EA002378B1 - Hiv protease inhibitors - Google Patents

Abstract

1. A compound of the formula: or a pharmaceutically acceptable salt or solvate thereof. 2. A compound according to Claim 1, which has a purity of more than 90%. 3. A compound according to Claim 1, which has a purity of at least 95%. 4. A compound according to Claim 1, which has a purity of at least 97%. 5. A compound according to Claim 1, which has a purity of at least 99%. 6. A pharmaceutical composition for treating HIV infection, comprising effective amount of the compound or salt or solvate thereof according to Claim 1 and a pharmaceutically acceptable carrier. 7. A pharmaceutical composition according to Claim 6 wherein the compound or salt or solvate thereof has a purity of more than 90%. 8. A pharmaceutical composition according to Claim 6 wherein the compound or salt or solvate thereof has a purity of at least 95%. 9. A pharmaceutical composition according to Claim 6 wherein the compound or salt or solvate thereof has a purity of at least 97%. 10. A pharmaceutical composition according to Claim 6 wherein the compound or salt or solvate thereof has a purity of at least 99%.

Description

BACKGROUND AND SUMMARY OF THE INVENTION

This invention relates to a new series of chemical compounds used as HIV protease inhibitors and to the use of such compounds as antiviral agents.

Acquired Immunodeficiency Syndrome (AIDS) is a relatively recently discovered disease or condition. AIDS causes a sequential destruction of the immune system, as well as a progressive dysfunction of the central and peripheral nervous system. Since the description of the first cases of the disease in the early 1980s, HIV infection has spread widely and to date has reached epidemiological values in relatively limited segments of the population. Intensive research has led to the discovery of the causative agent, human T-lymphotropic retrovirus III (NTL-III), which is now commonly referred to as human immunodeficiency virus, or HIV.

HIV is a member of the class of viruses known as retroviruses. The genome of a retrovirus consists of RNA, which is transcribed into DNA through reverse transcription. Then, this retrovirus DNA is stably integrated into the chromosome of the host cell using the replicative processes of the host cell, producing new retroviral particles and further infection of other cells. It seems possible that HIV has a particular affinity for T-4 human lymphocytes, which play a vital role in the immune system. HIV infection of these white blood cells depletes this cell population. Ultimately, the immune system becomes ineffective and ineffective against various opportunistic diseases, such as, among others, pneumonia caused by Rheitos lupus noodles, Kaposi's sarcoma and malignant tumors of the lymphatic system.

Despite the fact that the exact mechanism of the formation and activity of HIV has not been studied, the identification of the virus has led to some progress in the treatment of the disease. For example, it was found that the drug azidothymidine (AZT) is effective in inhibiting the reverse transcription of the HIV retroviral genome, thus providing partial treatment, although not a complete cure for AIDS patients. The search continues for medicines that can cure or at least provide more effective treatment for the still deadly HIV infection.

Retrovirus replication definitely reflects post-translational processing of polypeptides. This processing is carried out by a virus encoded protease enzyme

HIV This leads to the formation of mature polypeptides that sequentially attach to the formation and functioning of the infectious virus. If this molecular processing is inhibited, normal HIV production will be stopped. Thus, HIV protease inhibitors can function as antiviral anti-HIV agents.

HIV protease is one of the translation products of the gene p1 of the HIV structural protein. This retroviral protease specifically cleaves other structural polypeptides at individual sites to release these activated structural proteins and enzymes, thereby imparting replication ability to the virion. Inhibition of HIV protease by potent compounds, as such, can prevent provirus integration in infected Tlymphocytes during the early phase of the HIV-1 life cycle, as well as inhibit the proteolytic processing of the virus from a late stage. In addition, protease inhibitors may have the advantage of greater readability, longer viral life and lower toxicity than currently available drugs, possibly due to their specificity for the retrovirus protease.

In accordance with this invention, which relates to a new class of compounds capable of inhibiting and / or blocking the activity of HIV protease, which stop the proliferation of HIV, pharmaceutical compositions containing these compounds, and the use of these compounds as HIV protease inhibitors.

The present invention relates to compounds defined by formula 9 below and pharmaceutically acceptable salts, prodrugs and solvates thereof that inhibit a protease encoded by human immunodeficiency virus (HIV) type 1 (HIV1) or type 2 (HIV-2). These compounds are useful for treating HIV infection and for treating acquired immunodeficiency syndrome (AIDS). The compounds, their pharmaceutically acceptable salts and pharmaceutical compositions of the present invention can be used alone or in combination with other antiviral drugs, immunomodulators, antibiotics or vaccines. The compounds of the present invention can also be used as prodrugs. Methods of treating AIDS, methods of treating HIV infection, and methods of inhibiting HIV protease are described.

The compounds of the present invention are described by formula 9

where K and K 'are independently selected from H, a substituted or unsubstituted alkyl-OKd group, a cycloalkyl group, a substituted (C _g C ₆ ) alkyl group or a (C1-C ₆ ) alkyl-OH group, a heterocyclic group substituted by (C ₁ - C ₆ ) an alkyl group or (C ₁ -C ₆ ) alkyl-OH group, alkyl-KB ₂ K ₃ groups or alkyl-8 (X) (Y) K ₄ groups, where

K ₁ represents H, a substituted or unsubstituted alkyl group or an acyl group;

K ₂ and K _{3 are} independently selected from H, substituted or unsubstituted alkyl, cycloalkyl, heterocycle and aryl groups and acyl and sulfonyl groups;

K ₄ represents H, substituted or unsubstituted alkyl, cycloalkyl, heterocycle or an aryl group and each of X and Υ is absent or represents = 0;

or a pharmaceutically acceptable prodrug, salt or solvate thereof.

Preferably in the compounds of formula 9, K represents H. More preferably, K represents H and K ’represents a cycloalkyl group selected from

Preferably in the compounds of formula 9, if at least one of K and K 'is an alkyl-OKd group, K ₁ is H. In particular, when at least one of K and K' is an alkyl-0K ₁ group alkyl-OEC is selected from —C (CH3) 2CH2OH, —CH (CH3) CH2OH, —CH2CH2OH, —C (CH3) (CH2OH) 2, —C (CH3) 2 — O — CH2O — CH3, —C (CH3 ) 2-CH2-O-CH2-O-CH3 and -C (CH3) 2CH2-O-acyl, or from a pharmaceutically acceptable prodrug, salt or solvate thereof.

Preferably, if at least one of K and K 'is a cycloalkyl group, a substituted (C ₁ -C ₆ ) alkyl group or a (C ₁ -C ₆ ) alkyl-OH group, the cycloalkyl group is selected from

Preferably, if at least one of K and K 'is a heterocyclic group, a substituted (C ₁ -C ₆ ) alkyl group or a (C ₁ -C ₆ ) alkyl-OH group, the heterocyclic group is selected from

where K3 is H, substituted or unsubstituted alkyl, cycloalkyl, a heterocyclic or aryl group, or an acyl or sulfonyl group.

A preferred substance of formula 9 is [38- [2 (28 *, 38 *), 3-alpha, 4a-beta, 8abeta | | -Ν- (1,1-dimethyl-2-hydroxyethyl) -2- (2hydroxy-3 - [(3-hydroxy-2-methylbenzoyl) amino] -4- (phenylthio) butyl) -3-decahydroisoquinolinecarboxamide

or its pharmaceutically acceptable salts or similar prodrugs. Preferred prodrugs can be obtained by replacing hydrogen in one of the alcohol groups with an acyl group, and more preferably with an amino acid acyl group.

The present invention further relates to a pharmaceutical composition comprising an effective amount of a compound of formula 9 or a pharmaceutically acceptable salt thereof in combination with a pharmaceutically acceptable carrier, such as a diluent or excipient.

The present invention further relates to a method of treating AIDS, comprising administering to a recipient or patient, such as a primate, an effective amount of a compound of the present invention.

The present invention further relates to a method of inhibiting HIV replication, comprising introducing into an HIV-infected cell, a cell susceptible to HIV infection, or a recipient or patient, such as a primate, an effective amount of a compound of the present invention.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to new compounds of formula 9, as described above, which are applicable for the treatment of HIV infection and / or AIDS.

Applicants include, by reference, US patent No. 5484926, application for the grant of US patent No. 08/708411 and 08/708607 and the application for the grant of Japan patent No. W 95-248183 and W 95248184, with the proviso that the definition of preferences, terms, designations, definitions etc. used in each application are suitable only for the corresponding description from a specific application.

In particular, since each of the above applications, incorporated by reference, was compiled separately, in priority applications5 in some cases the same term, definition or variable can be used in different meanings. For example, the variable X is used in each application, but each application contains a corresponding specific value for the substituent or radical represented by this variable. It will be apparent to those skilled in the art that the terms, designations, and variables in each application incorporated by reference are limited solely to the description from the specific application and may be replaced by other suitable terms, symbols, and the like, representing certain substituents and radicals. Of course, it will be obvious to a person skilled in the art that any suitable set of terms, notation, and variables can generally or more specifically represent an object described in this appendix, including terms, notation, variables, etc., that are traditionally applied to the included descriptions in the above applications and the present description.

The compounds of formula 9 may be prodrugs that can serve to improve the pharmaceutical properties of the compounds, such as pharmacokinetic properties, for example, improve bioavailability or solubility. The manufacture of prodrugs can be carried out in accordance with standard techniques known to specialists in this field. A preferred prodrug can be obtained by acylation or alkylation of the starting alcohol, wherein I or I 'is CH (CH ₃ ) ₂ CH ₂ OH.

All temperatures indicated here are in degrees Celsius (° C). All units are indicated here in weight units, excluding liquids that are measured in units of volume.

The term alkyl, as used herein, refers to straight or branched groups, preferably containing from one to eight, more preferably containing from one to six, and most preferably containing from one to four carbon atoms. The term C ₁ -C ₆ alkyl is an unbranched or branched alkyl chain containing from one to six carbon atoms. Exemplary C ₁ -C ₆ alkyl groups include methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, neopentyl, hexyl, isohexyl, and the like. The definition of C ₁ -C ₆ alkyl includes the term C ₁ -C ₄ alkyl.

The term cycloalkyl is a saturated or partially saturated mono- or polycarbocyclic ring containing preferably 5-14 carbon atoms. Exemplary cycloalkyls include monocyclic rings containing 3-7, preferably 3-6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. A representative cycloalkyl is C ₅ -C ₇ cycloalkyl, which is a saturated hydrocarbon ring structure containing from five to seven carbon atoms.

The term alkoxyl is —O-alkyl. An example of an alkoxyl is C ₁ -C ₆ alkoxyl, which is an unbranched or branched alkyl chain containing one to six carbon atoms connected to an oxygen atom. Exemplary C1-C6 alkoxyl groups include methoxyl, ethoxyl, propoxyl, isopropoxyl, butoxyl, sec-butoxyl, tert-butoxyl, pentoxyl, hexoxyl, and the like. The definition of C ₁ -C ₆ alkoxyl includes C1-C4 alkoxyl.

The term aryl, as used herein, refers to a carbocyclic or heterocyclic, aromatic, 5-14 membered, or polycyclic ring. Exemplary aryls include phenyl, naphthyl, antryl, phenanthryl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, furyl, isothiazolyl, furazanyl, isoxazolyl, thiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl], 2- 3-b] tiantrienil, isobenzofuranyl, chromenyl, xanthenyl, fenoksatienil, indolizinyl, izoindonil, indolyl, indazolyl, purinyl, isoquinoline, quinoline, phthalazinyl, naphthyridinyl, hinoksialinil, hinzolinil, benzothiazolyl, benzimidazolyl, tetrahydroquinolinyl, cinnolinyl, pteridinyl, carbazolyl, betakarbolinil, phenan ridinil, acridinyl, perimidinyl, phenanthroline, phenazine, isothiazolyl, phenothiazines and phenoxazinyl.

The term aryloxy is —O-aryl.

The term hydrolyzable group is a group which, when substituted by the hydroxyl hydrogen of an alcohol, forms an ester that can be hydrolyzed ίη νίνο back to the corresponding alcohol. Exemplary hydrolyzable groups, which are optionally substituted, contain an acyl functional group, a sulfonate functional group, a phosphate functional group. For example, such hydrolyzable groups contain a blocked or unblocked amino acid residue, a hemisuccinate residue, and a nicotinate residue.

The term halogen represents chlorine, fluorine, bromine or iodine. The term halo- represents chloro, fluoro, bromo or iodo.

The term carbocycle is an aromatic or saturated or partially saturated 5-14 membered monocyclic or polycyclic ring, such as from a 5-7 membered monocyclic or from 7-10 membered bicyclic ring, where all members of the ring are carbon atoms.

The term heterocycle is an aromatic or saturated or partially saturated 5-14 membered monocyclic or polycyclic ring, such as a 5-7 membered monocyclic or 7-10 membered bicyclic ring containing from one to three heteroatoms selected from nitrogen atoms, oxygen and sulfur, and where any of the nitrogen and sulfur heteroatoms may optionally be oxidized and any nitrogen heteroatom may optionally be quaternized. The heterocyclic ring may be attached to any suitable heteroatom or carbon atom. Examples of such heterocycles are decahydroisoquinolinyl, octahydrothieno [3,2-c] pyridinyl, piperidinyl, piperazinyl, azepinyl, pyrrolyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazyridinyl pyrazinylpyridinyl pyrazolidinyl oxazolidinyl, isoxazolyl, thianstrenyl, triazinyl, isoxazolidinyl, morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, quinolinyl, chromenyl, xanthenyl, isoquinolazinyl, thiol benzopyranyl, benzothiazolyl, benzoazolil, furyl, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothienyl, benzo [b] thienyl, naphtho [2,3-b] thienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, oxadiazolyl, triazolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, fenoksatienil, indolizinyl, isoindolyl, indazolyl, purinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinoxyninyl, quinzolinyl, cinnolinyl, pteridinyl, carbazolyl, beta-carbolinyl, phenanthridinyl, acridinyl, perimidinyl, phenanthrolinyl, phenazinyl, phenazinyl triazinyl and phenoxazinyl.

The term thioether includes 8-aryl, such as phenylthio or naphthylthio; 8 heterocycle, where the heterocycle is saturated or partially saturated; 8- (C ₅ -C ₇ ) cycloalkyl; and 8-alkyl, such as C ₁ -C ₆ alkylthio. In thioether, -aryl, -heterocycle, -cycloalkyl and -alkyl may optionally be substituted. An example of a thioether is C ₁ C ₆ alkylthio, which is an unbranched or branched alkyl chain containing from one to six carbon atoms connected to a sulfur atom. Exemplary C1-C6 alkylthio groups include methylthio, ethylthio, propylthio, isopropylthio, butylthio, sec-butylthio, tert-butylthio, pentylthio, hexylthio and the like.

The term mercapto is -8H.

The term amino is —Pb ₁ b ₂ , where b ₁ and b _{2 are} independently selected, preferably from carbocycle, heterocycle, alkyl, sulfonyl and hydrogen; or MS (O) L ₃ , where L _{3 is} preferably alkyl, alkoxyl, hydrogen or —XL ₁ L ₂ . Aryl, alkyl and alkoxyl groups may optionally be substituted. An example of an amino is C ₁ C ₄ alkylamino, which is an unbranched or branched alkyl chain containing from one to four carbon atoms attached to an amino group. Exemplary C ₁ -C ₄ amino groups include methylamino, ethylamino, propylamino, isopropylamino, butylamino, sec-butylamino, and the like. Another example of an amino is di (C ₁ C4) alkylamino, which is two straight or branched alkyl chains, each containing from one to four carbon atoms attached to a common amino group. Exemplary di (C ₁ C4) alkylamino groups include dimethylamino, ethylmethylamino, methylpropylamino, ethylisopropylamino, butylmethylamino, sec-butylethylamino, and the like. An example of an amino is C ^ C4 alkylsulfonylamino, which has an unbranched or branched alkyl chain containing from one to four carbon atoms linked to the sulfonylamino radical. Exemplary C ₁ -C ₄ alkylsulfonylamino groups include methylsulfonylamino, ethylsulfonylamino, propylsulfonylamino, isopropylsulfonylamino, butylsulfonylamino, sec-butylsulfonylamino, tert-butylsulfonylamino, etc.

The term acyl represents T ₆ C (O) L ₄ , where L ₆ is a single bond, —O or —Ν, further, where L _{4 is} preferably alkyl, amino, hydroxyl, alkoxyl or hydrogen. Alkyl or alkoxyl groups may optionally be substituted. Exemplary acyl is C ₁ -C ₄ alkoxycarbonyl, which is a straight or branched alkoxy chain containing from one to four carbon atoms attached to a carbonyl radical. Exemplary C ₁ -C ₄ alkoxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl and the like. Another representative acyl is carboxy, where b ₆ is a single bond and b ₄ is alkoxy, hydrogen or hydroxyl. Another representative acyl is ^ (C. '| -C- |) alkylcarbamoyl (b ₆ is a single bond and b ₄ is an amino), which is a straight or branched alkyl chain containing from one to four carbon atoms attached to an atom nitrogen carbamoyl radical. Exemplary ^ (C ^ CE alkylcarbamoyl groups include Ν-methylcarbamoyl, Ν-ethylcarbamoyl, Ν propylcarbamoyl, Ν-isopropylcarbamoyl, Ν butylcarbamoyl and Ν-tert-butylcarbamoyl, etc. Another indicative acyl- _4- alkyl) , which contains two unbranched or branched alkyl chains, each of which contains from one to four carbon atoms attached to the nitrogen atom of the carbamoyl radical. Exemplary Y, M-di (C1-Sat) alkylcarbamoyl groups include Ν, Ν dimethylcarbamoyl, Ν, Ν-ethylmethylcarbamoyl, Ν, Ν-methylpropylcarbamoyl, Ν, Ν-ethylisopropylcarbamoyl, Ν, Ν-butylbutylbutylbutylamino P.

The term sulfinyl represents —§O-L ₅ , where L _{5 is} preferably alkyl, amino, aryl, cycloalkyl or heterocycle. Alkyl, aryl, cycloalkyl and heterocycle may be optionally substituted.

The term sulfonyl is —GO ₂ L ₅ , where L _{5 is} preferably alkyl, aryl, cycloalkyl, heterocycle or amino. Aryl, alkyl, cycloalkyl and heterocycle may optionally be replaced. An example of a sulfonyl is C ₁ -C ₄ alkylsulfonyl, which is a straight or branched alkyl chain containing from one to four carbon atoms attached to a sulfonyl radical. Exemplary C1-C4 alkylsulfonyl groups are methylsulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, butylsulfonyl, sec-butylsulfonyl, tert-butylsulfonyl, etc.

As indicated above, a large number of groups are optionally substituted. In fact, unless specifically indicated otherwise, all groups denoted by the terms defined in this application may or may not be substituted. For example, when the term alkyl is used, it should be understood that it covers both substituted and unsubstituted alkyl, unless one or another particular alkyl is excluded. Examples of alkyl or aryl substituents include mercapto, thioether, nitro (ΝΟ ₂ ), amino, aryloxy, halogen, hydroxyl, alkoxyl and acyl, as well as aryl, cycloalkyl and saturated and partially saturated heterocycles.

Examples of heterocycle and cycloalkyl substituents include the above substituents for alkyl and aryl, as well as aryl and alkyl.

Exemplary substituted aryls include a phenyl or naphthyl ring substituted with one or more substituents, preferably one to three substituents independently selected from halogen, hydroxy, morpholino (C ₁ -C ₄ ) alkoxycarbonyl, pyridyl (C ₁ -C ₄ ) alkoxycarbonyl, halogen (C ₁ -C ₄ ) alkyl, C ₁ -C ₄ alkyl, C ^ C ₄ alkoxy, carboxy, C-C ₄ alkoxycarbonyl, carbamoyl, No. (C '| -C' ₄ ) alkylcarbamoyl, amino, C ₁ -C ₄ alkylamino, di (C ₁ C ₄₎ alkylamino or a group of formula - (CH ₂₎ -I _and ⁷ where a is 1, 2, 3 or 4 and I ⁷ is hydroxy, C ₁ -C ₄ alkoxy, to rboksi, C ₁ -C ₄ alkoxycarbonyl, amino, carbamoyl, C ₁ -C ₄ alkylamino or di (C1-C4) alkylamino.

Another substituted alkyl is halogen (C1-C4) alkyl, which is an unbranched or branched alkyl chain containing from one to four carbon atoms with 1-3 halogen atoms attached to it. Exemplary halogen (C1-C4) alkyl groups include chloromethyl, 2-bromoethyl, 1chlorisopropyl, 3-fluoropropyl, 2,3dibromobutyl, 3-chloroisobutyl, iodine-tert-butyl, trifluoromethyl, etc.

Another substituted alkyl is hydroxy (C1-C4) alkyl, which is a straight or branched alkyl chain having from one to four carbon atoms with a hydroxy group attached to it. Exemplary hydroxy (C ₁ C4) alkyl groups include hydroxymethyl,

2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxyisopropyl, 4-hydroxybutyl and the like.

Another substituted alkyl is C ₁ -C ₄ alkylthio (C ₁ -C ₄ ) alkyl, which is a straight or branched C ₁ -C ₄ alkyl group with a C ₁ -C ₄ alkylthio group attached thereto. Exemplary C ₁ -C ₄ alkylthio (C1-C4) alkyl groups include methylthiomethyl, ethylthiomethyl, propylthiopropyl, sec-butylthiomethyl and the like.

Another representative substituted alkyl is a heterocyclic (C1-C4) alkyl, which is an unbranched or branched alkyl chain containing from one to four carbon atoms with a heterocycle attached to it. Exemplary heterocyclic (C1-C4) alkyls include pyrrolylmethyl, quinolinylmethyl, 1-indolylethyl, 2furylethyl, 3-thien-2-ylpropyl, 1-imidazolylpropyl, 4-thiazolylpropyl, and the like.

Another substituted alkyl is aryl (C1-C4) alkyl, which is an unbranched or branched alkyl chain containing from one to four carbon atoms with an attached aryl group. Exemplary arylDS ^ C 1 -alkyl groups include phenylmethyl, 2-phenylethyl, 3naphthylpropyl, 1-naphthylisopropyl, 4-phenylbutyl, etc.

The heterocycle can, for example, be substituted with 1, 2 or 3 substituents independently selected from halo, halo (C ₁ -C ₄₎ alkyl, C1C ₄ alkyl, C ₁ -C ₄ alkoxy, carboxy, C ₁ -C ₄ alkoxycarbonyl, carbamoyl , ^ U-SDalkilkarbamoila, amino, C ₁ -C ₄ alkylamino, di (C ₁ -C 4) alkylamino or a group having the structure - (CH ₂₎ -I _and ⁷ where a is 1, 2, 3 or 4 and I ⁷ represents hydroxy, C1-C4 alkoxy, carboxy, C ₁ -C ₄ alkoxycarbonyl, amino, carbamoyl, C1-C4 alkylamino or di (C1-C4) alkylamino.

Examples of substituted heterocycles include 3-Ν-tert-butylcarboxamide decahydroisoquinolinyl, ό-Ν-tert-butylcarboxamidoctahydrothieno [3,2-c] pyridyl, 3-methylimidazolyl,

3-methoxypyridyl, 4-chloroquinolinyl, 4-amino11 thiazolyl, 8-methylquinolinyl, 6-chloroquinoxalinyl, 3-ethylpropyl, 6-methoxybenzimidazolyl, 4-hydroxyfuryl, 4-methylisoquinolinyl, 6,8-dibromoquinolinyl, 2 , 3,4tetrahydroisoquinolinyl, Y-methylquinolin-2yl, 2-tert-butoxycarbonyl-1,2,3,4-isoquinolin7-yl and the like.

Exemplary heterocyclic ring systems represented by A or B include (1) 5-membered monocyclic ring groups such as thienyl, pyrrolyl, imidazolyl, pyrazolyl, furyl, isothiazolyl, furazanyl, isoxazolyl, thiazolyl and the like;

(2) 6-membered monocyclic groups such as pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, and the like;

(3) polycyclic heterocyclic ring groups such as decahydroisoquinolinyl, octahydrothieno [3,2-c] pyridinyl, benzo [b] thienyl, naphtho [2,3-b] thanthrenyl, isobenzofuranyl, chromenyl, xanthenyl and their fully or partially saturated analogues .

Cycloalkyl may optionally be substituted with 1, 2 or 3 substituents independently selected from halogen, halogen (C ₁ -C ₄ ) alkyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, carboxy, C ₁ -C ₄ alkoxycarbonyl, carbamoyl, L- (C ₁ -C ₄ ) alkylcarbamoyl, amino, C ₁ -C ₄ alkylamino, di (C ₁ C ₄ ) alkylamino or a group having the structure - (CH ₂ ) _a -B ⁷ , where a is 1, 2, 3 or 4, and B ⁷ is hydroxy, C ₁ -C ₄ alkoxy, carboxy, C ₁ C ₄ alkoxycarbonyl, amino, carbamoyl, C ₁ -C ₄ alkylamino or di (C ₁ -C ₄ ) alkylamino. Exemplary cycloalkyl groups include 3 methylcyclopentyl, 4-ethoxycyclohexyl, 5 carboxycycloheptyl, 6-chlorocyclohexyl and the like.

Exemplary hydrolyzable groups include N-benzylglycyl, L-C7-L-valyl, and N-methyl nicotinate.

The compounds of the present invention have at least five asymmetric centers, indicated by an asterisk in Formula 9 below.

As a consequence of the presence of these asymmetric centers, the compounds of the present invention can exist in any of the possible stereoisomeric forms and can be used as a mixture of stereoisomers, which can be optically active or racemic, or can be used separately as substantially pure stereoisomers, i.e. . At least 95% pure. All asymmetric forms, individual stereoisomers and combinations thereof are included in the scope of the claims of the present invention.

The individual stereoisomers can be obtained from the corresponding precursors in accordance with the methods described above, by separation of racemic mixtures or separation of diastereomers. The separation can be carried out in the presence of a separating agent, by chromatography or recrystallization, or a specific combination of these techniques known to the person skilled in the art. More details regarding the division can be found in the meeting with colleagues, Epapyoszegs, Vasetez, apb Veso Schiopz, 4o1zp APeu & 8opz 1981.

Preferably, the compounds of the present invention are substantially pure, i.e. more than 50% clean. More preferably, the compounds are at least 75% pure. Even more preferably, the compounds are more than 90% pure. Even more preferably, the compounds are at least 95% pure, more preferably at least 97% pure, and most preferably at least 99% pure.

As mentioned above, the invention relates to pharmaceutically acceptable salts of the compounds of formula 9. The compound of the present invention may contain a sufficiently acidic, sufficiently basic or both functional groups and, accordingly, interact with any of a 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 that are substantially non-toxic to living organisms. Exemplary pharmaceutically acceptable salts include salts prepared by reacting the compounds of the present invention with a mineral or organic acid or inorganic base. The reagents are usually combined in a common solvent, such as diethyl ether or benzene for the acid addition salts, or water, or alcohols for the base addition salts. Salts typically precipitate from about one hour to about ten days and can be isolated by filtration or other conventional methods. Such salts are known as acid or base addition salts.

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 ptoluenesulfonic, methanesulfonic acid, oxalic acid, p-bromophenyl sulfonic acid, carboxylic 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, caproate , oxalate, malonate, succinate, suerate, sebacate, fumarate, maleate, butyn-1,4-dioate, hexin-1,6-dioate, benzoate, chlorobenzoate, methyl benzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, , phenylpropionate, phenylbutyr t, citrate, lactate, hydroxybutyrate, glycollate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1 -sulphonates, naphthalene-2-sulfonate, mandelate and the like

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

Base addition salts include salts formed with inorganic and organic bases, such as ammonium hydroxides or alkali or alkaline earth metal hydroxides, carbonates, bicarbonates, and the like. Thus, similar bases used to prepare the salts of this invention include sodium hydroxide, potassium hydroxide, ammonium hydroxide, potassium carbonate, sodium carbonate, sodium bicarbonate, potassium bicarbonate, calcium hydroxide, calcium carbonate and the like. Potassium and sodium salts are particularly preferred.

By pharmaceutically acceptable prodrug is meant a compound that can be converted under physiological conditions or by solvolysis to a compound of formula 9.

By pharmaceutically acceptable solvate is meant a solvate that preserves the biological effectiveness and properties of the 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 should be understood that the particular counterion forming part of any salt of the present invention is not critical as long as the salt as a whole is pharmaceutically acceptable and as long as the counterion does not impart generally undesirable properties to the salt.

A preferred compound is compound 21

[38- [2 (28 *, 38 *), 3 alpha, 4a beta, 8a beta]] N- (1,1-dimethyl-2-hydroxyethyl) decahydro-2- [2hydroxy-3 - [(3-hydroxy -2-methylbenzoyl) amino] -4- (phenylthio) butyl] -3-isoquinolinecarboxamide.

A method for producing compound 21 is given below. Compound 21 was also obtained as a metabolite from the plasma of patients receiving methanesulfonate [38- (3E, 4aP *, 8aP *, 2'8 *, 3'8 *)] 2- [2'-hydroxy-3'-phenylthiomethyl- 4'-aza-5'-oxo5 '- (2' '- methyl-3-hydroxyphenyl) pentyl] decahydroisoquinoline-3-H-tert-butyl carboxamide as described in US Pat. No. 5,484,926.

The compounds of formula 9 can be obtained in accordance with the following reaction scheme I.

Reaction Scheme I

Scheme I. General pathway for the synthesis of 9b and derivatives.

2НРЗРЬ

Compound 1a, perhydroisoquinoline, which is commercially available from N80 TcbOlOg1e8 (Cysado, III) or Prgosoz 8pA (NaCl, Ala1u), undergoes prolonged acid hydrolysis in step 1a to give compound 2a. Many inorganic acids can be used either in a mixture of water / organic solvent, or only in water at temperatures above 50 ° C. An example of such an inorganic acid is 6n. aqueous HC1. Substituents of compound 1a include the corresponding esters 1b, thioesters 1c or other amides 16 ςοοζ co82

1b! With S where each of Ζ, Ζ ₁ and Ζ ₂ can independently be alkyl, cycloalkyl, heterocycle or aryl.

The amine nitrogen of compound 2a is then protected to give compound 2b in step 1b. The protecting group E _{p is} defined as a suitably joining group to avoid the undesired decomposition of the activated carboxylate derivatives of compound 2b in step 2. Such protective groups can usually be carbamate having the general structure of formula 11

About _to- och

E in Formula 11 may be any alkyl, cycloalkyl, aryl, or heterocycle that can be easily removed in the deprotection step after step 2. Examples of E include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert -butyl or higher branched or unbranched alkyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, allyl, phenyl, substituted phenyl, benzyl, substituted benzyl, 9-fluorenylmethyl, 9antrylmethyl and a higher polycyclic aromatic ring system. The following materials, as defined below, can be obtained from A1bpei Siesh1ea1 Co. (81 § A1bpey G1ika) a

I

2,2,2-trichloroethyl = - СНг ~ С-О

ABOUT

H CH

2-trimethylsilylethyl = - 0 * 2- C-CH3 n CH3 allyl = _CHg-CH = 0 * 2

Such protecting groups are usually attached via an acylation reaction of the corresponding halogen formate ester

12a or dicarbonate 12b

ABOUT

IN

WG’O X

12a, 12b

X = halogen y = OOCC 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 typically inorganic, such as metal hydroxides, bicarbonates and carbonates, or organic bases, such as amines like triethylamine, diethylamine, diethylisopropylamine, 1,8-diazabicyclo [2.2.2] octane (EABCO) or bound di- or trialkylamines, as well as amide bases like 1, 8 diazabicyclo [5.4.0] undec-7-ene (EVI) and 1.8 diazabicyclo [4.3.0] non-5-ene (ΌΒΝ). The following materials, as defined below, can be obtained from A1bpei Siesh1ea1 Co. (81 § A1bpey G1ika)

SAVCO = ^ ° θ ^υ = Οζρ “” = Of

These reactions usually occur at temperatures from below room temperature to about 100 ° C.

The amide reaction in step 2 can be carried out by any of the methods depending on how the carboxyl group is activated. Group I is added in stage 2 by the reaction of a carboxylic acid to obtain an activated derivative 2c.

Stage 2

Group I can be any of the existing many leaving groups, such as alkoxy, hydroxy, halogen, pseudohalogen (including azide, cyanide, isocyanate and isothiocyanate), alkyl or arenesulfonate, aromatic heterocycle (linked via heteroatom) and ид hydroxyheterocycle azimuth or hydroxy succin trizimide or ether. The following definitions apply to the above terms.

Ν-ΝεξΝ azide cyanide isocyanate isothiocyanate

= nitrogen-containing heterocycle alkyl sulfonate arenesulfonate

Ν-hydroxyheterocycle

M-hydroxysuccinimide hydroxybenzotriazole

Acyl halides (2c, 1 = halogen) can be prepared using inorganic halogenating agents such as thionyl chloride or bromide, trichloride or phosphorus tribromide, pentachloride or phosphorus bromide, or organic agents such as oxalyl chloride and trichloroisocyanuric acid. Esters (2c, 1 = OK) (K defined above) can be prepared in a variety of ways starting with acid chloride 2c, where I is C1, a combination requiring alcohol in the presence of an organic or inorganic base, previously added to acylate compound 12a or compound 12b. Alternatively, the ester can be obtained by esterification mediated by acid in the presence of the desired alcohol. Sulfonates (2c, 1 = O8O ₂ ^ 1, where is alkyl or aryl) are usually prepared by reacting 2b carboxylic acid with alkyl or arylsulfonyl chlorides in the presence of an organic amine base such as triethylamine in a non-polar solvent at a temperature below 0 ° C. Alkyl and arylsulfonyl are defined as follows.

O alkyl sulfoyl chloride - θ__ £ _ _{al kil} y o o

arenesulfonyl chloride = o - 3 — aryl

II about

The pseudo-halogen derivatives 2c (1 = pseudo-halogen) are usually prepared from acid halides 2c (1 = halogen) by reaction with an inorganic pseudo-halide in the presence of a base. Such bases include, but are not limited to, metal hydroxides, bicarbonates and carbonates, or organic bases such as triethylamine, diethylamine, diethylisopropylamine, 1,8-diazabicyclo [2.2.2] octane (EABCO), or related di- or trialkylamines, equal to as well as amidine bases like 1,8 diazabicyclo [5.4.0] undec-7-ene (EVI) and 1.8 diazabicyclo [4.3.0] non-5-ene (ΌΒΝ). A particularly preferred base is triethylamine. Heteroaromatic derivatives 2c are also prepared from acid halides 2c (1 = halogen) using a specific heteroaromatic compound in the presence of an amine base in a non-polar solvent. Ν-hydroxyheterocyclic derivatives of 2c can be obtained from acid halides as described above, and can also be formed using alkylcarbodiimides (alkyl # C = No alkyl, where the alkyl groups can be the same or different) or aryl carbodiimides (aryl = C = No aryl, where aryl groups may be the same or different), and amine bases as condensing agents.

The primary or secondary amine (shown under the arrow in step 2 of Scheme I) used in the reaction process may combine suitable protecting groups, depending on the functional properties determined by the amine, in the reaction method used. The type of interaction of 2c with the primary or secondary amine can be carried out in various ways, depending on the value of I. In the case where free acid is used (2c, 1 = OH), the interaction can be carried out using methods based on carbodiimides using any of the usual reagents of this class, including dicyclohexylcarbodiimide or bound dialkylcarbodiimides, EES (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide salts) or water-soluble bound reagents together with an organic amine base in polar an organic solvent such as dioxane, ΌΜΕ, ΝΜΡ, and acetonitrile in the presence of a ид hydroxyheterocyclic compound such as Ν-hydrosuccinimide or 3-hydroxybenzotriazole. Alternatively, haloformate esters, such as 12ά, can be used to temporarily activate the acid to produce mixed general anhydrides

Such haloformate esters are typical, as described above, and include methyl, ethyl, isopropyl, isobutyl, n-butyl, phenyl and associated alkyl and aryl chloroformates as defined below.

. o alkyl chloroformate =] | alkyl 0 O

O aryl chloroformate ^β aryl-O ^ O

Formula 2y is possibly intermediate in the step from formula 2b to formula 3. Formula 2ά is intermediate, but the process described here leads to formula 3 without isolating formula 2ά.

These reactions usually occur in a number of non-polar organic solvents like halogenated hydrocarbons and ethers, such as diethyl ether, methyl tert-butyl ether, diisopropyl ether, dioxane and tetrahydrofuran at temperatures below 0 ° C in the presence of an organic amine base such as triethylamine, diethylamine, diethylisophe or bound di or trialkylamines, as well as amidine bases like BVI and ΌΒΝ.

If 1 in compound 2c is an alkyl or arenesulfonate (1 = O8O ₂ or O8O ₂ Ar) interaction can occur in a number of non-polar organic solvents like halocarbons and ethers, such as diethyl ether, methyl tert-butyl ether, diisopropyl ether, dioxane and tetrahydrofuran at temperatures below 0 ° C in the presence of an organic amine base such as triethylamine, diethylamine, diethylisopropylamine, EABCO or related di- or trialkylamines, as well as amidine bases like BVI and ΌΒΝ.

If 1 in compound 2c is halogen or pseudo-halogen, the reaction can be carried out in most common organic solvents such as THF, diethyl ether, dioxane, methyl tert-butyl ether or other ethers, acetone, cyclohexanone, methyl isobutyl ketone and other ketones; esters 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, tert-butanol and related alcohols; and polar organic solvents such as dimethylformamide, dimethyl sulfoxide, Ν methyl-2-pyrrolidone and related amide-containing solvents. Typically, a base is used, which may be any of a number of inorganic bases, such as metal hydroxides, bicarbonates and carbonates, or an organic base, such as amines like triethylamine, diethylamine, diethylisopropylamine, BABCO or bound diyl trialkylamine, as well as amide bases like BVI and ΌΒΝ.

A person skilled in the art will be able to bind the amine in step 2 to the other possible 1 groups.

In step 1, deprotection can be carried out using any of the standard methods for deprotecting a particular class of protecting groups. Simple carbamates of alkyls and substituted alkyls can be removed with aqueous base solutions at temperatures up to 100 ° C using any of the simple inorganic metal hydroxides, such as sodium, lithium, potassium or barium hydroxides or other metal hydroxides in at least stoichiometric amounts. Carbamate protecting groups that contain benzyl groups bound to oxygen can be removed by hydrogenolysis on a palladium or platinum catalyst. Alternatively, basic aqueous hydrolysis can be carried out at temperatures up to about 10 ° C using any of the simple inorganic metal hydroxides, such as sodium, lithium, potassium or barium hydroxide or other metal hydroxides, at least in stoichiometric amounts. Various anhydrous acids can also be used to deprotect benzyl-based carbamates, including HC1, HBg and ΗΙ. Lewis boron and aluminum acids, such as A1C1 ₃ , BBg ₃ , BC1 _{3 are} also effective in non-polar solvents. Certain substituted benzyl, aryl or alkyl groups may also be used in which a substitution scheme is selected for their ability to be removed under specific conditions. For example, the 2-trimethylsilylethylcarbonyl group (Teos) 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 subsequently can be removed using a source of fluoride ions such as HE or tetraalkylammonium fluoride salts.

In step 4, the perhydroisoquinoline moiety of formula 4 is attached to chloro-alcohol (compound 5, Scheme I) via an intermediate epoxide (13) formed through the base-induced proximity of the hydrochloric functional center.

ZRI

Compound 5 is made Kapeka 1pbiz1psz. 1 scratch. Several closing-opening procedures can be carried out during the process compound 5 => compound 13 => compound 6. Epoxide 13 may be isolated or it may react with 4, also added subsequently, to form 13, or 4 may be from the very beginning of the process. Epoxide 13 can be prepared using inorganic bases such as metal hydroxides, carbonates and bicarbonates in solvents such as alcohols like methanol, ethanol or isopropyl alcohol, ethers such as tetrahydrofuran and dioxane, or mixtures of the two. Epoxy can also be prepared in a 2-phase solvent system consisting of water and a halocarbon solvent such as dichloromethane together with a base. A phase transfer catalyst, such as a tetraalkylammonium salt, can be used to facilitate the process. The bond breaking of the epoxide 13 with compound 4 is completed in alcoholic solvents or mixtures of alcohol and another solvent, which may be an ether or aprotic polar solvent, such as dimethylformamide or dimethyl sulfoxide. The bond between the epoxide 13 and compound 4 is broken before the release of compound 6 optimally occurs within 2–7 h at 50–60 ° С.

In step 5, the carbobenzyloxy group can be removed before the free amine 7 is released. This can be accomplished by using HBr in acetic acid using cosolvents such as halocarbons. This can also be done using boron halides, such as BBg ₃ and BC1 ₃ or alkyl substituted boron halides, such as bordemethyl bromide in halocarbon solvents like chloroform and dichloromethane at temperatures from 0 ° C to ambient temperature. Alternatively, the carboxybenzyloxy group can be removed by hydrolysis using aqueous / alcoholic solutions of metal hydroxides like barium, sodium, lithium or potassium hydroxide at temperatures above ambient temperature for several hours.

Step 6a is the reaction of benzoic acid derivatives with formula 8 to form 9a. In the formula 8, O may be a leaving group. O may be any of the leaving groups described above for group I. The compounds of formula 8, where p = OH or C1, are commercially available in EM8 OoShkop, Lep / Ligd, 8 \\ 11 / e1lps1 and 8i1a1 Ses1ca1 1pbis1pez, LM. in Japan. Linking can be given in several ways, depending on the value of p. If free acid (P = OH) is used, the coupling can be carried out using carbodiimide-based methods for the disposal of any of the usual reagents of this class, including dicyclohexylcarbodiimide or bound dialkylcarbodiimides, EOS (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide salts) or bound water-soluble reagents together with an organic amine base in polar organic solvents such as dioxane, ΌΜΕ, ΝΜΡ and acetonitrile in the presence of ид hydroxy heterocycle, including Ν-hydroxysuccinimide or 3-g drobenzotriazola. If O = a, halogen or pseudohalogen, the coupling can be carried out in most common organic solvents such as tetrahydrofuran, diethyl ether, dioxane, methyl tert-butyl ether or other ethers; acetone, cyclohexanone, methyl isobutyl ketone and other ketones; esters 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, tert-butanol and related alcohols, and polar organic solvents such as dimethylformamide, dimethyl sulfoxide, No. methyl-2-pyrrolidone and related amide-containing solvents. A base is usually used, which may be any of a number of inorganic bases, such as metal hydroxides, bicarbonates and carbonates, or an organic base, such as amines like triethylamine, diethylamine, diethylisopropylamine, OABCO or bound di- or trialkylamine, as well as amidine bases similar to ΌΒυ and ΌΒΝ.

The removal of acetate is completed in step 6b with an aqueous or alcoholic solution of inorganic bases such as metal hydroxides, carbonates and bicarbonates at temperatures from ambient temperature to 100 ° C. If the carboxamide functional group is bound to the perhydroisoquinoline ring system, then in this case it will be most easily removed (during or after stage 6b). The essence of this stage strictly depends on the type of protective group.

The preferred method for completing the entire process depicted in Scheme I is given in Scheme II. Cb / -protected amino acids are linked to amine 22 to produce amide

16. The Cb / group is removed by hydrogenation.

Scheme II

Amide Synthesis 21

OOMCVi

BUT

Stage A

OR. GA

Stage ϋ ”2 5% (M / S won and Stage C he

50% drive. Maon

Stage E

and ₃ hours tnr, then then 50% vondn. Jaon

Stage P for the preparation of amine 17. It binds to the chloroalcohol via epoxide using the ίη-zy procedure for the yield of adduct 18. Conventional deprotection of the base and coupling of the free primary amine with acid chloride 20 has led to the preparation of amide 21. This process is described in detail in Examples 1 below. A to E. The description from A to E in Scheme II is interconnected with the following Examples 1 from A to E.

The following examples illustrate aspects of the invention. These examples are used for descriptive purposes and are not intended to limit the scope of the invention.

The abbreviations for the terms: melting point, nuclear magnetic resonance spectrum, mass spectrum of electron impact, mass spectrum of field desorption, mass spectrum of bombardment by fast atoms, infrared spectrum, ultraviolet spectrum, elementary analysis, high pressure liquid chromatography and thin layer chromatography are, respectively, mp, NMR, E! M8, M8 (GE), M8 (EAB), IR, UV, electronic analysis, IRS, TLC. In addition, the absorption maximum indicated for the IR spectrum is selectable, not all maxima are indicated.

The following abbreviations are used to describe the NMR spectra: singlet (s), doublet (d), doublet of doublets (dd), triplet (t), quartet (q), multiplet (m), doublet of multiplets (dm), wide singlet (wide c) wide doublet (wide d), wide triplet (wide t), wide multiplet (wide m). I indicates the binding constant in hertz (Hz). In addition, NMR data refer to the free base of the test compound.

An NMR spectrum was obtained on a Sepeg1 E1ec1ps 9E-300 300 MHz instrument. Chemical mixing is expressed in units of δ per ppm. The mass spectrum was obtained on a US-3 spectrometer in 8 spr. Kezeag XpzbShe, Ba 1o11a, SA. The infrared spectrum was recorded on an M1bas Sogrogabop spectrometer. The UV spectrum was obtained on a Wapap Sagu 3E instrument. Thin layer chromatography was performed on silicon wafers purchased at E. Megsk. Melting points were measured on a Meil EP62 instrument and were not corrected.

Example 1. Processes for the synthesis of an amide of formula 21.

[38- [2 (28 *, 38 *), 3 alpha, 4a beta, 8a beta]] N- (1,1-dimethyl-2-hydroxymethyl) decahydro-2 [2-hydroxy-3 - [(3- hydroxy-2-methylbenzoyl) amino] -4- (phenylthio) butyl] -3-isoquinolinecarboxamide

A. Perhydroisoquinoline (26.4 g, 111 mmol) (commercially available from No. C Tesipo1o§1e§ (Cysado, 1B) or Prgozos 8pA (Miap, Pa1u)) was suspended in water (200 ml) and concentrated aqueous HC1 (200 ml ) This mixture was heated under reflux and stirred for 3 days, at which time it went into solution. Solvents were removed under reduced pressure to give a bright yellow solid. The solid was dissolved in 2-propanol (200 ml) and filtered. The filtrate was evaporated under reduced pressure to an oil. EJAc (100 ml) and water (100 ml) were added and the pH of the solution was adjusted to 8 with an additional 2N. water KOH. Benzyl chloroformate (15.8 ml, 111 mmol) was added dropwise over 30 minutes and the pH was maintained between 7 and 8 with an additional 2N. water KOH. The mixture was stirred at room temperature for 18 hours. EJAc (200 ml) was added and the organic layer was washed with 1N. aqueous HC1 (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 / ЕУАе and then 100% ЕУАс. The product containing fractions was collected and evaporated under reduced pressure to yield compound 15 (11.3 g, 32%) as a colorless oil: ¹ H NMR (300 MHz, CIS1 ₃ ) δ 7.43-7.28 (m, 5H), 5.17 (br s, 2H), 4.76 (m, 1H), 3.79 (m, 1H), 3.33 (m, 1H), 2.19 (m, 1H), 1.96 (m, 1H); 1.88-1.15 (m, 10H).

B. 1-Hydroxybenzotriazole (4.2 g, 31.4 mmol) and dichloroethane (6.0 g, 21.4 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 to 80 ° C for 10 minutes. 1,1-Dimethyl-2-trimethylsilyloxyethylamine (5.1 g, 31.4 mmol) prepared from 1,1-dimethyl-2-hydroxyethylamine (A1bpsy Sies1ca1 Co.) and hexamethyldisilazane (A1bpsy Siesh1ca1 Co.) was added by heating the mixture under reflux. for several hours, followed by evaporation of the volatile components, and the solution was heated to 80 ° C for 17 hours. The yellow solution was poured into ЕУАс (250 ml) and 2N. aqueous HC1 (250 ml). After stirring for 10 minutes, EyAc (750 ml) was added and the mixture was washed with H ₂ O (3 x 500 ml) and brine (1 x 250 ml). The resulting aqueous salts were dried (W ₂ 8 O ₄ ) and purified by thin layer chromatography (50/50 EJAe / hexane) to give compound 16 as a colorless oil (7.9 g, 78%):

¹ H NMR (300 MHz, CO 3 O) δ 7.36 (m, 5H), 5.20 (d, 1 = 8.1 Hz, 1H), 5.10 (m, 1H), 4.53 (m, 1H), 3.78 (dd, 1 = 13.2, 4.4 Hz, 1H), 3.60 (m, 2H), 3.48 (d, 1 = 10.7 Hz, 1H), 2, 15-1.25 (m, 12H), 1.31 (s, 3H), 1.29 (s, 3H).

C. A mixture of carbamate 16 (7.9 g, 20.4 mmol) and 5% palladium on carbon (Pb / C) (1.6 g) was hydrogenated at 50 psi. inch H2 in absolute CJS (110 ml) at room temperature for 18 hours. The mixture was filtered through celite and evaporated in vacuo to give amine 17 as a white crystalline solid:

¹ H NMR (300 MHz, СО3ОО) δ 3.63 (q, 1 = 7.0 Hz, 2H), 3.34 (m, 1H,) 3.27 (dd, 1 = 11.8, 3.3 Hz, 1H), 2.91 (m, 1H), 2.02-1.15 (m, 12H), 1.32 (s, 3H), 1.31 (s, 3H).

Ό. Water 10.2n. SHON (2.4 ml, 24.5 mmol) was added to a warm (27 ° C) suspension of chloroalcohol (obtained from Kapek 1pbi81peg in Japan) (10.4 g, 28.6 mmol) in isopropanol (IP) (104 ml ), mechanically mixing. After 1 h 1 n. to neutralize, aqueous HCl was added in PI (prepared by adding 1 ml of concentrated aqueous HCl to 12 ml of PI) in an amount of approximately (ex) 1 ml (pH = 7). Amine 17 (5.2 g, 20.4 mmol) was added as a solution in IP (50 ml) and a thin suspension was heated at a temperature of 60 ° С for 10 h.

IP was removed under vacuum. The residue was diluted with HEC Ac (150 ml) and washed with water (2 x 50 ml), saturated aqueous NaHCO3 (1 x 50 ml) and brine (1 x 50 ml). The combined organic layers were extracted with ESAc (1 x 25 ml). The combined organic layers were dried (No. 1 ₂ 8O ₄ ) and purified by thin layer chromatography (75/25 EJAe / hexane, then EJAc) to give compound 18 as a white solid (8.98 g, 76%):

^! H NMR (300 MHz, ΟΌ ₃ ΘΌ) δ 7.33 (m, 10H), 5.08 (AB, 1 _av = 12.2 Hz, Δυ _ΑΒ = 12.1 Hz, 2H), 3.96 (m , 2H), 3.56 (q, 1 = 7.3 Hz, 2H), 3.50 (m, 1H), 3.20 (dd, 1 = 13.6, 9.2 Hz, 1H), 3 03 (m, 1H), 2.64 (m, 2H), 2.20-1.20 (m, 14H), 1.28 (s, 6H).

E. 50% aqueous IAOH (2.7 g, 1.8 ml, 33.6 mmol) was added to a suspension of carbamate 18 (6.75 g, 11.6 mmol) in PI (34 ml) at room temperature. The mixture was refluxed for 12 hours. After cooling to room temperature, the mixture was diluted with methyl tert-butyl ether (MTBE) (600 ml) and washed with H2O (2 x 250 ml) and brine (1 x 125 ml). The combined organic layers were extracted with MTBE (1 x 150 ml). The combined organic layers were dried (Na ₂ 8 O ₄ ) and evaporated in vacuo to give a mixture of compound 19 and benzyl alcohol as an oily white solid.

¹ H NMR (300 MHz, C1) _; O1)) δ 7.34 (m, 10H), 4.63 (s, 2H), 3.81 (m, 1H), 3.58 (m, 3H), 3.03-2.60 (m, 5H), 2.17 (m, 1H), 2.05 (m, 1H), 1.87-1.05 (m, 12H), 1.30 (s, 3H), 1.28 (s, 3H )

E. Triethylamine (3.2 g, 4.3 ml, 31.2 mmol) is added to a solution of a mixture of amine 19 (4.7 g, 10.4 mmol theoretically from 18) and benzyl alcohol in ENS (23 ml) at room temperature. Then a solution of 3-acetoxy-2methylbenzoyl chloride (20) was added (obtained according to the processes described in the appendix to US Pat. No. 8,171, No. 08 / 708,411, published September 5, 1996, here specially included in the appendix) (2.4 g, 11.5 mmol) in tetrahydrofuran (4 ml). After 2 hours, 50% aqueous NO was added (4.1 g, 2.8 ml, 52.2 mmol) and the mixture was refluxed for 1 hour. After cooling to ambient temperature, the pH of the mixture was neutralized to 7 at help 2n. aqueous HC1 (26 ml). This mixture was diluted with EyAc (500 ml) and washed with H ₂ O (1 x 250 ml), saturated aqueous HCO ₃ (2 x 250 ml), H ₂ O (1 x 250 ml) and brine (1 x 125 ml). The organic layer was dried (Na ₂ 8 O ₄ ) and purified by thin layer chromatography (75/25 EJAc / Hexane) to give amide 21 as a white foam (1173-57A, 1.39 g, 23%).

¹ H NMR showed the presence of 11 wt.% EJAc, which was impossible to remove under vacuum.

Analysis:

¹ H NMR (300 MHz, C1EO1)) δ 7.53 (d, 1 = 7.3 Hz, 2H), 7.32 (t, 1 = 7.0 Hz, 2H), 7.20 (t, 1 = 7.3 Hz, 1H), 7.06 (t, 1 = 8.1 Hz, 1H), 6.92 (d, 1 = 8.1 Hz, 1H), 6.83 (d, 1 = 8 , 1 Hz, 1H), 4.42 (m, 1H), 4.08 (m, 1H), 3.61 (dd, 1 = 13.6, 4.0 Hz, 1H), 3.45 (AB , 1 _AB = 11.0 Hz, Δυ ·, 18.0 Hz, 2H), 3.29 (dd, 1 = 13.6, 10.3 Hz, 1H), 3.10 (m, 1H), 2 66 (m, 2H), 2.28 (s, 3H), 2.22 (m, 2H), 2.04 (m, 1H), 1.86-1.20 (m, 11H), 1, 19 (s, 3H), 1.18 (s, 3H).

¹³ C NMR (75.5 MHz, C1EO1)) δ 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 activity and anti-HIV activity in the cell culture of compound 21.

To determine the Κι value in compound 21, a method for analyzing the kinetics of strong binding was used. Κι = 5.6 ± 0.91 nM.

Ways

Expression of HIV-1 protease.

The HIV-1 protease gene was isolated from the ΙΙΙΒ viral extract (Ka1pet, L., et al., Iaige, 316, 227-284 (1985)). In order to enhance the stability of the purified protease (Koke, 1.K. et al., 1. Vu1. Sies., 268, 11939-11945 (1993)), the glutamine residue at position 7 (07) was replaced by serine (8) by The 33 main pairs of segments between the II1 sites and BK1EP sites of the protease gene sequence with synthetic oligonucleotides encoded by mutation (078). The modified gene sequence was introduced into the plasmid vector ρΟΖ (Mepde, K.L. et al., Vyusyeshshyu, 34: 15934-15942 (1995)) under the control of the phage of the T7 promoter. The resulting construct, ρ6Ζ / 4Ρ-19Ο78 # 9, was introduced into E. soybean B21 strain (IE3), acquired in Iowade, 1ps.

Expression of HIV-1 RK: Cultures were grown in 2 1,6 medium (1.6% Turisac Peplop, 1% Geaca ex1tac1, 0.5% IaC1 with an initial pH of 7.5) containing 200 μg / L ampicillin in a 100 L fermenter (Vu1aE111e 8A ) at 37 ° C for 5 hours and then induced by the addition of 1 mM ISTG (isopropyl-β-ти-thiogalactopyranoside). The culture temperature during induction was increased to 42 ° C to increase the accumulation of recombinant HIV-1 protease as insoluble inclusions. After 2 hours at 42 ° C, the cells were collected by cross-filtration using a 0.1 μm Reshcop UURP000C5 cassette # 10 (M1 Schroge), and the cell paste was frozen and left at -70 ° C.

Purification of Recombinant Protease

HIV-1: All stages, unless specifically indicated, were carried out at 4 ° C.

Protein concentrations were measured using VUKai protein analysis solution with bovine serum albumin (VU27

Nab, Βίο1ιιηοπ6. CA) as a standard. The stages of chromatography and the purity of HIV pH were measured by dodecyl sulfate sodium polyacrylamide gel electrophoresis (8–8 –6–). The final purity of HIV pH was> 98%. The average final yield for every 100 L of culture was ~ 120 mg.

Cell paste from 100 L of culture was resuspended in 300 ml of lysis buffer (50 mM Tris-C1 pH 8.0. 25 mM IaC1, 20 mM 2 mercaptoethanol) and microfluidized in M | sgoPshb5 Sotrotab fluidizer at 22,000 psi. inch. The crude cell lysate was clarified by centrifugation at 14,000 rpm for 20 minutes. In the sediment after centrifugation, the majority were inclusions of HIV pH. The inclusions were sequentially washed several times in a lysis buffer containing an additional 0.1 Ττίΐίοη-Χ100 and 1 M urea and, after the washing process. inclusions were precipitated by centrifugation at 5000 rpm for 20 minutes The purified inclusions were dissolved in a buffer containing 50 mM Tris-C1, pH 8.0. 25 mM IA1. 20 mM 2-mercaptoethanol and 8 M urea. The solution was clarified by centrifugation at 14,000 rpm and, at room temperature, it was poured into a 300 ml high-speed column on ₉ Sepharose (Ryatshasha. Rksa1a \ yau. N1). balanced with the same buffer. Under such conditions, HIV pH does not bind to the column and the practically purified enzyme is in the flow fraction. To renature protein, fractions from a fast flow column on O-Sepharose were dialyzed against three exchangeable buffers containing 25 mM NaΗ ₂ Η ₄ pH 7.0. 25 mM No. C1. 10 mM DDT and 10% glycerol. After distillation, small amounts of precipitate were removed by centrifugation, and the obtained enzyme preparation was concentrated and dialyzed against 0.5 No. C1. 50 mM ΜΕ8 pH 5.6. 10 mM DDT were frozen in small aliquots of ~ 2 mg / ml and left at -70 ° C.

Evaluation of the kinetics of strong binding and its analysis.

The proteolytic activity of purified HIV-1 protease was measured by a modified chromogenic assay developed by H1syatb8 et al. (Ksjatbk. Α.Ό. et al., 1. Βίο1. Here we have. 256. 773-7736 (1990)). Synthetic protein H18-Eu8-A1a-Arg-Ua1-EEI-Rye (PA1 ^ O ₂₎ S1i-A1a-Me-8eg ^ _H2O (Rerbbe Sotrapu americana) (Me - norleucine) was used as substrate. The analysis was carried out in 0.5 M No. C1. 50 mM ΜΕ8 pH 5.6. 5 mM DDT, and 2% DMSO at 37 ° C. The cleavage of the cleaved bond between leucine and paranitrophenylalanine (RNe rapaΝΟ ₂ ) was analyzed by spectrophotometric monitoring of the decrease in absorption at 305 nm. The initial rate was determined by the level of decrease in absorption during the first 100 s of the enzymatic reaction. In these conditions and when using the 078 HIV-1 protease, the Michaelislis constant (KM) for this substrate was 59 ± 17 μm.

To stop the inhibition of compound 21, saturation of the substrate concentration at 200 μm was applied. Between 13 and 20, inhibitor concentrations were measured, and the reaction rate was measured for each concentration, as described above. Found Κί (Κί found.). the aforementioned was determined by computer nonlinear fitting in small areas by the method of tightly coupled Morrison alignment (Μοττίκοη. ЕЕ .. VyusNesh. Vyur. Ac1a. 185. 269-286 (1963)).

Example 3. Antiviral activity of compound 21 against HIV-1 in cell culture.

Cellular and viral strains.

The lines of CEM-88 and MT-2 of human T-cells and strains of HIV-1 HE and ΙΙΙΒ are obtained from AJ8 Neckeksn apb Negegeps Rogodgash. Όίνίδίοη GO AGO8. MATO and MN.

Cell defense assays.

The inhibitory effects of each agent on HIV-1 replication were measured by a method of reducing MTT mortality (A11eu. M.S. et al., Sapseg Nek. 48: 589-601 (1988)). The compounds were dissolved in DMSO at a concentration of 40 mg / ml, then diluted with a 1: 200 medium culture (HPM1. Supplemented with 10% germinal bovine serum). 100 μl was taken from each stock diluted product and transferred to a 96 well plate where standard double dilutions were made. In various tubes, MT-2 cells and CEM-88 cells infected with HIV1 ΙΙΙΒ or HIV-1 NOT with multiple infections (MH) at 0.01 and 0.03, respectively. After a 4-hour adsorption period, 100 μl of infected or uninfected cells was added to the wells of the plate containing the drug to obtain a final concentration of 1 x 10 ⁴ cells / well. After 6 (CEM-88 cells) or 7 (MT-2 cells) days, MTT (5 mg / ml) was added to the test boards, and the resulting formazan was measured spectrophotometrically at 570 nm. Data were expressed as a percentage of those formed in non-contaminated drug-free wells, formazan. ΕΌ ₅₀ was calculated when the amount of formazan formed in infected, drug-treated cells reached 50% of that formed in uninfected, non-drug-treated cells. The therapeutic index (TI) was calculated by dividing cytotoxicity (CT ₅₀ ) by antiviral efficacy (ΕΌ ₅₀ ).

Table 1

Antiviral activity and cytotoxicity estimates of compound 21 in acute infection of CEM-88 HIV-1 KE cells

Compound ΕΏ50, nm EE. 5, nm ^CT 50 ^, microns The therapeutic index ^and 21 34.2 154.1 96.6 2825 Azidothymidine (AZT) 52.3 543.1 > 374.5 > 7161 Dideoxycytidine (DDS) 94.70 142.0 37.69 398

^a Therapeutic index = Cytotoxicity (CT ₅₀ ) Antiviral activity (EO ₅₀ ).

table 2

Antiviral activity and cytotoxicity estimates of compound 21 in acute infection of HIV-1 MT-2 cells ΙΙΙΒ

Compound E1) x „ nm ΕΏ95, nm μm The therapeutic index ^and 21 85.6 ΝΏ 92.6 1082 AZT 430.7 ΝΏ 109,4 254 DDS 5924 ΝΏ 176.3 thirty

^a Therapeutic index = Cytotoxicity (CT ₅₀ ) Antiviral activity (Εϋ ₅₀ ).

As indicated above, the compounds of the present invention are used to inhibit HIV protease, which is an enzyme related to the production and assembly of viral components. An embodiment of the present invention is a method of treating HIV infection, comprising administering to a carrier or patient, such as a primate, an effective amount of a compound of formula 9 or a pharmaceutically acceptable salt thereof. Another embodiment of the present invention is a method of treating AIDS, comprising administering to a carrier or patient, such as a primate, an effective amount of a compound of formula 9 or a pharmaceutically acceptable salt thereof. A further embodiment of the present invention is a method for inhibiting an HIV protease, comprising administering to a carrier 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 a pharmaceutically acceptable salt thereof, effective to inhibit the production and assembly of viral components associated with HIV protease. The specific dose of the compound administered according to the present invention gives a therapeutic or inhibitory effect will, of course, be determined by various circumstances defined by each situation, including, for example, the compound administered, the route of administration, the condition in which the carrier or patient is located, or its individual characteristics . An exemplary daily dose (administered simultaneously or divided) contains a dosage level of from about 0.01 mg / kg to about 50 mg / kg body weight of the compound of the present invention.

Preferred daily doses are generally from about 0.05 mg / kg to about 40 mg / kg, and more preferably from about 1.0 mg / kg to about 30 mg / kg.

The compounds of the invention can be administered in a variety of ways, including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular and intranasal methods. The compounds of the present invention are preferably formed prior to administration. Moreover, another embodiment of the present invention is a pharmaceutical composition 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 excipient thereof.

The active ingredient preferably contains from 0.1 to 99.9% by weight of the composition. By pharmaceutically acceptable is meant that the carrier, such as a diluent or excipient, is compatible with the rest of the ingredients of the mixture and is not toxic to the carrier or patient.

Pharmaceutical mixtures can be prepared from the compounds of the invention by known procedures using known and widely available ingredients. In preparing the compositions of the present invention, the active ingredient is typically mixed with a carrier or dissolved in a carrier, or enclosed in a carrier, which may be in the form of a capsule, sachet, paper or other suitable container. When the carrier acts as a solvent, it can be a solid, semi-solid, or liquid material that acts as a carrier, excipient, or medium for the active ingredient. Thus, the mixture can be in the form of tablets, pills, powders, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (both in solid and in liquid medium), ointments (containing, for example, up to 10% by weight of active compounds), soft or hard gelatin capsules, suppositories, sterile injectable solutions, sterile powders, etc.

The following composition examples are illustrative only and are not intended to limit the scope of the invention. The term active ingredient is a compound of formula 9 or a pharmaceutically acceptable salt thereof.

Composition 1.

Hard gelatin capsules are prepared using the following ingredients:

Amount, mg / capsule Active ingredient 250 Dry starch 200 Magnesium stearate 10 Total 460 mg

Composition 2.

A tablet is prepared using the following ingredients:

Amount, mg / tablet Active ingredient 250 Monocrystalline cellulose 400 Smoked silica 10 Stearic acid 5 Total 665 mg

Composition 5.

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

Amount, mg / capsule Active ingredient 80 mg Starch 59 mg Monocrystalline cellulose 59 mg Magnesium stearate 2 mg Total 200 mg

The components are mixed and pressed into tablets, each weighing 665 mg.

Composition 3.

Aerosol solution is prepared from the following components:

Weight Active ingredient 0.25 Methanol 25.75 Propellant 22 (chlorodifluoromethane) 74.00 Total 100.00

The active components are mixed with ethanol and the mixture is added to a portion of poropellant 22, cooled to -30 ° C and transferred to a filling apparatus. Then, the required amount is introduced into a stainless steel container and diluted with propellant residues. The valve apparatus is attached to the container later.

Composition 4.

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

Amount, mg / tablet Active ingredient 60 Starch 45 Monocrystalline cellulose 35 Polyvinylpyrrolidone (as a 10% solution in water) 4 Carboxymethyl Sodium Starch 4,5 Magnesium stearate 0.5 Talc one Total 150

The active ingredient, starch and cellulose are passed through Νο. 45 nIS and 8. strainer and mix thoroughly. An aqueous solution containing polyvinylpyrrolidone is mixed with the obtained powder and the mixture is passed through Νο. 45 nIS and 8. sieve. The granules thus obtained are dried at 50 ° C. and passed through Νο. 18 shekels and 8. sieve. Carboxymethyl sodium starch, magnesium stearate and talc are previously passed through Νο. 60 shekels and 8. a sieve and then added to the granules, which, after mixing, are pressed in a tablet machine to produce tablets each weighing 150 mg.

The active ingredient, cellulose, starch and magnesium stearate are mixed, passed through Νο. 45 nIS and 8. sieve and placed in hard gelatin capsules in an amount of 200 mg.

Composition 6.

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

Active ingredient 225 mg Saturated Fatty Acid Glycerides 2000 mg Total 2225 mg

The active ingredient is passed through Νο. 60 shekels and 8. sieve and suspended in saturated fatty glycerides, pre-melting them using the minimum amount of heat. The mixture is then poured into a 2 g suppository mold and cooled.

Composition 7.

Suspensions containing each 50 mg of the active ingredient, 5 mg each are prepared as follows:

Active ingredient 50 mg Sodium carboxymethyl cellulose 50 mg Syrup 1.25 mg Benzoic acid solution 0.10 mg Sweetener number of dec. Dye number of dec. Pure water to the total volume 5 ml

The active ingredient is passed through Νο. 45 nIS and 8. sieve and mixed with sodium carboxymethyl cellulose and syrup until a uniform paste is formed. The benzoic acid solution, sweetener and dye are dissolved in a portion of water and added by stirring. The necessary water is added until the desired volume is obtained.

Composition 8.

The composition for intravenous administration is prepared as follows:

Active ingredient 100 mg Isotonic saline 1000 ml

A solution of the above ingredients is typically administered to a patient at a rate of 1 ml / min.

Composition 9.

A tablet is prepared using the following ingredients:

Amount, mg / tablet Active ingredient 292 mg Calcium silicate 146 mg Crospovidone 146 mg Magnesium stearate 5 mg Total 589 mg

CLAIM

Claims (10)

CLAIM 1. The compound of formula n or a pharmaceutically acceptable salt or solvate thereof. 2. The compound according to claim 1, having a purity of more than 90%. 3. The compound according to claim 1, having a purity of at least 95%. 4. The compound according to claim 1, having a purity of at least 97%. 5. The compound according to claim 1, having a purity of at least 99%. 6. A pharmaceutical composition for treating HIV infection comprising an effective amount of a compound or a salt or solvate thereof according to claim 1 and a pharmaceutically acceptable carrier. 7. The pharmaceutical composition according to claim 6, where the compound or its salt or MES have a purity of more than 90%. 8. The pharmaceutical composition according to claim 6, where the compound or its salt or MES have a purity of at least 95%. 9. The pharmaceutical composition according to claim 6, where the compound or its salt or MES have a purity of at least 97%. 10. The pharmaceutical composition according to claim 6, where the compound or its salt or solvate have a purity of at least 99%.