Polymorphic and other crystalline forms of cis-FTC

DESCRIPTION

Polymorphic forms and other crystalline forms of cis-FTC

FIELD OF THE INVENTION 5

The present invention relates to polymorphic forms of (-) - cis-FTC (4-amino-5-fluoro-1- (2-hydroxymethyl) -1,3-oxathiolan-5-yl) -2 (1H) -pyrimidinone ).

BACKGROUND OF THE INVENTION 10

The success of several synthetic nucleosides such as AZT, D4T, DDI and DDC in inhibiting HIV replication in vivo or in vitro led researchers, at the end of the 1980s, to design and analyze nucleosides that replace the carbon atom in the 3 'position of the nucleoside with a heteroatom. Norbeck et al. revealed that (±) -1- [cis (2,4) -2- (hydroxymethyl) -4-dioxolanyl] thymine (designated as (±) -15 dioxalane-T) has modest activity against HIV (EC50 of 20 μM in ATH8 cells) and is not toxic to non-infected control cells at a concentration of 200 μM, Tetrahedron Letters 30 (46), 6246, (1989). European Patent Application Publication No. 337,713 and US Pat. No. 5,041,449, transferred to BioChem Pharma, Inc., reveal 1,3-dioxolanes-substituted in 2-substituted, 4, racemic, which exhibit antiviral activity. PCT applications published under PCT numbers US91 / 09124 and PCT 20 US93 / 08044 reveal isolated β-D-1,3-dioxolanyl nucleosides for the treatment of HIV infection. WO 94/09793 discloses the use of β-D-1,3-dioxolanyl nucleosides for the treatment of HIV infection.

U.S. Patent No. 5,047,407 and European Patent Application Publication No. 0 382 526, 25 also transferred to BioChem Pharma Inc., reveal that different racemic 1,3-oxathiolane-substituted 2-substituted nucleosides have antiviral activity and specifically describe that the racemic mixture of 2-hydroxymethyl-5 - (cytosin-1-yl) -1,3-oxathiolane (hereinafter referred to as BCH-189) has approximately the same activity against HIV as AZT, with a lower toxicity. The (-) enantiomer of BCH-189 (U.S. Patent No. 5,539,116, issued to Liotta et al.), Known as 3TC, is currently sold commercially in the US. for the treatment of HIV in humans. See also document EP 513.200 B1.

Also, it has been revealed that (-) - (cis) -FTC (4-amino-5-fluoro-1- (2- (hydroxymethyl) -1,3-oxathiolan-5-yl) -2- (1H) -pyrimidinone (2R-cis) or β-L-2-hydroxy-methyl-5- (5-fluorocytosin-1-yl) -1,3-oxathiolane) has a potent activity against HIV; see Schinazi et al., "Selective Inhibition of Human Immunodeficiency viruses by Racemates and Enantiomers of cis-5-Fluoro-1- [2- (Hidroxymethil) -1,3-Oxathiolane-5-yl] Cytosine", Antimicrobial Agents and Chemotherapy , November 1992, pages 2423-2431. See also US Pat. No. 5,814,639, 5,914,331, 5,210,085 and 5,204,466; and WO 91/11186 and WO 92/14743. The chemical structure of (-) cis-FTC is shown below: 40

<figref>image 1</figref>

C8H10FN3O3S

Molecular Weight: 247.25

Due to the commercial importance of 1,3-oxathiolane nucleosides such as FTC, various procedures for their production in patents and in the scientific literature have been described. The substituents on chiral carbons (the specified purine base or pyrimidine (designated as substituent C5) and CH2OH (referred to as substituent C2)) of the 1,3-oxathiolan nucleosides may be cis (on the same side) or trans ( on opposite sides) with respect to the oxathiolane ring system). Both racemates, cis and trans, 45 consist of a pair of optical isomers. Therefore, each compound has four individual optical isomers. The four optical isomers are represented by the following configurations (orienting the rest of oxathiolane in a horizontal plane so that the rest -S-CH2- is behind): (1) cis (also designated as β), with both groups "in the upper part ”, which is the naturally occurring L-cis configuration; (2) cis, with both groups "at the bottom", which is the β-cis configuration that does not occur naturally; (3) trans (also designated α configuration) with the substituent C2 "on the part

upper ”and the substituent C5“ in the lower part ”; and (4) trans, with the C2 substituent "on the bottom" and the C5 substituent "on the top". The two cis enantiomers together are designated as a racemic mixture of β enantiomers, and the two trans enantiomers together are designated as a racemic mixture of α enantiomers. In general, separation of the cis racemic optical isomer pair from the trans racemic optical isomer pair is quite conventional. Separation or otherwise obtaining individual enantiomers of the cis configuration is significantly more difficult. For 3TC and FTC, the desired stereochemical configuration is the β-L isomer.

The numbering scheme of the 1,3-oxathiolane ring in FTC is shown below:

<figref>image2</figref>

Routes to condense the 1,3-oxathiolane ring with a protected base

U.S. Pat. No. 5,204,466 discloses a process for condensing a 1,3-oxathiolane with a protected pyrimidine base using tin chloride as Lewis acid, which provides a virtually complete β stereomer selectivity. See also Choi et al., "In Situ Complexation Directs the Stereochemistry of N-Glicosylation in the synthesis of Oxathiolanyl and Dioxolanyl Nucleoside Analogues", J. Am. Chem. Soc. 1991, 213, 9377-9379. The use of tin chloride creates residues and undesirable by-products during the reaction, which are difficult to remove. fifteen

Several US patents reveal a process for the preparation of 1,3-oxathiolane nucleosides by condensation of a 1,3-oxathiolane intermediate, which has a chiral ester in the 2-position of the ring, with a protected base in the presence of an acid of Lewis based on silicon. Subsequently, the ester in position 2 must be reduced to the corresponding hydroxymethyl group to generate the final product. See US Pat. No. 5,663,320, 5,864,164, 5,693,787, 5,696,254, 5,744,596 and 5,756,706.

U.S. Pat. No. 5,763,606 discloses a process for predominantly producing 1,3-oxathiolane cis-2-carboxylic or thiocarboxylic-nucleosides which includes coupling a desired silylated purine or pyrimidine base 25 with a bicyclic intermediate in presence of a Lewis acid. U.S. Pat. No. 5,272,151 describes a process for the preparation of 1,3-dioxolane nucleosides that includes reacting a 2-O-protected-5-0-acylated-1,3-dioxolane with a purine or pyrimidine base with an oxygen or a protected nitrogen, in the presence of a titanium catalyst. 30

Choi et al., "In Situ Complexation Directs the Stereochemistry of N-Glicosylation in the synthesis of Oxathiolanyl and Dioxolanyl Nucleoside Analogues", J. Am. Chem. Soc. 1991, 213, 9377-9379, describe that no coupling of the 1,3-oxathiolane, with the pyrimidine base protected, with HgCl2, Et2AlCl or TiCl2 (O-isopropyl) 2 (see footnote 2). Choi et al. They also describe that the reaction between 35 anomeric 1,3-oxathiolane acetates with silylated cytosine and virtually any common Lewis acid, other than tin chloride, results in the formation of inseparable mixtures of N-glycosylated anomers.

U.S. Pat. No. 5,922,867 discloses a process for the preparation of a dioxalan nucleoside 40 that includes glycosylation of a purine or pyrimidine base with an oxymethyl-4-halo-1,3-dioxalane-protected in 2.

U.S. Pat. No. 5,914,331, 5,700,937, 5,827,727 and 5,892,025, among others, issued to Liotta et al., Describe the coupling of 1,3-oxathiolanes, disclosed in this description, with 5-45 silylated fluorocytosine , in the presence of SnCl4, to form the β (-) isomer of FTC and, optionally, the removal of protective groups.

Routes to provide the 1,3-oxathiolane nucleoside in the desired stereo configuration

50

In U.S. Pat. No. 5,914,331, 5,700,937, 5,827,727 and 5,892,025, among others, issued to Liotta et al., Describe specific procedures for preparing FTC with the desired stereo configuration, in a substantially pure form. In one embodiment, the C5'-hydroxyl group of a mixture of nucleoside racemates is reacted with an acyl compound to form C5 'esters, in

which the nucleoside is at the "carbinol" end of the ester. The desired enantiomer can be isolated by treating the racemic mixture with an enzyme that hydrolyzes the desired enantiomer (followed by extraction of the polar hydrolyzate with a polar solvent) or by treatment with an enzyme that hydrolyzes the unwanted enantiomer (followed by removal of the unwanted enantiomer with a polar solvent). Enzymes that catalyze the hydrolysis of 1,3-oxathiolane pyrimidine 5 nucleosides include pig liver esterase, porcine pancreatic lipase, Amano PS-800 lipase, subtilisin and α-chymotrypsin.

Cytidine deoxycytidine deaminase can be used to solve racemic mixtures of 2-hydroxymethyl-5- (cytosin-1-yl) -1,3-oxathiolane and its derivatives, which include 2-hydroxymethyl-5- (5-fluoro-cytosine -1-il) -1,3-10 oxathiolane. The enzyme catalyzes the deamination of the rest of cytosine to uridine. One of the enantiomers of 1,3-oxathiolane nucleosides is a preferred substrate for cytidine deoxycytidine deaminase. The enantiomer that is not converted to uridine (and, therefore, is still basic) is extracted from the solution with an acid solution. Citidine deoxycytidine deaminase can be isolated from rat liver or from human liver, or it can be expressed from recombinant sequences in a prokaryotic system such as E. Coli.

Chiral chromatography can also be used to resolve cis-FTC enantiomers. For example, US Pat. No. 5,892,025, issued to Liotta et al., Discloses a procedure for resolving a combination of cis-FTC enantiomers by passing cis-FTC through a chiral column of acetylated β-20 cyclodextrin.

Polymorphic Characterization

The ability of a compound to exist in different crystalline structures is known as polymorphism. These distinct crystalline forms are known as "polymorphic modifications" or "polymorphs." Although polymorphs have the same chemical composition, they differ in packaging and geometric arrangement and have different physical properties, such as melting point, shape, color, density, hardness, deformability, stability, dissolution, and the like. Depending on their temperature-stability relationship, two polymorphs could be monotropic or enantiotropic. For a monotropic system 30, the relative stability between the two solid phases remains unchanged as the temperature changes. In contrast to this, in an enantiotropic system there is a transition temperature at which the stability of the two phases is reversed. (Theory and Origin of Polymorphism, in "Polymorphism in Pharmaceutical Solids", (1999) ISBN: 8247-0237).

35

Several compounds that have polymorphism have been described. As an early example, Gordon et al., In US Pat. No. 4,476,248, revealed and claimed a new crystalline form of the drug ibuprofen, as well as a process for its production. It was described that the new crystalline form improved the manufacturing capacity of ibuprofen. It has also been described that a structure more closely related to FTC, 3TC ((-) - cis-4-amino-1- (2-hydroxymethyl-1,3-oxathiolan-5-yl) - (1H) -40 pyrimidin -2-one; lamivudine) exists in more than one crystalline form. Jozwiakowski, MJ, Nguyen, NT, Sisco, JM, Spancake, CW "Solubility Behavior of Lamivudine Crystal Forms in Recrystallization Solvents", J. Pharm. Sci., 85, 2, pages 193-199 (1996). See also US Pat. No. 5,905,082, issued to Roberts et al., On May 18, 1999, under the title "Crystalline Oxathiolane Derivatives", and its PCT analogue, PCT / EP92 / 01213, which describes two polymorphic forms of 3TC. Roberts et al., Reveal that a polymorph is obtained when 3TC crystallizes in an aqueous solution. A second polymorph is obtained when 3TC crystallizes in a non-aqueous medium, or when the first form melts and is allowed to cool, or when the first form is crushed or ground. Both polymorphic forms have absorption bands, melting temperatures and unique crystalline energies.

50

The (-) - cis-FTC produced by the procedures described above, has a distinct crystalline form, designated herein as Form I of (-) - cis-FTC. The angular positions (two theta) of the characteristic peaks in a powder X-ray diffraction diagram of Form I of (-) cis FTC, shown in Figure 7, are: 14.1º ± 0.1º, 19, 9º ± 0.1º, 20.2º ± 0.1º, 20.6 ± 0.1º, 21.0º ± 0.1º, 22.4º ± 0.1º, 28.5º ± 0.1º, 29.5º ± 0.1º and 32.6º ± 0.1º. 55

Additional polymorphs and other crystalline forms of FTC could have commercial value in their manufacture or in other applications. Therefore, an objective of this invention is to provide new polymorphic forms and other crystalline forms of FTC.

60

Another objective is to provide new procedures for the preparation and isolation of polymorphic forms and other crystalline forms of FTC.

Still, another object of the invention is to provide therapeutic uses of FTC polymorphs and other FTC phases. 65

SUMMARY OF THE INVENTION

The present invention is directed to a polymorphic form of (-) - cis-FTC characterized by a solid state transition at 96-112 ° C.

5

The present invention is set forth in the attached independent claim. The sub-embodiments of the present invention are set forth in the attached dependent claims.

The solid phases of (-) - cis-FTC, which are hereby designated as (-) - amorphous FTC and Forms II and III of (-) - cis-FTC) are intended to be distinguished from Form I (-) - cis-FTC by X-ray powder diffraction patterns, thermal properties and manufacturing methods. A hydrated crystalline form of (±) -cis-FTC (ie, racemic cis-FTC), and a dehydrated form of the hydrate are also provided, and can be similarly distinguished from other forms of cis-FTC by diffraction patterns X-ray powder, thermal properties and manufacturing methods. These forms of FTC can be used in the manufacture of other forms of FTC, or in pharmaceutical compositions. Particularly preferred uses of these forms are in the treatment of HIV or hepatitis B.

Form II of (-) - cis-FTC can be obtained by fusing Form I of (-) - cis-FTC and allowing the fusion to recrystallize at a temperature close to the melting point of Form I. Form III of ( -) - cis-FTC can be obtained by cooling Form II of (-) - cis-FTC below the thermodynamic transition temperature 20 of Forms II and III. The (-) - amorphous cis-FTC can be obtained by rapid liquid cooling of (-) - cis-FTC. The hydrated crystalline form of (±) -cis-FTC is a sesquihydrate, and could be obtained by dissolving (±) -cis-FTC in water and recrystallizing FTC. The dehydrated form of sesquihydrate can be obtained by removing the hydration water from sesquihydrate.

25

BRIEF DESCRIPTION OF THE FIGURES

Figure 1 is a typical DSC thermogram (differential scanning calorimetry) of Form I of (-) - cis-FTC, with an endotherm at 151 ° C, obtained by heating at a rate of 10 ° C / min.

Figure 2 is a DSC thermogram of Form I of cis (-) FTC obtained by heating at 1 ° C / min. 30

Figure 3 is a DSC thermogram of Form I of cis (-) FTC obtained by heating at 2 ° C / min.

Figure 4 is a DSC thermogram of Form I of cis (-) FTC obtained by heating at 5 ° C / min.

Figure 5 is a DSC thermogram of Forms II and III of (-) - cis-FTC.

Figure 6 is a DSC thermogram showing that the transition from Form II to III is reversible. 35

Figure 7 is a diagram of PXRD (X-ray powder diffraction) of Form I of (-) - cis-FTC.

Figure 8 is a PXRD (X-ray powder diffraction) diagram of Form II of (-) - cis-FTC.

Figure 9 is a PXRD diagram of Form III of (-) - cis-FTC.

Figure 10 is a PXRD diagram of (±) -cis-FTC sesquihydrate.

Figure 11 is a PXRD diagram of a dehydrated form of racemic cis-FTC 40 sesquihydrate.

Figure 12 is a DSC thermogram of (±) -cis-FTC sesquihydrate and (-) - cis-FTC.

Figure 13 is a scan by TGA (thermogravimetric analysis) of (±) -cis-FTC sesquihydrate.

Figure 14 is a relationship between free energy and temperature for three polymorphs of (-) - cis-FTC (Forms I, II and III). Stable phases are represented by a continuous line and metastable phases 45 with a dotted line.

Figure 15 is a DSC thermogram of (-) - amorphous cis-FTC obtained by melting and freezing a crystalline sample of (-) - cis-FTC. The final heating rate was 10 ° C / min.

DETAILED DESCRIPTION OF THE INVENTION 50

Two new polymorphic forms of (-) - cis-FTC, the amorphous phase of (-) - cis-FTC, a new crystalline form of hydrated (±) -cis-FTC and a hydrate form of (±) - are provided dehydrated cis-FTC, in pharmaceutical compositions. The different forms can be distinguished from other phases of (-) - cis-FTC and (±) -cis-FTC by X-ray diffraction diagrams, thermal properties and the procedures by which they have been generated. These forms of FTC, together with the amorphous phase, can be used as intermediates in the manufacture of FTC, or they can be formulated in pharmaceutical compositions, as in the present invention, and used for the treatment of HIV or hepatitis B.

The two polymorphic forms of (-) - cis-FTC used in this invention are designated Forms II and III of 60 (-) - cis-FTC and are characterized by powder X-ray diffraction diagrams, in Figures 8 and 9. These forms would contrast with Form I of (-) - cis-FTC, which is the polymorphic form of (-) - cis-FTC prepared by the procedures described in the background section of this document. Form I of (-) - cis-FTC can be characterized by the powder X-ray diffraction diagram, shown in Figure 7, or by the peaks at the diffraction angles given in the background section of this document. 65

The hydrated form of (±) -cis-FTC is characterized by powder X-ray diffraction diagrams, in Figure 10. This form would contrast with the (±) -cis-FTC manufactured in the prior art. The thermal properties of the different forms are summarized in the following table:

<dl><dt> Type of transition T Phase transition </dt><dd> Approximate temperature (ºC) Temperature range (ºC) </dd></dl>

<dl><dt> TIm </dt><dd> 151 148-153 </dd></dl>

<dl><dt> TIIm </dt><dd> 166 162-167 </dd></dl>

<dl><dt> TIIIm </dt><dd> Not observed in this investigation Not observed </dd></dl>

<dl><dt> Tamorfog </dt><dd> 67 heating rate +/- 3ºC at 10ºC / min </dd></dl>

<dl><dt> IT, IIt </dt><dd> 130 (calculated) N / A </dd></dl>

<dl><dt> TII, IIIt </dt><dd> 102 96-112 </dd></dl>

<dl><dt> Thidratod </dt><dd> &gt;30 - </dd></dl>

<dl><dt> Tracematom </dt><dd> 190 185-192 </dd></dl>

Thus, the disclosure provides: 5

a) two polymorphs and an amorphous phase of (-) - cis-FTC, as characterized by dust X-ray diffraction analysis, physical properties and manufacturing methods;

b) a hydrated crystalline form of (±) -cis-FTC, and the dehydrated form of the (±) -cis-FTC hydrate, as characterized by dust x-ray diffraction analysis, physical properties and manufacturing methods ;

c) processes to make the phases of (-) - cis-FTC and the crystalline forms of (±) -cis-FTC;

d) therapeutic and / or pharmaceutical compositions of the phases of (-) - cis-FTC and the crystalline forms of (±) -cis-FTC, optionally in the presence of a pharmaceutically acceptable carrier; and

e) new therapeutic uses for the phases of (-) - cis-FTC and the crystalline forms of (±) -cis-FTC, especially in the treatment of viral diseases such as HIV or hepatitis B.

Form II of (-) - cis-FTC

Form II of (-) - cis-FTC is observed when Form (-) - cis-FTC is melted and allowed to recrystallize. 20 Like all polymorphs, Form II can be characterized by the powder diffraction diagram it presents when it is subjected to powder X-ray crystallography. The angular positions (two theta) of the characteristic peaks of the powder X-ray diffraction diagram of Form II of (-) - cis-FTC, shown in Figure 8, are: 14.7º ± 0.1º, 16 , 7º ± 0.1º, 19.6º ± 0.1º, 21.1º ± 0.1º, 21.8º ± 0.1º, 24.6º ± 0.1º and 25.6º ± 0.1º. 25

Form II of (-) - cis-FTC can also be characterized by its melting temperature and / or heat of fusion: Form II of (-) - cis-FTC has a melting temperature of approximately 166 ° C under pressure Atmospheric and normally has a heat of fusion in the range of approximately 15-19 kJ / mol. It is known that the heat of fusion may vary depending on the experimental conditions. 30

Alternatively, Form II of (-) - cis-FTC can be characterized by its enantiotropic behavior and by the procedure by which it is manufactured. Form II of (-) - cis-FTC is enantiotropic with the polymorphs of Form I and Form III of (-) - cis-FTC, in the sense that there is a transition temperature below and above which reverses the order of stability. Due to this enantiotropic behavior, Form II of (-) - cis-FTC could be prepared from Form I of (-) - cis-FTC or Form III of (-) - cis-FTC. In the examples presented in this document, Form II of (-) - cis-FTC was obtained:

(1) by heating (-) - cis-FTC (Form I) above its melting temperature (approximately 151 ° C 40 for Form I) and keeping it at this high temperature. After cooling slowly, the molten (-) - cis-FTC recrystallized to Form II, and took the crystalline form of Form 11, at temperatures above the thermodynamic transition temperature between Forms II and III;

(2) by heating Form III of (-) - cis-FTC above the thermodynamic transition temperature of Forms II and III, which ranges between approximately 96 ° C and approximately 112 ° C (because Form II is enantiotropic with the Form III of (-) - cis-FTC).

Thus, Form II of (-) - cis-FTC can be obtained when Form I of (-) - cis-FTC is melted and the temperature of the molten material is kept below the melting temperature of Form II but above the transition thermodynamic temperature between Forms II and III. Notably, a similar transition from Form II is not observed when Form 11 is heated above its melting point

(approximately 166 ° C) and allowed to cool slowly. Somehow, Form II simply recrystallizes Form II. However, Form II would not recrystallize from the same molten material if it ceased to cool; instead, it would be an amorphous phase.

Therefore in one embodiment the disclosure provides Form II of (-) - cis-FTC, preferably in substantially pure form, as characterized by any of the above methods. In another embodiment, the disclosure provides Form II of (-) - cis-FTC substantially in the absence of Form I of (-) - cis-FTC. In yet another embodiment, the disclosure provides Form II of (-) - cis-FTC substantially in the absence of Form III of (-) - cis-FTC. The disclosure also provides a pharmaceutical composition comprising Form II of (-) - cis-FTC, which further comprises a pharmaceutically acceptable carrier.

Form III of (-) - cis-FTC

Because Form II of (-) - cis-FTC undergoes a solid state transition to Form III of (-) - cis-15 FTC, this form is obtained from Form D of (-) - cis -FTC when the temperature of Form II of (-) - cis-FTC falls below the transition temperature, which ranges from about 96 ° C to about 112 ° C. Form III of (-) - cis-FTC is another polymorph of (-) - cis-FTC and can be characterized by the powder diffraction diagram that it presents when it is subjected to powder X-ray crystallography. The angular positions (two theta) of the characteristic peaks in the powder X-ray diffraction diagram of Form III of (-) - cis-FTC, shown in Figure 9, are as follows: 14.5º ± 0 , 1st, 16.7º ± 0.1º, 19.6º ± 0.1º, 20.4º ± 0.1º, 21.4º ± 0.1º, 21.7º ± 0.1º, 25.2º ± 0.1º and 26.2º ± 0.1º.

Form III of (-) - cis-FTC can also be characterized by the procedures for its manufacture. Due to the enantiotropic behavior of Form III with Form II, Form III of (-) - cis-25 FTC can be prepared from Form II of (-) - cis-FTC, cooling Form II of (- ) -cis-FTC below the solid state transition temperature of Forms II and III, and thus causing a solid state transition from Form II of (-) - cis-FTC. It is assumed that Form III of (-) - cis-FTC can also be prepared directly from Form I of (-) - cis-FTC, with Form II as intermediate, melting Form I and cooling slowly the molten material at a temperature below 30 solid state transition temperature for Forms II and III. Due to this stability below its solid state transition temperature, Form III of (-) - cis-FTC can also be characterized by the temperature range in which it presents a solid state transition, but is preferably characterized , at the lower end of this range (that is, approximately 96 ° C at atmospheric pressure). 35

Therefore in one embodiment the disclosure provides Form III of (-) - cis-FTC, preferably in substantially pure form. In another embodiment, the disclosure provides Form III of (-) - cis-FTC substantially in the absence of Form I of (-) - cis-FTC. In another embodiment, the disclosure provides Form III of (-) - cis-FTC substantially in the absence of Form II of (-) - cis-FTC. The disclosure also provides a pharmaceutical composition comprising Form III of (-) - cis-FTC, which further comprises a pharmaceutically acceptable carrier.

(±) -cis-FTC sesquihydrate

45

(±) -cis-FTC sesquihydrate is a crystalline form of racemic cis-FTC that is obtained when it dissolves (±) -cis-FTC in water and recrystallizes. Notably, the hydrate is only produced from cis-FTC racemate and is not produced from pure (-) - cis-FTC. The (±) -cis-FTC sesquihydrate can be characterized by the powder diffraction diagram it presents when it is subjected to powder X-ray crystallography. The angular positions (two theta) of the characteristic peaks in the 50 x-ray powder diffraction diagram of the (±) -cis-FTC sesquihydrate, shown in Figure 10, are: 11.5º ± 0.1º, 13 , 4º ± 0.1º, 19.1º ± 0.1º, 20.3º ± 0.1º, 20.8º ± 9.1º, 21.5º ± 0.1º, 21.9º ± 0.1º and 30.9º ± 0.1º.

TGA analyzes confirm a sesquihydrate of (±) -cis-FTC. The sesquihydrate begins to lose its hydration water by evaporation at approximately 30 ° C at atmospheric pressure. 55

The (±) -cis-FTC sesquihydrate can also be characterized by one of the procedures for its preparation. Preferably, the (±) -cis-FTC sesquihydrate is prepared simply by dissolving (±) -cis-FTC in water and recrystallizing the dissolved FTC into a hydrated crystalline form. Heat may be used during dissolution to increase the amount of FTC that dissolves. The (±) -cis-FTC may be present in a pure racemic mixture of cis-FTC, or as an impurity of a composition that mostly comprises (±) -cis-FTC or (-) - cis-FTC. When present as an impurity, the (±) -cis-FTC preferably comprises at least about 4% by weight of the composition of (+) - cis-FTC or (-) - cis-FTC (this it is, if present as an impurity of (-) - cis-FTC, the FTC preferably comprises at least 2% by weight of the enantiomer (+) and, if present as an impurity of (+) - cis- FTC, FTC 65

preferably comprises at least 2% by weight of the enantiomer (-)).

A thermograna of DSC of (±) -cis-FTC is shown in Figure 12. Recrystallized FTC is a sesquihydrate, as analyzed by DSC, TGA and PXRD.

5

Therefore, the present disclosure provides (±) -cis-FTC sesquihydrate, preferably in substantially pure form. The disclosure also provides (±) -cis-FTC sesquihydrate substantially in the absence of Form I of (-) - cis-FTC, or substantially in the absence of other hydrated and dehydrated crystalline forms of cis-FTC, (-) - cis -FTC or (+) - racemic cis-FTC. Also provided herein is a pharmaceutical composition comprising (±) -10 cis-FTC sesquihydrate, further comprising a pharmaceutically acceptable carrier.

After evaporation of the (±) -cis-FTC sesquihydrate hydration water, a dehydrated crystalline form of racemic cis-FTC is generated. The racemic cis-FTC obtained in this way can be characterized by the powder diffraction diagram it presents when it is subjected to 15 x-ray powder crystallography. The angular positions (two theta) of the characteristic peaks of the powder X-ray diffraction diagram of the dehydrated racemic cis-FTC, shown in Figure 11, are: 12.3º ± 0.1º, 14.0º ± 0.1º , 20.7º ± 0.1º, 22.6º ± 0.1º, 23.3º ± 0.1º and 25.5º ± 0.1º. Dehydrated racemic cis-FTC has a melting temperature of approximately 190 ° C at atmospheric pressure, and a heat of fusion of approximately 23 kJ / mol. twenty

The DSC thermogram of Figure 12 corresponds to the (±) -cis-FTC sesquihydrate. As confirmed by TGA analysis, the large endotherm, at approximately 80 ° C, was due to the fact that (±) -cis FTC sesquihydrate was losing its hydration water. The second endotherm, at 190 ° C, corresponds to the melting of the dehydrated racemic cis-FTC. 25

Therefore the disclosure provides (±) -cis-FTC dehydrated. The disclosure also provides the (±) -cis-FTC dehydrated substantially in the absence of Form I (-) - cis-FTC or substantially in the absence of other hydrated and dehydrated crystalline forms of cis-FTC, (-) - cis-FTC or (+) - racemic cis-FTC. A pharmaceutical composition is also provided comprising (±) -cis-FTC dehydrated, which further comprises a pharmaceutically acceptable carrier.

(-) - amorphous cis-FTC

An amorphous form of (-) - cis-FTC is obtained when the molten (-) - cis-FTC rapidly cools to a temperature below about 40-50 ° C, thereby avoiding any transition to Forms II or III of (-) - cis-FTC. A thermogram of amorphous (-) - cis-FTC DSC is shown in Figure 15, which shows that the glass transition temperature of this phase is 67 ° C.

Therefore, the disclosure also provides an amorphous (-) - cis-FTC, preferably in substantially pure form. The disclosure also provides (-) - amorphous cis-FTC substantially in the absence of forms I, II and / or III of (-) - cis-FTC. A pharmaceutical composition is also provided comprising (-) - amorphous cis-FTC, which further comprises a pharmaceutically acceptable carrier.

Definitions 45

As used herein, the term "substantially pure", when used in relation to a phase or crystalline form of FTC, refers to a phase or crystalline form of FTC having a purity greater than about 95%. This means that the polymorphic or hydrated form of FTC does not contain more than about 5% of any other compound and, in one embodiment, does not contain more than 50 of about 5% of any other phase or crystalline form of FTC (either racemic, (-), (±), cis or trans). In other embodiments, the term "substantially pure" refers to a phase or crystalline form of FTC having a purity greater than about 96%. Still, in another embodiment, the term "substantially pure" refers to a phase or crystalline form of FTC having a purity greater than about 97% or 99%. 55

Similarly, the term "substantially in the absence of a second component", when used in relation to a phase or crystalline form of FTC, refers to a phase or crystalline form of FTC that does not contain more than about 5% of the second component More preferably, the term "substantially in the absence of a second component" refers to a phase or crystalline form of FTC 60 that does not contain more than about 4% of the second component and, even more preferably, no more than about 3 % or 1% of the second component.

The characteristic positions of the peaks in the powder X-ray diffraction diagrams are presented, for the crystalline forms, in terms of angular positions (two theta), within a 65

permissible variability of plus or minus 0.1º. The "US Pharmacopeia, p. 1843-1844 (1955) ”specifies this permissible variability. It is intended that the variability of plus or minus 0.1 ° be used when comparing two powder X-ray diffraction diagrams. In practice, if a peak of the diffraction diagram of a diagram is assigned a range of angular positions (two theta), which is a position of the measured peak, plus or minus 0.1 °, and a peak of the diffraction diagram from another diagram you are assigned a range of 5 angular positions (two theta), which is the position of the measured peak, plus or minus 0.1º, and if those intervals of the peak positions overlap, the two peaks are considered to have the same angular position (two theta). For example, if it is determined that a peak of the diffraction diagram of a diagram has a peak position of 5.20º, for comparative purposes, the permissible variability allows the assignment of the peak to a position in the range of 5.10º-5 , 30th. If it is determined that a comparison peak of another 10 diffraction diagram has a peak position of 5.35º, for comparative purposes, the permissible variability allows the peak to be assigned a position in the range of 5.25º-5.45º. Because there is an overlap between the two ranges of peak positions, the two peaks, which are being compared, are considered to have the same angular position (two theta).

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It will be understood that, throughout this specification, the word "comprise" or variations such as "comprises" or "comprising" implies the inclusion of a particular, complete or stage element, or of a group of specific, complete or stages, but not the exclusion of any other determined, complete or stage element, or of any other group of determined, complete elements or stages.

20

EXAMPLES

Materials and methods

The starting materials of (-) - cis-FTC for all analyzes, unless otherwise specified, were obtained by combining and treating two batches of (-) - cis-FTC as follows. A flask was charged with 1109 g of (-) - cis-FTC and 2750 ml of ethyl acetate. This suspension was stirred at room temperature for two hours, filtered and washed with 550 ml of ethyl acetate. The filter cake was dried in a vacuum oven, overnight, at 50 ° C and at a pressure of approximately 0.27 KPa (2 mm Hg). All solvents were of HPLC quality and were used under reception conditions. An HPLC assay 30 of the starting (-) - cis-FTC indicated a purity of 98.8%.

Differential scanning calorimetry (DSC): DSC experiments were performed using one of the following instruments:

35

DSC studies were performed using a TA Instruments DSC 2920 device (with cooling system). Samples of approximately 5 mg were placed in sealed aluminum trays. The DSC cell was purged with 30 ml / min of nitrogen. The heating rate was 10 ° C / min, unless otherwise indicated. The temperature and heat flow calibrations were performed with an indium pattern under the same experimental conditions. 40

DSC measurements were made on a Mettler DCS30 device (Mettler Instrument, Highstown, NJ) equipped with a data analyzer (STARe, Mettler Instrument). The samples (approximately 2-5 mg) were sealed in standard 40 μl aluminum trays, with a single hole in the lid. An empty sample type tray was used as a reference. The samples were scanned at 1-10 ° C / min, with a 50 ml / min purge of dry nitrogen. The DSC was calibrated for heat flow and temperature.

Thermogravimetric analysis (TGA): TGA studies were performed with a TA Instruments TGA 2950 apparatus. Approximately 5 mg samples were placed in open platinum trays and the sample was exposed at a heating rate of 10 ° C / min. fifty

Variable temperature PXRD: The diffractometer (XDS 2000, Scintag, Sunnyvale, CA) consisted of a 4 kW generator (45 kV voltage and 40 mA current) with a copper anode tube, a liquid nitrogen cooled Ge detector (GLP-10195/07-S, EG&amp;G ORTEC, Oak Ridge, TN), a data analyzer (MicroVax 3100, Digital Equipment Corporation, Ontario, Canada), a thermal stage (Scintag) and temperature controller (Microstar, Research Inc., Minneapolis, MN). The samples were placed in a sample holder forming a thin layer and the scanning was carried out at a speed of 1 per minute without rotation.

Thermal stage microscopy (HSM): Polarized light microscopy was performed using an Olympus BX60 microscope equipped with a Mettler-Toledo FP82HT thermal stage. A thin layer of sample 60 was placed on a slide and heated at 10 ° C / min. Thermal events were captured with ImagePro® software.

Recrystallization procedure: Approximately 5 g of (-) - cis-FTC was placed in a round bottom flask and heated in the temperature range of 155 ° C to 160 ° C, for 30 min, with 65

agitation. The sample was cooled in the flask at room temperature, under ambient conditions.

Solubility in equilibrium: The equilibrium solubility values ​​were obtained using an excess of solid in a capped flask that was stirred in a water bath with temperature control, at 25 ° C, for 52 h. The residual solid material was identified, after having equilibrated the sample, by microscopy 5 in thermal plate and PXRD. The supernatant material was filtered through 0.45 µm membrane filters before dilution for HPLC analysis.

Grinding: (-) - cis-FTC was milled in a Fitzpatrick mill at high speed (4000 rpm), with hammer advance, with a 000 band sieve. The drug was passed once through the filter and collected in A 10 plastic bag.

Hydrate Formation: A supersaturated solution (0.5 g / ml) of Form I was prepared at 50 ° C. This solution was subsequently cooled to room temperature, with stirring, for approximately 2 h. The precipitated solid was filtered under vacuum and air dried. This solid was analyzed by HPLC, DSC, PXRD and TGA. These analyzes revealed that the solid was a sesquihydrate of (±) -cis-TFC.

Crystallization: (-) - cis-FTC was dissolved in one of the following solvents: methanol, ethyl acetate, water, tetrahydrofuran and acetone. Each suspension was boiled for approximately 15 minutes and immediately filtered through a 0.45 µm nylon filter. The supernatant was stirred at room temperature until it crystallized. At the point of crystallization, the suspensions were filtered and the filter cakes were collected. The filter cake was placed on a glass disk, covered with a paper towel without threads and placed in a hood under ambient conditions, for 2 days.

Example 1: DSC characterization of Forms I and II of (-) - cis-FTC 25

The thermal events of the Form I polymorph of (-) - cis-FTC were observed at heating rates of 10, 1, 2, and 5 ° C / min. These thermograms are shown in Figs. 1, 2, 3 and 4, respectively. Sample sizes were 6.8400 mg, 5,290 mg, 5,0430 mg or 5,1500 mg, respectively. 30

The endotherm at 151 ° C corresponds to the melting temperature of Form I of (-) - cis-FTC. This endotherm was presented at all heating rates studied. The heat of fusion associated with the melting of this phase is 25 kJ / mol. This fusion is followed by recrystallization from a solid with a higher melting temperature, Form II. The presence of the high temperature endotherm (162 ° C) 35 depended on the heating rate. Specifically, as the heating rate decreased, the probability of the endotherm at high temperature increased. Also, the value of the heat of fusion increased for the high temperature endotherm, as the heating rate decreased. These observations are consistent with the fact that, at slower heating rates, the liquid recrystallizes to a greater extent. This heating rate dependent endotherm indicated that Form I could undergo recrystallization after melting at 151 ° C and that the resulting crystalline form melts at approximately 162 ° C. The phase that melts at 162 ° C was designated "Form II of (-) - cis-FTC".

Example 2: DSC characterization of Forms II and III of (-) - cis-FTC 45

The thermal events of Form I of (-) - cis-FTC, after their fusion and subsequent cooling to room temperature, were observed by DSC. A sample of 5,5500 mg of Form I of (-) - cis-FTC was heated at 160 ° C, a temperature that is just above the melting temperature of Form I, and subsequently cooled again to 25 ° C. . When reheated in the DSC device using a heating rate of 10 ° C / min, the endotherm at 151 ° C of Form I of (-) - cis-FTC was not observed. However, endotherms appeared at 102 ° C and 162 ° C, as shown in Figure 5. The endotherm at 102 ° C corresponded to a solid state transition from Form III of (-) - cis-FTC to Form II of (-) - cis-FTC, as shown in Figure 6. PXRD data (Figures 7 and 9), collected above and below the transition at 102 ° C, confirmed the interpretation of the DSC. The endotherm at 162 ° C corresponded to the fusion of Form II of (-) - cis-FTC. It was confirmed by HPLC that there was no change in the potency associated with these thermal events.

Example 3: DSC characterization of (-) - amorphous cis-FTC

60

A sample of 7.315 mg of Form I of FTC was heated at 180 ° C, at a rate of 5 ° C / min, in the DSC device. Subsequently, the sample was cooled to -20 ° C, at a rate of -20 ° C / min. This sample, when reheated at the rate of 10 ° C / min, showed a displacement in the baseline associated with the glass transition at approximately 67 ° C. This displacement in the baseline occurred, both during the heating cycle and the cooling cycle, which confirmed that it was due to the 65

glass transition Figure 15 contains a DSC thermogram of (-) - amorphous cis-FTC.

Example 4: Observations with HSM of forms I, II and III of (-) - cis-FTC

The assignment of Forms I, II and III based on the thermal events observed during the analysis by DSC was consistent with the observations with HSM. Under the microscope, the material of Form I appeared as plates, at room temperature. After heating at 160 ° C, at a rate of 10 ° C / min, Form I melted into a clear liquid. After cooling this liquid, acicular needles crystallized from the molten material, which had a darker appearance than Form I. Upon reheating, these needles underwent a change in birefringence, which began at approximately 102 ° C and 10 ended at approximately 115 ° C. Eventually, the needles melted at 166 ° C.

Example 5: Effect of grinding on the crystalline form

Two batches of Form I of (-) - ground cis-FTC were prepared: one by manual crushing in a mortar with mace for 5 minutes and another by milling in a Fitzpatrick mill. Although not measured quantitatively, the optical microscope revealed that the particle size of the crushed (-) - cis-FTC appeared as the smallest, followed by that obtained using the Fitzpatrick mill and subsequently by the (-) - cis-FTC not ground. The DSC thermograms of the sample milled with the Fitzpatrick mill and of the (-) - cis-FTC not ground, had only one endotherm at 151 ° C. The crushed (-) - cis-FTC had two endotherms, at 151 ° C and 162 ° C. 20 The PXRD diagram of the (-) - cis-FTC crushed at room temperature was the same as the Form I diagram, indicating that there was a conversion of Form I to Form II during the DSC experiment. These data together indicate that grinding, according to the conditions described, does not affect the crystalline form of (-) - cis-FTC when starting from Form I.

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Example 6: Effect of heat on the crystalline form

The PXRD diagrams for Form III at 25 ° C and 95 ° C are shown in Figure 9. However, a PXRD diagram of this sample, acquired at 120 ° C, was different from the diagram acquired at 95 ° C. The change in the PXRD diagram over this temperature range was consistent with the endotherm obtained at approximately 102 ° C by analyzing the DSC thermogram (Figure 6), and confirms that the endotherm at 102 ° C was caused by a transition in solid state or a change of crystalline structure.

The PXRD diagram, measured at 120 ° C was the same as that obtained at 160 ° C. However, after cooling the sample again to 25 ° C, the PXRD diagram was the same as in Form III of (-) - cis-FTC. The crystalline form that exists above 102 ° C and melts at 162 ° C was identified as Form II. The PXRD diagram of the Form I material did not change to the melting temperature of 151 ° C.

Example 7: Analysis of thermodynamic stability 40

The fusion data of Forms I, II and III of (-) - cis-FTC are summarized in Table I. Based on these data, the thermodynamic relationship between Forms I and II was established. These forms are enantiotropically related and the calculated transition temperature is 130 ° C.

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Table I: Transition temperature data for four crystalline forms of (-) - cis-FTC

<dl><dt> Shape </dt><dd> Transition temperature (ºC) Heat of fusion (kJ / mol) </dd></dl>

<dl><dt> I </dt><dd> 151 25 </dd></dl>

<dl><dt> II </dt><dd> 166 18 </dd></dl>

<dl><dt> III * </dt><dd> 102 - </dd></dl>

<dl><dt> racemic cis-FTC </dt><dd> 186 22,8 </dd></dl>

<dl><dt> * Form III does not undergo fusion but a solid state transition at approximately 112 ° C, therefore, heat of fusion is not known. </dt>&lt;dd /&gt;</dl>

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The relationship of thermodynamic stability between these forms is represented graphically in Figure 55 14.

Example 8: Solubility

It is evident in Figure 14 that, below 130 ° C, Form I is the most stable phase. Therefore, 60 below 130 ° C, Form I is the least soluble phase. The equilibrium solubility of Form I in water, at 25 ° C, was 0.482 M (119 mg / ml). Compounds that have solubility values ​​greater than 100 mg / ml are considered highly soluble and Form I of (-) - cis-FTC is included in this category. The other forms of (-) - cis-FTC described in this document would have a greater solubility than Form I.

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Example 9: Crystallization studies

The starting material (-) - cis-FTC for all crystallization studies was Form I of (-) - cis-FTC, determined by PXRD. Form I of (-) - cis-FTC was recrystallized from solutions of water, methanol, tetrahydrofuran, ethyl acetate and acetone. All samples from the recrystallization experiments were analyzed by PRDX and DSC. The (-) - cis-FTC crystallized from ethyl acetate and acetone presented endotherms at 151 ° C and 162 ° C, and PXRD diagrams identical to the Form I diagram of (-) - cis-FTC.

A supersaturated solution (0.5 g / ml) of Form I was prepared at 50 ° C. Subsequently, this solution was cooled to room temperature, with stirring, for approximately 2 hours. The precipitated solid 10 was filtered under vacuum and air dried. This solid was analyzed by HPLC, DSC, PXRD and TGA. These analyzes revealed that the solid was a (-) - cis-FTC sesquihydrate. The equilibrium solubility of sesquihydrate, at 25 ° C, is 0.34 M (03 mg / ml). The DSC thermogram of the hydrate is shown in Figure 12. The large endotherm at low temperatures was due to the loss of hydration water from (±) -cis-FTC sesquihydrate, which was confirmed by TGA (Figure 13). The endotherm at 190 ° C was due to the melting of the dehydrated hydrate. Subsequently, an individual melting temperature of approximately 190 ° C was assigned to the dehydrated hydrate, Figure 12. A single PXRD diagram was also obtained for the dehydrated hydrate. (See Figure 11).

PHARMACEUTICAL COMPOSITIONS 20

People suffering from HIV and HBV can be treated by administering to the patient an effective amount of different compounds of the present disclosure (i.e., Forms II and III of (-) - cis-FTC, cis-FTC sesquihydrate racemic and the dehydrated form of the racemic cis-FTC sesquihydrate), or a pharmaceutically acceptable salt thereof, in the presence of a pharmaceutically acceptable carrier or diluent. The active materials are properly formulated for oral administration, in liquid or solid form.

A preferred dose of the compound for HIV or HBV will be in the range of about 1 to 75 mg / kg, preferably 1 to 50 or 20 mg / kg body weight per day, more generally, 0.1 to 30 about 100 mg per kilogram of body weight of the recipient per day. The effective dosage range of pharmaceutically acceptable salts and prodrugs can be calculated based on the weight of the parental nucleoside to be delivered. If the salt exhibits activity on its own, the effective dosage can be estimated as before using the weight of the salt, or by other means known to those skilled in the art. 35

The compound is conveniently administered in any suitable dosage form, which includes, but is not limited to, one containing 7 to 3000 mg, preferably 70 to 1400 mg of active ingredient per dosage unit form. Generally, an oral dosage of 50-1000 mg is convenient. 40

Ideally, to achieve a peak in plasma concentrations, the active compound should be administered at a concentration of about 0.2 to 70 µM, preferably about 1.0 to 10 µM. This could be achieved, for example, by intravenous injection of a solution of 1% to 5% of active ingredient, optionally in saline solution, or by administration as a bolus of the active ingredient.

The concentration of the active compound in the composition of the drug will depend on the rates of absorption, inactivation and excretion of the drug, as well as other factors known to those skilled in the art. It should be taken into account that the dosage values ​​will also vary with the severity of the condition that is to be relieved. It should also be understood that for any given individual, the specific dosage regimens should be adjusted over time according to the individual needs and professional criteria of the person who administers, or supervises the administration, of the compositions, and that The concentration ranges set forth herein are only exemplary and are not intended to limit the scope or practice of the claimed composition. The active ingredient could be administered at once, or it could be divided into several smaller doses for administration at varying time intervals.

A preferred mode of administration of the active compound is orally. Generally, oral compositions will include an inert diluent or an edible carrier. They could be contained in 60 gelatin capsules or be as tablets. For the purpose of oral therapeutic administration, the active ingredient can be incorporated with excipients and used in the form of tablets, pills or capsules. As part of the composition, pharmaceutically compatible binding agents and / or adjuvant materials can be included.

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The tablets, pills, capsules, pills and the like may contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose; a disintegrating agent such as alginic acid, Primogel or corn starch; a lubricant such as magnesium stearate or Sterotes; a sliding agent such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate or orange flavor. When the form of the dosage unit is a capsule, it may contain, in addition to the material of the type indicated, a liquid carrier such as a fatty oil. In addition, the dosage unit forms may contain other different materials that modify the physical form of the dosage unit, for example, sugar coatings, shellac or other enteric agents. 10

The compound can be administered as a component of an elixir, a suspension, a syrup, a wafer, a chewing gum or the like. A syrup could contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorants and flavorings.

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The compound or a pharmaceutically acceptable prodrug or salts thereof may also be mixed with other active materials that do not prevent the desired action, or with materials that supplement the desired action, such as antibiotics, antifungals, anti-inflammatories or other antiviral agents, including others. nucleoside compounds The solutions or suspensions used for parenteral, subcutaneous or topical intradermal application may include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. Parenteral preparation may be included in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

If administered intravenously, preferred vehicles are physiological saline or phosphate buffered saline (PBS).

30

In a preferred embodiment, the active compounds are prepared with vehicles that will protect the compounds against rapid removal from the body, as a contracted release formulation, including implants and microencapsulated delivery systems. Biocompatible, biodegradable polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. The procedures for preparing said formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation.

Also preferred as pharmaceutical carriers are liposomal suspensions (which include liposomes targeting cells infected with monoclonal antibodies against viral antigens). These could be prepared according to procedures known to those skilled in the art, for example, those described in US Pat. No. 4,522,811 (which is incorporated herein by reference in its entirety). For example, liposome formulations could be prepared by dissolving an appropriate lipid (or lipids) (such as stearoyl phosphatidyl ethanolamine, stearoyl phosphatidyl choline, aracadoyl phosphatidyl choline and cholesterol) in an inorganic solvent that subsequently evaporates, leaving a thin film of lipid dry on the surface of the container. Subsequently, an aqueous solution of the active compound or its monophosphate, diphosphate and / or triphosphate derivatives is introduced into the container. Subsequently, the vessel is stirred with a manual circular motion to release the lipid material from the vessel walls and disperse the lipid aggregates, thus forming a liposomal suspension.

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It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope of the invention as defined in the appended claims. It is intended that the specification and examples be considered as exemplary only, indicating the true scope of the invention in the following claims.