NO315555B1 - HIV protease inhibitors, salt and its use, as well as pharmaceutical preparations - Google Patents

Description

The present invention relates to a new series of chemical compounds useful as HIV protease inhibitors. The present origin also relates to the salt and the use of the compound, as well as the pharmaceutical preparation.

Acquired immunodeficiency syndrome (AIDS) is a relatively newly recognized disease or

state. AIDS causes a gradual breakdown of the body's immune system as well as progressive reduction of the central and nervous systems. Since its initial recognition in the early 1980s, AIDS has spread rapidly and has now reached epidemic proportions within a relatively limited segment of the population. Intensive research has led to the discovery of the agent responsible, human T-lymphotrophic retrovirus III (HTLV-III), now commonly referred to as human immunodeficiency virus or HIV.

HIV is a member of the class of viruses known as retroviruses. The retroviral genome consists of RNA that is converted into DNA by reverse transcription. This retroviral DNA is then stably integrated into the host cell's chromosome and by applying replicative processes to the host cells, new retroviral particles are produced and the infections are spread to other cells. HIV appears to have a special affinity for human T-4 lymphocyte cells which play a vital role in the body's immune system. HIV infection of these white blood cells depletes this population of white cells. Finally, the immune system becomes inoperative and ineffective against various opportunistic diseases such as pneumocystis carinii pneumonia, Kaposi's sarcoma and cancer of the lymphatic system.

Despite the fact that the exact mechanism of formation and action of the HIV virus is not known, the identification of vimset has led to some progress in controlling the disease. For example, the drug azidothymidine (AZT) has been found to be effective in inhibiting reverse transcription of the retroviral genome of the HIV virus and thus provides a measure of control but not a cure for patients affected by AIDS. The search continues for drugs that can cure or at least provide an improved target and control of the deadly HIV disease.

Retroviral replication provides for minute post-translational processing of polyproteins. This processing is achieved by the virally encoded HIV protease enzyme.

This provides mature polypeptides which then assist in the formation and function of infectious vims. If this molecular processing is destroyed, the normal production of HIV will end. Inhibitors of HIV protease can therefore act as anti-HIV viral agents.

HIV protease is one of the translated products from the HIV structured protein pol gene. This retroviral protease specifically cleaves other structural polypeptides at specific sites to release these newly activated structural proteins and enzymes, thereby rendering the virus replication competent. Inhibition of the HIV protease by potent compounds can prevent proviral integration of infected T lymphocytes during the early phase of the HIV-1 life cycle and also inhibit viral proteolytic processing during the later stage. In addition, protease inhibitors may have the advantage of being more readily available, living longer in viruses and being less toxic than currently available drugs possibly caused by their specificity for the retroviral protease.

According to the present invention, there is provided a new class of chemical compounds which can inhibit and/or block the activity of HIV protease and which reduce the proliferation of the HIV virus, and pharmaceutical compositions containing these compounds.

The present invention relates to compounds falling under formula (9) below and pharmaceutically acceptable salts and solvates thereof, which inhibit the protease encoded by human immunodeficiency virus (HIV) type 1 (HTV-1) or type 2 (HIV-2). These compounds are useful in the treatment of infection with HTV and the treatment of acquired immunodeficiency syndrome (AIDS). The compounds, their pharmaceutically acceptable salts and pharmaceutical compositions according to the present invention may be used alone or in combination with other antiviral agents, immunomodulators, antibiotics or vaccines. Compounds according to the present invention can also be used as prodrugs. Methods for treating AIDS, methods for treating HIV infection and methods for inhibiting HIV protease are described.

The compounds according to the present invention are characterized by having

formula (9)

where

RerH;

R' is a (C 1 -C 6 )alkyl-OR] group;

where R| is H;

or a pharmaceutically acceptable salt or solvate thereof.

It is further preferred a compound characterized by having the formula:

or a pharmaceutically acceptable salt or solvate thereof.

The present invention further relates to salt according to claim 1, characterized in that it has the formula

The present invention further provides pharmaceutical formulations comprising an effective amount of a compound of formula (9) and a pharmaceutically acceptable carrier.

The present invention further provides a pharmaceutical preparation characterized in that it comprises:

(a) an effective amount of the compound of claim 2; and

(b) a pharmaceutically acceptable carrier therefor.

The present invention further provides for the use of an effective amount of a compound according to claim 1 or a pharmaceutically acceptable prodrug, salt or solvate thereof for the preparation of a pharmaceutical preparation for inhibiting HIV protease.

The present invention further provides for the use of an effective amount of a compound according to claim 2 or a pharmaceutically acceptable salt or solvate thereof for the preparation of a pharmaceutical preparation for inhibiting HIV protease.

The present invention provides new compounds which fall under formula (9),

as described above, which is useful for the treatment of HIV infection and/or AIDS.

Applicants incorporate by reference US Patent No. 5,484,926, US Patent Application Nos. 08/708,411 and 08/708,607 and Japanese Patent Application Nos. JP 95-248183 and JP 95-248184, with the understanding that the definitions of preferences, terms, variables, markers and the like used in each application can only be used in the corresponding description from that application.

Due to the fact that each of the above-mentioned applications incorporated herein by reference was prepared separately, the original applications may in some cases use the same term, marker or variable, but where it is meant to be something else. For example, the variable "X" is used in each application, but each application has its own specific definitions of the substituent or part represented by this variable. It is known to those skilled in the art that terms, markers and variables in each application incorporated by reference are limited only to the description from that application, and may be substituted with other suitable terms, markers and variables or the like representing the particular substituents and moieties . Furthermore, those skilled in the art will recognize that any suitable set of teeth, markers and variables may be used for generic or more

specifically represent the subject matter described in the present application, including terms,

in markers, variables and the like generally applicable in the incorporated descriptions of the above applications and the following description.

The compounds of formula (9) may be prodrugs, which may act to improve the pharmaceutical properties of the compounds, such as pharmacokinetic

properties, e.g. improved bioavailability or solubility. Preparation of the prodrugs can be carried out according to standard procedures known to those skilled in the art.

All temperatures stated herein are in degrees Celsius (°C). All measurement units used herein are

in units of weight with the exception of liquids which are in units of volume.

The term "alkyl" as used herein refers to linear or branched groups having one to six, and most preferably from one to four carbon atoms. The designation "C 1 -C 8 -alkyl"

represents a linear or branched alkyl chain with from one to six carbon atoms. For example, Ci-C6 alkyl groups include methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl,

sec.butyl, t-butyl, pentyl, no-pentyl, hexyl, isohexyl and the like. The term "C 1 -C 6 alkyl" includes within the definition thereof the term "C 1 -C 4 alkyl".

The compounds according to the present invention have at least five asymmetric centers indicated by an asterisk in formula (9) below

As a consequence of these asymmetric centers, the compounds according to the present invention can occur in any possible stereoisomeric forms, and can be used in mixtures of stereoisomers, which can be optically active or racemic, or can be used alone as substantially pure stereoisomers, i.e. at least 95% purity. All the asymmetric forms, individual stereoisomers and combinations thereof fall within the scope of the present invention.

Individual stereoisomers can be prepared from their respective precursors according to the methods described above, by resolution of the racemic mixtures or by separation of the diastereomers. The dissolution can be carried out in the presence of a solvent, by chromatography or by repeated crystallization or by a combination of these techniques known in the art. Further details regarding resolutions can be found in Jacques et al., Enantiomers, Racemates, and Resolutions, John Wiley & Sons 1981.

Preferred of the compounds according to the present invention are substantially pure, ie with 50% purity. It is preferred that the compounds have a purity of at least 75%. It is more preferred that the compounds have a purity of more than 90%. It is even more preferred that the compounds have a purity of at least 95%, more preferably at least 97% and still more preferably at least 99% purity.

As mentioned above, the invention includes pharmaceutically acceptable salts of compounds defined by formula (9). A compound of the present invention may contain sufficient acidic, sufficient basic or both functional groups, and consequently react with any number of inorganic or organic bases, and inorganic and organic acids, to form a pharmaceutically acceptable salt.

The term "pharmaceutically acceptable salt", as used herein, refers to salts of compounds of the above formula which are substantially non-toxic to living organisms. Examples of pharmaceutically acceptable salts include salts prepared by reacting the compounds of the present invention with a mineral or organic acid or an inorganic base. The reactants are generally combined in a common solvent such as diethyl ether or benzene, for acid addition salts or water or alcohols for base addition salts. Salts are normally precipitated out of solution within one hour to about ten days and can be isolated by filtration or other conventional methods. Such salts are known as acid addition and base addition salts.

The acids that can be used to form acid addition salts are inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid and the like, and organic acids such as p-toluenesulfonic acid, methanesulfonic acid, oxalic acid, p-bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid, acetic acid and the like.

Examples of pharmaceutically acceptable salts are sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-l,4-diate, hexyne-l,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, xylene sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, g-hydroxybutyrate, glycolate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate and the like.

Preferred pharmaceutically acceptable acid addition salts are those formed with mineral acids such as hydrochloric acid and hydrobromic acid, and those formed with organic acids such as maleic acid and methanesulfonic acid.

Base addition salts include those derived from inorganic and organic bases, such as ammonium or alkali or alkaline earth metal hydroxides, carbonates, bicarbonates and the like. Bases useful for the preparation of the salts according to the invention accordingly include sodium hydroxide, potassium hydroxide, ammonium hydroxide, potassium carbonate, sodium carbonate, sodium bicarbonate, potassium bicarbonate, calcium hydroxide, calcium carbonate and the like. The potassium and sodium salt forms are particularly preferred.

A "pharmaceutically acceptable solvate" is intended to include a solvate which retains the biological effectiveness and properties of biologically active components of the compounds of formula (9).

Examples of pharmaceutically acceptable solvates include, but are not limited to, compounds of formula (9) in combination with water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid or ethanolamine.

It is to be noted that the specific counterion which forms part of any salt according to the invention is not of a critical nature if the salt as a whole is pharmacologically acceptable and if the counterion does not contribute undesirable qualities to the salt as a whole.

A preferred compound is compound 21

[3S-[2(2S<*>,3S<*>),3-alpha,4a beta, 8a beta]]-N-(1,1-dimethyl-2-hydroxyethyl)decahydro-2-[2-hydroxy -3 - [(3-Hydroxy-2-methylbero^

A method for the preparation of compound 21 is given below. Compound 21 has also been obtained as a metabolite from the plasma of patients administered [3S-(3R,4aR*, 8aR*,2'S*,3'S*)]-2-[2,-hydroxy-3'-phenylthiomethyl-4 '-aza-5'-oxo-5'-(2"-methyl-3"-hydroxyphenyl)pentyl]dicahydroisoquinoline-3-N-t-butylcarboxamide methanesulfonic acid salt, which is described in US Patent No. 5,484,926.

The compounds with formula (9) can be prepared according to the following reaction scheme I.

Reaction scheme I

Compound la, perhydroisoquinoline commercially available from NSC Technologies (Chicago, IL) or Procos SpA (Milan, Italy) is subjected to extended acid hydrolysis in step la to obtain compound 2a. A quantity of inorganic acids which were used in either an aqueous/organic solvent mixture or in water alone at temperatures above 50°C. An example of such an inorganic acid is 6N aqueous HG. Substituents for compound la include the following esters lb, thioesters lc or other amides ld:

where Z, Z1 and Z2 can each independently be alkyl, cycloalkyl, heterocyclic group or aryl.

Compound 2A is then protected by amine nitrogen to obtain compound 2b in step 1b. The protecting group Rp is defined as a suitable conjugating group to avoid unwanted decomposition of activated carboxylate derivatives of compound 2b in step 2. Such protecting groups can usually be of carbamate origin, with a general structure of formula 11:

The identity of R" in formula 11 can be any alkyl, cycloalkyl, aryl or heterocyclic group that can be easily removed in a deprotection step after step 2. Examples of R" include, but are not limited to methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, t-butyl or higher branched or linear alkyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, allyl, phenyl, substituted phenyl, benzyl, substituted benzyl, 9-fluorenylmethyl, 9-anthrylmethyl and higher polycyclic aromatic ring system. The following materials, as defined below, may be obtained from Aldrich Chemical Co. (Sigma Aldrich Fluka):

Such protecting groups can usually be introduced according to an acylation reaction of the corresponding haloformate ester 12a or a dicarbonate 12b: in the presence of a suitable base in typical organic solvents for these types of reactions such as halogenated solvents, ethers and hydrocarbons. Such bases are usually inorganic, such as metal hydroxides, bicarbonates and carbonates or organic bases such as amines such as triethylamine, diethylamine, diethylisopropylamine, 1,8-diazabicyclo[2.2.2]octane (DABCO) or related di- or trialkylamines, as well as amidine bases such as 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and 1,8-diazabicyclo[4.3.0]non-5-ene (DBN). The following materials, as defined below, may be obtained from Aldrich Chemical Co. (Sigma Aldrich Fluka):

These reactions are usually run from below room temperature to about 100°C.

Amide coupling step 2 can be achieved in any way depending on how the carboxyl group is activated. A group J is introduced in step 2 by reaction of carboxylic acid 2b to form activated derivative 2C.

The group J can be any of a number of leaving groups such as alkoxy, hydroxy, halogen, pseudohalogen (including azide, cyanide, isocyanate and isothiocyanate), alkyl or arene sulfonate, aromatic heterocyclic group (bonded through a heteroatom) and N-hydroxyheterocyclic group , including hydroxysuccinimide or hydroxybenzotriazole esters. The following definitions apply to the above designations:

Acyl halides (2c, J = halogen) can be prepared using inorganic halogenating agents such as thionyl chloride or bromide, phosphorus trichloride or bromide, phosphorus pentachloride or bromide or organic agents such as oxalyl chloride or trichloroisocyanuric acid. Esters (2c, J = OR") (R" is defined above) and can be prepared in various ways starting from the acid chloride 2c where J is Cl by combination with the desired alcohol in the presence of an organic or inorganic base previously indicated for the acylation of compound 12a or compound 12b. Alternatively, the ester can be produced by acid-external esterification in the presence of the desired alcohol. Sulfonates (2c, J = OSO2W1, where W[ is alkyl or aryl) are usually formed by reacting carboxylic acid 2b with alkyl or arylsulfonyl chlorides in the presence of an organic amine base such as triethylamine in a non-polar solvent at temperatures below 0°C. Alkyl and arylsulfonyl are defined as follows:

Pseudohalide derivatives according to 2c (J = pseudohalide) are usually prepared from acid halides 2c (J = halogen) by addition of inorganic pseudohalide in the presence of a base. Such bases include, but are not limited to, metal hydroxides, bicarbonates and carbonates or organic bases such as amines such as triethylamine, diethylamine, diethylisopropylamine, 1,8-diazabicyclo[2.2.2]octane (DABCO) or related di- or trialkylamines, as well as amidine bases such as 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and 1,8-diazabicyclo[4.3.0]non-S-ene (DBN). A particularly preferred base is triethylamine. Heteroaromatic derivatives according to 2c are also formed from the acid halides 2c (J = halogen), using the specific heteroaromatic compound in the presence of an amine base in a nonpolar solvent. N-Hydroxyheterocyclic derivatives of 2c can be formed from acid halides as above and can also be formed using alkylcarbodiimides (alkyl-n=C=N-alkyl, where the alkyl groups can be the same or different) or arylcarbodiimides (aryl-N=C= N-aryl, where the aryl groups may be the same or different) and an amine base as condensing agents.

Primary or secondary amine (shown above the arrow in step 2 according to Scheme I) used in the coupling process can incorporate suitable protecting groups, depending on the functionality present in the amine and the coupling method used. The coupling method according to 2c with a primary or secondary amine can be carried out in different ways depending on the identity to J. When a free acid is used (2c, J = OH), the coupling can be accomplished using carbodiimide-based methods using any of the common reagents of this class, including dicyclohexylcarbodiimide or related dialkylcarbodiimides, EDC (salts of l -(3-dimethylaminopropyl)-3-ethylcarbodiimide) or related water-soluble reagents together with an organic amine base in polar organic solvents such as dioxane, DMF, NMP and acetonitrile in the presence of an N-hydroxy heterocyclic compound such as N-hydroxysuccinimide or 3 -hydroxybenzotriazole. Alternatively, haloformate esters, such as 12d, can be used to temporarily activate the acid to provide mixed anhydrides of general formula 2d.

Such haloformate esters are generally as shown in 12d above and include methyl, ethyl, isopropyl, isobutyl, n-butyl, phenyl and related alkyl and aryl chloroformates defined below.

Formula 2d is a possible intermediate in the step from formula 2b to formula 3. Formula 2d is an intermediate, but the process described here results in formula 3, without isolation of formula 2d.

These reactions are usually carried out in a variety of non-polar organic solvents such as halocarbons and ethers such as diethyl ether, methyl t-butyl ether, diisopropyl ether, dioxane and THF at temperatures below 0°C accompanied by an organic amine base such as triethylamine, diethylamine, diethylisopropylamine, DABCO or related di- or trialkylamines, as well as amidine bases such as DBU and DBN.

When J in compound 2c is an alkyl or arene sulfonate (J = OSO2R or OSChAr), the coupling can be carried out in various nonpolar organic solvents such as halocarbons and ethers, such as diethyl ether, methyl t-butyl ether, diisopropyl ether, doxane and THF at temperatures below 0 °C, accompanied by an organic amine base such as triethylamine, diethylamine, diethylisopropylamine, DABCO or related di- or trialkylamines, as well as amidine bases such as DBU and DBN.

When J in compound 2c is a halogen or pseudohalogen, the coupling can be carried out in most common organic solvents such as THF, diethyl ether, dioxane, methyl t-butyl ester or other ethers; acetone, cyclohexanone, methyl isobutyl ketone and other ketones; ethers such as ethyl, methyl and isopropyl acetate; halogenated solvents such as halogenated methanes and ethanes, chlorobenzene and other halogenated benzenes; nitriles such as acetonitrile and propionitrile; lower alcohols such as ethanol, isopropanol, t-butanol and related alcohols; and polar organic solvents such as dimethylformamide, dimethylsulfoxide, N-methyl-2-pyrrolidine and related amide-containing solvents. A base is often used and may be any of a number of inorganic bases such as metal hydroxides, bicarbonates and carbonates or organic bases such as amines such as triethylamine, diethylamine, diethylisopropylamine, DABCO or related di- or trialkylamines, as well as amidine bases such as DBU and DBN.

Those skilled in the art will be able to perform amide coupling step 2 with other possible J groups.

In step 3, removal of the protecting group can be achieved using any standard method for deprotection of a particular class of protecting group. Simple alkyl and substituted alkyl carbamates can be removed with aqueous solutions of the base at temperatures up to 100°C using any of the common inorganic metal hydroxides such as sodium, lithium, potassium or barium hydroxide or hydroxides of other metals in at least stoichiometric amounts. The carbamate protecting groups containing the benzyl groups bound to oxygen can be removed by hydrogenolysis with a palladium or platinum catalyst. Alternatively, aqueous base hydrolysis may be employed at temperatures up to about 100°C using any of the common inorganic metal hydroxides such as sodium, lithium, potassium or barium hydroxide or hydroxides of other metals in at least stoichiometric amounts. A number of anhydrous acids can also be used for deprotection of benzyl-based carbamates, including HCl, HBr and HI. Lewis acids of boron and aluminum such as AICI3, BBr3, BCI3 in non-polar solvents are also effective. Certain substituted benzyl, aryl or alkyl groups where the specific substitution pattern is chosen for its ability to be removed under specific conditions may also be used. For example, the 2-trimethylsilylethylcarbonyl group (Teoc) is a protecting group designed to take advantage of the specific reactivity of the 2-trimethylsilylethyl group in the deprotection process. 2-trimethylsilylethylcarbonyl chloride can be used to protect the amine nitrogen and can later be removed using fluoride ion sources such as HF or tetraalkylammonium fluoride salts.

In step 4, the perhydroisoquinoline moiety according to formula 4 is coupled to chloroalcohol (compound 5, scheme I) via an epoxide intermediate (13) formed via base-induced closure of adjacent chlorohydrin functionality.

Compound 5 is manufactured by Kaneka industries, Japan. Several close-open procedures starting from compound 5 -» compound 13 -> compound 6 can be used. Epoxide 13 may be isolated or may be reacted with 4 added either after the formation of 13 or 4 may be present from the beginning of the sequence. Epoxide 13 can be formed using inorganic bases such as metal hydroxides, carbonates and bicarbonates in solvents such as alcohols such as methanol ethanol or isopropyl alcohol, ethers such as THF and dioxane or mixtures of the two. Epoxide can also be formed in a 2-phase solvent system consisting of water and a halocarbon solvent such as dichloromethane together with the base. A phase transfer catalyst such as a tetraalkylammonium salt can be used to facilitate the process. The process of opening epoxide 13 with compound 4 is accomplished in alcohol solvents or mixtures of an alcohol and another solvent which may be an ether or dipolar aprotic solvent such as dimethylformamide or dimethylsulfoxide. Opening of epoxide 13 with compound 4 to provide compound 6 is preferably carried out over a period of 2-7 hours at 50-60°C.

In step 5, the carbobenzyloxy group can be removed to provide free amine 7. This can be done using HBr in acetic acid using carbon solvents such as halocarbons. It can also be carried out using halides of boron such as BBtj and BCI3 or alkyl-substituted boron halides such as dimethyl boron bromide in halocarbon solvents such as chloroform and dichloromethane at temperatures varying from 0°C up to room temperature. Alternatively, the carbobenzyloxy group can be removed by hydrolysis using aqueous/alcoholic solutions of metal hydroxides such as barium, sodium, lithium or potassium hydroxide at temperatures above room temperature for several hours.

Step 6a is the coupling of benzoic acid derivatives of formula 8 to provide 9a. In formula 8, Q can be a leaving group. Q may be any of the leaving groups discussed above for group J. The compounds of formula 8 where Q = OH or Cl are commercially available from EMS Dottikon, Lenzburg, Switzerland and Sugai Chemical Industries, Ltd. in Japan. The coupling can be carried out in different ways depending on the identity of Q. When a free acid is used (Q = OH) the coupling can be carried out using carbodiimide based methods using any of the usual reagents from this class including dicyclohexylcarbodiimide or related dialkylcarbodiimides, EDC (salts of l-(3-dimethylaminopropyl)-3-ethylcarbodiimide or related water-soluble reagents together with an organic amine base in polar organic solvents such as dioxane, DMF, NMP and acetonitrile in the presence of an N-hydroxy heterocyclic group including N- hydroxysuccinimide or 3-hydroxybenzotriazole. When Q = a halogen or pseudohalogen, the coupling can be carried out in the most common organic solvents such as THF, diethyl ether, dioxane, methyl t-butyl ether or other ethers; acetone, cyclohexanone, methyl isobutyl ketone and other ketones; esters such such as ethyl, methyl and isopropyl acetate, halogenated solvents such as halogenated methanes and ethanes, k chlorobenzene and other halogenated bases, nitriles such as acetonitrile and propionitrile; lower alcohols such as ethanol, isopropanol, t-butanol and related alcohols, and polar organic solvents such as dimethylformamide, dimethylsulfoxide, N-methyl-2-pyrrolidone and related amide-containing solvents. A base is often used and can be any of a number of inorganic bases such as metal hydroxides, bicarbonates and carbonates or organic bases such as amines such as triethylamine, diethylamine, diethylisopropylamine, DABCO or related di- or trialkylamines, as well as amidine bases such as DBU and DBN .

Acetate removal is achieved in step 6b with aqueous or alcoholic solutions of inorganic bases such as metal hydroxides, carbonates and bicarbonates at room temperatures up to 100°C. If there is a protected functionality on the carboxamide group attached to the perhydroisoquinoline ring system, it is best removed at this point (during or after step 6b). The nature of this step depends on the exact identity of the protecting group.

A preferred method for achieving the entire process shown in Scheme I is shown in Scheme II.

Cbz-protected amino acid 15 was coupled with amine 22 to provide amine 16. The Cbz group was removed by hydrogenation to provide 17.

This was coupled with chloroalcohol via the epoxide using the in situ procedure to provide adduct 18. Conventional deprotection with base and coupling of free primary amine with acid chloride 20 gave rise to amide 21. Details of this procedure are given below in Examples 1 A to F .The letters A to F in Form II correspond to those in Examples 1 A to F below.

The following examples illustrate the aspects according to the invention. The abbreviations for the terms melting point, nuclear magnetic resonance spectra, electron impact mass spectra, field-desorption mass spectra, solid atom bombardment mass spectra, infrared spectra, ultraviolet spectra, elemental analysis, high performance liquid chromatography and thin layer chromatography are respectively m.p., NMR, EIMS, MS(FD ), MS(FAB), IR, UV, analysis, HPLC and TLC. In addition, the absorption maxima listed for the IR spectra are those of interest, and not all observed maxima.

In the context of the NMR spectra, the following abbreviations are used: singlet (s), doublet (d), doublet of doublets (dd), triplet (t), quartet (q), multiplet (m), doublet of multidoublets (dm), wide singlet (br.s), wide doublet (br.d), wide triplet (br.t) and wide multiplet (br.m). J indicates the coupling constant in Hertz (Hz). Unless otherwise stated, NMR data refer to the free base of the compound in question.

NMR spectra were obtained on a General Electric QE-300 300MHz instrument. Chemical shifts are expressed in 5-values in ppm. Mass spectra were obtained on a VG ZAB-3 spectrometer at the Scripps Research Institute, La Jolla, Ca. Infrared spectra were recorded on a Midac Corporation spectrometer. The UV spectra were obtained on a Varian Cary 3E instrument. Thin layer chromatography was performed using silica plates available from E. Merck. Melting points were measured on a Mettler FP62 instrument and uncorrected.

Example 1

Process for the preparation of amide of formula 21

[3S-[2(2S<*>,3S<*>),3 alpha,4a beta,8abeta]]-N-(1,1-dimethyl-2-hydroxyethyl)decahydro-2-[2-hydroxy-3 -[(3-hydroxy-2-methylbenzoyl)amino]-4-(phenylthio)butyl]-3-isoquinolinecarboxaniide

A. Perhydrisoquinoline (26.4 g, 111 mmol) (commercially available from NSC Technologies (Chicago, IL) or Procos SpA (Milan, Italy)) was suspended in water (200 mL) and concentrated aqueous HCl (200 mL). This mixture was heated to reflux and stirred for 3 days, after which it went into solution. The solvents were removed under reduced pressure to provide a pale yellow solid. The solid was slurried in 2-propanol (200 mL) and filtered. The filtrate was evaporated under reduced pressure to an oil. EtOAc (100 mL) and water (100 mL) were added and the pH of the solution was adjusted to 8.0 by addition of 2N aqueous KOH. Benzyl chloroformate (15.8 mL, 111 mmol) was added dropwise over 30 min and the pH was maintained between 7 and 8 by addition of 2N aqueous KOH. The mixture was stirred at room temperature for 18 hours. EtOAc (200 mL) was added and the organic layer was washed with 1N aqueous HCl (100 mL), and brine (100 mL). The organic layer was dried (MgSO 4 ), filtered and evaporated under reduced pressure to an oil. The product was purified by silica gel chromatography, eluting with 1:1 40-60 petroleum ether/EtOAc followed by 100% EtOAc. Fractional product was collected and evaporated under reduced pressure to provide compound 15 (11.3 g, 32%) as a colorless oil: <!>H NMR (300 MHz, CDCl 3 ) δ 7.43-7.28 (m , 5 H), 5.17 (br s, 2 H), 4.76 (m, 1 H), 3.79 (m, 1 H), 3.33 (m, 1 H), 2.19 ( m, 1H), 1.96 (m, 1H), 1.88-1.15 (m, 10H).

B. 1-Hydroxybenzotriazole (4.2 g, 31.4 mmol) and EDC (6.0 g, 31.4 mmol) were added to a solution of acid 15 (8.3 g, 26.2 mmol) in DMF (128 mL) at room temperature. The mixture was heated at 80°C for 10 minutes. 1,1-dimethyl-2-trimethylsilyloxyethylamine (5.1 g, 31.4 mmol, prepared from 1,1-dimethyl-2-hydroxyethylamine (Aldrich Chemical Co.) and hexamethyldisilazane (Aldrich Chemical Co.)) by heating the mixture under reflux for several hours followed by evaporation of the volatile components were added and the solution was heated at 80°C for 17 hours. The yellow solution was poured into EtOAc (250 mL) and 2N aqueous HCl (250 mL). After stirring for 10 min, EtOAc (750 mL) was added and the mixture was washed with H 2 O (3 x 500 mL) and brine (1 x 250 mL). Combined aqueous layers were extracted with EtOAc (1 x 250 mL). Combined organic layers were dried (Na 2 SO 4 ) and purified by flash chromatography (50/50 EtOAc/hexanes) to afford compound 16 as a colorless oil (7.9 g, 78%): <]>H NMR (300 MHz, CD3OD) 6 7.36 (m, 5 H), 5.20 (d, J = 8.1 Hz, 1 H), 5.10 (m, 1 H), 4.53 (m, 1 H), 3.78 (dd, J = 13.2,4.4 Hz, 1 H), 3.60 (m, 2 H), 3.48 (d, J = 10.7 Hz, 1 H), 2, 15-1.25 (m, 12 H), 1.31 (s, 3 H), 1.29 (s, 3 H).

C. A mixture of carbamate 16 (7.9 g, 20.4 mmol) and 5% palladium-on-carbon (Pd/C)

(1.6 g) was hydrogenated at 50 psi H2 in absolute EtOH (100 mL) at room temperature for 18 h. The mixture was filtered through celite and evaporated in vacuo to provide amine 17 as a white, crystalline solid: 1 H NMR (300 MHz, CD 3 OD) δ 3.63 (q, J = 7.0 Hz, 2 H), 3 .34 (m, 1 H), 3.27 (dd, J = 11.8, 3.3 Hz, 1 H), 2.91 (m, 1 H), 2.02-1.15 (m, 12H), 1.32 (s, 3H), 1.31 (s, 3H).

D. Aqueous 10.2N NaOH (2.4 mL, 24.5 mmol) was added to a warm (27°C) suspension of chloroalcohol (obtained from Kaneka Industries in Japan) (10.4 g, 28.6 mmol) in isopropanol (IPA)

(104 mL) with mechanical stirring. After 1 hour, 1N aqueous HC1 in IPA (prepared by adding 1 mL concentrated aqueous HC1 to 12 mL IPA) approximately (about 1 mL) was added to neutralize (pH = 7). Amine 17 (5.2 g, 20.4 mmol) was added as a solution in IPA (50 mL) and the thin suspension was heated at 60 °C for 10 h. IPA was removed in vacuo. The residue was diluted with EtOAc (150 mL) and washed with H 2 O (2 x 50 mL), saturated aqueous NaHCO 3 (1 x 50 mL), and brine (1 x 50 mL). Combined aqueous layers were extracted with EtOAc (1 x 25 mL).

Combined organic layers were dried (Na 2 SO 4 ) and purified by flash chromatography (75/25 EtOAc/hexanes, then EtOAc) to afford compound 18 as a white solid (8.98 g, 76%): 1 H NMR ( 300 MHz, CD3OD) 8 7.33 (m, 10 H), 5.08 (AB, Jab = 12.2 Hz, Auab = 12.1 Hz, 2 H), 3.96 (m, 2 H), 3.56 (q, J = 7.3 Hz, 2 H), 3.50 (m, 1 H), 3.20 (dd, J = 13.6,9.2 Hz, 1 H), 3, 03 (m, 1 H), 2.64 (m, 2 H), 2.20-1.20 (m, 14 H), 1.28 (s, 6 H).

E. 50% aqueous NaOH (2.7 g, 1.8 mL, 33.6 mmol) was added to a suspension of carbamate 18 (6.75 g, 11.6 mmol) in IPA (34 mL) at room temperature. The mixture was heated under reflux for 12 hours. After cooling to room temperature, the mixture was diluted with methyl t-butyl ether (MTBE) (600 mL) and washed with H 2 O (2 x 250 mL) and brine (1 x 125 mL). Combined aqueous layers were extracted with MTBE (1 x 150 mL). Combined organic layers were dried (Na 2 SO 4 ) and evaporated in vacuo to provide a mixture of compound 19 and benzyl alcohol as an oily white solid: <*>H NMR (300 MHz, CD 3 OD) δ 7.34 (m, 10 H ), 4.63 (s, 2 H), 3.81 (m, 1 H), 3.58 (m, 3 H), 3.03-2.60 (m, 5 H), 2.17 ( m, 1H), 2.05 (m, 1H), 1.87-1.05 (m, 12H), 1.30 (s, 3H), 1.28 (s, 3H).

F. Triethylamine (3.2 g, 4.3 mL, 31.2 mmol) was added to a solution of the mixture of amine 19 (4.7 g, 10.4 mmol theoretical from 18 ) and benzyl alcohol in EtOH (23 mL) at room temperature. A solution of 3-acetoxy-2-methylbenzoyl chloride (20) (obtained according to the method disclosed in US Patent Application No. 08/708,411, filed September 5, 1996, which is specifically incorporated herein by reference) (2.4 g, 11.5 mmol) in THF (4 mL) was added. After 2 h, 50% aqueous NaOH (4.1 g, 2.8 mL, 52.2 mmol) was added and the mixture was heated under reflux for 1 h. After cooling to room temperature, the mixture was neutralized to pH = 7 with 2N aqueous HCl (26 mL). This mixture was diluted with EtOAc (500 mL) and washed with H 2 O (1 x 250 mL), saturated aqueous NaHCO 3 (2 x 250 mL), H 2 O (1 x 250 mL) and brine (81 x 125 mL). The organic layer was dried (Na 2 SO 4 ) and purified by flash chromatography (75/25 EtOAc/hexanes) to afford amide 21 as a white foam (1173-57A, 1.39 g, 23%). <]>H NMR indicated the presence of 11 wt% EtOAc which could not be removed in vacuo.

Analyze:

1H NMR (300 MHz, CD3OD) δ 7.53 (d, J = 7.3 Hz, 2H), 7.32 (t, J = 7.0 Hz, 2H), 7.20 (t, J 7.3 Hz, 1 H), 7.06 (t, J = 8.1 Hz, 1 H), 6.92 (d, J = 8.1 Hz, 1H), 6.83 (d, J = 8.1Hz, 1H), 4.42

(m, 1 H), 4.08 (m, 1 H), 3.61 (dd, J = 13.6,4.0 Hz, 1 H), 3.45 (AB Jab = 11.0 Hz, Au ab = 18.0 Hz, 2 H), 3.29 (dd, J = 13.6,10.3 Hz, 1 H), 3.10 (m, 1 H), 2.66 (m, 2 H), 2.28 (s, 3 H), 2.22 (m, 2 H), 2.04 (m, 1 H), 1.86-1.20 (m, 11 H), 1.19 (p, 3H), 1.18 (p, 3H).

<13>C NMR (75.5 MHz, CD3OD) S 175.7, 172.5, 155.9, 138.8, 136.7, 129.8, 128.9, 126.3, 126.0, 122.4, 118.4, 115.9, 70,3,69,9,68,2,59,3,58,8,54,9, 53,0,36,5,34,2,34,1,31,1 30,7, 26,4 , 26.0, 23.1, 23.0, 20.8, 12.1, .

Example 2

HIV protease inhibition activity and anti-HIV activity in the cell culture of compound 21.

Tight binding kinetics analysis was used to determine the magnitude of the Ki value of compound 21. Kj = 5.6 ± 0.91 nM.

Methods

Expression of HIV-1 protease

The HIV-1 protease gene was isolated from the viral strain UIB (Råtner, L. et al., Nature, 316, 227-284 (1985)). To increase the stability of purified protease (Rose, J.R. et al., J. Biol. Chem., 268,11939-11945 (1993)), the glutamine residue at position 7 (Q7) was mutated to serine (S) by replacement of 33 the base pair segment between the Ndel and BstEII sites of the protease gene sequence with synthetic oligonucleotides coding for the Q7S mutation. The modified gene sequence was inserted into plasmid vector pGZ (Menge, K. L. et al., Biochemistry, 34:15934-15942 (1995) under the control phage T7 promoter. The resulting construct pGZ/HJM9Q7S#9, was transferred into E. coii strain BL21 (DE3) obtained from Novagen, Inc.

Expression of HIV-1 PR: Cuttings were grown in 2YT medium (1.6% trypticase peptone, 1% yeast extract, 0.5% NaCl at initial pH 7.5) containing 200 u.g/1 ampicillin in 1001 fermentor (Biolafitte SA) at 37°C for 5 h and then induced by the addition of 1 mM IPTG (isopropyl-(3-D-thiogalactopyranoside). The temperature of the culture during induction was increased to 42°C to increase the accumulations of recombinant HIV-1 protease as insoluble inclusion bodies After 2 hours at 42°C, cells were harvested by cross-flow filtration using Pellicon 0.1 µm WPP000C5 cartridge #10 (Millipore) and the cell paste was stored frozen at -70°C.

Purification of recombinant HIV-1 protease: Unless otherwise stated, all steps were performed at 4°C. Protein concentrations were determined using BioRad protein assay solution with bovine serum albumin (BioRad, Richmond, CA) as a standard. The chromatography steps and purity of HIV PR were analyzed by sodium dodecylsulphur polyacrylamide gel electrophoresis (SDS-PAGE). Final purity of HTV-PR was

>98%. Typical final yield from each 1001 culture of ~120 mg.

Cell paste from 1001 culture was resuspended in 300 ml lysis buffer (50 mM Tris-Cl pH 8, 0, 25 mM NaCl, 20 mM 2-mercaptoethanol) and microfluidized in a Microfluidics Corporation fluidizer of 22,000 psi. Crude cell lysate was prepared by centrifugation at 14,000 rpm for 20 minutes. HIV PR was mainly found in the pellet in the form of inclusion bodies. The inclusion bodies were then washed several times in lysis buffer additionally containing 0.1% trition-X100 and 1 M urea, and after each washing procedure the inclusion bodies were pelleted by centrifugation at 5,000 rpm for 20 minutes. Purified inclusion bodies were dissolved in buffer containing 50 mM Tris-Cl, pH 8.0, 25 mM NaCl, 20 mM 2-mercaptoethanol and 8 M urea. The solution was prepared by centrifugation at 14,000 rpm and applied at room temperature to a 300 ml Fast Flow Q-Sepharose column (Pharmacia, Piscataway, NJ) equilibrated with the same buffer. Under these conditions, HIV PR was not bound to the column and substantially pure enzyme was found in the flow-through fractions. To renature the protein, fractions from the Fast Flow Q-Sepharose column were dialyzed against three buffer changes containing 25 mM NaH2PO4 pH 7.0, 25 mM NaCl, 10 mM DTT and 10% glycerol. After refolding, small amounts of precipitated material were removed by centrifugation and the resulting enzyme preparation was concentrated, dialyzed against 0.5 M NaCl, 50 mM MES pH 5.6, 10 mM DTT, frozen in small aliquots at ~2 mg/ml and stored at -70°C.

Tight-binding kinetics analysis and assays

The proteolytic activity of purified HIV-1 protease was measured using a modified chromogenic assay developed by Richards et al. (Richards, A.D. et al. J. Biol. Chem., 256, 773-7736 (1990)). The synthetic peptide His-Lys-Ala-Arg-Val-Uu-Phe(paraNO2>-Glu-Ala-Nle-Ser-NH2 (American Peptide Company) (Nie is norleucine) was used as a substrate. The assay was performed in 0 .5 M NaCl, 50 mM MES pH 5.6, 5 mM DDT and 2% DMSO at 37° C. Cleavage of the cleavable bond between leucine and paranitro-phenylalanine (Phe para-NO2) was analyzed by spectrophotometric recording of the reduction of the absorbance at 305 nm. Initial rate was determined as the rate of decrease of absorbance during the first 100 seconds of the enzymatic reaction. Under these conditions and using Q7S HIV-1 protease, the Michaelis constant (Km) for this substrate is 59 ± 17 uM.

To determine the inhibition of compound 21, a saturating concentration of substrate 200 µM was used. Between 13 and 20 concentrations of inhibitors were assessed and the reaction rate was measured at each concentration as described above. Apparent Ki (Ki app), given above, was determined by data-assisted non-linear least squares fitting of the data to the "tight binding" equation of Morrison (Morrison, J.F., Biochem. Biophys. Acta, 185, 269-286(1963)).

Example 3

Antiviral activity of compound 21 against HIV-1 in cell culture

Cells and virus strains:

CEM-SS and MT-2 human T-cell lines and HIV-1 strains RF and UIB were obtained from the AIDS Research and Reference Program, Division of AIDS, NIAJD, and NEH.

Cell protection assays:

Inhibitory effects of each agent on HIV-1 replication were measured according to the MMTT color reduction method (Alley, M.C. et al., Cancer Res. 48: 589-601 (1988)). The compounds were dissolved in DMSO at a concentration of 40 mg/ml and then diluted 1:200 in culture medium (RPMI, supplemented with 10% fetal bovine serum). From each diluted stock medium, 100 µl was added to a 96-well dish and serial half-log dilutions were made. In separate tubes, MT-2 cells and CEM-SS cells were infected with HIV-1 UIB or HIV-1 RF at a multiplicity of infection (m.o.i.) of 0.01 and 0.03, respectively. After a 4-hour adsorption period, 100 µl of infected or uninfected cells were added to the wells of the drug-containing dish to provide a final concentration of 1 x 10<4> cells/well. Six days (CEM-SS cells) or 7 days (MT-2 cells) later, MTT (5 mg/ml) was added to the test dishes and the amount of formazan produced was quantified spectrophotometrically at 570 nm. Data were expressed as percentage of formazan produced in drug-treated cells compared to formazan produced in the wells of uninfected, drug-free cells. The ED50 was calculated as the concentration of drug that increased the percentage of pharmazone production in infected, drug-treated cells to 50% of that produced by uninfected, drug-free cells. The cytotoxicity (TC50) was calculated as the concentration of drug that increased the percentage of formazan produced in uninfected, drug-treated cells to 50% of that produced in uninfected, drug-free cells. Therapeutic index (TI) was calculated by dividing the cytotoxicity (TC30) by the anti-viral efficacy (ED50).

As indicated above, the compounds of the present invention are useful for inhibiting HIV protease, which is an enzyme associated with viral component production and assembly. One embodiment of the present invention is a method for treating HIV infection comprising administering to a host or patient, such as a primate, an effective amount of a compound of formula (9) or a pharmaceutically acceptable salt thereof. Another embodiment according to the present invention is a method for treating AIDS comprising administering to a host or patient an effective amount of a compound of formula (9) or a pharmaceutically acceptable salt thereof. A further embodiment according to the present invention is a method for inhibiting HIV protease comprising administering to an HIV-infected cell or a host or patient, such as a primate, infected with HIV, an effective amount of a compound of formula (1) or a pharmaceutically acceptable salt thereof.

The term "effective amount" means an amount of a compound of formula (9) or its pharmaceutically acceptable salt that effectively inhibits HIV protease mediated viral component production and assembly. The specific dose of the compound administered according to this invention to achieve therapeutic or inhibitory effects will, of course, be determined based on the particular circumstances surrounding the case, including, for example, the compound being administered, the route of administration, the condition being treated, and the individual the host or patient being treated. An example of a daily dose (administered in single or divided doses) contains a dosage level of from about 0.01 mg/kg to about 50 mg/kg body weight of a compound of the invention. Preferred daily doses are generally from about 0.05 mg/kg to about 40 mg/kg and more preferred are from about 1.0 mg/kg to about 30 mg/kg.

The compounds of the invention can be administered by various routes, including orally, rectally, transdermally, subcutaneously, intravenously, intramuscularly and by the intranasal route. The compounds according to the invention are preferably formulated before administration. Another embodiment according to the present invention is therefore a pharmaceutical mixture or formulation comprising an effective amount of a compound of formula (9) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, such as a diluent or an excipient.

The active ingredient preferably comprises from 0.1% to 99.9% by weight of the formulation. By "pharmaceutically acceptable" is meant that the carrier, such as the diluent or excipient, is compatible with the other ingredients of the formulation and not harmful to the host or patient.

Pharmaceutical formulations can be prepared from the compounds according to the invention by means of known methods using known and easily available ingredients. When forming the mixtures according to the present invention, the active ingredient will usually be mixed together with a carrier, or diluted with a carrier, or enclosed in a carrier, which may be in the form of a capsule, pad, paper or other suitable container. When the carrier acts as a diluent, it may be a solid, semi-solid or liquid material which acts as a carrier, excipient or medium for the active ingredient. The mixtures can therefore be in the form of tablets, pills, powders, lozenges, pads, elixirs, suspensions, emulsions, solutions, syrups, aerosols (such as a solid or liquid medium), ointments (containing e.g. up to 10% by weight of the active compound), soft and hard gelatin capsules, suppositories, sterile injectable solutions, sterile packed powders and the like.

The following formulation examples illustrate the scope of the invention. The term "active ingredient" represents a compound of formula (9) or a pharmaceutically acceptable salt thereof.

Formulation 1

Hard gelatin capsules are prepared using the following ingredients:

Formulation 2

A tablet is prepared using the following ingredients:

The compounds are mixed and compressed to form tablets each weighing 665 mg.

Formulation 3

An aerosol solution is formed containing the following components:

The active compound is mixed together with ethanol and the mixture added to a portion of propellant 22, cooled to -30°C and transferred to a filling device. The required amount is then added to a stainless steel container and diluted with the remaining propellant. The valve assemblies are then placed on the container.

Formulation 4

Tablets, each containing 60 mg of active ingredient are formed as follows:

The active ingredient, starch and cellulose are passed through a No. 45 mesh US sieve and thoroughly mixed. Aqueous solution containing polyvinylpyrrolidone is mixed with resulting powder and the mixture is then passed through a No. 14 mesh US sieve. Granules produced in this way are dried at 50°C and passed through a No. 18 mesh US sieve. Sodium carboxymethyl starch, magnesium stearate and talc, previously passed through a No. 60 mesh US sieve are then added to the granules which after mixing are compacted on a

tablet machine to provide tablets each weighing 150 mg.

Formulation 5

Capsules, each containing 80 mg of active ingredient, are formed as follows:

Active ingredient, cellulose, starch and magnesium stearate are mixed, passed through a No. 45 mesh US sieve and filled into hard gelatin capsules in quantities of 200 mg.

Formulation 6

Suppositories, containing 225 mg of active ingredient are formed as follows:

The active ingredient is passed through a No. 60 mesh US sieve and suspended in saturated fatty acid glycerides pre-melted using minimal heat required. The mixture is then poured into a suppository form of nominal 2 g capacity and cooled.

Formulation 7

Suspensions, each containing 50 mg of active ingredient per 5 ml dose are formed as follows:

Active ingredient is passed through a No. 45 mesh US sieve and mixed with sodium carboxymethyl cellulose and syrup to form a smooth paste. Benzoic acid solution, taste and color are diluted with a portion of water and added with stirring. Sufficient water is then added to form the required volume.

Formulation 8

An intravenous formulation is formed as follows:

A solution of the above ingredients is generally administered intravenously to a subject at a rate of 1 ml per minute.

Formulation 9

A tablet is formed using the ingredients below:

Claims (31)

1. Compound, characterized in that it has formula (9): where R is H; R' is a (C 1 -C 6 )alkyl-OR 1 group; where Ki is H; or a pharmaceutically acceptable salt or solvate thereof. 2. Compound, characterized in that it has the formula: or a pharmaceutically acceptable salt or solvate thereof. 3. Salt according to claim 1, characterized in that it has the formula 4. Pharmaceutical preparation, characterized in that it comprises: (a) an effective amount of compound according to claim 1; and (b) a pharmaceutically acceptable carrier therefor. 5. Pharmaceutical preparation, characterized in that it comprises: (a) an effective amount of the compound according to claim 2; and (b) a pharmaceutically acceptable carrier therefor. 6. Use of an effective amount of a compound according to claim 1 or a pharmaceutically acceptable salt or solvate thereof for the manufacture of a pharmaceutical preparation for inhibiting HIV protease. 7. Use of an effective amount of a compound according to claim 2 or a pharmaceutically acceptable salt or solvate thereof for the production of a pharmaceutical preparation for inhibiting HIV protease. 8. Compound according to claim 1, characterized in that it has a purity of more than 90%. 9. Compound according to claim 1, characterized in that it has a purity of more than 95%. 10. Compound according to claim 1, characterized in that it has a purity of at least 97%. 11. Compound according to claim 1, characterized in that it has a purity of at least 99%. 12. Compound according to claim 2, characterized in that it has a purity of more than 90%. 13. Compound according to claim 2, characterized in that it has a purity of at least 95%. 14. Compound according to claim 2, characterized in that it has a purity of at least 97%. 15. Compound according to claim 2, characterized in that it has a purity of at least 99%. 16. Pharmaceutical mixture according to claim 4, characterized in that the compound has a purity of more than 90%. 17. Pharmaceutical mixture according to claim 4, characterized in that the compound has a purity of at least 95%. 18. Pharmaceutical mixture according to claim 4, characterized in that the compound has a purity of at least 97%. 19. Pharmaceutical mixture according to claim 4, characterized in that the compound has a purity of at least 99%. 20. Pharmaceutical mixture according to claim 5, characterized in that the compound has a purity of more than 90%. 21. Pharmaceutical mixture according to claim 5, characterized in that the compound has a purity of at least 95%. 22. Pharmaceutical mixture according to claim 5, characterized in that the compound has a purity of at least 97%. 23. Pharmaceutical mixture according to claim 5, characterized in that the compound has a purity of at least 99%. 24. Use according to claim 6, in that the compound has a purity of more than 90%. 25. Use according to claim 6, in that the compound has a purity of at least 95%. 26. Use according to claim 6, in that the compound has a purity of at least 97%. 27. Use according to claim 6, in that the compound has a purity of at least 99%. 28. Use according to claim 7, in that the compound has a purity of more than 90%. 29. Use according to claim 7, in that the compound has a purity of at least 95%. 30. Use according to claim 7, in that the compound has a purity of at least 97%. 31. Use according to claim 7, in that the compound has a purity of at least 99%.