3-{(3R, 4R)-4-Methyl-3-[methyl-(7h-pyrrolo [2,3-d] pyrimidin-4-yl )amino] piperidin-1-yl) }-3-oxo propionitrile, pharmaceutical composition containing thereof and its use

3 - {(3R, 4R) -4-Methyl-3- [methyl- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino] piperidin-1-yl}} - 3-oxopropionitrile, pharmaceutical composition its content and use

Field of the invention

The present invention relates to pyrrolo [2,3-d] pyrimidine compounds, pharmaceutical compositions containing them and the use of these compounds as protein kinase inhibitors, such as the Janus Kinase enzyme (Janus Kinase 3 = JAK3).

BACKGROUND OF THE INVENTION

The pyrrolo [2,3-d] pyrimidine compounds are inhibitors of protein kinases such as the Janus kinase 3 (JAK.3) enzyme and are therefore useful as immunosuppressive agents for organ transplantation, xenotransplantation, in the treatment or prevention of a condition selected from the group consisting of lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, type I diabetes and complications of diabetes, cancer, asthma, atopic dermatitis, autoimmune thyroid disorders, ulcerative colitis, Crohn's disease, Alzheimer's disease, leukemia, and other indications in which immunosuppression is desired. These pyrrolo [2,3-d] pyrimidine compounds, pharmaceutical compositions containing such compounds and methods of using the compounds are described in co-pending patent application Ser. No. 09/732669, filed Dec. 8, 2000, owned by the same proprietors as the present invention. . Initially, racemic mixtures of these pyrrolo [2,3-d] pyrimidine compounds are obtained, with the individual enantiomers in substantially isolated pure form being preferred for use as medicaments, and in some cases such forms are necessary for use as medicaments. By using a specific compound precursor in the synthesis of these compounds, the stereochemistry of the desired compounds can be predetermined. The methods of the present disclosure relate to substantial chiral resolution of the salts of racemic mixtures of precursor compounds used to prepare the individual enantiomeric forms of these pyrrolo [2,3d] pyrimidine compounds.

SUMMARY OF THE INVENTION

The scope of the invention is defined in the claims. In particular, the present invention provides a compound which is 3 - {(3R, 4R) -4-methyl-3- [methyl- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino] piperidin-1- yl) -3} oxopropionitrile or a pharmaceutically acceptable salt thereof.

The present invention also provides a pharmaceutical composition comprising a proportion of the above compound or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable carrier.

According to a preferred embodiment of the present invention there is provided a pharmaceutical composition comprising a proportion of the above-mentioned compound or a pharmaceutically acceptable salt thereof in combination with one or more other agents that modulate the mammalian immune system or anti-inflammatory agents and a pharmaceutically acceptable carrier.

Finally, the present invention provides the use of the above compound or a pharmaceutically acceptable salt thereof, alone or in combination with one or more other agents that modulate the mammalian immune system or anti-inflammatory agents in an effective amount for the manufacture of a medicament for inhibiting protein kinases or Janus kinase (JAK3). in a mammal, including a human.

A preferred embodiment of the present invention is the use of the above compound or a pharmaceutically acceptable salt thereof, alone or in combination with one or more additional agents that modulate the immune system of a mammal or an anti-inflammatory agent in an effective amount for manufacture

Drugs for treating or preventing a disorder or condition selected from the group consisting of organ transplant rejection, xenotransplantation, lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, type I diabetes and diabetes complications, cancer, asthma, atopic dermatitis , autoimmune thyroid disorders, ulcerative colitis, Crohn's disease, Alzheimer's disease, leukemia, and other autoimmune diseases occurring in a mammal, including humans.

The following is a more detailed description of the present invention, comparative data and other information intended to illustrate the full scope of the solution upon which the present invention is based.

The present invention is made possible by a process for the resolution of the enantiomers of the precursor used to prepare the compounds of the following general formula, and in particular to form the R group¹ in this compound:

<img file="CZ304366B6_D0001.tif" />

<img file="CZ304366B6_D0002.tif" />

or a pharmaceutically acceptable salt thereof, wherein in the above general formula: R¹ means a group of the general formula:

<img file="CZ304366B6_D0003.tif" />

in which:

y is 0, 1 or 2,

R⁴ is selected from the group consisting of hydrogen, alkyl of 1 to 6 carbon atoms, alkylsulphonyl of 1 to 6 carbon atoms, alkenyl of 2 to 6 carbon atoms, alkynyl of 2 to 6 carbon atoms, the alkyl group, alkenyl the alkynyl group may be optionally substituted with deuterium, hydroxy, amino, trifluoromethyl, (C 1 -C 4) alkoxy, (C1 -C6) acyloxy, (C1 -C6) alkylamino, (alkyl)₂an amino group having 1 to 6 carbon atoms in each alkyl moiety, a cyano group, a nitro group, an alkenyl group having 2 to 6 carbon atoms, an alkynyl group having 2 to 6 carbon atoms, or an acylamino group having 1 to 6 carbon atoms, or R⁴ represents a cycloalkyl group having 3 to 10 carbon atoms, said cycloalkyl group being optionally substituted by deuterium, hydroxy, amino, trifluoromethyl, alkoxy having 1 to 6 carbon atoms, acylamino having 1 to 6 carbon atoms, alkylamino having 1 to 6 carbon atoms carbon atoms, (an alkyl-amino group having from 1 to 6 carbon atoms in each alkyl moiety, cyano, cyanoalkyl having 1 to 6 carbon atoms in the alkyl moiety, trifluoromethylalkyl having 1 to 6 carbon atoms in the alkyl moiety, nitro, nitroalkyl

-2CZ 304366 B6 having 1 to 6 carbon atoms in the alkyl moiety or 1 to 6 carbon acylamino,

R⁵ means a heterocycloalkyl group having 1 to 9 carbon atoms, the heterocycloalkyl groups having to be substituted by one to five carboxy, cyano, amino, deuterium, hydroxy, alkyl groups having 1 to 6 carbon atoms, alkoxy groups having 1 to 6 carbon atoms, halogens, (C 1 -C 6) acyl groups, (C 1 -C 6) alkylamino groups, (C 1 -C 6) aminoalkyl, (C 1 -C 6) alkoxy-CO-NH (C 1 -C 6) alkoxy, (C 1 -C 6) alkylamino-CO-, (C 2 -C 6) alkenyl, (C 2 -C 6) alkynyl, (C 1 -C 6) alkylamino, (C 1 -C 6) aminoalkyl, (C1-C6) hydroxyalkyl, (C1-C6) alkoxyalkyl and (C1-C6) alkyl, (C1-C6) acyloxyalkyl and (C1-C6) alkyl moieties, nitro groups, cyanoalkyl groups having 1 to 6 carbon atoms, haloalkyl groups having 1 to 6 carbon atoms, nitroalkyl groups containing 1 to 6 carbon atoms in the alkyl moiety, trifluoromethyl groups, trifluoromethyl alkyl groups having 1 to 6 carbon atoms in the alkyl moiety, acylamino groups having 1 to 6 carbon atoms, acylaminoalkyl groups containing 1 to 6 carbon atoms in the alkoxy moiety and 1 to 6 carbon atoms 6 carbon atoms in the alkyl moiety, alkoxyacylamino groups having 1 to 6 carbon atoms in the alkoxy moiety and 1 to 6 carbon atoms in the acyl moiety, (C 1 -C 6) aminoacyl, (C 1 -C 6) aminoacylalkyl (C 1 -C 6) -alkyl, (C 1 -C 6) alkylaminoacyl (C 1 -C 6) alkyl (Aminoacyl) groups containing 1 to 6 carbon atoms in each alkyl moiety and 1 to 6 carbon atoms in the acyl moiety, R groups¹⁵R^I6N-CO-O-;^,5R^I6N-CO (C1-C6) alkyl groups, alkyl-S (O) m (C1-C6) alkyl groups, R groups¹⁵R¹⁶NS (O) m, R.¹⁵R¹⁶NS (O) alkyl groups containing 1 to 6 carbon atoms in the alkyl moiety;¹⁵S (0) mR¹⁶N, R¹⁵S (O)_mR¹⁶N-alkyl groups containing up to 6 carbon atoms in the alkyl moiety, wherein m is 0, 1 or 2;

R¹⁵ and R¹⁶ each independently selected from the group consisting of hydrogen and alkyl having 1 to 6 carbon atoms, or a group of formula II:

<img file="CZ304366B6_D0004.tif" />

wherein a is 0, 1, 2, 3 or 4, b, c, e, f and g are each independently 0 or 1,

D is 0, 1, 2 or 3,

X is S (O)_nin which n is 0, 1 or 2, an oxygen atom, a carbonyl group or a -C (= N-cyano) - group,

Y is S (O)_nwherein n is 0, 1 or 2, or a carbonyl group, and

Z is carbonyl, C (O) O-, C (O) NR- or S (O)_nwherein n is 0, 1 or 2,

R⁶, R⁷, R⁸, R⁹, R¹⁰ and R¹¹ are independently selected from the group consisting of hydrogen and alkyl of 1 to 6 carbon atoms, optionally substituted with deuterium, hydroxy, amino, trifluoromethyl, acyloxy of 1 to 6 carbon atoms, acylamino of 1 to 6 carbon atoms, alkylamino (C 1 -C 6) alkyl group (C 1 -C 6 alkyl-amino group in each alkyl moiety, cyano, (C 1 -C 6) cyanoalkyl, (C 1 -C 6) trifluoromethyl alkyl, nitro, (C 1 -C 6) nitroalkyl and (C 1 -C 6) acylamino,

R¹² means carboxy, cyano, amino, oxo, deuterium, hydroxy, trifluoromethyl, C 1 -C 6 alkyl, C 1 -C 6 trifluoromethyl, C 1 -C 6 alkoxy, halo, acyl (C 1 -C 6) group, (C 1 -C 6) alkylamino group, (alkyl) a 2-amino group having 1 to 6 carbon atoms in each alkyl moiety, an aminoalkyl group having 1 to 6 carbon atoms, an alkoxy-CO-NH group having 1 to 6 carbon atoms in the alkoxy moiety, an alkylaminoCO- group having 1 to 6 atoms carbon in the alkyl moiety, alkenyl of 2 to 6 carbon atoms, alkynyl of 2 to 6 carbon atoms, alkylamino of 1 to 6 carbon atoms, hydroxyalkyl of 1 to 6 carbon atoms, (C 1 -C 6) alkoxyalkyl, both C 1 -C 6 acyloxyalkyl, C 1 -C 6 acyloxyalkyl, nitro, C 1 -C 6 cyanoalkyl, haloalkyl (C 1 -C 6) group, (C 1 -C 6) nitroalkyl, (trifluoromethyl), (C 1 -C 6) trifluoromethyl (C 1 -C 6) alkyl group, (C 1 -C 6) acylamino, (C 1 -C 6) acylaminoalkyl, both in the acyl moiety and in the alkyl moiety, (C 1 -C 6) alkoxyacylamino moiety in both the alkoxy and acyl moieties; (C-C uhlíku), (C až-C amino amino) aminoacylalkyl, C až-C alky alky (Clamino-C alky alky) alkylaminoacyl group, (alkylEaminoacyl having 1 to 6 carbon atoms in each alkyl moiety and 1 to 6 carbon atoms in the acyl moiety, R^I5R^I6N-CO-OR¹⁵R¹⁶An N-CO-alkyl group having from 1 to 6 carbon atoms in the alkyl moiety;¹⁵C (O) NH;¹⁵OC (O) NH, R^I5NHC (O) NH, alkyl-S (O) m containing 1 to 6 carbon atoms in the alkyl moiety, alkyl-S (O)_m-alkyl containing 1 to 6 carbon atoms in each alkyl moiety;^l5R¹⁶NS (O) m, R.^l5R^l6NS (O) m-alkyl containing 1 to 6 carbon atoms in the alkyl moiety, R^l5S (0) mR¹⁶N, R^l5S (0) mR¹⁶N-alkyl having 1 to 6 carbon atoms in the alkyl portion wherein m is 0, 1 or 2, and R is¹⁵ and R¹⁶ are each independently selected from the group consisting of hydrogen and alkyl of 1 to 6 carbon atoms,

R² and R³ each independently selected from the group consisting of hydrogen, deuterium, amino, halogen, hydroxy, nitro, carboxy, alkenyl of 2 to 6 carbon atoms, alkynyl of 2 to 6 carbon atoms, trifluoromethyl, trifluoromethoxy, alkyl a group containing from 1 to 6

C 1-4 alkoxy, C 1 -C 6 alkoxy, C 3 -C 10 cycloalkyl, wherein the alkyl, alkoxy or cycloalkyl group may be optionally substituted with one to three substituents selected from the group consisting of halogen, hydroxy, carboxy, (C 1 -C 5) aminoalkylthio, (C 1 -C 8) alkylamino, (alkyl)₂an amino group having 1 to 6 carbon atoms in each alkyl moiety, a heteroaryl group having 5 to 9 carbon atoms, a heterocycloalkyl group having 2 to 9 carbon atoms, a cycloalkyl group having 3 to 9 carbon atoms, or an aryl group having 6 to 10 carbon atoms, or R² and R³ each independently represent a C 3 -C 10 cycloalkyl, C 3 -C 10 cycloalkoxy, C 1 -C 6 alkylamino, (C 1 -C 6 alkylamino), arylamino (C 6 -C 10) group, (C 1 -C 10) alkylthio, (C 6 -C 10) arylthio group, (C 1 -C 6) alkylsulfinyl, (C 6 -C 10) arylsulfinyl, (C 1 -C 6) alkylsulfonyl, (C 6 -C 10) arylsulfonyl, (C 1 -C 6) acyl, alkoxy-CO-NH - a group having 1 to 6 carbon atoms in the alkoxy moiety, an alkylamino-CO- group having 1 to 6 carbon atoms in the alkyl moiety, a heteroaryl group having 5 to 9 carbon atoms, C 2 -C 9 heterocycloalkyl or C 6 -C 10 aryl, wherein the heteroaryl, heterocycloalkyl and aryl groups are optionally substituted with one to three substituents selected from the group consisting of halogen, C 1 -C 6 alkyl, alkyl A -CO-NH group having 1 to 6 carbon atoms in the alkyl moiety, an alkoxy-CO-NH- group having 1 to 6 carbon atoms in the alkoxy moiety, alkyl-CO-NH-alkyl having 1 to 6 carbon atoms in each alkyl moiety, alkoxy-CO-NH-alkyl having 1 to 6 carbon atoms in both the alkoxy moiety and the alkyl moiety, alkoxy-CO-NH- an alkoxy group having 1 to 6 carbon atoms in each alkoxy moiety, a carboxy group, a carboxyalkyl group having 1 to 6 carbon atoms in the alkyl moiety, a carboxyalkoxy group having 1 to 6 carbon atoms in the alkoxy moiety, benzyloxycarbonylalkoxy having 1 to 6 carbon atoms in the alkoxy moiety, alkoxycarbonylalkoxy having 1 to 6 carbon atoms in each alkoxy moiety, aryl having 6 to 10 carbon atoms, amino, aminoalkyl having 1 to 6 carbon atoms in the alkyl moiety, alkoxycarbonylamino (C 1 -C 6) alkoxy, (C 1 -C 6) alkoxycarbonylamino (C 6 -C 10) aryl group; (C 1 -C 6) alkylamino (C 1 -C 6 alkylamino), C 1 -C 6 alkylaminoalkyl, (alkyl)₂(C 1 -C 6) aminoalkyl, hydroxy, (C 1 -C 6) alkoxy, carboxy, (C 1 -C 6) carboxyalkyl, (C 1 -C 6) alkoxycarbonyl, alkoxycarbonylalkyl having 1 to 6 carbon atoms in both the alkoxy moiety and the alkyl moiety, alkoxy-CO-NH- having 1 to 6 carbon atoms in the alkoxy moiety, an alkyl-CO-NH- group having 1 to 6 carbon atoms in the alkyl moiety, a cyano group, a heterocycloalkyl group having 5 to 9 carbon atoms, an amino-CO-NH- group, an alkylamino-CO-NH- group having 1 to 6 carbon atoms in alkyl moieties, (alkyl-aminoCO-NH- group having 1 to 6 carbon atoms in each alkyl moiety, arylamino-CO-NH group having 6 to 10 carbon atoms in the aryl moiety, heteroarylamino-CO-NH- group containing 5 to 9 carbon atoms in the heteroarylamine moiety, alkylamino-CO-NH-alkyl having 1 to 6 carbon atoms in each alkyl moiety, (alkyl-amino-CO-NH-alkyl having 1 to 6 carbon atoms in each alkyl moiety, arylamino-CO-NH-alkyl containing 1 to 6 carbon atoms in the alkyl portion and 6 to 10 carbon atoms in the aryl portion, a heteroarylamino-CO-NH-alkyl group having 1 to 6 carbon atoms in the alkyl portion and 5 to 9 carbon atoms in the heteroarylamino portion, an alkylsulfonyl group containing 1 to 6 6 carbon atoms, (C 1 -C 6) alkylsulfonylamino, (C 1 -C 6) alkylsulfonylaminoalkyl, (C 6 -C 10) arylsulfonyl group, (C 6 -C 10) arylsulfonylamino (C 6 -C 10) arylsulfonylamino (C 6 -C 10) arylsulfonylamino group 10 carbon atoms in the aryl part and 1 to 6 carbon atoms in the alkyl part, an alkylsulfonylamino group containing 1 to 6 carbon atoms, an alkylsulfonylaminoalkyl group having 1 to 6 carbon atoms in each alkyl moiety, a heteroaryl group having 5 to 9 carbon atoms, and a heterocycloalkyl group having 2 to 9 carbon atoms.

The disclosure also relates to a process for preparing stereospecific pharmaceutically acceptable acid addition salts of the compounds of formula (1) above. The acids used to prepare the pharmaceutically acceptable acid addition salts of the aforementioned basic compounds are those which form non-toxic acid addition salts, i.e. salts containing pharmacologically acceptable anions such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate , hydrogen sulphate, phosphate, hydrogen phosphate, acetate, lactate, citrate, hydrogen citrate, tartrate, hydrogen tartrate, succinate, maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate [i.e. 1,1'-methylene-bis- (2-hydroxy-3-naphthoate) salts.

The present disclosure also relates to stereospecific base addition salts of the compounds of formula I. The bases which can be used as reagents for the preparation of pharmaceutically acceptable acid addition bases of the compounds of formula I are those which form non-toxic base salts with with these compounds. Such non-toxic basic salts include those derived from pharmacologically acceptable cations such as alkali metal cations (e.g., potassium or sodium cations) and alkaline earth metal cations (e.g., calcium and magnesium cations), ammonium salts or amine addition salts. water-soluble salts such as N-methylglucamine (meglumine), as well as lower alkanolammonium salts and other basic salts of pharmaceutically acceptable organic amines, however, the scope of the present invention is not limited thereto.

The term &amp; quot; alkyl &amp; quot; as used herein, unless otherwise indicated, refers to straight or branched chain saturated monovalent hydrocarbon radicals or combinations thereof.

The term "alkoxy" as used herein refers to O-alkyl groups in which the term "alkyl" has the same meaning as previously defined.

The term &amp; quot; halogen &amp; quot; as used herein means, unless otherwise indicated, a fluorine, chlorine, bromine or iodine atom.

The compounds may contain double bonds. When such bonds are present, the compounds exist in the form of cis and trans configurations and as mixtures of these forms. Unless otherwise indicated, the alkyl and alkenyl groups mentioned herein, as well as the alkyl radicals of other groups referred to (such as alkoxy), may be linear or branched, and may also be cyclic (such as cyclopropyl). group, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl) or may be linear or branched and may contain cyclic moieties. Unless otherwise indicated, halogen means fluorine, chlorine, bromine and iodine.

As used herein, a C 2 -C 9 heterocycloalkyl group refers to pyrrolidinyl, tetrahydrofuryl, dihydrofuryl, tetrahydropyranyl, pyranyl, thiopyranyl, aziridinyl, oxiranyl, methylenedioxyl, chromenyl, isoxazolidinyl, 1,3-oxazolidin-3-yl, isothiazolidinyl, 1,3-thiazolidin-3-yl, 1,2-pyrazolidin-2-yl, 1,3-pyrazolidin-1-yl, piperidinyl, thiomorpholinyl, 1,2-tetrahydrothiazin-2-yl,

-6GB 304366 B6

1,3-tetrahydrothiazin-3-yl, tetrahydrothiadiazinyl, morpholinyl, 1,2-tetrahydrodiazin-2-yl, 1,3-tetrahydrodiazin-1-yl, tetrahydroazepinyl, piperazinyl, chromanyl, and more similar groups. It will be appreciated by those skilled in the art that the attachment of these heterocycloalkyl rings of 2 to 9 carbon atoms is via carbon or carbon atoms.³ of a hydrogenated nitrogen heteroatom.

As used herein, the term C 5 -C 9 heteroaryl refers to furyl, thienyl, thiazolyl, pyrazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl, pyrrolyl, triazolyl , tetrazolyl, imidazolyl, 1,3,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-oxadiazolyl, 1,3,5-thiadiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, 1,2,4-triazinyl, 1, 2,3-triazinyl, 1,3,5-triazinyl, pyrazolo [3,4-b] pyridyl, cinnolinyl, pteridinyl, purinyl, 6,7dihydro-5H [1] pyrindinyl, benzo [b] thiophenyl, 5,6,7,8-tetrahydroquinolin-3-yl, benzoxazolyl, benzothiazolyl, benzisothiazolyl, benzisoxazolyl, benzimidazolyl, thianaphthenyl, isothianaphthenyl, benzofuryl, isobenzofuryl, isoindolyl, indolyl, indolizinyl, quinazolyl, isoquinolines, isoquinolines, , quinazolinyl, benzoxazinyl, and the like. It will be appreciated by those skilled in the art that the attachment of these C 5 -C 9 heterocycloalkyl rings is via carbon or carbon atoms.³ hybridized nitrogen heteroatom.

As used herein, the term "C 6 -C 10 aryl" refers to phenyl or naphthyl.

Compounds used include all conformational isomers (such as cis or trans isomers). The compounds used have asymmetric centers and are therefore chiral and exist in various enantiomeric and diastereomeric forms. The disclosure relates to the resolution of optical isomers and stereoisomers of prodrugs of the ingredients and compounds and mixtures thereof, and of any pharmaceutical compositions and methods of treatment that may use or contain such agents. In this regard, both E and Z configurations are included in the description. The above compounds of formula I may also exist in the form of tautomers. These tautomers and mixtures thereof are also included. Specifically, the racemic mixture of the enantiomeric mixtures of the compounds used to form the R substituents is¹ in a compound of formula I, by treating the racemic mixture with a specific optical isomer of disubstituted tartaric acid or tartrate in a suitable solvent, such as ethanol, with or without the use of water as a co-solvent (co-solvent). A process using such resolving agents such as optical tartaric acid isomers and tartaric acid derivatives such as di-p-toluoyl-L-tartaric acid and (S) - (+) - andenoic acid salt (pencyphos, ( S) - (+) - 2-hydroxy-5,5-dimethyl-4-phenyl-1,3,2-dioxyphosphorinan-2-oxide salt), cleavage can be carried out to give the desired enantiomer in excess of 90% .

Interaction between the antipodes of the resolved substance and the specific enantiomer results in resolution of the racemic mixture, wherein one of the desired stereospecific materials is present in the precipitate of the resolved material and the enantiomers, and the remaining enantiomer present in solution can be isolated. It follows that, depending on the specific enantiomers desired and the separation method used (i.e., separation from the precipitate or solution), the stereospecific form and the cleaved form, for example the "L" form of the cleaved reagent, such as form a tartrate, giving a precipitate of the "R" form of said R substituent and a solution containing the "L" form and vice versa.

-7EN 304366 B6

The above cleavage agents are effective in obtaining the 3R, 4R enantiomer of a compound of the present invention (either in precipitate or in solution, as mentioned above):

<img file="CZ304366B6_D0005.tif" />

The cleavage method of the compound of formula 111 is carried out by the following steps:

(a) mixing the racemic mixture of a compound of formula III in a suitable solution with a cleavage compound having a defined stereospecificity for a time sufficient to achieve substantial precipitation of the stereospecific isomer of the racemic mixture from solution, (b) depending on the stereospecific form of the desired compound a precipitate which is then purified or the mother liquor is collected and the enantiomer contained in the liquor is recrystallized.

With some materials, a clot rather than a solution is formed in the cleavage described herein, so this process involves converting the suspension to another suspension. As used herein, the term "solution" refers to both a solution and a suspension.

The temperature at which said cleavage and precipitation is carried out is preferably ambient temperature, and although clotting time is not limited in terms of efficiency, this clotting time is preferably at most about four hours. To facilitate the resolution process, it is convenient to use in the racemic mixture those enantiomers which are in a stable form, the compound of formula II being most stable in the form of an acid addition salt, such as a hydrochloride salt, as compared to the free base form, this racemic mixture of the compound is subjected to conversion as described above prior to resolution. Thus, the preparation of the hydrochloride salt of the compound of formula (II) is preferably carried out in ethanol using a small amount of toluene as co-solvent (co-solvent). Alternatively, methanol, isopropanol, acetonitrile, or tetrahydrofuran (or mixtures thereof with or without the use of water as co-solvent) may be used to prepare the salt, using co-solvents such as toluene, ethyl acetate, dichloromethane, dichloroethane. or tetrahydrofuran. Particularly preferred is the use of the hydrochloride salt, since this form provides excellent purification and enrichment of the other stereomers of the previous step.

A preferred substitution solvent to be used for resolution is ethyl acetate. Toluene, acetonitrile or heptanes are also suitable as solvents.

A preferred isolating solvent is acetone. Other solvents suitable for this purpose include, for example, isopropanol, ethanol, methyl ethyl ketone, methyl isopropyl ketone, acetonitrile, and tetrahydrofuran. These solvents can also be used as co-solvents in combination with other solvents or with water.

Suitable cleavable compounds include tartaric acid and its derivatives, such as toluoyltartaric acid or benzoyltartaric acid, as mentioned in the stereospecific structure. Other resolving compounds include adenoic acid and derivatives thereof.

-8EN 304366 B6

Optionally, a vaccine addition may be used to facilitate precipitation and recrystallization, which is advantageous to obtain a material with a higher ee value and a lower number of recrystallizations. In order to illustrate this process and efficiency, examples are set forth below.

The disclosure also relates to a process for preparing a compound of formula I:

<img file="CZ304366B6_D0006.tif" />

or a pharmaceutically acceptable salt thereof, wherein in the general formula:

R represents a group of the formula:

R⁵

Rk_N/ (CH₂)_y wherein y is 0, 1 or 2,

R⁴ is selected from the group consisting of hydrogen, alkyl of 1 to 6 carbon atoms, alkylsulfonyl of 1 to 6 carbon atoms, alkenyl of 2 to 6 carbon atoms, alkynyl of 2 to 6 carbon atoms, wherein the alkyl group, alkenyl the alkynyl group may be optionally substituted with deuterium, hydroxy, amino, trifluoromethyl, (C 1 -C 4) alkoxy, (C 1 -C 6) acyloxy, (C 1 -C 6) alkylamino (C 1 -C 6 alkylamino, C 1 -C 6 alkyl, nitro, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl) or an acylamino group having 1 to 6 carbon atoms;⁴ represents a cycloalkyl group having 3 to 10 carbon atoms, the cycloalkyl group being optionally substituted by deuterium, hydroxy, amino, trifluoromethyl, acyloxy of 1 to 6 carbon atoms, acylamino of 1 to 6 carbon atoms, alkylamino of 1 to 6 carbon atoms, (an alkyl amino group containing from 1 to 6 carbon atoms in each alkyl moiety, cyano, cyanoalkyl of 1 to 6 carbon atoms, trifluoromethylalkyl of 1 to 6 carbon atoms, nitro, nitroalkyl of 1 to 6 carbon atoms or acylamino of 1 to 6 carbon atoms,

R⁵ means a heterocycloalkyl group having 1 to 9 carbon atoms, the heterocycloalkyl groups having to be substituted by one to five carboxy, cyano, amino, deuterium, hydroxy, alkyl groups having 1 to 6 carbon atoms, alkoxy groups having 1 to 6 carbon atoms, halogens, acyl groups having 1 to 6 carbon atoms, alkylamino groups having 1 to 6 carbon atoms

-9C 304366 B6 carbon atoms, C 1 -C 6 aminoalkyl, C 1 -C 6 alkoxy-C 1 -H 4, C 1 -C 6 alkoxy-C 1 -C 6 -alkyl, C 1 -C 6 -alkylamino-CO-groups, alkenyl groups C 2 -C 6 alkynyl, C 2 -C 6 alkynyl, C 1 -C 6 alkylamino, C 1 -C 6 aminoalkyl, (C1-C6) hydroxyalkyl, (C1-C6) alkoxyalkyl and (C1-C6) alkyl, (C1-C6) acyloxyalkyl and (C1-C6) alkyl moieties, nitro groups, cyanoalkyl groups having 1 to 6 carbon atoms, haloalkyl groups having 1 to 6 carbon atoms, (C 1 -C 6) nitroalkyl, (C 1 -C 6) trifluoromethyl, (C 1 -C 6) trifluoromethyl alkyl, (C 1 -C 6) acylamino, (C 1 -C 6) acylaminoalkyl and 1-C 1 acylaminoalkyl; up to 6 carbon atoms in the alkyl moiety, alkoxyacylamino groups having 1 to 6 carbon atoms in the alkoxy moiety and 1 to 6 carbon atoms in the acyl moiety, (C1 -C6) aminoacyl groups having 1 to 6 carbon atoms in the alkyl moiety and 1 to 6 carbon atoms in the acyl moiety, alkylaminoacyl groups having 1 to 6 carbon atoms in the alkyl moiety and 1 to 6 carbon atoms in the acyl moiety. in each alkyl moiety and 1 to 6 carbon atoms in the acyl moiety, by R^I5R^I6N-CO-O-;^l5R^l6N-CO-alkyl groups having 1 to 6 carbon atoms in the alkyl moiety, alkyl-S (O)_m groups containing 1 to 3 alkyl groups;

<img file="CZ304366B6_D0007.tif" />

up to 6 carbon atoms in the alkyl moiety;^l5S (0) mR^l6N, R^l5S (0) mR^l6N-alkyl groups containing up to 6 carbon atoms in the alkyl moiety, wherein m is 0, 1 or 2;

R¹⁵ and R¹⁶ each independently selected from the group consisting of hydrogen and alkyl having 1 to 6 carbon atoms, or a group of formula II:

.12 c

<img file="CZ304366B6_D0008.tif" />

<img file="CZ304366B6_D0009.tif" />

(II) / wherein a is 0, 1, 2, 3 or 4, b, c, e, f and g are each independently 0 or 1, d is 0, 1, 2 or 3,

X is S (O)_nin which n is 0, 1 or 2, an oxygen atom, a carbonyl group or a -C (= N-cyano) - group,

Y represents a S (O) n group in which n is 0, 1 or 2, or a carbonyl group, and

- 10GB 304366 B6

Z is carbonyl, C (O) O-C (O) NR- or S (O)_nwherein n is 0, 1 or 2,

R⁶, R⁷, R⁸, R⁹, R¹⁰ and R¹¹ are independently selected from the group consisting of hydrogen and alkyl of 1 to 6 carbon atoms, optionally substituted with deuterium, hydroxy, amino, trifluoromethyl, acyloxy of 1 to 6 carbon atoms, acylamino of 1 to 6 carbon atoms, alkylamino (C 1 -C 6) -alkyl group₂(C1 -C6) amino group in each alkyl moiety, cyano, (C1 -C6) cyanoalkyl, (C1 -C6) trifluoromethyl alkyl, nitro, (C1 -C6) nitroalkyl, and acylamino group 1 to 6 carbon atoms,

R is carboxy, cyano, amino, oxo, deuterium, hydroxy, trifluoromethyl, C 1 -C 6 alkyl, C 1 -C 6 trifluoromethyl, C 1 -C 6 alkoxy, halogen, (C1-C6) acyl, (C1-C6) alkylamino, (alkyl)₂an amino group having 1 to 6 carbon atoms in each alkyl moiety, an aminoalkyl group having 1 to 6 carbon atoms, an alkoxy-CO-NH group having 1 to 6 carbon atoms in the alkoxy moiety, an alkylamino-CO- group having 1 to 6 carbon atoms in the alkyl moiety, a C 2 -C 6 alkenyl group, a C 2 -C 6 alkynyl group, a C 1 -C 6 alkylamino group, a C 1 -C 6 hydroxyalkyl group, (C 1 -C 6) alkoxyalkyl, C 1 -C 6 acyloxyalkyl, both C 1 -C 6 acyl and C 1 -C 6 alkyl, C 1 -C 6 cyanoalkyl , (C 1 -C 6) haloalkyl, (C 1 -C 6) nitroalkyl, trifluoromethyl, C 1 -C 6 trifluoromethyl alkyl, C 1 -C 6 acylamino, C 1 -C 6 acylaminoalkyl, both C 1 -C 6 alkyl and C 1 -C 6 alkoxyacylamino - in both the acyl moiety, an aminoacyl group having 1 to 6 carbon atoms, an aminoacyl group having 1 to 6 carbon atoms in both the acyl and alkyl moieties (C)-C až) alkylaminoacyl group (alkyl)₂an aminoacyl group having 1 to 6 carbon atoms in each alkyl moiety and 1 to 6 carbon atoms in the acyl moiety, R^I5R¹⁶N-CO-O-;¹⁵R¹⁶An N-CO-alkyl group having from 1 to 6 carbon atoms in the alkyl moiety;^ISC (O) NH;^I5OC (O) NH, R¹⁵NHC (O) NH, alkyl-S (O)_m containing 1 to 6 carbon atoms in the alkyl moiety, alkyl 1-S (O)_m—Alkyl containing 1 to 6 carbon atoms in each alkyl moiety, R &amp; lt; 1 &amp; gt;^l5R¹⁶NS (O) m, R.^l5R^l6NS (O) m-alkyl containing 1 to 6 carbon atoms in the alkyl moiety, R^l5S (O)_mR^l6N, R^,5S (O)_mR^l6N-alkyl having 1 to 6 carbon atoms in the alkyl portion wherein m is 0, 1 or 2, and R is¹⁵ and R¹⁶ are each independently selected from the group consisting of hydrogen and alkyl of 1 to 6 carbon atoms,

R² and R³ each independently selected from the group consisting of hydrogen, deuterium, amino, halogen, hydroxy, nitro, carboxy, alkenyl of 2 to 6 carbon atoms, alkynyl of 2 to 6 carbon atoms, trifluoromethoxy, alkyl of 1; C 1 -C 6 alkoxy, C 1 -C 6 alkoxy, C 3 -C 10 cycloalkyl, wherein the alkyl group, the alkoxy or cycloalkyl group may be optionally substituted with one to three substituents selected from the group consisting of halogen, hydroxy, carboxy, C 1 -C 5 aminoalkylthio, C 1 -C 8 alkylamino, (alkyl)₂an amino group having 1 to 6 carbon atoms in each alkyl moiety, a heteroaryl group having 5 to 9 carbon atoms, a heterocycloalkyl group having 2 to 9 carbon atoms, a cycloalkyl group having 3 to 9 carbon atoms, or an aryl group having 6 to 10 carbon atoms,

Or R² and R³ each independently represent a C 3 -C 10 cycloalkyl group, a C 3 -C 10 cycloalkoxy group, a C 1 -C 6 alkylamino group, (alkyl)₂an amino group having 1 to 6 carbon atoms in each alkyl moiety, an arylamino group having 6 to 10 carbon atoms, an alkylthio group having 1 to 10 carbon atoms, an arylthio group having 6 to 10 carbon atoms, an alkylsulfinyl group having 1 to 6 carbon atoms, (C 6 -C 10), (C 1 -C 6) alkylsulfonyl, (C 6 -C 10) arylsulfonyl, (C 1 -C 6) acyl group, (C 1 -C 6) alkoxyCO-NH group, C 1 -C 6 alkylamino-CO- group, C 5 -C 9 heteroaryl, heterocycloalkyl C 2 -C 9 or C 6 -C 10 aryl, wherein the heteroaryl group, heterocycloalkyl and aryl are optionally substituted with one to three substituents selected from the group consisting of halogen, alkyl of 1 to 6 carbon atoms, alkyl-CO-NH-group of 1 to 6 carbon atoms in the alkyl moiety, alkoxy-CO-NH- (C 1 -C 6) alkoxy moiety, (C 1 -C 6) alkyl-CO-NH-alkyl (C 1 -C 6 alkyl), C 1 -C 6 -alkyl (C 1 -C 6) alkyl group as in the alkoxy moiety; in the alkyl moiety, an alkoxy-CO-NH-alkoxy group having 1 to 6 carbon atoms in each alkoxy moiety, a carboxy group, a carboxyalkyl group having 1 to 6 carbon atoms in the alkyl moiety, a carboxyalkoxy group having 1 to 6 carbon atoms in the alkoxy moiety a benzyloxycarbonylalkoxy group having 1 to 6 carbon atoms in the alkoxy moiety, an alkoxycarbonylalkoxy group having 1 to 6 carbon atoms in each alkoxy moiety, an aryl group having 6 to 10 carbon atoms, an amino group, (C 1 -C 6) aminoalkyl, (C 1 -C 6) alkoxycarbonylamino, (C 1 -C 6) arylalkoxycarbonylamino (C 6 -C 10) aryl, (C 1 -C 6) alkylamino 6 carbon atoms, (alkyl)₂an amino group having 1 to 6 carbon atoms in each alkyl moiety, an alkylaminoalkyl group having 1 to 6 carbon atoms in each alkyl moiety, (alkyl)₂(C 1 -C 6) aminoalkyl, hydroxy, (C 1 -C 6) alkoxy, carboxy, (C 1 -C 6) carboxyalkyl, (C 1 -C 6) alkoxycarbonyl, alkoxycarbonylalkyl having 1 to 6 carbon atoms in both the alkoxy moiety and the alkyl moiety, alkoxy-CO-NH- having 1 to 6 carbon atoms in the alkoxy moiety, an alkyl-CO-NH- group having 1 to 6 carbon atoms in the alkyl moiety, a cyano group, a heterocycloalkyl group having 5 to 9 carbon atoms, an amino-CO-NH group, an alkylamino-CO-NH- group having 1 to 6 carbon atoms in alkyl moieties, (alkyl)₂an aminoCO-NH- group having 1 to 6 carbon atoms in each alkyl moiety, an arylamino-CO-NH group having 6 to 10 carbon atoms in the aryl moiety, a heteroarylamino-CO-NH- group having 5 to 9 carbon atoms in the heteroarylamino moiety, alkylamino- CO — NH — alkyl having 1 to 6 carbon atoms in each alkyl moiety, (alkyl)₂amino-CO-NH-alkyl having 1 to 6 carbon atoms in each alkyl moiety, arylamino-CO-NH-alkyl having 1 to 6 carbon atoms in the alkyl moiety and 6 to 10 carbon atoms in the aryl moiety, heteroarylamino-CO- NH-alkyl having 1 to 6 carbon atoms in the alkyl moiety and 5 to 9 carbon atoms in the heteroarylamino moiety, alkylsulfonyl having 1 to 6 carbon atoms, alkylsulfonylamino having 1 to 6 carbon atoms, (C 1 -C 6) alkylsulfonylaminoalkyl, (C 6 -C 10) arylsulfonyl, (C 6 -C 10) arylsulfonylamino, (C 6 -C 10) arylsulfonylaminoalkyl (C 1 -C 6) alkyl a moiety, an alkylsulfonylamino group having 1 to 6 carbon atoms, an alkylsulfonylaminoalkyl group having 1 to 6 carbon atoms in each alkyl moiety, a C 5 -C 9 heteroaryl group and a C 2 -C 9 heterocycloalkyl group,

- 12GB 304366 B6, said process comprising the following steps:

(a) mixing a racemic mixture of enantiomeric compounds of formula (I) ₄ /^RR \_N/ (CH₂)_y

H in which y, R⁴ and R⁵ have the same meaning as above in a solvent with a cleavage compound having a defined stereospecificity to form a solution, said cleavage agent being capable of binding to at least one but not all of said isomers to form a precipitate containing said at least one of said enantiomers (b) allowing the mixture to stand for a time sufficient to achieve substantial precipitation of the stereospecific enantiomers from the racemic mixture from the solution, (c) depending on the desired stereospecific enantiomers of the compound, either a precipitate is collected and the precipitate is purified, or a solution containing the other enantiomer is collected and the enantiomer contained in the solution is recrystallized, and (d) reacting the stereospecific enantiomer thus obtained with a compound of formula XVI:

<img file="CZ304366B6_D0010.tif" />

in which

R is hydrogen or a protecting group, and R is² and R³ have the same meaning as previously defined.

The disclosure also relates to a compound of formula

<img file="CZ304366B6_D0011.tif" />

in which

- 13 GB 304366 B6

R² and R³ each independently selected from the group consisting of hydrogen, deuterium, amino, halogen, hydroxy, nitro, carboxy, alkenyl of 2 to 6 carbon atoms, alkynyl of 2 to 6 carbon atoms, trifluoromethyl, trifluoromethoxy, alkyl (C 1 -C 6) group, (C 1 -C 6) alkoxy, (C 3 -C 10) cycloalkyl group; wherein the alkyl, alkoxy or cycloalkyl group may be optionally substituted with one to three substituents selected from the group consisting of halogen, hydroxy, carboxy, aminoalkylthio of 1-6 carbon atoms, alkylamino of 1-6 carbon atoms, (alkyl)₂an amino group having 1 to 6 carbon atoms in each alkyl moiety, a heteroaryl group having 5 to 9 carbon atoms, a heterocycloalkyl group having 2 to 9 carbon atoms, a cycloalkyl group having 3 to 9 carbon atoms, or an aryl group having 6 to 10 carbon atoms, or R² and R³ each independently represent a C 3 -C 10 cycloalkyl group, a C 3 -C 10 cycloalkoxy group, a C 1 -C 6 alkylamino group, (alkyl)₂an amino group having 1 to 6 carbon atoms in each alkyl portion, an arylamino group having 6 to 10 carbon atoms, an alkylthio group having 1 to 6 carbon atoms, an arylthio group having 6 to 10 carbon atoms, an alkylsulfinyl group having 1 to 6 carbon atoms, (C 6 -C 10), (C 1 -C 6) alkylsulfonyl, (C 6 -C 10) arylsulfonyl, (C 1 -C 6) acyl group, (C 1 -C 6) alkoxy-CO-NH group, (C 1 -C 6) alkylamino-CO- group, (C 5 -C 9) heteroaryl group a (C 2 -C 9) heterocycloalkyl or (C 6 -C 10) aryl group wherein the heteroaryl group, heterocycloalkyl and aryl are optionally substituted with one to three substituents selected from halogen, alkyl of 1 to 6 carbon atoms, alkyl-CO-NH-group of 1 to 6 carbon atoms in the alkyl moiety, alkoxy-CO-NH- (C 1 -C 6) alkoxy moiety, (C 1 -C 6) alkyl-CO-NH-alkyl (C 1 -C 6 alkyl); in the alkyl moiety, an alkoxy-CO-NH-alkoxy group having 1 to 6 carbon atoms in each alkoxy moiety, a carboxy group, a carboxyalkyl group having 1 to 6 carbon atoms in the alkyl moiety, a carboxyalkoxy group having 1 to 6 carbon atoms in the alkoxy moiety a benzyloxycarbonylalkoxy group having 1 to 6 carbon atoms in the alkoxy moiety, an alkoxycarbonylalkoxy group having 1 to 6 carbon atoms in each alkoxy moiety, an aryl group having 1 to 6 carbon atoms, an amino group, (C 1 -C 6) aminoalkyl, (C 1 -C 6) alkoxycarbonylamino, (C 1 -C 6) arylalkoxycarbonylamino (C 6 -C 10) aryl, (C 1 -C 6) -alkylalkoxycarbonylamino 1 to 6 carbon atoms, (alkyl)₂an amino group having 1 to 6 carbon atoms in each alkyl moiety, an alkylaminoalkyl group having 1 to 6 carbon atoms in each alkyl moiety, (alkyl)₂(C 1 -C 6) aminoalkyl, hydroxy, (C 1 -C 6) alkoxy, carboxy, (C 1 -C 6) carboxyalkyl, (C 1 -C 6) alkoxycarbonyl, alkoxycarbonylalkyl having 1 to 6 carbon atoms in both the alkoxy moiety and the alkyl moiety, alkoxy-CO-NH having 1 to 6 carbon atoms in the alkoxy moiety, C 1 -C 6 alkyl-CO-NH-, cyano, C 5 -C 9 heterocycloalkyl, amino-CO-NH, C 1 -C 6 alkylamino-CO-NH- , (alkyl)₂an amino-CO-NH- group having 1 to 6 carbon atoms in each alkyl moiety, an arylamino-CONH- group having 6 to 10 carbon atoms in the aryl moiety, a heteroarylamino-CO-NH- group having 5 to 9 carbon atoms in the heteroarylamino moiety, an alkylamino-CO-NH-alkyl group having 1 to 6 carbon atoms in each alkyl moiety, (alkyl)₂amino-CO-NH-alkyl having 1 to 6 carbon atoms in each alkyl moiety, arylamino-CO-NH-alkyl having 1 to 6 carbon atoms in the alkyl moiety and 6 to 10 carbon atoms in the aryl moiety, hetero- 14CZ 304366 B6 arylamino-CO-NH-alkyl having 1 to 6 carbon atoms in the alkyl moiety and 5 to 5 carbon atoms in the heteroarylamino moiety, alkylsulfonyl having 1 to 6 carbon atoms, alkylsulfonylamino having 1 to 6 carbon atoms, (C 1 -C 6) alkylsulfonylaminoalkyl, (C 6 -C 10) arylsulfonyl, (C 6 -C 10) arylsulfonylamino, (C 6 -C 10) arylsulfonylaminoalkyl, (C 1 -C 6) alkyl a moiety, an alkylsulfonylamino group having 1 to 6 carbon atoms, an alkylsulfonylaminoalkyl group having 1 to 6 carbon atoms in each alkyl moiety, C 5 -C 9 heteroaryl and C 2 -C 9 heterocycloalkyl.

The present invention relates to a compound which is 3 - {(3R, 4R) -4,1-methyl-3- [methyl- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino] piperidin-1- yl) -3-oxopropionitrile.

The present invention also relates to a pharmaceutical composition for (a) treating or preventing a disorder or condition selected from the group consisting of organ transplant rejection, xenotransplantation, lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, type I diabetes and diabetes complications, cancer, asthma , atopic dermatitis, autoimmune thyroid disorders, ulcerative colitis, Crohn's disease, Alzheimer's disease, leukemia, and other autoimmune diseases, or (b) inhibiting protein kinases or Janus kinase 3 (JAK3) in a mammal, including a human, comprising a proportion of the above-described and specifically preferred compound or a pharmaceutically acceptable salt thereof, effective in such disorders or conditions, and further pharmaceutically an acceptable carrier.

The present invention also relates to a method of inhibiting protein tyrosine kinases or Janus kinase 3 (JAK3) in a mammal, including a human, comprising administering to said mammal an effective amount of the aforementioned and specifically preferred compound or a pharmaceutically acceptable salt thereof.

The present invention enables treatment or prevention of a disorder or condition selected from the group consisting of organ transplant rejection, xenotransplantation, lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, type I diabetes and complications of diabetes, cancer, asthma, atopic dermatitis, autoimmune thyroid disorders, ulcerative colitis, Crohn's disease, Alzheimer's disease, leukemia, and other autoimmune diseases in a mammal, including humans, wherein the treatment or prevention of a disorder or condition is characterized in that the mammal is administered an amount of the above-described and specified preferred compound or a pharmaceutically acceptable salt thereof, effective in such disorder or condition.

Also described herein is a compound of formula:

H₃C._ / x. 2KC!

The description also includes a compound of the formula:

him, oh₃c ch₃

- 15 GB 304366 B6

The disclosure also relates to a compound of formula:

<img file="CZ304366B6_D0012.tif" />

<img file="CZ304366B6_D0013.tif" />

COOH

HC-o '

O-CH

COOH

CH₃

The following reaction schemes illustrate the preparation of compounds. Unless otherwise indicated, R symbols², R³, R⁴ and R⁵ in these reaction schemes and in the following discussion have the same meaning as above.

Preparation

<img file="CZ304366B6_D0014.tif" />

- 16GB 304366 B6

Preparation

<img file="CZ304366B6_D0015.tif" />

(xxi)

<img file="CZ304366B6_D0016.tif" />

<img file="CZ304366B6_D0017.tif" />

(XVI)

- 17GB 304365 B6 (XVII)

Scheme 1

<img file="CZ304366B6_D0018.tif" />

<img file="CZ304366B6_D0019.tif" />

- 18 GB 304366 B6

Scheme 2

<img file="CZ304366B6_D0020.tif" />

(XX) XXIV

NR⁴R⁵ r2

<img file="CZ304366B6_D0021.tif" />

(XXJI1)

NFÚR⁵ r&gt; 2

<img file="CZ304366B6_D0022.tif" />

(XV)

- 19GB 304366 B6

Scheme 3

<img file="CZ304366B6_D0023.tif" />

In the process of Preparation A, in Reaction 1, the 4-chloropyrrolo [2,3-d] pyrimidine compound of formula XXI in which R is hydrogen or a protecting group such as benzenesulfonyl or benzyl is converted to 4-chloro A 5-halopyrrolo [2,3-d] pyrimidine compound of formula XX wherein Y is chlorine, bromine or iodine by reacting a compound of formula XXI with N-ch chlorosuccinimide, N-bromosuccinimide or N-iodosuccinimide. The reaction mixture is refluxed in chloroform for a period of from about 1 to about 3 hours, preferably for about 1 hour. Alternatively, in Reaction 1 of Preparation A, the 4-chloropyrrolo [2,3-d] pyrimidine compound of formula XXI in which R is hydrogen is converted to the corresponding 4-chloro-5-nitropyrrolo [2,3-b] pyrimidine compound wherein Y is nitro by reacting a compound of Formula XXI with nitric acid in sulfuric acid at a temperature in the range of about -10 ° C to about 10 ° C, preferably at about 0 ° C, wherein the reaction is run for a time ranging from about 5 to about 15 minutes, preferably about 10 minutes. The compound of formula XXI in which Y is nitro is converted to the corresponding 4-chloro-5-aminopyrrolo [2,3-d] pyrimidine compound of formula XX in which Y is amino by reacting the compound of formula XXI under various conditions which are well known to those skilled in the art, such as hydrogenolysis using palladium or the use of tin chloride and hydrochloric acid.

In the preparation of process A, reaction 4 of said 4-chloro-5-halopyrrolo [2,3-d] pyrimidine compound of formula XX in which R is hydrogen is converted to the corresponding compound of formula XIX in which R is alkyl a (C 1 -C 6) group or a benzyl group by treating the compound of formula (XX) with N-butyllithium at a temperature of about -78 ° C, followed by reaction of the dianionic thus obtained

The intermediate with an alkyl halide or benzyl halide at a temperature in the range of about -78 ° C to room temperature, preferably at room temperature. Alternatively, the dianionic compound so obtained is reacted with molecular oxygen to give the corresponding 4-chloro-5-hydroxypyrrolo [2,3-d] pyrimidine compound of formula XIX, wherein R is hydroxy. A compound of formula XX in which Y is bromine or iodine and R is a benzenesulfonate group is converted to a compound of formula XIX in which R is a C 6 -C 12 aryl or vinyl group by treating the compound of formula XX with N -butyllithium at a temperature of about -78 ° C, followed by the addition of zinc chloride at a temperature of about -78 ° C. The organo-zinc intermediate compound thus obtained is then reacted with an aryl iodide or vinyl iodide in the presence of a catalytic amount of palladium. The reaction mixture is stirred at a temperature in the range of about 50 to about 80 ° C, preferably at about 70 ° C, for a period of from about 1 to about 3 hours, preferably for about 1 to about 3 hours. clock.

In the preparation of Preparation A, in reaction 3, said compound of formula XIX is converted to the corresponding compound of formula XVI by treating a compound of formula XIX with N-butyllithium, lithium diisopropylamine or sodium hydride at about -78 ° C in the presence of a polar aprotic solvent such as for example tetrahydrofurari. The anionic intermediate is then reacted with (a) an alkyl halide or benzyl halide at a temperature in the range of about -78 ° C to room temperature, preferably at about -78 ° C, if R³ is alkyl or benzyl; (b) an aldehyde or ketone at a temperature in the range of about -78 ° C to room temperature, preferably at about -78 ° C, if R³ is alkoxy; and (c) zinc chloride at a temperature in the range of about -78 ° C to room temperature, preferably at about -78 ° C, and the corresponding organo-zinc intermediate compound so obtained is then reacted with aryl iodide or vinyl iodide in the presence of a catalytic amount of palladium. The resulting reaction mixture is then stirred at a temperature in the range of about 50 to about 80 ° C, preferably at about 70 ° C, for a time in the range of about 1 to about 3 hours, preferably for about 1 hour. Alternatively, the anionic compound so obtained is reacted with molecular oxygen to give the corresponding 4-chloro-6-hydroxypyrrolo [2,3-d] pyrimidine compound of formula XVI, wherein R is³ is hydroxy.

In the preparation of Preparation B, the 4-chloropyrrolo [2,3-d] pyrimidine compound of formula XXI is reacted to the corresponding compound of formula XII in the same manner as in Process 3 above. Preparation A.

In the preparation of Preparation B, in the reaction 2, the compound of the formula XXII is converted to the corresponding compound of the formula XVI, following the same procedure as described above for the reactions 1 and 2 of the preparation process A.

In reaction 1 of Scheme 1, the 4-chloropyrrolo [2,3-d] pyrimidine compound of formula XVII is converted to the corresponding compound of formula XVI in which R is benzenesulfonyl or benzyl by treating the compound of formula XVII with benzenesulfonyl chloride, benzyl chloride or benzyl bromide in the presence of a basic compound such as sodium hydride or potassium carbonate, and a polar aprotic solvent, such as dimethylformamide or tetrahydrofuran. The reaction mixture is stirred at a temperature in the range of about 0 to about 70 ° C, preferably at a temperature of about 30 ° C, for a time period between about 1 to about 3 hours, preferably about 2 hours.

-21 GB 304366 B6

In Reaction 2 of Reaction Scheme 1, a 4-chloropyrrolo [2,3-d] pyrimidine compound of formula XVI is converted to the corresponding 4-aminopyrrolo [2,3-d] pyrimidine compound of formula XV by addition of the compound of formula XVI with an amine of formula HNR⁵R⁶. This reaction is carried out in water or an alcohol as a solvent, such as tert-butanol, methanol or ethanol, or other high boiling organic solvents, such as dimethylformamide, triethylamine, 1,4-dioxane, or the like. 1,2-dichloroethane, at a temperature in the range of about 60 to about 120 ° C. Typical reaction times range from about 2 to about 100 hours, preferably about 48 hours. In case R⁵ represents a nitrogen-containing heterocycloalkyl group, then each nitrogen atom must be protected with a protecting group such as a benzyl group. Removal of R⁵ The protecting groups are carried out under conditions appropriate to the particular protecting group used, which does not affect the protecting group on the pyrrolo [2,3-d] pyrimidine ring. Removal of R⁵ protecting groups such as benzyl groups are carried out in an alcoholic solvent such as ethanol in the presence of hydrogen and a catalyst such as palladium hydroxide on carbon at a temperature ranging from room temperature to about 70 ° C. This heterocycloalkyl group containing R @ 1⁵ the nitrogen may then be further reacted with a different electrophilic agent of formula II. In the case of the preparation of urea, an electrophilic compound of formula II, such as isocyanate, carbamate and carbamoyl chloride, is reacted with R⁵ nitrogen of said heteroalkyl group in a solvent such as acetonitrile or dimethylformamide in the presence of a base such as sodium carbonate or potassium carbonate at a temperature in the range of about 20 to about 100 ° C, with a reaction time of about 24 within about 72 hours. In the case of the preparation of an amide and a sulfonamide, an electrophilic compound of formula II, such as acyl chloride and sulfonyl chloride, is reacted with R 2.⁵ nitrogen of said heteroalkyl group in a solvent such as methylene chloride in the presence of a base such as pyridine, the process being carried out at ambient temperature for a time period ranging from about 12 to about 24 hours. The amide can also be prepared by reacting a carboxylic acid with a heteroalkyl group in the presence of a carbodiimide such as 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide, in a solvent such as methylene chloride, and at ambient temperature for a period of time. about 12 to about 24 hours, or with an activated ester, such as an N-hydroxysuccinimide ester or 4-nitrophenyl ester, in a solvent, such as methylene chloride, tetrahydrofuran or ethanol. In the preparation of an alkyl compound, an electrophilic compound of formula II, such as α, β-unsaturated amide, acid, nitrile, ester and α-halamide, is reacted with R⁵ nitrogen of a heteroalkyl group in a solvent such as methanol and at ambient temperature for about 12 to about 18 hours. Alkyl formation may also be accomplished by reacting an aldehyde with a heteroalkyl group in the presence of a reducing agent such as sodium cyanoborohydride in a solvent such as methanol and at ambient temperature for about 12 to about 12 hours. 18 hours.

In Reaction 3 of Scheme 1, a protecting group is removed from a compound of formula XV when R is a benzenesulfonyl group to give the corresponding compound of formula I by treating the compound of formula XV with an alkaline base. , such as sodium hydroxide or potassium hydroxide, in an alcoholic solvent such as methanol or ethanol, or in a solvent mixture, such as an alcohol / tetrahydrofuran mixture or an alcohol / water mixture. The reaction is carried out at room temperature for a time ranging from about 15 minutes to about 1 hour, preferably 30 minutes. The deprotection of the compound of formula XV wherein R is benzyl is carried out by treating the compound of formula XV with sodium in ammonia at a temperature of about -78 ° C for about 15 minutes. minutes to about 1 hour.

In Reaction 1 of Scheme 2, a 4-chloropyrrolo [2,3-d] pyrimidine compound of formula XX is converted to the corresponding 4-aminopyrrolo [2,3-d] pyrimidine compound of formula XXVI, wherein was used as described above for Reaction 2 in Reaction Scheme 1.

In carrying out Reaction 2 in this Reaction Scheme 2, the 4-amino-5-halopyrrolo [2,3-d] pyrimidine compound of formula XXIV, wherein R is benzenesulfonate and Z is bromo or iodo, is converted to the corresponding compound of formula XXIII by reacting a compound of formula (XXIV) with (a) an aryl boronic acid, in the case where R² represents an aryl group, in an aprotic solvent such as tetrahydrofuran or dioxane, in the presence of a catalytic amount of palladium (0) at a temperature ranging from about 50 to about 100 ° C, preferably about 70 ° C, for a period of time ranging from about 2 to about 48 hours, preferably for about 12 hours; (b) alkynes, in the case where R² is an alkynyl group, in the presence of a catalytic amount of copper (I) iodide and palladium (0), and a polar solvent such as dimethylformamide, at room temperature for about 1 to about 5 hours, preferably for about 1 hour. about 3 hours; and (c) alkenes or styrenes, in the case where R 1² means a vinyl or styrenyl group, in the presence of a catalytic amount of palladium in dimethylformamide, dioxane or tetrahydrofuran, at a temperature in the range of about 80 to about 100 ° C, preferably at about 100 ° C, for a time ranging from about 2 to about 48 hours, preferably about 48 hours.

In reaction 3 of Scheme 2, the compound of formula XXIII is converted to the corresponding compound of formula XV using the procedure described for Reaction 3 of Preparation A.

In carrying out Reaction 1 in Reaction Scheme 3, a compound of Formula XVII is converted to the corresponding compound of Formula I using the procedure described for Reaction 2 in Reaction Scheme 1.

The compounds of the present invention are basic in nature and are capable of forming a wide variety of different salts with various inorganic and organic acids.

Although these salts must be pharmaceutically acceptable when administered to animals, in many cases it is convenient in practice to isolate the compounds of the present invention as pharmaceutically unacceptable salts from the reaction mixture and then simply convert the salts back to the free base compound by treatment with an alkaline reagent. and thereafter converting the free base to a pharmaceutically acceptable acid addition salt. The acid addition salts of the basic compound of the present invention are readily prepared by treating the basic compound with a substantially equivalent amount of the selected mineral or organic acid in an aqueous solvent medium or in a suitable organic solvent such as acetone, methanol or ethanol. After careful evaporation of the solvent, the desired solid salt is readily obtained. The desired acid addition salt can also be precipitated from a solution of the free base compound in an organic solvent by adding a suitable mineral or organic acid to the solution.

The compounds of the present invention, which are acidic in nature, are capable of forming base salts with various pharmacologically acceptable cations. Examples of such salts are, for example, the alkali metal or alkaline earth metal salts, with calcium, sodium and potassium salts being particularly suitable. These salts are all prepared by conventional methods. Chemical bases that are used as reagents to prepare pharmaceutically acceptable basic salts of the compounds of the present invention are those that form non-toxic base salts with the acidic compounds of the present invention. Such non-toxic basic salts include those derived from pharmacologically acceptable cations such as sodium, potassium, calcium and magnesium cations. These salts are readily prepared by treating the corresponding acidic compound with an aqueous solution containing the desired

And thereafter evaporate the solution thus obtained to dryness, preferably under reduced pressure. Alternatively, the salts may also be prepared by mixing together a lower alkanolic solution of the acidic compound and the desired alkali metal alkoxide and then evaporating the resulting solution to dryness in the same manner as above. Whichever procedure above is used. In a preferred embodiment of the invention, a stoichiometric amount of the reactants used is used to ensure that the reaction is complete and that the desired end product is maximally yielded.

The compositions of the present invention may be formulated by any of the methods well known in the art using one or more pharmaceutically acceptable carriers. Thus, the active compounds of the present invention may be formulated for oral, buccal, intranasal, parenteral (i.e., intravenous, intramuscular or subcutaneous) or rectal administration, or may be formulated for administration by inhalation or insufflation. The active compounds of the present invention may also be formulated in a sustained release formulation.

For oral administration, the pharmaceutical composition may be in the form of, for example, a tablet or capsule, prepared by conventional methods known in the art using pharmaceutically acceptable excipients such as binding agents (e.g., pregelatinized corn starch, polyvinylpyrrolidone or the like). hydroxypropylmethylcellulose), fillers (such as lactose, microcrystalline cellulose or calcium phosphate), lubricants (such as magnesium stearate, talc or silica), disintegrating agents (such as potato starch or sodium starch glycolate), or wetting agents (such as sodium luarylsulfate). The tablets may be coated according to methods well known in the art. Liquid preparations for oral administration may be in the form of, for example, a solution, syrup, or suspension, or may be prepared as a dry product to be treated with water or other suitable vehicle, prior to use. These liquid preparations may be prepared by conventional methods well known in the art using pharmaceutically acceptable additives such as suspending agents (e.g. sorbitol syrup, methylcellulose or hydrogenated edible fats), emulsifying agents (e.g. lecithin or acacia) ), non-aqueous vehicles (such as almond oil, oily esters or ethyl alcohol), and preservatives (such as methyl or propyl p-hydroxybenzoates or sorbic acid).

For buccal administration, the compositions of the present invention may take the form of tablets or lozenges which are also prepared by conventional methods.

The active compounds of the present invention may be formulated for parenteral administration by injection, including administration by catheterization or infusion. Formulations for injection may be presented in unit dosage form, such as in the form of ampoules or multidose units, with the addition of preservatives. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and / or dispersing agents. Alternatively, the active ingredients may be in powder form for reconstitution with a suitable vehicle just prior to use, such as sterile pyrogen-free water. The active ingredients of the present invention may also be formulated in rectal compositions such as suppositories or retention enemas, containing, for example, commonly used suppository bases such as cocoa butter or other glycerides.

For intranasal or inhalation administration, the active compounds of the present invention are typically administered as a solution or suspension using a spray applicator canister to dispense the active mixture by squeezing a cattle or pump, or using an aerosol can. spray, which is applied by means of a pressurized container or nebulizer using a suitable propellant (propellant), such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by means of a valve which is designed to release a metered amount. The pressurized container or nebulizer may contain a solution or suspension of the active compound. Capsules or cartridges (made using, for example, gelatin) used in an inhaler or insufflator may be formulated containing a powder mix of a compound of the present invention and a suitable powder base such as lactose or starch.

The proposed dosage of the active compounds of the present invention for oral, parenteral or buccal administration to an average adult human for the treatment of the conditions mentioned above (see, for example, rheumatoid arthritis) ranges from 0.1 to 100 milligrams of active ingredient per unit dose. be administered, for example, one to four times a day.

Aerosol formulations for the treatment of the above conditions (such as asthma) in an average adult human are preferably formulated such that each metered dose or "released" aerosol dose contains 20 to 100 µg of a compound of the present invention. The total daily dose of the aerosol will range from 0.1 to 1000 mg. Administration may be several times a day, for example two, three, four or eight times a day, for example 1, 2 or 3 doses each.

The compound of formula (I) of the present invention may be administered in pharmaceutically acceptable form either alone or in combination with one or more other agents that modulate the immune system in a mammal, or concomitantly with anti-inflammatory agents, or agents including cyclosporin A ( such as Sandimmune® or Neoral®, rapamycin, FK-506 (taclolimus), leflunomide, deoxyspergualin, mycophenolate (such as Celicept®, azathioprine (such as Imuran®), daclizumab (such as Zenapax®), OKT3 (such as Orthocolone®), AtGam, aspirin, acetaminophen, ibuprofen, naproxen, piroxicam, and anti-inflammatory steroids (such as prednisolone or dexamethasone), without limiting the scope of the present invention, however, these agents may be administered as part of the same or other separate dosage form, by the same or a different dosing method and by the same or different dosing regimens, applying the knowledge of standard pharmaceutical practice.

FK-506 (tacrolimus) is administered orally at a dose of 0.10 to 0.15 mg / kg body weight every twelve hours, the first dose being administered 48 hours after surgery. This dose is monitored as tacrolimus serum levels.

Cyclosporin A (Sandimmune, oral or intravenous formulation, or Neoral®, oral solution or capsule) is administered orally at a dose of 5 mg / kg body weight every twelve hours at 48 hours post-operation. The dose is monitored by the level of maximal decrease in cyclosporin A in the blood.

The active ingredients may be formulated in sustained release dosage forms using well known methods in the art. Examples of such formulations can be found in U.S. Patents 3,538,214, 4,060,598, 4,173,626, 3,119,742 and 3,492,397.

The ability of the compounds of Formula I of the present invention, or pharmaceutically acceptable salts thereof, to inhibit Janus kinase 3 and the resulting demonstration of their efficacy for treating disorders or conditions characterized by Janus kinase 3 is illustrated by the following in vitro assays.

-25GB 304366 B6

Biological tests

JAK3 (JH1: GST) enzyme assay

The JAK.3 kinase assay uses a protein expressed in baculovirus-infected SF9 cells (GST fusion protein and human JAK3 catalytic domains) purified by glutathione-Sepharose affinity chromatography. The substrate for the reaction is polyglutamate-tyrosine (PGT (4: 1), Sigma Catalog, No. P0257), which coated the Nunc Maxi Sorp plates at a concentration of 100 µg / ml overnight at 37 ° C. In the morning following the plate coating, the plates were washed three times and JAK.3 was added to wells containing 100 μΐ kinase buffer (50 mM HEPES, pH 7.3, 125 mM NaCl, 24 mM MgCl₂) + 0.2 μΜ ATP + 1 mM sodium orthovanadate). The reaction was continued for 30 minutes at room temperature and the plates were washed three times. The level of phosphorylated tyrosine in a given well was quantified by a standard ELISA using an anti-phosphotyrosine antibody (1CN PY20, Cat # 69-151-1).

Inhibition of IL-2 dependent human T lymphoblast proliferation

This assay measures the inhibitory effect of compounds on IL-2 dependent T cell proliferation in vitro. Since signaling through the IL-2 receptor requires JAK.-3, cellular active inhibitors of JAK-3 can inhibit IL-2 dependent T cell proliferation.

Cells for this assay were isolated from fresh human blood. After monoclonal cells were separated using Accuspin System-Histopaque-1077 (Sigma # A7054), primary human T cells were isolated by negative selection using the Lympho-Kwik T kit (One Lambda, Inc., Cat # LK-50T). . T lymphocytes were grown at a concentration of 1-2 x 10⁶/ ml in medium (RPMI + 10% heat-inactivated fetal calf serum (Hyclone cat. no. A-III-L) + 1% penicillin / streptomycin (Gibco) and induced to proliferate by adding PHA at a concentration of 10 µg / ml (Murex Diagnostics Cat. No. HA 16) After 3 days at 37 ° C in 5% CO 2₂ cells were washed in medium, resuspended to a density of 1-2 x 10 cells / ml plus 100 units / ml human recombinant IL-2 (R&amp;D Systems, Cat. No. 202-IL). After 1 week, the cells were IL-2 dependent and could be maintained for up to 3 weeks by administering the same volume of medium + 100 units / ml twice weekly.

To test the ability of the test compounds to inhibit IL-2 dependent T cell proliferation, IL-2 dependent cells were washed three times, resuspended in medium, and then plated (50,000 cells / well / 0.1 ml) in 96-well microtiter plates. flat bottom (Falcon # 353075). A 10 mM stock solution of the test compound in DMSO was added in duplicate to triplicate wells starting at a concentration of 10 µΜ. After one hour, 10 units / ml IL-2 was added to each well. Plates were then incubated at 37 ° C, 5% CO₂ for 72 hours. H-thymidine (0.5 µCi / well) (pulse) (NEN Cat. No. NET02 / A) was then added to the plates and incubated for an additional 18 hours. The culture plates were then harvested using a 96-well plate reader and quantity³H-thymidine incorporated into proliferating cells was determined by reading on a Packard Top Count scintillation counter. Data were analyzed by plotting% inhibition of proliferation versus test compound concentration. The IC value was determined from this graph₅₀ (μΜ).

DETAILED DESCRIPTION OF THE INVENTION

The following examples illustrate the preparation of the compounds of the present invention, but the scope of the present invention is not limited to the details. Melting points are uncorrected. NMR data are reported in parts per million (δ) and are related to the deuterium blocking signal from the sample solution (deuteriochloroform, unless otherwise indicated). In these examples, commercial reagents were used without further purification. THF refers to tetrahydrofuran.

-26EN 304366 B6

DMF refers to Ν, Ν-dimethylformamide. Low resolution mass spectra (LRMS) were recorded either on a Hewlett Packard 5989® instrument using chemical ionization (ammonia) or a Fisons (or Micro Mass) instrument with an atmospheric pressure chemical ionization (APCI) platform using 50 / 50 mixture of acetonitrile and water with 0.1% formic acid as ionizing agent. Room or ambient temperature means 20-25 ° C.

Example 1 (Preparation of stable salt)

(1-Benzyl-4-methyl-piperidin-3-yl) -methylamine bis hydrochloride

A solution containing 23.4 kilograms of (1-benzyl-4-methylpiperidin-3-yl) methylamine in 10 liters of toluene and 120 liters of ethanol at 3 ° C was added, followed by 25 liters of 32% acid solution. hydrochloric acid in water, and the reaction mixture was kept below 10 ° C. Then, 100 liters of solvent were distilled off under partially reduced pressure, followed by 215 liters of ethyl acetate at 30 ° C. 210 liters of solvent were distilled off under partially reduced pressure, and a further 215 liters of ethyl acetate were added thereto, and 210 liters of solvent were again distilled off under partially reduced pressure. 111 L of acetone was added at 35 ° C, the suspension was cooled to 0 ° C, and the product (1-benzyl-4-methylpiperidin-3-yl) methylamine bihydrochloride was filtered off and washed. 55 liters of acetone. The wet cake was then resuspended three times in ethanol (10 volume equivalents at reflux) to improve the diastereomeric cis: trans ratio from 91: 9 to greater than 97: 3. Total yield: 19.4 kilograms , 62%. % NMR (CD₃OD, 400 MHz):

7.55 (m, 5H), 4.88 (s, 3H), 4.52 (d, J = 12.8 Hz, 1H), 4.45 (d, J = 12.8 Hz, 1H), 3.76 (m, 1H), 3.67 (m, 1H), 3.40-3.00 (m, 3H), 2.78 (3, 3H), 2.55 (m, 1H), 2 14 (m, 1H); 1.90 (m, 1H); 1.16 (d, J = 7.2 Hz, 3H).

Example 2 - Comparative Example (Cleavage)

Preparation of bis [(1-benzyl-4-methylpiperidin-3-yl) methylamine] di-p-toluoyl-L-tartrate]

To a solution containing 9.5 kg of (1-benzyl-4-methylpiperidin-3-yl) methylamine bishydrochloride in 16 liters of water was added 33 liters of 2N sodium hydroxide solution. A proportion of solids precipitated from this mixture. The resulting suspension was diluted with 43 L of isopropanol and 11 L of methanol to redissolve the solids. Di-p-toluoyl-L-tartaric acid (6.3 kg) was then added to precipitate a solids. The suspension was heated to reflux to dissolve these solids and then slowly cooled to 72 ° C. Seeds of bis [(1-benzyl-4-methylpiperidin-3-yl) methylamine] di-p-toluoyl-L-tartrate (180 grams) were added, and the cloudy solution was slowly cooled to 15 ° C. . The solid was filtered and washed with isopropanol to yield 5.9 kg of bis [(1-benzyl-4-methylpiperidin-3-yl) methylamine] di-p-toluoyl-tartrate, representing 44%. 1 HNMR (CD 3 OD, 400 Hz):

8.04 (d, J = 8.4Hz, 2H), 7.30 (m, 7H), 5.86 (s, 1H), 4.91 (s, 3H), 3.64 (d, J) = 12.8 Hz, 1H), 3.41 (d, J = 12.8 Hz, 1H), 3.09 (s, 1), 2.90 (m, 2H), 2.40 (s, 3H) ). 2.22 (m, 2H), 1.92 (m, 1H), 1.57 (m, 2H), 1.03 (d, J = 7.2 Hz, 3H).

-27GB 304366 B6

Example 3 - Comparative Example (Phencyphos cleavage)

To a solution of 6.83 g (31.3 mmol) of IPA and 10 ml of water was added 7.57 g of (+) - phencyphos (31.3 mmol), and the mixture was heated to reflux. condenser to obtain a clear solution. Seed crystals were then added at about 65 ° C using an ee of 90%. Crystallization began within one hour and the temperature was allowed to rise to room temperature overnight. Separation yielded 6.85 g (47%) of the product at ee 99%. The filtrate was concentrated, TBME, water and K were added₂WHAT₃ and the layers were separated. The organic layer was then dried (using sodium sulfate) and the solvent was evaporated. The resulting oily product (3.99 g) was then dissolved in 200 ml IPA and 10 ml water and 4.4 g of (-) - phencyphos was added. The mixture was heated to reflux and allowed to cool to room temperature overnight. 6 g of salt (41% yield) with an ee of 99.9% were obtained. Analysis was performed with free amine. The free amine was obtained by treating this salt with TBME, water and K₂WHAT₃.

The following reaction scheme illustrates the procedures of Examples 1 to 3 (wherein Bn is benzyl -CH 2 -C₆H₅):

_ NBn

CH₃HN racemic

HCl, water, ethanol

<img file="CZ304366B6_D0024.tif" />

2HCI

H₃C",

O

1) NaOH

2) di-p-toluoyl-L-tartrate 2HCl ipo, MeOH

CH₃HN

CK₃HN 'COOH racemic o hc-o ^ Q-ch, HsC- ^ y ^ O-CH

CÓOH

99% ee

<img file="CZ304366B6_D0025.tif" />

1) free base

2) HO. 0 '%

<img file="CZ304366B6_D0026.tif" />

<img file="CZ304366B6_D0027.tif" />

-28GB 304366 B6

Example 4 - Comparative Example

The racemic mixture of the compound of formula IIIa was resolved:

<img file="CZ304366B6_D0028.tif" />

CP-673,881

Sample processing:

The compound of formula III above was filtered through a 0.2 µm nylon filter disc 66.

Procedure: (96% ethanol, 4% water as solvent)

0.8711 g of the compound of formula III, the filtrate, was dissolved in 5.0 ml of a 96: 4 mixture of ethanol and water. Then 1.544 g of di-p-toluoyl-L-tartaric acid was added and the reaction mixture was stirred to form a clear solution. This solution was then allowed to stand at room temperature for about 4 hours. The resulting suspension was then filtered on Whatman No. 2 filter paper and then washed with 4.0 mL of a 96: 4 mixture of ethanol and water. The solid was then air dried to give 0.488 g of the diastereomeric salt.

0.488 g of this diastereomeric salt was suspended in 50 ml of water and 50 ml of methylene chloride was added. The pH of the mixture was then adjusted to about 9 using a saturated sodium bicarbonate solution, followed by the addition of 1.0 N sodium hydroxide solution. Upon completion of pH adjustment, the layers were separated and the methylene chloride layer was then filtered using Whatman # 2 filter paper. The solvents were removed by evaporation under reduced pressure to give a light orange oil. Weight not determined. This oil was then analyzed by gas chromatography.

Analytical assay: 97.3% of the desired enantiomer, determined from normalized area%.

Example 5 - Comparative Example

Procedure: (100% ethanol as solvent)

0.8714 grams of (1-benzyl-4-methylpiperidin-3-yl) methylamine was dissolved in 5.0 ml proof¹of ethanol 200, then 1.544 g of di-p-toluoyl-L-tartaric acid was added and the mixture was stirred to give a clear solution. The solution was allowed to stand at room temperature for about 4 hours. The resulting suspension was filtered on Whatman # 2 filter paper, which was washed with 4.0 mL of a 96: 4 mixture of ethanol and water. The solid was air dried to yield 0.628 g of diastereomeric salt.

-29EN 304366 B6

0.628 g of the diastereomeric salt was suspended in 50 ml of water and then 50 ml of methylene chloride was added. The pH of the mixture was then adjusted to about 9 using a saturated sodium bicarbonate solution followed by a 0.1N sodium hydroxide solution. After adjusting the pH, the layers were separated and the methylene chloride layer was filtered through Whatman # 2 filter paper. The solvents were removed by evaporation under reduced pressure to give a pale yellow oil. Weight not determined. This oil was then analyzed by assay: 90.5% of the desired enantiomer, determined from normalized area%.

Example 6

3 - {(3R, 4R) -4-Methyl-3- [methyl- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino] piperidin-1-yl} 3-oxopropionate

Method A (3R, 4R) - (1-Benzyl-4-methyl-piperidin-3-yl) -methyl- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) -amine

4-Chloropyrrolo [2,3-d] pyrimidine (5.37 grams, 34.9 mmol) was prepared according to the method of Davoll, J. Am. Chem. Soc., 82, 131 (1960), the product of Example 2 (6 grams, 27.5 mmol) and potassium carbonate (11.4 grams, 82.5 mmol) were combined in water (60 mL). The suspension thus obtained was heated at reflux for 90 hours. The mixture was cooled to 90 ° C and toluene (60 mL) was added. The biphasic mixture was filtered through filter paper and the layers separated. The aqueous layer was then extracted with toluene. The toluene layers were combined and washed with 1N sodium hydroxide solution, treated with activated carbon and filtered through a filter aid. Toluene was evaporated in vacuo and the residue was crystallized from 1: 1 isopropyl acetate: hexane to give 5 g of off-white solid. Yield: 54%, LRMS: 336.1 (M + 1).

Method B

Methyl - ((3R, 4R) -4-methylpiperidin-3-yl) - {7H-pyrrolo (2,3-d) pyrimidin-4-yl) amine

To the product of Method A (0.7 g, 2.19 mmol) dissolved in 15 mL of ethanol was added 1.5 mL of 2N hydrochloric acid solution and the reaction mixture was then degassed with nitrogen purge. 0.5 grams of 20% palladium hydroxide on carbon (50% water) (Aldrich) was added to the reaction mixture, and the resulting mixture was shaken (Parr-Shaker) under 50 psi of hydrogen (50 psi). at room temperature for 2 days. The reaction mixture was filtered through celite and concentrated to dryness in vacuo, and the residue was purified by flash chromatography (silica gel; 5% methanol in dichloromethane) to give 0.48 g (90% yield). of the title compound. LRMS: 246.1 (M + 1).

Method C

3 - ((3R, 4R) -4-Methyl-3- [methyl- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino] piperidin-1-yl) -3-oxopropionitrile

0.82 g of cyanoacetic acid 2,5-dioxopyrrolidin-1-yl ester was added to a stirred solution containing the product of Method B above (1.0 g) dissolved in 30 ml of ethanol, and the resulting mixture was then stirred at room temperature for 2 hours. The reaction mixture was then filtered through Celite® and concentrated in vacuo. The residue was redissolved in dichloromethane, washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated to dryness in vacuo to give 1.1 g (86% yield) of the title compound as a white solid. yellow foam. LRMS: 313 (M + 1).