JP2010522724A - Pharmaceutical composition containing cytokine - Google Patents

Abstract

The present invention relates to (i) a cytokine, and (ii) a compound of the general formula (Ih), or an acid or base and a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable prodrug, or a compound thereof. The present invention relates to a pharmaceutical composition containing a stereoisomer. Examples of cytokines include (i) four α-helix cytokine families consisting of (a) IL-2 subfamily, (b) interferon (IFN) subfamily, and (c) IL-10 subfamily, ( It is selected from the group of cytokine families of ii) IL-1 family, (iii) IL-17 family, and (iv) chemokines.
[Selection figure] None

Description

  The present invention relates to treatment, more specifically viral infections such as hepatitis B or C, influenza or HIV infection, inflammation or autoimmune diseases such as multiple sclerosis, rheumatoid arthritis, inflammatory bowel It relates to the field of treatment of diseases such as inflammatory diseases, psoriasis and reperfusion injury such as stroke or myocardial infarction and various tumors. The invention relates in particular to the therapeutic use of compositions containing cytokines and compounds of general formula (Ih).

  The present inventor has recently found unexpected beneficial effects of a compound of general formula (Ih), an acid or base and a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable prodrug, or a stereoisomer thereof. And can be used for the treatment of animal or human diseases.

  Cytokines, on the other hand, are proteins secreted from cells that mediate or regulate immunity, inflammation and hematopoiesis. They must be generated de novo in response to immune stimulation. Responses to cytokines include increasing or decreasing the expression of membrane proteins (including cytokine receptors) and secretion and proliferation of effect molecules. Various cytokines such as interferon, IL-2 or TNF-α are used as therapeutic agents in various diseases.

  The inventors have more surprisingly (i) cytokines, (ii) compounds of general formula (Ih), acids or bases and pharmaceutically acceptable salts thereof, pharmaceutically acceptable prodrugs, It has now been found that pharmaceutical compositions comprising or stereoisomers thereof have unexpected and beneficial effects. It has been found that administering a compound of general formula (Ih) with a cytokine exerts an unexpected synergistic effect in viral replication and PBMC proliferation assays. Further surprisingly, the inventor can show that the cytotoxicity of both substances has been reduced, thus showing an antagonistic cytotoxic effect. Both results expand the applicability of both substance groups, ie the compound of formula (Ih) and cytokine, if applied together, by mutual enhancement of effects and reduction of toxicity.

FIG. 1 shows anti-BVDV activity in DBK cells infected with compound 273 (see table) and INF-α MBVDV virus. Further details regarding data calculations are outlined in the section on BVDV assay-data analysis. In summary, the additional behavior of the two compounds is shown by the horizontal plane, the synergistic behavior is shown by the upper face peak, and the antagonistic behavior is shown by the lower face depression. A 95% confidence interval around the experimental dose response surface is used to statistically evaluate the data, and peaks / wells are used to quantify the amount of synergy / antagonism produced. FIG. 2 shows the anti-BVDV cytotoxicity of compound 273 and INF-α in MDBK cells infected with BVDV virus. Further details regarding data calculations are outlined in the section on BVDV assay-data analysis. In summary, the additional behavior of the two compounds is shown by the horizontal plane, the synergistic behavior is shown by the upper peak, and the antagonistic behavior is shown by the lower indentation. A 95% confidence interval around the experimental dose response surface is used to statistically evaluate the data, and peaks / wells are used to quantify the amount of synergy / antagonism produced.

Compound group of general formula (Ih) The present invention provides a compound of general formula (Ih), an acid or base and a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable prodrug, or a stereoisomer thereof. It is related with the pharmaceutical composition containing.

Where
A is NR ^{2 ′} , S or O;
t is 0 to 4;
r is 0 or 1;

R ^2a is independently H, OH, SH, NH ₂ , alkyl, cycloalkyl, hydroxyalkyl, haloalkyl, haloalkyloxy, alkoxy, alkylamino, hydroxyalkylamino, halogen, aryl, or heteroaryl;

R ^3a is H, OH, SH, NH ₂ , —C (NR ⁷ ) NR ^{7 ′} R ⁸ , — (CH ₂ ) _p aryl, — (CH ₂ ) _p NR ⁷ R ⁸ , —C (O) NR ⁷ R ⁸ , -N = CR ⁷ R ⁸ , -NR ⁷ C (O) R ⁸ , alkyl, cycloalkyl, hydroxyalkyl, haloalkyl, haloalkyloxy, alkoxy, alkylamino, hydroxyalkylamino, halogen, aryl, or Is heteroaryl;

R ^d is H, halogen, alkyl, -C (NR ⁷ ) NR ^{7 '} R ⁸ ,-(CH ₂ ) _p aryl,-(CH ₂ ) _p NR ⁷ R ⁸ , -C (O) NR ⁷ R ^{8 , -N = CR 7 R 8,} -NR 7 C (O) R 8, cycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, heteroaryl, or aryl;

R ¹ is -C (O) R ^7a , -C (O) CHR ⁷ R ⁸ , -C (O) NR ⁷ R ⁸ , -C (O) OR ⁷ , -R ⁷ C (O) R ⁸ , Or -C (S) R ^7b ;
R ² is H, alkyl, cycloalkyl, heterocycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, or heteroaryl; or
R ¹ and R ² together form a 3-8 membered saturated or at least partially unsaturated monocyclic or polycyclic system with N- or C-atoms, wherein at least one Or more carbon atoms in the ring are substituted by a heteroatom selected from O, N, and S, and the ring can be substituted by one or more R ⁹ ;

R ^4a is H, C ₁ -C ₆ -alkyl, C ₂ -C ₆ -alkenyl, cycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, -C (NR ⁷ ) NR ^{7 '} R ⁸ ,- (CH ₂ ) _p aryl,-(CH ₂ ) _p NR ⁷ R ⁸ , -C (O) NR ⁷ R ⁸ , -N = CR ⁷ R ⁸ , -NR ⁷ C (O) R ⁸ , halogen, heteroaryl Or aryl;

R ³ is H, —C (O) NR ^a R ^b , halogen, alkyl, haloalkyl, aryl, heteroaryl, OH, SH, NR ^{4 ′} OR ^{5 ′} , NH ₂ , hydroxyalkylamino, alkylamino, alkoxy, Is cycloalkyl, heterocycloalkyl, hydroxyalkyl, or haloalkyloxy;

R ^a is H, halogen, alkyl, —C (NR ⁷ ) NR ^{7 ′} R ⁸ , — (CH ₂ ) _p aryl, — (CH ₂ ) _p NR ⁷ R ⁸ , —C (O) NR ⁷ R ^{8 , -N = CR 7 R 8,} -NR 7 C (O) R 8, cycloalkyl, heterocycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, heteroaryl, or aryl;

R ^b is independently H, -CN, -OH, -SH, -CO ₂ R ^{4 '} , -C (O) R ^4' , -SO ₂ NR ^{4 '} , NR ^4' R ^{5 '} , -C ( O) NR ⁷ R ⁸ , -SO ₂ -alkyl, -SO ₂ R ^{4 '} , SO ₃ R ^4' , -N = CR ^{4 '} R ^5' , -NR ^{4 '} C (O) R ^4'' , NR ^{4 '-CO- _{haloalkyl, NO 2, -NR 4' -SO}} 2 - heteroaryl - ^'alkyl, -NR ⁴ -SO ^_2' -CO- alkyl, -NR ^{4 '(CH} ₂₎ to _p haloalkyl, -NR ⁴ , Alkyl, cycloalkyl, alkylamino, alkoxy, alkylthio, halogen, haloalkyl, haloalkyloxy, O (CH ₂ ) _p [O (CH ₂ ) _p ] _q OCH ₃ , —C (NR ^{4 ″} ) NR ^{4 ′} benzimidazolyl , C (NR ^{4 ''} ) NR ^{4 '} benzthiazolyl, -C (NR ^4'' ) NR ^4' benzoxazolyl, hydroxyalkyl, hydroxy-cycloalkyl, hydroxyalkylamino, heterocycloalkyl, aryl, or heteroaryl Represents;

R ^{4 ′} , R ^{4 ″} , R ^{5 ′} are independently H, halogen, alkyl, —C (NR ⁷ ) NR ^{7 ′} R ⁸ , — (CH ₂ ) _p aryl, haloalkyl, — (CH ₂ ) _p NR ⁷ R ⁸ , -C (O) NR ⁷ R ⁸ , -N = CR ⁷ R ⁸ , -NR ⁷ C (O) R ⁸ , cycloalkyl, heterocycloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, Is heteroaryl, or aryl;

R ⁷ , R ^{7 ′} , R ⁸ are independently H, halogen, alkyl, cycloalkyl, heterocycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, arylamino heteroaryl, or aryl;

R ^7a is cycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, heteroaryl, or aryl;

R ^7b is H, halogen, alkyl, cycloalkyl, heterocycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, heteroaryl, or aryl;

X is NR ^{2 ′} , O, or S;
Z is N or CR ^{2 ′} ;
R ^{2 ′} is H, alkyl, —C (O) NR ⁷ , —C (O) R ^b , cycloalkyl, heterocycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, heteroaryl, or aryl. Yes;

p is 1 to 6;
q is 1 to 6;

R ⁹ is independently H, -CN, -OH, -SH, alkoxy, alkylthio, -CO ₂ R ^{4 '} , -C (O) R ^4a , C (O) NR ⁷ R ⁸ , SO ₂ NR ^4' , NR ^{4 '} R ^5' , SO ₂ -alkyl, -SO ₂ R ^{4 '} , SO ₃ R ^4' , -N = CR ^{4 '} R ^5' , NR ^{4 '} C (O) R ^4'' , -NR ^{4 '-CO- _{haloalkyl, NO 2, -NR 4' -SO}} 2 - haloalkyl, -NR ^{4 '-SO} ₂ - alkyl, NR ^4' CO- alkyl, NR ^{4 '(CH} ₂₎ to _p heteroaryl, alkyl, Hydroxyalkyl, cycloalkyl, halogen, haloalkyl, alkylamino, O (CH ₂ ) _p [O (CH ₂ ) _p ] _q OCH ₃ , -C (NR ^{4 ''} ) NR ^{4 '} benzimidazolyl, C (NR ^4'' ) NR ^4' benzthiazolyl, C (NR ^{4 ''} ) NR ^{4 '} benzoxazolyl, hydroxycycloalkyl, hydroxyalkylamino, haloalkyloxy, heterocycloalkyl, (CH ₂ ) _p NR ⁷ COR ⁸ , aryl, or hetero Represents aryl;

here,
Unless otherwise stated, the C ₁ -C ₆ -alkyl group represents a straight or branched C ₁ -C ₆ -alkyl, preferably a straight or branched chain of 1 to 5 carbon atoms They can be optionally substituted with one or more substituents R ′;
A C ₂ -C ₆ -alkenyl group, unless stated otherwise, represents a straight or branched C ₂ -C ₆ -alkenyl, preferably a straight or branched chain of 2 to 6 carbon atoms. And they can be optionally substituted by one or more substituents R ′;
An alkyl group, unless stated otherwise, is a straight or branched C ₁ -C ₆ -alkyl group, preferably a straight or branched chain of 1 to 6 carbon atoms, straight or branched C Represents a _2- C ₆ alkenyl group, or a linear or branched C ₂ -C ₆ -alkynyl group, which can be optionally substituted by one or more substituents R ′;

R 'is independently H, -CO ₂ R ", -CONHR", -CR "O, -SO ₂ NR", -NR "-CO-haloalkyl, NO ₂ , NR" -SO ₂ -haloalkyl, -NR “—SO ₂ -alkyl, —SO ₂ -alkyl, NR” —CO-alkyl, —CN, alkyl, cycloalkyl, alkylamino, alkoxy, —OH, —SH, alkylthio, hydroxyalkyl, hydroxyalkylamino, halogen, Is haloalkyl, haloalkyloxy, aryl, or heteroaryl;

  R "is independently H, haloalkyl, hydroxyalkyl, alkyl, cycloalkyl, aryl, or heteroaryl;

Cycloalkyl groups represent non-aromatic ring systems containing 3 to 8 carbon atoms, preferably 4 to 8 carbon atoms, wherein one or more carbon atoms in the ring are replaced by the group R ′ as defined above C ₃ -C ₈ -cycloalkyl residues can be -cyclo-C ₃ H ₅ , -cyclo-C ₄ H ₇ , -cyclo-C ₅ H ₉ , -cyclo-C ₆ H ₁₁ , -cyclo May be selected from the group comprising -C ₇ H ₁₃ , -cyclo-C ₈ H ₁₅ ;

  A heterocycloalkyl group represents a non-aromatic ring system comprising 2 to 10 carbon atoms and a heteroatom selected from at least one O, N, and S, wherein one or more carbon atoms in the ring are as defined above. Preferred heterocycloalkyl groups are morpholin-4-yl, piperazinyl, 1-alkylpiperazin-4-yl, piperidinyl, pyrrolidinyl, azepan-1-yl;

  An alkoxy group represents an O-alkyl group, said alkyl group being an alkyl group as defined above; said alkoxy group is preferably a methoxy group, an ethoxy group, an isopropoxy group, a t-butoxy group, or a pentoxy group; A group;

  An alkylthio group represents an S-alkyl group, the alkyl group being an alkyl group as defined above;

A haloalkyl group represents an alkyl group substituted with 1 to 5 halogen atoms, wherein the alkyl group is an alkyl group as defined above; the above-mentioned alkyl group is preferably —C (R ¹⁰ ) ₃ , CR ¹⁰ (R ^{10 '} ) ₂ , -CR ¹⁰ (R ^10' ) R ^{10 ''} , -C ₂ (R ¹⁰ ) ₅ , -CH ₂ C (R ¹⁰ ) ₃ ,, -CH ₂ CR ¹⁰ (R ^{10 ′} ) ₂ , CH ₂ CR ¹⁰ (R ^{10 ′} ) R ^{10 ″} , —C ₃ (R ¹⁰ ) ₇ , or —C ₂ H ₄ C (R ¹⁰ ) ₃ , R ¹⁰ , R ^{10 ′} , R ^{10 ''} represents F, Cl, Br or I, preferably F;

  A hydroxyalkyl group is a HO-alkyl group, which is an alkyl group as defined above;

A haloalkyloxy group represents an alkoxy group substituted with 1 to 5 halogen atoms, said alkyl group being an alkyl group as defined above; said haloalkyloxy is preferably —OC (R ¹⁰ ) ₃ , — OCR ¹⁰ (R ^{10 '} ) ₂ , -OCR ¹⁰ (R ^10' ) R ^{10 ''} , -OC ₂ (R ¹⁰ ) ₅ , -OCH ₂ C (R ¹⁰ ) ₃ , OCH ₂ CR ¹⁰ (R ^{10 '} ) ₂ , -OCH ₂ CR ¹⁰ (R ^{10 '} ) R ^10'' , -OC ₃ (R ¹⁰ ) ₇ , or -OC ₂ H ₄ C (R ¹⁰ ) ₃ and R ¹⁰ , R ^10' , R ^{10 ''} represents F, Cl, Br or I, preferably F;

A hydroxyalkylamino group represents a (HO-alkyl) ₂ -N- group or a HO-alkyl-NH- group, the alkyl group being an alkyl group as defined above;

  An alkylamino group represents an HN-alkyl group or an N-dialkyl group; the alkyl group is an alkyl group as defined above;

  The halogen atom is fluorine, chlorine, bromine or iodine;

An aryl group is an aromatic group having 5 to 15 carbon atoms, which atoms can be substituted with one or more R ′, where R ′ is as defined above; , preferably, a benzyl group, a phenyl _{group, -oC 6 H 4 -R ',} - mC 6 H 4 -R', - pC 6 H 4 -R ', 1- naphthyl, 2-naphthyl, 1-anthracenyl or 2 Anthracenyl, R ′ is as defined above;

  An arylamino group represents an HN-aryl group or an N-diaryl group, said aryl group being an aryl group as defined above;

  A heteroaryl group is a 5- or 6-membered heterocyclic ring system containing at least one heteroatom selected from O, N, and S. The heterocyclic group can be condensed with other aromatic rings. For example, the groups include thiadiazol, thiazol-2-yl, thiazol-4-yl, thiazol-5-yl, isothiazol-3-yl, isothiazol-4-yl, isothiazol-5-yl, oxazole-2 -Yl, oxazol-4-yl, oxazol-5-yl, isoxazol-3-yl, isoxazol-4-yl, isoxazol-5-yl, benzoxazol-2-yl, benzoxazol-4-yl, Benzoxazol-5-yl, benzisoxazol-3-yl, benzisoxazol-4-yl, benzisoxazol-5-yl, 1,2,4-oxadiazol-3-yl, 1,2,4 -Oxadiazol-5-yl, 1,2,5-oxadiazol-3-yl, 1,2,5-oxadiazol-4-yl, 1,2,4-thiadiazol-3-yl, 1 2,4-thiadiazol-5-yl, isothiazol-3-yl, isothiazol-4-yl, iso Azol-5-yl, benzisothiazol-3-yl, benzisothiazol-4-yl, benzisothiazol-5-yl, 1,2,5-thiadiazol-3-yl, 1-imidazolyl, 2-imidazolyl, 1,2,5-thiadiazol-4-yl, 4-imidazolyl, benzimidazol-4-yl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-furanyl, 3-furanyl, 2-thienyl, 3-thienyl 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyranyl, 3-pyranyl, 4-pyranyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl, 2,4-dimethoxy-6-pyrimidinyl , 3-pyridazinyl, 4-pyridazinyl, 2-pyrazinyl, 1-pyrazolyl, 3-pyrazolyl, 4-pyrazolyl, 1,2,3-triazol-4-yl, 1,2,3-triazol-5-yl, 1 , 2,4-triazol-3-yl, 1,2,4-triazol-5-yl, 1,3,5-triazol-6-yl, 2,4-dimethoxy-1,3,5-triazol-6-yl, 1H-tetrazol-2-yl, 1H-tetrazol-3-yl, tetrazolyl, acridyl, furazane, indazolyl, phenazinyl, carbazolyl, phenoxazinyl, indolizine 2-indolyl, 3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl, 1-isoindolyl, 3-isoindolyl, 4-isoindolyl, 5-isoindolyl, 6-isoindolyl, 7-isoindolyl, 2 -Indolinyl, 3-indolinyl, 4-indolinyl, 5-indolinyl, 6-indolinyl, 7-indolinyl, benzo [b] furanyl, benzofurazan, benzothiofurazan, benzotriazol-1-yl, benzotriazol-4-yl, Benzotriazol-5-yl, benzotriazol-6-yl, benzotriazol-7-yl, benzotriazi , Benzo [b] thiophenyl, benzimidazol-2-yl, 1H-benzimidazolyl, benzimidazol-4-yl, benzimidazol-5-yl, benzimidazol-6-yl, benzimidazol-7-yl, benzothiazolyl, quinazolinyl , Quinoxazolinyl, cinnoline, quinolinyl, tetrahydroquinolinyl, isoquinolinyl, tetrahydro-thieno [3,4-d] imidazol-2-one, pyrazolo [5,1-c] [1,2,4] triazine, 2 , 3-Dihydrobenzo [1,4] -dioxin-2-yl, 2,3-dihydrobenzo [1,4] -dioxin-3-yl, 2,3-dihydrobenzo [1,4]- Dioxin-5-yl, 2,3-dihydrobenzo [1,4] -dioxin-6-yl, 2,6-dimethoxypyrimidin-3-yl, 2,6-dimethoxypyrimidin-4-yl, tetrahydroiso Quinolinyl, purine, phthalazine, pteridine, thiatetraazaine Den, thiatriazaindene, isothiazolopyrazine, isothiazolopyrimidine, pyrazolotriazine, pyrazolopyrimidine, imidazopyridazine, imidazo-pyrimidine, imidazopyridine, imidazolotriazine, triazolotriazine, triazolopyridine, triazolopyrazine, tria Zolopyrimidine, 4- [1,2,4] triazolo [4,3-a] pyridin-3-yl, 1-furo [2,3-c] pyridin-4-yl, 1-furo [2,3- c] Pyridin-5-yl, 1-furo [2,3-c] pyridin-3-yl, and triazolopyridazine can be selected. The heterocyclic group may be substituted by one or more R ′, where R ′ is as defined above.

  In the formula (Ih), the following substituents are preferable alone or in combination.

  X is preferably S or O, more preferably S.

R ^3a is preferably H.

In a preferred embodiment, Z is CR ^{2 ′} and R ¹ and R ² together with the C-atom form a 6-membered saturated ring. In another preferred embodiment, Z is N and R ¹ and R ² together with the N atom form a 6 membered saturated ring.

R ⁹ is preferably heteroaryl, aryl or benzyl, more preferably heteroaryl. R ⁹ is more preferably thienopyrimidine, quinazoline, purine, pyrazolopyrimidine or triazolepyrimidine. Even more preferably, R ⁹ is thienopyrimidine.

  t is preferably 0.

  r is preferably 1.

R ^2a is preferably OH, alkyl, aryl or heteroaryl, more preferably alkyl, especially methyl.

  In one preferred embodiment of the present invention, the compound (Ih) in the composition is a compound of the formula (Iha).

Where
R ¹ is COR ^7a ;
R ² and R ^3a are H;
A is NH;
R ⁷ is as defined above;
Z is N;
X is NR ^{2 ′} , O or S;
R ³ is H, methyl, ethyl, methoxy, amine, alkylamine, morpholino, N-methylpiperazine, CF ₃ , or OCF ₃ ;
t is 0;
r is 1; and
R ^2a is optionally substituted aryl, benzyl or heteroaryl.

In another preferred embodiment of the invention, in the compound of formula (Ih), the compound of formula (Iha), wherein R ^7a is optionally substituted aryl, benzyl or heteroaryl.

  Another preferred embodiment of the present invention is a compound of the formula (Ihb) among the compounds of the formula (Ih).

Where
R ^3a is H;
A is NH;
X is NR ^{2 ′} , O or S;
Y ′ is O or NR ^{2 ′} ;
R ^{2 ′} is as defined above;
R ³ is H, methyl, ethyl, methoxy, amine, alkylamine, morpholino, N-methylpiperazine, CF ₃ , or OCF ₃ ;
t is 0;
r is 1; and
R ^2a is an optionally substituted aryl, benzyl, or heteroaryl;
Z is as defined above.

  In the formula (Ihb), the following substituents are preferable alone or in combination.

Z is preferably CR ^{2 ′} .

Y ′ is preferably NR ² ′. In one preferred form, R ^{2 ′} in NR ² ′ of Y ′ is preferably substituted or unsubstituted heteroaryl. In another preferred form, R ^{2 ′ in} NR ² ′ of Y ^′ is aryl. In yet another preferred form, R ^{2 ′ in} NR ² ′ of Y ^′ is benzyl. In a more preferred form, R ^{2 ′} in NR ² ′ of Y ^′ is a pyrimidine or a triazine. In another more preferred embodiment, Y R ² in 'NR ^2' of ^'is substituted heteroaryl or unsubstituted bicyclic, more preferably, be a thienopyrimidine, quinazoline, purine, pyrazolopyrimidine or triazole pyrimidine Even more preferred is thienopyrimidine.

In one preferred form, X is S. In another preferred form, X is O. In yet another preferred form, X is NR ^{2 ′} .

R ³ is preferably H.

R ^2a is preferably aryl or heteroaryl, more preferably phenyl.

  Another preferred embodiment of the present invention is a compound of formula (Ihe) in a compound of formula (Ih).

Where
R ^3a is H; A is NH; X is NR ^{2 ′} , O or S;
R ¹¹ and R ^{12 are} independently H, -CN, -OH, -SH, CO ₂ R ^{4 '} , -C (O) R ^4' , -SO ₂ NR ^{4 '} , NR ^4' R ^{5 '} , SO ₂ -alkyl, -SO ₂ R ^{4 '} , SO ₃ R ^4' , -N = CR ^{4 '} R ^5' , NR ^{4 '} C (O) R ^4'' , -NR ^4' -CO-haloalkyl, NO ₂ , -NR ^{4 '-SO} ₂ - haloalkyl, -NR ^4' -SO ₂ - alkyl, -NR ^{4 '-CO-} alkyl, -NR ^4' (CH ₂₎ to _p heteroaryl, alkyl, cycloalkyl, alkylamino, alkoxy , Alkylthio, O (CH ₂ ) _p [O (CH ₂ ) _p ] _q OCH ₃ , —C (NR ^{4 ″} ) NR ^{4 ′} benzimidazolyl, C (NR ^{4 ″} ) NR ^{4 ′} benzthiazolyl, C (NR ^{4 ''} ) NR ^{4 '} represents benzoxazolyl hydroxyalkyl, hydroxycycloalkyl, hydroxyalkylamino, halogen, haloalkyl, haloalkyloxy, aryl, arylalkyl or heterocycle;
R ^{4 ′} , R ^{4 ″} and R ^{5 ′} are as defined above;
R ^{2 ′} is as defined above;
R ³ is H, methyl, ethyl, methoxy, amine, alkylamine, morpholino, N-methylpiperidine, CF ₃ , or OCF ₃ ;
t is 0;
r is 1; and
R ^2a is optionally substituted aryl, benzyl or heteroaryl.

  In the formula (Ihe), the following substituents are preferred, either alone or in combination.

Z is preferably CR ^{2 ′} .

In one preferred form, X is S. In another preferred form, X is O. In yet another preferred form, X is NR ^{2 ′} .

R ¹¹ is preferably —H, —CN, —OH, halogen, haloalkyl, haloalkyloxy, and more preferably —CN.

R ¹² is preferably H.

R ^2a is preferably aryl or heteroaryl, more preferably phenyl.

  For compounds of formula (Ih), the term “stereoisomer” means cis / trans or E / Z isomerism. More specifically, the possibility that one or more double bonds are present in the various substituents of the compounds of the present invention may be in the E or Z configuration. These pure or non-pure stereoisomers, alone or as a mixture, form an integral part of the compound of formula (Ih). The term “stereoisomer” also refers to isomers that result from the presence of one or more axes and / or centers of symmetry in a molecule, and isomers that cause rotation of a polarized beam, either alone or as a mixture. Including. More specifically, the term includes enantiomers and diastereomers as such or as a mixture.

  The compound of the formula (Ih) used in the present invention may form a salt with an inorganic or organic acid or base. Examples of pharmaceutically acceptable salts are acetates derived from acetic acid, aconite salts derived from aconitic acid, ascorbates derived from ascorbic acid, benzoates derived from benzoic acid, cinnamic acid Derived from cinnamate derived from citric acid, embonate derived from embonic acid, heptanoate derived from heptanoic acid, formate derived from formic acid, derived from glutamic acid Glutamate, glycol salt derived from glycolic acid, chlorine compound derived from hydrochloric acid, bromine compound derived from hydrobromide, lactate derived from lactic acid, maleate derived from maleic acid, Malonate derived from malonic acid, mandelate derived from mandelic acid, methanesulfo derived from methanesulfonate Acid salt, naphthalene-2-sulfonate derived from naphthalene-2-sulfonic acid, nitrate derived from nitric acid, perchlorate derived from perchloric acid, phosphate derived from phosphoric acid, From phthalates derived from phthalic acid, salicylates derived from salicylic acid, sorbic acids derived from sorbic acid, stearates derived from stearic acid, succinates derived from succinic acid, sulfuric acid Non-toxic inorganic or organic salts such as sulfates derived, tartrate derived from tartaric acid, p-toluenesulfonate derived from p-toluenesulfonic acid, and others are included without limitation. Such salts can be prepared by methods known to those skilled in the art and by methods described in the prior art.

  Other salts that are not considered pharmaceutically acceptable salts, such as oxalates derived from oxalic acid, are compounds of formula (Ih), pharmaceutically acceptable salts thereof, pharmaceutically acceptable In the production of prodrugs or stereoisomers thereof, they may be suitable as intermediates.

  The present invention covers pharmaceutically acceptable salts as indicated above, but also allows for suitable separation or crystallization of the compound of formula (Ih), for example chiral amines. It also extends to salts such as the salt obtained together.

  The compounds of the above formula (Ih) include prodrugs of these compounds. As used herein, the term “prodrug”, although administered to a patient once per se (“prodrug”), is not pharmaceutically active, but chemically and / or biologically those drugs. In a biologically active form (compound of formula (Ih)), i.e. a compound that is converted when the compound is administered. Prodrugs include, for example, compounds in which the hydroxyl group, amine group, or sulfhydryl group of the compound of the present invention is bonded to any group and cleaved when administered to a patient to form a hydroxy group, amine group, or sulfhydryl group. It is. Thus, representative examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of alcohol, sulfhydryl and amine functional groups of the compounds of the present invention. Further, in the case of carboxylic acid (—COOH), esters such as methyl ester, ethyl ester, and 2-ester may be used. Esters may themselves be active and / or may be hydrolysable under in vivo conditions in the human body.

-Cytokine according to the present invention Cytokines are proteins secreted from cells that mediate or regulate immunity, inflammation and hematopoiesis. They must be generated de novo in response to immune stimulation. Responses to cytokines include increasing or decreasing the expression of membrane proteins (including cytokine receptors) and secretion and proliferation of effect molecules.

  The composition of the invention comprises a cytokine selected from the group of (a) autocrine cytokines, (b) paracrine cytokines, (c) endocrine cytokines. Autocrine cytokines act on the cells that secrete them. The action of paracrine cytokines is limited to the vicinity of cytokine secretion, and endocrine cytokines spread to distant parts of the body. The composition may also include one or more cytokines.

  The composition of the present invention comprises (i) four α-helix cytokine families: (a) an IL-2 subfamily, (b) an interferon (IFN) subfamily, and (c) an IL-10 subfamily. A cytokine family consisting of a family (including several non-immunological cytokines such as erythropoietin (EPO) and thrombopoietin (THPO)), (ii) the IL-1 family (eg IL-1 and IL-18), (iii Preferably, it comprises a cytokine selected from the group of)) the IL-17 family, and (iv) the cytokine family of chemokines.

  The IL-1 family is preferred. Interleukin-2 belongs to the family of cytokines and includes IL-4, IL-7, IL-9, IL-15 and IL-21. IL-2 is a IL-2 receptor α (CD25) specific for IL-2, IL-2 receptor β (CD122), and a common γ chain (shared by all members of this cytokine family ( Signals are sent through a receptor complex consisting of γc). IL-2 binding activates Ras / MAPK, JAK / Stat and PI 3-kinase / Akt signaling modules.

  The interferon family is more preferred. The family includes type I interferons, type II interferons, and type III interferons. Type I interferons are IFN-α (alpha), IFN-β (beta), IFN-κ (kappa), IFN-δ (delta), IFN-ε (epsilon), IFN-τ (tau), IFN-ω (Omega) and IFN-ζ (also known as zeta, limitin). Type II contains the only member IFN-γ (gamma). Type III interferons include IFN-λ (lambda) molecules, referred to as IFN-λ1, IFN-λ2, and IFN-λ3 (also referred to as IL29, IL28A, and IL28B, respectively). Types I and II are preferred. More preferred are IFN-α (alpha), IFN-β (beta) and IFN-γ (gamma).

  Tumor necrosis factor (TNF) is more preferred. Here, this includes TNF-α (alpha) and TNF-β (beta).

・ IFN-α
In a particularly preferred embodiment, IFN-α is a cytokine in the composition. The 13 human IFN-α gene encodes 12 different proteins. The IFN-α proteins share 76-99% sequence homology and their biological activity differences are slight, so the commercial TFN-α is TFN-α1. IFN-α administered subcutaneously showed an unexpected broad spectrum of efficacy in hematological malignancies including putative B-cell, T-cell and myeloid lineage tumors. In several diseases such as hairy cell leukemia and chronic myeloid leukemia, IFN-α has been widely effective. However, IFN-α is currently less effective in treating hairy cell leukemia, plays a limited role, and is for selected patients who have failed nucleoside treatment.

  Commercially, the most important use of INF-α is for the treatment of hepatitis C (HCV). HCV infection is widespread throughout the world. WHO estimates that approximately 3% of the world's population is HCV carriers. The genetic heterogeneity of HCV among the virus quasispecies within an individual patient, plus the considerable antigenic variation, probably explains the difficulty of vaccine development.

  IFN-α treatment of chronic hepatitis C patients has only achieved a sustained antiviral response in 15-20% of patients. Combination treatment with ribavirin resulted in a sustained response of 38-43%. Additional improvements in HCV treatment were achieved with the development of peg IFN-α to improve the pharmacokinetic profile. The response rate of peg IFN-α is within the scope of IFN-α ribavirin combination therapy.

  Current treatment is peg IFN-α and ribavirin. The brand names are Copegus (Hoffman la Roche) and Rebetol (Schering-Plow). This treatment resulted in an average response of 50-60%. HCV gene type 1 has a response rate of only 42-45%.

  Due to the high proportion of non-responders to standard treatment, improvements in HCV treatment are still significantly needed

  In one preferred embodiment, the invention provides a composition of the invention wherein the cytokine is IFN-α and the compound is a compound of general formula (Ih), HCV treatment, early and late chronic hepatitis C, and Chronic hepatitis B, chronic hepatitis D, bunyamwera virus, positive-sense ssRNA virus, picornavirus, flavivirus, coronavirus, influenza virus, avian influenza virus, human solid and hematological malignancies, hairy Cellular leukemia, renal cancer, basal cell carcinoma, malignant myeloma, Kaposi's sarcoma, multiple myeloma, chronic and malignant leukemia, non-Hodgkins lymphoma, Burkitt lymphoma, follicular lymphoma, cryptocox meningitis, true red blood cell Augmentation, essential thrombocythemia, uveitis, laryngeal papillomatosis, mycosis fungoides, warts, warts, multiple sclerosis, chronic granulomatosis, From the group of viral infections such as psoriasis, infections, mycobacterial infections, leprosy, Mycobacterium tuberculosis infection, tuberculosis, leishmaniasis, HIV opportunistic infections, stroke, myocardial ischemia It relates to the use for the manufacture of a medicament for the treatment of selected diseases.

  The following indications are preferred: chronic hepatitis C, chronic hepatitis B, chronic hepatitis D, influenza infection, HIV infection, stroke, myocardial ischemia, multiple sclerosis, osteoporosis.

  The following indication is particularly preferred: chronic hepatitis C.

・ IFN-β
IFN-β is preferred in the composition. The use of IFN-γ significantly limits multiple sclerosis (MS). MS affects approximately 2.5 million people worldwide. MS is twice as popular among women as men. IFN-β is an anti-inflammatory and immunoregulatory interferon that reduces the frequency of multiple sclerosis (MS) recurrence and ameliorates experimental autoimmune neuritis.

  As a result, human recombinant interferon-beta has been approved by the FDA for patients with relapsed MS, and its safety profile is well characterized.

  In one preferred embodiment, the present invention provides a composition of the present invention wherein the cytokine is IFN-β and the compound is a compound of the general formula (Ih), chronic hepatitis C, chronic hepatitis B, chronic D Hepatitis, bunyamwera virus, positive-sense ssRNA virus, picornavirus, flavivirus, coronavirus, influenza virus, avian influenza virus, human solid and hematological malignancies, hairy cell leukemia, renal cancer, basal cell Cancer, malignant myeloma, Kaposi's sarcoma, multiple myeloma, chronic and malignant leukemia, non-Hodgkins lymphoma, Burkitt lymphoma, follicular lymphoma, cryptococcal meningitis, polycythemia vera, essential thrombocythemia , Uveitis, laryngeal papillomatosis, mycosis fungoides, warts, multiple sclerosis, chronic granulomatosis, osteoporosis, infection, myco Treatment of diseases selected from the group of viral infections such as Kuteria infection, leprosy, M. tuberculosis infection, tuberculosis, leishmaniasis, human opportunistic infection in HIV infection, stroke, myocardial ischemia For the manufacture of a medicament for use.

  The following indications are preferred: chronic hepatitis C, chronic hepatitis B, chronic hepatitis D, influenza infection, HIV infection, stroke, myocardial ischemia, multiple sclerosis, osteoporosis.

  The following indication is particularly preferred: multiple sclerosis

・ IFN-γ
Human IFN-γ is a preferred cytokine in the composition. This is a glycated protein and has slight sequence homology with IFN-α and IFN-β. IFN-γ is produced by activated T cells and natural killer cells in response to specific growth stimuli. While IFN-γ shares many activities with Type I interferons, it has many other activities that result in its original role being the regulation of the immune system. The clinical application of IFN-γ is limited to rare diseases. IFN-γ is FDA approved for chronic granulomatosis (CGD). CDG is a rare congenital anomaly that occurs in patients, primarily children, and is caused by the vulnerability of severe recurrent bacterial and novel infections.

  The second FDA approved clinical application is for congenial osteoporosis, a life-threatening pediatric disease. The most common adverse events during IFN-γ treatment are fever, headache, nausea and vomiting.

  In one preferred embodiment, the present invention provides a composition of the present invention wherein the cytokine is IFN-γ and the compound is a compound of the general formula (Ih), chronic hepatitis C, chronic hepatitis B, chronic D Hepatitis, bunyamwera virus, positive-sense ssRNA virus, picornavirus, flavivirus, coronavirus, influenza virus, avian influenza virus, human solid and hematological malignancies, hairy cell leukemia, renal cancer, basal cell Cancer, malignant myeloma, Kaposi's sarcoma, multiple myeloma, chronic and malignant leukemia, non-Hodgkins lymphoma, Burkitt lymphoma, follicular lymphoma, cryptococcal meningitis, polycythemia vera, essential thrombocythemia , Uveitis, laryngeal papillomatosis, mycosis fungoides, warts, multiple sclerosis, chronic granulomatosis, osteoporosis, infection, myco Treatment of diseases selected from the group of viral infections such as Kuteria infection, leprosy, M. tuberculosis infection, tuberculosis, leishmaniasis, human opportunistic infection in HIV infection, stroke, myocardial ischemia For the manufacture of a medicament for use.

  The following indications are preferred: chronic hepatitis C, chronic hepatitis B, chronic hepatitis D, influenza infection, HIV infection, stroke, myocardial ischemia, multiple sclerosis, osteoporosis, CDG.

  The following indications are particularly preferred: CDG, congenial osteoporosis.

・ TNF
Tumor necrosis factor alpha (TNF) is a preferred cytokine in the composition, which is a multifunctional cytokine that was isolated 30 years ago and plays an important role in apoptosis and cell survival as well as inflammation and immunity . Although named for its antitumor properties, TNF has been implicated in a wide range of other diseases. In cancer, TNF is currently used in the local therapy of partially advanced soft tissue sarcomas and metastatic melanomas, and tumors of any other unresectable tissue to avoid amputation of the limbs . It has been shown that TNF acts synergistically with cytostatics in isolated limb perfusion. High amounts of TNF-α can be safely and locally administered through isolated perfusion of the affected limb, with additional chemotherapy, with or without IFN-γ. This treatment resulted in a complete remission of between 70% and 80% in the case of melanoma metastases, and a complete remission of between 25% and 36% in the case of non-eradicable soft tissue sarcomas. . Including dual targeting; TNF-α and IFN-g cause apoptosis of vascular cells, whereas melphalan causes apoptosis of tumor cells (Curr Opin Immunol1998 Oct; 10 (5): 573- 80).

  In one preferred embodiment, the present invention provides a composition for treatment of a disease selected from the group of solid cancers and hematopoietic cancers, wherein the cytokine is TNF and the compound is a compound of general formula (Ih). For the manufacture of a medicament for the use.

  The following indication is preferred: solid cancer.

  The following indications are particularly preferred: soft tissue sarcoma and metastatic melanoma

・ IL-2
IL-2 is a preferred cytokine in the composition. Human immunodeficiency virus type 1 (HIV-1) gene expression and transcription are essential steps in the viral life cycle and are potential targets for the suppression of HIV-1 replication. Infection with HIV-1 results in progressive death of CD4 T-cells, resulting in the development of AIDS. The mechanism by which this death of CD4 T-cells is still not fully understood, but the death of CD4 T-cells caused by HIV-1 is now recognized as the center of immune deficiency. Antiretroviral therapy (ART) combined with IL-2 substantially increased CD4 cells and decreased viral load more dramatically than ART alone (Davey RT et al., JAMA. 2000 Oct 25; 284 (16): 2055-6).

  In one preferred form, the invention selects a composition of the invention wherein the cytokine is IL-2 and the compound is a compound of general formula (Ih) from the group of viral infections, in particular HIV infections It relates to the use for the manufacture of a medicament for the treatment of diseases.

  In one embodiment of the invention, the active substances according to the invention, ie the compound of general formula (Ih) and the cytokine are not present in one composition, but are administered separately but simultaneously. As used herein, the term “simultaneously” means that (i) a plurality of substances are administered separately and completely simultaneously, or (ii) one or another substance is administered first, and a second substance is It is meant to be administered immediately thereafter, but at a time when the additional and / or synergistic effects shown here are maintained. In one preferred embodiment of the foregoing description, for example, cytokines are administered by injection, while the compound of general formula (Ih) is separately and simultaneously, but simultaneously (wherein In the meaning outlined in).

  In the present invention, “treatment” is intended to mean to completely or partially heal the disease, or to alleviate the disease or to stop the progression of a given disease.

  Thus, in one aspect, the invention provides a composition of the invention, i.e., a cytokine and a compound of formula (Ih), a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable prodrug, or a steric form thereof. It relates to the use of a composition comprising isomers, optionally containing suitable auxiliaries and additives, for the manufacture of a medicament.

  The compound of formula (Ih) may be present in the composition as a pharmaceutically acceptable salt, a pharmaceutically acceptable prodrug, or a stereoisomer thereof, and if desired, suitable adjuvants And may be present in the composition along with additives.

  Further, the invention includes cytokines, compounds of formula (Ih), pharmaceutically acceptable salts thereof, pharmaceutically acceptable prodrugs, or stereoisomers thereof, and cytokines as outlined above. The present invention relates to a method for treating or preventing a disease, comprising administering an effective amount of the composition.

  A composition comprising a cytokine and a compound of formula (Ih) in free or pharmaceutically acceptable salt form and pharmaceutically acceptable prodrug form is a pharmaceutically acceptable diluent thereof Or a carrier may be included.

  Compositions of the present invention containing cytokines and compounds of general formula (Ih), pharmaceutically acceptable salts thereof, are used as treatments in animals, preferably mammals, and especially humans, dogs and chickens. In addition to conventional or pharmaceutically harmless excipients and additives, as a mixture with others or as pharmaceuticals, i.e. enteral or parenteral use, in addition to the compounds of formula (Ih) or their It can be administered in the form of a medicament comprising an effective amount of at least one salt. Said composition comprising a cytokine and a compound of formula (Ih) can also be administered in the form of their salts obtained by reacting physiologically acceptable acids and bases with the respective compounds.

  Production of the medicament containing the composition of the present invention and its application can be carried out according to well-known dispensing methods.

A composition of the invention comprising a cytokine and a compound of formula (Ih) may be administered in the form of a crude compound when used for therapy, preferably introducing an active ingredient, optionally,
A physiologically acceptable salt form in a pharmaceutical composition comprising one or more adjuvants, excipients, carriers, buffers, diluents, and / or other conventional pharmaceutical auxiliaries. And is preferably administered. The salts of such compounds may be anhydrides or solvated.

  The carrier or carriers must be acceptable in the sense of being compatible with the other ingredients of the composition and not deleterious to the recipients thereof.

  The agents of the present invention can be oral, rectal, bronchial, nasal, topical absorption, oral, sublingual, transdermal, intravaginal, or parenteral (skin, subcutaneous, intramuscular, (Including intraperitoneal, intravenous, intraarterial, intracerebral, intraocular injection or infusion), and administration by inhalation or insufflation containing powders and liquid aerosols, or by sustained release formulations It may be an embodiment suitable for the above. Suitable examples of sustained release formulations include a hydrophobic solid polymer semi-permeable matrix comprising a compound of the invention, which matrix may be in the form of a molded article such as a film or microcapsule, for example.

  The compositions of the present invention may be administered in dosage units in the form of a medicament, together with conventional adjuvants, carriers, or diluents. Such forms include solids, specifically tablets, filled capsules, powders and pills, etc., and liquids, specifically aqueous or non-aqueous solutions, suspensions, emulsions, especially And tablets, elixirs, and capsules filled therewith, all used as injectable solutions sterilized for oral, rectal administration, and parenteral use It is. Such agents and dosage unit forms thereof may contain conventional ingredients in conventional proportions, with or without additional active compounds and major components, and such dosages. The unit form may contain any effective amount of the active ingredient commensurate with the intended daily dosage range to be used.

  The compositions that can be used in the present invention are administered in a wide variety of oral and parenteral dosage forms.

  For preparing a medicament from the composition of the invention, the composition comprising a cytokine and a compound of formula (Ih) or a pharmaceutically acceptable carrier thereof may be solid or liquid. Solid form preparations include powders, tablets, pills, capsules, suppositories, and dispersible granules. A solid carrier contains one or more substances that may act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, preservatives, disintegrating tablets, or an encapsulating material. Can do.

  In powders, the carrier is a finely divided solid in a mixture with the finely divided active compound. In tablets, the active ingredient is mixed with the carrier having the necessary and suitable proportions of binding capacity and compressed to the desired shape and size. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term “formulation” refers to an agent of an active compound with an encapsulating material providing a capsule as a carrier in which the active compound is surrounded by and thus associated with the carrier, with or without the carrier Includes type. Similarly, capsules and medicinal candies are included. Tablets, powders, capsules, pills, cachets, and medicinal candies can be used as solid forms suitable for oral administration.

  For preparing suppositories, a low melting wax such as fatty acid glyceride or cocoa butter is first melted and the active component is dispersed homogeneously therein by stirring. The molten homogeneous mixture is then poured into convenient sized molds, cooled and thereby solidified. Compositions suitable for vaginal administration may be provided as pessaries, tampons, creams, gels, pastes or sprays containing active ingredients such as carriers known to be suitable in the art. Liquid formulations include solutions, suspensions, and emulsions, for example, solutions such as water or water-propylene glycol solutions, suspensions, and emulsions. For example, liquid preparations for injection can be prepared as solutions in aqueous polyethylene glycol solution.

  Said composition comprising a cytokine and a compound of formula (Ih) according to the present invention may be prepared for parenteral administration (eg administration by injection such as intravenous bolus or continuous infusion) or ampoules pre-filled It may be provided in the form of a dosage unit such as a syringe, a small amount of infusion, or in multiple dosage forms with added preservatives. The composition may take the form of a suspension, solution, or emulsion in an oily or aqueous vehicle, and formulation aids such as suspending, stabilizing and / or dispersing agents. May be included. Alternatively, the active ingredient may be in the form of a powder by aseptic separation of a sterile solid or by lyophilizing a solution in which a suitable vehicle, such as sterile, pyrogen-free water, is formed prior to use. Also good.

  Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizers, and thickening agents as desired. Aqueous suspensions suitable for oral use are finely divided into water, with viscous materials such as natural or synthetic thickeners, resins, methylcellulose, sodium carboxymethylcellulose, or other known suspending agents. It can be prepared by dispersing the active ingredient.

  Also included are solid formulations that are converted to liquid formulations for oral administration immediately prior to use. Such liquid forms include solutions, suspensions, and emulsions. These preparations may contain, in addition to the active ingredient, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, and the like.

  In one form of the invention, the medicament is given locally, systemically or by a combination of the two routes.

  Preferably, the medicament is an ointment, gel, salve, emulsion, lotion, foam, mixed phase or amphiphilic emulsifying (oil / water-water / oil mixed phase) cream, liposome, transfersome. ), Prepared in the form of a paste or powder.

  Ointments and creams may be prepared, for example, using aqueous or oily bases with suitable thickening and / or gelling agents added. Lotions may be prepared using an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents, or coloring agents. I will.

Compositions suitable for topical administration in the mouth include medicinal candy flavored with the active agent, generally in a substrate such as sucrose and acacia or tragacanth; the active agent in gelatin, glycerin Or a troche containing in an inert substrate such as sucrose and acacia or tragacanth; and a mouthwash containing the active ingredient in a suitable liquid carrier.

  Solutions or suspensions are applied directly into the nasal cavity by conventional means, for example with a dropper, pipette or spray. The composition may be provided in a single dose or multiple dose form. In the case of the latter dropper or pipette, the patient may be achieved by administering a predetermined amount of a solution or suspension in a predetermined amount. In the case of spraying, it may be achieved, for example, by means of a metering pump.

  Also, for administration to the respiratory tract, the active ingredient is in combination with a suitable propellant such as chlorofluorocarbon (CFC) such as dichlorodifluoromethane, trichlorofluoromethane or dichlorotetrafluoroethane, carbon dioxide, or other suitable gas, It may be achieved by means of an aerosol dosage form that is in a compressed state. The aerosol may conveniently contain a surfactant such as lecithin. The amount of drug may be adjusted according to the conditions of the measurement valve.

  Alternatively, the active ingredient is in the form of a dry powder, such as a mixed powder in which the compound is mixed in a suitable powder base such as lactose, starch, hydroxypropylmethylcellulose and polyvinylpyrrolidone (PVP) and other starch derivatives. May be provided in Conveniently, the powder carrier will form a gel in the nasal cavity. The powder composition may be provided, for example, in dosage unit form, in a blister pack in which the composition is administered in capsules or drug sachets such as gelatin, or by means of an inhaler.

  Compositions for administration into the respiratory tract include nasal administration compositions in which the compound generally has a small particle size, for example of the order of 5 microns or less. Will. Such a particle size may be obtained, for example, by means known in the art, for example by micronization.

  If desired, compositions compatible with sustained release of the active ingredient may be used.

  The pharmaceutical dosage form is preferably in the form of a dosage unit. In such form, the dosage form is subdivided into dosage units containing appropriate quantities of the active ingredient. The dosage unit form may be a packaged dosage form, a packaged tablet, a package containing a separate dosage form such as a capsule, and a powder in a vial or ampoule. Also, the dosage unit may be a capsule, a tablet, a capsule, a medicinal candy itself, or some suitable number of these may be packaged. Tablets or capsules for oral administration, liquids for intravenous injection and continuous infusion are preferred compositions.

  In one preferred form, the composition is prepared according to conventional procedures as a pharmaceutical composition adapted for intravenous administration to humans. Typically, compositions for intravenous administration are sterile isotonic aqueous buffer solutions. If necessary, the composition may include a solubilizing agent and a local anesthetic such as lidocaline to ease pain at the site of injection. In general, the ingredients are either separately or mixed in the form of a dosage unit, such as a lyophilized powder or a water-free concentrate, in a sealed container such as an ampoule or sachet indicating the amount of active agent. Supplied. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. Where the composition is administered by injection, an ampoule of sterile saline can be provided so that the ingredients may be mixed prior to administration.

  The present invention provides a pharmaceutical pack or kit comprising one or more containers filled with one or more components of the pharmaceutical composition of the present invention. Optionally, with such one or more containers, a notice may be made by an administrative body that regulates the manufacture, use, or sale of a pharmaceutical or biological product, and the notice may be posted to humans. Indicates that the manufacture, use or sale has been approved by the institution for administration. Thus, the present invention also relates to a pack or kit comprising therein a cytokine and the compound of general formula (Ih) separately.

  In other embodiments, for example, the cytokine in the composition may be administered in a controlled release system. For example, the polypeptide is administered using intravenous infusion, implantable osmotic pumps, transdermal patches, liposomes, or other modes of administration. In one embodiment, a pump will be used (Sefton (1987) CRC Crit. Ref. Biomed. Eng. 14: 201; Buchwald et al. (1980) Surgery 88: 507; Saudek et al. (1989) N Eng. J. Med. 321: 574). In other embodiments, the compound (Ih) can be administered in a vesicle state, particularly as a liposome (Langer (1990) Science 249: 1527-1533; Treat et al. (1989) in Liposomes in the Therapy of (See Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, NY, 353-365; WO 91/04014; US 4,704,355.) In other embodiments, polymeric materials may be used (Medical Applications of Controlled Release (1974) Langer and Wise (eds.), CRC Press: Boca Raton, Fla .; Controlled Drug Bioavailability, Drug Product Design and Performance, (1984) Smolen and Ball (eds.), Wiley: NY; Ranger and Peppas (1953) J. Macromol. Sci. Rev. Macromol. Chem. 23: 61; see also Levy et al. (1985) Science 228: 190; During et al. (1989) Ann. Neurol. 25: 351; (See Howard et al. (1989) J. Neurosurg. 71: 105).

  In yet another aspect, the controlled release system is placed near the therapeutic target, i.e., the target cell, tissue or organ, thus requiring very little of the systemic dose (e.g., Goodson (1984 ) 115-138 in Medical Applications of Controlled Release, vol. 2), other controlled release systems are discussed in a review by Langer (1990, Science 249: 1527-1533).

  In certain embodiments, it is preferred to administer the pharmaceutical composition of the present invention locally to the area in need of treatment; for example, in combination with local injection during surgery, for example post-surgical wound dressings Topical application, injection, catheter means, suppository means, or implant means (wherein the implant is a porous, non-porous, or gel-like material comprising a membrane, such as a silastic membrane, or fiber) However, the present invention is not limited to these.

  More details on dosage forms and administration techniques may be found in the latest edition of Remington's Pharmaceutical Sciences (Maack Publishing Co. Easton, Pa.).

  The pharmaceutical composition may comprise two or more of the compounds of formula (Ih) or a pharmaceutically acceptable salt thereof, and further other therapeutically active substances. The pharmaceutical composition can also include two or more cytokines, and still other therapeutically active substances.

  A suitable amount of compound (Ih) administered to a human will be in the range of 5 to 500 mg.

  To prepare the pharmaceutical dosage form, pharmaceutically inert inorganic or organic excipients can be used. For preparing pills, tablets, coated tablets and hard gelatin capsules, for example, lactose, corn starch or derivatives thereof, talc, stearic acid or salts thereof and the like can be used. Excipients for soft gelatin capsules and suppositories are, for example, fats, waxes, semi-solid and liquid polyols, natural or solidified oils and the like. Suitable excipients in solution and syrup products are, for example, water, sucrose, invert sugar, glucose, polyol and the like. Suitable excipients in injectable solution products are, for example, water, alcohols, glycerol, polyols or vegetable oils.

  The amount of formula (Ih) in the composition can be varied within wide limits and is adapted to the individual conditions in each individual case. For the above uses, the dosage will vary depending on the mode of administration, the particular condition being treated and the effect desired. In general, however, satisfactory results are achieved at a weight ratio of about 1 to 100 mg / kg animal weight, preferably 1 to 50 mg / kg animal weight. In general, suitable dose ratios for larger animals, such as humans, are conveniently about 10 mg to 3 g in a single dose, in 2-4 divided doses per day, or in sustained release form. / Day order.

  For compound (Ih) in the composition, a daily dose of about 10 mg to 500 mg, preferably 50 to 500 mg per person is suitable for oral administration. For other forms of administration, the daily dose is in a similar range. In topical administration, depending on the skin penetration, the type and severity of the disease, and depending on the type and frequency of the dosage form, the concentration of the active compound in the drug that produces a therapeutic effect by topical administration varies. There is a case. Preferably, the concentration of the active compound, pharmaceutically acceptable salt thereof, physiologically functional derivative thereof or stereoisomer thereof in the medicament according to the invention is between 1 μmol / l and 100 mmol / l. is there.

  The following examples and drawings are included to demonstrate preferred forms of the invention. In order for the techniques disclosed in the examples to function well in the practice of the present invention, the techniques discovered by the inventor are followed and they can be recognized as preferred embodiments for the practice. Will be understood by those skilled in the art. However, one of ordinary skill in the art appreciates that the present disclosure can be modified as appropriate to the disclosed aspects without departing from the scope and spirit of the invention as set forth in the appended claims in light of the present disclosure. It is. All references cited are incorporated herein by reference.

  Abbreviations: min, minutes; h, hours; r.t., room temperature; t-, tert-.

NMR spectra: Bruker Avance 300 MHz. Spectra show residual solvent peaks (DMSO-d ₆ , ( _H = 2.49; CD ₃ OD, ( _H = 3.31; CDCl ₃ , ( _H = 7.26; CD ₃ CN, ( _H = 1.93; (CD ₃ ) ₂ CO, ( _H = 2.05), respectively, recorded at 300 MHz ( ¹ H-NMR).
Analytical LC / ESI-MS: 2 x Waters 600 multisolvent delivery system. 50 μl sample loop. Column, Chromolith Speed ROD RP18e (Merck, Darmstadt), 50 x 4.6 mm, 2 μm pre-filter (Merck). Eluent A, H ₂ O + 0.1% HCO ₂ H; Eluent B, MeCN. Gradient, 5% B to 100% B within 5 minutes; flow rate, 3 ml / min. Waters LCZ single quadrupol Mass spectrometer with attached electric spray light source. MS method, MS8minPM-80-800-20V; positive / negative ion mode scanning, m / z 80-800 in 1 second; capillary, 3.5 kV; cone voltage, 20 V; multiplication voltage, 400 V; probe and solvent removal Japanese gas temperature, 120 ° C and 350 ° C, respectively. Waters 2487 dual λ absorbance detector, set to 254 nm.

Preparation HPLC-MS: Waters 600 Multisolvent Delivery System. 2000 μl or 5000 μl sample loop. Column, Waters X-Terra RP18, 7 μm, 19 x 150 mm included X-Terra RP18 protection cartridge 7 μm, 19 x 10 mm; use flow rate 20 ml / min, or YMC ODS-A, 120 A, 40 x X-Terra RP18 protection cartridge with 150 mm 7 μm, 19 x 10 mm; flow rate when used 50 ml / min. Preparation _{solvent: MeCN-H 2 O -HCO 2} H 80: 20: 0.05 (v: v: v). Eluent A, H ₂ O + 0.1% HCO ₂ H; Eluent B, MeCN. Different linear gradients 5-100% eluent B, adapted to the sample. Injection volume: 500 μl-2000 μl depending on the sample. Waters ZQ single quadrupol mass spectrometer attached electric spray light source. Positive / negative ion mode scanning m / z 80-800 per second; capillary, 3.5 kV or 3.0 kV; cone voltage, 20 V; multiplication voltage, 400 V; probe and desolvation gas temperature, 120 ° C and 350 respectively ° C. Mass-triggered fraction collection attached to Waters Fraction Collector II. Waters 996 Photodiode array detector.

Synthesis of (Ih) The compound of formula (Ih) may be obtained in various ways.
Piperidin-4yl-thiazole-4-carboxamide can be prepared by various methods described in the literature. An example is the oxidation of suitable 2,5-dihydrothiazoles, as described in Houben-Weyl, 2002, 730. Dihydrothiazoles can also be found in the above literature, or You, S., Razavi, H., Kelly, JW Angew. Chem. 2003, 115, 87, or Katritzky, AR., Cai, C., Suzuki, K., Singh, SK. J. Org. Chem. 2004, 69, 811-814, and their references. An alternative method is described in Yasuchika, S. et. Al. Heterocycles, Vol. 57, No. 5, 2002.

  The compound of the present invention having a piperidin-4-yl substituent at the 2-position of the thiazole ring can be synthesized, for example, according to the following scheme. This synthetic route is partly described in WO2004 / 058750.

2- (1- (tert-butoxycarbonyl) piperidin-4-yl) thiazole-4-carboxylic acid can be converted to the appropriate R ¹ amide by coupling with HBTU, DIPEA in DMF. Different R ¹ -amines are commercially available or can be readily synthesized. The Boc-protecting group can be removed under standard conditions, such as 2 to 3 hours at 0 ° with TFA or 2 to 3 hours with 4N HCl in dioxane. The deprotected piperidino derivative hydrochloride can then be converted to the corresponding amides, ureas, and N-heterocyclic analogs as follows.

  Urea-substituted piperidino compounds can be synthesized by coupling with commercially available isocyanates in the presence of DIPEA. Piperidino compounds substituted with heterocycles can be synthesized by standard means, such as coupling with the corresponding chloroheterocycle in the presence of a base. Alternatively, piperidino compounds substituted with heterocycles can also be obtained by palladium-mediated cross coupling. As another alternative, the hydroxypyridine derivative can be coupled with the piperidino compound by the HBTU coupling method.

  One synthetic route for compounds of the present invention in which piperidin-4-yl is substituted at the 2-position of the thiazole ring is shown in the following scheme.

  The Boc-protecting group can be removed under standard conditions, such as 2 to 3 hours at 0 ° with TFA or 2 to 3 hours with 4N HCl in dioxane. The deprotected piperidino derivative hydrochloride can then be converted to the corresponding N-heterocyclic analog in various ways as described above.

Synthesis of 4- (methoxy-methyl-carbamoyl) -piperidine-1-carboxylic acid t-butyl ester Piperidine-1,4-dicarboxylic acid mono-tert-butyl ester (1.0 eq, 21.8 mmol) in 35 ml dry N, N -Dissolved in dimethylformamide under inert conditions. O, N-dimethyl-hydroxylamine hydrochloride (1.03 eq, 22.5 mmol), benzotriazol-1-ol monohydrate (1.03 eq, 22.5 mmol) and triethylamine (1.5 eq, 32.7 mmol) were added. The reaction mixture is cooled to 0 ° C., N- (3-dimethylaminopropyl) -N-ethylcarbodiimide hydrochloride (1.0 eq, 21.8 mmol) is added over 10 minutes, and the mixture is added at 0 ° C. for 1 hour and room temperature. And stirred vigorously for 18 hours.

The solvent was removed under vacuum and the residue was suspended in ethyl acetate. The organic layer was extracted three times with 100 ml of 1M citric acid, twice with aqueous sodium carbonate and 100 ml brine, dried over MgSO ₄ and filtered. The solvent was removed and the residue was purified by distillation to give a yield of 80%.

・ Synthesis of 4-formyl-piperidine-1-carboxylic acid t-butyl ester
4- (Methoxy-methyl-carbamoyl) -piperidine-1-carboxylic acid tert-butyl ester (1.0 eq, 16.4 mmol) was dissolved in 100 ml dry tetrahydrofurane under an inert atmosphere. The solution was added dropwise at -50 ° C. over 1 hour into a suspension of lithium alanate (3.0 eq, 49.6 mmol) suspended in 70 ml of dry tetrahydrofuran. During the addition of the mixture, the temperature was maintained at −50 ° C. and warmed to 0 ° C. within 3 hours.

The mixture was cooled to −78 ° C. and carefully quenched with 100 ml of 1M citric acid. The mixture was warmed to room temperature and diluted with 400 ml of ethyl acetate. The layers were separated and the aqueous layer was extracted 3 times with 70 ml of ethyl acetate. The organic layer was extracted three times with 100 ml of 1M citric acid, twice with aqueous sodium carbonate and 100 ml brine, dried over MgSO ₄ and filtered. The solvent was removed and the residue was purified by distillation to give a yield of 85%.

Synthesis of 4- (4-ethoxycarbonyl-4,5-dihydro-thiazol-2-yl) -piperidine-1-carboxylic acid t-butyl ester
4-Formyl-piperidine-1-carboxylic acid tert-butyl ester (1.0 eq, 13 mmol) was dissolved in 40 ml of toluene under inert conditions. To this solution was added L-cysteine ethyl ester hydrochloride (1.6 eq, 21 mmol) and triethylamine (1.6 eq, 21 mmol). The mixture was refluxed for 14 hours. The resulting water was removed with a Dean & Stark trap.

The solvent was removed and the residue was dissolved in 100 ml ethyl acetate. The organic layer was extracted three times with 50 ml of 1M citric acid, twice with aqueous potassium hydrogen carbonate solution and 50 ml brine, dried over MgSO ₄ and filtered. The solvent was removed and the residue was purified by silica gel chromatography using a 4: 1 gradient of PE / EA. Yield: 75%.

Synthesis of 4- (4-Ethoxycarbonyl-thiazol-2-yl) -piperidine-1-carboxylic acid t-butyl ester
4- (4-Ethoxycarbonyl-4,5-dihydro-thiazol-2-yl) -piperidine-1-carboxylic acid tert-butyl ester (1.0 eq, 8.7 mmol) was dissolved in 160 ml of toluene under inert conditions. Dissolved. To this solution was added MnO ₂ (15.0 eq, 130 mmol). The reaction was heated to 70 ° C. with stirring for 5 hours. The mixture was filtered through celite and the filtrate was washed 3 times with 30 ml toluene and ethyl acetate. The organic layer was distilled in vacuum. The residue was purified by silica gel chromatography using a DCM / MeOH 95: 5 gradient. Yield: 30%

・ C-terminal functionalization
4- (4-Ethoxycarbonyl-thiazol-2-yl) -piperidine-1-carboxylic acid tert-butyl ester (1.0 eq, 2.9 mmol) was dissolved in 40 ml dioxane under inert gas. Under stirring, 1.5 ml of 2N NaOH aqueous solution was added dropwise over 10 minutes. Thereafter, the mixture A was stirred at room temperature for 2 hours.

The reaction was neutralized with 2N HCl and the solvent was evaporated in vacuo. The residue was dissolved in 50 ml ethyl acetate. The organic layer was extracted 3 times with 10 ml of 1M citric acid and water, dried over MgSO ₄ and filtered. The solvent was removed and the residue was dried in vacuo. Yield 95%.

  4- (4-Carboxy-thiazol-2-yl) -piperidine-1-carboxylic acid tert-butyl ester (1 eq) dissolved in dry dimethylacetamide (0,03 mmol / ml) under inert conditions did. To this solution was added aryl- or alkylamine (1 eq), diisopropylethylamine (2 eq), and O-benzotriazol-1-yl-N, N, N ', N'-tetramethyluronium hexafluorophosphate (2 eq) was added. The reaction mixture was stirred at room temperature for 12 hours.

The solvent was removed in vacuo and the residue was dissolved in ethyl acetate. The organic layer was extracted 3 times with 1M citric acid, twice with aqueous potassium hydrogen carbonate solution and brine, dried over MgSO ₄ and filtered. The solvent was removed and the residue was purified by silica gel chromatography using a DCM / MeOH 95: 5 gradient. Yield: 40-80%.

・ N-terminal functionalization
The N-protected material was treated with 4 M HCl / dioxane (concentration 0.03 mmol material in 1 mL HCl / dioxane) under inert conditions and stirred at room temperature for 2 hours.

  Removal of the solvent in vacuo yielded the HCl salt of the free amine without further purification.

  The free amino compound (1 eq) was dissolved in dry dimethylacetamide (0,03 mmol / ml) under inert conditions. To this solution was added aryl- or alkyl carboxylic acid (1 eq), diisopropylethylamine (2 eq) and O-benzotriazol-1-yl-N, N, N ', N'-tetramethyluronium hexafluorophosphate (2 eq) were added in this order and the reaction mixture was stirred at room temperature for 12 hours.

The solvent was removed in vacuo and the residue was dissolved in ethyl acetate. The organic layer was extracted three times with 1M citric acid, twice with potassium bicarbonate, and brine, dried over MgSO ₄ and filtered. The solvent was removed and the residue was purified by silica gel chromatography using a DCM / MeOH 95: 5 gradient. Yield: 40-80%.

  Exemplary compounds of formula (Ih) of the present invention include, but are not limited to, the following compounds:

-Compositions containing cytokines and compounds of general formula (Ih)
IFN-β inhibits peripheral blood mononuclear cells (PBMC) proliferation by ConA in peripheral blood mononuclear cells (PBMC) of MS patients and healthy individuals (J Neuroimmunol. 1993; 46 (1-2): 145- 53). We tested the activity of substances of general formula (Ih) in combination with IFN-β in PBMC of healthy donors.

  Human PBMC were stimulated with PHA and growth inhibition was measured in the presence of IFN-β or a substance of formula (Ih). In addition, growth inhibition was quantified in the presence of both substances.

  Furthermore, the activity of the combination of the substance of formula (Ih) and IFN- (on flavivirus-infected cell lines was tested.

Bovine viral diarrhea virus (BVDV) assay:
Preparation of an example of a drug formulation, ie a composition of the invention:
The material was dissolved in DMSO. Peginterferon-alpha (IFN) was provided by the Southern State Research Society and was dissolved in sterilized dH ₂ O. The final DMSO concentration used in the assay is <1% and does not affect the test described.

・ BVDV effectiveness assessment:
Drug-drug interactions were analyzed by testing a wide range of concentrations of the combination of compound and IFN for antiviral and cytotoxic activity. Bovine viral diarrhea virus (BVDV) NADL strain antiviral evaluation test and drug / drug combination analysis were performed as described in Antimicro. Agents Chemother. 2003; 47: 2293-8. BVDV is often used as a hepatitis C virus surrogate model in antiviral drug research (Antiviral Res. 2003; 60: 1-15). Madin-Darby bovine kidney cells are trypsinized, pelleted, counted, and 1 × 10 ^{4 in} cell culture medium in a 96-well flat bottom cell culture plate with a volume of 100 μl per well. Resuspended in / hole. After fixing the cells, the wells were washed and the medium was replaced with complete medium (2% serum) containing various concentrations of compound diluted in the medium. Pre-measured aliquot titers of virus were removed in the freezer (−80 ° C.) immediately prior to each test. The virus was diluted in cell culture medium so that the amount of virus added to each well would give complete cell killing with 6-7 days post infection. The configuration of the test plate for antiviral evaluation is standardized by the Southern State Research Association. Each plate is similar to the test hole (drug + cell + virus), cell control hole (cell only), virus control hole (cell + virus only), drug cytotoxicity hole (cell + drug), and drug colorimetric control Includes holes (drugs only). Drug-drug combination analysis utilizes a grid-like dilution matrix scheme. Samples at 5 substance concentrations were evaluated in combination with samples at 9 IFN concentrations, and antiviral effects and cytotoxicity were monitored by MTS staining at the experimental endpoint.

• MTS staining for cell viability At the end of the test, test plates were stained with the soluble tetrazolium dye MTS (CellTiter Reagent Promega) to determine cell viability and quantify compound toxicity. MTS is metabolically metabolized by mitochondrial enzymes of active cells to produce soluble formazan products, allowing rapid quantitative analysis of cell viability and compound toxicity. MTS is a stable solution and need not be prepared before use. At the end of the test, 10 μl of MTS reagent was added to each well. The microtiter plate was incubated at 37 ° C. for 4 hours. An adhesive sealer is used in place of the lid, the sealed plate is inverted several times to mix with soluble formazan, and the plate is measured spectrophotometrically at 490 nm and 650 nm using a Molecular Devices Vmax plate reader. I read it legally.

・ Data analysis Pharmacological combination test data is analyzed according to the method of Prichard and Shipman (Mac Synergy II. Version 1.0 User's manual. 54pp.) For Mac Synergy II program for data analysis and statistical evaluation. And analyzed. In summary, the Mac Synergy II program calculates the logical additional interaction of a drug based on the Bliss Independence mathematical definition of the expected effect on the drug-drug interaction. The Bliss Independence model assumes that drug action is independent of virus replication based on statistical probabilities.

Logical additive values were calculated from dose response curves for each substrate used individually. This calculated additive surface showing the expected or additive interaction is then subtracted from the experimentally determined dose response surface showing the non-additive active area. The subtracted plane will appear as a 0% inhibition horizontal plane, as calculated above, where the interaction is merely an additive addition. Any peak at the top of this active surface will be synergistic. Similarly, any indentation below this active surface will show antagonism. A 95% confidence interval around the experimental dose response surface is used to statistically evaluate the data, the peak / indentation volume is calculated, and the volume of synergy / antagonism produced is quantified Used for The peak volume observed in the synergy plot (concentration time concentration time percent: typically μM ² %) is calculated by the program. This peak volume is a 3D corresponding area under the 3D dose response surface and is a quantitative measure of synergy or antagonism. This study shows weak synergy or antagonism when synergy or antagonism is between 25-50 μM ² %. A synergistic or antagonizing volume of 50-100 μM ² % shows an intermediate effect, whereas a strong effect is shown when the synergistic or antagonizing volume exceeds 100 μM ² %. Drug synergy or antagonism is followed by replicable clustering of adjacent cells above or below the zero plane of the different plots.

T-lymphocyte proliferation analysis:
Inhibition of stimulated peripheral blood monocytes (PBMC). PBMC were isolated from healthy volunteers blood using the tube ACCUSPIN ^TM System Histopaque ^R -1077, washed, containing 10% fetal calf serum and 2 mM glutamine, in ^{Dulbecco's modified eagles medium, 10 6} cells / ml And resuspended. Cells were activated with 2 μg / ml phytohemaglutinin in the presence of test compound or blank vehicle for 72 hours, and 5′-bromo-2′-desoxyuridine (BrdU) 4 hours before the end of culture. ) Was added for labeling of rapidly proliferating cells. After culture, the cells were separated by centrifugation and the culture supernatant was removed. Incorporated BrdU was quantified by enzyme immunoassay (Roche Diagnostics, Mannheim, Germany).

For the determination of IC ₅₀ values (concentration required for 50% inhibition), at least four inhibitory concentrations were applied. Each data point was recorded three times. The curve was fitted with an appropriate program.

·result:
BVDV assay:
Compound 273, in combination with INF-α, showed high synergistic behavior with respect to binding of their antiviral activity, while showing high antagonistic behavior with respect to their cytotoxic binding. The compounds of formula (Ih) are therefore suitable for the treatment of viral diseases, in particular viral diseases caused by flaviviruses, in combination with cytokines, ie interferons.

PBMC assay:
The compounds of the examples resulted in an inhibition result of more than 50% compared to the control experiment. The average EC50 of the compound was between 3 μM and 40 μM. Co-administration of INF-α produced at least an additive effect on activity. Thus, the compounds of formula (Ih) are ideal for the treatment of inflammatory diseases or diseases associated with T-cells in combination with cytokines, ie interferons.

Claims (14)

(i) cytokines and
(ii) a compound of the general formula (Ih), or a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable prodrug, or a stereoisomer thereof,

Where
A is NR ^{2 ′} , S or O;
t is 0 to 4;
r is 0 or 1;
R ^2a is independently H, OH, SH, NH ₂ , alkyl, cycloalkyl, hydroxyalkyl, haloalkyl, haloalkyloxy, alkoxy, alkylamino, hydroxyalkylamino, halogen, aryl, or heteroaryl;
R ^3a is H, OH, SH, NH ₂ , —C (NR ⁷ ) NR ^{7 ′} R ⁸ , — (CH ₂ ) _p aryl, — (CH ₂ ) _p NR ⁷ R ⁸ , —C (O) NR ⁷ R ⁸ , -N = CR ⁷ R ⁸ , -NR ⁷ C (O) R ⁸ , alkyl, cycloalkyl, hydroxyalkyl, haloalkyl, haloalkyloxy, alkoxy, alkylamino, hydroxyalkylamino, halogen, aryl, or Is heteroaryl;
R ^d is H, halogen, alkyl, -C (NR ⁷ ) NR ^{7 '} R ⁸ ,-(CH ₂ ) _p aryl,-(CH ₂ ) _p NR ⁷ R ⁸ , -C (O) NR ⁷ R ^{8 , -N = CR 7 R 8,} -NR 7 C (O) R 8, cycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, heteroaryl, or aryl;
R ¹ is -C (O) R ^7a , -C (O) CHR ⁷ R ⁸ , -C (O) NR ⁷ R ⁸ , -C (O) OR ⁷ , -R ⁷ C (O) R ⁸ , Or -C (S) R ^7b ;
R ² is H, alkyl, cycloalkyl, heterocycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, or heteroaryl; or R ¹ and R ^{2 taken} together are N-atoms or Together with the C-atom forms a 3-8 membered saturated or at least partially unsaturated monocyclic or polycyclic system, wherein the carbon atoms in at least one or more rings are O, N, And the ring can be substituted by one or more R ⁹ ;
R ³ is H, —C (O) NR ^a R ^b , halogen, alkyl, haloalkyl, aryl, heteroaryl, OH, SH, NR ^{4 ′} OR ^{5 ′} , NH ₂ , hydroxyalkylamino, alkylamino, alkoxy, Is cycloalkyl, hetero-cycloalkyl, hydroxyalkyl, or haloalkyloxy;
R ^a is H, halogen, alkyl, —C (NR ⁷ ) NR ^{7 ′} R ⁸ , — (CH ₂ ) _p aryl, — (CH ₂ ) _p NR ⁷ R ⁸ , aryl, —C (O) NR ⁷ R ⁸ , -N = CR ⁷ R ⁸ , -NR ⁷ C (O) R ⁸ , cycloalkyl, heterocycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, or heteroaryl;
R ^b is independently H, -CN, -OH, -SH, -CO ₂ R ^{4 '} , -C (O) R ^4' , -SO ₂ NR ^{4 '} , -NR ^4' R ^{5 '} , -C (O) NR ⁷ R ⁸ , -SO ₂ -alkyl, -SO ₂ R ^{4 '} , SO ₃ R ^4' , -N = CR ^{4 '} R ^5' , -NR ^{4 '} C (O) R ^4'' , -NR ^{4 '} -CO-haloalkyl, -NO ₂ , -NR ^4' -SO ₂ -haloalkyl, -NR ^{4 '} -SO ₂ -alkyl, -NR ^4' -CO-alkyl, -NR ^{4 '} (CH ₂ ) heteroaryl _p, alkyl, cycloalkyl, alkylamino, alkoxy, alkylthio, halogen, haloalkyl, _{Haroarukirokishi, -O (CH 2) p [} O (CH 2) p] q OCH 3, -C (NR 4 '') NR ^{4 ′} benzimidazolyl, —C (NR ^{4 ″} ) NR ^{4 ′} benzthiazolyl, —C (NR ^{4 ″} ) NR ^{4 ′} benzoxazolyl, hydroxyalkyl, hydroxy-cycloalkyl, hydroxyalkylamino, heterocycloalkyl, Represents aryl or heteroaryl;
R ^{4 ′} , R ^{4 ″} , R ^{5 ′} are independently H, halogen, alkyl, —C (NR ⁷ ) NR ^{7 ′} R ⁸ , — (CH ₂ ) _p aryl, haloalkyl, — (CH ₂ ) _p NR ⁷ R ⁸ , -C (O) NR ⁷ R ⁸ , -N = CR ⁷ R ⁸ , -NR ⁷ C (O) R ⁸ , cycloalkyl, heterocycloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, Is heteroaryl, or aryl;
R ⁷ , R ^{7 ′} , R ⁸ are independently H, halogen, alkyl, cycloalkyl, heterocycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, arylamino heteroaryl, or aryl;
R ^7a is cycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, heteroaryl, or aryl;
R ^7b is H, halogen, alkyl, cycloalkyl, heterocycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, heteroaryl, or aryl;
X is NR ^{2 ′} , O, or S;
Z is N or CR ^{2 ′} ;
R ^{2 ′} is H, alkyl, —C (O) NR ⁷ , —C (O) R ^b , cycloalkyl, heterocycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, heteroaryl, or aryl. Yes;
p is 1 to 6;
q is 1 to 6;
R ⁹ is independently H, -CN, -OH, -SH, alkoxy, alkylthio, -CO ₂ R ^{4 '} , -C (O) R ^4a , -C (O) NR ⁷ R ⁸ , -SO ₂ NR ^{4 '} , -NR ^4' R ^{5 '} , -SO ₂ -alkyl, -SO ₂ R ^4' , SO ₃ R ^{4 '} , -N = CR ^4' R ^{5 '} , -NR ^4' C (O) R ^{4 ''} , -NR ^{4 '} -CO-haloalkyl, -NO ₂ , -NR ^4' -SO ₂ -haloalkyl, -NR ^{4 '} -SO ₂ -alkyl, -NR ^4' -CO-alkyl, -NR ^{4 '} ( CH ₂ ) _p heteroaryl, alkyl, hydroxyalkyl, cycloalkyl, halogen, haloalkyl, alkylamino, —O (CH ₂ ) _p [O (CH ₂ ) _p ] _q OCH ₃ , —C (NR ^{4 ''} ) NR ^{4 'benzimidazolyl,} -C (NR ^4'') NR ^4' benzothiazolyl, -C (NR ^{4 '')} NR ^{4 'benzoxazolyl,} hydroxycycloalkyl, hydroxy - alkylamino, Haroarukirokishi, heterocycloalkyl, - (CH ₂ ) _p NR ⁷ represents COR ⁸ , aryl, or heteroaryl;
R ^4a is H, C ₁ -C ₆ -alkyl, C ₂ -C ₆ -alkenyl, cycloalkyl, haloalkyl, hydroxyalkyl, hydroxyalkylamino, alkylamino, -C (NR ⁷ ) NR ^{7 '} R ⁸ ,- (CH ₂ ) _p aryl,-(CH ₂ ) _p NR ⁷ R ⁸ , -C (O) NR ⁷ R ⁸ , -N = CR ⁷ R ⁸ , -NR ⁷ C (O) R ⁸ , halogen, heteroaryl Or aryl;
here,
If the C ₁ -C ₆ -alkyl group is not otherwise specified, it represents a linear or branched C ₁ -C ₆ -alkyl, which can be optionally substituted with one or more substituents R ′. Yes;
R 'is independently H, -CO ₂ R ", -CONHR", -CR "O, -SO ₂ NR", -NR "-CO-haloalkyl, -NO ₂ , -NR" -SO ₂ -haloalkyl, -NR "-SO ₂ - alkyl, -SO ₂ - alkyl, -NR" -CO- alkyl, -CN, alkyl, cycloalkyl, alkylamino, alkoxy, -OH, -SH, alkylthio, hydroxyalkyl, hydroxyalkylamino Halogen, haloalkyl, haloalkyloxy, aryl, or heteroaryl;
R "is independently H, haloalkyl, hydroxyalkyl, alkyl, cycloalkyl, aryl, or heteroaryl;
C ₂ -C ₆ -alkenyl groups, unless otherwise stated, represent straight-chain or branched C ₂ -C ₆ -alkenyls, which can be optionally substituted by one or more substituents R ′;
An alkyl group, unless stated otherwise, is a linear or branched C ₁ -C ₆ -alkyl group, a linear or branched C ₂ -C ₆ alkenyl group, or a linear or branched C ₂ -C ₆ -alkynyl groups, which can be optionally substituted by one or more substituents R ′, where R ′ is defined above;
Cycloalkyl groups represent non-aromatic ring systems containing from 3 to 8 carbon atoms, in which one or more carbon atoms in the ring can be optionally substituted by one or more substituents R ′, and R ′ Is defined above;
A heterocycloalkyl group represents a non-aromatic ring system comprising 2 to 10 carbon atoms and a heteroatom selected from at least one O, N, and S, wherein one or more carbon atoms in the ring are as defined above. Can be substituted by the group R ′ as defined in
An alkoxy group represents an O-alkyl group, the alkyl group being an alkyl group as defined above;
An alkylthio group represents an S-alkyl group, the alkyl group being an alkyl group as defined above;
A haloalkyl group represents an alkyl group substituted with 1 to 5 halogen atoms, the alkyl group being an alkyl group as defined above;
A hydroxyalkyl group is a HO-alkyl group, which is an alkyl group as defined above;
A haloalkyloxy group represents an alkoxy group substituted with 1 to 5 halogen atoms, the alkyl group being an alkyl group as defined above;
A hydroxyalkylamino group represents a (HO-alkyl) ₂ -N- group or a HO-alkyl-NH- group, the alkyl group being an alkyl group as defined above;
An alkylamino group represents an HN-alkyl group or an N-dialkyl group; the alkyl group is an alkyl group as defined above;
The halogen group is fluorine, chlorine, bromine or iodine;
Aryl groups are aromatic groups having 5 to 15 carbon atoms, which atoms can be substituted with one or more R ′, where R ′ is as defined above;
An arylamino group represents an HN-aryl group or an N-diaryl group, said aryl group being an aryl group as defined above;
A heteroaryl group is a 5- to 10-membered heterocyclic ring system containing at least one heteroatom selected from O, N, and S, wherein the heterocyclic group is fused with another ring. Each of the heterocyclic group or the fused ring may be substituted by one or more R ′ as defined above;
A pharmaceutical composition comprising In the formula (Ih), each of the following substituents or a combination of the following substituents:
X is S or O, and
The composition of claim 1, wherein R ^3a is H. The composition of claim 1 or 2, wherein X is S. The composition of claims 1-3, wherein Z is CR ^{2 '} and R ¹ and R ² are taken together to form a saturated 6-membered ring with the C-atom. The composition of claims 1 to 3 wherein Z is N and R ¹ and R ² together form a saturated 6-membered ring with the N-atom. R ⁹ is heteroaryl, selected from the group consisting of aryl and benzyl, The composition of claim 1 5. R ⁹ is, thienopyrimidine, quinazoline, purine, chosen from the group of pyrazolopyrimidine or thoria sol pyrimidine The composition of claim 6. t is preferably 0, and
A composition according to any preceding claim, wherein r is preferably 1. A composition according to any of the preceding claims, wherein R ^2a is selected from the group of OH, alkyl, aryl and heteroaryl. The composition of any of the preceding claims, wherein said cytokine (i) is selected from the group of (a) autocrine cytokines, (b) paracrine cytokines, and (c) endocrine cytokines. The cytokine is the following cytokine family:
(a) IL-2 subfamily,
(b) the interferon (IFN) subfamily, and
(c) consisting of the IL-10 subfamily
(i) four α-helix cytokine families;
(ii) IL-1 family,
(iii) IL-17 family, and
(iv) chemokines,
11. The composition of claim 10, wherein the composition is selected from: The cytokine is
(i) IL-2 family including IL-4, IL-7, IL-9, IL-15 and IL-21
(ii) interferon family including type I interferon, type II interferon, and type III interferon, and
(iii) tumor necrosis factor (TNF), including TNF-α (alpha) and TNF-β (beta),
12. The composition of claims 10 and 11 selected from the group of 13. The composition of claim 12, wherein the cytokine is selected from the group of IFN-α (alpha), IFN-β (beta), IFN-γ (gamma), IL-2 and TNF. Use of the composition of claims 1 to 13 for the manufacture of a medicament.

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