Methods for selectively inhibiting janus tyrosine kinase 3 (jak3)

A method of therapeutically treating a mammalian subject to suppress an undesired immune response, comprising

• administering to a subject experiencing or at risk of experiencing said undesired immune response a pharmaceutical composition containing, a carrier and at least one compound of the formula (I)
• wherein
• R1 is H, ═CH2, CH2N(CH3)2, CH2SC(O)CH3, CH2SC6H5, CH2SCH2-(4-C6H4OCH3), CH2SC(O)C6H5 or CH2N(CH2CH3)2;
• R2 is O;
• R3is CH2N(CH3)2, CH2N(CH2CH3)2 or CH2-(N-morphyl);
• or a pharmaceutically acceptable salt thereof, or a metabolite, or a precursor of said compound capable of being converted in the body of the subject to said at least one compound, in an amount effective to interfere with the signal 3 pathway in a lymphoid cell expressing Janus tyrosine kinase 3 (Jak3) and thereby inhibit proliferation of said lymphoid cell, to suppress said undesired immune response.

A method of inhibiting proliferation of a Jak3-dependent leukemia or lymphoma in a mammalian subject suffering from said leukemia or lymphoma, comprising

• administering to the subject a pharmaceutical composition containing a carrier and at least one compound of the formula (I)
• wherein
• R1 is H, ═CH2, CH2N(CH3)2, CH2SC(O)CH3, CH2SC6H5, CH2SCH2-(4-C6H4OCH3), CH2SC(O)C6H5 or CH2N(CH2CH3)2;
• R2 is O;
• R3is CH2N(CH3)2, CH2N(CH2CH3)2 or CH2-(N-morphyl);
• or a pharmaceutically acceptable salt thereof, or a metabolite of said compound, or a precursor of said compound capable of being converted in the body of the subject to said at least one compound, in an amount effective to selectively inhibit Jak3 activity relative to other kinase activity and thereby inhibit proliferation of said leukemia or lymphoma cell.

An in vitro method to aid in identifying a new immunosuppressive drug comprising:

• testing a compound of interest for activity for disrupting T-cell function by contacting a T-cell expressing Jak3 with said compound of interest over a range of concentrations and determining whether said compound inhibits Jak3 activity at one or more concentration within said range;
• comparing the Jak3 inhibitory activity of said compound of interest to that of the compound of the formula (I)
• wherein
• R1 is H, ═CH2, CH2N(CH3)2, CH2SC(O)CH3, CH2SC6H5, CH2SCH2-(4-C6H4OCH3), CH2SC(O)C6H5 or CH2N(CH2CH3)2;
• R2 is O;
• R3is CH2N(CH3)2, CH2N(CH2CH3)2 or CH2-(N-morphyl), having predetermined Jak3 inhibitory activity;
• and
• using the results of said testing and comparing to determine whether said compound of interest is a candidate drug for in vivo use as a therapeutic immunosuppressive agent.

An in vivo method of testing a candidate immunosuppressive drug for its effect on allograft survival comprising:

• (a) implanting an allograft taken from a suitable donor animal into a suitable recipient animal;
• (b) maintaining basic nutrition and health promoting conditions for the animals;
• (c) administering the candidate drug to at least one animal, to provide a treated recipient or group;
• (d) administering to at least one said animal a pharmaceutical composition comprising a carrier containing the compound
• wherein R1 and R3 are each CN(CH3)2 and R2 is O, to serve as a positive control group;
• (e) optionally, leaving at least one recipient animal untreated, to serve as an untreated control recipient or group;
• (f) determining allograft survival time of each allograft in each recipient;
• (g) performing a histologic examination of each allograft and assessing drug related structural damage to each allograft, as applicable;
• (h) comparing the allograft survival time and the histologic structural changes in each allograft;
• and
• (i) determining from said comparison whether allograft survival time is enhanced by said candidate drug compared to the allograft(s) from the untreated control recipient(s) or compared to the allograft(s) from the positive control recipient(s), and determining whether there is drug-induced structural damage to the candidate drug-treated allograft, wherein enhanced allograft survival time and lack of drug-induced structural damage to the allograft is indicative that the candidate drug is likely to be effective when used therapeutically in vivo as an immunosuppressive agent.

A method of inhibiting an undesired Janus tyrosine kinase 3 (Jak3)-dependent function of a cell expressing Janus tyrosine kinase 3, comprising contacting the cell with at least one compound of the formula (II)

• wherein
• n is 1,2,3,4 or 6;
• R1 is H, CH2, or CH2N(CH3)2;
• R3 is CH2N(CH3)2;
• or a salt thereof, at a concentration effective to selectively inhibit the activity of said Janus tyrosine kinase 3 in said cell.

A method of inhibiting an undesired Janus tyrosine kinase 3 (Jak3)-dependent function of a cell expressing Janus tyrosine kinase 3, comprising contacting the cell with at least one compound of the formula (III)

• wherein
• n is 1 or 2;
• R1 is H, or CH2N(CH3)2;
• R3 is CH2N(CH3)2;
• or a salt thereof, at a concentration effective to selectively inhibit the activity of said Janus tyrosine kinase 3 in said cell.

An in vivo method of selectively inhibiting Janus tyrosine kinase 3 (Jak3) activity in Jak3-expressing cells in a mammalian subject, comprising administering to said subject a pharmaceutical composition comprising a carrier and at least one compound of the formula (I)

• wherein
• R1 is H, ═CH2, CH2N(CH3)2, CH2SC(O)CH3, CH2SC6H5, CH2SCH2-(4-C6H4OCH3), CH2SC(O)C6H5 or CH2N(CH2CH3)2;
• R2 is O;
• R3is CH2N(CH3)2, CH2N(CH2CH3)2 or CH2-(N-morphyl);
• or a salt thereof, or a metabolite or said compound, or a precursor of said compound capable of being converted in the body of the subject to said at least one compound, in an amount effective to inhibit said Jak3 activity in said Jak3-expressing cells without inhibiting other Janus tyrosine kinase activities in the subject, or inhibiting said Jak3 activity at least three-fold more than other Janus tyrosine kinase activities in the subject.