COMPOUNDS WITH A BENZO[A]CARBAZOLE STRUCTURE AND USE THEREOF

Abstract

Disclosed are compounds of general formula (I), and their use for the treatment and diagnosis of degenerative disorders characterized by high cell proliferation and/or tissue degeneration.

Claims

1. Compounds of general formula (I): ##STR00176## wherein n is an integer ranging from 0-12; m is an integer ranging from 0-2; the phenyl rings A e D condensed in the tetracyclic system A-D are optionally substituted, both or only the ring A o only the ring D, in any of the substitutable, by a chain of formula II o II: ##STR00177## wherein the chain of formula II may be represented by an amino acid, natural o non-natural, of straight type of formula III (with T absent) or cyclic type of formula IV (with T present); or by a peptide sequence, containing amino acids natural and/or non-natural of type (III) and (IV), of formula (V); o by a peptide sequence of formula (VI) ending with an amino acid of type (III) or (IV); or by a sequence of formula (VII) or (VIII): ##STR00178## R is selected from the group consisting of hydrogen or a group R.sub.1 or a group G wherein G is a carbon atom also halogenated or poly-halogenated with atoms of F or Cl or Br or I, or a carbon atom of a monocyclic o bicyclic aryl, or a carbon or a nitrogen atom being part of an aromatic or non-aromatic heterocyclic system selected from the group consisting of pyrrole, pyrrolidine, 3-pyrroline, 2H-pyrrole, 2-pyrroline, indole, isoindole, 3H-indole, indolizine, indoline, furan, benzofuran, isobenzofuran, 2H-pyran, 4H-pyran, benzo[b]thiophene, thiophene, pyridine, piperidine, 4H-quinolizine, isoquinoline, quinoline, tetrahydroquinoline, 1,8-naphthyridine, acridine, oxazole, isoxazole, benzoxazole, benzothiazole, isothiazole, triazole, imidazole, 2-imidazole, imidazolidine, tetrazole, 1,2,3-triazole, 1,2,4-triazole, 1,2,3-oxadiazole, benzimidazole, purine, 1,4-dioxane, 1,3-dioxolane, 1,3-dithiane, 1,4-dithiane, 1,3,5-trithiane, morpholine, thiomorpholine, phenothiazine, pyrazole, 2-pyrazoline, pyrazolidine, quinazoline, cinnoline, pyrimidine, pyrazine, pteridine, phthalazine, 1,2,4-triazine, 1,3,5-triazine, pyridazine, piperazine, quinoxaline, phenazine, 1H-indazole; or R is a C.sub.1-C.sub.10 carbon atoms chain, saturated or unsaturated, straight or branched, optionally substituted with a substituent selected from R.sub.1, G, a hydroxyl, O-alkyl, ONO.sub.2, O-G, phenyl, substituted phenyl, heteroaryl, substituted heteroaryl, NH-G, NG.sub.2, NR.sub.7R.sub.8, NO-G, NHO-G, COOH, (CH.sub.2).sub.pCOOH, (CHR.sub.2).sub.pCOOH, (CH.sub.2).sub.pCONHR, (CHR.sub.2).sub.pCONHR, wherein p can be a number from 0 to 12 and wherein R.sub.7 and R.sub.8 are independently selected from hydrogen, all the meanings of R.sub.1, all the meanings of R.sub.2; or R is a C.sub.1-C.sub.6 carbon atoms chain, saturated or unsaturated, straight or branched, optionally substituted with aryl, CO alkyl, CO-aryl, CO-heteroaryl; CONH-alkyl, CONH-alkyl-ONO.sub.2; CONH-acyl; CONH-acyl-ONO.sub.2, CONH-aryl, SO.sub.2-alkyl, SO.sub.2NH.sub.2, SO.sub.2NH-alkyl, SO.sub.2NH-aryl, wherein aryl may be phenyl, substituted, phenyl, heteroaryl or substituted heteroaryl, and wherein R is selected from alkyl, alkyl-ONO.sub.2, aryl, acyl e acylaryl, wherein aryl may be phenyl, substituted, phenyl, heteroaryl or substituted heteroaryl, or as indicated in the meanings of G, when m=2 the groups R are 2 and they are 2 substituents, the same or different, selected from the meanings of R; in some cases two groups R can form together a ring which has as closure the bearing carbon of R with the proviso that when m is 2, T is absent; R.sub.1 is selected from H, F, Cl, Br, I, -aryl, -heteroaryl, R, NO.sub.2, NH.sub.2, NHCH.sub.3, NH-alkyl, NH-aryl, NH-heteroaryl, NHR, N(R).sub.2, NRR.sub.1, N(CH.sub.2).sub.2, N(CH.sub.2).sub.3, N(CH.sub.2).sub.4, N(CH.sub.2).sub.5, N(CH.sub.2).sub.4O, N(CH.sub.2).sub.4S(O).sub.m, N(CH.sub.2).sub.4NR, N(CH.sub.2).sub.4NR.sub.1, NHCOCH.sub.3, NHCO-alkyl, NHCO-cycloalkyl, NHCO-aryl, NHCO heteroaryl, NHCOR, NHSO.sub.2CH.sub.3, NHSO.sub.2-alkyl, NHSO.sub.2-cycloalkyl, NHSO.sub.2-aryl, NHSO.sub.2-heteroaryl, NHSO.sub.2R, SCH.sub.3, SR, SO.sub.2CH.sub.3, SO.sub.2-alkyl, SO.sub.2-cycloalkyl, SO.sub.2-aryl, SO.sub.2-heteroaryl, SO.sub.2R, SO.sub.2NH.sub.2, SO.sub.2NHCH.sub.3, SO.sub.2NH-alkyl, SO.sub.2NH-cycloalkyl, SO.sub.2NH-aryl, SO.sub.2NH-heteroaryl, SO.sub.2NHR, SO.sub.2NHCOCH3, SO.sub.2NHCO-alkyl, SO.sub.2NHCO-aryl, SO.sub.2NHCO-heteroaryl, SO.sub.2NHCO-cycloalkyl, C(CH).sub.4N (e.g. 2-pyridine, 3-pyridine, 4-pyridine), C(CH).sub.3O (e.g. 2-furan, 3-furan), C(CH).sub.3S (e.g. 2-thiophene, 3-thiophene), CH(CH.sub.2)O (e.g. oxirane), CH(CH.sub.2)S (e.g. thiiran), CO.sub.2H, CO.sub.2CH.sub.3, CO.sub.2-alkyl, CO.sub.2-aryl, CO.sub.2-heteroaryl, CO.sub.2-cycloalkyl, CO.sub.2R, CONHCH.sub.3, CONH-alkyl, CONH-aryl, CONH-heteroaryl, CONH-cycloalkyl, CONHR, CONRCH.sub.3, CONRR, CONHSO.sub.2CH.sub.3, CONHSO.sub.2-alkyl, OCH.sub.2-alkyl, O(CH.sub.2)n-R, O-acyl, O-aryl, O-heteroaryl, O-cycloalkyl, OR, OCH.sub.2OCH.sub.3, OCH.sub.2OCH.sub.2CH.sub.3, OCH.sub.2(OCH.sub.2CH.sub.2).sub.nOCH3, O(CH.sub.2).sub.nNH-alkyl, O(CH.sub.2).sub.nN-(alkyl)2, O(CH.sub.2).sub.nNH-cycloalkyl with ring from 4 to 6 carbon atoms, O(CH.sub.2).sub.nR with the R group as defined above, OCH.sub.2 (NH CH.sub.2CH.sub.2)n-OCH.sub.3, OCH.sub.2(CH.sub.2).sub.nCO(NHCH.sub.2CH.sub.2).sub.nOCH.sub.3, OCH.sub.2(CH.sub.2).sub.nSO.sub.2(NHCH.sub.2CH.sub.2).sub.nOCH.sub.3, N(CH.sub.2CH.sub.2).sub.2N(CH.sub.2CH.sub.2).sub.nNH, CH.sub.3N(CH.sub.2CH.sub.2).sub.2N(CH.sub.2CH.sub.2).sub.n, CH.sub.3 (CH.sub.2).sub.nCON(CH.sub.2CH.sub.2).sub.2N(CH.sub.2CH.sub.2).sub.n, CH.sub.3 (CH.sub.2).sub.nSO.sub.2N(CH.sub.2CH.sub.2).sub.2N(CH.sub.2CH.sub.2).sub.n, O(CH.sub.2CH.sub.2).sub.2N(CH.sub.2CH.sub.2).sub.n, monocyclic or bicyclic aryl, or an aromatic or non-aromatic heterocyclic system selected from the meanings of R; M is selected from O, CH.sub.2, CHR.sub.2, CCHR.sub.2, C(R.sub.2).sub.m, CHOH, CHOR.sub.2, CCHOR.sub.2, CO, CONR.sub.2, NH, NR.sub.2, NOH, NOR.sub.2, NR.sub.2CO, S, SO, S(O).sub.2, wherein R.sub.2 is independently selected from the meanings of R, or R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8; M.sub.1 is independently selected from the meanings of M, as defined above, or absent; R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8, the same or different, are independently selected from the meanings of R, or R, R.sub.1; P is selected from O, Q, CH.sub.2, CHR.sub.2, C (R.sub.2).sub.m, CHOH, CHOR.sub.2, CHO-acyl, NH, N-acyl, NR.sub.2, NOH, NOR.sub.2, NO-acyl, wherein R.sub.2 is independently selected from the meanings of R, or R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8; P.sub.1 is independently selected from the meanings of P, as defined above, or absent; Q is selected from H, R.sub.2, OR.sub.2, O-acyl, NH.sub.2, NHR.sub.2, N (R.sub.2).sub.m, NH-acyl, NHCOOCH.sub.2Bn, NHCOO-t-Bu, NHCOOR, NHOH, NHOR.sub.2, N(R.sub.2)OR.sub.2, NHO-acyl, wherein R.sub.2 is independently selected from the meanings of R, or R, R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8; or Q is a beta-lactam structure of formula (IX), (IX), (IX): ##STR00179## wherein Q is selected from H, R.sub.2, OR.sub.2, O-acyl, NH.sub.2, NHR.sub.2, N(R.sub.2).sub.m, NH-acyl, NHCOOCH.sub.2Bn, NHCOO-t-Bu, NHCOOR, NHOH, NHOR.sub.2, N(R.sub.2)OR.sub.2, NHO-acyl, wherein R.sub.2 is independently selected from the meanings of R, or R, R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8; Q.sub.1 is independently selected from the meanings of Q, as defined above, or absent; T is a saturated or containing unsaturation cyclic ring, consisting of 3 to 8 atoms linked together and containing carbon atoms and/or also N, O or S(O).sub.m or also substituted, as in the defined meanings, by R, R, R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8 or by X, M, P, W or Z or as defined in the meanings of the cyclic systems of G for R and/or R.sub.1; T.sub.1 is independently selected from the meanings as T, as defined above, or absent; X is selected from groups CH.sub.2, CH(R), C(R), O, S(O).sub.m, N, N(R), C(O), C(P), N(R) CO, CON(R), wherein R is H is one of the meanings defined for R, R and for R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8; W is selected from H, or from the meanings of T or Z or R, or also of R, R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8; Z is selected from H, CH.sub.3, CH.sub.2OCOCH.sub.3, or, taken together with the carbon atom to which it is linked, Z is selected from CH.sub.2, CH(R.sub.2), C(O), C(P); or Z is selected from the meanings of R, or of R, R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8, or also from the meanings of M, P.

2. Compounds according to claim 1, wherein the compounds of formula (I) are characterized by a substructure of formula (X): ##STR00180## wherein m, X, R.sub.1, R.sub.2, R.sub.3 are as defined in claim 1.

3. Compounds according to claim 2, wherein the compounds of formula (X) are characterized by a substructure of formula (XI) or (XII): ##STR00181## wherein ##STR00182## is a substituent of formula IX, IX or IX.

4. Compounds according to claim 3, wherein the compounds of formula (XII) are characterized by a substructure of formula (XIII), (XIV) or (XV): ##STR00183##

5. Compounds according to claim 2, wherein the compounds of formula (X) have a formula selected from the following: ##STR00184## ##STR00185## ##STR00186## ##STR00187##

6. Compounds according to claim 3, wherein the compounds of formula (XI) have a formula selected from the following: ##STR00188## ##STR00189## ##STR00190## ##STR00191##

7. Compounds according to claim 4, wherein the compounds of formula (XIII) have a formula selected from the following: ##STR00192## ##STR00193## ##STR00194## ##STR00195##

8. Compounds according to claim 4, wherein the compounds of formula (XIV) have a formula selected from the following: ##STR00196## ##STR00197## ##STR00198## ##STR00199##

9. Compounds according to claim 4, wherein the compounds of formula (XV) have a formula selected from the following: ##STR00200## ##STR00201## ##STR00202## ##STR00203##

10. Compounds according to claim 1, wherein the compounds are selected from the group consisting of: TABLE-US-00003 # # substructure Abbreviation Structure XIc XIl RA1 XIa RA2 XIr RA3 Xa GA17M8 Xa GA17M10 XIb GA11S XIb RM66 XIb RM58 XIb RM70 XIb RM85 XI'd RA6 XI'd RA7 XI'd RA9 XI'd RA11 XI'd RA12 XI'd RA13 XI'g RA14 X'c RA15 X'c RA16 XI'c RA17 XI'c RA18 XI'c RA19 XI'h RA20 X'e RA21 XI'e RA22 XIV'a GA09a XIV'a' GA09a' XIV'a GA09b XIV'a' GA09b' XIV'a RM36 XIV'a RM37 XIV'a' RM53 XIV'a RM47 XIV'd RA23 XIV'd RA24 XIV'd RA25 XV'e RA26 XV'd RA27 XV'd RA28 XV'g' RA29 XIIId RA30 XIV'c RA31 XIV'c RA32 XIV'c RA33 XIII'c RA34 XV'c RA35 XIV'e RA36 XIV'g' RA37 XIV'g' RA38 XIV'g RA39 XIVr RA40 XIVc RA41 XVr RA42 XIVc RA43 XIIIr RA44

11. A method for treatment or diagnosis of degenerative pathologies characterized by high cell proliferation and/or tissue degeneration, comprising applying an effective amount of the compound of claim 1.

12. Compounds according to claim 2, wherein the compounds are selected from the group consisting of: TABLE-US-00004 # # substructure Abbreviation Structure XIc XII RA1 XIa RA2 XIr RA3 Xa GA17M8 Xa GA17M10 XIb GA11S XIb RM66 XIb RM58 XIb RM70 XIb RM85 XI'd RA6 XI'd RA7 XI'd RA9 XI'd RA11 XI'd RA12 XI'd RA13 XI'g RA14 X'c RA15 X'c RA16 XI'c RA17 XI'c RA18 XI'c RA19 XI'h RA20 X'e RA21 XI'e RA22 XIV'a GA09a XIV'a' GA09a' XIV'a GA09b XIV'a' GA09b' XIV'a RM36 XIV'a RM37 XIV'a' RM53 XIV'a RM47 XIV'd RA23 XIV'd RA24 XIV'd RA25 XV'e RA26 XV'd RA27 XV'd RA28 XV'g' RA29 XIIId RA30 XIV'c RA31 XIV'c RA32 XIV'c RA33 XIII'c RA34 XV'c RA35 XIV'e RA36 XIV'g' RA37 XIV'g' RA38 XIV'g RA39 XIVr RA40 XIVc RA41 XVr RA42 XIVc RA43 XIIIr RA44

13. Compounds according to claim 3, wherein the compounds are selected from the group consisting of: TABLE-US-00005 # # substructure Abbreviation Structure XIc XII RA1 XIa RA2 XIr RA3 Xa GA17M8 Xa GA17M10 XIb GA11S XIb RM66 XIb RM58 XIb RM70 XIb RM85 XI'd RA6 XI'd RA7 XI'd RA9 XI'd RA11 XI'd RA12 XI'd RA13 XI'g RA14 X'c RA15 X'c RA16 XI'c RA17 XI'c RA18 XI'c RA19 XI'h RA20 X'e RA21 XI'e RA22 XIV'a GA09a XIV'a' GA09a' XIV'a GA09b XIV'a' GA09b' XIV'a RM36 XIV'a RM37 XIV'a' RM53 XIV'a RM47 XIV'd RA23 XIV'd RA24 XIV'd RA25 XV'e RA26 XV'd RA27 XV'd RA28 XV'g' RA29 XIIId RA30 XIV'c RA31 XIV'c RA32 XIV'c RA33 XIII'c RA34 XV'c RA35 XIV'e RA36 XIV'g' RA37 XIV'g' RA38 XIV'g RA39 XIVr RA40 XIVc RA41 XVr RA42 XIVc RA43 XIIIr RA44

14. Compounds according to claim 4, wherein the compounds are selected from the group consisting of: TABLE-US-00006 # # substructure Abbreviation Structure XIc XII RA1 XIa RA2 XIr RA3 Xa GA17M8 Xa GA17M10 XIb GA11S XIb RM66 XIb RM58 XIb RM70 XIb RM85 XId RA6 XId RA7 XId RA9 XId RA11 XId RA12 XId RA13 XIg RA14 Xc RA15 Xc RA16 XIc RA17 XIc RA18 XIc RA19 XIh RA20 Xe RA21 XIe RA22 XIVa GA09a XIVa GA09a XIVa GA09b XIVa GA09b XIVa RM36 XIVa RM37 XIVa RM53 XIVa RM47 XIVd RA23 XIVd RA24 XIVd RA25 XVe RA26 XVd RA27 XVd RA28 XVg RA29 XIIId RA30 XIVc RA31 XIVc RA32 XIVc RA33 XIIIc RA34 XVc RA35 XIVe RA36 XIVg RA37 XIVg RA38 XIVg RA39 XIVr RA40 XIVc RA41 XVr RA42 XIVc RA43 XIIIr RA44

15. Compounds according to claim 5, wherein the compounds are selected from the group consisting of: TABLE-US-00007 # # substructure Abbreviation Structure XIc XII RA1 XIa RA2 XIr RA3 Xa GA17M8 Xa GA17M10 XIb GA11S XIb RM66 XIb RM58 XIb RM70 XIb RM85 XId RA6 XId RA7 XId RA9 XId RA11 XId RA12 XId RA13 XIg RA14 Xc RA15 Xc RA16 XIc RA17 XIc RA18 XIc RA19 XIh RA20 Xe RA21 XIe RA22 XIVa GA09a XIVa GA09a XIVa GA09b XIVa GA09b XIVa RM36 XIVa RM37 XIVa RM53 XIVa RM47 XIVd RA23 XIVd RA24 XIVd RA25 XVe RA26 XVd RA27 XVd RA28 XVg RA29 XIIId RA30 XIVc RA31 XIVc RA32 XIVc RA33 XIIIc RA34 XVc RA35 XIVe RA36 XIVg RA37 XIVg RA38 XIVg RA39 XIVr RA40 XIVc RA41 XVr RA42 XIVc RA43 XIIIr RA44

16. Compounds according to claim 6, wherein the compounds are selected from the group consisting of: TABLE-US-00008 # # substructure Abbreviation Structure XIc XII RA1 XIa RA2 XIr RA3 Xa GA17M8 Xa GA17M10 XIb GA11S XIb RM66 XIb RM58 XIb RM70 XIb RM85 XId RA6 XId RA7 XId RA9 XId RA11 XId RA12 XId RA13 XIg RA14 Xc RA15 Xc RA16 XIc RA17 XIc RA18 XIc RA19 XIh RA20 Xe RA21 XIe RA22 XIVa GA09a XIVa GA09a XIVa GA09b XIVa GA09b XIVa RM36 XIVa RM37 XIVa RM53 XIVa RM47 XIVd RA23 XIVd RA24 XIVd RA25 XVe RA26 XVd RA27 XVd RA28 XVg RA29 XIIId RA30 XIVc RA31 XIVc RA32 XIVc RA33 XIIIc RA34 XVc RA35 XIVe RA36 XIVg RA37 XIVg RA38 XIVg RA39 XIVr RA40 XIVc RA41 XVr RA42 XIVc RA43 XIIIr RA44

17. Compounds according to claim 7, wherein the compounds are selected from the group consisting of: TABLE-US-00009 # # substructure Abbreviation Structure XIc XII RA1 XIa RA2 XIr RA3 Xa GA17M8 Xa GA17M10 Xlb GA11S Xlb RM66 Xlb RM58 Xlb RM70 Xlb RM85 XId RA6 XId RA7 XId RA9 XId RA11 XId RA12 XId RA13 XIg RA14 Xc RA15 Xc RA16 XIc RA17 XIC RA18 XIC RA19 XIh RA20 Xe RA21 Xe RA22 XIVa GA09a XIVa GA09a XIVa GA09b XIVa GA09b XIVa RM36 XIVa RM37 XIVa RM53 XIVa RM47 XIVd RA23 XIVd RA24 XIVd RA25 XVe RA26 XVd RA27 XVd RA28 XVg RA29 XIIId RA30 XIVc RA31 XIVc RA32 XIVc RA33 XIIIc RA34 XVc RA35 XIVe RA36 XIVg RA37 XIVg RA38 XIVg RA39 XIVr RA40 XIVc RA41 XVr RA42 XIVc RA43 XIIIr RA44

18. Compounds according to claim 8, wherein the compounds are selected from the group consisting of: TABLE-US-00010 # # substructure Abbreviation Structure XIc XII RA1 XIa RA2 XIr RA3 Xa GA17M8 Xa GA17M10 XIb GA11S XIb RM66 XIb RM58 XIb RM70 XIb RM85 XId RA6 XId RA7 XId RA9 XId RA11 XId RA12 XId RA13 XIg RA14 Xc RA15 Xc RA16 XIc RA17 XIc RA18 XIc RA19 XIh RA20 Xe RA21 XIe RA22 XIVa GA09a XIVa GA09a XIVa GA09b XIVa GA09b XIVa RM36 XIVa RM37 XIVa RM53 XIVa RM47 XIVd RA23 XIVd RA24 XIVd RA25 XVe RA26 XVd RA27 XVd RA28 XVg RA29 XIIId RA30 XIVc RA31 XIVc RA32 XIVc RA33 XIIIc RA34 XVc RA35 XIVe RA36 XIVg RA37 XIVg RA38 XIVg RA39 XIVr RA40 XIVc RA41 XVr RA42 XIVc RA43 XIIIr RA44

19. Compounds according to claim 9, wherein the compounds are selected from the group consisting of: TABLE-US-00011 # # substructure Abbreviation Structure XIC XII RA1 XIa RA2 XIr RA3 Xa GA17M8 Xa GA17M10 XIb GA11S XIb RM66 XIb RM58 XIb RM70 XIb RM85 XId RA6 XId RA7 XId RA9 XId RA11 XId RA12 XId RA13 XIg RA14 Xc RA15 Xc RA16 XIc RA17 XIc RA18 XIc RA19 XIh RA20 Xe RA21 XIe RA22 XIVa GA09a XIVa GA09a XIVa GA09b XIVa GA09b XIVa RM36 XIVa RM37 XIVa RM53 XIVa RM47 XIVd RA23 XIVd RA24 XIVd RA25 XVe RA26 XVd RA27 XVd RA28 XVg RA29 XIIId RA30 XIVc RA31 XIVc RA32 XIVc RA33 XIIIc RA34 XVc RA35 XIVe RA36 XIVg RA37 XIVg RA38 XIVg RA39 XIVr RA40 XIVc RA41 XVr RA42 XIVc RA43 XIIIr RA44

20. A method for treatment or diagnosis of degenerative pathologies characterized by high cell proliferation and/or tissue degeneration, comprising applying an effective amount of the compound of claim 2.

Description

LIST OF FIGURES

[0006] FIG. 1 shows the stabilisation of p53 in the presence of RM37 or Nut-3.

[0007] FIG. 2 shows the effect of RM37 and Nut-3 on activation of the gene transactivation function of p53.

[0008] FIG. 3 shows the effects of RM37 on stabilisation of the intracellular levels of protein p5.

[0009] FIG. 4 shows the in vitro antitumoral effect of RM37 compared with Nut-3 (dead cell count).

[0010] FIG. 5 shows the in vitro antitumoral effect of RM37 compared with Nut-3 (live cell count).

[0011] FIG. 6 shows the effect of RM37 on the cell cycle.

[0012] FIG. 7 and FIG. 8 show the effect of RM37 on cell apoptosis.

SUMMARY OF THE INVENTION

[0013] The present invention relates to compounds of general formula (I):

##STR00004##

[0014] as defined in the detailed description below.

[0015] The present invention also relates to the use of the compounds with general formula (I) for the treatment and diagnosis of degenerative disorders characterised by high cell proliferation and/or tissue degeneration.

DETAILED DESCRIPTION OF THE INVENTION

[0016] It has now surprisingly been found that compounds of general formula (I) as hereinafter described, which are functionalisable on their central carbazole core, can be used not only as therapeutic agents but also as diagnostic probes in endogenous or exogenous degenerative disorders (ie. those induced by infectious agents such as bacteria, viruses, protozoa or fungi) characterised by high cell proliferation and/or tissue degeneration.

[0017] The compounds of the invention have no endogenous or exogenous protease-inhibiting activities, such as the ability to inhibit activity on bacterial peptidases (e.g. bacterial transpeptidases and beta-lactamases), and have no modulating activity on steroid receptors. Their action takes place solely on the cell cycle, by modulating the functions of oncogenic proteins. Another aspect is modulation of the tumour-suppressor functions of genes, such as that of oncogenic protein p53 or its mutated forms, and of their modulators. The compounds of the invention modulate, for therapeutic purposes, the processes of arrest of the cell cycle and apoptosis in degenerative cells characterised by alteration of the gene array or by oncogenic effects due to sequestration of control proteins; in the latter case, for example, characterised by the MDM2-p53 or MDMX-p53 interaction. For example, the compounds can bind MDM2 or MDMX and modulate the activity of p53; this can lead to a control of the cell cycle with activating/inhibiting effects on proliferation and/or programmed cell life/death.

[0018] As regards the diagnostic aspect, the compounds of the invention comprise one or more labelled residues with one or more residues for imaging, by means of which they can specifically target cells characterised by high proliferation in degenerative tissues or infectious tissues generated by pathogens (bacteria, viruses, protozoa or fungi) and characteristic of all pathological forms associated with oncogenic imbalance. The disorders which can be treated or diagnosed with the compounds of the invention are cancers of various kinds, and other disorders such as infectious diseases caused by pathogens wherein physiological homeostatic tissue control has been lost, and control of proliferative activity and cell apoptosis is important.

[0019] The subject of the present invention is compounds of general formula (I):

##STR00005##

[0020] wherein

[0021] n is an integer between 0 and 12;

[0022] m is an integer between 0 and 2;

[0023] the phenyl rings A and D condensed in tetracyclic system A-D can be optionally substituted (ring A only, ring D only or both), in any of the substitutable positions, with the chain of formula II or II:

##STR00006##

[0024] The chain of formula II can consist of a natural or non-natural amino acid, which may be the linear type of formula III (with T absent) or the cyclic type of formula IV (with T present); or of a peptide sequence, containing natural and/or non-natural amino acids type (III) and (IV), of formula (V); or of a peptide sequence of formula (VI) terminating with an amino acid of type (III) or (IV); or of a sequence of formula (VII) or (VIII):

##STR00007##

[0025] R is selected from the group consisting of hydrogen or an R.sub.1 group or a G group wherein G can be a carbon atom, which may be halogenated or polyhalogenated with F or Cl or Br or I atoms, or also a carbon atom of a monocyclic or bicyclic aryl, or a carbon or nitrogen atom belonging to an aromatic or non-aromatic heterocyclic system selected from the group comprising pyrrole, pyrrolidine, 3-pyrroline, 2H-pyrrole, 2-pyrroline, indole, isoindole, 3H-indole, indolizine, indoline, furan, benzofuran, isobenzofuran, 2H-pyran, 4H-pyran, benzo[b]thiophene, thiophene, pyridine, piperidine, 4H-quinolizine, isoquinoline, quinolines, tetrahydroquinoline, 1,8-naphthyridine, acridine, oxazole, isoxazole, benzoxazole, benzothiazole, isothiazole, thiazole, imidazole, 2-imidazole, imidazolidine, tetrazole, 1,2,3-triazole, 1,2,4-triazole, 1,2,3-oxadiazole, benzoimidazole, purine, 1,4-dioxane, 1,3-dioxolane, 1,3-dithiane, 1,4-dithiane, 1,3,5-trithiane, morpholine, thiomorpholine, phenothiazine, pyrazole, 2-pyrazoline, pyrazolidine, quinazoline, cinnoline, pyrimidine, pyrazine, pteridine, phthalazine, 1,2,4-triazine, 1,3,5-triazine, pyridazine, piperazine, quinoxaline, phenazine and 1H-indazole;

[0026] R can also be a chain of saturated or unsaturated, straight or branched C.sub.1-C.sub.10 carbon atoms optionally substituted with a substituent selected from R.sub.1, G, a hydroxyl, O-alkyl, ONO.sub.2, O-G, phenyl, substituted phenyl, heteroaryl, substituted heteroaryl, NH-G, NG.sub.2, NR.sub.7R.sub.8, NO-G, NHO-G, COOH, (CH.sub.2).sub.pCOOH, (CHR.sub.2).sub.pCOOH, (CH.sub.2).sub.pCONHR, (CHR.sub.2).sub.pCONHR, wherein p can be a number between 0 and 12 and wherein R.sub.7 and R.sub.8 are selected independently from hydrogen, all the meanings of R.sub.1, and all the meanings of R.sub.2;

[0027] or R is a chain of saturated or unsaturated, straight or branched C.sub.1-C.sub.6 carbon atoms optionally substituted with aryl, CO-alkyl, CO-aryl, CO-heteroaryl; CONH alkyl, CONH-alkyl-ONO.sub.2; CONH-acyl; CONH-acyl-ONO.sub.2, CONH-aryl, SO.sub.2-alkyl, SO.sub.2NH.sub.2, SO.sub.2NH-alkyl, SO.sub.2NH-aryl, wherein aryl can be phenyl, substituted phenyl, heteroaryl or substituted heteroaryl, and wherein R is selected from alkyl, alkyl-ONO.sub.2, aryl, acyl and acylaryl, wherein aryl can be phenyl, substituted phenyl, heteroaryl or substituted heteroaryl or as indicated in the meanings of G, when m=2 there are 2R groups and there can be two substituents which are the same or different, selected from the meanings of R; in some of said substitutions the two R groups can jointly constitute a cycle whose closing is the carbon bearing R, on the proviso that when m is 2, T is absent;

[0028] R.sub.1 is selected from H, F, Cl, Br, I, -aryl, -heteroaryl, R, NO.sub.2, NH.sub.2, NHCH.sub.3, NH-alkyl, NH-aryl, NH-heteroaryl, NHR, N(R).sub.2, NRR.sub.1, N(CH.sub.2).sub.2, N(CH.sub.2).sub.3, N(CH.sub.2).sub.4, N(CH.sub.2).sub.5, N(CH.sub.2).sub.4O, N(CH.sub.2).sub.4S(O).sub.m, N(CH.sub.2).sub.4NR, N(CH.sub.2).sub.4NR.sub.1, NHCOCH.sub.3, NHCO-alkyl, NHCO-cycloalkyl, NHCO-aryl, NHCO-heteroaryl, NHCOR, NHSO.sub.2CH.sub.3, NHSO.sub.2-alkyl, NHSO.sub.2-cycloalkyl, NHSO.sub.2-aryl, NHSO.sub.2-heteroaryl, NHSO.sub.2R, SCH.sub.3, SR, SO.sub.2CH.sub.3, SO.sub.2-alkyl, SO.sub.2-cycloalkyl, SO.sub.2-aryl, SO.sub.2-heteroaryl, SO.sub.2R, SO.sub.2NH.sub.2, SO.sub.2NHCH.sub.3, SO.sub.2NH-alkyl, SO.sub.2NH-cycloalkyl, SO.sub.2NH-aryl, SO.sub.2NH-heteroaryl, SO.sub.2NHR, SO.sub.2NHCOCH.sub.3, SO.sub.2NHCO-alkyl, SO.sub.2NHCO-aryl, SO.sub.2NHCO-heteroaryl, SO.sub.2NHCO-cycloalkyl, C(CH).sub.4N (e.g. 2-pyridine, 3-pyridine, 4-pyridine), C(CH).sub.3O (e.g. 2-furan, 3-furan), C(CH).sub.3S (e.g. 2-thiophene, 3-thiophene), CH(CH.sub.2)O (e.g. oxirane), CH(CH.sub.2)S (e.g. thiirane), CO.sub.2H, CO.sub.2CH.sub.3, CO.sub.2-alkyl, CO.sub.2-aryl, CO.sub.2-heteroaryl, CO.sub.2-cycloalkyl, CO.sub.2R, CONHCH.sub.3, CONH-alkyl, CONH-aryl, CONH-heteroaryl, CONH-cycloalkyl, CONHR, CONRCH.sub.3, CONRR, CONHSO.sub.2CH.sub.3, CONHSO.sub.2-alkyl, OCH.sub.2-alkyl, O(CH.sub.2).sub.nR, O-acyl, O-aryl, O-heteroaryl, O-cycloalkyl, OR, OCH.sub.2OCH.sub.3, OCH.sub.2OCH.sub.2CH.sub.3, OCH.sub.2(OCH.sub.2CH.sub.2).sub.nOCH.sub.3, O(CH.sub.2).sub.nNH-alkyl, O(CH.sub.2).sub.nN-(alkyl).sub.2, O(CH.sub.2).sub.nNH cycloalkyl with cycle from 4 to 6 carbon atoms, O(CH.sub.2).sub.nR with group R as defined above, OCH.sub.2(NHCH.sub.2CH.sub.2).sub.nOCH.sub.3, OCH.sub.2(CH.sub.2).sub.nCO(NHCH.sub.2CH.sub.2).sub.nOCH.sub.3, OCH.sub.2(CH.sub.2).sub.nSO.sub.2(NHCH.sub.2CH.sub.2).sub.nOCH.sub.3, N(CH.sub.2CH.sub.2).sub.2N(CH.sub.2CH.sub.2).sub.nNH, CH.sub.3N(CH.sub.2CH.sub.2).sub.2N(CH.sub.2CH.sub.2).sub.n, CH.sub.3(CH.sub.2).sub.nCON(CH.sub.2CH.sub.2).sub.2N(CH.sub.2CH.sub.2).sub.n, CH.sub.3(CH.sub.2).sub.nSO.sub.2N(CH.sub.2CH.sub.2).sub.2N(CH.sub.2CH.sub.2).sub.n, O(CH.sub.2CH.sub.2).sub.2N(CH.sub.2CH.sub.2).sub.n, a monocyclic or bicyclic aryl, or an aromatic or non-aromatic heterocyclic system selected from the meanings of R;

[0029] M is selected from O, CH.sub.2, CHR.sub.2, CCHR.sub.2, C(R.sub.2).sub.m, CHOH, CHOR.sub.2, CCHOR.sub.2, CO, CONR.sub.2, NH, NR.sub.2, NOH, NOR.sub.2, NR.sub.2CO, S, SO, S(O).sub.2, wherein R.sub.2 is independently selected from the meanings of R, or R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8;

[0030] M.sub.1 is independently selected from the meanings of M, as defined above, or is absent;

[0031] R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8 are equal or different and independently selected from the meanings of R, or R, R.sub.1;

[0032] P is selected from O, Q, CH.sub.2, CHR.sub.2, C(R.sub.2).sub.m, CHOH, CHOR.sub.2, CHO-acyl, NH, N-acyl, NR.sub.2, NOH, NOR.sub.2 and NO-acyl, wherein R.sub.2 is independently selected from the meanings of R, or R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8;

[0033] P.sub.1 is independently selected from the meanings of P, as defined above, or is absent;

[0034] Q is selected from H, R.sub.2, OR.sub.2, O-acyl, NH.sub.2, NHR.sub.2, N(R.sub.2).sub.m, NH-acyl, NHCOOCH.sub.2Bn, NHCOO-t-Bu, NHCOOR, NHOH, NHOR.sub.2, N(R.sub.2)OR.sub.2 and NHO-acyl, wherein R.sub.2 is independently selected from the meanings of R, or R, R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8; or Q is a beta-lactam structure of formula (IX), (IX), (IX):

##STR00008##

[0035] wherein Q is selected from H, R.sub.2, OR.sub.2, O-acyl, NH.sub.2, NHR.sub.2, N(R.sub.2).sub.m, NH-acyl, NHCOOCH.sub.2Bn, NHCOO-t-Bu, NHCOOR, NHOH, NHOR.sub.2, N(R.sub.2)OR.sub.2 and NHO-acyl, wherein R.sub.2 is independently selected from the meanings of R, or R, R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8;

[0036] Q.sub.1 is independently selected from the meanings of Q, as defined above, or is absent;

[0037] T is a cyclic ring which is saturated or contains unsaturations and consists of 3 to 8 atoms bonded to one another and containing carbon atoms and/or N, O or S(O).sub.m, atoms, or substituted as in the meanings by R, R, R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8 or by X, M, P, W o Z or as explained in the meanings of the cyclic systems of G for R and/or R.sub.1;

[0038] T.sub.1 is independently selected from the meanings of T, as defined above, or is absent;

[0039] X is selected from the CH.sub.2, CH(R), C(R), O, S(O).sub.m, N, N(R), C(O), C(P), N(R)CO, CON(R) groups, wherein R is H or can represent one of the meanings defined for R, R and R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8;

[0040] W is selected from H, or the meanings of T or of Z or of R, or also of R, R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8;

[0041] Z is selected from H, CH.sub.3, CH.sub.2OCOCH.sub.3 or, taken together with the carbon atom to which it is bonded, Z is selected from CH.sub.2, CH(R.sub.2), C(O), C(P); or Z is selected from the meanings of R, R, R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7 and R.sub.8, or the meanings of M, P.

[0042] According to one embodiment, the compounds of general formula (I) are characterised by a substructure of formula (X), or by a substructure deriving from the substructure of formula (X), having formula (XI) or (XII):

##STR00009##

[0043] wherein

##STR00010##

is a substituent of formula IX, IX or IX

[0044] wherein m, X, R.sub.1, R.sub.2 and R.sub.3 are as defined above.

[0045] According to a further embodiment, the compounds of formula (XII) can have formula (XIII), (XIV) or (XV):

##STR00011##

[0046] wherein m, X, R.sub.1 and R.sub.3 are as defined above.

[0047] According to a preferred aspect of the invention, the compounds of formula (X) can have a formula (Xa-Xt), as reported below:

##STR00012## ##STR00013## ##STR00014##

[0048] or (Xa-Xm), as reported below:

##STR00015## ##STR00016##

[0049] wherein m, P, R.sub.1, R.sub.2 and R.sub.3 are as defined above.

[0050] According to a further preferred aspect of the invention, the compounds of formula (XI) can have formula (XIa-XIt) or (XIa-XIm), as reported below:

##STR00017## ##STR00018## ##STR00019## ##STR00020## ##STR00021##

[0051] wherein m, P, R.sub.1 and R.sub.3 are as defined above.

[0052] If Q is an azetidinone nucleus not substituted with the T ring as in formula (XIII), there can be structures of formula (XIIIa-XIIIt), (XIIIa-XIIIm):

##STR00022##

##STR00023## ##STR00024## ##STR00025##

##STR00026##

##STR00027## ##STR00028## ##STR00029##

[0053] wherein m, Q, R.sub.1 and R.sub.3 are as defined above.

[0054] If Q is a bicyclic system containing the beta-lactam nucleus 7-aminocefem substituted as in formula (XIV), there can be structures of formula (XIVa-XIVt), (XIVa, XIVm):

##STR00030##

##STR00031## ##STR00032## ##STR00033##

##STR00034##

##STR00035## ##STR00036## ##STR00037##

[0055] wherein m, P, Q, R.sub.1 and R.sub.3 are as defined above.

[0056] If Q is a bicyclic system containing the beta-lactam nucleus 6-aminopenam substituted as in formula (XV), there can be structures of formula XVa, XVb, XVc, XVh, XVm, XVn, XVo, XVp, XVq, XVr, XVs, XVt, XVa, XVb, XVc, XVd, XVe, XVf, XVg, XVh, XVi, XVl, XVm.

##STR00038## ##STR00039## ##STR00040##

##STR00041##

##STR00042## ##STR00043## ##STR00044##

[0057] wherein m, P, Q, R.sub.1 and R.sub.3 are as defined above.

[0058] According to a particularly preferred aspect of the invention, the compounds of general formula (I) are those listed in Table 1 below:

TABLE-US-00001 TABLE 1 # substructure # Code Structure XIc XIl RA1 [00045] XIa RA2 [00046] XIr RA3 [00047] Xa GA17M8 [00048] Xa GA17M10 [00049] XIb GA11S [00050] XIb RM66 [00051] XIb RM58 [00052] XIb RM70 [00053] XIb RM85 [00054] XId RA6 [00055] XId RA7 [00056] XId RA9 [00057] XId RA11 [00058] XId RA12 [00059] XId RA13 [00060] XIg RA14 [00061] Xc RA15 [00062] Xc RA16 [00063] XIc RA17 [00064] XIc RA18 [00065] XIc RA19 [00066] XIh RA20 [00067] Xe RA21 [00068] XIe RA22 [00069] XIVa GA09a [00070] XIVa GA09a [00071] XIVa GA09b [00072] XIVa GA09b [00073] XIVa RM36 [00074] XIVa RM37 [00075] XIVa RM53 [00076] XIVa RM47 [00077] XIVd RA23 [00078] XIVd RA24 [00079] XIVd RA25 [00080] XVe RA26 [00081] XVd RA27 [00082] XVd RA28 [00083] XVg RA29 [00084] XIIId RA30 [00085] XIVc RA31 [00086] XIVc RA32 [00087] XIVc RA33 [00088] XIIIc RA34 [00089] XVc RA35 [00090] XIVe RA36 [00091] XIVg RA37 [00092] XIVg RA38 [00093] XIVg RA39 [00094] XIVr RA40 [00095] XIVc RA41 [00096] XVr RA42 [00097] XIVc RA43 [00098] XIIIr RA44 [00099]

[0059] The condensed heterocycles A-D, substituted as in general formula (I), can be synthesised, depending on the type of heterocyclic system A-D or the need for substitution on the A-D system, with the various groupings R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8, M, M.sub.1, P, P.sub.1, T, T.sub.1, Q, X, W and Z, by means of:

[0060] 1condensation of ring A with the carbonyl system of the preformed heterocycle C-D with or without isolation of an intermediate hydrazone system (iia-iiq or iia-iii) and subsequent and/or simultaneous formation of the indole junction cycle B which gives rise to the desired heterocycle A-D. In this type of condensation, both aromatic systems are already substituted and/or substitutable with the various necessary groupings: R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8, M, M.sub.1, P, P.sub.1, T, T.sub.1, Q, X, W and Z, General Scheme (Ia).

##STR00100##

[0061] 2Condensation of a preformed indole system A-B already substituted with ring D to form junction ring C. In this type of condensation, as in 1-, both aromatic systems are already substituted and/or substitutable with the various necessary groupings: R, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8, M, M.sub.1, P, P.sub.1, T, T.sub.1, Q, X, W and Z, General Scheme (Ib).

##STR00101##

[0062] 3Condensation of a heterocyclic system substituted and/or substitutable as above containing the nucleus of the desired heterocyclic C-D which is activated as enamide and substituted with a Y substituted aryl (ring A is substituted and/or substitutable as above) [boronic acid B(OH).sub.2 or Si(Me).sub.3 or a mesityl-aryl-iodonium salt] to give an appropriate intermediate from which it will be cyclised with specific reagents to give indole junction ring B in the formation of heterocyclic system A-D substituted as desired for (I) and previously reported, General Scheme (Ic).

##STR00102##

[0063] Many of the condensation methods described above to obtain heterocyclic intermediates A-B and/or C-D and the reactions of formation of the desired tetracyclic systems A-D from said systems are also reported in Chem. Rev. 2002, 102, 4303-4427, Eur. J. Org. Chem. 2006, 1379-1382, J. Heterocyclic Chem., 48, 1095 (2011), Adv. Synth. Catal. 2010, 352, 363-367, Chem. Eur. J. 2010, 16, 1124-1127, J. Am. Chem. Soc. 2006, 128, 581-590, J. Org. Chem. 2003, 68, 2807-2811, J. Org. Chem. 2014, 79, 7836-7843, Org. Lett., 2012, 14, 14, 3772-3775.

[0064] For example, the tetracyclic heterocycles A-D of structure (Xa-Xt), General Schemes (Id-Il), were obtained by following said methods.

##STR00103##

[0065] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: an appropriately synthesised benzocondensed heptanone: NaH, THF or dioxane or another appropriate solvent, 3-OMe-benzaldehyde, THF or dioxane or an appropriate solvent and DMF; H.sub.2/PdC 10% EtOH, or alternatively an appropriate commercial R.sub.2-substituted-6,7,8,9-tetrahydro-benzocyclohepten-5-one; iii: 48% HBr, AcOH, 120 C., 12 h, or BBr.sub.3, dichloromethane, 78 C. to 0; iv: BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; v: 4-R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions (36% HCl reflux or AcOH, 120 C., 2-6 h, or microwave use in appropriate conditions 2-10 min., or BBr.sub.3, dichloromethane, 78 C. to 0 C., 2 hours; vi: appropriate R.sub.3X halide (mainly but not only Br or Cl), NaH/anh. DMF, 80 C., 16 h, or microwaves in appropriate conditions 2-10 min.; vii: KOH/EtOH.

##STR00104##

[0066] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i.sup.(1): a) 4-bromoethylbutyrate, KI, K.sub.2CO.sub.3, acetone, b) NaOH, EtOH; c) BnOH, K.sub.2CO.sub.3, DMF.sup.(1); ii.sup.(1): cyanuric chloride (C.sub.3N.sub.3Cl.sub.3), dichloromethane, pyridine, 0 C., 30+30 min, r/t 3 hours, cooled to 60 C., followed by addition of solid AlCl.sub.3 in portions, controlled t to 0 C. for 3-4 hours; iii: H.sub.2/PdC 10% EtOH or AcOEt/EtOH; iv: BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; v: 4-R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions (36% HCl reflux or AcOH, 120 C., 2-6 h, or microwave use in appropriate conditions 2-10 min.; vi: appropriate R.sub.3X halide (mainly but not only Br or Cl), NaH/anh. DMF, 80 C., 16 h, or microwaves in appropriate conditions 2-10 min.; vii: KOH/EtOH; [.sup.(1) European Journal of Organic Chemistry (2014), 2014, (15), 3170-3181; Org. Lett., Vol. 10, No. 13, 2008.].

##STR00105##

[0067] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: a) 4-bromoethylbutyrate, KOH, EtOH, N.sub.2 10 days or microwaves 1 h, H.sub.2O 5 hours, acidification pH=1 with 1N HCl; ii.sup.(i): trichloroacetic anhydride, 70 C., until complete (TLC); iii: HBr 48%, AcOH, 120 C., 12 h., or BBr.sub.3, dichloromethane, 78 C. to 0; iv: BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; v: 4-R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions (36% HCl reflux or AcOH, 120 C., 2-6 h, or microwave use in appropriate conditions 2-10 min, 2 hours; vi: appropriate R.sub.3X halide (mainly but not only Br or Cl), NaH/anh. DMF, 80 C., 16 h, or microwaves in appropriate conditions 2-10 min.; vii: KOH/EtOH; [.sup.(1) Synthetic Communications 1, 39: 2664-2673, 2009.].

##STR00106##

[0068] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: HBr 48%, AcOH, 120 C., 12 h., or BBr.sub.3, dichloromethane, 78 C. to 0 C.; MCPB, DCM r/t 2-6 hours or Ti(O-.sub.iPr).sub.4, (R,R)-diethyltartrate, DCM, 15 C., 15-30 min., or Oxone, THF/MeOH, r/t, 24-120 hours; iii: appropriate R.sub.3X halide (mainly but not only Br or CO, NaH/anh. DMF, 80 C., 16 h, or microwaves in appropriate conditions 2-10 iv: BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; iv: KOH/EtOH.

##STR00107##

[0069] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: pyridine, DCM, 4 hours r/t; K.sub.2CO.sub.3, MeCOEt, 12 hours r/t; t-BuOK, toluene, 12-18 hours r/t, then 6N HCl, AcOH, r/t, 12-18 h; iv: R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions 36% HCl reflux or AcOH, 120 C., 2-6 h, or microwave use in appropriate conditions 2-10 min, 2 hours; v: polyphosphoric acid, 50-80 C., inert atmosphere (Ar or N.sub.2), 1-2 hours, then NaOH/H.sub.2O to pH 8-9, extraction; vi: HBr 48%, AcOH, 120 C., 12 h., or BBr.sub.3, dichloromethane, 78 C. to 0; vii: BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; iv: KOH/EtOH; viii: KOH/EtOH.

##STR00108##

[0070] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: NMM, DMAP (cat.), DCM or THF, 0-20 C., 4 hours, or Et.sub.3N, H.sub.2O, dioxane; K.sub.2CO.sub.3, DMF or CH.sub.3CN, 12 hours r/t, KOH, EtOH, 12 hours, 10% HCl extraction; iii: t-BuOK, toluene, 12-18 hours r/t, then 6N HCl, AcOH, r/t, 12-18 h; iv: R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions HCl 36% reflux or AcOH, 120 C., 2-6 h, or microwave use in appropriate conditions 2-10 min, 2 hours; v: polyphosphoric acid, 50-80 C., inert atmosphere (Ar or N.sub.2), 1-2 hours, then NaOH/H.sub.2O to pH 8-9, extraction; vi: HBr 48%, AcOH, 120 C., 12 h., or BBr.sub.3, dichloromethane, 78 C. to 0; vii: BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; iv: KOH/EtOH; viii: KOH/EtOH.

##STR00109##

[0071] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: CsCO.sub.3, DMF, 1 hour, r/t; ii: irradiation at >250 in CH.sub.3CN, [Tetrahedron Lett., 34, 37, 1993, 5855-58]; iii: R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions 36% HCl reflux or AcOH, 120 C., 2-6 h, or microwave use in appropriate conditions 2-10 min, 2 hours; iv: HBr 48%, AcOH, 120 C., 12 h., or BBr.sub.3, dichloromethane, 78 C. to 0; v: BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; vi: KOH/EtOH.

##STR00110##

[0072] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions 36% HCl reflux or AcOH, 120 C., from 2-6 h to 5 days, or microwave use in appropriate conditions 2-10 min, 2 hours, [(+)* according to conditions X and XI a and b mixed or a alone]; ii: AcOH, Pd/C 120 C., 20 hours or iia: (2,2,6,6-tetramethyl-piperidin-1-yl)oxyl (TEMPO)/HBF.sub.4 50%, 0 C., 10 min, iib: N-oxoammonium-TEMPO, CH.sub.3CN, 0 C., 15 min; iii: appropriate R.sub.3X halide (mainly but not only Br or CO, NaH/anh. DMF, 80 C., 16 h, or microwaves in appropriate conditions 2-10 min.; iv: HBr 48%, AcOH, 120 C., 12 h, or BBr.sub.3, dichloromethane, 78 C. to 0; v: BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; vi: KOH/abs. EtOH, r/t, 20 hours/H.sup.+.

##STR00111##

[0073] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: 3-Cl-propionic acid, 20% KOH, 100 C., 3 days; ii: Polyphosphoric acid, 70 C., 1 hour; iii: R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions 36% HCl reflux or AcOH, 120 C., from 2-6 h to 5 days, or polyphosphoric acid, 70-75 C., 1.5 hours, then 2 hours r/t, or microwave use in appropriate conditions 2-10 min, 2 hours; iv: appropriate R.sub.3X halide (mainly but not only Br or Cl), NaH/anh. DMF, 80 C., 16 h, or microwaves in appropriate conditions 2-10 min.; v: HBr 48%, AcOH, 120 C., 12 h, or BBr.sub.3, dichloromethane, 78 C. to 0; v: pyridine hydrochloride 180-190 C., inert atmosphere (N.sub.2 or Ar) 2 hours, or pyridine hydrochloride microwaves 5 min; vi: BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; vii: KOH/abs. EtOH, r/t, 20 hours/H.sup.+.

##STR00112##

[0074] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: 3-Cl-propionic acid, 20% NaOH, 100 C. for 2 hours then 60 C. for 12 hours, r/t/HCl 3 hours and extraction; ii: Polyphosphoric acid, 50 C., 4 hours; iii: R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions 36% HCl reflux or AcOH, 120 C., from 2-6 h to 5 days, or microwave use in appropriate conditions 2-10 min, 2 hours; iv: appropriate R.sub.3X halide (mainly but not only Br or Cl), NaH/anh. DMF, 80 C., 16 h, or microwaves in appropriate conditions 2-10 min.; v: HBr 48%, AcOH, 120 C., 12 h, or BBr.sub.3, dichloromethane, 78 C. to 0; v: pyridine hydrochloride 180-190 C., inert atmosphere (N.sub.2 or Ar) 2 hours, or pyridine hydrochloride microwaves 5 min; vi: BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; vii: KOH/abs. EtOH, r/t, 20 hours/H.sup.+.

##STR00113##

[0075] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions 36% HCl reflux or AcOH, 120 C., from 2-6 h to 5 days, or microwave use in appropriate conditions 2-10 min, 2 hours; ii: MCPB, DCM. r/t 2-6 hours or Ti(O-.sub.iPr).sub.4, (R,R)-diethyltartrate, DCM, 15 C., 15-30 min. or Oxone, THF/MeOH, r/t, 24-120 hours; iii: appropriate R.sub.3X halide (mainly but not only Br or Cl), NaH/anh. DMF, 80 C., 16 h, or microwaves in appropriate conditions 2-10 min.; iv: HBr 48%, AcOH, 120 C., 12 h, or BBr.sub.3, dichloromethane, 78 C. to 0; iv: pyridine hydrochloride 180-190 C., inert atmosphere (N.sub.2 or Ar) 2 hours, or pyridine hydrochloride microwaves 5 min; v: BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; vi: KOH/abs. EtOH, r/t, 20 hours/H.sup.+.

##STR00114##

[0076] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions 36% HCl reflux or AcOH, 120 C., from 2-6 h to 5 days, or microwave use in appropriate conditions 2-10 min, 2 hours; ii: appropriate R.sub.3X halide (mainly but not only Br or Cl), NaH/anh. DMF, 80 C., 16 h, or microwaves in appropriate conditions 2-10 min.; HBr 48%, AcOH, 120 C., 12 h, or BBr.sub.3, dichloromethane, 78 C. to 0; iii: pyridine hydrochloride 180-190 C., inert atmosphere (N.sub.2 or Ar) 2 hours, or pyridine hydrochloride microwaves 5 min; iv: BrCH.sub.2COOEt, KOH, EtOH 85%, r/t, 2 h, or BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone or DMF, 70 C.; v: KOH/abs. EtOH, r/t, 20 hours/H.sup.+.

[0077] The intermediate 7-methoxy-1,1-dioxo-3,4-dihydro-2H-1,2-benzothiazin-4-one is prepared according to Scheme (Io) below.

##STR00115##

[0078] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: NaH, DMF, 30 min r/t, methyl-chloroacetate 110 C.; ii: NaOMe, MeOH, 55 C., 30 min, HCl, 5 C. pH=3; iii: HCl/H.sub.2O reflux.

##STR00116##

[0079] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions 36% HCl reflux or AcOH, 120 C., from 2-6 h to 5 days, or microwave use in appropriate conditions 2-10 min, 2 hours; ii: appropriate R.sub.3X halide (mainly but not only Br or Cl), NaH/anh. DMF, 80 C., 16 h, or microwaves in appropriate conditions 2-10 min.; HBr 48%, AcOH, 120 C., 12 h, or BBr.sub.3, dichloromethane, 78 C. to 0; iii: pyridine hydrochloride 180-190 C., inert atmosphere (N.sub.2 or Ar) 2 hours, or pyridine hydrochloride microwaves 5 min; iv: BrCH.sub.2COOEt, KOH, EtOH 85%, r/t, 2 h or BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone or DMF, 70 C.; v: KOH/abs. EtOH, r/t, 20 hours/H.sup.+.

[0080] The intermediate 7-methoxy-2,3-dihydro-isoquinolin-1,4-dione is prepared according to the following scheme (Scheme Ip).

##STR00117##

[0081] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: NaH, DMF, 30 min r/t, methyl-chloroacetate 110 C.; ii: NaOMe, MeOH, 55 C., 30 min, HCl, 5 C. pH=3; iii: H.sub.2SO.sub.4/H.sub.2O reflux, 1 hour.

##STR00118##

[0082] General reaction conditions for oxidation of methylenes (Xa and Xa) aryl-conjugated to carbonyl derivatives of type Xm and Xg for the synthesis of derivatives of formulas: XIm, XIg, XIIm, XIIg, XIIIm, XIIIg, XIVm, XIVg-reagents, solvents and appropriate conditions according to type of reaction: i: NaBH.sub.4, Bi.sub.2O.sub.3, distilled H.sub.2O r/t, forms a black precipitate washed with H.sub.2O; ii: pyridine, AcOH, picolinic acid and t-BuOOH in H.sub.2O (70). 30 min microwave heating at 100 C. for 16 h (glass vial), cool, dilute with DCM, filter through celite, evaporate at 1/p [Org. Lett., 7, 21, 4549-4552, 2005 and references cited therein].

##STR00119##

[0083] Reaction conditionsreagents, solvents and appropriate conditions according to type of reaction: i: [precursor synthesised as shown in JOC 67, 10, 3502-3505, 2002], polyphosphoric acid, 100 C., K.sub.2CO.sub.3/H.sub.2O neutralisation, or propyl phosphonic acid, 90 C., 20 min, neutralisation and extraction; ii: Acetic anhydride or trifluoro acetic anhydride, 100 C., 16 hours, or acetyl chloride, toluene, pyridine, r/t, 12 hours; iii: R.sub.1-phenylhydrazine, or 4-R.sub.1-phenylhydrazine hydrochloride or hydrobromide and appropriate acidic conditions 36% HCl reflux or AcOH, 120 C., from 2-6 h to 5 days, or microwave use in appropriate conditions 2-10 min, 2 hours; iv: appropriate R.sub.3X halide (mainly but not only Br or Cl), NaH/anh. DMF, 80 C., 16 h, or microwaves in appropriate conditions 2-10 min.; v: BBr.sub.3, dichloromethane, 78 C. to 0, or: pyridine hydrochloride reflux 5 min, inert atmosphere (N.sub.2 or Ar), H.sub.2O, or pyridine hydrochloride microwaves 5 min, H.sub.2O; vi: BrCH.sub.2COOEt, KOH, EtOH 85%, r/t, 2 h or BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone or DMF, 70 C.; vii: DBU, MeOH or CH.sub.3CN, microwaves 20 min-4 hours or reflux 2-4 hours [Synth. Commun., 32(2), 265-272 (2002)]; or H.sub.2SO.sub.4/H.sub.2O, MeOH or THF, reflux 5 min, then K.sub.2CO.sub.3/H.sub.2O, r/t [Org. Lett. 16, 19, 5156-5159, 2014]; viii: appropriate R.sub.1X halide (mainly but not only Br or Cl), NaH/anh. DMF, 80 C., 16 h, or microwaves in appropriate conditions 2-10 min.; ix: KOH/abs. EtOH, r/t, 20 hours/H.sup.+.

[0084] The structures (X and X) substituted in R.sub.2 on the D ring as in (XI and XI), which can preferably be acids (Q=OH), esters (Q=OR.sub.1) or amides (Q=NHR.sub.1), or in another preferential embodiment amides with a Q group consisting of a generic beta-lactam system as in general substructure XII from which amides with 3-amino-4-oxo azetidinones (XIII and XIII) derive, or as in the case of amides with 7-amino-cephalosporin nuclei (XIV and XIV), or as in the case of 6-amino-penicillin nuclei (XV and XV), are obtainable either directly or by appropriate condensation according to General Scheme II:

##STR00120##

[0085] Reaction conditionsis Lvg=Halogen (Cl or Br or I) or MsO or TsO ester or OH; appropriate reagents, solvents and conditions according to the type of reaction; ii: (appropriate halide, Cl or Br or I), appropriate base (KOH or NaOH or K.sub.2CO.sub.3 or CsCO.sub.3, appropriate solvents and conditions); iii: KOH (appropriate solvents and conditions); iv: acid activation: a) C.sub.2O.sub.2Cl.sub.2, DMFcat, (solvents and appropriate conditions to form the suitable acid chloride of XI and XI), or a) ClCOOR, (solvents and appropriate conditions to form the suitable mixed anhydride of XI and XI), or a) activation as N-hydroxysuccinyl-ester, (solvents and appropriate conditions to form the suitable NOS ester of XI and XI), or a) ECDI or other appropriate dehydrating agents (solvents and appropriate condensation conditions).

[0086] A further subject of the present invention is the use of the compounds of general formula (I) as described above for the treatment and diagnosis of degenerative disorders characterised by high cell proliferation and/or tissue degeneration.

[0087] The compounds of the invention can be used as therapeutic agents or diagnostic probes in degenerative disorders which may be endogenous or exogenous (ie. induced by infectious agents such as bacteria, viruses, protozoa or fungi), characterised by high cell proliferation and/or tissue degeneration.

[0088] The disorders which can be treated or diagnosed with the compounds of the invention are cancers of various kinds, and other disorders such as infectious disorders caused by pathogens wherein physiological homeostatic tissue control has been lost, and control of proliferative activity and cell apoptosis is important.

[0089] The following examples further illustrate the invention.

EXAMPLES

[0090] Materials and Methods

[0091] The .sup.1H-NMR spectra were determined with a Varian Gemini 200 spectrometer operating at 200 MHz, or a Bruker Advance III HD 400 operating at 400 MHz. Chemical shifts (6) are expressed in parts per million (ppm). Coupling constants J are reported in Hertz. The following abbreviations have been used: singlet (s), doublet (d), triplet (t), broad singlet (br s) and multiplet (m). The preparatory Liquid Chromatography was conducted with flash chromatography, using pre-packed Isolute columns (Biotage) and glass columns containing silica gel 230-400 mesh. Thin-Layer Chromatography (TLC) was conducted with 60 F254 (MERCK) silica gel plates containing a fluorescent indicator. The various spots were highlighted with a UV lamp (256 nM). Evaporation was conducted under vacuum in a rotary evaporator, using anhydrous Na.sub.2SO.sub.4 as dehydrating agent.

[0092] The compounds of general formulas Xa to XVm can be prepared, for example, as reported below for (XIVa # RM37) and for the corresponding acid (XIVa # RM53) (Scheme IIa).

Example 1Preparation of (6R, 7R)-tert-butyl-7-(2-(8-fluoro-6,11-dihydro-5H-benzo[a]carbazol-3-yloxy)acetamido)-3-(acetoxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate (XIVa # RM37) and its Corresponding Acid (6R,7R)-7-(2-[(8-fluoro-6,11-dihydro-5H-benzo[a]carbazol-3-yl)oxy]acetamido)-3-(acetoxymethyl)-8-oxy-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic (XIVa # RM53), Scheme IIa

[0093] ##STR00121##

[0094] Reagents and conditions: a) HBr 48%, AcOH, 120 C., 12 h; b) BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; c) AcOH, 120 C., 6 h; d) KOH/EtOH; e) isobutene, H.sub.2SO.sub.4, dioxane; f) EDC, THF, 0 C.; g) TFA, anisole, CH.sub.2Cl.sub.2, 0 C.

Synthesis of 7-hydroxy-3,4-dihydronaphthalen-1(2H)-one (3)

[0095] An aqueous solution of 48% HBr (62 mL) was added to a solution of 6-methoxy-1-tetralone (6 g, 34.05 mmols) in glacial AcOH (9 mL), and the reaction mixture was left at reflux at 120 C. under stirring overnight. The reaction was cooled and then evaporated under vacuum to give the crude compound 3 as a black semisolid. The residue was then purified by crushing with n-hexane to give 3 as a brown solid (6.76 g, 98% yield). .sup.1H-NMR (Acetone-d.sub.6) : 1.98-2.18 (m, 4H); 2.80-2.95 (m, 2H); 6.70-6.82 (m, 2H); 7.80-7.90 (m, 1H).

Synthesis of 2-(1,2,3,4-tetrahydro-1-oxonaphthalen-7-yloxy) ethyl acetate (4)

[0096] Potassium carbonate (17.120 g, 125.11 mmols) was added to a solution of hydroxy tetralone 3 (6.76 g, 41.70 mmols) in anhydrous acetone (200 mL) placed under nitrogen, and the mixture was left under stirring for 1 hour at room temperature. Ethyl bromoacetate (7 mL) was then added, and the reaction was left under stirring overnight at 57 C., then filtered to eliminate inorganic salts, and finally, the filtrate was evaporated at low pressure. The resulting residue was then taken up with AcOEt (200 mL) and washed with a saturated solution of NaHCO.sub.3(2100 mL) and brine (2100 mL). The organic phase was then dried and evaporated to give a brown oil. Finally, the crude product was purified by flash chromatography on silica gel (n-hexane/AcOEt 5:1) to give the pure compound 4 as a yellow oil (6.67 g, 61% yield). .sup.1H-NMR (CDCl.sub.3) : 1.30 (t, J=7.1 Hz, 3H); 2.05-2.17 (m, 2H); 2.61 (t, J=6.8 Hz, 2H); 2.92 (t, J=6.0 Hz, 2H); 4.3 (q, J=7.1 Hz, 2H); 4.67 (s, 2H); 6.71-6.84 (m, 2H); 8.0 (d, J=8.6 Hz, 1H).

Synthesis of ethyl 2-[(8-fluoro-6,11-dihydro-5H-benzo[a]carbazol-3-yl)oxy]acetate (5)

[0097] A mixture of tetralone 4 (1.0 g, 3.80 mmols) and (4-fluorophenyl)hydrazine (0.643 g, 3.95 mmols) in glacial AcOH (5.3 mL) was placed at reflux at 120 C. for 6 hours under an inert atmosphere. The suspension was then cooled to room temperature and evaporated to give a solid yellow residue. The residue was taken up with AcOEt (150 mL), washed with H.sub.2O (680 mL) and saturated solution of NaHCO.sub.3 (180 mL), dried on Na.sub.2SO.sub.4 and evaporated at low pressure. The crude product obtained was then purified by crushing with n-hexane/Et.sub.2O to give 5 as a yellow solid (0.920 g, 72% yield). .sup.1H-NMR (CDCl.sub.3) : 1.32 (t, J=7.2 Hz, 3H); 2.85-3.09 (m, 4H); 4.22 (q, J=7.2 Hz, 2H); 4.65 (s, 2H); 6.72-6.72 (m, 6H); 7.08-7.31 (m, 3H); 8.18 (brs, 1H).

Synthesis of 2-[(8-fluoro-6,11-dihydro-5H-benzo[a]carbazol-3-yl)oxy]acetic acid (6)

[0098] A mixture of carbazole 5 (0.920 g, 2.71 mmols) and KOH (0.182 g, 3.25 mmols) in absolute EtOH (65 mL) was left under stirring at room temperature for 2 days. The solvent was then evaporated and the resulting yellow residue was dissolved in H.sub.2O. The aqueous solution was acidified with 10% HCl to pH 1 and extracted with AcOEt (4100 mL). The organic phase, dried and evaporated, supplied carboxylic acid 6 as a yellow solid (0.767 g, 91% yield). .sup.1H-NMR (DMSO-d.sub.6) : 2.75-2.90 (m, 2H); 2.92-3.05 (m, 2H); 4.70 (s, 2H); 6.79-6.84 (m, 1H); 6.86-6.96 (m, 2H); 7.14-7.24 (m, 1H); 7.26-7.38 (m, 1H); 7.51-7.63 (m, 1H); 11.42 (s, 1H).

Synthesis of (6R, 7R)-tert-butyl-3-(acetoxymethyl)-7-amino-8-oxo-5-thia-1-aza-bicyclo[4.2.0]oct-2-ene-carboxylate (7)

[0099] Isobutene (32 mL) was added to a mixture of 7-aminocephalosporanic acid (7-ACA) (7.0 g, 25.71 mmols) and conc. H.sub.2SO.sub.4 (6.42 mL) in anhydrous dioxane (64 mL), placed at 0 C. The reaction mixture was stirred at room temperature for 2 hours. The mixture was then adjusted to an alkaline pH by adding a saturated solution of NaHCO.sub.3, extracted with AcOEt (3200 mL), washed with brine (5100 mL), dried and evaporated to give a brown oil. The crude product was then purified by crushing with n-hexane to give pure 7 as a yellow solid (4.154 g, 50% yield). .sup.1H-NMR (CDCl.sub.3) : 1.56 (s, 9H); 2.11 (s, 3H); 2.02-2.05 (brs, 2H); 3.40-3.57 (2d, J=18.2 Hz, 2H); 4.78-4.85 (m, 2H); 5.0 (d, J=5.1 Hz, 1H); 5.07 (d, J=13.2 Hz, 1H).

Synthesis of (6R, 7R)-tert-butyl-7-(2-(8-fluoro-6,11-dihydro-5H-benzo[a]carbazol-3-yloxy)acetamido)-3-(acetoxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate (XIVa; # RA/137)

[0100] 7-ACA tert-butyl ester 7 (0.211 g, 0.642 mmols) was added to a solution of carboxylic acid 6 (0.200 g, 0.642 mmols), in anhydrous THF (31 mL). The reaction mixture was cooled in an ice bath, and EDC (0.136 g, 0.642 mmols) was added gradually. The reaction was left under stirring in an inert atmosphere at room temperature for 24 hours. The solvent was evaporated at room temperature and the residue was taken up with CHCl.sub.3 (60 mL), washed with H.sub.2O (425 mL), dried on Na.sub.2SO.sub.4 and evaporated to give a yellow solid. The crude product was purified by flash chromatography (n-hexane/AcOEt 3:1) using an Isolute column (Si II) to give XIVa as a yellow oil (0.267 g, 67% yield). .sup.1H-NMR (CDCl.sub.3) : 1.55 (s, 9H); 2.07 (s, 3H); 2.87-2.94 (m, 2H); 2.99-3.06 (m, 2H); 3.31-3.58 (2d, J=18.4 Hz, 2H); 4.58 (s, 2H); 4.76-4.83 (m, 1H); 4.99-5.05 (m, 2H); 5.89-5.94 (q, J=4.9 Hz, 1H); 6.76-6.81 (m, 1H), 6.85-6.92 (m, 2H); 7.12-7.17 (m, 1H); 7.24-7.30 (m, 2H); 8.30 (s, 1H).

Synthesis of (6R,7R)-7-(2-[(8-fluoro-6,11-dihydro-5H-benzo[a]carbazol-3-yl)oxy]acetamido)-3-(acetoxymethyl)-8-oxy-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (XIVa, # RM53)

[0101] Anisole (0.05 mL) and TFA (0.42 mL) were added to a solution of compound 1 (60 mg, 0,097 mmols) in anhydrous CH.sub.2Cl.sub.2 (1.0 mL) placed on ice. The reaction mixture was stirred at room temperature for 18 hours under a nitrogen atmosphere. The reaction was then evaporated at room temperature, and the resulting yellow solid was crushed with Et.sub.2O to give compound XIVa as a yellow solid (18 mg, 33% yield). .sup.1H-NMR (DMSO-d.sub.6) : 2.04 (s, 3H); 2.81-2.88 (m, 2H); 2.94-3.0 (m, 2H); 3.48-3.69 (2d, J=18 Hz, 2H); 4.65-4.74 (m, 2H); 5.0 (d, J=12.8 Hz, 1H); 5.15 (d, J=4.8 Hz, 1H); 5.73-5.77 (q, J=4.8 Hz, 1H); 6.84-6.94 (m, 3H), 6.78-7.23 (m, 1H); 7.29-7.34 (m, 1H); 7.55-7.60 (m, 1H); 9.13 (d, J=8.4 Hz, 1H); 11.40 (s, 1H).

[0102] Example 2by analogy with XIVa; # RM37, compounds XIVa (# GA09a, # GA09b, RM36, RM47) can be prepared, as reported below, and consequently, as for acid XIVa # RM53, their corresponding acids of type XIVa (# GA09a, # GA09b9) can be obtained (Scheme II b)

##STR00122##

[0103] Reagents and conditions: a) HBr 48%, AcOH, 120 C., 12 h; b) BrCH.sub.2COOEt, K.sub.2CO.sub.3, acetone, reflux; c) AcOH, 120 C., 6 h; d) KOH/EtOH; e) isobutene, H.sub.2SO.sub.4, dioxane; f) EDC, THF, 0 C.; g) TFA, anisole, CH.sub.2Cl.sub.2, 0 C.

Example of Preparation of (6R, 7R)-tert-butyl-7-(2-(7-nitro-6,11-dihydro-5H-benzo[a]carbazol-3-yloxy)acetamido)-3-(acetoxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate (XIVa # GA09a), (6R, 7R)-tert-butyl-7-(2-(9-nitro-6,11-dihydro-5H-benzo[a]carbazol-3-yloxy)acetamido)-3-(acetoxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate (XIVa # GA09b) and their Corresponding Acids (6R,7R)-7-(2-[(7-nitro-6,11-dihydro-5H-benzo[a]carbazol-3-yl)oxy]acetamido)-3-(acetoxymethyl)-8-oxy-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (XIVa # GA09a) and (6R,7R)-7-(2-[(9-nitro-6,11-dihydro-5H-benzo[a]carbazol-3-yl)oxy]acetamido)-3-(acetoxymethyl)-8-oxy-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (XIVa # GA09b) Scheme IIb

Example 3General Method for the Preparation of 8-(R)-5,6-dihydrobenzo[a]carbazoles (XIa)

[0104] A suitable R-phenylhydrazine (e.g. R=4-MeO or 4-Cl or 4-Br or 3-NO.sub.2) (8.39 mmols) is added to a tetralone solution 4 (8.05 mmols) in glacial acetic acid (11.4 ml), and the mixture is maintained at reflux and under stirring for 8 hours. After said time has elapsed, the reaction is cooled in an ice bath to facilitate the formation of a precipitate. The crystalline solid is separated from the reaction mixture by vacuum filtration. The crystalline solid obtained is washed 4/5 times with water to eliminate the excess glacial acetic acid. The precipitation mother liquors (acetic acid) are evaporated at 1/p, and the resulting residue is crystallised several times from EtOH/Et.sub.2O and vacuum filtered.

[0105] XIa (R=8-MeO):

[0106] Yield: 17.21%. M.p.: 168-169. .sup.1HNMR (DMSO-d6) : 1.25 (t, J=7.2 Hz, 3H), 2.91 (m, 4H), 3.78 (s, 3H), 4.20 (q, J=7.2 Hz, 2H), 4.78 (s, 2H) 6.62-7.71 (m, 6H), 11.05 (s, H). MS m/e 352 (M+)

[0107] XIa (R=8-Cl):

[0108] Yield: 50.59%. M.p.: 147-148. .sup.1HNMR (DMSO-d6) : 1.25 (t, J=7.2 Hz, 3H), 2.92 (m, 4H), 4.20 (q, J=7.2 Hz, 2H), 4.79 (s; 2H), 6.81-7.78 (m, 6H), 11.45 (s, H). MS m/e 356 (M+)

[0109] XIa (R=8-Br):

[0110] Yield: 39.99%. M.p.: 154-155. .sup.1HNMR (DMSO-d6) : 1.25 (t, J=7.2 Hz, 3H), 2.91 (m, 4H), 4.20 (q, J=7.2 Hz, 2H), 4.79 (s, 2H) 6.81-7.71 (m, 6H), 11.47 (s, H). MS m/e 400 (M+)

Example 4General Method for the Preparation of 7-(R)-5,6-dihydrobenzo[a]carbazoles and 9-(R)-5,6-dihydrobenzo[a]carbazoles (XIa)

[0111] The appropriate 3-R phenyl hydrazine (e.g. R=3-NO.sub.2) (11.36 mmols) is added to a solution of the appropriate tetralone 3 (11.0 mmols) in glacial acetic acid (15.5 ml), and the mixture is heated to reflux under stirring for 6 hours. When that time has elapsed, the solvent (acetic acid) is removed by evaporation at 1/p and the semisolid residue obtained, dissolved in THF and preabsorbed on silica gel 60 (Merck 70-230 mesh, using a mixture/silica weight ratio of 1/7), is MPLC chromatographed on a silica gel 60 (Merck 230-400 mesh) column (d: 4 cm, h: 45 cm), using a hexane-ethyl acetate 2:1 mixture as eluent and collecting fractions of about 20 ml.

[0112] The two regioisomers 7-(NO.sub.2)-5,6-dihydrobenzo[a]carbazole (XIa, R=7-NO.sub.2) and 9-(NO.sub.2)-5,6-dihydrobenzo[a]carbazole (XIa, R=9-NO.sub.2) are separated by the chromatography (present in the crude reaction product at the ratio of 1:1 demonstrated by 1HMNR).

[0113] XIa (R=7-NO.sub.2):

[0114] Yield: 10.6% M.p.: 221-222 C. .sup.1HNMR (DMSO-d6): 1.24 (t, J=7.2 Hz, 3H), 2.85-3.15 (m, 2H), 4.19 (q, J=7.2 Hz, 2H), 4.80 (s, 2H), 6.50-8.60 (m, 6H), 11.52 (s, H).

[0115] XIa (R=9-NO.sub.2):

[0116] Yield: 29.7% M.p.: 179-180 C. .sup.1HNMR (DMSO-d6): 1.23 (t, J=7.2 Hz, 3H), 1.77-2.01 and 2.42-2.80 (2m, 4H), 4.18 (q, J=7.2 Hz, 2H), 4.77 (s, 2H) 6.74-6.88, 6.62-7.00 and 7.82-8.21 (3m, 6H), 10.05 (s, H).

Example 5

[0117] Similarly to the compounds of type Xa, benzyl esters 15a and 15c can be prepared as pure from tetralones 16-18; for example, 5,6-dihydrobenzo[a]carbazoles (XIa) 19-24 are prepared.

##STR00123##

[0118] Conditionsi: HBr, CH3CO2H, reflux, 12 h; RBr, acetone, K2CO3, reflux, 12 h; iii: 12 or 13, 3-NO2-PhNHNH2 or 4-NO2-PhNHNH2, CH3CO2H, reflux, 6 h; iv: 16-18, 3-NO2-PhNHNH2, CH3CO2H, reflux, 6 h.

Synthesis of benzyl 2-(1,2,3,4-tetrahydro-1-oxonaphthalen-7-yloxy) acetate (4)

[0119] The preparation is similar to that of the synthesis of ethyl 2-(1,2,3,4-tetrahydro-1-oxonaphthalen-7-yloxy) acetate (4). In this case, benzyl bromoacetate is used as reagent instead of ethyl bromoacetate. As in the case of 4, 4 also presents as a transparent oil. (93% yield). 1HNMR (CDCl3): 2.14 (m, 2H), 2.63 (t, J=5.6 Hz, 2H), 2.90 (t, J=6.4 Hz, 2H), 4.72 (s, 2H), 5.25 (s, 2H), 6.65-8.12 (m, 8H).

Synthesis of 5,6-dihydrobenzo[a]carbazoles (XIa) 15a and 15c (benzyl esters)

[0120] Similarly to the ethyl esters of 5,6-dihydrobenzo[a]carbazoles (XIa) previously described using tetralone 4, benzyl esters 15a and 15c were obtained.

[0121] 15a (R=7-NO.sub.2): Yield: 14.4%; m.p.: 185-186 C.; .sup.1HNMR (DMSO-d6): 2.85-3.15 (m, 4H), 4.91 (s, 2H), 5.21 (s, 2H), 6.90-7.85 (m, 11H), 12.25 (s, H).

[0122] 15c (R=9-NO.sub.2): Yield: 56.0%; m.p.: 205-206 C.; .sup.1HNMR (DMSO-d6): 2.85-3.05 (m, 4H), 4.91 (s, 2H), 5.21 (s, 2H), 6.85-8.45 (m, 11H), 12.18 (s, H).

Synthesis of 5,6-dihydrobenzo[a]carbazoles (Xa) 19-24

[0123] Using tetralones 16-18 as described in the synthesis of 5 with tetralone 3,6-dihydrobenzo[a]carbazoles (Xa) 19-24 were obtained.

[0124] 19: Yield: 73.2% .sup.1HNMR (DMSO-d6): 2.60-3.10 (m, 4H), 3.63 (s, 3H), 6.43-7.96 (m, 6H), 12.20 (s, H).

[0125] 20: Yield: 83% .sup.1HNMR (DMSO-d6): 2.51-2.76 (m, 4H), 3.50 (s, 3H), 6.03-7.53 (m, 6H), 12.20 (s, H).

[0126] 21: Yield: 75% .sup.1HNMR (DMSO-d6): 2.40-2.96 (m, 4H), 3.86 (s, 3H), 3.93 (s, 3H), 6.60-8.11 (m, 5H).

[0127] 22: Yield: 74% .sup.1HNMR (DMSO-d6): 2.58-3.11 (m, 4H), 3.61 (s, 3H), 6.44-7.90 (m, 6H), 12.21 (s, H).

[0128] 23: Yield: 78% .sup.1HNMR (DMSO-d6): 2.50-2.72 (m, 4H), 3.54 (s, 3H), 6.13-7.53 (m, 6H), 12.20 (s, H).

[0129] 24: Yield: 72% .sup.1HNMR (DMSO-d6): 2.38-2.89 (m, 4H), 3.82 (s, 3H), 3.73 (s, 3H), 6.48-8.01 (m, 5H).

Example 6Synthesis Scheme for 5,6-dihydrobenzo[a]carbazoles type Xa (39, 42, 43, 46, 49, 59-63, 67, 71, 72)

[0130] ##STR00124##

Synthesis of 5,6-dihydrobenzo[a]carbazoles Xa (39, 42, 59-61)

[0131] The appropriate 4-R-phenylhydrazine (4-NO.sub.2; 4-MeO; 4-Cl; 4-Br; 4-F) (59.01 mmols) is added to a solution of 6-methoxy-1-tetralone (56.74 mmols) in glacial acetic acid (80.32 ml) and the mixture is maintained at reflux, under stirring, for 8 hours. After said time, a crystalline solid is separated from the reaction mixture by cooling in an ice bath, vacuum filtered and washed several times with H.sub.2O to eliminate the excess glacial AcOH. The mother liquor (acetic acid), still containing aliquots of the desired carbazole (39, 42, 59-61) mixed with 6-methoxy-1-tetralone, is evaporated at 1/p, and the residue is crystallised several times with EtOH. The precipitate, purified by crystallisation, is vacuum filtered, supplying other pure aliquots of the corresponding carbazoles.

[0132] 39: Yield: 82%. M.p.: 170-171. .sup.1HNMR (CDCl.sub.3): 2.89-3.09 (m, 4H), 3.80 (s, 3H), 6.69-7.41 (m, 6H), 8.06 (s, H).

[0133] MS m/e 283 (M+)

[0134] 42: Yield: 33%. M.p.: 186-187. .sup.1HNMR (CDCl.sub.3): 2.91-3.00 (m, 4H), 3.78 (s, 3H), 3.88 (s, 3H), 6.61-8.42 (m, 6H), 11.03 (s, H). MS m/e 279 (M.sup.+)

[0135] 59: Yield: 10%. M.p.: 197-198. .sup.1HNMR (CDCl.sub.3): 2.55-2.90 (m, 4H), 3.78 (s, 3H), 6.68-8.15 (m, 6H), 10.20 (s, H).

[0136] 60: Yield: 89%. M.p.: 175-176. .sup.1HNMR (CDCl3): 2.89-3.00 (m, 4H), 3.81 (s, 3H), 6.69-7.28 (m, 6H), 8.06 (s, H).

[0137] MS m/e 267 (M.sup.+)

[0138] 61: Yield: 97%. M.p.: 166-167. .sup.1HNMR (CDCl.sub.3): 0.73-3.09 (m, 4H), 3.81 (s, 3H), 6.66-8.12 (m, 6H), 8.76 (s, H).

[0139] MS m/e 328 (M.sup.+)

Synthesis of 5,6-dihydrobenzo[a]carbazoles Xa (49, 52, 62, 67, 71)

[0140] HBr 48% (0.64 ml) is added to a solution of the appropriate carbazole (39, 42, 59, 60, 61) (0.35 mmols) in glacial acetic acid (0.09 ml). The mixture is maintained at reflux and under stirring at 120 C. for 18 hours. After said time, the solvent is evaporated at 1/p to obtain a solid consisting of (49, 52, 62, 67, 71).

[0141] 49: Yield: 82%. M.p.: 215-216. .sup.1HNMR (DMSO-d6): 2.51-2.87 (m, 4H), 6.63-7.51 (m, 6H), 11.39 (s, H).

[0142] MS m/e 269 (M.sup.+)

[0143] 52: Yield: 98%. M.p.: >330. .sup.1HNMR (DMSO-d6): 2.49-2.81 (m, 4H), 6.42-7.33 (m, 6H), 10.79 (s, H).

[0144] MS m/e 251 (M.sup.+)

[0145] 62: Yield: 96%. M.p.: 183-184. .sup.1HNMR (DMSO-d6): 2.51-2.86 (m, 4H), 6.53-7.31 (m, 6H), 11.21 (s, H).

[0146] MS m/e 253 (M.sup.+)

[0147] 67: Yield: 91%. M.p.: 293-294. .sup.1HNMR (DMSO-d6): 2.48-2.52 (m, 4H), 7.10-8.50 (m, 6H), 11.39 (s, H).

[0148] 71: Yield: 48%. M.p.>330. .sup.1HNMR (DMSO-d6): 2.33-2.56 (m, 4H), 6.43-8.52 (m, 6H), 11.90 (s, H).

General Method for N-alkylation of 5,6-dihydrobenzo[a]carbazoles Xa (39, 42, 59-61)

[0149] The appropriate carbazole to be alkylated (39, 42, 59-61) (3.5 mmols) is added in portions to a solution of 60% NaH (15.5 mmols) in anh. DMF (7 ml) over a period of 30 minutes. The appropriate R.sub.3X alkyl halide (7.04 mmols) [in the example R.sub.3X(N,N-dimethyl amino)-propyl chloride.HCl], dissolved in anh. DMF (5 ml), is then dripped in, and the mixture is left under stirring at 80 C. for 48 hours. The solvent is evaporated at 1/p, and the residue is taken up with MeOH HPLC, placed in an ice bath and salified with Et.sub.2O.HCl and anh. Et.sub.2O. The resulting precipitate (43, 46, 63, 72) is vacuum filtered.

[0150] 43: Yield: 95%. M.p.>330. .sup.1HNMR (DMSO-d6): 2.70-2.98 (m, 8H), 3.35 (s, 6H), 3.81 (s, 3H), 4.52 (t, 2H, J=6.8 Hz), 6.84-7.69 (m, 6H). MS m/e 368 (M.sup.+)

[0151] 46: Yield: 32%. M.p.: 202-203. .sup.1HNMR (DMSO-d6): 2.70-2.84 (m, 8H), 3.80 (s, 6H), 3.90 (s, 3H), 3.95 (s, 3H), 4.39 (t, 2H, J=6.8 Hz), 6.85-7.79 (m, 61-1). MS m/e 364 (M.sup.+)

[0152] 63: Yield: 63%. M.p.: 207-208. .sup.1HNMR (DMSO-d6): 2.67-2.82 (m, 8H), 3.57 (s, 3H), 3.62 (s, 3H), 3.79 (s, 3H), 4.48 (t, 2H, J=7.2 Hz), 6.80-7.64 (m, 6H). MS m/e 352 (M.sup.+)

[0153] 72: Yield: 91%. M.p.: 234-235. .sup.1HNMR (DMSO-d6): 2.70-2.84 (m, 8H), 3.80 (s, 6H), 3.95 (s, 3H), 4.35 (t, 2H, J=6.8 Hz), 6.98-8.44 (m, 6H). MS m/e 413 (M.sup.+)

Example 7

[0154] ##STR00125##

General Method for N-alkylation of 5,6-dihydrobenzo[a]carbazoles Xa. (48, 55, 68, 73)

[0155] 48% HBr (0.5 ml) is added to a solution of N-alkylated carbazole 43, 46, 63, 72 (0.25 mmols) in glacial acetic acid (0.07 ml). The mixture is maintained at reflux and under stirring at 120 C. for 48 hours. After said time, the solvent is evaporated at 1/p, and carbazoles 48 and 73 are recovered pure, directly from the residue obtained. Conversely, for 68 and 55, purification by crystallisation and precipitation of their hydrochlorides is required. MeOH is added to the crude residue, the mixture is cooled in an ice bath, and Et.sub.2OxHCl and anh. Et.sub.2O are added. The resulting precipitate consists of pure hydrochlorides of 55 and 68 which are recovered by vacuum filtration.

[0156] 48: Yield: 67%. M.p.: 228-229. .sup.1HNMR (DMSO-d6): 2.63-2.80 (m, 8H), 3.40 (s, 6H), 4.86 (t, 2H, J=8 Hz), 7.29-8.25 (m, 6H).

[0157] MS m/e 354 (M.sup.+)

[0158] 55: Yield: 86%. M.p.: 139-140. .sup.1HNMR (DMSO-d6): 2.74-2.80 (m, 8H), 3.35 (s, 6H), 4.87 (t, 2H, J=8 Hz), 6.69-8.44 (m, 6H). MS m/e 336 (M.sup.+)

[0159] 68: Yield: 53%. M.p.>330. .sup.1HNMR (DMSO-d6): 2.68-2.79 (m, 8H), 3.39 (s, 6H), 4.70 (t, 2H, J=8 Hz), 6.55-7.11 (m, 6H). MS m/e 337 (M.sup.+)

[0160] 73: Yield: 95%. M.p.>330. .sup.1HNMR (DMSO-d6): 2.75-2.90 (m, 8H), 3.14 (s, 6H), 4.90 (t, 2H, J=8 Hz), 7.10-8.43 (m, 6H).

General Synthesis Method for Acids of Type XIa

Synthesis of 11-(3-dimethylamino-propyl)-8-chloro-5,11-dihydro-6H-benzo[a]carbazol-3-yloxy]-acetic acid 58 and 11-(3-dimethylamino-propyl)-8-fluoro-5,11-dihydro-6H-benzo[a]carbazol-3-yloxy]-acetic acid 70

[0161] K.sub.2CO.sub.3 (23 mmols) is added to a solution of the appropriate 5,6-dihydrobenzo[a]carbazole intermediate Xa (in example 48 or 68) (5.7 mmols) in DMSO (10 ml) until complete solubilisation. A solution of BrCH.sub.2COOH (5.7 mmols) in DMSO (3 ml) is then dripped in at 18 C. After 24 hours, H.sub.2O, ice and CHCl.sub.3 are added to the reaction mixture. The solution is acidified to pH 3, and the resulting precipitate is vacuum filtered. The filtrate is taken up several times with Et0H and dried each time, until the desired acids 58 and 70 are obtained.

[0162] 58: Yield: 47%. M.p.: 189-190 C. .sup.1HNMR (DMSO-d6): 3.14-3.23 (m, 8H), 3.61 (s, 6H), 3.94 (s, 2H), 4.86 (t, 2H, J=8 Hz), 7.02-8.21 (m, 6H). MS m/e 412 (M.sup.+)

[0163] 70: Yield: 14%. M.p.: 22 C. .sup.1HNMR (DMSO-d6): 3.23-3.33 (m, 8H), 3.72 (s, 6H), 3.97 (s, 2H), 4.86 (t, 2H, J=8 Hz), 6.56-8.67 (m, 6H).

Example 8

[0164] ##STR00126##

[0165] General method of quaternisation of amino groups in R.sub.3 of 5,6-dihydrobenzo[a]carbazoles of general formula X-XII

Synthesis of [3-(8-bromo-3-methoxy-5,6-dihydro-benzo[a]carbazol-11-yl)-propyl]-trimethyl-ammonium iodide 82

[0166] A suspension of the appropriate 5,6-dihydrobenzo[a]carbazole intermediate of general formula X-XII (in the example 5,6-dihydrobenzo[a]carbazole 71 xHCl) in saturated solution of NaHCO.sub.3 is extracted several times with AcOEt. The separated organic solvent is dried with MgSO.sub.4 and evaporated at lip to obtain a semisolid residue consisting of 71 free base.

[0167] CH.sub.3I (53 mmols) is added to a solution of 71 free base (7.5 mmols) in anhydrous benzene (76 ml), and the mixture is left under stirring and at reflux for 12 hours. Under these conditions an insoluble solid consisting of the desired quaternary ammonium iodide 82 separates in the reaction solvent, which is vacuum filtered.

[0168] 82: Yield: 77%. M.p.: 223-224. .sup.1HNMR (DMSO-d6): 2.74-2.90 (m, 8H), 3.81 (s, 9H), 3.94 (s, 3H), 4.47 (t, 3H, J=7.2 Hz),

[0169] 6.89-8.56 (m, 6H)

Example 9

[0170] ##STR00127##

Synthesis of 6-hydroxy-tetralone (3)

[0171] A 48% solution of hydrobromic acid (103 mL) is added to a solution of 6-methoxy-1-tetralone (10.0 g, 56.8 mmols) in glacial acetic acid (15 mL). The mixture is maintained at reflux at 120 C. and under stirring for 12 hours. After said time the solvent is evaporated at 1/p to obtain a crystalline solid essentially consisting of 3.

[0172] 3: Yield: 98%. .sup.1HNMR (DMSO-d6): 1.98-3.20 (3m, 6H), 6.5-7.6 and 7.8-8.1 (2m, 3H)

[0173] Similarly to the preparation of 4, acid 30 can be directly obtained by following the method reported below:

Synthesis of 2-(1,2,3,4-tetrahydro-1-oxonaphthalen-7-yloxy) acetic acid (30)

[0174] Potassium carbonate (141.8 mmols) and alpha-bromoacetic acid (49.1 mmols) are added in succession to a solution of 6-hydroxy-1-tetralone 3 (32.7 mmols) in anhydrous acetone (215 mL). The reaction mixture is maintained at reflux and under stirring at 60 C. for 20 hours, after which it is evaporated at 1/p and the solid residue is taken up with H.sub.20 and acidified to pH 4-5 with 10% HCl. The resulting mixture is then extracted 3 times with AcOEt, and the combined organic phases are dried on MgSO.sub.4 and evaporated at 1/p to give the desired acid 30.

[0175] 30: Yield: 63%. .sup.1HNMR (DMSO-d6): 1.97-2.06 (m, 2H), 2.49-2.56 (m, 2H), 2.86-2.92 (t, 2H), 4.77 (s, 2H), 6.85-6.90 (x, 2H), 7.79-7.83 (d, 1H).

Preparation Method of the Hydrazone Intermediates 31 a, b

[0176] The appropriate R-substituted phenylhydrazine (10 mmols) is added to a solution of the appropriate tetralone 4 or 30 (8 mmols) in absolute ethanol (100 mL). The mixture is maintained at room temperature under stirring for 10 hours. After said time the solvent is removed by evaporation at low pressure, and the resulting solid is crystallised from chloroform/hexane to give, depending on the phenylhydrazine used, the desired hydrazones 23a-c or 31a, b.

[0177] 31a: Yield: 47%. .sup.1HNMR (DMSO-d6) : 1.22 (t, 0.1=7.1 Hz, 3H), 1.82-1.87 (m, 2H), 2.57-2.69 (m, 4H), 4.17 (q, 0.1=7.1 Hz, 2H), 4.77 (s, 2H), 6.71 (d, J=2.6 Hz, 1H), 6.80 (dd, J=8.7, 2.6 Hz, 1H), 7.22 (s, 4H), 7.96 (d, J=8.7 Hz, 1H), 10.16 (bs, 1H).

[0178] 31b: Yield: 53%. .sup.1HNMR (DMSO-d6) : 1.22 (t, 0.1=7.1 Hz, 3H), 1.83-1.88 (m, 2H), 2.56-2.72 (m, 4H), 4.17 (q, 0.1=7.1 Hz, 2H), 4.77 (s, 2H), 6.71 (d, J=2.5 Hz, 1H), 5.80 (dd, J=8.7, 2.5 Hz, 1H), 7.16 (d, J=8.8 Hz, 2H), 7.34 (d, J=8.8 Hz, 2H), 7.96 (d, J=8.7 Hz, 1H), 9.27 (bs, 1H).

Method of Preparation of Acid Hydrazones 23a, b

[0179] 3.3 mL of a 1N aqueous solution of NaOH (3.3 mmols) is added to a solution of the appropriate ethyl ester (31a or 31b) (1.9 mmols) in ethanol (1.2 mL). The resulting mixture is then heated to reflux for 1 h. The reaction mixture is cooled to room temperature, poured into 0.4 mL of concentrated HCl (3.6 mmols) and cooled in ice, and the solid that precipitates is collected by filtration and dried under vacuum.

[0180] 23a: Yield: 63%. M.p.: 212-213 C. .sup.1HNMR (DMSO-d6) : 1.81-1.85 (m, 2H), 2.55-2.68 (m, 4H), 4.45 (s, 2H), 6.62 (d, J=2.2 Hz, 1H), 6.75 (dd, J=8.6, 2.2 Hz, 1H), 7.21 (s, 4H), 7.93 (d, J=8.6 Hz, 1H), 9.23 (bs, 1H).

[0181] 23b: Yield: 49%. M.p.>330 C. .sup.1HNMR (DMSO-d6) : 1.81-1.85 (m, 2H), 2.55-2.67 (m, 4H), 4.42 (s, 2H), 3.62 (d, J=1.7 Hz, 1H), 6.74 (dd, J=8.8, 1.7 Hz, 1H), 7.15 (d, J=8.2 Hz, 2H), 7.32 (d, J=8.2 Hz, 2H), 7.92 (d, J=8.8 Hz, 1H), 9.24 (bs, 1H).

Direct Synthesis from Tetralone 30 of Acid Hydrazone 23c

[0182] 4-trifluoromethyl)phenylhydrazine (0.95 g) is added to a solution of acid 30 (0.10 g, 0.45 mmols) in absolute ethanol (30 mL). The mixture is maintained at room temperature under stirring for 3 days. After said time the solvent is removed by evaporation at 1/p, and the resulting solid is crystallised from chloroform/hexane to give 0.091 g of the desired acid 23c.

[0183] 23c: Yield: 53%. M.p.: 196-197 C.: .sup.1H NMR (DMSO-d6) : 1.86 (m, 2H), 2.64-2.72 (m, 4H), 4.69 (s, 2H), 6.72-1H), 6.81 (d, J=8.9 Hz, 1H), 7.34 (d, J=8.4 Hz, 2H), 7.52 (d, J=8.4 Hz,), 8.00 (d, J=8.9 Hz, 1H), 9.60 (bs, 1H).

Example 10

[0184] ##STR00128##

[0185] Reagents and conditions. i: p-Cl-Benzyloxyamine hydrochloride, CH.sub.3CN r/t, 6 days.

Synthesis of [5-(4-chloro-benzyloximino)-5,6,7,8-tetrahydro-naphthalen-2-yloxy]-acetic acid (oxime ether 24)

[0186] A solution of tetralone 30 (0.44 g, 2 mmols) and of p-chloro-benzylhydroxy-amine (0.39 g, 2 mmols) in CH.sub.3CN (60 mL) is maintained under stirring at room temperature for 6 days. After said time, the solvent is evaporated at 1/p and the solid crude product obtained is purified by silica-gel chromatography, eluting with a hexane/AcOEt 1:1 mixture containing 2% AcOH. From the main fractions collected by eliminating the solvent mixture, a solid crystallises which essentially consists of the desired oxime ether 24.

[0187] 24: Yield: 45%. M.p.: 143-144 C.: .sup.1HNMR (CDCl.sub.3) : 0.82 (m, 2H), 2.74 (m, 4H), 4.67 (brs, H exchange=_), 5.15 (s, 2H), 6.60-6.80 (m, 2H), 7.20-7.40 (m, 4H), 7.90 (d, J=8.6 Hz, 1H).

Example 11

[0188] Alternative methods a) and b) for preparation of the following acids: 8-R-(5,11-dihydro-6H-benzo[a]carbazol-3-yloxy)-acetic acids 25a-c of type XIIIa and 8-R-(11H-benzo[a]carbazol-3-yloxy)-acetic acids of type XIIIb and their corresponding ethyl esters 32a, b (type XIIIa) and 33a, b (type XIIIb)

##STR00129##

[0189] Method a)

Synthesis of ethyl esters 32a, b (type XIIIa) and 33a, b (type XIIIb)

[0190] The appropriate 4-R-phenylhydrazine (8.4 moles) is added to a solution of tetralone 29 (2.00 g, 8.05 mmols) in glacial acetic acid (11 mL), and the mixture is maintained at reflux under stirring for 20 hours. After said time the solvent is evaporated at 1/p, and the resulting solid is dissolved in the minimum quantity of Et.sub.20 and washed several times with H.sub.20 and a saturated solution of NaHCO.sub.3. The ether solution is then evaporated at 1/p, supplying the desired ethyl esters (32a,b) and (33a,b) in admixture. .sup.1H-NMR analysis reveals a % composition in admixture of the two carbazoles of type XIIIa (32a) and XIIIb (33a) at the ratio of 84/16, and that of the admixture of 32b (type XIIIa) and 33b (type XIIIb) at the ratio of 80/20.

[0191] 32a: .sup.1HNMR (DMSO-d6) : 1.22 (t, 0.1=7.1 Hz, 3H), 2.90 (m, 4H), 4.18 (q, 0.1=7.1-Hz, 2H), 4.81 (s, 2H), 6.80-7.15 (m, 3H), 7.25-8.62 (m, 3H), 11.55 (s, 1H).

[0192] 33a: .sup.1HNMR (DMSO-d6) : 1.22 (t, 0.1=7.1 Hz, 3H), 4.18 (q, 0.1=7.1 Hz, 2H), 0.90 (s, 2H), 8.15-8.45 (m, 3H), 12.23 (s, 1H).

[0193] 32: .sup.1HNMR (DMSO-d6) : 1.22 (t, 0.1=7.1 Hz, 3H), 2.89 (m, 4H), 4.18 (q, . . . ; =7.1 Hz, 2H), 4.80 (s, 2H), 6.81-7.63 (m, 6H), 11.55 (s, 1H).

[0194] 33b: .sup.1HNMR (DMSO-d6) : 1.23 (t, 0.1=7.1 HZ, 3H), 4.18 (q, 0.1=7.1 Hz, 2H), 0.49 (s, 2H), 7.10-7.60 and 8.10-8.50 (m, 8H), 12.28 (s, 1H).

Acid Synthesis: 8-R-(5,11-dihydro-6H-benzo[a]carbazol-3-yloxy)-acetic acids 25a-c of type XIIIa and 8-R-(11H-benzo[a]carbazol-3-yloxy)-acetic acids of type XIIIb

[0195] KOH (3.6 mmols) is added to a solution of the appropriate ester 32a, b (3 mmols) and absolute EtOH (108 mL), and the mixture is left under stirring at r/t for 20 hours. After said time, the solvent is evaporated at 1/p. The resulting solid is dissolved in the minimum quantity of H.sub.2O and several washes are performed with AcOEt, after which a 10% HCl solution is added until pH 4-5 is reached. The result is a solid which is vacuum filtered. Said solid is then crystallised from EtOH to obtain the acids of type XIIIa 25a and 25b still about 10% contaminated with the corresponding aromatic carbazoles type XIIIb (26a, b).

[0196] 25a: Yield: 12% .sup.1HNMR (DMSO-d6) : 2.89 (m, 4H), 4.70 (s, 2H), 6.80-7.07 (m, 3H), 7.25-0.75 (m, 3H), 11.54 (s, 1H).

[0197] 25b: Yield: 10% .sup.1HNMR (DMSO-d6) : 2.86 (m, 4H), 4.84 (s, 2H), 6.70-6.90 (m, 2H), 7.05-0.40 (m, 2H), 7.50-7.70 (m, 2H), 11.63 (s, 1H).

[0198] Method b)

Acid synthesis: 8-R-(5,11-dihydro-6H-benzo[a]carbazol-3-yloxy)-acetic acids 25a-c

[0199] A solution of tetralone 30 (1.4 mmols) and the appropriate substituted 4-R-phenylhydrazine (1.5 mmols) in glacial acetic acid (2 mL), previously degassed with a stream of Ar, is heated to reflux overnight under an inert atmosphere of Ar. The mixture is then concentrated at 1/p and the residue is purified by flash chromatography on silica gel (eluent hexane/ethyl acetate 1:1+2% acetic acid). The resulting solid is crystallised from EtOH to give practically pure acids of type XIII a (25a-c).

[0200] 25a: Yield: 83%. M.p.: 146-148 C.: .sup.1HNMR (DMSO-d6) : 2.89 (m, 4H), 4.70 (s, 2H), 6.80-7.07 (m, 3H), 7.25-0.75 (m, 3H), 11.54 (s, 1H).

[0201] 25b: Yield: 53%. M.p.: 153-155 C.: .sup.1HNMR (DMSO-d6) : 2.86 (m, 4H), 4.84 (s, 2H), 6.70-6.90 (m, 2H), 7.05-0.40 (m, 2H), 7.50-7.70 (m, 2H), 11.63 (s, 1H).

[0202] 25c: Yield: 63%. M.p.: 243-245 C.: .sup.1HNMR (DMSO-d6) : 2.92-2.98 (m, 4H), 4.71 (s, 2H), 6.91 (dd, J=8.4, 2.5 Hz, 1H), 6.93 (d, J=2.5 Hz, 1H), 7.33 (d, J=8.6 Hz, 1H), 7.52 (d, J=3.6 Hz, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.84 (s, 1H), 11.84 (s, 1H).

Example 12Methods of Aromatising the C Ring of Nuclei Type Xa, XIa, XIIIa, XIVa and XVa

[0203] This type of oxidative conversion typical of 5,11-dihydro-6H-benzo[a]carbazoles of type Xa, XIa, XIIIa, XIVa and XVa to 11H-benzo[a]carbazoles of type Xb, XIb, XIIIb, XIVb and XVb can be obtained as required by methods a)-c) reported below by way of example for oxidation to the 11H-benzo[a]carbazoles 26a, b and 37 of 8-(Cl)-5,6-dihydrobenzo[a]carbazoles 9a and 36 or 8-(Br)-5,6-dihydrobenzo[a]carbazole 9b.

##STR00130##

[0204] Reagents and conditions. i: AcOH, 120 C., 5 days; ii: AcOH, PdC, 120 C., 20 h iii: HBF.sub.4 50%, 0 C., 10 min; iv: CH.sub.3CN, TEMPO, 0 C., 15 min.

[0205] Method a)

Synthesis of (8-bromo-11H-benzo[a]carbazol-3-yloxy)-acetic acid 26a and (8-bromo-11H-benzo[a]carbazol-3-yloxy)-acetic acid 26b

[0206] A solution of the appropriate carbazole (25a, b) (3 mmols) in glacial acetic acid (20 mL) is left at reflux in air for 5 days. After said time the solution is concentrated and the precipitated solid collected by vacuum filtration. The precipitate consisting of the desired products of oxidation (26a, b) is washed with small aliquots of CHCl.sub.3.

[0207] 26a: Yield: 73%. M.p.: 250-253 C. .sup.1H NMR (DMSO-d6) : 4.81 (s; 2H), 7.28-7.47 (m, 3H), 7.50-7.62 (m, 2H), 8.14-8.22 (m, 1H), 8.39-8.44 (d, 1H).

[0208] 26b: Yield: 66%. M.p.: 228-231 C. .sup.1H NMR (DMSO-d6) : 4.39 (s, 2H), 6.76 (x, 1H), 7.20-7.7 (m, 5H), 8.14-8.6 (m, 3H), 12.25 (s, 1H).

[0209] Method b)

Preparation of 2,2,6,6-tetramethylpiperidin-1-oxonium tetrafluoroborate (35)

[0210] A 48% solution of tetrafluoroboric acid (9 mL, 0.06 mmols) is added drop by drop to a solution of TEMPO (2,2,6,6-tetramethyl-1-oxy-piperidine) (34) (5 g, 32.00 mmols) in Et.sub.20 (20 mL). During dripping, the solution is maintained under stirring at the temperature of 0 C. At the end of the addition, the temperature is maintained at 16 C. for 10 minutes. The yellow precipitate that forms during this time, which is collected by filtration, washed with cold ether and dried, consists solely of 35 (3.3 g).

[0211] 35: Yield 41%. M.p.: 164-165 C.

Synthesis of 8-chloro-3-methoxy-11H-benzo[a]carbazole (37)

[0212] A solution of TEMPO (0.17 g, 0.66 mmols) in acetonitrile (1.20 mL) is added by slow dripping to a solution of 8-chloro-3-methoxy-5,11-dihydro-6H-benzo[a]carbazole 36 (0.20 g, 0.70 mmols) in anhydrous acetonitrile (7 mL). The solution is maintained under stirring in an ice bath for 15 minutes, after which the resulting solid is collected by vacuum filtration.

[0213] 37: Yield 99%. M.p.: 165-167 C. .sup.1HNMR (DMSO-d6) : 3.91 (s, 3H), 7.29-7.58 (m, 5H), 8.15-8.43 (m, 3H), 2.25 (s, 1H).

Example 13Example of Reduction of an NO.SUB.2 .Group in R.SUB.1 .on the a Ring of Compounds of Type I-XIII (where Applicable)

[0214] Reduction of 3-methoxy-9-nitro-5,11-dihydro-6H-benzo[a]carbazole 40 to the amine 3-methoxy-11H-benzo[a]carbazol-9-yl 41

[0215] Ni-Raney (66 mg) is added as catalyst under inert atmosphere (Ar) to a solution of 40 (2.00 mmols) in absolute EtOH (37 mL) plus 64% hydrazine hydrate (5 mL). The mixture is left at reflux, under stirring, hydrazine (9.3 mmols) is added drop by drop, and the reflux continues for a further 2 hours. After said time the mixture is filtered through celite under an inert atmosphere (Ar). The filtrate is evaporated at lip, and the solid residue essentially consists of the desired product 41.

[0216] 41: Yield 94%. M.p.: 182-184. .sup.1H NMR (DMSO-d6) : 2.70-3.88 (m, 4H), 4.76 (s, 2H), 6.34-6.54 (m, 2H), 6.77-6.84 (m, 2H), 7.07-7.11 (d, 1H), 7.38-7.42 (d, 1H), 10.67 (s, 1H).

##STR00131##

Reagents and Conditions. i: N.SUB.2.H.SUB.4.H.SUB.2.O, M-Raney, Abs. EtOH, 80 C., 1.5 l

Example 14Example of Use of TEMPO 35 in the Oxidation of a Nucleus of Type 5,6,7,12-tetrahydro-benzo[6,7]cyclohepta[1,2-b]indole (type Xa, XIa, XIIIa, XIVa, XVa)

Example of oxidation of 3,4-dimethoxy-10-nitro-5,6,7,12-tetrahydro-benzo[6,7]cyclohepta[1,2-b]indole 42 to 3,4-dimethoxy-10-nitro-5,12-dihydro-6H-benzo[6,7]cyclohepta[1,2-b]indol-7-one 43

[0217] ##STR00132##

Reagents and Conditions: TEMPO (35), THF, r/t, 6 Days

Synthesis of 3,4-dimethoxy-10-nitro-5,12-dihydro-6H-benzo[6,7]cyclohepta[1,2-b]indol-7-one 43

[0218] A solution of TEMPO (35) (0.23 g, 0.89 mmol) in anh. acetonitrile (2 mL) is added by slow dripping to a solution of 3,4-dimethoxy-10-nitro-5,6,7,12-tetrahydro-benzo[6,7]cyclohepta[1,2-b]indole 42 [obtained as above for the analogues of type Xa as pure XIa, namely by reacting an appropriate R.sub.2-substituted 6,7,8,9-tetrahydro-benzocyclohepten-5-one with the appropriate phenylhydrazine, scheme Id, same conditions for Xa and XIa] (0.3 g, 0.9 mmol) in anhydrous acetonitrile (50 mL), under stirring at 0 C. in an inert atmosphere. The solution is stirred at r/t for 6 days, the solvent is removed by evaporation at lip, and the solid residue obtained is purified by flash chromatography on silica gel (hexane/ethyl acetate 1:1), supplying the desired pure ketone 43 as a glassy yellow oil.

[0219] 43: Yield 28%. .sup.1H NMR (DMSO-d6) : 2.49 (s, 3H), 2.50 (s, 3H), 3.75-3.90 (m, 4H), 7.50-8.50 (m, 5H), 10.1 (s, 1H).

Example 15Biopharmacological Activity Tests

[0220] Measurements of biological activity conducted by the following methods:

[0221] 1) Costa, B; Grillone, A F; Salvetti, A; Rocchiccioli, S; Iacopetti, P; Daniele, S; Da Pozzo, E; Campiglia, P; Novellino, E; Martini, C; Rossi, L An antibody-free strategy for screening putative HDM2 inhibitors using crude bacterial lysates expressing GST-HDM2 recombinant protein Drug Testing and Analysis (2013), 5, 7, 596-601.

[0222] 2) Costa B, Bendinelli S, Gabelloni P, Da Pozzo E, Daniele S, et al. Human Glioblastoma Multiforme: p53 Reactivation by a Novel MDM2 Inhibitor. PLoS ONE (2013) 8(8): e72281. doi:10.1371/journal.pone.0072281.

[0223] Results:

[0224] 1) Inhibitory effect on p53/MDM2 complex binding (IC50 nM/% inhibition at 10 uM) of some ligands of general formula I and of nutlin (Nut-3 used as reference). Test performed on cell lysate of human glioblastoma (GBM) cells U343MG (1,2). The results are set out in Table 2.

TABLE-US-00002 TABLE 2 # # substructure Code Structure IC50 nM Comparator Medicament Nut-3 [00133] 296 15 XIc XIl RA1 [00134] 920 20 XIa RA2 [00135] 2900 120 XIr RA3 [00136] 620 50 Xa GA17M8 [00137] 12700 1500 XIb RM66 [00138] 2700 120 XIb RM58 [00139] (58% a 10 uM) XIb RM85 [00140] (82% a 10 uM) XI'd RA6 [00141] (18% a 10 uM) XI'd RA7 [00142] n.a. XI'd RA9 [00143] n.a. XI'd RA11 [00144] n.a. XI'd RA12 [00145] n.a. XI'd RA13 [00146] (22% a 10 uM) XI'g RA14 [00147] n.a. X'c RA15 [00148] (<5% a 10 uM) X'c RA16 [00149] (<5% a 10 uM) XI'c RA17 [00150] n.a. XI'c RA18 [00151] 1350 150 XI'c RA19 [00152] 450 15 XI'h RA20 [00153] 1550 250 XIV'a GA09a [00154] 650 50 XIV'a' GA09a' [00155] n.a. XIV'a GA09b [00156] 225 20 XIV'a' GA09b' [00157] n.a. XIV'a RM36 [00158] 280 20 XIV'a RM37 [00159] 222 44 XIV'a' RM53 [00160] (<5% a 10 uM) XIV'a RM47 [00161] 195 22 XIV'd RA25 [00162] 143 16 XV'e RA26 [00163] 120 90 XV'g' RA29 [00164] 100 30 XIIId RA30 [00165] 150 10 XIV'c RA31 [00166] 120 45 XV'c RA35 [00167] 375 22 XIV'e RA36 [00168] 220 100 XIV'g' RA37 [00169] 180 150 XIV'g' RM38 [00170] 222 210 XIV'g RM39 [00171] 232 200 XIVr RA40 [00172] 197 124 XVr RA42 [00173] 292 25 XIVc RM43 [00174] 228 16 XIIIr RA44 [00175] 223 22

[0225] 2) Stabilisation of p53 (U343MG, 1, 2).

[0226] The stabilisation of p53 in the presence of RM37 or Nut-3 is due to the reduction of p53 bonded to MDM2, not induction from scratch of mRNA synthesis of p53 (1,2). The results are set out in FIG. 1.

[0227] 3) Effect of RM37 and Nut-3 on activation of the gene transactivation function of p53 (U343MG; 1,2). The results are set out in FIG. 2.

[0228] 4) RM37 stabilises the intracellular levels of protein p53 (343MG; 1,2). The results are set out in FIG. 3.

[0229] 5) Antitumoral effect in vitro (RM37 vs Nut-3): dead cell count (U343MG; 1,2). The results are set out in FIG. 4.

[0230] 6) Antitumoral effect in vitro (RM37 vs Nut-3): live cell count (U343MG; 1,2). The results are set out in FIG. 5.

[0231] 7) Effect of RM37 on cell cycle (U343MG; 1,2). RM37 blocks the cell cycle in G1. The results are set out in FIG. 6.

[0232] 8) Effect of RM37 on cell apoptosis (U343MG; 1,2). RM37 induces cell apoptosis (U343MG). The results are set out in FIG. 7 and FIG. 8.