BRPI0903664A2 - Method of obtaining a new crystalline form of lamivudine, its monohydrate hydrochloride salt, pharmaceutical formulations and their uses - Google Patents

Abstract

METHOD OF OBTAINING A NEW CRYSTALLINE FORM OF LAMIVUDINE, ITS MONOHYDRATED CHLORIDRAATE SALT, PHARMACEUTICAL FORMULATIONS AND ITS USES. The present invention is intended for the method of obtaining a solid crystalline form of lamivudine, form VI, its hydrochloride salt monohydrate and its pharmaceutical formulations. In addition, the present application relates to the use of the crystalline forms of lamivudine monohydrate salt in the preparation of drugs indicated as an anti-HIV agent (acquired immunodeficiency syndrome - AIDS).

Description

"METHOD FOR OBTAINING NEW DELAMIVUDINE CRYSTALLINE, ITS MONOIDRATED CHLORIDRATE SALT, PHARMACEUTICAL FORMULATIONS AND THEIR USES"

Field of the Invention

The present invention is directed to the method of obtaining a crystalline solid form of lamivudine, form VI, its hydrochloride hydrochloride salt and pharmaceutical formulations. Additionally, the present application relates to the use of crystalline forms of lamivudine salmonhydrate in the preparation of medicaments indicated as an anti-HIV agent (Acquired Immunodeficiency Syndrome - AIDS).

Background of the Invention

Nucleoside reverse transcriptase inhibitors (NNRTIs) were the first drugs approved for the treatment of AIDS, which were preventing DNA polymerization. Such compounds, structural analogues of intracellular nucleosides modified by replacing a hydroxyl at the 3 'position by a hydrogen atom, are capable of selectively inhibiting the viral enzyme responsible for DNA synthesis from its RNA after host cell infection.

From rational drug planning strategies, other nucleoside analogues have been synthesized to improve biopharmacokinetic properties and potentiate anti-HIV action.

Lamivudine (C8HnO3N3S, CAS No. 134678-17-4) is chemically termed (2'-cis) -4-amino-1- [2- (hydroxymethyl) -1,3-oxothiolan-5-yl] -2 (1H) -pyrimidinone. Alternatively, lamivudin is also systematically recognized by (-) - cis-4-amino-1- (2-hydroxymethyl-1,3-oxothiolan-5-yl) - (1H) -pyrimidin-2-one. Only an enantiomer has proven biological activity, and is known as lamamivudine. With respect to similarity to intracellular nucleosides, lamivudine is a 2'-deoxygenated analog of cytidine where there is also an isosteric substitution of the ribose 3-methylene group for a sulfur atom.

Lamivudine is marketed in a solid formulation (EPIVIR®) by the pharmaceutical company GlaxoSmithKline.

Lamivudine can be shown in two forms (I and II), form I can still have two morphologically distinct crystalline habits: short rods and extremely thin needles. It is characterized by crystallizing in the orthorhombic crystalline system, non-centrosymmetric spatial group P212121, with 20 lamivudine molecules and four water molecules per crystallographic unit cell. As can be assumed, there are five lamivudine molecules for each water, which is the asymmetric crystallographic unit (Robin K. Harris, Race R. Yung, R. Brian Lamont, Robert W. Lancaster, Sean M. Lynn, Susan E. Stanifort). iPolymorphism 'in a novel anti-viral agent: Lamivudine (J. Chem. Soc., Perkin Trans. 2, pp. 2653-2659,1997).

Lamivudine form II crystals are quadrangular combase bipyramids. It is characterized by crystallization in the tetragonal crystalline system, noncentrosymmetric space group P432i2, as a lamivudine molecule in the crystallographic unit cell. As can be characterized by various structural and analytical techniques, including single crystal X-ray diffraction, it is a dehydrated form that has a single molecule constituting the asymmetric crystallographic unit (Robin K. HARRIS, Race R. YEUNG, R. Brian LAMONT, Robert W. LANCASTER, Sean M. LYNN, Susan E. STANIFORTH. "Polymorphism" in a novel anti-viral agent: Lamivudine, J.Chem. Soc., Perkin Trans. 2, pp. 2653-2659, 1997). crystalline form, in which lamivudine forms a salcom saccharate is also known (BANERJEE Rahul; BHATTPrashant M .; RAVINDRA Nittala V .; DESIRAJU Gautam R. Saccharin. Salts of Active Pharmaceutical Ingredients, Their Crystal Structures, and Increased Water Solubilities. Cryst. Growth Des. , vol. 5, no. 6, p.2299-2309, 2005). This form is characterized by crystallization in the orthorhombic crystalline system, non-centrosymmetric spatial group P2i2i2i, with four dimivors of lamivudine and saccharate per crystallographic unit cell. In this structure, lamivudine acts as a cation because the pyrimidine ring's iminic nitrogen is protonated and the saccharinase is the negatively charged molecular species. Thus, the asymmetric crystallographic unit of this crystalline form is composed of one lamivudine and saccharate molecule. This salt is prepared by a co-crystallization process via slow evaporation of a binary solvent mixture consisting of methanol and chloroform (1: 1, vol / vol), where equimolar amounts of lamivudine and saccharin are previously dissolved.

Contrary to the general behavior of most salt-crystallized drugs, lamivudine saccharate is very poorly soluble, and Form II has a more appropriate dissolution profile for use in pharmaceutical formulations than salt (BANERJEE, Rahul; BHATT, Prashant). R. RAVINDRA, Nittala V.; DESIRAJU, Gautam R. Saccharin Salts of Active Pharmaceutical Ingredients, Their Cystic Structures, and Increased Water Solubilities., Cryst. Growth Des., Vol. 5, No. 6, pp. 2299-2309, 2005) Basically, this low solubility is due to the fact that the hydrogen bonds responsible for the intermolecular architecture within the crystalline reticulum prevent the interaction of water molecules with the molecular units of the crystal. A widely used strategy to improve the solubility, and consequently the bioavailability of a as well as providing its manufacturing characteristics, consists of preparing crystalline forms in which the active molecules are negatively charged, usually when the drugs have acid groups prevailing and / or positively, in this case, most of the time, the drugs have a predominant basic character.

One way to prepare a salt from the free molecule involves crystallization of hydrochlorides. Hydrochlorides are highly appreciated for their practicality of preparation, high yield of laboratory synthesis, low percentage of residual impurities, and improved substantial pharmacotechnical properties and solubility compared to nonionized molecular species.

International publication WO 2007/146115 describes that 6-methoxy-8- [4- (1- (5-fluoro) -quinolin-8-yl-piperidin-4-yl) -piperazin-1-yl] -quinoline hydrochloride It is more soluble in water and in biological fluids than the free base, thus having greater bioavailability, as well as promoting the purity of the active pharmaceutical ingredient due to its crystallinity and facilitating the production process of the drug.

Japanese patent application JP 2008044932 reports that 5- {4 - [(6-methoxy-1-methyl-1H-benzimidazol-2-yl) methoxy] benzyl} -thiazolidin-2,4-dione hydrochloride promotes the increase the purity of the drug to 98-99%, and use non-expensive reagents that do not cause environmental impacts.

Nucleophenylcarbetoxycyclohexene derivatives hydrochlorides, substances with analgesic properties, are known to promote the manipulation of the active ingredients which, when in their free form, present difficulties in filtration and deformulation as they crystallize in the form of a gel composed of tangled fines and that, When dried, it becomes electrostatically charged and highly hygroscopic, and in environments with relative humidity greater than 60% a highly viscous mass with the appearance of a consistent syrup is formed.

U.S. Patent 5,041,543 discloses zidovudine, 3'-azido-3'-deoxythymidine (AZT), which was the first antiretroviral drug that effectively provided clinical benefits, with activity against HIV-I and HIV-2, and also first to be approved for the treatment of HIV infections.

US Patent 5,047,407 and European Patent EP382,526, owned by GlaxoSmithKline, claim the preparation of 1,3-oxothiolane-derived compounds and all their geometric and optical isomers, as well as mixtures thereof.

US 5,905,082 describes two polymorphic forms of lamivudine, called form I and form II. Form I is a metastable polymorph which converts, when exposed to a temperature of 179.6 ° C, to form II, which is the stable form of delamivudine. In addition, the US patent also claims a method of preparing lamivudine form I by heating to 45 ° C a suspension of 64.8 mg lamivudine in 200 ml water for the purpose of obtaining a solution which must be cooled to 30 ° C. With this procedure, a mass is deposited and cannot be further agitated when it must be fractionated and cooled to 10 ° C with agitation. Sequentially, 24-hour drying filtration steps at 45 ° C yield the delamivudine Form I crystals. In addition, the US patent discloses how Form II is prepared by heating to reflux a suspension of 10 mg lamivudine in 200 ml denatured alcohol for the purpose of obtaining a clear solution which must be filtered while still hot. Then, about half the volume of the filtrate solvent must be distilled off and then heating is stopped when recognized shape II crystals are seeded into the preconcentrated solution. The already added solution of the form II seed crystals is then cooled from 80 ° C to 25 ° C for one hour, with crystal growth beginning at 79 ° C. Finally, cooling steps, now to 15 ° C , and stirring for one hour, followed by filtration, washing with denatured alcohol and drying, give the lamivudine Form II crystals.

International publication WO 2003/027106 shows that lamivudine form II can also be obtained by suspending 100 g of lamivudine salicylate monohydrate in 600 ml of ethyl acetate, or acetonitrile, stirring for 10 minutes at a temperature range ranging from 30 ° C. To 35 ° C, followed by slow heating to a temperature sufficient to achieve reflux of the used solvent. At the moment when the reflux of the solvent occurs 60 g of triethanolamine in 50 ml of ethyl acetate are slowly added for one hour. Sequentially, the refluxing process is continued for 30 minutes, followed by cooling of the solution and stirring for one hour at 30 ° C, resulting, after filtration, washing and drying in lamivudine Form II crystals.

Furthermore, international publication WO 2007/119248 discloses a new crystalline form of lamivudine, Form III, which may also be used as an active pharmaceutical ingredient in solid formulations. It is characterized by crystallizing into the monoclinic crystalline system, a noncentrosymmetric space group P2i, with two lamivudine molecules and one crystallographic cell-water molecule, where two water molecules are interacting with four lamivudine molecules via hydrogen-class bonds. Thus, there is a lamivudine molecule for each water molecule, which is the asymmetric crystallographic unit of this lamivudine hemidrate. Form III crystals may be prepared in two ways: 1) by dissolving lamivudine in water at 45 ° C, followed by slow cooling of the stirred solution, isolation of the crystalline material from the matrix solution and optionally washing with organic solvent and drying; 2) stirring formI or form II crystals in water at 20-45 ° C, followed by slow cooling of the stirring solution, isolation of the crystalline material from the matrix solution and optionally washing with organic solvent and drying.

International publication WO 2007/119248 claims a form I with lower particle density and flow properties than previously claimed. Thus, proposing a solution to the serious problems regarding the handling of the input during the pharmaceutical formulation. However, it is well known that this form is unstable, constituting an obstacle to the standardization and quality control of finished products and even the raw material containing their form. On the other hand, Form II, which is thermodynamically stable, and Form III have better production and dissolution properties, making the use of these for the composition of solid forms feasible.

As previously mentioned, the hydrochloride preparation is a widely used alternative to circumvent the physicochemical characteristics of drugs that disadvantage drug cross-linking of their crystalline forms containing only the active molecule.

International publication WO 2008/049645 describes a crystalline form of erlotinib, a cytostatic drug used for the treatment of various cancers, is prepared by recrystallization of an aqueous solution of the compound in question, providing purification and ecological benefits as the process of Crystal formation leads to decreased impurities and also due to the fact that organic solvents are not required for the preparation of this crystalline form.

US Patent Application 2005282860 portrays fexofenadine hydrochloride, an antihistamine and bronchodilator drug used for the treatment of allergic episodes and respiratory manifestations, can also be found in a hydrated crystalline form useful as a pharmaceutical ingredient for medicinal products intended for clinical use.

International publication WO 2006/118087 indicates (-) - 2- [4- [2 - [[1S, 2R] -2-hydroxy-2- (4-hydroxyphenyl) -1-methylethyl] amino] ethyl acetate hydrate ] -2,5-dimethylphenoxyethyl, in its polymorphic phase where it is crystallized as a hydrochloride, is attributed excellent stability and solubility, which is stated when these properties are compared with those of other crystalline forms of this drug, which is used to the treatment of urinary incontinence and prostatic hypertrophy, among other pathologies.

Also, the production of solid desibutramine dosage forms, a drug used as an antidepressant agent, anti-obesity tranquilizer due to its demonoamine neuronal reuptake inhibitory effect, is one of the prime examples of pharmacotechnical applications of hydrated hydrate forms, as presented by international publications WO 1994/00047 and WO2004 / 099119, where the pharmaceutically acceptable crystalline form dasibutramine is presented as a hydrated salt, more specifically a sibutramine monohydrate hydrochloride. From all of the foregoing, the Depositor has developed a method of obtaining a stable crystalline form of lamivudine from its salt, featuring pharmacotechnical properties and improved availability.

Description of the Figures

Figure 1 shows the morphologies of lamivudine crystals, images captured on a LEO435 VP scanning electron microscope at 15 kV, with (IA) the bipyramids of Form II and (IB) the stretched plates of lamivudine monohydrate hydrochloride. The bars indicate 100 μm.

Figure 2 shows a molecular view of the crystallographic asymmetric unitity of lamivudine monohydrate hydrochloride at room temperature prepared with ORTEP-3, shows an arbitrary labeling of atoms, which are represented by their respective thermal ellipsoids at a probability of 50%.

Figure 3 shows the X-ray diffractograms of lamivudine hydrochloride hydrochloride, 3A at room temperature, 3B at low temperature, as well as experimental X-ray diffractogram3C at room temperature (intermediate).

Figure 4 illustrates the contents of the room temperature crystallographic unit cell prepared with ORTEP-3.

Figure 5 shows a structural representation prepared with ORTEP-3, showing the supramolecular architecture formed within the crystalline structure of lamivudine monohydrate hydrochloride at room temperature. The double dotted lines illustrate the hydrogen bonds along the direction [100], which directly surround the chloride anion and / or the water molecule, and the hydrogen bond between lamivudine molecules along the direction [102]. The dipole between the protonated imine nitrogen atom of the pyrimidine nucleus N2 and the oxygen atom O2 of the member ring is also graphically represented by two dotted lines. Hydrogen atoms involved in intermolecular interactions were represented as spheres of arbitrary radius. From symmetry operators (represented by lowercase letters that override the labeling of atoms involved in intermolecular contacts) i = -1 + x; y; -1 + z, ii = x; y; -1 + z, iii = 1 + x; y; z, iv = 2 + x; y; - ι + Ζ, ν = 3 - x; 0.5 + y; 1 - z, vi = 1 + x; y; -1 + ζ and vii = 2 - x, 0.5 + y; 1 - z, all units of the structure are constructed from the crystallographic identity, whose atoms do not have any overlapping letters.

Description of the Invention

The present invention is directed to the method of obtaining a crystalline solid form of lamivudine, form VI, its hydrochloride hydrochloride salt and pharmaceutical formulations. Additionally, the present application relates to the use of crystalline forms of lamivudine salmonhydrate in the preparation of medicaments indicated as an anti-HIV agent (Acquired Immunodeficiency Syndrome - AIDS).

In a first embodiment, the present application relates to the method of obtaining lamivudine monohydrate hydrochloride by steps (a) dissolving from about 1 to 40 mg of delamivudine form II in about 1 to 50 ml bidistilled water at temperatures between 35 ° C and 50 ° C. Stirring for 2 to 10 minutes, followed by rest for 10 to 30 minutes or until room temperature is reached, (b) adding about 50 to 500 μΐ of an aqueous hydrochloric acid solution (0.2 to 2% w / v), stirring the solution for 2 to 10 minutes and (c) resting the mixture at room temperature at 25 ° C to 30 ° C until complete evaporation of the solvent matrix, when, then lamivudine hydrochloride hydrochloride crystals are isolated.

In a second embodiment, the present invention provides lamivudine monohydrate hydrochloride with crystallographic X-ray diffraction characterization at room temperature, having the following crystallographic unit cell parameters: a = 5.2927 (2) Å; b = 17,252 (1) Å; c = 6.8518 (4) Å; α = 90.00 °; β = 98.865 (3) ° and γ = 90.00 °. At low temperature, vectors a = 5,2554 (1) Á; b = 17.1944 (6) Å; c = 6.8210 (2) Å; and the angles α = 90.00 °, β = 98.774 (2) ° and γ = 90.00 °.

In addition, the positive charge lamivudine cationic species originated by protonation of the pyrimidine ring nitrogen nitrogen atom, the chloride anionic species and a water molecule make up the crystallographic asymmetric unit.

In addition to the lamivudine cationic species with the proton pyrimidine ring iminitic denitrogen atom, the crystallographic asymmetric unit of lamivudine hydrochloride monohydrate is also composed of the anionic chloride species and a water molecule.

As for crystalline packaging, one-dimensional chains are formed parallel to the crystallographic axis a, containing chloride anions and interspersed water molecules within a structure that can be seen as a framework where lamivudine moieties are anchored.

Specifically, lamivudin monohydrate hydrochloride can be fully recognized by molecular conformation, where the 2 ', 3'-dideoxyribose ring with isosteric substitution of the 3-methylene group by a sulfur atom has a conformationenvelope, where sulfur atom Sl is the displaced from the least square planom formed by atoms C6, 02, C5 and C7, distanced from this plane by 0.806 (5) Â at room temperature and 0.827 (4) At low temperature (the mean square deviation of the adjusted atoms in the least square plane is 0, 0443 ° C at room temperature and 0.0478 ° C at low temperature).

Still with respect to the sulfur atom, it is cis-oriented with respect to the pyrimidine ring, which can be seen as a substituent of the C5 carbon atom within the minimum plane through the four abovementioned atoms.

Already the oxygen atom 03 of the hydroxymethyl arm remains axially oriented, and undoubtedly this atom does not have an eis or trans orientation with respect to the modified 2 ', 3'-dideoxyribose ring intracyclic oxygen atom. Nevertheless, the hydroxyl hydrogen atom H3c, which was clearly located within the electron density distribution map constructed by Fourier differential analysis, is positioned relative to the modified 2 ', 3'-dideoxyribose ring oxygen atom.

In addition, hydrogen atom H3c is located outside the space domain of the C6-C8 bond, which is described by the torsion angle C6-C8-03-H3c (-134 (2) ° at room temperature and -133 (3) ° at low temperature). This displacement of the hydrogen atom H3c away from the middle center of the Csp3-Csp3 bond connecting the hydroxymethyl arm to the modified riboside ring is in the intermolecular hydrogen bond 03-H3c ... 01 which forces the removal of said hydrogen atom in favor of an appropriate geometric orientation between the hydrogen donor (03-H3c) and acceptor (C1 = 01) groups involved in this interaction. This intermolecular contact is responsible for the binding of lamivudine molecules along the direction [102], which are related by translational demetry operations along the crystallographic vectors a and c.

The lamivudine molecule within the lamivudine hydrochloride hydrochloride structure has a hydroxyl group 03-H3cils configured relative to the pyrimidine ring, with a distortion angle 02-C6-C8-03 of 63.7 (4) ° at room temperature and 63 ° C. , 5 (3) ° at low temperature. As for the pyrimidine nucleus, the plane of this heterocyclic ring is almost parallel to the longitudinal axis of the02-C5 bond, as the carbonic group Cl = Ol oriented inversely to the intracyclic oxygen atom of the five-membered ring. Respectively, the torsion angles 02-C5 -N1-C4 (-13.6 (5) ° at room temperature and -13.3 (3) ° at low temperature) and 02-C5-N1-C1 (164.1 (3) ° at room temperature and 163, 8 (2) ° at low temperature) numerically report both characteristics.

Lamivudine monohydrate hydrochloride shows a peculiar arrangement of intermolecular hydrogen bonds within the crystalline reticulum, where chloride anions and water molecules seal along the direction [100] forming a true ribbon to which lamivudine cations are anchored. In fact, chloride anions play a key role in stabilizing crystalline packaging in this long direction because they are hydrogen acceptors in four classic hydrogen bonds, two of the NH ... Cl type and two involving the water molecule (Ow-Hw ... Cl).

The hydrogen bonds involving the N3-H3a ... C11 atoms and the N2 + -H2 ... C11 atoms, both between the lamivudine pyrimidine nucleus and the chloride anion, interact specifically with one amino group hydrogen atom and the atom. positively charged imine nitrogen atom bound to the inorganic species of the heterocyclic ring plane formed by the atoms NI, Cl, OI, N2, C2, N3, C3, and C4, since the atom Cll is distanced from this plane by 0.070 (5 ) At room temperature and by 0.099 (4) Å at low temperature (the mean squared deviation of the eight atoms set in the least square plane is 0.0055 Å at room temperature and 0.0041 Å at low temperature).

In turn, water interconnects related chloride anions by operating translational symmetry along the direction [100], through the hydrogen bonds Ow-Hlw ... Cll and Ow-H2w ... Cll. At low temperature, both hydrogen bonds involving the anionchloride and the water molecule end up being favored geometrically, which can be seen by the narrowing of the distances between the donor atoms and hydrogen acceptors of these interactions (the Ow ... Cll separation for the Ow-Bond bond). Hlw ... Cll measures 3,1170 (4) Â at room temperature and 3,156 (3) Â at temperature of 150 (2) K and for Ow-H2w contact ... Cll measures 3,163 (4) at room temperature and 3,154 (3) Á at a temperature of 150 (2) K) and by increasing the collinearity of the atoms involved, since the Ow-Hlw ... Cll and Ow-H2w ... Cll bond angles expanded from 167 (5) ° and 164 (4) ° at room temperature to 173 (4) ° and 175 (5) ° at a temperature of 150 (2) K, respectively. The decrease in the crystallographic axis value a due to the lowering of temperature is partly due to the intermolecular approximation of water molecules to chloride anions.

One-dimensional chains are formed parallel to the crystallographic axis a, with alternation of chloride anions and water molecules within them, where the protonated lamivudine species are anchored by two hydrogen bonds with the negatively charged inorganic unit. As a consequence, the pyrimidine ring planes are stacked parallel along the direction [100], with an interplanar distance of 3.4 (1) Á at ambient temperature and 3.37 (4) Á at a temperature of 150 (2) K.

An ion-dipole interaction between the modified epentose pyrimidine rings contributes to the stabilization of the stacking of lamivudine molecules in this direction. This contact occurs between the positively charged imine nitrogen atom, N2, which belongs to the pyrimidine ring, and the altered riboside residue endocyclic oxygen atom, 02. Such atoms are separated by 3.038 (4) at room temperature and 3.002 (4) Á at a temperature of 150 (2) K, these values are within the expected optimal range

for the existence of such interaction (approximately 3A). At low temperature, there is a shortening of the distance N2 ... 02, which can be directly linked to the contraction of the crystallographic unit cell parameter a in this thermal condition.

On the other hand, water not only acts as a molecular link between chloride anions within the ribbon growing one-dimensionally along the direction [100]. Unlike hydrogen bonds with chloride anions, in which the water molecule is a hydrogen donor group, it also participates as a hydrogen acceptor in an intermolecular interaction with an adjacent three-dimensional lamivudine molecule, involving the N3-H3b atoms. Ow Hydrogen binding is presented as one of the two contacts responsible for the union of independent chains directed parallel to the crystallographic axis a. Thus, the importance of this alteration for the three-dimensional crystal structuring of lamivudine hydrochloride hydrochloride stands out, together with the connective role of the intermolecular bond 03-H3c ... 01, both interconnecting the direction [102] along the strips formed by repetitive units of [ Cl · (Lam +) - H2O] n.Particularly, this application presents the plans of the pyrimidine rings oriented parallel along the direction [102], due to the hydrogen bonds N3-H3b ... Ow and 03-H3c ... 01, with an orthogonal distance between planes of 1.03 (1) A at room temperature and 1.07 (1) At 150 (2) K, considering that each pyrimidine ring has its distant center7,987 ( 4) From another center in this direction. This latter value corresponds to the determination conducted at room temperature, and the relevant data for structural determination from the reflections collected at 150 (2) K is 7,95 (1) Á.

X-ray powder diffraction analysis of lamivudine hydrochloride hydrochloride of the present invention also demonstrates Bragg bias at the following positions at 2Θ: 10.24; 13.06; 14.04;

16.64; 16.94; 17.70; 19.84; 20.24; 20.44; 22.32; 22.96; 23.62; 24.4625.16; 26.30; 26.82; 27.80; 28.30; 28.68; 29.02; 29.58; 30.62; 31.0632.72; 33.10; 33.64; 33.84; 34.06; 34.26; 34.66; 34.84; 35.20; 35.2835.60; 35.86; 36.36; 37.20; 37.76; 38.26; 38.74; 38.86; 39.11; 39.2839.78; 39.92; 40.16; 40.27; 40.42; 40.76; 41.02; 41.16; 41.33; 41,7841.90; 42.38; 43.02; 43,26; 43.38; 43,82; 44.06; 44.26; 45.32; 45,4846.16; 46.30; 46.46; 46.70; 46.92; 47.14; 47.32; 47.41; 48.14; 48.3448.96; 49.08; 49.36; 49.78 ± 0.2 °.

In a third embodiment, the present invention addresses pharmaceutical formulations containing the new crystalline form VI lamivudine monohydrate hydrochloride, such dosage forms may be presented in solid forms such as powders, granules, capsules, tablets, tablets, coated tablets and optionally tablets. associated with pharmaceutically acceptable carriers.

In a fourth embodiment, the patent application is for use of the novel crystalline forms of delamivudine monohydrate hydrochloride in the preparation of drugs indicated as an anti-HIV agent (acquired immunodeficiency syndrome - AIDS).

Examples

Preparation of Lamivudine Monohydrate Hydrochloride

10 mg lamivudine form II was dissolved in 5 ml of 40 ° C bidistilled water with stirring for 5 minutes. The solution was then left to stand for 10 minutes or until room temperature was reached at around 25 ° C. Finally, a 250 μΐ aliquot of a standard deconcentrated aqueous hydrochloric acid solution (1% w / v) was added, stirring the solution for a period of 5 minutes. The resulting mixture was kept at room temperature at 25Â ° C until complete evaporation of the solvent matrix, when lamivudine hydrochloride monohydrate crystals were isolated.

Essay

Monocrystalline X-ray Diffraction Analysis

From the morphological point of view, only a single crystalline type of lamivudine monohydrate hydrochloride was found after thorough inspection by polarized light photon microscopy and scanning electron microscopy. Crystals showing stretched plate aspects have been reported.

A well-formed, translucent, nonrefringing plaque-like crystal of lamivudine monohydrate hydrochloride, dimensions 0.584 χ 0.098 χ 0.027 mm, was selected for the single-crystal X-ray diffraction experiment. Data collection was performed at room temperature (298 (1) K) and at low temperature (150 (2) K) with the aid of an Enraf-Nonius Kappa-CCD diffractometer (95 mm CCD camera as detector and angular geometry κ). using a sealed Mo tube (MoKa = 0.71073 Á) generated at 60 kV e33 mA and graphite monochromed. An OxfordCryosystem Nitrogen blower was used for lowering and maintaining a temperature of 150 (2) K.

A total of 7886 Bragg reflections were collected at room temperature, with a three redundancy, in an angular range at θ from 3.83 ° to 23.80 ° (Miller index variation: from -6 to 5 to h, from -19 to19 to k from -7 to 7 to (i). In the case of the low-temperature experiment, 8661 Bragg reflections were collected at room temperature, also with a three redundancy, in a 2Θ angular range of 6.04 ° to 54.90 ° (Miller index range: -5 to 6 parah from -21 to 20 to k, from -8 to 8 to Ζ). The completeness in Gmax was 99.3% and 95.5% at 298 (1) K and 150 (2) K, respectively, and the crystallographic unit cell parameters presented were obtained over the entire set of collected reflections, the same. used to resolve and refine the structure of delamivudine monohydrate hydrochloride. The COLLECT program (Enraf-Nonius, COLLECT. NoniusBV, Delft, The Netherlands, 1997-2000) was used as a computational interface tool for the collection of reflections, and the HKL Denzo-Scalepack (OTWINOWSKI and MINOR, HKL Denzoand Scalepack. : Methods in Enzymology, vol 276, edited by CWCarter Jr. and RM Sweet, New York: Academic Press, pp. 307-326, 1997) was used for data integration and scaling.

The Gaussian absorption correction method was implemented during structural refinements (COPPENS et al., Calculation of absorption corrections for camera and diffractometerdata. Acta Crystallogr., Vol. 18, p. 1035-1038, 1965), provided that the hydrochloride monohydrate structure of Lamivudine is composed of sulfur and chlorine atoms. The minimum and maximum values of the transmission factors (Tmin and Tmax) of 0.881 and 0.986, respectively, were perceived at room temperature. At low temperature, the calculated values of Tmin and Tmax were 0.941 and 0.987, respectively.

The structure of lamivudine monohydrate hydrochloride was solved by the direct methods of solving the diffraction-radiation phase problem, implemented in the SHELXS-97 computer program (SHELDRICK, SHELXS-97. Program for Ciystal StructureResolution. University of Gottingen: Gottingen, Germany, 1997).

All non-hydrogenoid atoms were located in the electron density distribution map within the crystallographic unit cell, obtained by Fourier synthesis after statistical recovery of the diffracted wave phases. This initially obtained model was refined by the complete matrix least squares method, minimizing the difference between the observed and calculated structure factors in F2, using, for this purpose, the SHELXL-97 computer program (SHELDRICK, SHELXL-97. Program for Crystall Structures Analysis. University of Gottingen: Gottingen, Germany, 1997). Over 1862 independent reflections (Rint = 0.1337) at room temperature and over 2547 independent reflections (Rint = 0.1170) at 150 (2) K, 166 parameters were refined in one of the structural determinations, depending on their thermal condition.

All hydrogen atoms covalently bonded to carbon atoms were located at stereochemical positions with fixed distances of 0.93 Å (aromatic C-H bond), 0.97 Å (C-H bond within methylene groups) and 0.98 Â (C-link within metino groups). Both amine hydrogen atoms were given the same treatments as hydrogen atoms bonded to carbon atoms, with fixed distances of 0.86 Å for bonding — H. Already the imine proton, lamivudine hydroxyl hydrogen, and the two hydrogen atoms in the molecule of water were located on the electronic density distribution map constructed by Fourier differential analysis, and their positional parameters were refined.

For all hydrogen atoms, an isotropic thermal displacement model was adopted, and each hydrogen atom bonded to carbon and nitrogen atom was assigned a fixed thermal parameter equal to the equivalent isotropic thermal parameter to which it was attached, plus 20% of this value. In the case of hydrogen atoms attached to oxygen atom, the same treatment above isotropic thermal parameters was conducted, however, it was considered an increase of 50% of the equivalent isotropic thermal parameter of the atom to which the hydrogen atom was attached.

For the structure determined at room temperature, the maximum and minimum unmodified electron density values within the crystallographic unit cell were 0.179 and -0.187 e.A3, respectively, and the final merit parameters were: S = 1.029, R1 [I> 2σ (7)) = 0.0412, wR2 (I> 2a (I)) = 0.0937, R1 (for all data) = 0.0573 and wR2 (for all data) = 0.1022.

In the case of the structure determined at 150 (2) K, unmodified maximum and minimum electron density values within the crystallographic unit cell equal to 0,432 and -0,520 e.A3, respectively, S = 1,065, R1 (7> 2σ (2 )) = 0.0493, wR2 (I> 2a (I)) = 0.1113, R1 (for all data) = 0.0522 and it> R2 (for all data) = 0.1141. The computer program ORTEP-3, version 2.0 (FARRUGIA, ORTEP-3 for Windows - a version of ORTEP-III with a Graphical User Interface (GUI). J. Appl Crystallogr., Vol. 30, p. 565, 1997) was used to make the structural graphical representations, all illustrations made with the structure determined at room temperature. As atoms with atomic number greater than that of the canopy, sulfur and chlorine, configured in a non-centrosymmetric structure, the refinement of the Flack parameter was possible (FLACK, On enantiomorph-polarity estimation (Acta Crystilogr., Sect. A, vol. 39, pp. 876-881, 1983).

A value of -0.10 (10) was refined based on 892 Friedel pairs obtained at room temperature, and a value of 0.11 (8) was achieved over 1177 Friedel pairs collected at low temperature. Absolute configuration was performed, where carbons C5 and C6, considering the labels of the atoms given in Figure 2, presented the configurations S and R, respectively.

Powder X-ray Diffraction Analysis

Crystals of lamivudine monohydrate hydrochloride were reduced in size and were later fitted to a grooved glass slide for exposure to the X-ray beam. X-ray powder diffraction experimentation was performed on a Rigaku Denki powder diffractometer with Θ-2Θ geometry, with scintillation detector, 50 kV and 100 mA generator, radiation from a Cu rotary anode (CuKa, y = 1 , 54187 À) and continuous scan mode. The scan was conducted at room temperature (298 (1) K), in the range of 8 to 50 ° at 2 °, with a pitch of 0.02 ° and a scanning speed of 1,000 ° / min.

The experimental X-ray diffractogram was the same as the theoretical X-ray diffraction pattern simulated within the MERCURY program (BRUNO et al., New software for searching the Cambridge Structural Database and visualizing crystal structures. Acta Crystallogr. Sect. B, vol. 58 , pp. 389-397, 2002) from the crystal structure of lamivudine hydrochloride hydrochloride determined by demonocrystalline X-ray diffraction at room temperature.

No predicted X-ray diffraction signals were detected (2Θ) not predicted for delamivudine monohydrate hydrochloride, which could come from other crystalline phases, for example, typical reflections of lamivudine Form I, Form II and Form III, although preferential orientation effect of the crystals was introduced during the preparation of the crystalline sample of this new crystalline form of lamivudine intended for the powder X-ray diffraction experiment. Such artifact of the technique had already been expected because the crystals of delamivudine monohydrate hydrochloride present characteristic morphology, such as stretched plates. Thus, it can be stated that the sample analyzed for lamivudine monohydrate hydrochloride is of high purity and that the method of preparation of the novel crystalline form of this anti-HIV nucleoside reverse transcriptase inhibitor drug is highly effective and unendurable.

The results presented in this invention provide for the preparation of a new crystalline form of lamivudine, a lamivudine hydrochloride hydrate, which was precisely characterized by single crystal and powder X-ray diffraction analysis. Few laboratory steps that do not require excessive time, do not require organic solvents and do not require sophisticated instrumentation lead to the efficient preparation of this crystalline form of lamivudine, thus qualifying the synthesis method as fast, simple, economical, robust and environmentally viable.

Table 1 summarizes the structural information regarding the determination of the crystallographic unit cell of lamivudine hydrochloride hydrochloride.

<table> table see original document page 24 </column> </row> <table>

The crystalline phase transition evidently does not occur between temperatures of 298 Ke 150 K (Table 1). However, the crystallographic unit cell parameters a, b and c and the crystalline monoclonic form of lamivudine monohydrate hydrochloride are shortened at low temperature, as is the angle β also at 150K (Δατ «ηρ. = 298k - aisok = 0.037 A; AlbTemp . = fc> 298k - fel50k = 0.057 Å; ACTemp = C298k "ClSOk = 0.030 A; Δβτ« ηρ. = PlSOk-p298k = 0.091 °).

The reduction of certain intermolecular distances within the crystalline reticulum of lamivudin monohydrate hydrochloride as a consequence of the lowering of the temperature from 298 K to 150 K explains this decrease in the values of the crystallographic unit cell descriptors at low temperature. Table 2 shows the fractional coordinates x, y and ζ (x10 "4) of the nonhydrogenous atoms of delamivudine monohydrate hydrochloride and their equivalent isotropic parameters (Ueq) of thermal displacement (À2 χ IO 3) of the nonhydrogenous atoms of lamivudinated hydrochloride monohydrate determined at temperatures of 298 (1) K and 150 (2) K.

Table 2

<table> table see original document page 25 </column> </row> <table>

Table 3 shows the fractional coordinates x, y and z (χ 10ˉ4) of the hydrogen atoms of delamivudine monohydrate hydrochloride and the respective isotropic parameters (Ueq) of thermal displacement (Á2 χ 10 ˉ3) of hydrogen atoms of the crystallographic identity of lamivariate hydrochloride monohydrate. at temperatures of 298 (1) K and 150 (2) K.

Table 3

<table> table see original document page 26 </column> </row> <table>

Table 4 presents the geometric details of the intermolecular hydrogen bonds within the lamivudine monohydrate hydrochloride crystal structure. The capital letters DeA refer to the hydrogen donor and acceptor, respectively, and the length values and bonding angles are of both structural determinations at temperatures of 298 (1) K and 150 (2) K, with the Symmetry Code: -1 + x, y, z; b Symmetry code: 1 + x, y, 1+ z; c Symmetry code: x, y, 1 + z; d Symmetry code: -1 + x, y, 1 + z.Table 4

<table> table see original document page 27 </column> </row> <table>

Claims (30)

1. METHOD OF OBTAINING A SOLIDACRISTALINE FORM OF LAMIVUDINE, characterized in that it comprises the steps (a) dissolving lamivudine form II in about 1 to 50 ml bidistilled water at temperatures between 35 ° C and 50 ° C, under agitation. for 2 to 10 minutes, followed by standing for 10 to 30 minutes or until room temperature is reached, (b) adding an aqueous hydrochloric acid solution, stirring the solution for 2 to 10 minutes, and (c) mixing the mixture. temperature at 25 ° C to 30 ° C until complete evaporation of the solvent matrix. Method of obtaining a solid-crystalline form of lamivudine according to claim 1, characterized in that lamivudine form II comprises amounts of from about 1 to 40 mg. METHOD FOR OBTAINING A LAMIVUDINE SOLID-CRYSTALLINE FORM according to claim 1, characterized in that the aqueous hydrochloric acid solution comprises amounts of about 50 to 500 μΐ with standard concentration. Method of obtaining a solid form of LAMIVUDINE LAMIVUDINE according to claim 3, characterized in that the standard concentration of the aqueous hydrochloric acid solution comprises about 0.2 to 2% w / v. 5. LAMIVUDINE SALT, characterized in that it comprises form VI, monohydrate hydrochloride. 6. LAMIVUDINE MONOIDRATED CHLORIDRATE according to Claim 5, characterized in that the lamivudine monohydrate hydrochloride form comprises the crystallographic characterization by X-ray diffraction at room temperature, having the following crystallographic unit cell parameters: a = 5,2927 (2) Á; b = 17,252 (1) Å; c = 6.8518 (4) Å; α = 90.00 °; β = 98.865 (3) ° and γ = 90.00 °. LAMIVUDINE MONOIDRATED CHLORIDRATE, according to claim 5, characterized in that the lamivudine monohydrate hydrochloride form comprises the crystallographic characterization by X-ray diffraction at low temperature, having the following crystallographic unit cell parameters a = 5,2554 ( 1) À; b = 17.1944 (6) Å; c = 6.8210 (2) Å; and the angles α = -10 90.00 °; β = 98.774 (2) ° and γ = 90.00 °. LAMIVUDINE MONOIDRATED CHLORIDRATE according to claims 6 and 7, characterized in that the crystallographic unit cell parameters still comprise ± 0.1 Å and / or ± 0.1 ° standard deviations. LAMIVUDINE MONOIDRATED CHLORIDRATE according to claims 5 to 8, characterized in that the crystallographic asymmetric unit of lamivudin monohydrate hydrochloride comprises an anionic chloride species and a water molecule. LAMIVUDINE MONOIDRATED CHLORIDRATE according to claims 5 to 9, characterized in that the crystalline packaging comprises one-dimensional chains formed parallel to the crystallographic axis a, containing anionschloride and water molecules interspersed within the structure. LAMIVUDINE MONOIDRATED CHLORIDRATE according to claims 5 to 10, characterized in that the molecular conformation comprises the 2 ', 3'-dideoxyribose ring with isosteric substitution of the 3'-methylene group by a sulfur atom. LAMIVUDINE MONOIDRATED CHLORIDRATE according to claim 11, characterized in that the molecular conformation further comprises an envelope conformation, wherein the sulfur atom Sl is the most displaced of the least square plane formed by the atoms C6, 02, C5 and C7, distanced from this plane by 0.806 (5) Â at room temperature and by 0.827 (4) Â at low temperature. LAMIVUDINE MONOIDRATED CHLORIDRATE according to claims 5 to 12, characterized in that the sulfur atom is understood to be oriented with respect to the pyrimidine ring. LAMIVUDINE MONOIDRATED CHLORIDRATE according to claims 5 to 13, characterized in that the oxygen atom 03 of the hydroxymethyl arm is axially oriented. LAMIVUDINE MONOIDRATED CHLORIDRATE according to Claims 5 to 14, characterized in that the hydroxyl hydrogen atom H3c is comprised of the modified 2 ', 3'-dideoxyribose ring oxygen atom. LAMIVUDINE MONOIDRATED CHLORIDRATE according to claim 15, characterized in that the hydrogen atom H3c is still facing outward of the C6-C8 bond spatial domain by the torsion angle C6-C8-03-H3c (--134 ( 2) ° at room temperature and -133 (3) ° at low temperature). LAMIVUDINE MONOIDRATED CHLORIDRATE according to claims 5 to 16, characterized in that the lamivudine molecule within the lamivudine monohydrate hydrochloride structure comprises a hydroxyl group 03-H3c and is configured with respect to the pyrimidine ring with a torsion angle-02- C6 -C8-03 of 63.7 (4) ° at room temperature and 63.5 (3) ° at low temperature. LAMIVUDINE MONOIDRATED CHLORIDRATE, according to claims 5 to 17, characterized in that the pyrimidinic nucleus is partially parallel to the axis-long axis of the 02-C5 bond and oriented inversely to the intracyclic deoxygen atom of the five-membered ring. LAMIVUDINE MONOIDRATED CHLORIDRATE according to claim 18, characterized in that the pyrimidinic nucleus still comprises torsion angles 02-C5-N1-C4 (-13.6 (5) ° at room temperature and -13.3 (3 ) ° at low temperature) and 02-C5-N1-C1 (164.1 (3) ° at room temperature and 163.8 (2) ° at low temperature). LAMIVUDINE MONOIDRATED CHLORIDRATE according to claims 5 to 19, characterized in that lamivudine monohydrate hydrochloride comprises an arrangement of intermolecular hydrogen bonds within the crystalline reticulum, where chloride ions and water molecules alternate along the direction [100]. ] forming a tape to which lamivudine cations seancor. LAMIVUDINE MONOIDRATED CHLORIDRATE, according to claims 5 to 20, characterized in that lamivudine monohydrate hydrochloride comprises water molecules that interconnect chloride anions by symmetriatranslational operation along the direction [100] through the Ow-HlwCll hydrogen bonds and Ow-H2wC11. LAMIVUDINE MONOIDRATED CHLORIDRATE according to claims 5 to 21, characterized in that the direction [100] comprises an ion-dipole interaction between the pyrimidine ring and the modified pentose. LAMIVUDINE MONOIDRATED CHLORIDRATE, according to claim 22, characterized in that the interaction is comprised between the positively charged imine nitrogen atom, N2, and the altered residuoriboside endocyclic oxygen atom, 02. LAMIVUDINE MONOIDRATED CHLORIDRATE, according to claim 23, characterized in that the denitrogen and oxygen atoms comprise separations of 3.038 (4) A at room temperature and 3.002 (4) Á at a temperature of 150 (2) K. LAMIVUDINE MONOIDRATED CHLORIDRATE, according to claims 5 to 24, characterized in that the molecule still comprises a hydrogen acceptor activity in the intermolecular interaction with an adjacent three-dimensional lamivudine molecule involving the N3-H3b ... Ow. LAMIVUDINE MONOIDRATED CHLORIDRATE according to claims 5 to 25, characterized in that the planes of the pyrimidine rings oriented parallel to the direction [102] comprise orthogonal distances between the planes of 1.03 (1) at ambient temperature and 1 , 07 (1) At the temperature of 150 (2) K. LAMIVUDINE MONOIDRATED CHLORIDRATE according to claims 5 to 26, characterized in that the powder X-ray diffraction comprises Bragg reflections at 2Θ positions: -10.24; 13.06; 14.04; 16.64; 16.94; 17.70; 19.84; 20.24; 20.44; 22.32, -22.96; 23.62; 24.46; 25.16; 26.30; 26.82; 27.80; 28.30; 28.68; 29.02, -29.58; 30.62; 31.06; 32.72; 33.10; 33.64; 33.84; 34.06; 34.26; 34.66, -34.84; 35.20; 35.28; 35.60; 35.86; 36.36; 37.20; 37.76; 38.26; 38.74, -38.86; 39.11; 39.28; 39.78; 39.92; 40.16; 40.27; 40.42; 40.76; 41.02, -41.16; 41.33; 41.78; 41.90; 42.38; 43.02; 43,26; 43.38; 43,82; 44.06, -44.26; 45.32; 45.48; 46.16; 46.30; 46.46; 46.70; 46.92; 47.14; 47.32, -47.41; 48.14; 48.34; 48.96; 49.08; 49.36; 49.78 ± 0.2 °. 28. PHARMACEUTICAL FORMULATIONS according to claims 5 to 27, characterized in that the formulations comprise solid forms. 29. PHARMACEUTICAL FORMULATIONS according to claim 28, characterized in that the solid forms comprise powders, granules, capsules, tablets, tablets, coated tablets and / or dragees, optionally associated with pharmaceutically acceptable ovules. The uses of the crystalline forms of chlamydro-hydrochloride LAMIVUDINE according to claims 1 to 29, characterized in that it comprises the preparation of drugs indicated as an anti-HIV agent (Acquired Immunodeficiency Syndrome - AIDS).