector: x' Celerator

Knife (beam knife): +

TMC278 polymorph I is characterized by: the main diffraction peaks are at 2 θ positions of 9.0 ° ± 0.2 °, 14.3 ° ± 0.2 °, 17.1 ° ± 0.2 ° and 24.2 ° ± 0.2 °. TMC278 polymorph I is further characterized in that: the X-ray powder diffraction peaks were at the 2 θ positions of 11.3 ° ± 0.2 °, 19.1 ° ± 0.2 ° and 27.6 ° ± 0.2 °. See fig. 2.

Polymorph II of TMC278 is characterised by: the main diffraction peaks are at 2 θ positions of 17.6 ° ± 0.2 °, 21.0 ° ± 0.2 °, 25.8 ° ± 0.2 ° and 27.9 ° ± 0.2 °. Polymorph II of TMC278 is further characterized in that: the X-ray powder diffraction peaks are at 2 θ positions of 8.5 ° ± 0.2 °, 12.4 ° ± 0.2 °, 12.9 ° ± 0.2 ° and 24.8 ° ± 0.2 °. See fig. 6.

Peaks in the XRD spectrum may show intensity variations due to various causes, the most important of which is the history of the sample processing procedure.

Example 4: differential Scanning Calorimetry (DSC)

Approximately 3mg of polymorph I of TMC278 was transferred to a standard aluminum TA-instrument sample tray. The sample pan was covered with a suitable lid and the DSC curve was recorded using a TA-instrument Q1000MTDSC equipped with an RCS cooling unit. The following parameters were used:

initial temperature: 25 deg.C

Heating rate: 10 ℃/min

Final temperature: 280 deg.C

Nitrogen flow: 30ml/min

The melting of TMC278 polymorph I occurred at 257.5 deg.C (max peak), the heat of fusion 122J/g. See fig. 3.

The melting of TMC278 polymorph II occurs at 243.2 deg.C (max peak), the heat of fusion is 153J/g, and then polymorph II is polymorphic converted to polymorph I. DSC is not suitable for determining polymorphic compositions of TMC278 due to the occurrence of transformations. See fig. 7.

Example 5: intrinsic dissolution

Using Hanson Research SR6^TMDissolution test apparatus equipped with USP dissolution apparatus 2 as paddle and Polytetrafluoroethylene (PTFE) tablet holder. The concentration of TMC278 in the solution was determined using a UV spectrophotometer. UV analysis in Agilent 8453^TMOn a spectrophotometer using a 2ml cuvette. Single Component Analysis (SCA) was used, and single component assay recorded single wavelength measurements at 266 nm.

Tabletting

Approximately 125mg of the compound was weighed into a mold having a diameter of 0.8 cm. Tablets were formed by compression using a Speca tablet press with 1 ton of pressure. The tablet has a surface area of 0.5026cm². The sheets were adhered to a PTFE tablet holder.

Reference solution for UV analysis

21.8mg of TMC278 were weighed into a 25ml volumetric flask. The product was dissolved in a 1: 1.5 tetrahydrofuran/methanol mixture. A1 ml aliquot was transferred to a 200ml volumetric flask and then diluted to the mark with 0.01N HCl. The concentration of this reference solution was 0.4364 mg/ml.

Dissolution rate

The tablet holder with the tablets therein was placed in a USP2 dissolution bath. The vessel was charged with 500ml of 0.01N HCl/methanol 1: 1 and thermostatically controlled at 37 ℃. The paddle method was used at a speed of 150rpm (revolutions per minute).Samples were taken every 10 minutes and the concentration was determined by UV spectrophotometry. The slope was calculated from the measurement points between 0 and 60 minutes. The slope is expressed as mg%/min. The Intrinsic Dissolution Rate (IDR) was calculated as mg/(cm)²xmin).

Intrinsic dissolution

The average intrinsic dissolution rate of TMC278 polymorph I is 0.3362. The dissolution profile of TMC278 polymorph I is given in figure 4.

The slope was calculated from 0-60 minutes.

The average intrinsic dissolution rate of polymorph II was 0.2886. The dissolution profile of TMC278 polymorph II is given in figure 8.

The slope was calculated from 0-60 minutes.

Solubility in water

The table below shows the solubility of TMC278 polymorph I in buffer pH4, buffer pH2 and 0.01 NHCl.

The solubility of TMC278 polymorph II in buffer pH4, buffer pH2 and 0.01N HCl is given in the table below.

Example 6: preparation of nanosuspensions of polymorph I of TMC278

A250 ml glass flask and ZrO as grinding medium₂The beads were sterilized in an autoclave. A250 ml glass vial was charged with a solution of 5g of drug and 1.25g of Pluronic F108 in 60ml of water for injection. 300g of ZrO with an average particle size of 500 μm were added₂-beads, the bottle being placed on a roller mill. The suspension was micronised at 100rpm over 72 hours. At the end of the milling process, the thick nanosuspension was removed with a syringe and then bottled. The resulting formulation is formulation 1 in the table below. The concentration was determined by HPLC/UV. Diluted to a final concentration of 25mg/ml TMC 278. The resulting suspension was stored protected from light.

Using similar procedures, formulations 2, 3 and 4 were prepared. They were titrated with 1N sodium hydroxide solution to a pH of about 7. In formulations 2, 3 and 4, Lipoid^TMDissolution of EPG in Tween^TM80 (c).

Composition (I)	Preparation 1	Preparation 2	Preparation 3	Preparation 4
					Polymorph I of TMC278	5g	300mg	300mg	300mg
PluronicTMF108	1.25g	-	-	-
					TweenTM 80	-	75mg	75mg	75mg
LipoidTM EPG	-	9.375mg	9.375mg	9.375mg
					Glucose	-	50mg	50mg	50mg
NaH2PO4.1aq	-	-	2mg	2mg
					Citric acid 1aq	-	-	-	1mg
NaOH 1N	-	pH 6.72	pH 6.98	pH 6.99

The following study was performed using male beagle dogs to compare the plasma kinetics of TMC278 after a single Intramuscular (IM) or subcutaneous injection of the nanosuspension at 5mg/kg (formulation 1, see above).

4 healthy male beagle dogs with a body weight range of 8-16kg at the start of the study were used. The dogs were distinguished by numbers of the ear pricks. Intramuscular (IM) administration was performed in the left and right biceps femoris of two dogs. The left and right thoracic regions of both dogs were administered by subcutaneous injection (SC). The injection amount was 2 X0.1ml/kg in all treatment groups. A 20G needle was used.

Blood samples of 3ml were taken from the left jugular vein of all dogs at the following time points: 0h (predose) on day 0, 20min, 1h, 3h, 8h and 24h post-dose, followed by approximately 8 am on days 2, 3, 6, 8, 10, 13, 16, 20, 23, 27, 29, 36, 43, 50, 57, 64, 71, 78, 85 and 92. Blood samples were placed on EDTA (EDTA Vacutte Greiner, Cat. No.454086, Greiner Labortechnik N.V.). Within 2h of taking the blood sample, the blood sample was centrifuged at about 1900Xg for 10 minutes at room temperature to separate the plasma, which was immediately transferred to the second tube and stored in a freezer within 2 hours from the start of centrifugation. TMC278 in each plasma sample was analyzed individually by means of a validated LC-MS/MS method.

Claims (30)

1. A polymorph of (E)4- [ [4- [ [4- (2-cyanoethenyl) -2, 6-dimethylphenyl ] amino ] -2-pyrimidinyl ] amino ] benzonitrile, i.e. TMC278, characterised by the X-ray powder diffraction pattern disclosed in figure 2.
2. 2. A process for the preparation of the polymorph of claim 1, wherein said polymorph is prepared by dissolving TMC278 in Di C_1-4Alkyl ketones and the solution is heated to reflux temperature and then the solution is cooled.
3. 3. A solid pharmaceutical composition comprising as active ingredient the polymorphic form of TMC278 according to claim 1 and a pharmaceutically acceptable carrier.
4. 4. A pharmaceutical composition of micro-or nanoparticles for administration by intramuscular or subcutaneous injection, comprising a therapeutically effective amount of a polymorphic form of TMC278 according to claim 1, comprising a polymorphic form of TMC278 in micro-or nanoparticle form with a surface modifying agent adsorbed to its surface, wherein the surface modifying agent is selected from the group consisting of poloxamers, α -tocopheryl polyethylene glycol succinate, polyoxyethylene sorbitan fatty acid esters and salts of negatively charged phospholipids or acid forms thereof, suspended in a pharmaceutically acceptable aqueous carrier.
5. 5. A micro-or nanoparticle composition as defined in claim 4, wherein the surface modifier is selected from Pluronic^TMF108, vitamin E TGPS, Tweens^TM80 and Lipoid^TMEPG。
6. 6. A micro-or nanoparticle composition as defined in claim 5, wherein the surface modifier is Pluronic^TM F108。
7. 7. A micro-or nanoparticle composition as defined in any of the claims 4-6, wherein the polymorphic micro-or nanoparticles of TMC278 have an average effective particle size in the range of 50nm-500 nm.
8. 8. A micro-or nanoparticle composition as defined in claim 7, wherein the polymorphic micro-or nanoparticles of TMC278 have an average effective particle size in the range of 100nm to 250 nm.
9. 9. A micro-or nanoparticle composition as defined in any of the claims 4-6, wherein the therapeutically effective amount of the polymorphic form of TMC278 is in the range of 0.5-50 mg/day, calculated on the basis of the dose of TMC 278.
10. 10. A micro-or nanoparticle composition as defined in any of the claims 4-6, wherein the therapeutically effective amount of the polymorphic form of TMC278 is in the range of 5 mg/day-50 mg/day, calculated on the basis of the dose of TMC 278.
11. 11. A micro-or nanoparticle composition as defined in claim 4, comprising the following components in weight ratios based on the total volume of the composition:

    (a) 3% -50% (w/v) of a polymorphic form of TMC 278;

    (b) 0.5% -10% of a humectant;

    (c) 0% -10% of one or more buffering agents;

    (d) 0% -10% of isotonic agent;

    (e) 0% -2% (w/v) of a preservative; and

    (f) enough water for injection is added to 100 percent.
12. 12. A micro-or nanoparticle composition as defined in claim 11, comprising the polymorphic form of TMC278 in a weight ratio of 10% to 40% (w/v) based on the total volume of the composition.
13. 13. A micro-or nanoparticle composition as defined in claim 11, comprising a polymorphic form of TMC278 in a weight ratio of 10% to 30% (w/v) based on the total volume of the composition.
14. 14. A micro-or nanoparticle composition as defined in claim 11, comprising from 0.5% to 2% (w/v) of the wetting agent in a weight ratio based on the total volume of the composition.
15. 15. A micro-or nanoparticle composition as defined in claim 11, comprising from 0% to 5% by weight of the one or more buffering agents, based on the total volume of the composition.
16. 16. A micro-or nanoparticle composition as defined in claim 11, comprising from 0% to 2% by weight of the one or more buffering agents, based on the total volume of the composition.
17. 17. A micro-or nanoparticle composition as defined in claim 11, comprising from 0% to 1% by weight of the one or more buffering agents, based on the total volume of the composition.
18. 18. A micro-or nanoparticle composition as defined in claim 11, comprising an isotonicity agent in a weight ratio of 0% to 6% (w/v) based on the total volume of the composition.
19. 19. Use of a micro-or nanoparticle composition as defined in any of claims 4 to 6 or 11 in the manufacture of a medicament for the treatment of HIV infection.
20. 20. Use of a micro-or nanoparticle composition as defined in any of claims 4 to 6 or 11 in the manufacture of a medicament for the prevention of HIV infection.
21. 21. The use of claim 19, wherein the composition is administered, or will be administered intermittently at a time interval ranging from one week to one year, or one week to two years.
22. 22. The use of claim 19, wherein the composition is administered, or is administered intermittently at a time interval ranging from one week to one month, or ranging from one month to three months, or ranging from three months to six months.
23. 23. The use of claim 22, wherein the composition is administered, or is administered intermittently once a month.
24. 24. The use of claim 22, wherein the composition is administered, or is administered intermittently once every three months.
25. 25. Use according to claim 19, wherein the effective amount of the polymorphic form of TMC278 in the composition is selected such that the blood plasma concentration of TMC278 is maintained at a level ranging from 20ng/ml to 90 ng/ml.
26. 26. The use of claim 20, wherein the composition is administered, or will be administered intermittently at a time interval in the range of one week to one year, or one week to two years.
27. 27. The use of claim 20, wherein the composition is administered, or is administered intermittently at a time interval ranging from one week to one month, or ranging from one month to three months, or ranging from three months to six months.
28. 28. The use of claim 27, wherein the composition is administered, or is administered intermittently once a month.
29. 29. The use of claim 27, wherein the composition is administered, or is administered intermittently once every three months.
30. 30. Use according to claim 20, wherein the effective amount of the polymorphic form of TMC278 in the composition is selected such that the blood plasma concentration of TMC278 is maintained at a level ranging from 20ng/ml to 90 ng/ml.